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在表达病毒复制酶(Nlb)基因的转基因豌豆中对豌豆种传花叶马铃薯Y病毒抗性的特异性。

Specificity of resistance to pea seed-borne mosaic potyvirus in transgenic peas expressing the viral replicase (Nlb) gene.

作者信息

Jones A L, Johansen I E, Bean S J, Bach I, Maule A J

机构信息

John Innes Centre, Norwich Research Park, Colney, UK.

出版信息

J Gen Virol. 1998 Dec;79 ( Pt 12):3129-37. doi: 10.1099/0022-1317-79-12-3129.

Abstract

Transgenic pea lines carrying the replicase (NIb) gene of pea seed-borne mosaic potyvirus (PSbMV) were generated and used in experiments to determine the effectiveness of induced resistance upon heterologous isolates. Three pea lines showed inducible resistance in which an initial infection by the homologous isolate (PSbMV-DPD1) was followed by a highly resistant state. Resistance was observed in plants in either the homozygous or hemizygous condition and resulted in no overall yield loss despite the initial infection. Resistance was associated with a loss of both viral and transgene RNA, which is indicative of a mechanism based upon post-transcriptional gene silencing. There was no correlation between the steady-state levels of transgene RNA and ability of the plants to show resistance. To test the specificity of the resistance, plants were also inoculated with the most distantly related sequenced PSbMV isolate, NY. PSbMV-NY varied between experiments in its ability to induce resistance, suggesting that the sequence identity in the NIb gene is borderline for the specificity required for triggering gene silencing. Upon challenge inoculation of virus-free recovered leaves, the specificity of the induced resistance varied between the two isolates and indicated that the virus and transgene additively determined the resistant state. These results suggest that the sequence requirements for triggering gene silencing may differ from those involved in the degradation process.

摘要

构建了携带豌豆种传花叶马铃薯Y病毒(PSbMV)复制酶(NIb)基因的转基因豌豆品系,并用于实验以确定对异源分离株诱导抗性的有效性。三个豌豆品系表现出可诱导抗性,其中同源分离株(PSbMV-DPD1)的初始感染之后是高度抗性状态。在纯合或半合子状态的植物中均观察到抗性,并且尽管有初始感染,但总体产量没有损失。抗性与病毒RNA和转基因RNA的丧失有关,这表明是一种基于转录后基因沉默的机制。转基因RNA的稳态水平与植物表现抗性的能力之间没有相关性。为了测试抗性的特异性,还对植物接种了亲缘关系最远的已测序PSbMV分离株NY。PSbMV-NY在不同实验中诱导抗性的能力有所不同,这表明NIb基因中的序列同一性对于触发基因沉默所需的特异性处于临界状态。对接种无病毒后恢复的叶片进行挑战接种时,两种分离株诱导抗性的特异性有所不同,表明病毒和转基因共同决定了抗性状态。这些结果表明,触发基因沉默的序列要求可能与参与降解过程的序列要求不同。

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