通过导入几丁质酶基因提高巨大芽孢杆菌的生防效果

Plant Diversity ›› 2007, Vol. 29 ›› Issue (06): 666-670.

通过导入几丁质酶基因提高巨大芽孢杆菌的生防效果

张新建¹ , 黄玉杰¹, 杨合同^{1 , 2}, 任艳¹ , 陈凯¹

1 山东省科学院中日友好生物技术研究中心, 山东省应用微生物重点实验室, 山东济南 250014 ;
2 山东理工大学生命科学学院, 山东淄博 255049

收稿日期:2007-06-01 出版日期:2007-12-25 发布日期:2007-12-25
通讯作者: 杨合同

Improving Biocontrol Effect of Bacillus megterium (Bacillaceae) on Plant Disease through Genetic Modification with Chitinase Gene

ZHANG Xin-Jian¹ , HUANG Yu-Jie¹, YANG He-Tong^{1 , 2} , REN Yan¹, CHEN Kai¹

1 Biotechnology Research Center of Shandong Academy of Sciences, Key Laboratory for Applied Microbiology of Shandong Province
Jinan , 250014 , China ; 2 School of Life Sciences, Shandong University of Technology , Zibo 255049 , China

Received:2007-06-01 Online:2007-12-25 Published:2007-12-25
Contact: YANG He-Tong

摘要/Abstract

摘要： 以质粒pMSDChi113 为模板, 经PCR 扩增, 获得了几丁质酶基因1 . 8 kb DNA 片段, 将该基因与大肠杆菌- 芽孢杆菌穿梭质粒pHY300PLK 连接, 获得重组质粒pHYChi113。重组质粒经芽孢杆菌感受态方法转入巨大芽孢杆菌( Bacillus megaterium) Ap25 中, 获得的工程菌株B. megateriums Ap25-chi113 同时具有几丁质酶和内切葡聚糖酶酶活。平板对峙实验结果表明, 工程菌株对病原真菌Rhizoctonia solani , Coniothyrium fuckellii , Fusarium graminearum, Macrophoma kawatsukai , Fusarium oxysporium, Botrytis cinerea , Rhizoctonia cerealis, Colletotrichum gloeosporioides, Bipolarls sorohlulana 的拮抗性能明显提高, 尤其是对Coniothyrium fuckellii 的抑制率相对于野生菌Ap25 提高了13%以上。盆栽防病实验结果表明, 工程菌株显著降低纹枯病( Rhizotonia cerealis) 对小麦( Triticum aestivum L .) 及枯萎病菌( Fusarium oxysporum) 对棉花( Gossypium hirsutum Linn .) 的致病能力, 对这两种病原菌的防效分别是70.34% 和58.51%, 同原始菌株相比, 防效分别提高了27.54%和21.28%。

关键词: 几丁质酶, 芽孢杆菌, 穿梭质粒, 转化

Abstract: Bacillus megaterium Ap25 which can produce endoglucanase was proved as a biocontrol agent of plant disease. The aim of this study is to enhance the biocontrol effect of this strain by introduction of a chitinase gene. A 1. 8 kb DNA fragment containing chitinase gene and SD sequence was amplified with the template pMSDChi113 and was inserted into shuttle vector pHY300PLK to construct a new plasmid, pHYChi113. The recombinant plasmid was transformed into B. megterium, resulting in a new strain named Ap25-chi113. Chitin plate culture and PCR amplification confirmed that Ap25-chi113 contained a functional chitinase gene . Comparing with the wild type Ap25 in dural culture experiment , Ap25- chi113 increased its effect against the pathogenic fungi. Especially, the inhibition percentage against Coniothyrium fuckellii increased about 13%. In pot experiment , Ap25- chi113 also increased the effect on suppression of wheat sheath blight and cotton Fusarium wilt caused by Rhizotonia cerealis and Fusarium oxysporum respectively, comparing with the wild strain.

Key words: Chitinase

中图分类号:

Q 784

张新建1 , 黄玉杰1 , 杨合同1 , 2 , 任艳1 , 陈凯1 . 通过导入几丁质酶基因提高巨大芽孢杆菌的生防效果[J]. Plant Diversity, 2007, 29(06): 666-670.

ZHANG Xin-Jian , HUANG Yu-Jie, YANG He-Tong^, , REN Yan, CHEN Kai. Improving Biocontrol Effect of Bacillus megterium (Bacillaceae) on Plant Disease through Genetic Modification with Chitinase Gene[J]. Plant Diversity, 2007, 29(06): 666-670.

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