烟草科技
煙草科技
연초과기
TOBACCO SCIENCE & TECHNOLOGY
2015年
1期
21-25,53
,共6页
朱丹%安梦楠%赵艳琴%赵秀香%吴元华
硃丹%安夢楠%趙豔琴%趙秀香%吳元華
주단%안몽남%조염금%조수향%오원화
烟草%抗PVYN突变株%eIF4E基因%克隆%表达
煙草%抗PVYN突變株%eIF4E基因%剋隆%錶達
연초%항PVYN돌변주%eIF4E기인%극륭%표체
Tobacco%Mutant resistant to PVYN%eIF4E gene%Cloning%Expression
为了明确抗马铃薯Y病毒病的相关基因、进一步揭示其抗病机制,以烟草高抗马铃薯Y病毒脉坏死株系(Potato virus Y vein necrosis strain,PVYN)突变株SN01和SN02为试验材料,研究了突变株的抗病相关基因翻译起始因子4E(eIF4E)在寄主中的抗病作用。结果表明:从抗病突变株SN01和SN02中克隆到目的全长基因,分别命名为eIF4E01与eIF4E02,其开放阅读框ORF长度均为669 bp,编码222个氨基酸;利用MAGA4.1和CLUSTAL软件进行序列同源性比对分析表明,eIF4E01和eIF4E02与已报道的烟草eIF4E(GenBank:AY702653.1)基因序列相似度均达到99%,其开放阅读框序列完全一致。并与甜椒、番茄、拟南芥等物种的eIF4E家族基因也具有高度同源性;Real-time PCR结果表明,在PVYN侵染初期(接种后0~9 d),突变株SN01和SN02中的eIF4E基因表达量明显低于其感病亲本NC89和K326。说明这两个抗病突变株中的eIF4E基因的抑制表达与烟草对PVYN的抗性密切相关。
為瞭明確抗馬鈴藷Y病毒病的相關基因、進一步揭示其抗病機製,以煙草高抗馬鈴藷Y病毒脈壞死株繫(Potato virus Y vein necrosis strain,PVYN)突變株SN01和SN02為試驗材料,研究瞭突變株的抗病相關基因翻譯起始因子4E(eIF4E)在寄主中的抗病作用。結果錶明:從抗病突變株SN01和SN02中剋隆到目的全長基因,分彆命名為eIF4E01與eIF4E02,其開放閱讀框ORF長度均為669 bp,編碼222箇氨基痠;利用MAGA4.1和CLUSTAL軟件進行序列同源性比對分析錶明,eIF4E01和eIF4E02與已報道的煙草eIF4E(GenBank:AY702653.1)基因序列相似度均達到99%,其開放閱讀框序列完全一緻。併與甜椒、番茄、擬南芥等物種的eIF4E傢族基因也具有高度同源性;Real-time PCR結果錶明,在PVYN侵染初期(接種後0~9 d),突變株SN01和SN02中的eIF4E基因錶達量明顯低于其感病親本NC89和K326。說明這兩箇抗病突變株中的eIF4E基因的抑製錶達與煙草對PVYN的抗性密切相關。
위료명학항마령서Y병독병적상관기인、진일보게시기항병궤제,이연초고항마령서Y병독맥배사주계(Potato virus Y vein necrosis strain,PVYN)돌변주SN01화SN02위시험재료,연구료돌변주적항병상관기인번역기시인자4E(eIF4E)재기주중적항병작용。결과표명:종항병돌변주SN01화SN02중극륭도목적전장기인,분별명명위eIF4E01여eIF4E02,기개방열독광ORF장도균위669 bp,편마222개안기산;이용MAGA4.1화CLUSTAL연건진행서렬동원성비대분석표명,eIF4E01화eIF4E02여이보도적연초eIF4E(GenBank:AY702653.1)기인서렬상사도균체도99%,기개방열독광서렬완전일치。병여첨초、번가、의남개등물충적eIF4E가족기인야구유고도동원성;Real-time PCR결과표명,재PVYN침염초기(접충후0~9 d),돌변주SN01화SN02중적eIF4E기인표체량명현저우기감병친본NC89화K326。설명저량개항병돌변주중적eIF4E기인적억제표체여연초대PVYN적항성밀절상관。
In order to explore PVY resistance-related gene and further reveal its resistance mechanism, the tobacco mutants SN01 and SN02 highly resistant to potato virus Y vein necrosis strain (PVYN) were experimented to study the role of translation initiation factor eIF4E of PVY resistance-related gene in the mutants. The results showed that full length target genes were successfully cloned from the mutants SN01 and SN02. The genes were referred to as eIF4E01 and eIF4E02, respectively; their open reading frame (ORF) was 669 bp in length and encoded a protein of 222 amino acids. The homology analysis by MEGA 4.1 and CLUSTAL showed that these genes possessed 99% sequence similarities to and exactly the same ORF sequence as reported eIF4E in tobacco (GenBank: AY702653.1), they were highly homologous to the eIF4E family genes in Capsicum annuum, Lycopersicon esculentum and Arabidopsis thaliana. Real-time PCR results indicated that the expression level of eIF4E in the mutants was obviously lower than that in their susceptible parents NC89 and K326 at the early stage of PVYN infection (0-9 days after inoculation), it indicated that the suppression of eIF4E expression in the two mutants was closely related to the resistance of tobacco to PVYN.