The use of biological control agents (BCAs) is a promising alternative control measure for Fusarium crown rot (FCR) of wheat caused by Fusarium pseudograminearum. A bacterial strain, YB-185, was isolated from the soil of wheat plants with FCR and identified as Bacillus velezensis. YB-185 exhibited strong inhibition of F. pseudograminearum mycelial growth and conidial germination in culture. Seed treatment with YB-185 in greenhouse and field resulted in reductions in disease by 66.1% and 57.6%, respectively, along with increased grain yield. Microscopy of infected root tissues confirmed that YB-185 reduced root invasion by F. pseudograminearum. RNA-seq of F. pseudograminearum during co-cultivation with B. velezensis YB-185 revealed 5086 differentially expressed genes (DEGs) compared to the control. Down-regulated DEGs included genes for glucan synthesis, fatty acid synthesis, mechanosensitive ion channels, superoxide dismutase, peroxiredoxin, thioredoxin, and plant-cell-wall-degrading enzymes, whereas up-regulated DEGs included genes for chitin synthesis, ergosterol synthesis, glutathione S-transferase, catalase, and ABC transporters. In addition, fungal cell apoptosis increased significantly, as indicated by TUNEL staining, and the scavenging rate of 2,2′-Azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt radical cation (ABTS·+) in the fungus significantly decreased. Thus, F. pseudograminearum may be trying to maintain normal cell functions by increasing cell wall and membrane synthesis, antioxidant and anti-stress responses, detoxification of bacterial antimicrobial compounds, and transportation of damaging compounds from its cells. However, cell death and free radical accumulation still occurred, indicating that the responses were insufficient to prevent cell damage. Bacillus velezensis YB-185 is a promising BCA against FCR that acts by directly damaging F. pseudograminearum, thus reducing its ability to colonize roots and produce symptoms.
Cereal cyst nematodes cause serious yield losses of wheat in Hunaghuai winter wheat growing region in China. Beauveria bassiana 08F04 isolated from the surface of cysts is a promising biological control agent for cereal cyst nematodes. As the colonization capacity is a crucial criteria to assess biocontrol effectiveness for a microbial agent candidate, we aimed to label B. bassiana 08F04 for efficient monitoring of colonization in the soil. The binary pCAM-gfp plasmid containing sgfp and hph was integrated into B. bassiana 08F04 using the Agrobacterium tumefaciens-mediated transformation. The transformation caused a significant change in mycelial and conidial yields, and in extracellular chitinase activity in some transformants. The cultural filtrates of some transformants also decreased acetylcholinesterase activity and the survival of Heterodera filipjevi second-stage juveniles relative to the wild-type strain. One transformant (G10) had a growth rate and biocontrol efficacy similar to the wild-type strain, so it was used for a pilot study of B. bassiana colonization conducted over 13 weeks. Real-time PCR results and CFU counts revealed that the population of G10 increased quickly over the first 3 weeks, then decreased slowly over the following 4 weeks before stabilizing. In addition, the application of wild-type B. bassiana 08F04 and transformant G10 significantly reduced the number of H. filipjevi females in roots by 64.4% and 60.2%, respectively. The results of this study have practical applications for ecological, biological and functional studies of B. bassiana 08F04 and for bionematicide registration.
为筛选防治小麦茎基腐病的高效生防菌株,采用稀释涂布法从小麦茎基腐病病土中分离细菌,以假禾谷镰孢Fusarium pseudograminearum为靶标菌,通过平板对峙培养法和室内盆栽试验筛选优良菌株并测定其田间防治效果,分析优良菌株发酵滤液对假禾谷镰孢的抑菌能力,并结合形态特征、Biolog微生物自动鉴定系统和gyrA序列分析对其进行种类鉴定,以及测定该菌株的抑菌谱.结果 表明,10份供试土样中共分离到98株细菌,对假禾谷镰孢的菌丝生长抑制率超过40.00%的菌株有14株,其中菌株YB-161对假禾谷镰孢的菌丝生长抑制率最高,达63.50%.室内盆栽试验结果显示,菌株YB-161处理小麦的病情指数最低,为31.40,对小麦茎基腐病的防治效果最好,为68.15%,其对小麦也表现出较好的促生作用.田间调查结果显示,菌株YB-161菌液拌种处理对小麦茎基腐病的田间防治效果高于52.35%,并对小麦具有一定的增产作用.菌株YB-161的发酵滤液5倍稀释液和10倍稀释液对假禾谷镰孢的菌落抑制率分别达66.67%和44.33%,该菌株被鉴定为解淀粉芽胞杆菌Bacillus amyloliquefaciens.此外,菌株YB-161对辣椒炭疽病菌Colletotrichum capisci、烟草疫霉病菌Phytophthora parasitica等5种植物病原真菌均有较好的拮抗作用.表明生防菌株YB-161对绿色防控小麦茎基腐病具有良好的开发利用潜能.
由假禾谷镰刀菌Fusarium pseudograminearum引起的小麦茎基腐病是一种重要的土传病害,生产上亟需一种安全有效的防控方法。本研究利用枯草芽胞杆菌YB-05与申嗪霉素复配,并评价该复配制剂对小麦茎基腐病的防治效果。结果显示当申嗪霉素浓度低于500μg/m L时,对YB-05的菌落生长无显著影响;室内盆栽及田间试验表明枯草芽胞杆菌YB-05菌液和1%申嗪霉素悬浮剂按照19:1(vt/vt)复配生物制剂20m L/kg拌种处理发病最轻,均超过了高剂量的单剂处理(枯草芽胞杆菌YB-0530m L/kg拌种处理和1%申嗪霉素2 mL/kg拌种处理)和4.8%适麦丹水悬浮剂2 mL/kg拌种处理,并且具有一定的促生和增产作用。其中,复配制剂室内防治效果为69.8%,在两次的田间调查中防治效果分别为57.8%和45.7%。由此可见,枯草芽胞杆菌与申嗪霉素复配具有协同增效的作用,可以减少化学农药的使用,具有较好的开发利用潜力。