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感病番茄品系中沉默蔗糖转运蛋白基因 LeHT1 会增强对番茄黄曲叶病毒的抗性,导致植株生长受抑制、病毒扩散增强和坏死。

Tomato yellow leaf curl virus infection of a resistant tomato line with a silenced sucrose transporter gene LeHT1 results in inhibition of growth, enhanced virus spread, and necrosis.

机构信息

The Otto Warburg Minerva Center for Agricultural Biotechnology and Institute of Plant Science and Genetics in Agriculture, Robert H. Smith Faculty of Agriculture, Food and Environment, The Hebrew University of Jerusalem, Rehovot 76100, Israel.

出版信息

Planta. 2010 Feb;231(3):537-48. doi: 10.1007/s00425-009-1072-6. Epub 2009 Nov 28.

Abstract

To identify genes involved in resistance of tomato to Tomato yellow leaf curl virus (TYLCV), cDNA libraries from lines resistant (R) and susceptible (S) to the virus were compared. The hexose transporter LeHT1 was found to be expressed preferentially in R tomato plants. The role of LeHT1 in the establishment of TYLCV resistance was studied in R plants where LeHT1 has been silenced using Tobacco rattle virus-induced gene silencing (TRV VIGS). Following TYLCV inoculation, LeHT1-silenced R plants showed inhibition of growth and enhanced virus accumulation and spread. In addition, a necrotic response was observed along the stem and petioles of infected LeHT1-silenced R plants, but not on infected not-silenced R plants. This response was specific of R plants since it was absent in infected LeHT1-silenced S plants. Necrosis had several characteristics of programmed cell death (PCD): DNA from necrotic tissues presented a PCD-characteristic ladder pattern, the amount of a JNK analogue increased, and production of reactive oxygen was identified by DAB staining. A similar necrotic reaction along stem and petioles was observed in LeHT1-silenced R plants infected with the DNA virus Bean dwarf mosaic virus and the RNA viruses Cucumber mosaic virus and Tobacco mosaic virus. These results constitute the first evidence for a necrotic response backing natural resistance to TYLCV in tomato, confirming that plant defense is organized in multiple layers. They demonstrate that the hexose transporter LeHT1 is essential for the expression of natural resistance against TYLCV and its expression correlates with inhibition of virus replication and movement.

摘要

为了鉴定番茄抗番茄黄曲叶病毒(TYLCV)的相关基因,比较了对病毒具有抗性(R)和易感性(S)的品系的 cDNA 文库。发现己糖转运蛋白 LeHT1 在 R 番茄植株中优先表达。使用烟草脆裂病毒诱导的基因沉默(TRV VIGS)沉默 R 植物中的 LeHT1,研究了 LeHT1 在 TYLCV 抗性建立中的作用。接种 TYLCV 后,LeHT1 沉默的 R 植物表现出生长抑制和病毒积累及扩散增强。此外,在感染的 LeHT1 沉默的 R 植物的茎和叶柄上观察到坏死反应,但在感染的未沉默的 R 植物上没有观察到。这种反应是 R 植物特有的,因为在感染的 LeHT1 沉默的 S 植物中不存在。坏死具有细胞程序性死亡(PCD)的几个特征:坏死组织的 DNA 呈现 PCD 特征梯状模式,JNK 类似物的量增加,通过 DAB 染色鉴定活性氧的产生。在感染 DNA 病毒菜豆矮化花叶病毒和 RNA 病毒黄瓜花叶病毒和烟草花叶病毒的 LeHT1 沉默的 R 植物中,也观察到沿着茎和叶柄的类似坏死反应。这些结果首次证明了番茄对 TYLCV 的坏死反应支持自然抗性,证实植物防御是多层次组织的。它们表明,己糖转运蛋白 LeHT1 是表达对 TYLCV 的天然抗性所必需的,其表达与抑制病毒复制和运动相关。

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