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浙江大学学报（农业与生命科学版）
Journal of Zhejiang University (Agriculture and Life Sciences)  2015, Vol. 41 Issue (03): 277-284    DOI: 10.3785/j.issn.1008-9209.2014.10.251
Biological sciences & biotechnology     
Detection of real-time fluorescence quantitative polymerase chain reaction for colonization of Bacillus amyloliquefaciens and Trametes versicolor in Juglans sigllata Dode rhizosphere soil
Zheng Lei, Zhang Jing, Peng Yan, Ma Wenjian, Zhu Tianhui*
(College of  Forestry, Sichuan Agricultural University, Ya’an 625014, Sichuan, China)
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Abstract  Juglans sigllata Dode,  is widely planted in Liangshan Prefecture of southwest China for its nuts and wood. Liangshan Prefecture is a major traditional growing area of J. sigllata and has unique advantages for walnut industrial development because of its good soil, climate, and availability of water. Currently there are 2.7 million hectares of walnut, contributing important incomes for farmers. In April 2013, numerous J. sigllata were found to be infected with root rot in the Muli  County of Liangshan Prefecture. Symptoms included dried leaves, dead branches, and even death.  According to the cultural characteristics, the fungus was primarily identified as Fusarium solani and it  could cause the root rot on many industrial crops. Fusarium  solani could  be significantly antagonized by Bacillus amyloliquefaciens and Trametes versicolor. In order to control the plant diseases caused by F. solani, B. amyloliquefaciens and T. versicolor  were chosen for testing the efficiency of the colonization in rhizosphere soil of J. sigllata. With the development of real-time fluorescence quantitative polymerase chain reaction (PCR) technology, more attention was paid on this issue. It was highly efficient, sensitive and specific technology and had been widely used in various fields of scientific research. For the research of the root rot of J. sigllata, the dynamic changes of two kinds of biocontrol strains and the pathogen in rhizosphere soil were studied through the method of real-time fluorescence quantitative PCR. The results showed that biocontrol strains multiplied rapidly after inoculation, and the relative content of B. amyloliquefaciens reached the peak at the 20th day. Then, the relative content of B. amyloliquefaciens was declined, and kept stable after the 40th day. But the time when the relative content of T. versicolor showed the growth peak and stabilization at each stage was delayed for 10 d as compared with B. amyloliquefaciens. After 60 days, the relative contents of F. solani in rhizosphere soil inoculated with B. amyloliquefaciens and T. versicolor were reduced significantly when the relative content of biocontrol strains reached the peak. The difference in the content of B. amyloliquefaciens was significant between the treatment with or without inoculating B. amyloliquefaciens, while more distinctions of the content of T. versicolor between the treatment with or without inoculating T. versicolor. When the colonization was stabilized, the content of B. amyloliquefaciens had no significant difference compared with the control, while the content of T. versicolor was significant between the two treatments. All above indicate  that the biocontrol efficiency and the colonization ability of B. amyloliquefaciens are   superior than T. versicolor. This study can  provide real-time monitoring of diseases caused by F. solani and support the detection of the colonization of B. amyloliquefaciens and T. versicolor in rhizosphere soil.

Published: 20 May 2015
CLC:  Q 936  
  S 763.1  
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Zheng Lei
Zhang Jing
Peng Yan
Ma Wenjian
Zhu Tianhui
Cite this article:

Zheng Lei, Zhang Jing, Peng Yan, Ma Wenjian, Zhu Tianhui. Detection of real-time fluorescence quantitative polymerase chain reaction for colonization of Bacillus amyloliquefaciens and Trametes versicolor in Juglans sigllata Dode rhizosphere soil. Journal of Zhejiang University (Agriculture and Life Sciences), 2015, 41(03): 277-284.

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http://www.zjujournals.com/agr/10.3785/j.issn.1008-9209.2014.10.251     OR     http://www.zjujournals.com/agr/Y2015/V41/I03/277


实时荧光定量聚合酶链反应检测核桃根际土壤生防菌Bacillus amyloliquefaciens和Trametes versicolor的定殖

以铁核桃根腐病为研究对象,运用实时荧光定量聚合酶链反应(polymerase chain reaction,PCR)扩增技术,分别对铁核桃根际土壤中解淀粉芽孢杆菌(Bacillus amyloliquefaciens)和白腐菌(Trametes versicolor)2种生防菌及腐皮镰刀菌(Fusarium solani)病原菌的数量动态变化进行研究。结果表明:2种生防菌在接种后短期内都能够迅速生长繁殖,B. amyloliquefaciens相对含量在第20天达到峰值,20 d后呈下降趋势,40 d后趋于稳定,而T. versicolor在各时期的相对含量变化均较前者延迟10 d;60 d后,在施入B. amyloliquefaciens和T. versicolor的根际土壤中F. solani的相对含量极显著降低;在生防菌的相对含量达到峰值时,B. amyloliquefaciens与其对照组相比差异有统计学意义(P＜0.05),T. versicolor则差异有高度统计学意义(P＜0.01);定殖稳定后,B. amyloliquefaciens与其对照组相比差异无统计学意义(P＞0.05),T. versicolor对照组与其相比差异有统计学意义(P＜0.05),2种生防菌在铁核桃根际土壤中均有很强的生防效果和定殖能力,生防效果为B. amyloliquefaciens>T. versicolor,定殖能力为B. amyloliquefaciens>T. versicolor。该研究为由腐皮镰刀菌引起的病害实时监测提供了重要参考,同时为2种生防菌在土壤中的定殖检测提供了有力的技术支持。
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