Antifungal activity of Leonotis nepetifolia extracts against Colletotrichum species causing bean anthracnose and their phytochemical analysis using LC-MS

Faustine Christopher , Patrick Ndakidemi , Samuel Nyalala , Ernest Mbega
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Abstract

Bean anthracnose results in significant yield losses, and the present primary management strategy of using synthetic fungicides is hampered by a number of factors. This study assessed the antifungal effects of aqueous, methanolic, and ethyl acetate extracts of Leonotis nepetifolia (Lamiaceae) against Colletotrichum species that cause bean anthracnose. The poisoned food technique was used to evaluate extracts in-vitro, and trials were carried out on farmer's plots. On the other hand, phytochemical analysis was done using LC-MS. Results obtained in-vitro show that the ethyl acetate extract was the most effective since it completely inhibited mycelial development at concentrations of 5 mg/mL and 2.5 mg/mL. On the other hand, the aqueous and methanolic extracts exhibited a comparable inhibitory effect, and at the higher concentration (5.0 mg/mL), a remarkable percentage of inhibition of 73.3% and 83.1% was recorded. According to field trials, bean anthracnose incidence was low and there were fewer infected pods per plant in plots treated with L. nepetifolia extracts than in the negative control. In comparison to the negative control plots, extracts-treated plots had larger weights of 100 seeds but no discernible negative effect in seed germination. LC-MS analysis indicated the presence of Linoleoyl ethanolamide, Curcumol, Deguelin, 9,10-dihydroxy-12Z-octadecenoic acid, Naringenin-7-O-glucoside, and 9S-hydroxy-10e 12z 15z-octadecatrienoic acid in the ethyl acetate extract whereas 6-Methoxyluteolin, Genistein, 5,7,3,4-tetramethoxyisoflavone, and 3,4-Dihydroxymandelic acid were identified in the aqueous extract. Antifungal activity and the presence of bioactive components revealed by extracts in this study demonstrate the potential of L. nepetifolia as a source of botanical fungicide for the management of bean anthracnose.

Leonotis nepetifolia提取物对引起豆炭疽病的炭疽菌的抑菌活性及其LC-MS分析
豆炭疽病导致显著的产量损失,目前使用合成杀菌剂的主要管理策略受到许多因素的阻碍。本研究评估了裙带菜(Lamiaceae)的水提取物、甲醇提取物和乙酸乙酯提取物对引起豆炭疽病的炭疽菌的抗真菌作用。采用中毒食品技术对提取物进行了体外评价,并在农户小区进行了试验。另一方面,使用LC-MS进行植物化学分析。体外结果表明,乙酸乙酯提取物在5 mg/mL和2.5 mg/mL浓度下完全抑制菌丝生长,因此效果最好。另一方面,水提取物和甲醇提取物表现出相当的抑制效果,在更高浓度(5.0 mg/mL)下,抑制率分别为73.3%和83.1%。根据田间试验,与阴性对照相比,用荆芥提取物处理的地块中,大豆炭疽病的发病率较低,单株受感染的荚数较少。与阴性对照地块相比,提取物处理地块的100粒种子重量更大,但对种子发芽没有明显的负面影响。LC-MS分析表明,乙酸乙酯提取物中存在亚麻酰乙醇酰胺、姜黄醇、Deguelin、9,10-二羟基-12Z-十八碳烯酸、Naringen-7-O-葡萄糖苷和9S-羟基-10e 12Z-15z-十八碳三烯酸,而水提取物中鉴定出6-甲氧基uteolin、Genistein、5,7,3,4-四甲氧基异黄酮和3,4-二羟基扁桃酸。本研究中提取物显示的抗真菌活性和生物活性成分的存在证明了荆芥作为植物杀菌剂来源治疗豆炭疽病的潜力。
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