白僵菌施加对水稻三种抗氧化酶活力及叶际微生物多样性的影响

Abstract

PDF HTML阅读 XML下载 导出引用 引用提醒 白僵菌施加对水稻三种抗氧化酶活力及叶际微生物多样性的影响 DOI: 10.5846/stxb201303050350 作者: 作者单位: 南京师范大学生命科学学院,南京师范大学生命科学学院,南京师范大学生命科学学院,南京师范大学生命科学学院,南京师范大学生命科学学院,环保部南京科学研究所,南京大学生命科学学院,南京师范大学生命科学学院 作者简介: 通讯作者: 中图分类号: 基金项目: 环境保护部环保公益性行业科研专项(201009023) Effects of Beauveria bassiana on paddy antioxidant enzymes activities and phyllosphere microbial diversity Author: Affiliation: Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources,Jiangsu Key Laboratory for Microbes and Functional Genomics,College of Life Science,Nanjing Normal University;China;Nanjing Institute of Environmental Science,Ministry of Environment Protection;China;School of Life Science,Nanjing University;China,Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources,Jiangsu Key Laboratory for Microbes and Functional Genomics,College of Life Science,Nanjing Normal University,Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources,Jiangsu Key Laboratory for Microbes and Functional Genomics,College of Life Science,Nanjing Normal University,,,,,Jiangsu Engineering and Technology Research Center for Industrialization of Microbial Resources,Jiangsu Key Laboratory for Microbes and Functional Genomics,College of Life Science,Nanjing Normal University Fund Project: 摘要 | 图/表 | 访问统计 | 参考文献 | 相似文献 | 引证文献 | 资源附件 | 文章评论 摘要:为了考察白僵菌使用后的生态安全性,研究了白僵菌施加对水稻3种保护酶活力及叶际微生物多样性的影响。使用荧光定量PCR对喷洒白僵菌孢子悬液的水稻叶际提取总DNA并进行荧光定量PCR扩增,发现白僵菌可以在水稻叶际残留达30 d之久,并且初始喷施浓度越大,衰减速率越高。与施加化学农药相比,施加白僵菌没有对水稻叶片的3种抗氧化酶活力带来不利影响。白僵菌处理组SOD、POD活力在第10天时高于对照组20.38%、8.65%,而CAT活力在第30天时最高高于对照组33.67%,在第30天时,化学农药导致CAT活力相比较对照组下降了42.71%。使用DGGE分析表明白僵菌对水稻叶际细菌、真菌微生物群落结构影响较小,并且白僵菌处理组微生物群落结构相似度、香农指数及条带数均较高。结果表明白僵菌是一种环境友好型的微生物农药。 Abstract:Beauveria bassiana is a well-known entomopathogenic fungi with worldwide distribution that can be used as a microbial pesticide. To date, studies of B. bassiana have focused on its insecticidal mechanism, symbiosis with plants and antagonism of plant pathogens; however, few studies of its influence on the microecological phyllosphere and at physiological level of the plant have been conducted. To investigate the ecological security of B. bassiana in the paddy phyllosphere, we evaluated the effects of different concentrations of B. bassiana spore suspensions and a chemical pesticide (acephate) on paddy phyllosphere microbial flora and protective enzyme activities by a potted trial. B. bassiana were then induced to express the egfp gene (green fluorescent protein gene) through transformation with a vector, after which real-time polymerase chain reaction (PCR) was used to quantify the organisms and measure their dynamics in a paddy phyllosphere. We also evaluated the effects of different concentrations of B. bassiana suspensions on the phyllosphere microflora using DGGE. To accomplish this, B. bassiana specific DNA primers were designed based on the green fluorescent protein sequence marked B. bassiana. Amplification of B. bassiana DNA using the eGPF-F1/eGFP-R1 primers yielded a single 289bp-long product with a detection limit of 10fg/μL of B. bassiana genomic DNA. The pot experiments, which were conducted in the botanical garden of Nanjing Normal University, consisted of the following seven treatments: sterile water applied as a control (CK), inoculation with the larva of Chilo suppressalis (A), application of the B. bassiana spore suspension at 7.5×104spores/ml (B), 7.5×105 spores/ml (C), 7.5×106 spores/ml (D), or 7.5×107 spores/ml (E), application of acephate emulsifiable concentrates (F). Each treatment group was covered with gauze (3 m × 0.7 m × 1.5 m) after treatment. Fluorescence quantitative PCR analysis of the rice phyllosphere DNA revealed that B. bassiana was maintained for at least 30 days on the leaves of paddy plants after application, but when a higher initial dosage of the B. bassiana was applied, the B. bassiana population on the phyllosphere decreased more rapidly. When compared with chemical pesticide, B. bassiana did not significantly affect the antioxidant activity of three enzymes in paddy leaves. Specifically, the enzymatic activities of SOD and POD in B. bassiana treatment groups were 20.38 and 8.65% higher, respectively, than those in the CK group on the tenth day, while the activity of CAT was 33.67% higher than that of the CK group on day 30. However, the enzymatic activity of CAT in the F group was 42.71% lower than that of the CK group on day 10. DGGE cluster analysis showed that B. bassiana did not significantly influence the bacterial or fungal community structures on the paddy phyllosphere, and the microbial community structure similarity, Shannon index and band number in the B. bassiana treated group were higher than in the negative control group. These results indicated that B. bassiana is an environmental friendly microbial pesticide. Future studies of B. bassiana should focus on its effects on other endophytes in plants. Studies to evaluate the role of B. bassiana could lead to a new paradigm on its successful use in biological control programs. 参考文献 相似文献 引证文献