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1.
2019年山东种植的西葫芦上广泛发生病毒病,症状与之前常发症状有差异,发病植株叶片向下卷曲、黄化,植株矮化。为明确引起此次西葫芦病毒病的病原,我们以田间采集的10份西葫芦病叶为材料,进行PCR扩增并测序。测序结果显示扩增片段核苷酸序列与我国广东的中国南瓜曲叶病毒(SLCCNV)南瓜分离物(MW389917.1)一致性最高。根据同源序列设计引物,经PCR扩增获得SLCCNV全长序列,DNA-A 全长为2 730 bp(OM692270.1)、DNA-B 全长为2 711 bp(OM692269.1),经序列比对发现DNA-A序列与已登录的SLCCNV一致性为89.65%~99.42%,其中与我国广东的SLCCNV-GDHY南瓜分离物(MW389917.1)一致性最高,为99.42%;DNA-B序列与已登录的SLCCNV一致性范围为81.82%~97.29%,其中与我国广东的SLCCNV-GDHY南瓜分离物(MW389918.1)一致性最高,为97.29%。因此推测引发山东西葫芦病毒病的病原物是SLCCNV,由于该病毒是在山东西葫芦上首次发现,将其命名为SLCCNV-SD。前人已报道SLCCNV可侵染南瓜、甜瓜、烟草、番茄等作物,但SLCCNV可侵染西葫芦在国内未见报道。  相似文献   

2.
为明确水茄Solanum torvum植株叶片邹缩、褪绿是否由菜豆金色花叶病毒属病毒侵染引起,从云南省西双版纳傣族自治州田间采集具有疑似感染症状的水茄植株叶片样品,应用菜豆金色花叶病毒属病毒简并引物和特异性引物进行PCR扩增、克隆和测序,通过生物信息软件分析比较其核苷酸序列特征,并对其进行系统发育分析。结果显示,从采集的疑似病叶中共克隆获得了5条菜豆金色花叶病毒属病毒DNA-A全序列和3条DNA-B全序列,经全序列分析发现,侵染水茄的2种菜豆金色花叶病毒属病毒分离物分别属于中国南瓜曲叶病毒(squash leaf curl China virus,SLCCNV)和野茼蒿黄脉病毒(Crassocephalum yellow vein virus,CraYVV)。SLCCNV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒双组分结构特征,与来自泰国的SLCCNV分离物(AB330078)亲缘关系最近,相似性最高达到99.0%;CraYVV水茄分离物的基因组具有典型的菜豆金色花叶病毒属病毒单组分结构特征,与来自云南省景洪市的CraYVV分离物(EF165536)亲缘关系最近,相似性最高达到97.6%。表明水茄是这2种菜豆金色花叶病毒属病毒的新寄主,并首次发现双组分和单组分菜豆金色花叶病毒属病毒可复合侵染水茄。  相似文献   

3.
中国南瓜曲叶病毒(squash leaf curl China virus, SLCCNV)是一种植物单链DNA病毒, 属于双生病毒科Geminiviridae菜豆金色花叶病毒属Begomovirus?在自然条件下该病毒可以侵染多种葫芦科作物, 如南瓜?甜瓜等?该病毒由烟粉虱进行持久性传播, 是影响瓜类产量和品质的重要病害?2021年4月-5月, 安徽省太和县温室大棚中的南瓜叶片出现斑驳?皱缩?卷曲以及植株矮化等现象, 我们采集了具有典型病毒病症状的南瓜病叶样品以及烟粉虱虫体进行RT-PCR鉴定, 结合测序结果鉴定危害南瓜的病毒为SLCCNV?为了进一步明确安徽太和地区SLCCNV的系统进化特征, 我们测定了SLCCNV的外壳蛋白基因(coat protein, CP)序列并进行系统发育分析, 结果表明安徽太和县所检测到的SLCCNV分离物与中国广东和海南地区的分离物亲缘关系较近, 并且与越南?菲律宾?柬埔寨?泰国等一些国家的分离物处于同一个大分支, 存在较小的地域差异性, 而与印度和孟加拉国地区的分离物亲缘关系相对较远, 处于不同的大分支?本研究是安徽省SLCCNV侵染南瓜的首次报道, 期望为该病害的预警和防控提供理论依据?  相似文献   

4.
乔蕊  周雪平  李方方 《植物保护》2024,50(3):266-271
2022年9月在广东省罗定市发现了叶片表现为黄色网状症状的胜红蓟病株。为了明确胜红蓟叶片的黄脉症状是否由双生病毒感染引起, 本研究使用检测双生病毒的简并引物PA/PB进行PCR扩增, 获得约500 bp的片段。根据该序列设计特异性引物扩增并且克隆得到了病毒DNA-A的全基因组序列。通过BLAST比对发现, 获得的DNA-A与中国胜红蓟黄脉病毒(ageratum yellow vein China virus, AYVCNV)海南分离物(OQ421190)的DNA-A的相似性最高, 相似度为98.11%。系统进化树分析显示, 获得的病毒DNA-A与海南分离物(OQ421190)在同一分支, 说明具有较近的亲缘关系。以上研究结果表明侵染胜红蓟的病毒是AYVCNV的分离物。这是关于AYVCNV在广东地区侵染胜红蓟的首次报道, 可为当地病毒病的防控提供参考。  相似文献   

5.
班一云  丁波  周雪平 《植物保护》2017,43(4):134-138
双生病毒是一类在全世界范围内广泛发生的单链环状DNA病毒。本文对从湖南采集到的6例(洋姜、番茄、萝卜、赛葵、甘薯、牵牛花)疑似双生病毒侵染的植物叶片进行了分子鉴定。利用滚环扩增技术(RCA)对样品DNA进行扩增,分别对其RCA产物进行酶切,并将酶切得到的片段测序后进行BLAST比对,结果显示番茄样品中的病毒分离物与番茄黄化曲叶病毒相似性最高(99%),牵牛花样品中的病毒分离物与甘薯卷叶病毒相似性最高(99%),证明这两个分离物是单组分DNA-A双生病毒。这是在湖南省首次发现并报道双生病毒的全核酸序列。  相似文献   

6.
菜豆金色花叶病毒属病毒是一类在全球热带及亚热带地区造成严重经济损失的植物病毒,田间杂草是这类病毒重要的中间寄主。 本研究从云南省玉溪市采集了表现黄脉的青蒿植株,通过PCR扩增、克隆及测序从样品中获得两条菜豆金色花叶病毒属病毒DNA-A全基因组序列,分别为YN6393-23和YN6393-27,其全长均为2 739 bp,相似性为100%。序列分析发现,YN6393-23和YN6393-27的核苷酸序列与烟草曲茎病毒Tobacco curly shoot virus(TbCSV)的分离物YN4584的核苷酸序列相似性最高,为99.45%。根据国际病毒分类委员会对菜豆金色花叶病毒属病毒种的分类标准,全基因组序列相似性大于91%则为同种病毒,表明此病毒分离物为烟草曲茎病毒的一个分离物。这是菜豆金色花叶病毒属病毒侵染青蒿植株的首次报道。  相似文献   

7.
 红麻曲叶病是2012年在海南省海口市发现的一种新病害,病株表现为叶片向上卷曲、叶脉肿大、叶脉变深绿色等症状。PCR检测结果显示,该病样中均存在菜豆金色花叶病毒属病毒。基因克隆及序列分析结果表明,该病毒分离物(HN08)基因组仅含A组分(DNA-A),其全长为2 738 nt,与木尔坦棉花曲叶病毒(CLCuMuV)各分离物的相似性均大于89.0 %,其中与中国各分离物的相似性均大于99.0 %。该病毒分离物也伴随有β卫星分子,其全长为1 346 nt,与CLCuMuV各分离物伴随的β卫星分子(CLCuMuB)序列相似性大于83.0 %,其中与分离物Fz1的序列相似性最高,为99.7 %。构建了HN08 DNA-A及其β卫星分子侵染性克隆,通过农杆菌注射接种红麻,接种后30 d,HN08 DNA-A及其β卫星分子混合接种的红麻植株新出叶片开始产生曲叶症状;接种后60 d,二者混合接种的植株大部分叶片表现为严重的曲叶症状,且与田间自然病株症状相同,而二者各自单独接种的红麻植株没有产生明显的症状。PCR及Southern blot检测进一步证实这些症状是由HN08 DNA-A及其β卫星分子的共同侵染引起的。因此,海南红麻曲叶病是由CLCuMuV及其伴随的β卫星分子(CLCuMuB)共同侵染引起的。本文首次报道了红麻是CLCuMuV自然新寄主。  相似文献   

8.
为明确假酸浆Nicandra physalodes叶片黄化、皱缩症状是否由菜豆金色花叶病毒属病毒侵染引起,本研究利用分子检测方法和生物信息学技术鉴定了假酸浆样品中的病毒种类。从采集的病样中克隆并获得了2条菜豆金色花叶病毒属病毒DNA-A全序列和1条beta卫星全序列,经全序列分析发现,该双生病毒的两条DNA-A全序列与泰国番茄黄化曲叶病毒(tomato yellow leaf curl Thailand virus, TYLCTHV)云南分离物TYLCTHV-YN1732一致性最高,达99.3%,亲缘关系较近;beta卫星的全序列与云南番茄曲叶beta卫星(tomato leaf curl Yunnan betasatellite, TLCYnB)的分离物YN5230一致性最高,达99.3%,亲缘关系较近。重组分析显示,假酸浆上分离的TYLCTHV-YN5735-12是一个重组病毒,有两个重组事件,一个主要发生在AV1的编码区,由中国番茄黄化曲叶病毒(tomato yellow leaf curl China virus, TYLCCNV)和广西大戟曲叶病毒(euphorbia lea...  相似文献   

9.
为明确引起我国山西晋中地区苘麻叶片表现皱缩和花叶症状的病原物及其基因组分子特征, 本研究利用双生病毒简并引物扩增获得病毒基因组部分序列,经测序、比对后设计特异性引物扩增病毒基因组序列, 进而通过生物信息学方法构建系统发育树并进行序列分析。结果表明:引起苘麻叶片皱缩、花叶的病原物为番茄黄化曲叶病毒(tomato yellow leaf curl virus, TYLCV), 将该分离物命名为TYLCV-Abu, GenBank登录号为OP293347, 但未扩增到β卫星。该病毒DNA-A基因组全长为2 782 bp, 含有6个开放阅读框。TYLCV-Abu分离物与TYLCV茄子分离物KSQ1-3(GenBank登录号KC428753)的核苷酸序列一致性最高, 为98.99%, 其中C4和V2编码的蛋白变异较大。重组结果分析显示,分离物TYLCV-Abu是由TYLCV-F(GenBank登录号KY971326)和TYLCV-KSQ1-3重组得到, 重组区域为其基因组2 617-2 782 nt区域。这是首次从苘麻样品中扩增到TYLCV全基因组序列并进行分析。  相似文献   

10.
2012年秋季, 在山东泰安番茄主要种植区中采集到叶片褪绿, 叶脉颜色变深的疑似番茄褪绿病毒病和番茄侵染性褪绿病毒病的番茄样品。利用番茄褪绿病毒(Tomato chlorosis virus, ToCV)的特异引物 ToCV1/ToCV2和番茄侵染性褪绿病毒(Tomato infectious chlorosis virus, TICV)的特异引物TICV1/TICV2分别对样品进行扩增, 最后仅得到利用引物 ToCV1/ToCV2 扩增的101 bp 的核苷酸序列, 对该核苷酸序列克隆并测序。序列比对表明, 山东泰安地区分离物与已登录的番茄褪绿病毒(ToCV)分离物相似性都在99%以上。随后, 对山东泰安种植区ToCV番茄分离物进行外壳蛋白(CP)及热激蛋白(HSP70)序列的扩增、克隆和测序(GenBank登录号KC812620/KC812625), 经NCBI BLAST比对发现, 目的序列与番茄褪绿病毒日本番茄分离物ToCV Japan/Tochigi (GenBank登录号AB513442/AB513443)相似性最高为99%, 同属于毛型病毒属的番茄褪绿病毒, 这是首次明确山东地区番茄受到番茄褪绿病毒的侵染。  相似文献   

11.
Euphorbia mosaic virus (EuMV), a tentative species within the genus Begomovirus, was isolated from Euphorbia heterophylla plants growing in the Yucatan Peninsula, Mexico. The complete bipartite genome was cloned from total DNA extracts and the nucleotide (nt) sequence was determined. The DNA-A sequence of the EuMV-Yucatan Peninsula (EuMV-YP) isolate shared 95% nt identity with the partially characterized type EuMV isolate from Puerto Rico. The EuMV-YP genome organization was like that of other New World, bipartite begomoviruses. The DNA-A component was 2613 nt in size, while the DNA-B component was 2602 nt long. The 165-nt common region (CR) sequence for the DNA-A and DNA-B components shared a lower than expected nt identity of 86%. The organization and iterons of the putative AC1 binding site of EuMV-YP were similar to those of begomoviruses in the Squash leaf curl virus (SLCV) clade. Characteristic disease symptoms were reproduced in E. heterophylla plants inoculated at the seedling stage using the cloned viral DNA-A and DNA-B components, confirming disease aetiology. Results of an experimental host-range study for EuMV-YP indicated that it infected at least five species in three plant families, including the Euphorbiaceae ( E. heterophylla ), Solanaceae ( Datura stramonium , pepper, tomato) and Fabaceae (bean). Phylogenetic analysis of the DNA-A and DNA-B components indicated that EuMV-YP is a New World begomovirus and that it is a new member of the SLCV clade.  相似文献   

12.
ABSTRACT The biological and molecular properties of Tomato leaf curl Gujarat virus from Varanasi, India (ToLCGV-[Var]) were characterized. ToLCGV-[Var] could be transmitted by grafting and through whitefly transmission in a persistent manner. The full-length genome of DNA-A and DNA-B of ToLCGV-[Var] was cloned in pUC18. Sequence analysis revealed that DNA-A (AY190290) is 2,757 bp and DNA-B (AY190291) is 2,688 bp in length. ToLCGV-[Var] could infect and cause symptoms in tomato, pepper, Nicotiana benthamiana, and N. tabacum when partial tandem dimeric constructs of DNA-A and DNA-B were co-inoculated by particle bombardment. DNA-A alone also is infectious, but symptoms were milder and took longer to develop. ToLCGV-Var virus can be transmitted through sap inoculation from infected tomato plants to the above-mentioned hosts causing the same symptoms. Open reading frames (ORFs) in both DNA-A and DNA-B are organized similarly to those in other begomoviruses. DNA-A and DNA-B share a common region of 155 bp with only 60% sequence identity. DNA-B of ToLCGV-[Var] shares overall 80% identity with DNA-B of Tomato leaf curl New Delhi virus-Severe (ToLCNDV-Svr) and 75% with ToLCNDV-[Lucknow] (ToLCNDV-[Luc]). Comparison of DNA-A sequence with different begomoviruses indicates that ToLCGV-[Var] shares 84% identity with Tomato leaf curl Karnataka virus (ToLCKV) and 66% with ToLCNDV-Svr. ToLCGV-[Var] shares a maximum of 98% identity with another isolate of the same region (ToLCGV-[Mir]; AF449999) and 97% identity with one isolate from Gujarat (ToLCGV-[Vad]; AF413671). All three viruses belong to the same species that is distinct from all the other geminivirus species described so far in the genus Begomovirus of the family Geminiviridae. The name Tomato leaf curl Gujarat virus is proposed because the first sequence was taken from an isolate of Gujarat, India.  相似文献   

13.
ABSTRACT Bean calico mosaic virus (BCMoV), a whitefly-transmitted geminivirus from Sonora, Mexico, was purified, and the genome components were cloned and sequenced. Purified viral fractions and cloned genome components were infectious by biolistic inoculation to bean, completing Koch's postulates for both. The B biotype of the whitefly Bemisia tabaci efficiently transmitted both native virus and progeny virus derived from cloned DNA inoculum. Host ranges of native virus and of progeny virus derived from cloned DNA were identical based upon whitefly and biolistic mediated transmission, respectively. BCMoV has a relatively wide experimental host range among begomoviruses known to infect bean, encompassing genera and species within the Fabaceae, Malvaceae, and Solanaceae. BCMoV has a bipartite genome, as do other New World begomoviruses. BCMoV DNA-A shared highest nucleotide sequence identities with squash leaf curl virus-E strain (SLCV-E) and cabbage leaf curl virus (CaLCV) at 80.1 and 80.7%, respectively. BCMoV DNA-B shared highest nucleotide sequence identity with SLCV-E at 70.7%. The common region (CR) sequences of BCMoV and SLCV-E are 73 to 76% identical; however, modular cis-acting elements within the CR involved in replication origin function and recognition are 100% conserved. Phy-logenetic analysis indicated that BCMoV DNA-A shares a most recent common ancestor with the DNA-A of two viruses that also occur in the Sonoran Desert, SLCV-E and Texas pepper virus (TPV-TAM), and CaLCV from Florida. In contrast, a phylogenetic analysis indicated that BCMoV DNA-B shares a most recent common ancestor with SLCV-E; whereas DNA-B of CaLCV clustered in a separate clade with pepper hausteco virus. Collectively, biological and molecular characteristics indicate that BCMoV is a distinct begomovirus species with the northernmost distribution of any begomovirus isolated from bean in the Americas. Furthermore, the phylogenetic relationships of begomovirus cognate components are not necessarily identical, suggesting that DNA-A and DNA-B of some begomoviruses may have different evolutionary histories.  相似文献   

14.
 从采集于海南儋州地区表现黄脉症状的长蒴母草(Lindernia anagallis)上分离到病毒分离物L2, DNA-A全序列分析结果表明, 全长2739个核苷酸(nt)(GenBank登录号:AY795900), 共编码6个ORF, 其中病毒链编码AV1(CP)、AV2, 互补链编码AC1、AC2、AC3、AC4。利用BLAST程序对DNA-A进行分析表明, 与L2 DNA-A有同源关系的病毒均为双生病毒科(Geminiviridae)菜豆金色黄花叶病毒属(Begomovirus)成员。进一步比较发现, L2 DNA-A与我国广东报道的广东番茄曲叶病毒(Tomato leaf curl Guangdong virus, ToLCGuV)(AY602165)全基因组核苷酸序列的同源性最近, 仅为77.0%, 说明L2为Begomovirus中的一个新种, 命名为长蒴母草黄脉病毒(Lindernia anagallis yellow vein virus, LAYVV)。与L2的IR区及各基因编码的氨基酸序列有最高同源性的病毒均来源于亚洲。利用DNA-B特异引物和DNA-β的特异引物, 均未检测到DNA-B和卫星DNA-β的存在。  相似文献   

15.
Genomic characterization using nonradioactive probes, polymerase chain reaction with degenerate primers for whitefly transmitted geminiviruses and nucleotide sequencing were used to describe a new bipartite geminivirus, associated with dwarfing and leaf curling of tomatoes and peppers in Jamaica. Partial DNA-A and DNA-B clones were obtained. DNA sequence analysis showed that tomato and pepper samples have a similar geminivirus associated with them. Nucleotide sequence identity > 92% between the common regions of DNA-A and DNA-B confirmed the bipartite nature of the Jamaican geminivirus isolates. Nucleotide sequence comparisons of DNA-A and DNA-B with those of geminiviruses representing the major phylogenetic groups of Western Hemisphere geminiviruses showed the greatest similarity to potato yellow mosaic virus and members of the Abutilon mosaic virus cluster of geminiviruses. This new virus is given the name tomato dwarf leaf curl virus (TDLCV) because of the dwarfing and leaf curling symptoms associated with infected tomato plants. Polymerase chain reaction and Southern hybridization showed mixed infections of TDLCV with tomato yellow leaf curl virus from Israel in 16% of the field samples of tomatoes and peppers.  相似文献   

16.
苏秀  陈莎  周湘  龙奎  刘颖  郭恺 《植物病理学报》2019,49(3):424-427
<正>大青(Clerodendrum cyrtophyllum Turcz),别名路边青、淡婆婆、臭叶树,为马鞭草科(Verbena officinalis Linn)大青属(Clerodendrum)野生落叶灌木,常生长在山地林下、平原、丘陵和溪谷旁,我国江苏、安徽、河北、河南、浙江等地为主产区。大青用途广泛,叶片可萃取靛蓝作为染料,嫩叶可食用,  相似文献   

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