Yang Fan, Abdelnabby Hazem, Xiao Yannong
Key Laboratory of Plant Pathology of Hubei Province, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Henan Academy of Agricultural Sciences, Zhengzhou, Henan Province, 450002, China.
Key Laboratory of Plant Pathology of Hubei Province, Huazhong Agricultural University, Wuhan, Hubei 430070, China; Department of Plant Protection, Faculty of Agriculture, Benha University, Qaliubia 13736, Egypt.
Microb Pathog. 2015 Dec;89:169-76. doi: 10.1016/j.micpath.2015.10.012. Epub 2015 Oct 28.
Sclerotinia sclerotiorum causes severe stem rot and yield loss in oilseed rape (Brassica napus L.) and other crops worldwide. Extensive studies have been conducted on Paecilomyces lilacinus as a nematophagous bioagent. However, no reports stated the effect of P. lilacinus as a biocontrol agent against oilseed rape rot S. sclerotiorum. This study describes such effect in lab and field trials using the new transformant pt361 derived from the wild strain P. lilacinus 36-1. Unlike the wild-type strain, the mutant pt361 showed high antagonistic effect against S. Sclerotiorum A. Under lab conditions, the pt361 inhibited (65%) radial mycelial growth of S. sclerotiorum in dual culture test producing 5.9 mm inhibition zone IZ in front of the S. sclerotiorum colony. Moreover, the cell-free filtrate of pt361 culture showed strong inhibitory effects (60.3-100%) on mycelial growth of S. sclerotiorum. In leaf detached assay, pt361 significantly (p < 0.05) inhibited (40.4-97.9%) the extension of the leaf spots caused by S. sclerotiorum A at all tested concentrations. The genomic DNA sequences of the inserted T-DNA flanking obtained from pt361 strain was cloned, verified as a glycoside hydrolase 31 family by homologous analysis with other fungal strains, and named PGH31 (2556bp). Secondary structure prediction showed a domain (Glycoside hydrolase31). Three years field trial confirmed that the cell-free filtrates or spores suspension of pt361 achieved significant (p < 0.05) suppression of oilseed rape stem rot, promoted growth and increased yield compared to the control and exceeded, at dose 100%, the action of the fungicide procymidone(®). In conclusion, the mutant pt361 of P. lilacinus is a novel and promising biocontrol agent against oilseed rape Sclerotinia stem rot.
核盘菌在全球范围内导致油菜(甘蓝型油菜)和其他作物严重的茎腐病并造成产量损失。人们对淡紫拟青霉作为一种食线虫生物制剂进行了广泛研究。然而,尚无报道说明淡紫拟青霉作为生物防治剂对油菜菌核病的防治效果。本研究通过使用源自野生菌株淡紫拟青霉36-1的新转化体pt361,在实验室和田间试验中描述了这种效果。与野生型菌株不同,突变体pt361对核盘菌A表现出高拮抗作用。在实验室条件下,pt361在双培养试验中抑制了核盘菌65%的径向菌丝生长,在核盘菌菌落前方产生了5.9毫米的抑菌圈IZ。此外,pt361培养物的无细胞滤液对核盘菌的菌丝生长表现出强烈的抑制作用(60.3%-100%)。在离体叶片试验中,pt361在所有测试浓度下均显著(p<0.05)抑制了核盘菌A引起的叶斑扩展(40.4%-97.9%)。从pt361菌株获得的插入T-DNA侧翼的基因组DNA序列被克隆,通过与其他真菌菌株的同源分析验证为糖苷水解酶31家族,并命名为PGH31(2556bp)。二级结构预测显示有一个结构域(糖苷水解酶31)。三年的田间试验证实,与对照相比,pt361的无细胞滤液或孢子悬浮液对油菜茎腐病有显著(p<0.05)的抑制作用,促进了生长并提高了产量,在剂量为100%时超过了杀菌剂腐霉利(®)的防治效果。总之,淡紫拟青霉突变体pt361是一种新型且有前景的防治油菜菌核病茎腐病的生物防治剂。