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在拟南芥中诱导表达和转基因过表达韧皮部蛋白基因 AtPP2-A1 可抑制绿桃蚜 Myzus persicae 的韧皮部取食。

Harpin-induced expression and transgenic overexpression of the phloem protein gene AtPP2-A1 in Arabidopsis repress phloem feeding of the green peach aphid Myzus persicae.

机构信息

Key Laboratory of Monitoring and Management of Crop Pathogens and Insect Pests, Ministry of Agriculture of R. P. China, Nanjing Agricultural University, Nanjing 210095, PR China.

出版信息

BMC Plant Biol. 2011 Jan 13;11:11. doi: 10.1186/1471-2229-11-11.

Abstract

BACKGROUND

Treatment of plants with HrpNEa, a protein of harpin group produced by Gram-negative plant pathogenic bacteria, induces plant resistance to insect herbivores, including the green peach aphid Myzus persicae, a generalist phloem-feeding insect. Under attacks by phloem-feeding insects, plants defend themselves using the phloem-based defense mechanism, which is supposed to involve the phloem protein 2 (PP2), one of the most abundant proteins in the phloem sap. The purpose of this study was to obtain genetic evidence for the function of the Arabidopsis thaliana (Arabidopsis) PP2-encoding gene AtPP2-A1 in resistance to M. persicae when the plant was treated with HrpNEa and after the plant was transformed with AtPP2-A1.

RESULTS

The electrical penetration graph technique was used to visualize the phloem-feeding activities of apterous agamic M. persicae females on leaves of Arabidopsis plants treated with HrpNEa and an inactive protein control, respectively. A repression of phloem feeding was induced by HrpNEa in wild-type (WT) Arabidopsis but not in atpp2-a1/E/142, the plant mutant that had a defect in the AtPP2-A1 gene, the most HrpNEa-responsive of 30 AtPP2 genes. In WT rather than atpp2-a1/E/142, the deterrent effect of HrpNEa treatment on the phloem-feeding activity accompanied an enhancement of AtPP2-A1 expression. In PP2OETAt (AtPP2-A1-overexpression transgenic Arabidopsis thaliana) plants, abundant amounts of the AtPP2-A1 gene transcript were detected in different organs, including leaves, stems, calyces, and petals. All these organs had a deterrent effect on the phloem-feeding activity compared with the same organs of the transgenic control plant. When a large-scale aphid population was monitored for 24 hours, there was a significant decrease in the number of aphids that colonized leaves of HrpNEa-treated WT and PP2OETAt plants, respectively, compared with control plants.

CONCLUSIONS

The repression in phloem-feeding activities of M. persicae as a result of AtPP2-A1 overexpression, and as a deterrent effect of HrpNEa treatment in WT Arabidopsis rather than the atpp2-a1/E/142 mutant suggest that AtPP2-A1 plays a role in plant resistance to the insect, particularly at the phloem-feeding stage. The accompanied change of aphid population in leaf colonies suggests that the function of AtPP2-A1 is related to colonization of the plant.

摘要

背景

用革兰氏阴性植物病原菌产生的 harpin 组蛋白 HrpNEa 处理植物会诱导植物对包括绿桃蚜 Myzus persicae 在内的昆虫食草动物产生抗性,绿桃蚜是一种普遍吸食韧皮部的昆虫。在吸食韧皮部的昆虫的攻击下,植物使用基于韧皮部的防御机制来保护自己,该机制涉及韧皮部蛋白 2(PP2),它是韧皮部汁液中最丰富的蛋白质之一。本研究的目的是获得拟南芥(Arabidopsis)PP2 编码基因 AtPP2-A1 在植物用 HrpNEa 处理后和转化 AtPP2-A1 后的抗绿桃蚜功能的遗传证据。

结果

使用电穿孔图谱技术分别可视化了 HrpNEa 处理和非活性蛋白对照下无翅孤雌型 M. persicae 雌性在拟南芥叶片上的韧皮部取食活动。HrpNEa 诱导野生型(WT)拟南芥的韧皮部取食受到抑制,但在 AtPP2-A1 基因缺陷的植物突变体 atpp2-a1/E/142 中没有抑制,AtPP2-A1 是 30 个 AtPP2 基因中对 HrpNEa 反应最敏感的基因之一。在 WT 中而不是 atpp2-a1/E/142 中,HrpNEa 处理对韧皮部取食活动的抑制伴随着 AtPP2-A1 表达的增强。在 PP2OETAt(AtPP2-A1 过表达转基因拟南芥)植物中,不同器官(包括叶片、茎、花萼和花瓣)中均检测到大量的 AtPP2-A1 基因转录本。与转基因对照植物的相同器官相比,所有这些器官对韧皮部取食活动都有抑制作用。当监测大规模蚜虫种群 24 小时时,与对照植物相比,HrpNEa 处理的 WT 和 PP2OETAt 植物叶片上定植的蚜虫数量显著减少。

结论

由于 AtPP2-A1 过表达导致绿桃蚜的韧皮部取食活动受到抑制,以及 HrpNEa 处理 WT 拟南芥而非 atpp2-a1/E/142 突变体的抑制作用表明 AtPP2-A1 在植物对昆虫的抗性中发挥作用,特别是在韧皮部取食阶段。叶片殖民地中蚜虫种群的变化表明,AtPP2-A1 的功能与植物的定植有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/65b3/3030531/7182c0740b66/1471-2229-11-11-1.jpg

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