Transcriptome analysis of Phytophthora cactorum infecting strawberry identified RXLR effectors that induce cell death when transiently expressed in Nicotiana benthamiana

Bikal Ghimire, A. Gogoi, Mandeep Poudel, Arne Stensvand, M. Brurberg
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Abstract

Phytophthora cactorum is a plant pathogenic oomycete that causes crown rot in strawberry leading to significant economic losses every year. To invade the host, P. cactorum secretes an arsenal of effectors that can manipulate host physiology and impair its defense system promoting infection. A transcriptome analysis was conducted on a susceptible wild strawberry genotype (Fragaria vesca) 48 hours post inoculation with P. cactorum to identify effectors expressed during the early infection stage. The analysis revealed 4,668 P. cactorum genes expressed during infection of F. vesca. A total of 539 secreted proteins encoded by transcripts were identified, including 120 carbohydrate-active enzymes, 40 RXLRs, 23 proteolytic enzymes, nine elicitins, seven cysteine rich proteins, seven necrosis inducing proteins and 216 hypothetical proteins with unknown function. Twenty of the 40 RXLR effector candidates were transiently expressed in Nicotiana benthamiana using agroinfiltration and five previously unreported RXLR effector genes (Pc741, Pc8318, Pc10890, Pc20813, and Pc22290) triggered cell death when transiently expressed. The identified cell death inducing RXLR effectors showed 31–66% identity to known RXLR effectors in different Phytophthora species having roles in pathogenicity including both activation and suppression of defense response in the host. Furthermore, homology analysis revealed that these cell death inducing RXLR effectors were highly conserved (82 - 100% identity) across 23 different strains of P. cactorum originating from apple or strawberry.
对感染草莓的仙客来病菌进行转录组分析,确定了在烟草中瞬时表达可诱导细胞死亡的 RXLR 效应子
噬菌体是一种植物病原卵菌,每年都会导致草莓冠腐病,造成重大经济损失。为了入侵宿主,P. cactorum 会分泌一系列效应物,这些效应物可以操纵宿主的生理机能,损害其防御系统,从而促进感染。对易感野生草莓基因型(Fragaria vesca)在接种 P. cactorum 48 小时后进行了转录组分析,以确定在早期感染阶段表达的效应物。分析结果显示,4668 个 P. cactorum 基因在 F. vesca 感染期间表达。共鉴定出 539 个由转录本编码的分泌蛋白,包括 120 个碳水化合物活性酶、40 个 RXLR、23 个蛋白水解酶、9 个激发素、7 个富半胱氨酸蛋白、7 个坏死诱导蛋白和 216 个功能未知的假定蛋白。利用农渗技术在烟草中瞬时表达了 40 个候选 RXLR 效应基因中的 20 个,其中 5 个以前未报道过的 RXLR 效应基因(Pc741、Pc8318、Pc10890、Pc20813 和 Pc22290)在瞬时表达时会引发细胞死亡。鉴定出的诱导细胞死亡的 RXLR 效应子与不同疫霉菌种中已知的 RXLR 效应子有 31-66% 的相似性,这些效应子在致病性中的作用包括激活和抑制宿主的防御反应。此外,同源性分析表明,这些诱导细胞死亡的 RXLR 效应子在源自苹果或草莓的 23 个不同菌株中高度保守(82 - 100%相同)。
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