摘要
菰黑粉菌(Ustilago esculenta)是一种专性寄生植物茭白(Zizania latifolia)的活体营养型真菌,菌丝态菰黑粉菌(MT型)侵染茭白后可以使植株茎部膨大形成可食用肉质茎,即茭白。田间种植中还存在冬孢子态菰黑粉菌(T型),其侵染形成的膨大肉质茎内布满黑色冬孢子,无法食用,称作灰茭。为及早发现并去除灰茭植株,本研究对比了T型与MT型菰黑粉菌内转录间隔区序列(ITS),利用ITS上的单核苷酸多态性位点设计TaqMan探针与引物。本研究筛选得到了具有良好特异性的TaqMan探针及引物,优化了反应体系中的引物浓度与探针浓度,建立并优化了菰黑粉菌分型鉴定方法。该方法不受植物基因组DNA等基质影响,且菰黑粉菌基因组浓度在0.5~2000 ng·L^(-1)范围内时,结果具有良好的线性关系,判定系数(R^(2))>0.99。通过该方法对植物中T型和MT型菰黑粉菌分型定量,可以在茭白种植早期发现将结灰茭的植株,为灰茭预防及栽培措施改良提供新的参考。
Ustilago esculenta is a biotrophic fungus that parasitizes Zizania latifolia obligately.Being infected by mycelia-teliospore U.esculenta(MT-type),the stem of Z.latifolia will swell into an edible fleshy one,namely,jiaobai.In field cultivation,teliospore U.esculenta(T-type)is also existed,and it can also infect Z.latifolia and induce the expansion of the stem.However,such stem filled with black teliospore,namely,grey jiaobai,is inedible.To remove plants infected by T-type U.esculenta at an early stage,the internal transcribed spacer sequence(ITS)of T-type and MT-type U.esculenta were compared,and we designed Taq Man probes and primers using the single nucleotide polymorphism loci in ITS.In this study,the Taq Man probe and primers with good specificity were screened,to establish and optimize their concentration in the reaction system.The method will not be affected by plant genomic DNA or other substrates in the matrix.The results show a good linear relationship ranging from 0.5 to 2000 ng·L^(-1)genomic DNA,with coefficient of determination(R^(2))0.99.By quantifying and genotyping T-type and MT-type U.esculenta intra-plant,the plants which would grow into inedible grey jiaobai could be found at an early stage.It provides a new way for the prevention of grey jiaobai and the improvement of cultivation measures.
作者
李士玉
杨梦飞
夏文强
张雅芬
汤近天
崔海峰
叶子弘
李怡鹏
张尚法
张中进
王书卿
LI Shiyu;YANG Mengfei;XIA Wenqiang;ZHANG Yafen;TANG Jintian;CUI Haifeng;YE Zihong;LI Yipeng;ZHANG Shangfa;ZHANG zhongjin;WANG Shuqing(Zhejiang Provincial Key Laboratory of Biometeorology and Inspection&Quarantine,College of Life Sciences,China Jiliang University,Hangzhou 310018,China;Zhejiang Provincial Key Laboratory of Characteristic Aquatic Vegetable Breeding and Cultivation,Jinhua Academy of Agricultural Sciences(Zhejiang Institute of Agricultural Machinery),Jinhua,Zhejiang 321000,China)
出处
《植物生理学报》
CAS
CSCD
北大核心
2023年第8期1616-1625,共10页
Plant Physiology Journal
基金
国家自然科学基金联合基金(U20A2043)
中国计量大学基本科研业务费项目(2021YW64)
浙江省农业(蔬菜新品种选育)新品种选育重大科技专项(2021C02065)。
关键词
茭白
灰茭
菰黑粉菌
实时荧光定量PCR
Zizania latifolia
grey jiaobai
Ustilago esculenta
quantitative real-time PCR
