连作马铃薯根际干腐病优势病原菌荧光定量PCR快速检测及在根际的动态变化

doi:10.11686/cyxb20130630

草业学报 ›› 2013, Vol. 22 ›› Issue (6): 239-248.DOI: 10.11686/cyxb20130630

连作马铃薯根际干腐病优势病原菌荧光定量PCR快速检测及在根际的动态变化

李瑞琴^1,2,刘星^1,3,邱慧珍^1,3,4*,张文明^1,3,4,张春红^1,3,4,王蒂^3,4,5*,张俊莲^3,4,5,沈其荣⁶

1.甘肃农业大学资源与环境学院,甘肃兰州 730070;
2.甘肃省农业科学院畜草与绿色农业研究所,甘肃兰州 730070;
3.甘肃省干旱生境作物学重点实验室,甘肃兰州730070;
4.甘肃省作物遗传改良与种质创新重点实验室,甘肃兰州730070;
5.甘肃农业大学农学院,甘肃兰州730070;
6.南京农业大学资源与环境学院,江苏南京210095

出版日期:2013-12-20 发布日期:2013-12-20
通讯作者: 李瑞琴(1969-),女,甘肃庆阳人,高级实验师,在读博士。E-mail:liruiqin_524@163.com
*通讯作者。E-mail:hzqiu@gsau.edu.cn,wangd@gsau.edu.cn
作者简介:李瑞琴(1969-),女,甘肃庆阳人,高级实验师,在读博士。
基金资助:
农业部公益性行业(农业)科研专项(201103004),甘肃省科技重大专项(1102NKDM025)和国家科技支撑计划(2012BAD06B03)资助。

Changes in the dominant pathogens causing Fusarium dry rot of potato in rhizospheric soil under continuous potato cropping systems based on real-time quantitative PCR

LI Rui-qin^1,2, LIU Xing^1,3, QIU Hui-zhen^1,3,4, ZHANG Wen-ming^1,3,4, ZHANG Chun-hong^1,3,4, WANG Di^3,4,5, ZHANG Jun-lian^3,4,5, SHEN Qi-rong⁶

1.College of Resources and Environmental Sciences,Gansu Agricultural University, Lanzhou 730070, China;
2.Gansu Academy of Agricultural Sciences, Institute of Green Agricultural and Animal & Pasture Improvement, Lanzhou 730070, China;
3.Gansu Province Key Laboratory of Aridland Crop Science, Lanzhou 730070, China;
4.Gansu Province Key Laboratory of Crop Genetic & Germplasm Enhancement, Lanzhou 730070, China;
5.College of Agronomy, Gansu Agricultural University, Lanzhou 730070, China;
6.College of Resources and Environmental Sciences, Nanjing Agricultural University, Nanjing 210095, China

Online:2013-12-20 Published:2013-12-20

摘要/Abstract

摘要： 为了解根际土壤中土传病害病原菌的积累与连作障碍之间的关系,进一步寻求缓解和克服马铃薯连作障碍土传病害的有效途径,本研究建立了以马铃薯根际土壤为研究对象的干腐病病原菌荧光定量PCR快速检测体系。结果显示,研究建立优化的荧光定量PCR检测方法,对引起马铃薯干腐病的优势病原菌茄病镰孢菌和接骨木镰孢菌进行快速检测和绝对定量,主要优化参数为:上下游引物各0.4 μL(10 mmol/L),DNA模板3 μL,退火温度60℃。连作马铃薯根际土壤茄病镰孢菌和接骨木镰孢菌随连作年限的动态变化趋势均表现为随连作年限的递增呈现上升趋势,其中CP₅的累积量最大,为1.45×10⁴拷贝/g,比CK增加了27.8倍。CP₄、CP₃、CP₂、CP₁根际病原菌累积量分别比对照增加了10.5,16.31,8.32,3.51倍。由此可见,茄病镰孢菌和接骨木镰孢菌的数量均随连作年限的递增呈上升趋势。根际镰刀菌随生育进程的动态变化因病原菌种类而异,茄病镰孢菌以收获期累积量最大,平均为1.2×10⁴拷贝/g,随生育进程的推进呈上升趋势。而接骨木镰刀菌是播前累积量最大,平均为1.55×10⁴ 拷贝/g,随生育进程的推进呈下降趋势。因此,研究区马铃薯根际土壤中镰刀菌的大量积累可能是导致马铃薯连作障碍发生的主要原因之一。

Abstract: The relationship between continuous cropping obstacles and accumulation of pathogens in rhizospheric soil was evaluated, and the mechanisms of continuous potato cropping obstacles explored, using an optimized SYBR Green I dye real time fluorescent quantitative PCR approach. The methods were used to perform quick checks and quantification of Fusarium sambucim and F. solani in soils infested with potato Fusarium dry rot disease and to investigate the dynamic changing trends of F. sambucim and F. solani in rhizospheric soil of continuously cropped potato. The concentration of primers was 0.4 μL (10 mmol/L), of DNA template was 3 μL, and the annealing temperature was 60℃. With an increase in years of continuous cropping potato, the numbers of F. solani and F. sambucim in rhizospheric soil showed an upward trend. The largest amount (1.45×10⁴ copies/g) of F. solani accumulated during the squaring period of five years and it increased 27.8 times compared with the control group (CK). F. solani cumulants CP4, CP3, CP2, CP1 were more than those of RP and increased by 11.4, 35.5, 6.9 and 2.8 times respectively. The largest amount of F. sambucim cumulants (2.21×10⁴ copies/g) was also during the squaring period of five years and compared to the CK they increased 25.0 times. The F. sambucim cumulants CP4, CP3, CP2, CP1 were more than those of the CK and increased by 10.5, 16.31, 8.32 and 3.51 times respectively. Accumulation of rhizospheric Fusarium may be one of the main reasons for the occurrence of obstacles to continuous potato cropping.

中图分类号:

S532
S436.32

李瑞琴,刘星,邱慧珍,张文明,张春红,王蒂,张俊莲,沈其荣. 连作马铃薯根际干腐病优势病原菌荧光定量PCR快速检测及在根际的动态变化[J]. 草业学报, 2013, 22(6): 239-248.

LI Rui-qin, LIU Xing, QIU Hui-zhen, ZHANG Wen-ming, ZHANG Chun-hong, WANG Di, ZHANG Jun-lian, SHEN Qi-rong. Changes in the dominant pathogens causing Fusarium dry rot of potato in rhizospheric soil under continuous potato cropping systems based on real-time quantitative PCR[J]. Acta Prataculturae Sinica, 2013, 22(6): 239-248.

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连作马铃薯根际干腐病优势病原菌荧光定量PCR快速检测及在根际的动态变化

Changes in the dominant pathogens causing Fusarium dry rot of potato in rhizospheric soil under continuous potato cropping systems based on real-time quantitative PCR

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