尤力克柠檬上一种新病害的生物学特性及RT-PCR检测  

Biological characterization and RT-PCR detection of a new disease of Eureka lemon

作　　者：陈洪明[1,2] 王雪峰[1,2] 周彦[1,2] 周常勇[1,2] 郭俊[3] 李中安[1,2] 

机构地区：[1]西南大学 [2]中国农业科学院柑橘研究所,重庆400712 [3]云南省农业科学院热带亚热带经济作物研究所,保山678025

出　　处：《植物保护学报》2015年第4期557-563,共7页Journal of Plant Protection

基　　金：国家公益性行业(农业)科研专项(201203076);重庆市自然科学基金(cstc2011jj A80025);中央高校基本科研业务费专项(XDJK2012C101;XDJK2014A001)

摘　　要：近年我国云南瑞丽市的一果园尤里克柠檬上出现一种引起叶片皱缩、反卷或呈船形,叶背水浸状、侧脉脉明、黄化的柑橘新病害。为明确该病害的病原及其生物学特性,通过田间观察、病样接种、电镜观察,并基于柑橘黄脉病毒（Citrus yellow vein clearing virus,CYVCV）外壳蛋白基因设计引物对病样进行RT-PCR检测。结果表明,该病是一种嫁接传染性病害,能侵染柠檬和酸橙品种且症状明显,并能摩擦接种到辣椒和豇豆等草本植物。该病发病最适温度为18~24℃。春梢嫩叶症状表现明显,夏梢和秋梢基本不表现症状,而晚秋梢也表现出较春梢稍弱的典型症状。引致该病的病毒为（13~15）nm×（400~1 000）nm大小的联合丝状病毒。RT-PCR扩增产物为614 bp,且其与CYVCV外壳蛋白序列相似性达97%以上,表明该病害是由CYVCV引起的柑橘黄脉病,并初步建立了该病的RT-PCR检测技术。A new disease epidemic on Eureka lemon（ Citrus limon（ L.） Burm. f.） was discovered in an orchard of Ruili City in Yunnan Province. The disease caused typical symptoms,including leave crinkle or curl-up,lateral vein yellowing and clearing against light,and water-soaked appearance were also seen on the adaxial surface of leaves. To determine the causal agent and biological features of the disease,comprehensive detection were carried out,including field survey,indicator plant inoculation,transmission electron microscopy observation and RT-PCR assay by using the primers designed from coat protein gene of Citrus yellow vein clearing virus（ CYVCV） genome. The results showed that this disease could be transmissible in lemon and sour orange（ C. aurantium） by grafting-inoculation, and in capsicum（ Capsicum frutescens） and cowpea（ Vigna unguiculata） by rubbing-inoculation. The optimum temperature for this disease development was 18- 24℃. Symptoms were obviously observed on young leaves of spring shoots,but less obvious on summer and autumn shoots,and reappeared on late autumnshoots. Filamentous virus particles of about（ 13- 15） nm ×（ 400- 1 000） nm in width and length were measured with transmission electron microscopy. RT-PCR produced an expected band of 614 bp from all symptomatic samples and nucleotide sequences shared 97% similarity with the known region of CYVCV reference genome. Based on these assays,the new disease on lemon might be caused by CYVCV,and the RT-PCR method was reliably established for detection of this disease.

关 键 词：柠檬 柑橘黄脉病 生物学特性 嫁接传播 RT-PCR 

分 类 号：S436.669[农业科学—农业昆虫与害虫防治]