Shrestha Bishnu K, Karki Hari Sharan, Groth Donald E, Jungkhun Nootjarin, Ham Jong Hyun
Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge, Louisiana, 70803, United States of America.
Rice Research Station, Louisiana State University Agricultural Center, Rayne, Louisiana, 70578, United States of America.
PLoS One. 2016 Jan 14;11(1):e0146764. doi: 10.1371/journal.pone.0146764. eCollection 2016.
Potential biological control agents for two major rice diseases, sheath blight and bacterial panicle blight, were isolated from rice plants in this study. Rice-associated bacteria (RABs) isolated from rice plants grown in the field were tested for their antagonistic activities against the rice pathogens, Rhizoctonia solani and Burkholderia glumae, which cause sheath blight and bacterial panicle blight, respectively. Twenty-nine RABs were initially screened based on their antagonistic activities against both R. solani and B. glumae. In follow-up retests, 26 RABs of the 29 RABs were confirmed to have antimicrobial activities, but the rest three RABs did not reproduce any observable antagonistic activity against R. solani or B. glumae. According to16S rDNA sequence identity, 12 of the 26 antagonistic RABs were closest to Bacillus amyloliquefaciens, while seven RABs were to B. methylotrophicus and B, subtilis, respectively. The 16S rDNA sequences of the three non-antagonistic RABs were closest to Lysinibacillus sphaericus (RAB1 and RAB12) and Lysinibacillus macroides (RAB5). The five selected RABs showing highest antimicrobial activities (RAB6, RAB9, RAB16, RAB17S, and RAB18) were closest to B. amyloliquefaciens in DNA sequence of 16S rDNA and gyrB, but to B. subtilis in that of recA. These RABs were observed to inhibit the sclerotial germination of R. solani on potato dextrose agar and the lesion development on detached rice leaves by artificial inoculation of R. solani. These antagonistic RABs also significantly suppressed the disease development of sheath blight and bacterial panicle blight in a field condition, suggesting that they can be potential biological control agents for these rice diseases. However, these antagonistic RABs showed diminished disease suppression activities in the repeated field trial conducted in the following year probably due to their reduced antagonistic activities to the pathogens during the long-term storage in -70C, suggesting that development of proper storage methods to maintain antagonistic activity is as crucial as identification of new biological control agents.
本研究从水稻植株中分离出了两种主要水稻病害(纹枯病和细菌性穗枯病)的潜在生物防治剂。对从田间种植的水稻植株中分离出的水稻相关细菌(RABs)进行了测试,以检测它们对分别引起纹枯病和细菌性穗枯病的水稻病原菌立枯丝核菌和颖壳伯克霍尔德菌的拮抗活性。最初基于对立枯丝核菌和颖壳伯克霍尔德菌的拮抗活性筛选出了29株RABs。在后续复测中,29株RABs中的26株被证实具有抗菌活性,但其余3株RABs对立枯丝核菌或颖壳伯克霍尔德菌未表现出任何可观察到的拮抗活性。根据16S rDNA序列同一性,26株拮抗RABs中的12株与解淀粉芽孢杆菌亲缘关系最近,而7株RABs分别与甲基营养型芽孢杆菌和枯草芽孢杆菌亲缘关系最近。3株非拮抗RABs的16S rDNA序列与球形赖氨酸芽孢杆菌(RAB1和RAB12)和巨大赖氨酸芽孢杆菌(RAB5)亲缘关系最近。挑选出的5株抗菌活性最高的RABs(RAB6、RAB9、RAB16、RAB17S和RAB18)在16S rDNA和gyrB的DNA序列上与解淀粉芽孢杆菌亲缘关系最近,但在recA的DNA序列上与枯草芽孢杆菌亲缘关系最近。观察到这些RABs能抑制立枯丝核菌在马铃薯葡萄糖琼脂上的菌核萌发以及通过人工接种立枯丝核菌在离体水稻叶片上的病斑发展。这些拮抗RABs在田间条件下也显著抑制了纹枯病和细菌性穗枯病的病害发展,表明它们可能是这些水稻病害的潜在生物防治剂。然而,在次年进行的重复田间试验中,这些拮抗RABs的病害抑制活性有所下降,这可能是由于它们在-70°C长期保存期间对病原菌的拮抗活性降低,这表明开发合适的保存方法以维持拮抗活性与鉴定新的生物防治剂同样重要。