Identification of Avramr1 from Phytophthora infestans using long read and cDNA pathogen-enrichment sequencing (PenSeq).,Molecular Plant Pathology

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Identification of Avramr1 from Phytophthora infestans using long read and cDNA pathogen-enrichment sequencing (PenSeq).
Molecular Plant Pathology ( IF 4.8 ) Pub Date : 2020-09-15 , DOI: 10.1111/mpp.12987
Xiao Lin ₁ , Tianqiao Song ₁ , Sebastian Fairhead ₁ , Kamil Witek ₁ , Agathe Jouet ₁ , Florian Jupe ₁ , Agnieszka I Witek ₁ , Hari S Karki ₁ , Vivianne G A A Vleeshouwers ₂ , Ingo Hein _{3,

4} , Jonathan D G Jones ₁

Affiliation

Potato late blight, caused by the oomycete pathogen Phytophthora infestans, significantly hampers potato production. Recently, a new Resistance to Phytophthora infestans (Rpi) gene, Rpi‐amr1, was cloned from a wild Solanum species, Solanum americanum. Identification of the corresponding recognized effector (Avirulence or Avr) genes from P. infestans is key to elucidating their naturally occurring sequence variation, which in turn informs the potential durability of the cognate late blight resistance. To identify the P. infestans effector recognized by Rpi‐amr1, we screened available RXLR effector libraries and used long read and cDNA pathogen‐enrichment sequencing (PenSeq) on four P. infestans isolates to explore the untested effectors. Using single‐molecule real‐time sequencing (SMRT) and cDNA PenSeq, we identified 47 highly expressed effectors from P. infestans, including PITG_07569, which triggers a highly specific cell death response when transiently coexpressed with Rpi‐amr1 in Nicotiana benthamiana, suggesting that PITG_07569 is Avramr1. Here we demonstrate that long read and cDNA PenSeq enables the identification of full‐length RXLR effector families and their expression profile. This study has revealed key insights into the evolution and polymorphism of a complex RXLR effector family that is associated with the recognition by Rpi‐amr1.

中文翻译：

使用长读长和 cDNA 病原体富集测序 (PenSeq) 鉴定致病疫霉中的 Avramr1。

马铃薯晚疫病由卵菌病原体致病疫霉引起，严重阻碍马铃薯生产。最近，从野生茄属植物美洲茄中克隆了一种新的抗致病疫霉( Rpi ) 基因Rpi-amr1 。从致病疫霉中鉴定相应的公认效应基因（无毒力或Avr ）是阐明其自然发生的序列变异的关键，这反过来又揭示了同源晚疫病抗性的潜在持久性。为了鉴定Rpi-amr1识别的致病疫霉效应子，我们筛选了可用的 RXLR 效应子文库，并对四种致病疫霉分离株使用长读长和 cDNA 病原体富集测序 (PenSeq)，以探索未经测试的效应子。使用单分子实时测序 (SMRT) 和 cDNA PenSeq，我们鉴定了 47 个来自致病疫霉的高表达效应子，其中包括 PITG_07569，当与本塞姆氏烟草中的Rpi-amr1瞬时共表达时，它会触发高度特异性的细胞死亡反应，这表明PITG_07569是Avramr1 。在这里，我们证明长读长和 cDNA PenSeq 能够识别全长 RXLR 效应子家族及其表达谱。这项研究揭示了与Rpi-amr1识别相关的复杂 RXLR 效应家族的进化和多态性的重要见解。

更新日期：2020-10-12

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