我国水稻黑条矮缩病和玉米粗缩病研究进展

上世纪60年代初我国华东和华北地区分别发现了水稻黑条矮缩病和玉米粗缩病,后来都分别发生了2次大流行,同时都以病害防治为目标分别开展了研究,探明了当地病害发生规律,提出了相应的防治方法,获得了不同程度的防病效果.80年代以来,水稻黑条矮缩病的发生报道仅限于华东地区局部地市,玉米粗缩病的发生涉及华北、东北、西北、西南和华中地区13个省市.根据各地对两病病原形态、寄主及症状、介体昆虫及传病特性等方面相似性的报道,提出了我国玉米粗缩病与水稻黑条矮缩病病原异同性问题.经近10年来对两病用生物学和分子生物学方法进行比较鉴定,证明我国玉米粗缩病和水稻黑条矮缩病病原同属水稻黑条矮缩病毒(RBSDV).同时基本探明了水稻黑条矮缩病在浙江杂交稻区和华北玉米区的再次流行成灾的原因,提出了相应的防治措施,并有效地控制了病害的流行危害.     

水稻黑条矮缩病和玉米粗缩病的发生特点及防治策略

本文着重介绍水稻黑条矮缩病的主要发生危害特点及防治策略,以加深对新列入湖北省植物检疫补充对象的了解与认识,掌握“两病”的主要传播途径。强化植物措施,对保护区域性农业生产安全具有重要实践与指导意义。     

玉米粗缩病和水稻黑条矮缩病的发生及防治对策

本文根据玉米粗缩病和水稻黑条矮缩病近几年来的发生特点和为害情况 ,具体阐述这两种病害的发病症状、发生规律、病原生物学、发生影响因素、合理的防治措施 ,为今后更好地识别和防治这两种病害提供了理论基础     

南方水稻黑条矮缩病毒和水稻黑条矮缩病毒的单抗制备及其检测应用

 用与牛血清白蛋白偶联的南方水稻黑条矮缩病毒（Southern rice black-streaked dwarf virus,SRBSDV）衣壳蛋白的C端12个氨基酸多肽为抗原免疫BALB/c小鼠,经细胞融合、筛选、克隆,获得2株能稳定传代并分泌抗SRBSDV和水稻黑条矮缩病毒（Rice black-streaked dwarf virus,RBSDV）单克隆抗体（MAb）的杂交瘤细胞株3F1、5G1。3F1、5G1单克隆抗体腹水间接ELISA效价达10^-6,抗体类型及亚类均为IgG1, kappa链。 Western blot分析表明,2株单克隆抗体均与SRBSDV和RBSDV的外壳蛋白亚基有特异反应。利用单克隆抗体3F1建立的dot-ELISA检测方法能准确、特异、灵敏地检测田间稻飞虱及水稻样品中的SRBSDV和RBSDV。SRBSDV和RBSDV单克隆抗体的制备及检测方法的建立为水稻黑条矮缩病的诊断、预测预报及科学防控提供了技术支撑。     

水稻黑条矮缩病和玉米粗缩病的发生特点及防治策略

水稻黑条矮缩病和玉米粗缩病都是病毒病害。“两病”的重要感病期平均为营养生长至生殖生长的过渡期,危害盛期也都在生殖生长旺期。受害作物感病愈早,危害愈重。“两病”都能致受害作物矮化,生长畸形,阻碍正常生长,严重影响产量和品质。     

水稻黑条矮缩病毒江苏玉米分离物全基因组序列及进化分析

 利用RT-PCR从江苏玉米上克隆水稻黑条矮缩病毒(RBSDV)的全基因组,其基因组10个片段核苷酸数目和蛋白编码情况与已报道的RBSDV分离物基本一致。 基因组的序列一致性分析和系统进化树表明,RBSDV基因组片段间存在频繁的RNA重排现象;江苏玉米分离物(JS)与湖北玉米分离物(HuB)和河北小麦分离物(HeB)的整体亲缘关系较近,而与安徽分离物(AH-MX8)的整体亲缘关系最远。综合本研究及前人对S8 和S10的研究,RBSDV基因组不同片段的进化中RNA重组的作用不同: S1、S2和S4中没有RNA重组;S3、S5、S6、S8和S10中存在低频率重组;S7 和S9存在较高频率重组,其中S7的高频率重组尤为显著。     

南方水稻黑条矮缩病毒湖南分离物S9片段ORF序列分析

本文克隆了湖南省2011-2013年水稻的15个SRBSDV分离物S9两个基因,并测定了其核苷酸序列,序列分析与比较表明,S9两个ORF序列同源性均大于99.5%。突变位点分析表明,S9编码的两个ORF有3~9个突变位点,但大部分的核酸突变为无义突变。系统发育分析表明,S9编码的两个ORF高度同源,处于相同的的系统发育分支。     

水稻黑条矮缩病控防技术浅析

本文阐述了浙江龙游县水稻黑条矮缩病发生特点,并提出了各种控防技术措施。     

引起玉米粗缩病的水稻黑条矮缩病毒山东分离物的分子特性

 Four isolates of Rice black-streaked dwarf virus (RBSDV) were collected from the maize plants showing rough dwarf symptom in Linyi and Tai'an,Shandong province.The S10 genomic sequences of these isolates were determined and compared with those of 14 other RBSDV isolates.All of the four sequences were 1 801 base pairs (bp) long including the 5'-UTR of 21 bp and the 3'-UTR of 103 bp.They all contained an open reading frame of 1 677 bp (22-1698),encoding the coat protein (CP) of 558 amino acids.The sequences of these four RBSDV isolates and those of the major cp gene of 14 other isolates available in the GenBank were divided into two groups in the phylogenetic tree.Recombination analysis indicated that the isolate Lym2 was likely a recombinant of isolates Lym1 and Zhjs.     

2009年造成湖南省水稻大面积矮缩的是南方水稻黑条矮缩病

2009年在湖南省大面积发生一种水稻病毒病,其症状表现为植株矮缩、叶色深绿、高位分蘖、茎秆出现乳白色或浅褐色点条状突起、茎节上出现气生须根。电镜结果表明:在病株韧皮部可见具斐济病毒特征性晶格状排列球状病毒粒体。病毒基因组S10片段部分序列的相似性比对与系统进化树分析表明,该病害是南方水稻黑条矮缩病。     

湖南省南方水稻黑条矮缩病毒分离物组分S10编码的ORF序列克隆与分析

采用RT-PCR方法扩增并克隆了2013年湖南省水稻上发生的南方水稻黑条矮缩病毒(Southern rice blackstreaked dwarf virus,SRBSDV)S10片段的近全长序列,采用生物信息学软件Mega、RDP和DnaSP分析其编码的开放阅读框(open reading fragment,ORF)遗传多样性。结果表明:2013年湖南省15个SRBSDV分离物S10编码的ORF序列同源性在99.5%以上,不同分离物中存在3~30个突变位点,绝大部分的核酸突变为无义突变,没有发现可能的重组。在系统发育树上,15个分离物聚类为2个分支,其中湖南汉寿的10个分离物和永州的2个分离物(HuNyz12和HuNyz29)与我国浙江分离物聚为1个亚组、湖南永州的其他3个分离物与我国南部的云南、海南等及越南的分离物聚为1个亚组。系统发育结果表明,SRBSDV随传毒介体白背飞虱迁飞从我国南部向北部扩散。     

水稻新病害南方水稻黑条矮缩病发生特点及危害趋势分析

由斐济病毒属(Fijivirus)暂定新种南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)引起的水稻矮缩病,自2001年在广东省阳西县被发现以来,已迅速扩散至我国南方广大稻区。2009年,该病害在我国南部及越南晚季稻上暴发成灾,造成严重的产量损失。本文简介了该病害的症状、危害特点、病原病毒及其传毒介体特征,对其发生趋势进行了讨论,提出了以秧苗期治虫为重点的病害防控应急措施。     

Studies on the epidemiology and yield losses from rice black-streaked dwarf disease in a recent epidemic in Zhejiang province, China

The spread of rice black-streaked dwarf disease, which has emerged as a major problem on winter wheat and the two summer rice crops (early indica and late japonica ) grown in central and southern Zhejiang province, China, is documented from 1995 to 2007. The late japonica crop suffered the most: up to 64 640 ha were affected with estimated losses of c . 120 000 t grain per year. Peak adult numbers of the small brown planthopper vector, Laodelphax striatellus , coincided with the seedling stages of both rice crops and the proportion of the insect population carrying virus increased during 1998–2005. Seedlings with three to four leaves were the most susceptible, whereas plants inoculated after the end of tillering developed few or no symptoms. Disease levels were strongly correlated with numbers of viruliferous vectors. In sowing-date experiments with both rice crops, the earliest sowings had the most disease and suffered the greatest yield losses. With the last sowing date (25 days after the first), there were almost no losses. There were yield losses of 0·80% for every 1% increase in disease incidence in early indica rice and rather more (0·92%) in the late japonica crop. There were large differences in susceptibility between cultivars, indicating the possibility, within currently available germplasm, of using more resistant cultivars to help contain the disease. Changes in cropping practice and in recent winter weather conditions have probably contributed to the emergence of the virus as a major pathogen in eastern China.     

引起玉米粗缩病的水稻黑条矮缩病毒河南分离物的分子特征

<正>目前已报道的造成玉米粗缩病的病原有3种,分别是玉米粗缩病毒(Maize rough dwarf virus,M RDV),水稻黑条矮缩病毒(Rice black-streaked dwarf virus,RBSDV)和南方水稻黑条矮缩病毒(Southern rice black-streaked dwarf virus,SRBSDV)[1]。我国于1954年在新疆和甘肃发现该病,上世纪60年代曾在中东部夏玉米区流行,至70年代以来各玉米产区已陆续发生[2],近年来在黄淮海夏玉米区特别是套播或晚春播、早夏播玉米田发生危害严重。     

影响南方水稻黑条矮缩病发生流行的因子分析

南方水稻黑条矮缩病是近年来在我国南方稻区新发生的一种重要病毒性病害。本文通过大田调查、田间试验并结合气象资料分析,表明品种抗性、种植方式、播种期、播种地段和气象因子等是影响该病发生的主要因素。不同类型水稻的发病率存在显著性差异。杂交稻发病最重,糯稻发病次之,常规稻发病最轻。同一类型的杂交稻不同品种对南方水稻黑条矮缩病的抗(耐)病性存在较大差异。‘特优161’最感病,‘华优638’最抗病。抛栽田比移栽田和直播田发病轻。不同播种期显著影响南方水稻黑条矮缩病的发病率。在化州稻区,播种早的发病重于播种迟的。晚稻秧田靠近早稻本田的田块,南方水稻黑条矮缩病发病重,秧田远离早稻本田的田块发病较轻。6月中旬-7月上旬降水日数、6月下旬-7月上旬相对湿度和8月上旬平均最高气温等气象因子与南方水稻黑条矮缩病发病率关系最密切。     

南方水稻黑条矮缩病毒一步双重RT-PCR 检测技术及其应用

 An one-step dual RT-PCR method was developed for Southern rice black-streaked dwarf virus (SRBSDV) detection from host plants and insect vector white-backed planthopper (Sogatella furcifera Horvath,Hemiptera: Delphacidae), from which two cDNA fragments of the viral genome S5 and S10 were amplified
simultaneously. Two primer pairs, S5-F1 / S5-R2 (5′-ttacaactggagaagcattaacacg-3′ / 5′-atgaggtattgcgtaactgagcc -3′) and S10-oF / S10-oR(5cgcgtcatctcaaactacag -3′ / 5′-tttgtcagcatctaaagcgc -3′), were selectedfrom 40 primer pairs based on SRBSDV genome sequences, and amplified viral S5 ORF1 fragment (819 bp) and cp gene fragment ( 682 bp), respectively. Using total RNA extracts from infected plant leaf tissue or individual planthopper adult as templates, one step RT-PCR reaction in the conditions of annealing temperature of 53℃ with the final concentrations of 240 nmol / L S5-F1 / S5-R2 and 120 nmol / L S10-oF / S10-oR generated a similar yield of two amplicons in argrose electrophoresis analysis. Sequence analysis confirmed the correct
result of the amplification. Additionally, several commercal RNA extraction kits was proven to be fit for the template preparation, and the protocol for total RNA extraction from plant leaf tissue and individual planthopper was simplified by sample grinding direct in eppendorf tube. This method can be used for SRBSDV detection of dried or 70% alcochol soaked planthopper samples stored over one month in room temprature. Key words: Southern rice black-streaked dwarf virus; Sogatella furcifera Horvath