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Direct evidence for a new mode of plant defense against insects via a novel polygalacturonase-inhibiting protein expression strategy.
Journal of Biological Chemistry ( IF 4.0 ) Pub Date : 2020-08-14 , DOI: 10.1074/jbc.ra120.014027
Wiebke Haeger 1 , Jana Henning 1 , David G Heckel 1 , Yannick Pauchet 2 , Roy Kirsch 2
Affiliation  

Plant cell wall–associated polygalacturonase-inhibiting proteins (PGIPs) are widely distributed in the plant kingdom. They play a crucial role in plant defense against phytopathogens by inhibiting microbial polygalacturonases (PGs). PGs hydrolyze the cell wall polysaccharide pectin and are among the first enzymes to be secreted during plant infection. Recent studies demonstrated that herbivorous insects express their own PG multi-gene families, raising the question whether PGIPs also inhibit insect PGs and protect plants from herbivores. Preliminary evidence suggested that PGIPs may negatively influence larval growth of the leaf beetle Phaedon cochleariae (Coleoptera: Chrysomelidae) and identified BrPGIP3 from Chinese cabbage (Brassica rapa ssp. pekinensis) as a candidate. PGIPs are predominantly studied in planta because their heterologous expression in microbial systems is problematic and instability and aggregation of recombinant PGIPs has complicated in vitro inhibition assays. To minimize aggregate formation, we heterologously expressed BrPGIP3 fused to a glycosylphosphatidylinositol (GPI) membrane anchor, immobilizing it on the extracellular surface of insect cells. We demonstrated that BrPGIP3_GPI inhibited several P. cochleariae PGs in vitro, providing the first direct evidence of an interaction between a plant PGIP and an animal PG. Thus, plant PGIPs not only confer resistance against phytopathogens, but may also aid in defense against herbivorous beetles.

中文翻译:

通过新型多聚半乳糖醛酸酶抑制蛋白表达策略的植物防御昆虫新模式的直接证据。

植物细胞壁相关的多聚半乳糖醛酸酶抑制蛋白 (PGIPs) 广泛分布于植物界。它们通过抑制微生物多聚半乳糖醛酸酶 (PG) 在植物防御植物病原体方面发挥关键作用。PG 水解细胞壁多糖果胶,是植物感染过程中分泌的第一批酶之一。最近的研究表明,食草昆虫表达自己的 PG 多基因家族,提出了一个问题,即 PGIP 是否也抑制昆虫 PG 并保护植物免受食草动物的侵害。初步证据表明,PGIPs 可能对叶甲虫 Phaedon cochleariae (Coleoptera: Chrysomelidae) 的幼虫生长产生负面影响,并确定了来自大白菜 (Brassica rapa ssp. pekinensis) 的 BrPGIP3 作为候选物。PGIP 主要在植物中进行研究,因为它们在微生物系统中的异源表达存在问题,并且重组 PGIP 的不稳定性和聚集使体外抑制试验变得复杂。为了最大限度地减少聚集体的形成,我们异源表达了与糖基磷脂酰肌醇 (GPI) 膜锚融合的 BrPGIP3,将其固定在昆虫细胞的细胞外表面。我们证明 BrPGIP3_GPI 在体外抑制了几种 P. cochleariae PG,提供了植物 PGIP 和动物 PG 之间相互作用的第一个直接证据。因此,植物 PGIP 不仅可以抵抗植物病原体,还可以帮助防御食草甲虫。为了最大限度地减少聚集体的形成,我们异源表达了与糖基磷脂酰肌醇 (GPI) 膜锚融合的 BrPGIP3,将其固定在昆虫细胞的细胞外表面。我们证明 BrPGIP3_GPI 在体外抑制了几种 P. cochleariae PG,提供了植物 PGIP 和动物 PG 之间相互作用的第一个直接证据。因此,植物 PGIP 不仅可以抵抗植物病原体,还可以帮助防御食草甲虫。为了最大限度地减少聚集体的形成,我们异源表达了与糖基磷脂酰肌醇 (GPI) 膜锚融合的 BrPGIP3,将其固定在昆虫细胞的细胞外表面。我们证明 BrPGIP3_GPI 在体外抑制了几种 P. cochleariae PG,提供了植物 PGIP 和动物 PG 之间相互作用的第一个直接证据。因此,植物 PGIP 不仅可以抵抗植物病原体,还可以帮助防御食草甲虫。
更新日期:2020-08-14
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