{"title":"芜菁花叶病毒(TuMV)反义NIb基因转基因芥菜的高抗性","authors":"Shuang Zhao, Xufeng Hao","doi":"10.1109/ICCIS.2010.267","DOIUrl":null,"url":null,"abstract":"A TuMV-NIb gene fragment was amplified from the total RNA of Mustard leaves infected with TuMV-GZ strain by RT-PCR (Reverse Transcription Polymerase Chain Reaction). This fragment was cloned into plant binary vector pBI121 in the antisense orientation and transformed into Mustard (Brassica juncea L. Linshicaoyaozi) by Agro bacterium tumefaciens mediated method. 11transgenic T0 plants were obtained. PCR (Polymerase Chain Reaction), ELISA (Enzyme-linked Immunosorbent Assay) and Southern blot analysis showed that the TuMV-Nib gene had integrated into the genome of mustard and the heredity of this gene in some lines followed the 3:1 ratio. Both T0 and T1 transgenic plants showed high levels of resistance to TuMV when artificially inoculated with TuMV. Northern blot analysis of T0 transgenic mustard revealed the integrated TuMV-NIb gene can be expressed normally.","PeriodicalId":227848,"journal":{"name":"2010 International Conference on Computational and Information Sciences","volume":"119 1","pages":"0"},"PeriodicalIF":0.0000,"publicationDate":"2010-12-17","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"1","resultStr":"{\"title\":\"High Level Resistance to TuMV (Turnip Mosaic Virus) in Transgenic Mustard with the Antisense NIb Gene of the Virus\",\"authors\":\"Shuang Zhao, Xufeng Hao\",\"doi\":\"10.1109/ICCIS.2010.267\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"A TuMV-NIb gene fragment was amplified from the total RNA of Mustard leaves infected with TuMV-GZ strain by RT-PCR (Reverse Transcription Polymerase Chain Reaction). This fragment was cloned into plant binary vector pBI121 in the antisense orientation and transformed into Mustard (Brassica juncea L. Linshicaoyaozi) by Agro bacterium tumefaciens mediated method. 11transgenic T0 plants were obtained. PCR (Polymerase Chain Reaction), ELISA (Enzyme-linked Immunosorbent Assay) and Southern blot analysis showed that the TuMV-Nib gene had integrated into the genome of mustard and the heredity of this gene in some lines followed the 3:1 ratio. Both T0 and T1 transgenic plants showed high levels of resistance to TuMV when artificially inoculated with TuMV. Northern blot analysis of T0 transgenic mustard revealed the integrated TuMV-NIb gene can be expressed normally.\",\"PeriodicalId\":227848,\"journal\":{\"name\":\"2010 International Conference on Computational and Information Sciences\",\"volume\":\"119 1\",\"pages\":\"0\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2010-12-17\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"1\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"2010 International Conference on Computational and Information Sciences\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://doi.org/10.1109/ICCIS.2010.267\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"\",\"JCRName\":\"\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"2010 International Conference on Computational and Information Sciences","FirstCategoryId":"1085","ListUrlMain":"https://doi.org/10.1109/ICCIS.2010.267","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 1
摘要
采用逆转录聚合酶链反应(RT-PCR)技术,从感染TuMV-GZ菌株的芥菜叶片总RNA中扩增出TuMV-NIb基因片段。该片段以反义定向克隆到植物二元载体pBI121中,并通过农杆菌介导法转化芥菜(Brassica juncea L. linshiicaoyaozi)。获得转基因植株11株。PCR (Polymerase Chain Reaction)、ELISA(酶联免疫吸附试验)和Southern blot分析表明,TuMV-Nib基因已整合到芥菜基因组中,部分品系的遗传比例为3:1。人工接种TuMV后,T0和T1转基因植株对TuMV均表现出较高的抗性。对T0转基因芥菜的Northern blot分析显示,整合的TuMV-NIb基因可以正常表达。
High Level Resistance to TuMV (Turnip Mosaic Virus) in Transgenic Mustard with the Antisense NIb Gene of the Virus
A TuMV-NIb gene fragment was amplified from the total RNA of Mustard leaves infected with TuMV-GZ strain by RT-PCR (Reverse Transcription Polymerase Chain Reaction). This fragment was cloned into plant binary vector pBI121 in the antisense orientation and transformed into Mustard (Brassica juncea L. Linshicaoyaozi) by Agro bacterium tumefaciens mediated method. 11transgenic T0 plants were obtained. PCR (Polymerase Chain Reaction), ELISA (Enzyme-linked Immunosorbent Assay) and Southern blot analysis showed that the TuMV-Nib gene had integrated into the genome of mustard and the heredity of this gene in some lines followed the 3:1 ratio. Both T0 and T1 transgenic plants showed high levels of resistance to TuMV when artificially inoculated with TuMV. Northern blot analysis of T0 transgenic mustard revealed the integrated TuMV-NIb gene can be expressed normally.