苹果变色症状伴随相关类病毒的种类、分子特性及与症状相关性分析  

Study on the species,molecular traits and the symptomatic relevance of the viroids in apple fruits showing discoloration symptoms

作　　者：查富蓉 徐勤耕 陈姣 殷玉梦 谌茂秋 王国平[1] 洪霓[1] 王振华 徐文兴[1] ZHA Furong;XU Qingeng;CHEN Jiao;YIN Yumeng;SHEN Maoqiu;WANG Guoping;HONG Ni;WANG Zhenhua;XU Wenxing(College of Plant Science and Technology,Huazhong Agricultural University,Wuhan 430070,China;Technology Center of Wuhan Customs District,Wuhan 430050,China)

机构地区：[1]华中农业大学植物科技学院,武汉430070 [2]武汉海关技术中心,武汉430050

出　　处：《植物病理学报》2023年第5期810-818,共9页Acta Phytopathologica Sinica

基　　金：国家重点研发计划项目(2019YFD1001800);海关总署科研项目(2020HK175)。

摘　　要：长期以来,在苹果果实上较为普遍存在多种变色症状,包括条纹、条斑、斑驳、血色斑点、线纹、花脸、白色圆斑、褪绿环斑、锈斑等症状,疑似为病毒或类病毒引起,但研究较少,与病毒或类病毒侵染的相关性尚不十分清楚,给防治带来困难。本研究收集表现变色症状的26份苹果果实样品,针对苹果锈果类病毒(apple scar skin viroid, ASSVd)、梨泡状溃疡类病毒(pear blister canker viroid, PBCVd)、苹果凹果类病毒(apple dapple fruit viroid, ADFVd)和啤酒花矮化类病毒(hop stunt viroid, HSVd)进行RT-PCR检测。针对检测阳性的类病毒进行克隆、测序和分子特性分析。结果显示,分别有10、5和3份样品呈ASSVd、PBCVd和ADFVd阳性,没有检出HSVd。分析症状与类病毒感染之间的相关性,显示花脸、褪色环斑和部分锈斑症状与ASSVd侵染相关,而白色斑块和白色条斑与ADFVd侵染相关。序列比对分析显示三种类病毒分别在不同区域存在较高的同源性,同源率为38~46%,可分为5个相似的结构区域。进一步采用小RNA深度测序确认表现花脸症状的苹果仅有ASSVd侵染,表明上述症状与ASSVd侵染密切相关。该研究明确了不同变色症状苹果果实内类病毒的种类及其分子特性,该研究可望为果园内类病毒快速诊断提供参考,并为采用现代分子技术防治该类病害提供依据。Diverse discoloration symptoms(streak,mottle,blood-colored spot,short-line stripe,dapple,white spot,ringspot,scar,chlorosis,etc.)often appear on apple fruits,which are suspected to be the results of viroid infection.The relevance of these diverse symptoms with viroid is underdetermined,and it brings some difficulty for the control of viroids.A total of 26 apple fruit samples showing different discoloration symptoms were collected,and subjected to RT-PCR analysis for the infection of diverse viroids that have previously been reported infecting apple plants,namely,apple scar skin viroid(ASSVd),pear blister canker viroid(PBCVd),apple dapple fruit viroid(ADFVd),and hop stunt viroid(HSVd).The target bands of viroids detected from the apple samples were cloned,sequenced,and their molecular traits were analyzed.The results showed that ten,five,and three samples were positive for ASSVd,PBCVd and ADFVd,respectively,while no samples were positive for HSVd.Sequence alignment reveals the identities ranging from 38 to 46%among them,with some obviously homology regions.Analysis of the correlation between the symptoms and the viroid infections showed that the symptoms of dapple,chlorotic ringspots and some special scar were related to ASSVd infection,and white round spots were related to ADFVd,whereas the symptoms such as mottle,blood-colored spot,short streak,and the remaining scar symptoms were not related to the infection of viroids.Further deep sequencing of small RNAs(sRNAs)was performed on YT-1 sample,and it confirms that the symptoms including dapple and chlorotic ringspots are related to ASSVd instead of other viroids.This study clarified the correlation between the various discoloration symptoms of apple fruits and the infection of viroids,provides basic information for prevention and controlling of these diseases showing diverse discoloration symptoms,and it will provide useful information for diagnose and control of the resulting diseases.

关 键 词：苹果 变色 花脸 类病毒 苹果锈果类病毒 苹果凹果类病毒 

分 类 号：S432.41[农业科学—植物病理学]