摘要
目的:该文从多方面进行分析、比较,及时、准确地鉴别广佛手黄龙病,以便于及时确定病害、防控病情。方法:通过性状分析,逆转录-聚合酶链式反应(RT-PCR),酶切反应及实时荧光定量PCR(Real-time PCR)检测多种手段相结合,对广佛手黄龙病进行鉴定。结果:从性状上来看,感病广佛手有典型的叶片斑驳型黄化、果相对较小,甚至畸形的特征,但并没有发现"红鼻子果"现象,这些均可以作为田间初步判别广佛手黄龙病的依据;RT-PCR检测及酶切反应结果表明,引物为OI1/OI2c时,广佛手病株有特异性条带1160 bp,且能被Xba I酶切成520 bp和640 bp,这与其他被黄龙病侵染的柑橘属植物的检测结果一致;Real-time PCR检测结果显示,被黄龙病侵染的广佛手叶片有扩增曲线和熔解曲线,熔链温度为82℃,且Ct值在24.6~28.2,而广佛手正常株没有扩增现象出现。结论:性状分析可以在田间初步判别黄龙病,但是具有一定的主观性,RTPCR及Real-time PCR检测能进一步确定广佛手黄龙病,且q PCR检测更加灵敏,还可以定量。通过性状分析以及分子鉴定的结合,能够更加及时、准确地确定广佛手黄龙病。
Objective:To analyze and compare different samples in many aspects to identify Citrus medica var.sarcodactylis infected with Huanglongbing(HLB)timely and accurately,in order to prevent and control the disease in time.Method:HLB was identified through character analysis,reverse transcriptionpolymerase chain reaction(RT-PCR),enzyme digestion reaction and Real-time PCR.Result:In terms of characters,there were typically variegated yellow leaves and relatively small fruit,even with deformity but without"red nose fruit"among C.medica var.sarcodactylis infected with HLB.All of these can be used as the basis for the preliminary identification of HLB in the fields.According to the RT-PCR test results and enzyme digestion reaction,when the primer was OI1/OI2 c,there was specific band of 1160 bp,which could be cut into 520 bp and 640 bp by Xba I enzyme.These results were consistent with the characters of other citrus plants infected with HLB.According to the Real-time PCR detection results,C.medica var.sarcodactylis infected with HLB had amplification curves and dissolved peaks,with the melting temperature was 82℃and Ctbetween 24.6 to 28.2,while the normal plants were not amplified.Conclusion:Character analysis can be used to roughly distinguish HLB in the fields,but with a certain subjectivity.RT-PCR or Real-time PCR can be used to identify C.medica var.sarcodactylis infected with HLB in a timely and accurate manner,and qPCR detection is more sensitive and quantitative.Through the combination of character analysis and molecular identification,C.medica var.sarcodactylis infected with HLB can be determined more timely and accurately.
作者
袁蒙
潘利明
周良云
罗碧
王浩
杨全
YUAN Meng;PAN Li-ming;ZHOU Liang-yun;LUO Bi;WANG Hao;YANG Quan(Guangdong Provincial Research Center on Good Agricultural Practice&Comprehensive Agricultural Development Engineering Technology of Cantonese Medicinal Materials,Guangzhou Comprehensive Experimental Station of China Chinese Materia Medica Industry Technology System,Key Laboratory for Production&Development of Cantonese Medicinal Materials Under State Administration of Traditional Chinese Medicine(TCM),School of TCM,Guangdong Pharmaceutical University,Guangzhou 510006,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第5期117-122,共6页
Chinese Journal of Experimental Traditional Medical Formulae
基金
国家重点研发计划项目(2017YFC1700704)
广东省中医药局中药资源普查专项(20163008)
2017年广东省岭南中药材保护资金专项(粤财社[2017]60号).
关键词
广佛手
黄龙病
叶片
性状分析
田间判别
Citrus medica var.sarcodactylis
Huanglongbing(HLB)
leaves
character analysis
distinguish in field
作者简介
第一作者:袁蒙,在读硕士,从事中药资源开发与品质评价研究,Tel:020-39252353,E-mail:964485970@qq.com;通信作者:杨全,博士,教授,从事中药资源开发与品质评价研究,Tel:020-39252353,E-mail:yangquan7208@vip.163.com
