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1.
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   

2.
Tomato leaf curl disease (ToLCD) affected 25% of the tomato crop in Chitrakoot, India and symptomatic leaves were collected for molecular assay. The complete sequences of bipartite begomovirus DNA-A and a betasatellite DNA were amplified. In a sequence analysis, begomovirus DNA-A and betasatellite shared highest sequence identity (91–99%) with Tomato leaf curl New Delhi virus (ToLCNDV) DNA-A and chili leaf curl betasatellite (ChLCB), respectively. The virus was transmitted by whitefly to tomato plants and caused ToLCD symptoms with 70% transmission rate. To our knowledge, this is the first report of the natural occurrence of ToLCNDV and ChLCB in India.  相似文献   

3.
Leaf samples of Cucurbita pepo with yellow mosaic disease symptoms were collected in 2012. Rolling circle amplification and PCR amplification with begomovirus-specific primers confirmed the presence of an Old World bipartite begomovirus, an alphasatellite and a betasatellite. Molecular analysis of full-length sequences showed that Tomato leaf curl New Delhi virus (DNA-A) is associated with its cognate DNA-B, Papaya leaf curl betasatellite and a novel alphasatellite. To the best of our knowledge, this is the first report of an alphasatellite and a betasatellite associated with a bipartite begomovirus.  相似文献   

4.
菜豆金色花叶病毒属(Begomovirus)病毒是热带亚热带地区多种作物的重要病原,杂草作为该属病毒的中间寄主在病害发生流行中具有重要作用。本研究通过克隆、测序和生物信息学分析,对3株具有曲叶症状的凹头苋(Amaranthus lividus)进行菜豆金色花叶病毒属病毒的分离分析。从这些凹头苋中共分离到2种菜豆金色花叶病毒属病毒和3种beta卫星。序列分析显示,其中一种病毒与云南番茄黄化曲叶病毒相似性最高(96%),另一种与中国胜红蓟黄脉病毒相似性最高(96.5%或91%)。Beta卫星的分析显示,其中一种与云南番茄黄化曲叶beta卫星相似性最高(94.3%),另一种与赛葵曲叶beta卫星相似性最高(92%),最后一种与中国番茄曲叶beta卫星相似性最高(91%)。重组分析表明,分离物YN4331-69是一个重组病毒,是由中国胜红蓟黄脉病毒YN4326-60和一个尚未发现的菜豆金色花叶病毒属病毒重组形成。这是首次报道凹头苋被不同的菜豆金色花叶病毒属病毒及其伴随的beta卫星侵染,表明凹头苋是一个适宜该属病毒的中间寄主。  相似文献   

5.
6.
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   

7.
Three begomovirus isolates were obtained from tomato plants showing leaf curl symptoms in Guangxi province of China. Typical begomovirus DNA components representing the three isolates (GX-1, GX-2 and GX-3) were cloned and their full-length sequences were determined to be 2752 nucleotides. Nucleotide identities among the three viral sequences were 98.9–99.7%, but all shared <86.7% nucleotide sequence identity with other reported begomoviruses. The sequence data indicated that GX-1, GX-2 and GX-3 are isolates of a distinct begomovirus species for which the name Tomato leaf curl Guangxi virus (ToLCGXV) is proposed. Further analysis indicated that ToLCGXV probably originated through recombination among viruses related to Ageratum yellow vein virus, Tomato leaf curl China virus and Euphorbia leaf curl virus. PCR and Southern blot analyses demonstrated that isolates GX-1 and GX-2 were associated with DNAβ components, but not isolate GX-3. Sequence comparisons revealed that GX-1 and GX-2 DNAβ components shared the highest sequence identity (86.2%) with that of Tomato yellow leaf curl China virus (TYLCCNV). An infectious construct of ToLCGXV isolate GX-1 (ToLCGXV-GX) was produced and determined to be highly infectious in Nicotiana benthamiana, N. glutinosa, tobacco cvs. Samsun and Xanthi, tomato and Petunia hybrida plants inducing leaf curl and stunting symptoms. Co-inoculation of tomato plants with ToLCGXV-GX and TYLCCNV DNAβ resulted in disease symptoms similar to that caused by ToLCGXV-GX alone or that observed in infected field tomato plants.  相似文献   

8.
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   

9.
为明确南疆温室番茄黄化曲叶病的病毒种类,利用双生病毒的兼并引物通过PCR扩增,对采集的20个番茄病株进行了分子检测.从20个病株中均扩增到约500 bp的目标片段,对其中4株进行克隆和测序,其相互间序列同源性为97.1% ~99.3%,与番茄黄化曲叶病毒(Tomato yellow leaf curl virus,TYLCV)的同源性较高,为98.6% ~ 99.5%.随机选取莎车分离物KS2-5进行全基因组的克隆和测序,KS2-5 DNA全长为2781 nt(序列号:JQ807735),具有典型的双生病毒基因组特征,与TYLCV其它分离物同源性达到98.9%~99.5%,而与其它粉虱传双生病毒的序列同源性较低,为68.3% ~75.5%,表明危害南疆温室番茄的病毒种类为番茄黄化曲叶病毒TYLCV.  相似文献   

10.
ABSTRACT The genome of Tomato leaf curl virus (ToLCV) from Bangalore, India, a whitefly-transmitted geminivirus, was cloned (pIND9) and sequenced. The circular DNA of 2,759 nucleotides (U38239) is organized similarly to that of other begomoviruses with monopartite genomes. Comparison of the nucleotide sequence of pIND9 with other tomato-associated begomoviruses from India (Tomato leaf curl Bangalore virus [ToLCBV, Z48182]) and Tomato leaf curl New Delhi virus-Severe (ToLCNdV-Svr, U15015) showed moderate DNA sequence identities (82 to 87%) between capsid protein (CP) genes but low identities (66 to 67%) for the intergenic regions and the replication-associated protein (Rep) genes (75 to 81% identity). Phylogenetic trees generated with nucleotide sequences of the Rep and CP genes of 26 begomoviruses indicated that this ToLCV is distinct from other begomoviruses and that it may be a recombinant virus derived from at least three different viral lineages. Tomatoes (Lycopersicon esculentum) inoculated with the cloned DNA monomer of ToLCV (pIND9) via particle bombardment developed leaf curling and yellowing symptoms. The virus was transmitted by Bemisia tabaci biotype B from tomatoes infected via particle bombardment to healthy tomatoes and by sap inoculation from infected tomatoes to tomato, Nicotiana benthamiana and N. tabacum. This ToLCV is a distinct member of the genus Begomovirus from India that differs from the previously characterized Tomato leaf curl Sadasivanagar virus isolate Bangalore 1 (L12739), ToLCBV (Z48182), ToLCBV isolate Bangalore 4 (AF165098), and the bipartite ToLCNdV (U15015, U15016). Thus, this ToLCV is named Tomato leaf curl Karnataka virus (ToLCKV).  相似文献   

11.
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   

12.
 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   

13.
Malvastrum leaf curl Guangdong virus is a distinct monopartite begomovirus   总被引:1,自引:0,他引:1  
Virus isolates GD6, GD7, GD8, GD9 and GD10 were obtained from Malvastrum coromandelianum showing leaf curl symptoms in Guangdong Province of China. A specific 500 bp product was consistently detected in total DNA extracts, amplified with universal primers specific for members of the genus Begomovirus. Analysis of their partial DNA sequences revealed that they are isolates of the same begomovirus species, sharing 92·8%–97·1% nucleotide sequence identity. The complete DNA sequences of both GD6 and GD9 were found to be 2767 nucleotides, with all the characteristic features of begomovirus genome organization. The two isolates have less than 85·2% nucleotide sequence identity with other reported begomoviruses. Consequently, GD6 and GD9 are considered to be isolates of a novel begomovirus species, for which the name Malvastrum leaf curl Guangdong virus (MLCuGdV) is proposed. Sequence analyses suggest that MLCuGdV may have arisen by recombination between viruses related to Papaya leaf curl China virus , Tomato leaf curl Philippines virus and other undiscovered virus ancestors. Neither the DNA-B component nor the DNAβ molecule associated with these begomovirus isolates was found. An infectious clone of GD6 was constructed. GD6 efficiently infected Nicotiana benthamiana , N. glutinosa and Petunia hybrida by agro-inoculation, and Malvastrum coromandelianum by whitefly transmission, inducing leaf curling, vein swelling and stunting symptoms. GD6 was also infectious in N. tabacum , but did not induce observable disease symptoms.  相似文献   

14.
Potato (Solanum tuberosum) is one of the important vegetable crops in the world and its production is seriously affected by apical leaf curl disease in northern India. This paper reveals the role of cucurbits in maintaining Tomato leaf curl New Delhi virus (ToLCNDV) and Potato apical leaf curl (PALCD) disease in that region. The affected plants showed severe leaf curling and stunted growth. The begomovirus causing leaf curling and mosaic disease in cucurbits could be easily transmitted by the whitefly to potato crops and develop apical leaf curl disease in northern India. The movement of the virus by whiteflies from cucurbits to potato and tomato is possible because of overlapping of planting and harvesting dates of these crops. The causal virus was identified as a begomovirus on the basis of whitefly transmission, PCR, dot blot hybridization, cloning and sequencing of the coat protein gene. The comparison of full length coat protein gene sequence homology revealed that 90% identity with the coat protein gene of ToLCNDV- [Luffa] isolate and the phylogenetic tree derived from these sequences with other selected begomoviruses formed a close cluster with ToLCNDV isolates. The findings proved that the virus causing disease in cucurbits could easily move to tomato and potato and cause leaf curl disease naturally. This is the first observation on the role of sponge gourd for maintenance of ToLCNDV and serving as a host for PALCD in northern India. The findings indicate that the causal organism is a strain of ToLCNDV.  相似文献   

15.
Whitefly-transmitted begomovirus induces leaf curling and stunting of Mentha arvensis (Kosi) and is responsible for heavy economic losses in and around Lucknow, India. The complete genome was amplified, sequenced and assembled as 2759 bp long, with 1376 bp betasatellite molecule. Because no DNA-B amplicon was detected, the virus was classified as “Old World begomovirus” with the characteristic seven ORFs; two ORFs (AV1, AV2) in the V-sense and five ORFs (AC1–AC5) in the C-sense. BLASTn and phylogenetic analysis showed highest similarity (98%) with chilli leaf curl India virus (ChiLCINV). This is the first report of ChiLCINV infection in M. arvensis (Kosi) from India.  相似文献   

16.
侵染垂花悬铃花的木尔坦棉花曲叶病毒分子特征研究   总被引:2,自引:0,他引:2  
 垂花悬铃花曲叶病是近期在广东发现的一种新病害,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(GD11)DNA-A全长为2 737 nt,具有菜豆金色花叶病毒属病毒基因组典型特征,为闭合环状单链DNA,编码6个ORFs;该序列与木尔坦棉花曲叶病毒(CLCuMV)各分离物序列的相似性均大于89.0%,其中与G6、Okra06及GX1等分离物序列的相似性大于99.0%。该病毒分离物也伴有卫星DNA β分子,其全长为1 348 nt,与CLCuMV各分离物的DNA β序列相似性大于85.0%,其中与G6、Okra06及GX1等DNA β的序列相似性均大于99.0%。因此,侵染广东垂花悬铃花的病毒分离物属于CLCuMV,且与入侵我国的朱槿分离物G6、黄秋葵分离物Okra06及棉花分离物GX1亲缘关系很近。本文首次报道了CLCuMV及其卫星β复合侵染垂花悬铃花。  相似文献   

17.
 2012年5月,江苏南京朱槿上出现一种新的病毒病害,病株表现明显的叶片上卷、叶脉肿大、叶背伴有耳突、植株矮缩等症状。根据其症状及介体发生状况,对其伴随的病毒种类进行研究,结果发现采集的46份典型症状样品体内均可以检测到粉虱传双生病毒,对其基因组DNA-A组分克隆测序后发现其全长2 736 bp,编码6个ORF,BLAST分析显示该病毒与木尔坦棉花曲叶病毒同源性最高(99.9%),是木尔坦棉花曲叶病毒的一个分离物。病样中同时还检测到伴随DNAβ卫星分子,测序结果显示该卫星全长1 346 bp,编码1个ORF,BLAST及聚类分析结果显示该β卫星与木尔坦棉花曲叶病毒β卫星同源性最高(99.7%),为木尔坦棉花曲叶病毒β卫星的一个分离物。这是木尔坦棉花曲叶病毒首次在长江流域的报道。  相似文献   

18.
Whitefly‐transmitted begomoviruses are the most important limiting factor for tomato cultivation in Oman, particularly in the Al‐Batinah region, the major agricultural area of the country. Commercial farms in the Al‐Batinah region were surveyed during January–March 2013. Samples of tomato showing leaf curl disease symptoms typical of begomoviruses were collected and analysed. Full‐length sequences of five clones were shown to have relatively low percentage identity values to known begomoviruses, with the highest (88·6%) to isolates of Tomato leaf curl Oman virus (ToLCOMV), a begomovirus previously reported in Oman, indicating that these represent a newly identified species, for which the name Tomato leaf curl Barka virus (ToLCBrV) is proposed. Four isolates of ToLCBrV were found associated with Tomato leaf curl betasatellite (ToLCB). The five isolates of ToLCBrV characterized in this study were shown to be recombinants, with ToLCOMV as the major parent, and a fragment of Croton yellow vein virus (CrYVV) spanning the 3′ half of the replication‐associated protein. The significance of these findings is discussed.  相似文献   

19.
杂草是菜豆金色花叶病毒属病毒的重要中间寄主,常富集多种该属病毒及其卫星病毒。2014年,在云南红河常见杂草苣荬菜Sonchus arvensis上出现了疑似菜豆金色花叶病毒属病毒病症状。利用克隆、测序和生物信息学分析技术对其所含病毒进行分离鉴定,结果从1株病样中共获得了两条菜豆金色花叶病毒属病毒全序列、两条β卫星全序列和一条α卫星全序列。序列分析显示,两条菜豆金色花叶病毒属病毒全序列与中国胜红蓟黄脉病毒相似性最高,分别为99%和96%,确定为中国胜红蓟黄脉病毒的分离物。两条β卫星全序列与赛葵黄脉β卫星相似性最高,为97%,确定为赛葵黄脉β卫星的一个分离物。α卫星全序列与中国番茄黄化曲叶α卫星相似性最高,为86.3%,是中国番茄黄化曲叶α卫星的一个分离物。这是菜豆金色花叶病毒属病毒病害复合体在中国侵染苣荬菜的首次报道。  相似文献   

20.
Tomato-infecting begomoviruses comprise a complex of monopartite and bipartite virus species that cause severe yield and quality losses worldwide. Therefore, the availability of wide spectrum resistance for begomovirus control is desirable. However, limited sources of resistance are available. In this study, three tomato inbred lines with resistance to bipartite begomoviruses of Brazil were tested for resistance to monopartite begomoviruses associated with the tomato yellow leaf curl disease (TYLCD). Stable resistance to Tomato yellow leaf curl virus was observed either by inoculation with Bemisia tabaci or with Agrobacterium tumefaciens using an infectious clone. The resistance resulted in a complete absence of TYLCD symptoms and restricted virus accumulation. Further studies performed with the line '468-1-1-12' indicated that the resistance was also effective against three other virus species associated with TYLCD, indicating wide spectrum resistance of this source. Quantitative genetics analyses suggested that a major recessive locus with epistatic interactions is controlling the resistance to TYLCD in '468-1-1-12', which could facilitate introgression of this trait into elite tomato lines. The resistance was stable under field conditions with high TYLCD pressure. Mild symptoms could be observed in these conditions, and recovery from disease and from virus infection suggested an active host antiviral defense mechanism. The differential reaction of '468-1-1-12' against a number of TYLCD-associated viruses and artificial chimeras between them allowed to identify a region of the virus genome that presumably contains a virus determinant for breaking the resistance to infection observed in '468-1-1-12'.  相似文献   

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