植物病理学报
植物病理學報
식물병이학보
ACTA PHYTOPATHOLOGICA SINICA
2009年
5期
458-465
,共8页
王萌%费菲%周涛%程玉琴%范在丰
王萌%費菲%週濤%程玉琴%範在豐
왕맹%비비%주도%정옥금%범재봉
葡萄卷叶伴随病毒%RT-PCR%克隆%序列分析
葡萄捲葉伴隨病毒%RT-PCR%剋隆%序列分析
포도권협반수병독%RT-PCR%극륭%서렬분석
Grapevine leafroll-associated virus%RT-PCR%cloning%sequencing
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,与已报道的同内外其它分离物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%.
將採自遼寧興城地區在生長期間具典型捲葉病癥狀的金星無覈(Venus Seedless)葡萄品種休眠枝條,用RT-PCR檢測4種葡萄捲葉伴隨病毒(Grapevine leafroll-associated viruses,GLRaVs),擴增得到瞭葡萄捲葉伴隨病毒2號(GLRaV-2)和葡萄捲葉伴隨病毒3號(GLRaV-3)兩種病毒的主要外殼蛋白(major coat protein,CP)基因的完整序列(GenBank登錄號分彆為FJ786017和FJ786016).這錶明該葡萄植株受到瞭GLRaV-2和GLRaV-3遼寧分離物(GLRaV-2-LN和GLRaV-3-LN)的複閤侵染.根據檢測結果,剋隆瞭GLRaV-2-LN基因組3'耑CPm(minor capsid protein)、p19(19-kDa protein)和p24(24-kDa protein)基因(GenBank登錄號分彆為FJ786018、FJ786019和FJ786018).序列分析錶明,GLRaV-3-LN的CP基因全長942 nt,與已報道的同內外其它分離物CP基因全序列相比,覈苷痠序列同源性為89.8%~91.8%,由此推導的氨基痠序列同源性為94.9%~97.4%.GLRaV-2-LN的CP、CPm、p19和p24基因全長分彆為597 nt、672 nt、486 nt和618 nt.與國外報道的幾箇分離物的相應蛋白基因全序列相比,覈苷痠序列同源性分彆為88.3%~100.0%、78.7%~99.9%、75.1%~99.4%和87.5%~99.5%;由此推導的氨基痠序列同源性分彆為92.9%~100.0%、89.2%~100.0%、73.9%~99.4%和89.3%~99.0%.
장채자료녕흥성지구재생장기간구전형권협병증상적금성무핵(Venus Seedless)포도품충휴면지조,용RT-PCR검측4충포도권협반수병독(Grapevine leafroll-associated viruses,GLRaVs),확증득도료포도권협반수병독2호(GLRaV-2)화포도권협반수병독3호(GLRaV-3)량충병독적주요외각단백(major coat protein,CP)기인적완정서렬(GenBank등록호분별위FJ786017화FJ786016).저표명해포도식주수도료GLRaV-2화GLRaV-3료녕분리물(GLRaV-2-LN화GLRaV-3-LN)적복합침염.근거검측결과,극륭료GLRaV-2-LN기인조3'단CPm(minor capsid protein)、p19(19-kDa protein)화p24(24-kDa protein)기인(GenBank등록호분별위FJ786018、FJ786019화FJ786018).서렬분석표명,GLRaV-3-LN적CP기인전장942 nt,여이보도적동내외기타분리물CP기인전서렬상비,핵감산서렬동원성위89.8%~91.8%,유차추도적안기산서렬동원성위94.9%~97.4%.GLRaV-2-LN적CP、CPm、p19화p24기인전장분별위597 nt、672 nt、486 nt화618 nt.여국외보도적궤개분리물적상응단백기인전서렬상비,핵감산서렬동원성분별위88.3%~100.0%、78.7%~99.9%、75.1%~99.4%화87.5%~99.5%;유차추도적안기산서렬동원성분별위92.9%~100.0%、89.2%~100.0%、73.9%~99.4%화89.3%~99.0%.
The dormant cutting of grapevine ( Vitis vinifera cv. Venus Seedless) with typical leafroll symptom originated from Xingcheng region in Liaoning Province was detected for Grapevine leafroll-associated vi-ruses (GLRaVs) by RT-PCR. The results showed that two complete sequences of major coat protein (CP) gene of GLRaV-2 (GenBank accession number FJ786017) and GLRaV-3 (FJ786016) were obtained, which indicated that the grapevine plant detected was co-infected with GLRaV-2 and GLRaV-3. The genes encoding CPm, pl9 and p24 located in 3'genomic RNA of GLRaV-2 Liaoning isolate (LN) were then cloned and se-quenced. The nucleotide sequence of GLRaV-3 CP was 942 nt, and sequence analysis showed that the identi-ties of nucleotide sequence and deduced amino acid sequence among GLRaV-3-LN and other previously repor-ted isolates including ten overseas and two domestic isolates were ranged from 89.8% to 91.8% and 94.9% to 97. 4%, respectively. The nucleotide sequences of CP, CPm (FJ786018), pl9 (FJ786019) and p24 (FJ786018) of GLRaV-2-LN were 597 nt, 672 nt, 486 nt and 618 nt in length, respectively. The relative se-quence analysis showed that the nucleotide identities of CP, CPm, pl9 and p24 among GLRaV-2-LN and oth-er isolates reported were ranged from 88.3% to 100.0% , 78.9% to 100.0% , 75.1% to 99.4% and 87.5% to 99.5% , and the identities of deduced amino acids were 92.9 to 100.0% , 89.2% to 100.0% , 73.9% to 100.0% and 89.3% to 99.0%, respectively.