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用于防治小麦赤霉病的内生芽孢杆菌多样性及探索

Diversity and Exploration of Endophytic Bacilli for the Management of Head Scab () of Wheat.

作者信息

Kaul Noyonika, Kashyap Prem Lal, Kumar Sudheer, Singh Deepti, Singh Gyanendra Pratap

机构信息

Division of Crop Protection, ICAR-Indian Institute of Wheat and Barley Research, Karnal 132001, Haryana, India.

Amity Institute of Microbial Technologies, Amity University, Karnal 132001, Haryana, India.

出版信息

Pathogens. 2022 Sep 23;11(10):1088. doi: 10.3390/pathogens11101088.

Abstract

Fusarium graminearum causing head scab (HS) or head blight (HB) disease in wheat is one of the nasty fungi reported to cause significant grain quality and yield loss. Biological control using endophytic bacteria has emerged as a prospective option for containing fungal diseases in an environmentally benevolent, durable, and sustainable manner. In this regard, 112 endophytic bacilli were isolated from the anthesis stage (Zadok’s growth stage 65) from five different wheat genotypes with an aim to identify prospective antagonistic strains against F. graminearum. The molecular identity of the strains was confirmed by matching 16S rRNA sequences of bacterial strains with the gene sequences of type strains available in the National Center for Biotechnology Information database and reported 38 different species of Bacillus in all the five wheat cultivars. Further, it has been observed that only fourteen strains (B. clarus NOK09, B. mojavensis NOK16, B. subtilis NOK33, B. rugosus NOK47, B. mojavensis NOK52, B. clarus NOK59, B. coahuilensis NOK72, B. cabrialesii NOK78, B. cabrialesii NOK82, B. rugosus NOK85, B. amyloliquefaciens NOK89, B. australimaris NOK95, B. pumilus NOK103, and B. amyloliquefaciens NOK109) displayed in-vitro antagonistic effect against Fusarium graminearum fungus. Furthermore, the three endophytic Bacillus strains showing the strongest antagonistic effect (>70% of growth inhibition of fungal mycelium) under in-vitro antagonistic assay were selected for field experiments. In a two-year consecutive field study, a combination of three strains (B. clarus NOK09 + B. subtilis NOK33 + B. amyloliquefaciens NOK109) displayed a remarkable reduction in HS disease index by 81.47% and 77.85%, respectively. Polymerase chain reaction assay detected three genes (ituD, bmyC, and srfA) involved in antibiotic biosynthesis pathways. Additional attributes such as potassium solubilization, siderophore release, and hydrolytic enzyme (protease, lipase, amylase, chitinase, and pectinase) synthesis have been observed in these strains. Overall, the present study was successful in profiling endophytic bacilli and selecting the combination of effective antagonistic endophytic Bacillus strains that could be the best alternative for the sustainable and ecological sound management of HS disease in wheat under field conditions.

摘要

引起小麦赤霉病(HS)或赤霉枯萎病(HB)的禾谷镰刀菌是一种有害真菌,据报道会导致严重的谷物品质和产量损失。利用内生细菌进行生物防治已成为一种以环境友好、持久和可持续的方式控制真菌病害的潜在选择。在这方面,从五个不同小麦基因型的开花期(扎多克生长阶段65)分离出112株内生芽孢杆菌,旨在鉴定对禾谷镰刀菌有潜在拮抗作用的菌株。通过将细菌菌株的16S rRNA序列与美国国立生物技术信息中心数据库中可用的模式菌株的基因序列进行匹配,确认了菌株的分子身份,并报道了所有五个小麦品种中共有38种不同的芽孢杆菌。此外,观察到只有十四株菌株(克拉鲁氏芽孢杆菌NOK09、莫哈韦芽孢杆菌NOK16、枯草芽孢杆菌NOK33、粗糙芽孢杆菌NOK47、莫哈韦芽孢杆菌NOK52、克拉鲁氏芽孢杆菌NOK59、科阿韦拉芽孢杆菌NOK72、卡布里亚莱斯芽孢杆菌NOK78、卡布里亚莱斯芽孢杆菌NOK82、粗糙芽孢杆菌NOK85、解淀粉芽孢杆菌NOK89、澳大利亚芽孢杆菌NOK95、短小芽孢杆菌NOK103和解淀粉芽孢杆菌NOK109)对禾谷镰刀菌显示出体外拮抗作用。此外,选择了在体外拮抗试验中表现出最强拮抗作用(真菌菌丝体生长抑制率>70%)的三株内生芽孢杆菌菌株进行田间试验。在一项为期两年的连续田间研究中,三株菌株的组合(克拉鲁氏芽孢杆菌NOK09 + 枯草芽孢杆菌NOK33 + 解淀粉芽孢杆菌NOK109)分别使赤霉病病害指数显著降低了81.47%和77.85%。聚合酶链反应分析检测到参与抗生素生物合成途径的三个基因(ituD、bmyC和srfA)。在这些菌株中还观察到了其他特性,如钾溶解、铁载体释放和水解酶(蛋白酶、脂肪酶、淀粉酶、几丁质酶和果胶酶)合成。总体而言,本研究成功地对内生芽孢杆菌进行了分析,并选择了有效的拮抗内生芽孢杆菌菌株组合,这可能是田间条件下可持续和生态合理管理小麦赤霉病的最佳替代方案。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bcd2/9609341/32fc26854668/pathogens-11-01088-g001.jpg

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