CAJ | 학술논문

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。
2012년추계，재산동태안번가주요충식구중채집도협편퇴록，협맥안색변심적의사번가퇴록병독병화번가침염성퇴록병독병적번가양품。이용번가퇴록병독（Tomato chlorosis virus ，ToCV）적특이인물 ToCV1/ToCV2화번가침염성퇴록병독（Tomato infectious chlorosis virus ，TICV）적특이인물 TICV1/TICV2분별대양품진행확증，최후부득도이용인물 ToCV1/ToCV2확증적101 bp 적핵감산서렬，대해핵감산서렬극륭병측서。서렬비대표명，산동태안지구분리물여이등록적번가퇴록병독（ToCV）분리물상사성도재99％이상。수후，대산동태안충식구ToCV 번가분리물진행외각단백（CP）급열격단백（HSP70）서렬적확증、극륭화측서（GenBank 등록호 KC812620/KC812625），경 NCBI BLAST 비대발현，목적서렬여번가퇴록병독일본번가분리물 ToCV-Japan/Tochigi（GenBank등록호 AB513442/AB513443）상사성최고위99％，동속우모형병독속적번가퇴록병독，저시수차명학산동지구번가수도번가퇴록병독적침염。
In the autumn of 2012,tomato plants showing symptoms of interveinal chlorosis,leaf curling and nec-rotic fleck on lower leaves,similar to symptoms induced by Tomato chlorosis virus (ToCV)and Tomato infectious chlorosis virus (TICV)(two members of the genus Crinivirus in the family Closteroviridae)were collected in Tai’an, Shandong Province.Reverse transcription polymerase chain reaction (RT-PCR)was performed to test the presence of ToCV and TICVwith the specific primers ToCV1/ToCV2 and TICV1/TICV2 for ToCV and TICV,respectively.Only with ToCV primers,a 101-bp specific fragment was amplified from the symptomatic samples,and no amplification with TICV primers from any samples.Sequence analysis of the amplified fragment shared 99% nucleotide sequence identity with that of ToCV isolates registered in the NCBI.The presence of ToCV was confirmed by using the specific primer targeting the heat shock protein 70 (HSP70)gene and coat protein (CP)gene.The BLAST results showed that the target sequence from the isolates had 99% sequence identity with those from ToCV-Japan/Tochigi isolates (GenBank accession no.AB513442/AB513443),belonging to the genus Crinivirus .It is the first report that ToCV infected tomato in Shandong.