Rhizospheric Fungal spp. of Tomato (Solanum lycopersicum L.) and Their Effect on Tomato Plant Growth and on Alternaria solani, the Causative Agent of Early Blight of Tomato

Deshappriya N., Fernando W.C.J.O., Fernando M.S.W.
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Abstract

Tomato (Solanum lycopersicum L.) is a widely consumed crop throughout the world including Sri Lanka. Use of agrochemicals for enhanced yields and managing diseases associated with tomato results in deleterious impacts on soil microflora, fauna and leads to human health problems. A possible alternative is the use of rhizospheric microorganisms, as they are reported to improve productivity and protect various crops. Hence this study aimed to isolate rhizospheric fungal assemblages of tomato plants and evaluate their ability to solubilize phosphate while, controlling Alternaria solani, the causative agent of Early Blight of tomatoes. Rhizospheric fungi were isolated from commercial tomato cultivations in two agro-ecological zones (WM3b and IM2b), using soil dilution plate technique. Composite samples of soil attached to the root systems of five healthy plants, uprooted from each field were prepared and 10 g of each sample was used for the isolations. Twenty-seven fungal genera were isolated and Trichoderma and Penicillium were the frequently isolated genera. Phosphate solubilization ability of each fungal species was evaluated using Pikovskaya's agar and liquid medium, and four isolates (Eupenicillium sp., Talaromyces purpureogenus, Paecilomyces sp., and Purpureocillium lilacinum) with the highest phosphate solubilizing abilities were evaluated on tomato plant growth under greenhouse condition. The inocula of each fungal sp. were prepared by adding fifteen, 6mm diameter mycelial discs from each 5-day-old culture into polypropylene bags containing 90 g of sterile rice husk medium and incubated for 02 weeks at room temperature. Each pot contained a 4.6 kg layer of non- sieved sterilized potting medium (Sand: topsoil: Compost at 2:1:1 ratio) and a 2.5 kg layer of sieved medium on top, which was mixed with 15 g of inoculum. Three-week old, healthy tomato seedlings (variety-Bathiya) were planted (with 3 replicates) and allowed to grow with regular watering. Control pots contained potting medium with uninoculated sterile rice husk. Growth parameters such as Root and shoot lengths, fresh and dry weights of roots, shoots, and leaves were evaluated at 02-week intervals for 02 months and the results were analysed using ANOVA with Tukey‘s pairwise comparison. Plants treated with T. purpureogenus and P. lilacinum showed significant increases (p≤0.05) in all growth parameters in every evaluation. Dual culture plate assay on the Potato Dextrose Agar was used to test the effect of rhizospheric fungal spp. on the radial colony growth of A. solani, and Eupenicillium sp. showed the highest percentage inhibition (85.09±5.68%). Microscopic observations of inhibition zones between two colonies showed the presence of coils, loops, knobs, and haustoria, formed by inhibitory rhizospheric fungal species. Assays for Chitinase and Glucanase enzyme production showed that Mortierella sp.-1, P. lilacinum, Trichoderma sp.-1, and Acremonium sp. were the highest chitinase producers, whereas P. lilacinum was the highest glucanase producer. These results indicate the ability of some rhizospheric fungal species of tomato to increase the growth of tomato plants significantly by phosphate solubilization and ability to inhibit the growth of foliar pathogen A. solani under in vitro conditions.  Keywords: Rhizospheric fungi, Phosphate solubilization, Dual culture plate, Plant growth parameters 
番茄(Solanum lycopersicum L.)根瘤菌属及其对番茄植株生长和番茄早疫病病原菌Alternaria solani的影响
番茄(Solanum lycopersicum L.)是包括斯里兰卡在内的全世界广泛食用的作物。为提高产量和防治番茄病害而使用农用化学品会对土壤微生物区系和动物区系造成有害影响,并导致人类健康问题。一种可能的替代方法是使用根瘤微生物,因为据报道它们可以提高产量并保护各种作物。因此,本研究旨在分离番茄植株根瘤菌群,并评估其溶解磷酸盐的能力,同时控制番茄早疫病的病原体 Alternaria solani。利用土壤稀释板技术,从两个农业生态区(WM3b 和 IM2b)的商业番茄种植区分离出根瘤菌。从每块田地中拔出五株健康植株的根系,制备附着在其根系上的土壤复合样本,每个样本 10 克用于分离。共分离出 27 个真菌属,其中毛霉和青霉是最常分离出的真菌属。使用 Pikovskaya 琼脂和液体培养基对各真菌菌种的磷酸盐溶解能力进行了评估,并在温室条件下对番茄植株的生长情况进行了评估,结果表明 4 个分离菌种(大戟霉属、紫红塔拉酵母菌属、白疫霉属和紫青霉属)的磷酸盐溶解能力最高。每种真菌的接种体都是在装有 90 克无菌稻壳培养基的聚丙烯袋中加入 15 个直径为 6 毫米的菌丝盘,然后在室温下培养 02 周。每个花盆中都有一层 4.6 千克的未过筛灭菌盆栽培养基(沙:表土:堆肥,比例为 2:1:1),上面还有一层 2.5 千克的过筛培养基,培养基中混有 15 克接种物。种植三周大的健康番茄幼苗(品种-Bathiya)(3 个重复),并定期浇水让其生长。对照盆中的盆栽介质是未接种的无菌稻壳。在 02 个月的时间里,每隔 02 周对根长和芽长、根、芽和叶的鲜重和干重等生长参数进行评估,并使用方差分析和 Tukey 配对比较法对结果进行分析。在每次评估中,用紫花酢浆草和紫花酢浆草处理过的植物的所有生长参数都有显著增加(p≤0.05)。在马铃薯葡萄糖琼脂上使用双培养板试验来检测根瘤真菌属对茄属真菌径向菌落生长的影响,结果表明 Eupenicillium sp.的抑制率最高(85.09±5.68%)。通过显微镜观察两个菌落之间的抑制区,可以发现抑制性根瘤真菌菌种形成的线圈、环、节和菌丝体。几丁质酶和葡聚糖酶的产量测定显示,莫蒂尔氏菌(Mortierella sp.-1)、丁香菌(P. lilacinum)、毛霉菌(Trichoderma sp.-1)和Acremonium sp.的几丁质酶产量最高,而丁香菌(P. lilacinum)的葡聚糖酶产量最高。这些结果表明,在离体条件下,番茄根瘤菌中的一些菌种能通过磷酸盐溶解作用显著提高番茄植株的生长速度,并能抑制叶面病原菌A. solani的生长。 关键词:根瘤真菌根瘤真菌 磷酸盐溶解 双培养板 植物生长参数
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