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2.
草莓轻型黄边病毒3'末端序列多态性研究 总被引：1，自引：0，他引：1
The 930 bp segment in 3' terminal region of Strawberry mild yellow edge virus(SMYEV) genome was amplified by 3' rapid amplification of cDNA ends(RACE).Ten Chinese isolates were sequenced,and 7 of them were the same.Nucleotide and amino acid identities and phylogenesis were analyzed between Chinese isolates and 24 isolates from other regions of the world.Sequence analysis of the 878 nt stretch within 3' terminal region of SMYEV genome showed that nucleotide acid identities ranged from 79.5% to 100%,deduced amino acid sequences of coat protein gene identity were 86.4% to 100%.Phylogenetic analysis showed that all isolates of SMYEV fell into four clades.To a certain extent,the clades were related with the geological distribution of SMYEV.Chinese isolates SY01 and SY04 lay in the same clade with European and American isolates,but formed a small separate branch.Isolates SY03 and SY02,derived from Fragaria×ananassa cv.Changhong-2 and F.pentaphylla respectively,had a far relationship with other isolates and fell into one clade.They were likely to be the special isolates that existed only in China. 相似文献
3.
The tripe gene block (TGB)genes of Barley stripe mosaic virus China strain (BSMV-CH)were amplified from cDNA of BSMV-CH RNAβ (GenBank accession No:AY789694)by PCR with special primer pairs, and cloned into pMD18-T vector for sequencing. Analysis of the sequences showed that the full length of BSMV-CH TGB1, TGB2 and TGB3 were 1 539, 396 and 468 bp, with deduced 512, 131 and 155 amino acids, respectively. BSMV-CH TGB1 shared 94.1%-95.4% nucleotide identities and 91.0%-94.5% amino acid identities with that of other BSMV strains, BSMV-CH TGB2 shared 96.5%-97.2% nucleotide identities and 98.5%-99.2% amino acid identities with that of other BSMV strains, and BSMV-CH TGB3 shared 95.7%-96.6% nucleotide identities and 94.2%-96.8% amino acid identities with that of other BSMV strains. Phylogenetic tree based on the amino acid of Hordeiviruses TGB genes showed that BSMV-CH was relatively closed to CV17 in genetic relationship. Therefore, BSMV-CH was deduced to be a recombined strain of CV17 and CV42. 相似文献
4.
The gene of RNA-dependent RNA polymerase (RdRP) was cloned by RT-PCR from Sugarcane yellow leaf virus-Fuzhou isolate (CHN-FJ1) and then cloned into pMD18-T vector. The sequence showed that the fragment comprised 1 212 nucleotides including part gene of ORF1 and ORF2. The ORF2 was involved the RdRP gene consisted of 995 nucleotides and encoded putative protein of 331 amino acids. Compared the nucleo-tide sequence and encoded putative protein of CHN-FJ1 with the other isolates from different countries, they shared the homology above 92.0%. Phylogenetic tree suggested that the sixteen isolates were classified into four types according to the amino acid sequence of the RdRP. One of the groups contained CHN-FJ1 and the other isolates from China, American, Brazil, Australia and Colombia;however, there was the closest relation between CHN-FJ1 and BRA-YL1 isolate from Brazil. 相似文献
5.
甘蔗花叶病毒福建分离物外壳蛋白基因的克隆及序列分析 总被引：3，自引：0，他引：3
A fujian isolate of Sugarcane mosaic virus named SCMV-FJ was isolated from infected sugarcane. Cloning and sequence analysis of the coat protein gene of this isolate was carried out. A pair of primers was designed and synthesized based on the nucleotide sequences of coat protein genes of sugarcane mosaic viruses reported. The coat protein gene of SCMV-FJ was amplified from the extracted total RNA of the infected sugarcane by using RT-PCR, and cloned into the pMD18-T vector. The sequencing result indicated that the cloned segment included a 1137 bp open reading frame(ORF) and a 228 bp 3' untranslated region, in which the ORF comprised the whole coat protein and part of the nuclear inclusion b. The nucleotide and the deduced amino acid sequences of the coat protein gene were compared with those of the other isolates or strains of SCMV subgroup reported in GenBank. The result showed that it shares 56.8%-97.1% and 55.3%-99.4% homology in nucleotide and the putative amino acid sequences, respectively, with the highest amino acid homology of 99.4% with SCMV-D. Thus it was identified as a SCMV-D. This experiment provided a rapid, sensitive and relatively inexpensive method for RT-PCR detection of SCMV. At the same time, the cloning of SCMV-FJ coat protein gene provided the foundation for plant gene engineering against SCMV. 相似文献
6.
小西葫芦黄花叶病毒山东南瓜分离物的分子特性 总被引：2，自引：0，他引：2
Zucchini yellow mosaic virus (ZYMV) was detected by RT-PCR from pumpkin (Cucurbita moschata) plant showing yellowing and mosaic symptom from Liaocheng, Shandong Province. The 3'-termial 1 684 bp genomic sequence covered 633 bp of NIb encoding sequence, 840 bp of cp gene and 211 bp of 3'-untranslated region of the isolate ZYMV-Liaocheng was determined. The cp gene of ZYMV-Liaocheng shared identities of 81.4%-98.8% and 89.4%-99.5% at nucleotide and amino acid levels, respectively, with other ZYMV sequences available in the GenBank. Phylogenetic analysis indicated that ZYMV could be clustered to 6 genotypes. ZYMV-Liaocheng belonged to genotypeⅠ, which contained isolates from Asia, Europe and America. Genotypes Ⅲ and Ⅴ were unique and contained only isolates from East Asia. The isolates from East Asia had the highest variability. 相似文献
7.
茄科蔬菜立枯丝核菌的融合群鉴定 总被引：3，自引：0，他引：3
Sixty-five samples were collected from rhizosphere soil, hot pepper and tomato plants showing damping-off, root rot and stem rot in Taian, Shouguang of Shandong province and Zhouzhi, Taibai of Shaanxi province. Thirty-nine Rhizoctonia solani isolates were obtained from these samples. The results of anastomosis group (AG) identification and sequence analysis of 5.8S rDNA-ITS of the isolates showed that thirty-six isolates (92.3%) belonged to AG-4, while only three (7.7%) belonged to AG-5. The isolates of AG4 could further be divided into two subgroups of AG4-HG-Ⅰ and AG4-HG-Ⅲ. The 5.8S rDNA-ITS sequences of the selected isolates of the two subgroups had the 99%-100% identity with standard isolates of AG4-HG-Ⅰ and AG4-HG-Ⅲ (from GenBank). Among the analyzed isolates, AG4-HG-Ⅰ subgroup was the dominant with the frequency of 79.5%. Subgroup AG-4-HG-Ⅲ with the frequency of 12.8% was the second. This is the first report that subgroup AG4-HG-Ⅲ of R. solani isolated from Solanaceae vegetable crops in China. 相似文献
9.
进境玉米种子携带玉米褪绿斑驳病毒的检测与鉴定 总被引：3，自引：0，他引：3
Imported maize seeds were planted in a quarantine greenhouse and two seedlings with chlorotic mottle symptoms were observed. The ELISA results showed that the two seedlings reacted positively with antibody against Maize chlorotic mottle virus (MCMV). The positive samples were further identified by RT-PCR method and the 711 bp size target bands were amplified specifically. The similarity range of nucleotide sequence of both RT-PCR product and six MCMV coat protein genes was 97%-99%. The amino acid sequence homology was 97.88%-99.89%. Based on the above results, the virus was identified as MCMV. It was the first time that MCMV was intercepted from imported maize seeds. 相似文献
10.
YN80 was isolated from Amorphophallus rivieri Durieu showing mosaic and crinkle symptoms in Songming, Yunnan province. Flexuous filamentous particles were found in diseased leave sap and pinwheel inclusion bodies were found in the leave tissue. YN80 had positive reaction to universal antibody of Potyvirus by DAS-ELISA. 3'-terminal sequence of YN80 was cloned and sequenced. cp gene of YN80 consisted of 987 nt, encoded 328 aa (36.1 kDa). Sequence analysis showed that YN80 shared the highest identity (97.0%)with CP amino acid sequence of Dasheen mosaic virus (DsMV). These data indicated that YN80 was an isolate of DsMV. This is the first molecular identification of A. rivieri Durieu isolate of DsMV in China. 相似文献
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12.
An isolate of PVS, HZ00P1, was obtained from nature infected Solanum tuberosum from Hang-zhou, Zhejiang province. It was primarily identified by DAS-ELISA and host reaction tests. 3' end partial sequence of the virus isolate was cloned. Nucleic acid sequence of the cp gene and deduced cp amino acid se-quence alignments were compared with 16 other isolates registered in GenBank. The results showed that the 17 PVS isolates were divided into two groups. Among them, two Andean isolates were classified to group Ⅱ, HZ00P1 and the other 14 isolates were assigned to group Ⅰ. Compared with other PVS isolates in group Ⅰ, PVS HZ00P1 had 93.1%-98.1% and 95.9%-99.3% homologies of nucleotide sequence and deduced amino acid sequence respectively, whereas its similarities to the group Ⅱwere 81.4%-81.7% and 93.5%-93.9%. A survey of PVS occurrence in Zhejiang province was achieved by RNA spot hybridization (RSH). Among the 30 samples collected from different areas of the province, 26.7% were found to have the infection of PVS. It is concluded that PVS has become a common virus in Zhejiang province. 相似文献
13.
核苷酸序列分析结果表明,小麦黄色花叶病毒(W YMV)不同分离物的外壳蛋白基因存在一定的差异。邓州分离物CP基因在其31~33nt处均缺失了3个核苷酸,其余分离物与潢川分离物及日本分离物长度一致,均为882nt。不同分离物CP基因核苷酸序列同源性为97.3%~98.9%,由此推导的氨基酸序列同源性为97.6%~99.3%,外壳蛋白N末端的110个氨基酸和C末端的55个氨基酸在各个分离物间是高度保守的。潢川分离物有5个氨基酸与其它5个分离物明显不同。WYMV不同分离物外壳蛋白序列分析结果进一步确认了WYMV与WSSMV为Bymovirus属的2种不同病毒。 相似文献
14.
对山东省侵染马铃薯的一个马铃薯X病毒(PVX)分离物PVX—SD1的外壳蛋白(CP)基因进行了克隆和序列分析。以提纯的病毒RNA为模板，应用RT-PCR扩增目的基因，通过常规的基因克隆法将扩增的CP基因导入pUC19载体，测序。结果表明，PVX—SD1的CP基因长719bp，可编码248个氨基酸；与Gen—Bank中报道的15个有代表性的株系或分离物相比较，核苷酸同源性在80．1％-99．7％，氨基酸同源性在89．8％-100％；与欧洲株系UK3仅1个核苷酸不同，同源性为99．7％，氨基酸同源性达100％，表明它们可能为同一株系，属于X^3组． 相似文献
15.
葡萄卷叶伴随病毒2号和3号辽宁分离物部分基因组的序列分析 总被引：3，自引：1，他引：2
将采自辽宁兴城地区在生长期间具典型卷叶病症状的金星无核(Venus Seedless)葡萄品种休眠枝条,用RT-PCR检测4种葡萄卷叶伴随病毒(Grapevine leafroll-associated viruses,GLRaVs),扩增得到了葡萄卷叶伴随病毒2号(GLRaV-2)和葡萄卷叶伴随病毒3号(GLRaV-3)两种病毒的主要外壳蛋白(major coat protein,CP)基因的完整序列(GenBank登录号分别为FJ786017和FJ786016)。这表明该葡萄植株受到了GLRaV-2和GLRaV-3辽宁分离物(GLRaV-2-LN和GLRaV-3-LN)的复合侵染。根据检测结果,克隆了GLRaV-2-LN基因组3'端CPm (minor capsid protein)、p19(19-kDa protein)和p24(24-kDa protein)基因(GenBank登录号分别为FJ786018、FJ786019和FJ786018)。序列分析表明,GLRaV-3-LN的CP基因全长942 nt,与已报道的国内外其它分离物CP基因全序列相比,核苷酸序列同源性为89.8%~91.8%,由此推导的氨基酸序列同源性为94.9%~97.4%。GLRaV-2-LN的CP、CPm、p19和p24基因全长分别为597 nt、672 nt、486 nt和618 nt。与国外报道的几个分离物的相应蛋白基因全序列相比,核苷酸序列同源性分别为88.3%~100.0%、78.7%~99.9%、75.1%~99.4%和87.5%~99.5%;由此推导的氨基酸序列同源性分别为92.9%~100.0%、89.2%~100.0%、73.9%~99.4%和89.3%~99.0%。 相似文献
16.
通过高通量测序和RT-PCR扩增,克隆并分析蟹爪兰X病毒(Zygocactus virus X,ZyVX)贵州分离物ZyVX-GZ基因组序列。对表现病毒病症状的红肉火龙果植株进行高通量测序,获得4条ZyVX相关contig。RT-PCR扩增并拼接获得ZyVX-GZ基因组,全长为 6 567 nt,5′-UTR和 3′-UTR分别为60 nt和70 nt。含有5个ORF,分别编码复制酶蛋白、TGB1、TGB2、TGB3蛋白和外壳蛋白。ZyVX-GZ与P39和B1分离物基因组序列一致性均为91%。TGB1-3、外壳蛋白基因与大多数分离物核苷酸和推导的氨基酸序列一致性分别为95%～99%和96%～100%;复制酶基因的核苷酸和推导的氨基酸序列一致性分别为80%～89%和92%～97%。系统进化分析表明,ZyVX群体的遗传分化与寄主种类、地理分布不存在相关性。本研究结果丰富了ZyVX群体的基因组序列信息,为ZyVX的监测和防控提供了基础。 相似文献
17.
西瓜花叶病毒2号新疆昌吉分离物外壳蛋白基因核苷酸序列分析 总被引：6，自引：1，他引：5
利用RT-PCR从新疆昌吉地区表现花叶、疱斑、扭曲等症状的南瓜病株上检测到西瓜花叶病毒2号新疆昌吉分离物(简称WMV-2-XJ-CJ),并测定了该分离物外壳蛋白(CP)基因序列。序列分析表明,新疆昌吉分离物CP基因全长850个核苷酸,编码197个氨基酸。与国内外报道的12个WMV-2CP基因相比,其核苷酸序列同源性为92.6%~98.3%,由此推导的氨基酸序列同源性为94.7%~99.3%。新疆昌吉分离物在CP N'端可变区明显不同于国内外报道的核苷酸序列。WMV-2新疆昌吉分离物与日本和郑州分离物较其它国家和地区的分离物多出6个核苷酸,但其核苷酸及其推导的氨基酸序列差异较大。新疆昌吉分离物外壳蛋白有2个氨基酸残基明显不同于其它分离物,其中蚜传株系的特征结构域DAG突变为DAE。 相似文献
18.
为明确黄瓜花叶病毒（cucumber mosaic virus,CMV）甜瓜分离物的分子变异情况及其侵染性,对2个甜瓜分离物CH99和XH18的基因组进行克隆、测序和分析,并通过构建全长cDNA克隆分析其侵染性。结果显示,黄瓜花叶病毒甜瓜CH99分离物3条RNA长度分别为3 356、3 049和2 211 nt,甜瓜XH18分离物3条RNA长度分别为3 381、3 048和2 217 nt。分离物CH99与XH18的核苷酸序列一致性为89.40%～95.80%,氨基酸序列一致性为90.00%～97.80%,CH99分离物与其他CMV分离物的核苷酸和氨基酸序列一致性平均值分别为79.23%～89.29%和73.52%～93.90%,XH18分离物与其他CMV分离物的核苷酸和氨基酸序列一致性平均值分别为79.81%～89.83%和74.02%～95.14%。遗传发育分析显示,这2个分离物均属于亚组IB成员。接种试验结果显示,分离物CH99和XH18的侵染性克隆构建成功,这2个分离物均能系统侵染本生烟、甜瓜和黄瓜,并在本生烟和甜瓜上引起较严重的症状,在黄瓜上引起的症状较弱,而二者均不能侵染西... 相似文献
19.
中国北方四省小麦丛矮病病原鉴定 总被引：1，自引：0，他引：1
小麦丛矮病是由灰飞虱(Laodelphax striatellus Fallén)传播的一种病毒病,具有流行性和暴发性,可致小麦减产甚至绝收.小麦丛矮病的典型症状表现为叶片有黄绿相间条纹,分蘖增多,植株矮化,形成明显的丛矮状.冬前感病株大部分不能越冬而死亡,冬前未显症和早春及拔节后感病植株上部叶片显条纹,一般不能抽穗而提早枯死,有的能抽穗,但穗小籽粒秕瘦,对产量影响较大.研究结果表明小麦丛矮病的病原为北方禾谷花叶病毒(Northern cereal mosaic virus,NCMV)[1-3].小麦丛矮病在我国分布较广[4,5].为了进一步明确小麦丛矮病病原发生、分布及其变异,2010年4月在河北、河南、山东和山西省多地采集标样,采用RTPCR方法对各地标样进行了鉴定与分析,以期为病害防治提供理论依据. 相似文献
20.
甘薯脉花叶病毒外壳蛋白基因在大肠杆菌中的表达及其抗血清的制备 总被引：1，自引：1，他引：0
根据已报道的甘薯脉花叶病毒(Sweet potato vein mosaic virus,SPVMV)外壳蛋白（CP）基因的核苷酸序列合成引物,利用RT-PCR方法克隆了SPVMV河南分离物（SPVMV-HN）基因组3′端1.8 kb的基因片段,包括部分NIb 基因序列和完整的CP基因及3′端非编码区序列(3′UTR)。序列分析表明,SPVMV-HN的CP基因由996个核苷酸组成（GenBank登录号为FJ687211）,编码332个氨基酸残基。与已发表的SPVMV其他分离物相比,其推导的氨基酸序列一致性为95.2%～98.5%,与 SPVMV广东分离物的氨基酸序列一致性为97.9%。将CP基因克隆到原核表达载体pET-28a（+）上,SDS-PAGE分析表明,经IPTG诱导,CP基因在大肠杆菌BL21(DE3) pLysS中得到了高效表达。以表达的蛋白为抗原,免疫家兔,制备了SPVMV外壳蛋白的特异性抗血清。ACP-ELISA检测结果表明,制备的抗血清可用于田间甘薯样品的检测。利用SPVMV的抗血清,对采自全国14个省（市）的田间甘薯样品以及嫁接的巴西牵牛样品进行了检测,结果表明,SPVMV在我国甘薯上普遍存在。 相似文献