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	<front>
		<journal-meta>
			<journal-id journal-id-type="publisher-id">rmfi</journal-id>
			<journal-title-group>
				<journal-title>Revista mexicana de fitopatología</journal-title>
				<abbrev-journal-title abbrev-type="publisher">Rev. mex. fitopatol</abbrev-journal-title>
			</journal-title-group>
			<issn pub-type="ppub">0185-3309</issn>
			<issn pub-type="epub">2007-8080</issn>
			<publisher>
				<publisher-name>Sociedad Mexicana de Fitopatología A.C.</publisher-name>
			</publisher>
		</journal-meta>
		<article-meta>
			<article-id pub-id-type="doi">10.18781/R.MEX.FIT.2024-23</article-id>
			<article-categories>
				<subj-group subj-group-type="heading">
					<subject>Articles</subject>
				</subj-group>
			</article-categories>
			<title-group>
				<article-title>Biological effectiveness of <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts against <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani in vitro</italic></article-title>
			</title-group>
			<contrib-group>
				<contrib contrib-type="author">
					<name>
						<surname>Ramírez-Méndez</surname>
						<given-names>Jesús Eduardo</given-names>
					</name>
					<xref ref-type="aff" rid="aff1">¹</xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Hernández-Castillo</surname>
						<given-names>Francisco Daniel</given-names>
					</name>
					<xref ref-type="aff" rid="aff2">²</xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Gallegos-Morales</surname>
						<given-names>Gabriel</given-names>
					</name>
					<xref ref-type="aff" rid="aff3">³</xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Jasso-Cantú</surname>
						<given-names>Diana</given-names>
					</name>
					<xref ref-type="aff" rid="aff4">⁴</xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Arredondo-Valdés</surname>
						<given-names>Roberto</given-names>
					</name>
					<xref ref-type="aff" rid="aff5">⁵</xref>
				</contrib>
				<contrib contrib-type="author">
					<name>
						<surname>Tucuch-Pérez</surname>
						<given-names>Marco Antonio</given-names>
					</name>
					<xref ref-type="aff" rid="aff6">⁶</xref>
				</contrib>
			</contrib-group>
			<aff id="aff1">
				<institution content-type="original">¹ Departamento de Parasitología, Universidad Autónoma Agraria Antonio Narro, Calzada Antonio Narro 1923, Buenavista, CP. 25315, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma Agraria Antonio Narro</institution>
				<institution content-type="orgname">Universidad Autónoma Agraria Antonio Narro</institution>
				<addr-line>
					<postal-code>25315</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<aff id="aff2">
				<institution content-type="original">² Departamento de Parasitología, Universidad Autónoma Agraria Antonio Narro, Calzada Antonio Narro 1923, Buenavista, CP. 25315, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma Agraria Antonio Narro</institution>
				<institution content-type="orgname">Universidad Autónoma Agraria Antonio Narro</institution>
				<addr-line>
					<postal-code>25315</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<aff id="aff3">
				<institution content-type="original">³ Departamento de Parasitología, Universidad Autónoma Agraria Antonio Narro, Calzada Antonio Narro 1923, Buenavista, CP. 25315, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma Agraria Antonio Narro</institution>
				<institution content-type="orgname">Universidad Autónoma Agraria Antonio Narro</institution>
				<addr-line>
					<postal-code>25315</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<aff id="aff4">
				<institution content-type="original">⁴ Departamento de Parasitología, Universidad Autónoma Agraria Antonio Narro, Calzada Antonio Narro 1923, Buenavista, CP. 25315, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma Agraria Antonio Narro</institution>
				<institution content-type="orgname">Universidad Autónoma Agraria Antonio Narro</institution>
				<addr-line>
					<postal-code>25315</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<aff id="aff5">
				<institution content-type="original">⁵ Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Ing. J. Cárdenas Valdez S/N, Col. República, CP. 25280, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma de Coahuila</institution>
				<institution content-type="orgname">Universidad Autónoma de Coahuila</institution>
				<addr-line>
					<postal-code>25280</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<aff id="aff6">
				<institution content-type="original">⁶ Facultad de Ciencias Químicas, Universidad Autónoma de Coahuila, Ing. J. Cárdenas Valdez S/N, Col. República, CP. 25280, Saltillo, Coahuila, México.</institution>
				<institution content-type="normalized">Universidad Autónoma de Coahuila</institution>
				<institution content-type="orgname">Universidad Autónoma de Coahuila</institution>
				<addr-line>
					<postal-code>25280</postal-code>
					<city>Saltillo</city>
					<state>Coahuila</state>
				</addr-line>
				<country country="MX">Mexico</country>
			</aff>
			<pub-date date-type="pub" publication-format="electronic">
				<day>23</day>
				<month>11</month>
				<year>2024</year>
			</pub-date>
			<pub-date date-type="collection" publication-format="electronic">
				<year>2024</year>
			</pub-date>
			<volume>42</volume>
			<issue>4</issue>
			<elocation-id>RMEXFIT202423</elocation-id>
			<history>
				<date date-type="received">
					<day>02</day>
					<month>07</month>
					<year>2024</year>
				</date>
				<date date-type="accepted">
					<day>09</day>
					<month>11</month>
					<year>2024</year>
				</date>
			</history>
			<permissions>
				<license license-type="open-access" xlink:href="https://creativecommons.org/licenses/by-nc/4.0/" xml:lang="en">
					<license-p>This is an open-access article distributed under the terms of the Creative Commons Attribution License</license-p>
				</license>
			</permissions>
			<abstract>
				<title>Abstract:</title>
				<sec>
					<title>Background/Objective. </title>
					<p><italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic> are pathogens that affect agricultural crops. The objective of this study was to evaluate some <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> methanolic extracts, from the Chihuahuan Desert, against those fungi, in search for biological control alternatives.</p>
				</sec>
				<sec>
					<title>Materials and Methods. </title>
					<p>Pathogens from chili plants were isolated and identified using morphological and molecular methods. Methanolic extracts from both plants were prepared, and the antioxidant capacity (AC), the total polyphenol content (TPC), and the antifungal compounds via HPLC-MS were assessed. The antifungal effectiveness was tested at concentrations of 3.9-2000 mg L<sup>-1</sup> using a poisoned medium assay, where fungi structural damage was observed.</p>
				</sec>
				<sec>
					<title>Results. </title>
					<p>The <italic>F. splendens</italic> and <italic>A. striata</italic> extracts exhibited 55% and 68% AC, as well as 61 mg/g and 112 mg/g TPC, respectively. Both extracts contained caffeic acid and quercetin, while <italic>F. splendens</italic> also exhibited eriodyctiol, kaempferol, and luteolin; <italic>A. striata</italic> contained pinocembrin and theaflavin B. <italic>F. splendens</italic> attained 100% inhibition of <italic>P. aphanidermatum</italic> at 250 mg L<sup>-1</sup>, and of <italic>R. solani</italic> at 500 mg L<sup>-1</sup>, whereas <italic>A. striata</italic> achieved 100% inhibition at 1000 mg L<sup>-1</sup> in both cases. The extracts produced lysis in <italic>P. aphanidermatum</italic> oogonia and mycelial fragmentation in <italic>R. solani</italic>.</p>
					<p> </p>
				</sec>
				<sec>
					<title>Conclusion. </title>
					<p>The <italic>F. splendens</italic> and <italic>A. striata</italic> methanolic extracts demonstrate</p>
					<p>promising antifungal activity against <italic>P. aphanidermatum</italic> and <italic>R. solani</italic>, suggesting that these natural compounds might be useful as a biological alternative for pathogen</p>
					<p>control in agricultural crops.</p>
				</sec>
			</abstract>
			<kwd-group xml:lang="en">
				<title>Keywords:</title>
				<kwd>plant extracts</kwd>
				<kwd>biological effectiveness</kwd>
				<kwd>phytochemicals</kwd>
				<kwd>phytopathogenic fungi.</kwd>
			</kwd-group>
			<counts>
				<fig-count count="8"/>
				<table-count count="6"/>
				<equation-count count="0"/>
				<ref-count count="46"/>
				<page-count count="0"/>
			</counts>
		</article-meta>
	</front>
	<body>
		<sec sec-type="intro">
			<title>Introduction</title>
			<p><italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic> are widely recognized pathogens due to their ability to cause devastating diseases in a variety of important agricultural crops. <italic>P. aphanidermatum</italic> causes rotting of the root, it affects the root system and causes the death of preemergent seedlings via aqueous lesions in the root and stem (<xref ref-type="bibr" rid="B33">Punja and Yip, 2003</xref>; <xref ref-type="bibr" rid="B15">Grijalba <italic>et al.</italic>, 2015</xref>). On the other hand, <italic>R. solani</italic> affects stems from the base, causing soft rots and root damages that weaken and kill plants prematurely (<xref ref-type="bibr" rid="B29">Medeiros <italic>et al</italic>., 2015</xref>). The persistence and adaptation of these phytopathogens in the soil complicate their management, affecting food security and the agricultural economy. Due to this, sustainable alternatives are currently being sought, such as plant extracts, with plants from the Chihuahuan Desert standing out as an option for the production of plant extracts due to their phytochemical composition and their ability to inhibit the development of phytopathogenic microorganisms (Tucuch-Pérez <italic>et al</italic>., 2021).</p>
			<p>In this sense, <italic>Fouquieria splendens</italic> and <italic>Agave striata</italic> coexist and develop within the Chihuahan Desert. <italic>A. striata</italic> or “espadín” agave has a wide distribution in the south, from the Mexican state of Coahuila down to Hidalgo (<xref ref-type="bibr" rid="B13">Gentry, 1982</xref>), whereas <italic>F. splendens</italic> or “ocotillo” is distributed from southwestern United States to southern Zacatecas, Querétaro and Hidalgo (<xref ref-type="bibr" rid="B16">Henrickson, 1972</xref>). Investigations indicate that plants of the Chihuahuan Desert can inhibit or eliminate phytopathogenic microorganisms (Lira-Saldívar, 2003) that cause devastating diseases in economically important crops (Martínez <italic>et al</italic>., 2016; Hernández <italic>et al</italic>., 2018).</p>
			<p>The control of phytopathogenic microorganisms is attributed to the environmental factors found in these regions, which stimulate the production of antimicrobial compounds in wild plants (Larios <italic>et al</italic>., 2020; Salas <italic>et al</italic>., 2023). The Agavaceae and Fouquieriaceae families have stood out for their antimicrobial properties (Gonzáles <italic>et al</italic>., 2015; Ramírez <italic>et al</italic>., 2023). <italic>A. striata</italic> and <italic>F. splendens</italic> are currently known to be used in the treatment of diverse illnesses in traditional medicine (Pérez <italic>et al</italic>., 2003; López <italic>et al</italic>., 2022), and have displayed antimicrobial properties against <italic>Clavibacter michiganensis</italic> subsp <italic>michiganensis</italic>, presenting phytochemicals such as alkaloids, carbohydrates, flavonoids, saponins, tannins and quinones (Ramírez <italic>et al</italic>., 2023). The aims of this investigation were to identify phytochemical compounds found in methanolic <italic>A. striata</italic> and <italic>F. splendens</italic> extracts, to determine their Antioxidative Capacity (AC) and Total Polyphenol Content (TPC), to evaluate their biological effectiveness <italic>in vitro</italic> on <italic>P. aphanidermatum</italic> and <italic>R. solani,</italic> and to observe morphological changes in their structures due to the plant extracts.</p>
		</sec>
		<sec sec-type="materials|methods">
			<title>Materials and Methods</title>
			<p>Collecting plants. <italic>A. striata</italic> leaves and <italic>F. splendens</italic> stems were gathered in the municipal area of General Cepeda, Coahuila, Mexico (25°20’27.13’N and 101°27’52.07’’W at 1531 m.a.s.l.), in the month of August. They were placed in black plastic bags and transported to the Mycology and Biotechnology Laboratory of the Agricultural Parasitology Department of the Universidad Autónoma Agraria Antonio Narro. The plant material was washed with tap water, then placed in a drying oven (Arsa AR-130D, Mexico) at 60 °C until its weight became constant. Finally, they were ground with a mill (Surtek Mogra1, Mexico) and sieved with a 0.2 mm sieve for storage in dark and properly labelled containers until use.</p>
			<p>Preparation of the extracts. The extracts were prepared following the method reported by Jasso <italic>et al</italic>. (2015), with some modifications, using 96% methanol as a solvent; 42 g of the powder of each plant were deposited in 500 mL flasks adding 375 mL of the solvent, and they were placed in a stirring hotplate (Thermo Scientific Cimarec, USA) for 72 h at 60 °C. The solvent was then separated using rotary evaporation (IKA RV 10 digital V, USA) at 150 rpm at 70 °C. Once the methanolic fraction was obtained, it was poured into glass containers and placed in a drying oven (Arsa AR-130D, Mexico) at 50 °C for 72 h. Finally, the extracts were pulverized and stored in amber colored bottles in refrigeration at 4 °C.</p>
			<p><bold>Characterization of phytochemicals found in the <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts with Reverse-phase liquid chromatography (HPLC-MS).</bold> The reverse-phase High-Performance Liquid Chromatography was carried out following the method by Ascacio <italic>et al</italic>. (2016), which consists of using an HPLC system. It varies in that it includes an automatic injector (Varian ProStar 410, USA), a three-way pump (Varian ProStar 2310, USA) and a PDA decanter (Varian ProStar 330, HPLC-MS). A mass spectrometer with a liquid chromatograph ion trap (Varian 500 - MS IT Mass Spectrometer, USA), equipped with a source of electrons by electrospraying was also used. Samples (5 μL) were injected into a Denali C18 column (150 mm x 2.1 mm, 3 μm, Grace, USA). The oven temperature was maintained at 30 °C. The eluents were formic acid (0.2 %, V/V; solvent A) and acetonitrile (solvent B). The following gradient was applied: initial, 3% B; 0 - 5 min, 9% B linear; 5 - 15 min, 16% B linear; 15 - 45 min, 50% B linear. After the column was washed and reconditioned, the flow rate was maintained at 0.2 mL/ min and the elution was monitored at 245, 280, 320 and 550 nm. All the effluent (0.2 mL/min) was injected into the mass spectrometer source, without splitting. All MS experiments were conducted in a negative mode [M-H]-1. Nitrogen was used as a nebulizer gas and helium, as the buffer gas. The ion source parameters were: spray voltage 5.0 kV and capillary voltage and temperature were 90.0 V and 350°C, respectively. The data were gathered and processes using the MS Workstation Software (V 6.9). The samples were first analyzed in full-scan mode, acquired in the m/z range 50 - 2000. 6,951.6</p>
			<p><bold>Antioxidant capacity DPPH in <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts.</bold> This was determined according to descriptions by Brand <italic>et al</italic>. (1995) with some modifications and analyzed by spectrophotometry. For this, 50 mg of dry material was weighed, which was added 1.0 mL of 80% methanol. Subsequently, the samples were sonicated for 20 min and centrifuged for 15 min at 14000 rpm. Once the extract was obtained, an aliquot of 10 µL was taken and placed in 96-well microplates and 290 µL of DPPH reagent prepared with ethanol at 100 µM was added. The sample was then allowed to react in the dark for 30 min. After this time, the absorbance was measured at 515 nm. The DDPH percentage of inhibition was determined using the natural absorbance of methanol at 80% as a control. The AC DPPH was determined using the following formula:</p>
			<p>Inhibition of DPPH (%) = Absorbance of target /Absorbance of extract × 100</p>
			<p><bold>Total capacity of polyphenols in <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts.</bold> The TPC was determined using the Folin-Ciocalteu method described by Singleton and Rossi (1965) with some modifications. In the analysis, a 10 µL alquot was taken from the methanol extract at 80%, followed by the addition of 790 µL of distilled water, 50 µL of Folin and 150 µL of sodium carbonate at 20%. Subsequently, the sample was left to rest for 1 h in the dark, after which absorbance was measured in a microplate reader at 765 nm. The TPC was determined using a standard curve with gallic acid and expressed as GAE mg/g (equivalent milligrams of gallic acid per gram).</p>
			<p><bold>Isolation and identification of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> Roots and stems of serrano pepper (<italic>Capsicum annuum</italic>) plants, Var. Platino, were gathered in the stage of flowering, with symptoms of wilting, in the municipal area of Escuinapa, Sinaloa, Mexico, with a randomized sampling taking six plants per sample, and five tissue samples were planted for every Petri dish, performing six repetitions. The samples were disinfected following the methodology by <xref ref-type="bibr" rid="B7">Booth (1977</xref>), they were washed, cut and submerged in 3% chlorine for 3 min, to then be rinsed and dried. They were planted in a PDA medium and incubated at 28 °C for seven days. Fungi were purified with the hyphal tip technique, a method that helps obtain axenic or contaminant-free fungi. Subsequently, the morphological identification was performed using the taxonomic keys by Díaz <italic>et al</italic>. (2011) for the characterization of <italic>P. aphanidermatum,</italic> and by <xref ref-type="bibr" rid="B43">Watanabe and Matsuda (1966</xref>) for</p>
			<p><italic>R. solani</italic>. The molecular identification was carried out by the National Laboratory of Agriculrutal, Medical and Environmental Biotechnology (LANBAMA) at the Instituto Potosino de Investigación Científica y Tecnológica (IPICYT), using the ITS1 and ITS4 primers, which amplify the ITS region between the 18S ribosomal subunit, enabling a specific identification of the fungal species.</p>
			<p><bold>Antifungal activity of <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts on the mycelial growth of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> The antifungal activity was determined by the poisoned medium method proposed by Jasso <italic>et al</italic>. (2011). <italic>A. striata</italic> and <italic>F. splendens</italic> extracts were used at concentrations of 3.9 to 2000 mg L<sup>-1</sup>, in addition to a negative control, which consisted of only a PDA culture without extract; four repetitions were used for every concentration. Discs measuring 0.4 mm in diameter with active <italic>P. aphanidermatum</italic> and <italic>R. solani</italic> mycelia and seven days’ growth were placed in the poisoned medium and incubated at 28 ± 2 °C. The evaluation was carried out measuring the radial growth of the fungi on a daily basis, and concluded when the control completely covered the Petri dish; the experiment was held twice. The percentage of inhibition was determined with the following formula:</p>
			<p>Percentage of Inhibition= (DC-DT/DC) *100.</p>
			<p>where DC is the diameter of the control treatment and DT is the diameter of the different concentrations.</p>
			<p><bold>Effect of <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts on the structures and morphology of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> The methodology by <xref ref-type="bibr" rid="B24">Khaledi <italic>et al</italic>. (2015</xref>) was used, with modifications, to evaluate the modifications induced by the extracts in <italic>P. aphanidermatum</italic> oogonia and <italic>R. solani</italic> hyphae, using the results from the 50% inhibitory concentration (CI<sub>50</sub>) of <italic>A. striata</italic> and <italic>F. splendens</italic>. In order to obtain <italic>P. aphanidermatum</italic> oogonia, the poisoned medium was used with the concentrations of 199.17 and 61.22 mg L<sup>-1</sup> with the <italic>A. striata</italic> and <italic>F. splendens</italic> extracts, respectively, in an Agar V8 culture médium (20 g agar, 3 g of CaCO3, 160 mL of Campbell’s V8 vegetable juice and 840 mL of distilled water), while for the growth of the <italic>R. solani</italic> mycelia, the PDA culture medium in the poisoned medium was used with the extracts at concentrations of 211.18 and 92.59 mg L<sup>-1</sup>; in both experiments, a negative control without any extract was used. After seven days of cultivation, the morphological changes were evaluated under an optic microscope with 40X and 100X lenses. Both experiments were held using six repetitions and were conducted twice.</p>
			<p>Statistical analysis. A Probit analysis was conducted of the <italic>in vitro</italic> trials to determine the IC<sub>50</sub> of each concentration using the SAS program, V9.0., and with the results, an analysis of variance was carried out with the inhibitory concentrations; Tukey tests (p&lt;0.05) were also carried out.</p>
		</sec>
		<sec sec-type="results|discussion">
			<title>Results and Discussion</title>
			<p>
				<inline-graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-i002.gif"/>
				<inline-graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-i002.gif"/><bold>Characterization of phytochemicals found in <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts with Reverse-phase liquid chromatography (HPLC-MS).</bold> The phytochemicals identified in the <italic>A. striata</italic> and <italic>F. splendens</italic> extracts are shown in <xref ref-type="table" rid="t1">Table 1</xref>. In the case of <italic>A. striata</italic>, the presence of two elegitannins (pedunculagin and terflavin B) was observed, along with a hydroxycinnamic acid (caffeic acid), a methoxyflavonol (brassidin), a flavonol (quercetin), a flavonone (pinocepmbrin) and a phenolic terpene (rosmadial). Several bioactive compounds have been reported in <italic>A. striata</italic>, including flavanols (derived from quercetin) (Almaraz <italic>et al</italic>., 2013), triterpenoid and steroidal saponins, as well as alkaloids, carbohydrates, flavonoids (flavanones and chalcones), saponins (triterpenoid and steroidal), tannins (phenol), quinones (anthraquinones and benzoquinones) and coumarins (Ramírez <italic>et al</italic>., 2023). In the case of <italic>F. splendens</italic>, the presence of two flavonols (kaempferol and quercetin) was found, along with two flavones (roifolin and leutin), two methoxyflavonols (patuletin and dimethylquercetin), one flavanone (eriodictyol), one hydroxycinnamic acid (caffeic acid), one dihydroflavonol (dihydroquercetin), and another polyphenol (Phlorin). In this sense, Nevárez-Prado <italic>et al</italic>. (2021) mention that the most characteristic phytochemicals of the <italic>Fouquieria</italic> genus correspond to flavonoids (leucocyanidin, kaempferol and quercetin), p-coumaric, caffeic, ferulic and ellagic acids, as well as coumarin and scopoletin. On the other hand, Rodríguez (2010) found carbonyl groups, phenolic hydroxyls, sterols, methylsterols, coumarins, saponins, sesquiterpene lactones and flavonoids in <italic>F. splendens</italic>. In the stems and leaves, flavonoids such as kaempferol, ermanine and cetine have been found, along with flavones such as apigenin, acacetin, luteolin and chrysoeriol. Among these, apigenin, ermanine, rutin and quercitin-3-O-glucoside were the</p>
			<p>
				<table-wrap id="t1">
					<label>Table 1</label>
					<caption>
						<title>Phytochemical compounds identified in methanolic <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts characterized by reverse-phase liquid chromatography (HPLC-MS).</title>
					</caption>
					<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
						<thead>
							<tr>
								<th>Specie</th>
								<th>Phytochemical</th>
								<th>Retention Time (min)</th>
								<th>Mass</th>
								<th>CAS</th>
								<th>Family</th>
							</tr>
						</thead>
						<tbody>
							<tr>
								<td rowspan="7" style="font-style:italic">Agave striata</td>
								<td>Caffeic acid 4-O-glucoside</td>
								<td>5.554</td>
								<td>341.1</td>
								<td>166735-99-5</td>
								<td>Hydroxycinnamic acids</td>
							</tr>
							<tr>
								<td>Pinocembrin</td>
								<td>21.192</td>
								<td>255.0</td>
								<td>480-39-7</td>
								<td>Flavanones</td>
							</tr>
							<tr>
								<td>Pedunculagin II</td>
								<td>30.902</td>
								<td>785.1</td>
								<td>87687-52-3</td>
								<td>Ellagitannins</td>
							</tr>
							<tr>
								<td>Terflavin B</td>
								<td>35.351</td>
								<td>784.9</td>
								<td>103744-86-1</td>
								<td>Ellagitannins</td>
							</tr>
							<tr>
								<td>Brasidine</td>
								<td>40.341</td>
								<td>623.1</td>
								<td>17331-71-4</td>
								<td>Methoxyflavonols</td>
							</tr>
							<tr>
								<td>Quercetin 3-O-glucuronide</td>
								<td>43.630</td>
								<td>477.2</td>
								<td>22688-79-5</td>
								<td>Flavonols</td>
							</tr>
							<tr>
								<td>Rosmadial</td>
								<td>49.237</td>
								<td>343.9</td>
								<td>85514-31-4</td>
								<td>Phenolic terpenes</td>
							</tr>
							<tr>
								<td rowspan="8" style="font-style:italic">Fouquieria splendens</td>
								<td>Kaempferol 3-O-rutinoside</td>
								<td>37.924</td>
								<td>593.1</td>
								<td>17650-84-9</td>
								<td>Flavonols</td>
							</tr>
							<tr>
								<td>3,7-dimethylquercetin</td>
								<td>39.085</td>
								<td>329.0</td>
								<td>2068-02-2</td>
								<td>Methoxyflavonols</td>
							</tr>
							<tr>
								<td>Dihydroquercetin</td>
								<td>40.224</td>
								<td>303.0</td>
								<td>480-18-2</td>
								<td>Dihydroflavonols</td>
							</tr>
							<tr>
								<td>Rhoifolin 4’-O-glucoside</td>
								<td>40.506</td>
								<td>739</td>
								<td>31498-83-6</td>
								<td>Flavones</td>
							</tr>
							<tr>
								<td>Luteolin 7-O-rutinoside</td>
								<td>41.399</td>
								<td>593</td>
								<td>20633-84-5</td>
								<td>Flavones</td>
							</tr>
							<tr>
								<td>Quercetin 3-O-glucuronide</td>
								<td>43.62</td>
								<td>480.2</td>
								<td>22688-79-5</td>
								<td>Flavonols</td>
							</tr>
							<tr>
								<td>Eriodictyol</td>
								<td>43.803</td>
								<td>287.0</td>
								<td>552-58-9</td>
								<td>Flavanones</td>
							</tr>
							<tr>
								<td>Phlorin</td>
								<td>47.363</td>
								<td>286.9</td>
								<td>28217-60-9</td>
								<td>Other polyphenols</td>
							</tr>
						</tbody>
					</table>
				</table-wrap>
			</p>
			<p>main phenolic compounds in the leaves (<xref ref-type="bibr" rid="B45">Wollenweber, 1994</xref>; <xref ref-type="bibr" rid="B30">Monreal-García <italic>et al</italic>., 2019</xref>). Triterpenes, triterpenoids, steroidal saponins and iridoid compounds have also been discovered have also been discovered (<xref ref-type="bibr" rid="B40">Takhtajan, 2009</xref>), along tieh phytosterols and long- and short-chain alkanes in the flowers; and compounds such as Dammaran-triterpene, fouquierol and isofouquierol in the stem, as well as dammarendiol in the roots (<xref ref-type="bibr" rid="B19">Hegnauer, 1989</xref>; <xref ref-type="bibr" rid="B44">Waterman 1985</xref>). On the other hand, pyxinol and ocotillol have been isolated, the latter being a triterpenoid saponin with antimicrobial activities (<xref ref-type="bibr" rid="B4">Bi <italic>et al</italic>., 2017</xref>). López <italic>et al</italic>. (2022) identified the main phenols in <italic>F. splendens</italic> leaves such as gallic acid, ellagic acid and kaempferol-3-β- glucoside. Furthermore, Ramírez <italic>et al</italic>. (2023) reported that the methanolic extract from <italic>F. splendens</italic> stems contains alkaloids, carbohydrates, flavonoids (flavanones, flavones, flavonols and chalcones), reducing sugars, saponins (triterpenoids), tannins (gallic acid derivatied, catechol derivatives, and phenols), and quinones (anthraquinones and benzoquinones). Plants, within their metabolisms, produce compounds with antimicrobial activities that can control diseases caused by phytopathogenic microorganisms. The extraction of these compounds and their analysis allow for their application against various phytopathogens, offering an alternative for the management of diseases in agriculture (<xref ref-type="bibr" rid="B18">Hernández-Lauzardo <italic>et al</italic>., 2007</xref>). In this sens, Ramírez <italic>et al</italic>. (2023) observed that <italic>A. striata</italic> and <italic>F. splendens</italic> extracts had antimicrobial activities against <italic>Clavibacter michiganensis in vitro</italic> and under greenhouse conditions, with the <italic>F. splendens</italic> extract being the most effective at reducing the incidence and severity of the disease. The <italic>A. striata</italic> and <italic>F. splendens</italic> extracts have displayed antifungal activity in further studies (Sánchez <italic>et al</italic>., 2005; <xref ref-type="bibr" rid="B42">Wang <italic>et al</italic>., 2010</xref>). However, their specific antifungal effect on <italic>R. solani</italic> and <italic>P. aphanidermatum</italic> has not been studied.</p>
			<p><bold>Antioxidant capacity DPPH in <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts.</bold> The results showed that the <italic>F. splendens</italic> extract displayed a significantly greater antioxidant activity in comparison with the <italic>A. striata</italic> extract (<xref ref-type="table" rid="t2">Table 2</xref>). The inhibition of the DPPH radical was expressed as a percentage (%), being 68.7 for <italic>F. splendens</italic> and 55.9 for <italic>A. striata</italic>. In this sense, <xref ref-type="bibr" rid="B12">Garza <italic>et al</italic>. (2012</xref>) determined that the <italic>F. splendens</italic> methanolic extract had a mean effective concentration (EC<sub>50</sub> considered as the sample concentration required to trap 50% of free DPPH radicals) of 130.2 μg. López <italic>et al</italic>. (2022) determined the antioxidant activity of <italic>F. splendens</italic> using DPPH results (2430.1 µmol TE/g), a Trolox Equivalent Antioxidant Capacity (TEAC) of 60.4 µmol TE/g, an Oxygenated Radical Absorbance Capacity (ORAC) of 4948.4 µmol TE/g and a Ferric Reducing Ability of Plasma (FRAP) of 7803 µmol Fe(II)/g) (p &lt; 0.0001). In <italic>Agave</italic> there are investigations in the determinations of the antioxidant capacity. A study conducted by Ahumada <italic>et al</italic>. (2013) obtained results that displayed significant variations in the antioxidant activity between different species of <italic>Agave</italic>. In <italic>Aagave rzedowskiana</italic>, it presented the greatest antioxidant activity with 27.4 μM TE/g p.s., followed by <italic>Agave ornithobroma</italic> with 19.9 μM TE/g p.s.</p>
			<p>
				<table-wrap id="t2">
					<label>Table 2</label>
					<caption>
						<title>Antioxidant Capacity and Capacity of Total Polyphenols of methanolic <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts.</title>
					</caption>
					<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
						<thead>
							<tr>
								<th>Specie</th>
								<th>AC Percentage of Inhibition</th>
								<th>CTP (GAE mg/g)</th>
							</tr>
						</thead>
						<tbody>
							<tr>
								<td style="font-style:italic">Agave striata</td>
								<td>55.90 ± 1.63<sup>a</sup></td>
								<td>61.2 ± 1.17<sup>a</sup></td>
							</tr>
							<tr>
								<td style="font-style:italic">Fouquieria splendens</td>
								<td>68.71 ± 1.22<sup>a</sup></td>
								<td>112.2 ± 3.69<sup>b</sup></td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN1">
							<p>*Values with the same letter are statistically similar (Tukey, p&lt;0.05).</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p><bold>Capacity of total polyphenols in <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts.</bold> The results showed that the <italic>F. splendens</italic> extract displayed significantly higher total polyphenols in comparison with the <italic>A. striata</italic> extract. The TPC was expressed in CFT values CFT (GAE mg/g), being 112.2 for <italic>F. splendens</italic> and 61.2 for <italic>A. striata</italic> (<xref ref-type="table" rid="t2">Table 2</xref>). López <italic>et al</italic>. (2022) analyzed the phenolic compounds of the methanolic extract taken from <italic>F. splendens</italic> leaves, using the method by Folin Ciocalteu and after fractioning, they noticed that the ethyl acetate fraction contained the greatest number of phenolic compounds, with 479.9 mg GAE/g (p &lt; 0,0001). In another study, Ahumada <italic>et al</italic>. (2013) found significant variations in the total phenol contents between species of <italic>Agave</italic>. <italic>A. ornithobroma</italic> had the highest value with 12.37 mg GAE/g p.s., while <italic>Agave angustifolia</italic> displayed the lowest value with 2.06 mg GAE/g p.s., indicating a considerable variability in the antioxidant and phenolic compounds between the species studied.</p>
			<p><bold>Isolation and identification of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> Out of the Platino variety of serrano peppers with symptoms of wilting, <italic>R. solani</italic> and <italic>P. aphanidermatum</italic> were isolated and identified. In <italic>R. solani</italic> plae maroon hyphae were observed, 5-8 µm in width and branched in a 90 ° angle (<xref ref-type="bibr" rid="B43">Watanabe and Matsuda, 1966</xref>), whereas for <italic>P. aphanidermatum</italic>, coenocytic mycelium, sporangia and oogonia were observed (Díaz <italic>et al</italic>., 2011), which correspond to the species <italic>R. solani</italic> and <italic>P. aphanidermatum</italic>, respectively. Regarding molecular identification, the sequences provided by the LANBAMA of the IPICYT for each species were compared with the BLAST of the National Center for Biotechnology Information. In the case of <italic>R. solani</italic>, its sequence obtained a coverage of 100% and 100% similarity when compared with accession number JX535004.1. On the other hand, the sequence for <italic>P. aphanidermatum</italic> obtained a coverage of 100% and 100% similarity when compared with accession number EU245039.1. Finally, both sequences were deposited in the GenBank, obtaining accession numbers PQ571162 for <italic>R. solani</italic>, and PQ571194 for <italic>P. aphanidermatum</italic>.</p>
			<p><bold>Antifungal activity of <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts on the mycelial growth of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> The results of the plant extracts on the mycelial growth of <italic>P. aphanidermatum</italic> displayed a highly significant effect. <italic>F. splendens</italic> was the most effective, with an inhibition of 100% from 250 mg L<sup>-1</sup>, whereas <italic>A. striata</italic> reached 100% inhibition at 1000 mg L<sup>-1</sup> (<xref ref-type="fig" rid="f1">Figure 1</xref>). The IC50 values were 61.2 mg L<sup>-1</sup> for <italic>Fouquieria splendens</italic> and 199.2 mg L<sup>-1</sup> for <italic>Agave striata</italic>, presenting statistical significance (<xref ref-type="table" rid="t3">Table 3</xref>). A study conducted by Milagrosa <italic>et al</italic>. (2007) reported that wine vinasse at concentrations of 5% inhibited <italic>P. aphanidermathum</italic> by 100%. Meanwhile, the results obtained from the plant extracts on the mycelial growth of <italic>R. solani</italic> displayed inhibition in the mycelial development of the pathogen. In turn, <italic>F. splendens</italic> presented an inhibition of 100% from 500 mg L<sup>-1</sup>, and <italic>A. striata</italic> reached an inhibition of 100% starting at 1000 mg L<sup>-1</sup> (<xref ref-type="fig" rid="f2">Figure 2</xref>). The lowest IC50 was 92.6 mg L<sup>-1</sup> for <italic>F. splendens</italic> and 211.2 mg L<sup>-1</sup> for <italic>A. striata,</italic> observing a statistical difference (<xref ref-type="table" rid="t3">Table 3</xref>). An investigation by Rodríguez <italic>et al</italic>. (2020) reported inhibitions of mycelial growth (IMG, %) in <italic>R. solani</italic> of 100 and 50% using <italic>Larrea tridentata</italic> and <italic>Rosmarinus oficinalis</italic> extracts, respectively.</p>
			<p>
				<fig id="f1">
					<label>Figure 1</label>
					<caption>
						<title>Inhibition of <italic>Pythium aphanidermatum</italic> with methanolic <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts using the poisoned medium method.</title>
					</caption>
					<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf1.gif"/>
				</fig>
			</p>
			<p>
				<table-wrap id="t3">
					<label>Table 3</label>
					<caption>
						<title>Inhibitory concentration at 50% (IC<sub>50</sub>) of methanolic <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts on <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani.</italic></title>
					</caption>
					<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
						<thead>
							<tr>
								<th/>
								<th style="font-style:italic">Pythium aphanidermatum</th>
								<th style="font-style:italic">Rhizoctonia solani</th>
							</tr>
						</thead>
						<tbody>
							<tr>
								<td style="font-style:italic">Agave striata</td>
								<td>199.17<sup>b</sup></td>
								<td>211.18<sup>b</sup></td>
							</tr>
							<tr>
								<td style="font-style:italic">Fouquieria splendens</td>
								<td>61.22<sup>a</sup></td>
								<td>92.59<sup>a</sup></td>
							</tr>
						</tbody>
					</table>
					<table-wrap-foot>
						<fn id="TFN2">
							<p>*Values with the same letter are statistically similar (Tukey, p&lt;0.05).</p>
						</fn>
					</table-wrap-foot>
				</table-wrap>
			</p>
			<p>
				<fig id="f2">
					<label>Figure 2</label>
					<caption>
						<title>Inhibition of <italic>Rhizoctonia solani</italic> using plant <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts using the poisoned medium method.</title>
					</caption>
					<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf2.gif"/>
				</fig>
			</p>
			<p>Caffeic acid, found in the <italic>A. striata</italic> and <italic>F. splendens</italic> extracts, has presented antifungal ability (<xref ref-type="bibr" rid="B11">Freires <italic>et al.</italic>, 2016</xref>), as observed in previous studies. An example of this is the study by <xref ref-type="bibr" rid="B39">Romero <italic>et al</italic>. (2009</xref>), who investigated the effect of phenolic compounds such as caffeic acid, rutin and quercetin in the growth of <italic>Aspergillus carbonarius</italic> and the production of ochratoxin A. They observed that all phenolic compounds, at a concentration of 250 mg L<sup>-1</sup>, had a significant impact on the growth rate of <italic>A. carbonarius</italic>. In addition, <xref ref-type="bibr" rid="B5">Bisogno <italic>et al</italic>. (2007</xref>) reported that the minimum inhibiting concentration (MIC) of caffeic acid for <italic>Aspergillus flavus</italic>, <italic>Aspergillus niger</italic> and <italic>Aspergillus terreus</italic> was greater than 250 mg L<sup>-1</sup>. On the other hand, the quercetin found in both extracts (<italic>A. striata</italic> and <italic>F. splendens</italic>) has displayed antifungal activity against five fungal species: <italic>Alternaria alternata, Aspergillus fumigatus</italic>, <italic>A. niger</italic>, <italic>Macrophomina phaseolina</italic> and <italic>Penicillium citrii</italic> (<xref ref-type="bibr" rid="B23">Kanwal <italic>et al</italic>., 2010</xref>). The flavonoid pinocembrin found in <italic>A. striata</italic> has anti-inflammatory, antimicrobial and antioxidant roles (<xref ref-type="bibr" rid="B36">Rasul <italic>et al</italic>., 2013</xref>) and has shown an inhibiting effect against <italic>Candida albicans</italic> (<xref ref-type="bibr" rid="B20">Hernández-Tasco <italic>et al</italic>., 2018</xref>; <xref ref-type="bibr" rid="B41">Tundis <italic>et al</italic>., 2018</xref>). The ellagitannin terflavin B, found in <italic>A. striata</italic>, has shown an inhibiting effect against <italic>C. albicans</italic> (Salih <italic>et al</italic>., 2022). In addition, the tannin pedunculagin, a phytochemical found in <italic>A. striata</italic>, exhibited an inhibiting effect against <italic>A. flavus</italic> in extractos <italic>Punica granatum</italic> extracts (Mostafa <italic>et al</italic>., 2011). <xref ref-type="bibr" rid="B21">Ilk <italic>et al</italic>. (2017</xref>) developed lecithin/chitosan nanoparticles loaded with kaempferol, in which they observed inhibitory activity against <italic>Fusarium oxysporum</italic>. However, pure kaempferol also proved to be effective against <italic>C. albicans</italic> (<xref ref-type="bibr" rid="B46">Yordanov <italic>et al</italic>., 2008</xref>). The flavonoids eriodyctiol, dihydroquercetin and luteolin, found in <italic>F. splendens</italic>, have displayed antifungal activity against <italic>Fusarium graminearum</italic> and <italic>Septoria zeicola</italic> in extracts of <italic>Ficus sarmentosa</italic> var<italic>. henryi</italic> after a fractionation process (<xref ref-type="bibr" rid="B42">Wang <italic>et al</italic>., 2010</xref>).</p>
			<p><bold>Effect of <italic>Agave striata</italic> and <italic>Fouquieria splendens</italic> extracts on the structures and morphology of <italic>Pythium aphanidermatum</italic> and <italic>Rhizoctonia solani</italic>.</bold> Both extracts displayed activity against <italic>P. aphanidermatum</italic> oogonia, leading to lysis at high doses of 2000 to 500 mgL<sup>-1</sup> and malformation and deformation of hyphae at concentrations of 199.2 and 61.2 mg L<sup>-1</sup> with the <italic>A. striata</italic> and <italic>F. splendens</italic> extracts (<xref ref-type="fig" rid="f3">Figure 3</xref>). <xref ref-type="bibr" rid="B27">Lópeza <italic>et al</italic>. (2022</xref>) proved that <italic>Bacillus amyleloquefaciens</italic> completely inhibited the germination of <italic>Phytophthora capsici</italic> zoospores, causing encystment, malformations in the germinational tube and cell degradation. The use of plant extracts altered the morphology of the <italic>R. solani</italic> hyphae, which appeared distorted and fragmented with concentrations of 211.2 mg L<sup>-1</sup> with the extract of</p>
			<p>
				<fig id="f3">
					<label>Figure 3</label>
					<caption>
						<title>Effect of plant extracts on <italic>Pythium aphanidermatum</italic> oogonia at 100X. A, B and C) <italic>Fouquieria splendens</italic>, D, E and F) <italic>Agave striata</italic>, and G) Negative control.</title>
					</caption>
					<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf3.gif"/>
				</fig>
			</p>
			<p><italic>A. striata</italic> and of 92.6 mg L<sup>-1</sup> with the <italic>F. splendens</italic> extract (<xref ref-type="fig" rid="f4">Figure 4</xref>), similar to descriptions by <xref ref-type="bibr" rid="B24">Khaledi <italic>et al</italic>. (2015</xref>) with <italic>Mentha piperita</italic>, <italic>Bunium persicum</italic> and <italic>Thymus vulgaris</italic> essential oils.</p>
			<p>The results can be attributed to the presence of antifungal compounds in the</p>
			<p><italic>A. striata</italic> and <italic>F. splendens</italic> extracts. For example, quercetin found in both extracts is a flavonoid that has been proven to reduce the content of ergosterol, a crucial sterol for the fluidity and stability of cell membranes, in <italic>Trichophyton rubrum</italic> (<xref ref-type="bibr" rid="B6">Bitencourt <italic>et al</italic>., 2013</xref>). It has also shown the inhibition of the enzyme CYP51, which is involved in the synthesis of steroids in fungi (<xref ref-type="bibr" rid="B25">Khanzada <italic>et al</italic>., 2021</xref>). The flavonoid pinocembrin, found in <italic>A. striata</italic>, has displayed different action mechanisms, including interference with energy homeostasis, increases in cell membrane permeability, reductions in the content of intracellular constituents (e.g., soluble proteins, reducing sugars and total lipids). Additionally, a marked reduction in chitin and glucanase contents was induced in the mycelia of <italic>Penicillium italicum</italic> (<xref ref-type="bibr" rid="B31">Peng <italic>et al</italic>., 2012</xref>; <xref ref-type="bibr" rid="B9">Chen <italic>et al</italic>., 2020</xref>). On the other hand, the flavonoid kaempferol found in <italic>F. splendens</italic> has been suggested in studies to cause damage to hyphae and is likely associated with cell wall alteration, leading to cell leakage and loss of cell integrity (<xref ref-type="bibr" rid="B21">Ilk <italic>et al</italic>., 2017</xref>). It has also shown synergistic antifungal activities with histone deacetylase (<xref ref-type="bibr" rid="B35">Rajasekharan <italic>et al</italic>., 2014</xref>), as well as the inhibition of the enzymes CYP51 and NDK, the latter being an enzyme involved in the phosphorylation of nucleosides and nucleotides (Khanzada <italic>et al</italic>., 2021).</p>
			<p>
				<fig id="f4">
					<label>Figure 4</label>
					<caption>
						<title>Effect of plant extracts on mycelial <italic>Rhizoctonia solani</italic> structures at 40X and 100X. A, B, C) <italic>Fouquieria splendens</italic>, D, E, F) <italic>Agave striata</italic>, and G), H) Negative control.</title>
					</caption>
					<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf4.gif"/>
				</fig>
			</p>
		</sec>
		<sec sec-type="conclusions">
			<title>Conclusions</title>
			<p>Phytochemicals with antifungal potential were identified in <italic>A. striata</italic> and <italic>F. splendens</italic>. Both extracts contained caffeic acid and quercetin. Additionally, <italic>A. striata</italic> displayed the presence of pinocembrin and terflavin B, whereas <italic>F. splendens</italic> presented eriodyctiol, kaempferol, dihydroquercetin, luteolin and pedunculagin. On the other hand, <italic>F. splendens</italic> was found to display a significantly higher antioxidant activity and CFT than <italic>A. striata</italic>. Both extracts displayed biological efficiency against <italic>P. aphanidermatum</italic> and <italic>R. solani</italic>. Likewise, they also presented alterations in the morphology of <italic>P. aphanidermatum</italic> oogina and the <italic>R. solani</italic> hyphae. These results indicate that <italic>A. striata</italic> and <italic>F. splendens</italic> extracts have promising antifungal compounds, suggesting its potential in future phytopathogen control studies.</p>
		</sec>
	</body>
	<back>
		<ack>
			<title>Acknowledgments</title>
			<p> </p>
			<p> </p>
			<p>To the Universidad Autónoma Agraria Antonio Narro and the Department of Agricultural Parasitology (project 2129), as well as to CONACYT (project 2019-000002-01NACF-00360) for their invaluable support and funds in this study.</p>
		</ack>
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	</back>
	<sub-article article-type="translation" id="s1" xml:lang="es">
		<front-stub>
			<article-categories>
				<subj-group subj-group-type="heading">
					<subject>Articles</subject>
				</subj-group>
			</article-categories>
			<title-group>
				<article-title>Efectividad biológica de extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> sobre <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani in vitro</italic></article-title>
			</title-group>
			<abstract>
				<title>Resumen:</title>
				<sec>
					<title>Antecedentes/Objetivo. </title>
					<p><italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic> son pa- tógenos que afectan cultivos agrícolas. Este estudio tuvo como objetivo evaluar extractos metanólicos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> del desierto chihu- ahuense sobre estos hongos, buscando alternativas de control biológico.</p>
				</sec>
				<sec>
					<title>Materiales y Métodos. </title>
					<p>Se aislaron los patógenos de plantas de chile, identificán- dolos por métodos morfológicos y moleculares. Se prepararon extractos metanó- licos de ambas plantas, y se analizó la capacidad antioxidante (CA), contenido de polifenoles totales (CPT), y compuestos antifúngicos por HPLC-MS. La efectivi- dad antifúngica se evaluó en concentraciones de 3.9 a 2000 mg L<sup>-1</sup> mediante medio envenenado y se observó el daño en las estructuras de los hongos.</p>
				</sec>
				<sec>
					<title>Resultados. </title>
					<p>Los extractos de <italic>F. splendens</italic> y <italic>A. striata</italic> mostraron una CA de 55% y 68%, respectivamente, y CPT de 61 mg/g y 112 mg/g. Ambos contenían áci- do cafeico y quercetina, mientras que <italic>F. splendens</italic> también presentaba eridictiol, kaempferol, y luteolina; <italic>A. striata</italic> contenía pinocembrina y terflavina B. <italic>F. splen- dens</italic> logró 100% de inhibición de <italic>P. aphanidermatum</italic> a 250 mg L<sup>-1</sup> y de <italic>R. solani</italic> a 500 mg L<sup>-1</sup>, mientras que <italic>A. striata</italic> alcanzó 100% de inhibición a 1000 mg L<sup>-1</sup> en ambos casos. Los extractos provocaron lisis en oogonios de <italic>P. aphanidermatum</italic> y fragmentación en el micelio de <italic>R. solani</italic>.</p>
				</sec>
				<sec>
					<title>Conclusión. </title>
					<p>Los extractos metanólicos de <italic>F. splendens</italic> y <italic>A. striata</italic> muestran acti- vidad antifúngica prometedora contra <italic>P. aphanidermatum</italic> y <italic>R. solani</italic>, sugiriendo que estos compuestos naturales pueden ser útiles como alternativa biológica para el control de estos patógenos en cultivos agrícolas.</p>
				</sec>
			</abstract>
			<kwd-group xml:lang="es">
				<title>Palabras clave:</title>
				<kwd>extractos vegetales</kwd>
				<kwd>efectividad biológica</kwd>
				<kwd>fitoquímicos</kwd>
				<kwd>hongos fitopatógenos<italic>.</italic></kwd>
			</kwd-group>
		</front-stub>
		<body>
			<sec sec-type="intro">
				<title>Introducción</title>
				<p><italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic> son patógenos ampliamente re- conocidos por su capacidad para causar enfermedades devastadoras en una varie- dad de cultivos agrícolas importantes. <italic>P. aphanidermatum</italic> causa pudrición de raíz, afecta el sistema radicular y provoca muerte de plántulas preemergentes por lesio- nes acuosas en raíz y tallo (<xref ref-type="bibr" rid="B33">Punja y Yip, 2003</xref>; <xref ref-type="bibr" rid="B15">Grijalba <italic>et al.</italic>, 2015</xref>). Por su parte <italic>R. solani</italic> afecta tallos desde la base, causando pudriciones blandas y daños radicu- lares que debilitan y matan las plantas prematuramente (<xref ref-type="bibr" rid="B29">Medeiros <italic>et al</italic>., 2015</xref>). La persistencia y adaptación de estos fitopatógenos en el suelo complican su manejo, afectando la seguridad alimentaria y la economía agrícola. Por lo anterior mencio- nado, en la actualidad se buscan alternativas sustentables, como los extractos vege- tales, destacando las plantas del Desierto Chihuahuense como una opción para la producción de extractos vegetales por su composición fitoquímica y su capacidad para inhibir el desarrollo de microorganismos fitopatógenos (Tucuch-Pérez <italic>et al</italic>., 2021). En este sentido <italic>Fouquieria splendens</italic> y <italic>Agave striata</italic> coexisten y se desarrollan dentro del Desierto Chihuahuense. <italic>A. striata</italic> o “agave espadín” tiene una amplia distribución en el sur, desde los estados mexicanos de Coahuila hasta Hidalgo (<xref ref-type="bibr" rid="B13">Gentry, 1982</xref>), en tanto que <italic>F. splendens</italic> u “ocotillo” se distribuye desde el suroes- te de Estados Unidos hasta la parte sur de Zacatecas, Querétaro e Hidalgo (Henrick- son, 1972). Investigaciones indican que plantas del Desierto Chihuahuense pueden inhibir o eliminar microorganismos fitopatógenos (Lira-Saldívar, 2003)<italic>,</italic> los cuales causan enfermedades devastadoras en cultivos de importancia económica (Martí- nez <italic>et al</italic>., 2016; Hernández <italic>et al</italic>., 2018).</p>
				<p>El control de microorganismos fitopatógenos se atribuye a los factores ambien- tales presentes en estas regiones, los cuales estimulan la producción de compuestos antimicrobianos en las plantas silvestres (Larios <italic>et al</italic>., 2020; Salas <italic>et al</italic>., 2023). Las familias Agavaceae y Fouquieriaceae han destacado por sus propiedades anti- microbianas (Gonzáles <italic>et al</italic>., 2015; Ramírez <italic>et al</italic>., 2023) . Actualmente, se conoce que <italic>A. striata</italic> y <italic>F. splendens</italic> se emplean para tratar diversas afecciones en la medi- cina tradicional (Pérez <italic>et al</italic>., 2003; López <italic>et al</italic>., 2022), y han mostrado propiedades antimicrobianas contra <italic>Clavibacter michiganensis</italic> subsp <italic>michiganensis</italic>, presentando fitoquímicos tales como alcaloides, carbohidratos, flavonoides, saponinas, taninos y quinonas (Ramírez <italic>et al</italic>., 2023). Los objetivos de la presente investigación fue- ron identificar compuestos fitoquímicos presentes en extractos metanólicos de <italic>A. striata</italic> y <italic>F. splendens</italic>, determinar su Capacidad Antioxidante (CA) y Contenido de Polifenoles Totales (CPT), evaluar su efectividad biológica <italic>in vitro</italic> sobre <italic>P. apha- nidermatum</italic> y <italic>R. solani,</italic> y observar cambios morfológicos en sus estructuras por los extractos vegetales.</p>
			</sec>
			<sec sec-type="materials|methods">
				<title>Materiales y métodos</title>
				<p>Colecta de plantas. Se colectaron hojas de <italic>A. striata</italic> y tallos de <italic>F. splendens</italic> en el mu- nicipio de General Cepeda, Coahuila, México (25°20’27.13’N y 101°27’52.07’’O a 1531 m.s.n.m.), durante el mes de agosto. Estas se depositaron en bolsas de plásti- co color negro para su traslado al laboratorio de Micología y Biotecnología del de- partamento de Parasitología Agrícola de la Universidad Autónoma Agraria Antonio Narro. El material vegetal se lavó con agua corriente, posteriormente se colocaron en una estufa de secado (Arsa AR-130D, México) a 60 °C hasta presentar peso constante. Finalmente, se trituraron con un molino (Surtek Mogra1, México); y se tamizaron con un tamiz de 0.2 mm para guardarse en recipientes oscuros debida- mente identificados hasta su uso.</p>
				<p>Preparación de los extractos. Los extractos se prepararon siguiendo la metodolo- gía reportada por Jasso <italic>et al</italic>. (2015), con algunas modificaciones, utilizando meta- nol al 96 % como solvente; se depositaron 42 g del polvo de cada planta en matra- ces de 500 mL adicionando 375 mL del solvente; y se colocaron en una parrilla de agitación (Thermo Scientific Cimarec, USA) durante 72 h a 60 °C. Posteriormente el solvente se separó mediante rota evaporación (IKA RV 10 digital V, USA) a 150 rpm a 70 °C. Una vez obtenida la fracción metanólica esta se vertió en recipientes de vidrio y se colocaron en una estufa de secado (Arsa AR-130D, México) a 50 °C durante 72 h, finalmente, los extractos se pulverización y se almacenaron en frascos color ámbar en refrigeración a temperatura de 4 °C.</p>
				<sec>
					<title><bold>Caracterización de fitoquímicos presentes en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> mediante Cromatografía de líquidos de fase inversa (HPLC-MS).</bold></title>
					<p>El análisis por cromatografía líquida de alta resolución en fase in- versa se realizó siguiendo la metodología de Ascacio <italic>et al</italic>. (2016), que consiste en utilizar un sistema de HPLC Varían que incluye un inyector automático (Varian ProStar 410, EE. UU.), una bomba ternaria (Varian ProStar 2310, EE. UU.) y un decantor PDA (Varian ProStar 330, EE. UU.). También se utilizó un espectrómetro de masas con trampa de iones de cromatógrafo de líquidos (Varian 500 - MS IT Mass Spectrometer, EE. UU.) equipado con una fuente de iones por electropul- verización. Se inyectaron muestras (5 μL) en una columna Denali C18 (150 mm x 2.1 mm, 3 μm, Grace, EE. UU.). La temperatura del horno se mantuvo a 30 °C. Los eluyentes fueron ácido fórmico (0.2 %, V/V; disolvente A) y acetontrilo (disol- vente B). Se aplicó el siguiente gradiente: inicial, 3 % B; 0 - 5 min, 9 % B lineal; 5 - 15 min, 16 % B lineal; 15 - 45 min, 50 % B lineal. Después la columna se lavó y se reacondicionó, el caudal se mantuvo a 0.2 mL/min y la elución se controló a 245, 280, 320 y 550 nm. Se inyectó todo el efluente (0.2 mL/min) en la fuente del espectrómetro de masas, sin dividir. Todos los experimentos de EM se llevaron a cabo en modo negativo [M-H]-1. Se utilizó nitrógeno como gas nebulizador y helio como gas amortiguador. Los parámetros de la fuente de iones fueron: voltaje de pulverización 5.0 kV y voltaje capilar y temperatura fueron 90.0 V y 350 °C, respectivamente. Los datos se recopilaron y procesaron utilizando el Software MS Workstation (V 6.9). Las muestras se analizaron en primer lugar en modo de barrido completo adquiridas en el rango m/z 50 - 2000. 6,951.6</p>
				</sec>
				<sec>
					<title><bold>Capacidad antioxidante DPPH en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic>.</bold></title>
					<p>Se determinó según lo descrito por Brand <italic>et al</italic>. (1995) con algunas modificaciones, se analizó por espectrofotometría, para ello se pesaron 50 mg de material seco al cual se le agregó 1.0 mL de metanol al 80 %. Posteriormente las muestras se zonificaron durante 20 min y centrifugaron 15 min a 14000 rpm. Una vez obtenido el extracto, se tomó una alícuota de 10 µL la cual fue colocada en microplacas de 96 pocillos y se añadió 290 µL del reactivo DPPH preparado con metanol absoluto al 100 µM. Posteriormente se dejó reaccionar la muestra en la os- curidad durante 30 min. Una vez transcurrido el tiempo la absorbancia fue medida a 515 nm. El porcentaje de inhibición DDPH de los extractos se determinó utilizando como testigo la absorbancia del metanol al 80 %. La CA DPPH se determinó me- diante la siguiente fórmula: Inhibición de DPPH (%) = Absorbancia del blanco /Absorbancia del extracto × 100</p>
					<p><bold>Capacidad de Polifenoles Totales en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic>.</bold> La CPT se determinó utilizando el método de Folin-Ciocalteu descrito por Singleton y Rossi (1965) con algunas modificaciones. En el análisis se tomó una alícuota de 10 µL del extracto de metanol al 80 %, posteriormente se agregó 790 µL de agua destilada, 50 µL de Folin y 150 µL de carbonato de sodio al 20 %. Posteriormente la muestra se dejó reposar 1 h en obscuridad, una vez trascurrido el tiempo la absorbancia fue medida en un lector de microplaca a 765 nm. El CPT se determinó usando una curva estándar con ácido gálico, y se expresó como GAE mg/g (miligramos equivalentes de ácido gálico por gramo).</p>
				</sec>
				<sec>
					<title><bold>Aislamiento e identificación de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>Se colectaron raíces y tallos de plantas de chile serrano (<italic>Capsicum annuum</italic>) Var. Platino en etapa de floración, con síntomas de marchitez, en el municipio de Escui- napa, Sinaloa, México, mediante un muestreo aleatorizado tomando 6 plantas por muestra, y se sembraron 5 muestras de tejido por caja Petri, realizando seis repeti- ciones. Las muestras se desinfectaron siguiendo la metodología de <xref ref-type="bibr" rid="B7">Booth (1977</xref>) se lavaron, seccionaron y sumergieron en cloro al 3 % durante 3 min, posteriormente se enjuagaron y secaron. Se sembraron en medio PDA y se incubaron a 28 °C por siete días. La purificación de los hongos se llevó a cabo mediante la técnica de punta de hifa, un método que permite obtener hongos axénicos o libres de contami- nantes. Posteriormente se realizó la identificación morfológica usando claves taxo- nómicas de Díaz <italic>et al</italic>. (2011) para la caracterización de <italic>P. aphanidermatum,</italic> y de <xref ref-type="bibr" rid="B43">Watanabe y Matsuda (1966</xref>) para <italic>R. solani</italic>. La identificación molecular la realizó el Laboratorio Nacional de Biotecnología Agrícola, Médica y Ambiental (LANBA- MA) del Instituto Potosino de Investigación Científica y Tecnológica (IPICYT). Utilizando los primers ITS1 e ITS4, los cuales amplifican la región ITS entre los genes de la subunidad ribosomal 18S y la subunidad ribosomal 28S, lo que permite una identificación específica de las especies fúngicas.</p>
				</sec>
				<sec>
					<title><bold>Actividad antifúngica de extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> so- bre el crecimiento micelial de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>La actividad antifúngica se determinó por el método de medio envenenado pro- puesto por Jasso <italic>et al</italic>. (2011); Se utilizaron los extractos de <italic>A. striata</italic> y <italic>F. splen- dens</italic> a concentraciones de 3.9 a 2000 mg L<sup>-1</sup>, además se utilizó un control negativo el cual consistió solo de medio de cultivo PDA sin extracto, se utilizaron cuatro repeticiones por concentración. Se colocaron discos de 0.4 mm de diámetro con micelio activo de <italic>P. aphanidermatum</italic> y <italic>R. solani</italic> de siete días de crecimiento en el medio envenenado; y se incubaron a 28 ± 2 °C. La evaluación se realizó midiendo el crecimiento radial de los hongos diariamente, y concluyó, cuando el control ne- tativo cubrió la placa Petri por completo, repitiéndose el experimento dos veces. El porcentaje de inhibición se determinó mediante la siguiente fórmula:</p>
					<p>Porcentaje de Inhibición= (DC-DT/DC) *100.</p>
					<p>Donde DC es el diámetro del tratamiento control, y DT es el diámetro de las dife- rentes concentraciones.</p>
				</sec>
				<sec>
					<title><bold>Efecto de extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> sobre estructuras y morfología de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>Se utilizó la metodología de <xref ref-type="bibr" rid="B24">Khaledi <italic>et al</italic>. (2015</xref>) con modificaciones para evaluar los cambios morfológicos inducidos por los extractos en oogonios de <italic>P. aphaniderma- tum</italic> e hifas de <italic>R. solani</italic>, usando los resultados de la concentración inhibitoria al 50% (CI<sub>50</sub>) de <italic>A. striata</italic> y <italic>F. splendens</italic>. Para obtener oogonios de <italic>P. aphanidermatum</italic>, se empleó el medio envenenado con las concentraciones de 199.17 y 61.22 mg L<sup>-1</sup> con los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic>, respectivamente, en medio de cultivo Agar V8 (20 g de agar, 3 g de CaCO3, 160 mL de jugo de verduras V8 Campbell’s y 840 mL de agua destilada), en tanto que para el crecimiento del micelio de <italic>R. solani</italic> se utilizó el medio de cultivo PDA en medio envenenado con los extractos a concentraciones de 211.18 y 92.59 mg L<sup>-1</sup>, en los dos experimentos se utilizó un control negativo sin extracto. Tras siete días de cultivo, se evaluaron los cambios morfológicos bajo microscopio óptico con el objetivo 40X y 100X, ambos experi- mentos se llevaron a cabo utilizando 6 repeticiones y se realizaron por duplicado.</p>
					<p>Análisis estadístico. Se realizó un análisis Probit de los ensayos <italic>in vitro</italic> para de- terminar la CI<sub>50</sub> de cada tratamiento utilizando el programa estadístico Statistical AnalysisSys, V9.0., con los resultados obtenidos se realizó análisis de varianza con las concentraciones inhibitorias; y se realizaron pruebas de Tukey (p&lt;0.05).</p>
				</sec>
			</sec>
			<sec sec-type="results|discussion">
				<title>Resultados y Discusión</title>
				<sec>
					<title><bold>Caracterización de fitoquímicos presentes en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> mediante cromatografía de líquidos de fase inversa (HPLC-MS).</bold></title>
					<p>Los fitoquímicos identificados en los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic> se pueden observar en el <xref ref-type="table" rid="t4">Cuadro 1</xref>. En el caso de <italic>A. striata</italic> se observó la presencia de dos elegitaninos (pedunculagina y terflavina B), un ácido hidroxi- cinámico (ácido cafeico), un metoxiflavonol (brasidina), un flavonol (quercetina), una flavonona (pinocepmbrina), y un terpeno fenólico (rosmadial). En <italic>A. striata</italic> se han reportado diversos compuestos bioactivos, incluyendo flavonoles (derivados de quercetina) (Almaraz <italic>et al</italic>., 2013), saponinas triterpenoides y esteroidales, así como alcaloides, carbohidratos, flavonoides (flavononas y chalconas), saponinas (triterpenoides y esteroidales), taninos (fenoles), quinonas (antraquinonas y benzo- quinonas) y cumarinas (Ramírez <italic>et al</italic>., 2023). En el caso de <italic>F. splendens</italic> se detectó la presencia de dos flavonoles (kaempferol y quercetina), dos flavones (roifolina y leutina), dos metoxiflavonoles (patuletina y dimetilquercetina), una flavanona (eriodictiol), un ácido hidroxicinámico (ácido cafeico), un dihidroflavonol (dihi- droquercetina), y otro polifenol (Phlorin). En este sentido Nevárez-Prado <italic>et al</italic>., 2021, mencionan que los fitoquímicos más característicos del género <italic>Fouquieria</italic> corresponden a flavonoides (leucocianidina, kaempferol y quercetina), los ácidos p-cumárico, cafeico, ferúlico, y elágico, así como cumarina y escopoletina. Por otra parte, Rodríguez (2010), encontró en <italic>F. splendens</italic> presencia de grupos carbonilo, oxidrilos fenólicos, esteroles y metilesteroles, cumarinas, saponinas, sesquiterpen- lactonas y flavonoides. En los tallos y hojas se han encontrado flavonoides como el kaempferol, ermanina y cetina, además flavonas apigenina, acacetina, luteolina y crisoeriol. Entre estos, apigenina y ermanina, rutina y quercetina-3-O-glucósido fueron los principales compuestos fenólicos en las hojas (<xref ref-type="bibr" rid="B45">Wollenweber, 1994</xref>; <xref ref-type="bibr" rid="B30">Monreal-García <italic>et al</italic>., 2019</xref>), también se han descubierto triterpenos, triterpenoi- des, saponinas esteroideas y compuestos iridoides (<xref ref-type="bibr" rid="B40">Takhtajan, 2009</xref>), y en las flores los fitoesteroles y alcanos de cadena larga y corta; y en el tallo compuestos como el Dammaran-triterpeno, el fouquierol y el isofouquierol, así como el dammarendiol en las raíces (<xref ref-type="bibr" rid="B19">Hegnauer, 1989</xref>; <xref ref-type="bibr" rid="B44">Waterman 1985</xref>). Por otro lado, se han aislado el pyxinol y el ocotillol, este último es una saponina triterpenoide el cual presenta actividades antimicrobianas (<xref ref-type="bibr" rid="B4">Bi <italic>et al</italic>., 2017</xref>). López <italic>et al</italic>. (2022) identificaron los principales fenoles en hojas de <italic>F. splendens</italic> como ácido gálico, ácido elágico y kaempferol-3-β-glucósido. Además, Ramírez <italic>et al</italic>. (2023) informaron que el extracto metanólico de tallos de <italic>F. splendens</italic> contiene alcaloides, carbohidratos, flavonoides (flavononas, flavonas, flavonoles y chalconas), azúcares reductores, saponinas (triterpenoides), taninos (derivados de ácido gálico, derivados de catecol y fenoles), y quinonas (antraquinonas y benzoquinonas). Las plantas dentro de su metabolismo producen compuestos con propiedades antimicrobianas, los cuales pueden controlar enfermedades causadas por microorganismos fitopatógenos. La extracción de estos compuestos y su análisis permiten su aplicación frente a diver- sos fitopatógenos, lo que ofrece una alternativa para el manejo de enfermedades en la agricultura (<xref ref-type="bibr" rid="B18">Hernández-Lauzardo <italic>et al</italic>., 2007</xref>). En este sentido, Ramírez <italic>et al</italic>. (2023) observaron que extractos de <italic>A. striata</italic> y <italic>F. splendens</italic> tuvieron actividad antimicrobiana contra <italic>Clavibacter michiganensis in vitro</italic> y bajo condiciones de invernadero. Siendo, el extracto de <italic>F. splendens</italic> el más efectivo al reducir la in- cidencia y severidad de la enfermedad. Los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic> han mostrado actividad antifúngica en más estudio (Sánchez <italic>et al</italic>., 2005; <xref ref-type="bibr" rid="B42">Wang <italic>et al</italic>., 2010</xref>), sin embargo, no se ha investigado específicamente su efecto antifúngico sobre <italic>R. solani</italic> y <italic>P. aphanidermatum</italic>.</p>
					<p>
						<table-wrap id="t4">
							<label>Cuadro 1</label>
							<caption>
								<title>Compuestos fitoquímicos identificados en extractos metanólicos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> caracteri- zados mediante cromatografía de líquidos de fase inversa (HPLC-MS).</title>
							</caption>
							<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
								<thead>
									<tr>
										<th>Especie</th>
										<th>Fitoquímico</th>
										<th>Tiempo de retención (min)</th>
										<th>Masa</th>
										<th>CAS</th>
										<th>Familia</th>
									</tr>
								</thead>
								<tbody>
									<tr>
										<td rowspan="7" style="font-style:italic">Agave striata</td>
										<td>Ácido cafeico 4-O-glucósido</td>
										<td>5.554</td>
										<td>341.1</td>
										<td>166735-99-5</td>
										<td>Ácidos hidroxicinámicos</td>
									</tr>
									<tr>
										<td>Pinocembrina</td>
										<td>21.192</td>
										<td>255.0</td>
										<td>480-39-7</td>
										<td>Flavanonas</td>
									</tr>
									<tr>
										<td>Pedunculagina II</td>
										<td>30.902</td>
										<td>785.1</td>
										<td>87687-52-3</td>
										<td>Elagitaninos</td>
									</tr>
									<tr>
										<td>Terflavina B</td>
										<td>35.531</td>
										<td>784.9</td>
										<td>103744-86-1</td>
										<td>Elagitaninos</td>
									</tr>
									<tr>
										<td>Brasidina</td>
										<td>40.341</td>
										<td>623.1</td>
										<td>17331-71-4</td>
										<td>Metoxiflavonoles</td>
									</tr>
									<tr>
										<td>Quercetina 3-O-glucurónido</td>
										<td>43.630</td>
										<td>477.2</td>
										<td>22688-79-5</td>
										<td>Flavonoles</td>
									</tr>
									<tr>
										<td>Rosmadial</td>
										<td>49.237</td>
										<td>343.9</td>
										<td>85514-31-4</td>
										<td>Terpenos fenólicos</td>
									</tr>
									<tr>
										<td rowspan="11" style="font-style:italic">Fouquieria splendens</td>
										<td>Ácido cafeico 4-O-glucósido</td>
										<td>5.525</td>
										<td>341.5</td>
										<td>166735-99-5</td>
										<td>Ácidos hidroxicinámicos</td>
									</tr>
									<tr>
										<td>Patuletina 3-O-glucosil-(1-&gt;6)-[apiosil(1-&gt;2)]-glucósido</td>
										<td>34.539</td>
										<td>787.1</td>
										<td>101021-30-1</td>
										<td>Metoxiflavonoles</td>
									</tr>
									<tr>
										<td>Kaempferol 3-O-xilosil-rutinósido</td>
										<td>35.796</td>
										<td>739.1</td>
										<td>31921-42-3</td>
										<td>Flavonoles</td>
									</tr>
									<tr>
										<td>Kaempferol 3-O-glucosil-ramnosil-glucósido</td>
										<td>37.228</td>
										<td>755.0</td>
										<td>136449-09-7</td>
										<td>Flavonoles</td>
									</tr>
									<tr>
										<td>Kaempferol 3-O-rutinósido</td>
										<td>37.924</td>
										<td>593.1</td>
										<td>17650-84-9</td>
										<td>Flavonoles</td>
									</tr>
									<tr>
										<td>3,7-dimetilquercetina</td>
										<td>39.085</td>
										<td>329.0</td>
										<td>2068-02-2</td>
										<td>Metoxiflavonoles</td>
									</tr>
									<tr>
										<td>Dihidroquercetina</td>
										<td>40.224</td>
										<td>303.0</td>
										<td>480-18-2</td>
										<td>Dihidroflavonoles</td>
									</tr>
									<tr>
										<td>Rhoifolina 4’-O-glucósido</td>
										<td>40.506</td>
										<td>739</td>
										<td>31498-83-6</td>
										<td>Flavones</td>
									</tr>
									<tr>
										<td>Luteolina 7-O-rutinósido</td>
										<td>41.399</td>
										<td>593</td>
										<td>20633-84-5</td>
										<td>Flavones</td>
									</tr>
									<tr>
										<td>Quercetina 3-O-glucurónido</td>
										<td>43.62</td>
										<td>480.2</td>
										<td>22688-79-5</td>
										<td>Flavonoles</td>
									</tr>
									<tr>
										<td>Eriodictiol</td>
										<td>43.803</td>
										<td>287.0</td>
										<td>552-58-9</td>
										<td>Flavanonas</td>
									</tr>
									<tr>
										<td>Florina</td>
										<td>47.363</td>
										<td>286.9</td>
										<td>28217-60-9</td>
										<td>Otros polifenoles</td>
									</tr>
								</tbody>
							</table>
						</table-wrap>
					</p>
				</sec>
				<sec>
					<title><bold>Capacidad antioxidante DPPH en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic>.</bold></title>
					<p>Los resultados mostraron que el extracto de <italic>F. splendens</italic> mostró una actividad antioxidante significativamente mayor en comparación con el extracto de <italic>A. striata</italic> (<xref ref-type="table" rid="t5">Cuadro 2</xref>). La inhibición del radical DPPH se expresó en porcentaje (%), siendo del 68.7 para <italic>F. splendens</italic> y del 55.9 para <italic>A. striata</italic>. En este sentido, <xref ref-type="bibr" rid="B12">Garza <italic>et al</italic>. (2012</xref>) determinaron que el extracto metanólico de <italic>F. splendens</italic> tenía una con- centración efectiva media (CE<sub>50</sub> considerada como la concentración de la muestra requerida para atrapar el 50 % de radicales DPPH libres) de 130.2 μg. López <italic>et al</italic>. (2022) determinaron la actividad antioxidante de <italic>F. splendens</italic> con resultados de DPPH (2430.1 µmol TE/g), Capacidad Antioxidante Equivalente a Trolox (TEAC) de 60.4 µmol TE/g, Capacidad de Absorción de Radicales Oxigenados (ORAC) de 4948.4 µmol TE/g y Poder de Reducción Férrica (FRAP) de 7803 µmol Fe(II)/g) (p &lt; 0.0001). En <italic>Agave</italic> ya hay investigaciones en la determinación de la capacidad antioxidante. En una investigación realizada por Ahumada <italic>et al</italic>. (2013) obtuvieron resultados que mostraron variaciones significativas en la actividad antioxidante en- tre diferentes especies de <italic>Agave</italic>. En <italic>Agave rzedowskiana</italic> presentó la mayor activi- dad antioxidante con 27.4 μM TE/g p.s., seguido de <italic>Agave ornithobroma</italic> con 19.9 μM TE/g p.s.</p>
					<p>
						<table-wrap id="t5">
							<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
								<thead>
									<tr>
										<th>Especie</th>
										<th>CA Porcentaje de Inhibición</th>
										<th>CPT CFT (GAE mg/g)</th>
									</tr>
								</thead>
								<tbody>
									<tr>
										<td style="font-style:italic">Agave striata</td>
										<td>55.90 ± 1.63<sup>a</sup></td>
										<td>61.2 ± 1.17<sup>a</sup></td>
									</tr>
									<tr>
										<td style="font-style:italic">Fouquieria splendens</td>
										<td>68.71 ± 1.22<sup>a</sup></td>
										<td>112.2 ± 3.69<sup>b</sup></td>
									</tr>
								</tbody>
							</table>
							<table-wrap-foot>
								<fn id="TFN3">
									<p>*Valores con la misma letra son estadísticamente similares (Tukey, p&lt;0.05).</p>
								</fn>
							</table-wrap-foot>
						</table-wrap>
					</p>
				</sec>
				<sec>
					<title><bold>Capacidad de Polifenoles Totales en extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic>.</bold></title>
					<p>Los resultados mostraron que el extracto de <italic>F. splendens</italic> exhibió po- lifenoles totales significativamente mayor en comparación con el extracto de <italic>A. striata</italic>. La CPT se expresó en valores de CFT (GAE mg/g), siendo 112.2 para <italic>F. splendens</italic> y 61.2 para <italic>A. striata</italic> (<xref ref-type="table" rid="t5">Cuadro 2</xref>). López <italic>et al</italic>. (2022) analizaron los compuestos fenólicos del extracto metanólico de hojas de <italic>F. splendens</italic> utilizando el método de Folin Ciocalteu y tras fraccionar, observaron que la fracción de ace- tato de etilo contenía la mayor cantidad de compuestos fenólicos, con 479.9 mg GAE/g (p &lt; 0,0001). En otro estudio, Ahumada <italic>et al</italic>. (2013) encontraron variacio- nes significativas en contenido total de fenoles entre diferentes especies de <italic>Agave</italic>. <italic>A. ornithobroma</italic> tuvo el valor más alto con 12.37 mg GAE/g p.s., mientras que <italic>Aagave angustifolia</italic> mostró el valor más bajo con 2.06 mg GAE/g p.s., indicando una variabilidad considerable en los compuestos antioxidantes y fenólicos entre las especies estudiadas.</p>
				</sec>
				<sec>
					<title><bold>Aislamiento e identificación de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>De las muestras de chile serrano variedad Platino con síntomas de marchitez, se ais- laron e identificaron <italic>R. solani</italic> y <italic>P. aphanidermatum</italic>. En <italic>R. solani</italic> se observaron hi- fas marrones pálido de 5-8 µm de ancho, ramificadas en ángulo de 90 °C (<xref ref-type="bibr" rid="B43">Watanabe y Matsuda, 1966</xref>), en tanto que para <italic>P. aphanidermatum</italic> se observó micelio ceno- cítico, esporangios y oogonios (Días <italic>et al</italic>., 2011), lo que corresponde a las especies <italic>R. solani</italic> y <italic>P. aphanidermatum</italic> respectivamente. En relación a la identificación molecular, las secuencias proporcionadas por el LANBAMA del IPICYT de cada especie, fueron comparadas en el BLAST del National Center for Biotechnology Information. En el caso de R<italic>. solani</italic>, su secuencia obtuvo un 100 % de cobertura y 100 % de similitud al compararse con el número de acceso JX535004.1. Por su parte, la secuencia de <italic>P. aphanidermatum</italic> obtuvo 100 % de cobertura y 100 % de similitud al compararse con el número de acceso EU245039.1. Finalmente, ambas secuencias fueron depositadas en el GenBank obteniendo los números de acceso PQ571162 para <italic>R. solani</italic>, y PQ571194 para <italic>P. aphanidermatum</italic>.</p>
				</sec>
				<sec>
					<title><bold>Actividad antifúngica de extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> sobre el crecimiento micelial de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>Los resultados de los extractos vegetales sobre el crecimiento micelial de</p>
					<p><italic>P. aphanidermatum</italic> mostraron un efecto altamente significativo. <italic>F. splendens</italic> fue el más efectivo con una inhibición del 100 % desde 250 mg L<sup>-1</sup>, mientras que <italic>A. striata</italic> alcanzó el 100 % de inhibición a 1000 mg L<sup>-1</sup> (<xref ref-type="fig" rid="f5">Figura 1</xref>). Los valores de CI fueron de 61.2 mg L<sup>-1</sup> para <italic>Fouquieria splendens</italic> y 199.2 mg L<sup>-1</sup> para <italic>Agave striata</italic>, mostrando significancia estadística (<xref ref-type="table" rid="t6">Cuadro 3</xref>). Un estudio realizado por Milagrosa <italic>et al</italic>. (2007) informaron que la vinaza de vino a concentraciones del 5 % inhibió al100 % a <italic>P. aphanidermathum</italic>. En tanto, los resultados obtenidos de los extractos vegetales sobre el crecimiento micelial de <italic>R. solani</italic> mostraron inhibición en el de- sarrollo micelial del patógeno. Por su parte <italic>F. splendens</italic> mostró una inhibición del 100 % desde 500 mg L<sup>-1</sup>, mientras que <italic>A. striata</italic> alcanzó el 100 % de inhibición desde 1000 mg L<sup>-1</sup> (<xref ref-type="fig" rid="f6">Figura 2</xref>). Las CI50 más baja fue de 92.6 mg L<sup>-1</sup> para <italic>F. splen- dens</italic> y 211.2 mg L<sup>-1</sup> para <italic>A. striata,</italic> observándose diferencia estadística (<xref ref-type="table" rid="t6">Cuadro 3</xref>). En una investigación realizada por Rodríguez <italic>et al</italic>. (2020) reportaron inhibiciones del crecimiento micelial (ICM, %) de <italic>R. solani</italic> del 100 y 50 % utilizando extractos de <italic>Larrea tridentata</italic> y <italic>Rosmarinus oficinalis</italic>, respectivamente. El ácido cafeico, presente en los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic>, ha mos- trado capacidad antifúngica (<xref ref-type="bibr" rid="B11">Freires <italic>et al.</italic>, 2016</xref>), como se ha observado en estudios previos. Un ejemplo de esto es el estudio realizado por <xref ref-type="bibr" rid="B39">Romero <italic>et al</italic>. (2009</xref>) inves- tigaron el efecto de compuestos fenólicos como el ácido cafeico, rutina y querceti- na en el crecimiento de <italic>Aspergillus carbonarius</italic> y la producción de ocratoxina A.</p>
					<p>
						<fig id="f5">
							<label>Figura 1. </label>
							<caption>
								<title>Inhibiciónde <italic>Pythium aphanidermatum</italic> por extractos metanólicos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> por el método de medio envenenado. </title>
							</caption>
							<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf5.gif"/>
						</fig>
					</p>
					<p>
						<fig id="f6">
							<label>Figura 2</label>
							<caption>
								<title>Inhibición de <italic>Rhizoctonia solani</italic> por extractos vegetales <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> por el método de medio envenenado.</title>
							</caption>
							<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf6.gif"/>
						</fig>
					</p>
					<p>
						<table-wrap id="t6">
							<label>Cuadro 3</label>
							<caption>
								<title>Concentración inhibitoria al 50 % (CI<sub>50</sub>) de extractos metanólicos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> sobre <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani.</italic></title>
							</caption>
							<table border="1" cellpadding="5" cellspacing="0" style="border-collapse: collapse; width: 100%; text-align: center;">
								<thead>
									<tr>
										<th> </th>
										<th style="font-style:italic">Pythium aphanidermatum</th>
										<th style="font-style:italic">Rhizoctonia solani</th>
									</tr>
								</thead>
								<tbody>
									<tr>
										<td style="font-style:italic">Agave striata</td>
										<td>199.17<sup>b</sup></td>
										<td>211.18<sup>b</sup></td>
									</tr>
									<tr>
										<td style="font-style:italic">Fouquieria splendens</td>
										<td>61.22<sup>a</sup></td>
										<td>92.59<sup>a</sup></td>
									</tr>
								</tbody>
							</table>
							<table-wrap-foot>
								<fn id="TFN4">
									<p>*Valores con la misma letra son estadísticamente similares (Tukey, p&lt;0.05).</p>
								</fn>
							</table-wrap-foot>
						</table-wrap>
					</p>
					<p>Observaron que todos los compuestos fenólicos, a una concentración de 250 mg L<sup>-1</sup>, tuvieron un impacto significativo en la tasa de crecimiento de <italic>A. carbonarius</italic>. Además, <xref ref-type="bibr" rid="B5">Bisogno <italic>et al</italic>. (2007</xref>) reportaron que la concentración mínima inhibidora (CIM) del ácido cafeico para <italic>Aspergillus flavus</italic>, <italic>Aspergillus niger</italic> y <italic>Aspergillus terreus</italic> fue superior a 250 mg L<sup>-1</sup>. Por otro lado, la quercetina presente en ambos extractos (<italic>A. striata</italic> y <italic>F. splendens</italic>) ha mostrado actividad antifúngica contra cinco especies de hongos: <italic>Alternaria alternata, Aspergillus fumigatus</italic>, <italic>A. niger</italic>, <italic>Macro- phomina phaseolina</italic> y <italic>Penicillium citrii</italic> (<xref ref-type="bibr" rid="B23">Kanwal <italic>et al</italic>., 2010</xref>). El flavonoide pino- cembrina encontrado en <italic>A. striata</italic> tiene un papel antiinflamatorio, antimicrobiano y antioxidante (<xref ref-type="bibr" rid="B36">Rasul <italic>et al</italic>., 2013</xref>), ha mostrado efecto inhibitorio contra <italic>Candida albicans</italic> (<xref ref-type="bibr" rid="B20">Hernández-Tasco <italic>et al</italic>. ,2018</xref>; <xref ref-type="bibr" rid="B41">Tundis <italic>et al</italic>., 2018</xref>). El elagitanino ter- flavina B, presente en <italic>A. striata</italic>, ha demostrado un efecto inhibitorio contra <italic>C. albicans</italic> (Salih <italic>et al</italic>., 2022). Además, se descubrió que el tanino pedunculagina, un fitoquímico presente en <italic>A. striata</italic>, exhibió efecto inhibitorio contra <italic>A. flavus</italic> en extractos de <italic>Punica granatum.</italic> (Mostafa <italic>et al</italic>., 2011). <xref ref-type="bibr" rid="B21">Ilk <italic>et al</italic>. (2017</xref>) desarrollaron nanopartículas de lecitina/quitosano cargadas con kaempferol en el cual observaron la actividad de inhibición contra <italic>Fusarium oxysporum</italic>. Sin embargo, kaempferol puro también demostró ser eficaz contra <italic>C. albicans</italic> (<xref ref-type="bibr" rid="B46">Yordanov <italic>et al</italic>., 2008</xref>). Los flavonoides eriodictiol, dihidroquercetina y luteolina, presentes en <italic>F. splendens</italic>, han demostrado actividad antifúngica contra <italic>Fusarium graminearum</italic> y <italic>Septoria zeicola</italic> en el extracto de <italic>Ficus sarmentosa</italic> var<italic>. henryi</italic> tras un proceso de fracciona- miento (<xref ref-type="bibr" rid="B42">Wang <italic>et al</italic>., 2010</xref>).</p>
				</sec>
				<sec>
					<title><bold>Efecto de extractos de <italic>Agave striata</italic> y <italic>Fouquieria splendens</italic> sobre estructuras y morfología de <italic>Pythium aphanidermatum</italic> y <italic>Rhizoctonia solani</italic>.</bold></title>
					<p>Ambos extractos mostraron actividad contra los oogonios de <italic>P. aphanidermatum</italic>, provocando lisis a dosis altas de 2000 a 500 mgL<sup>-1</sup>, y malformación y deformación de hifas a con- centraciones de 199.2 y 61.2 mg L<sup>-1</sup> con los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic> (<xref ref-type="fig" rid="f7">Figura 3</xref>). <xref ref-type="bibr" rid="B27">Lópeza <italic>et al</italic>. (2022</xref>) probaron que lipopéptidos de <italic>Bacillus amyleloque- faciens</italic> inhibieron completamente la germinación de zoosporas de <italic>Phytophthora capsici</italic>, causando enquistamiento, malformaciones en el tubo germinal y degrada- ción celular. El uso de extractos vegetales alteró la morfología de las hifas de <italic>R. solani</italic>, que aparecían distorsionadas y fragmentadas con concentraciones de 211.2 mg L<sup>-1</sup> con el extracto de <italic>A. striata</italic> y de 92.6 mg L<sup>-1</sup> con el extracto de <italic>F. splendens</italic> (<xref ref-type="fig" rid="f8">Figura 4</xref>), similar a lo descrito por <xref ref-type="bibr" rid="B24">Khaledi <italic>et al</italic>. (2015</xref>) con aceites esenciales de <italic>Mentha piperita</italic>, <italic>Bunium persicum</italic> y <italic>Thymus vulgaris</italic>.</p>
					<p>Los resultados pueden atribuirse a la presencia de compuestos antifúngicos en los extractos de <italic>A. striata</italic> y <italic>F. splendens</italic>. Por ejemplo, la quercetina presente en ambos extractos es un flavonoide, que ha demostrado reducir el contenido de er- gosterol, un esterol crucial para la fluidez y estabilidad de las membranas celula- res, en <italic>Trichophyton rubrum</italic> (<xref ref-type="bibr" rid="B6">Bitencourt <italic>et al</italic>., 2013</xref>), así también ha mostrado inhibición de la enzima CYP51, que está involucrada en la síntesis de esteroides en hongos (<xref ref-type="bibr" rid="B25">Khanzada <italic>et al</italic>., 2021</xref>). El flavonoide pinocembrina encontrado en <italic>A. striata</italic> ha mostrado diferentes mecanismos de acción, entre los que se encuentran; la interferencia en la homeostasis energética, aumentos en la permeabilidad de la membrana celular, disminuciones en el contenido de los constituyentes intracelu- lares (proteínas solubles, azúcares reductores y lípidos totales), además, se indujo una marcada disminución en los contenidos de quitina y glucanasa de los micelios de <italic>Penicillium italicum</italic> (<xref ref-type="bibr" rid="B31">Peng <italic>et al</italic>., 2012</xref>; <xref ref-type="bibr" rid="B9">Chen <italic>et al</italic>., 2020</xref>). Por otro lado, el flavonoide kaempferol encontrado en <italic>F. splendens</italic> ha sido sugerido en estudios por causar daños en las hifas y probablemente esté asociado con la alteración de la pared celular, lo que provoca fugas celulares y pérdida de integridad celular (<xref ref-type="bibr" rid="B21">Ilk <italic>et al</italic>., 2017</xref>), también ha mostrado actividades antifúngicas sinérgicas de la histo- na desacetilasa (<xref ref-type="bibr" rid="B35">Rajasekharan <italic>et al</italic>., 2014</xref>), así también inhibición de las enzimas CYP51 y NDK la cual es una enzima implicada en la fosforilación de nucleósidos y nucleótidos (Khanzada <italic>et al</italic>., 2021).</p>
					<p>
						<fig id="f7">
							<label>Figura 3</label>
							<caption>
								<title>Efecto de extractos vegetales sobre oogonios de <italic>Pythium aphanidermatum</italic> a 100X. A, B y C) <italic>Fouquieria splendens</italic>, D, E y F) <italic>Agave striata</italic>, y G) Control negativo.</title>
							</caption>
							<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf7.gif"/>
						</fig>
					</p>
				</sec>
			</sec>
			<sec sec-type="conclusions">
				<title>Conclusiones</title>
				<p>Se identificaron fitoquímicos con potencial antifúngico en <italic>A. striata</italic> y <italic>F. splen- dens</italic>. Ambos extractos contenían ácido cafeico y quercetina. Además, <italic>A. striata</italic> mostró presencia de pinocembrina y terflavina B, mientras que <italic>F. splendens</italic> pre- sentó eridictiol, kaempferol, dihidroquercetina, luteolina y pedunculagina. Por otro lado, se encontró que <italic>F. splendens</italic> exhibía una actividad antioxidante y CFT significativamente mayor que <italic>A. striata</italic>. Ambos extractos demostraron efectividad biológica contra <italic>P. aphanidermatum</italic> y <italic>R. solani</italic>. Así también estos mostraron alte- raciones en la morfología de los oogonios de <italic>P. aphanidermatum</italic> y de las hifas de <italic>R. solani</italic>. Estos resultados indican que extractos de <italic>A. striata</italic> y <italic>F. splendens</italic> poseen compuestos antifúngicos prometedores, sugiriendo su potencial en futuros estudios de control de fitopatógenos.</p>
				<p>
					<fig id="f8">
						<label>Figura 4</label>
						<caption>
							<title>Efecto de extractos vegetales sobre estructuras miceliales de <italic>Rhizoctonia solani</italic> a 40X y 100X. A, B, C) <italic>Fouquieria splendens</italic>, D, E, F) <italic>Agave striata</italic>, y G), H) Control negativo.</title>
						</caption>
						<graphic xlink:href="2007-8080-rmfi-42-04-RMEXFIT202423-gf8.gif"/>
					</fig>
				</p>
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				<title>Agradecimientos</title>
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				<p>A la Universidad Autónoma Agraria Antonio Narro y al Departamento de Parasitología Agrícola (proyecto 2129), así como al CONACYT (proyecto 2019-000002-01NACF-00360) por su invaluable apoyo y financiamiento en este estudio.</p>
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