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筛选野生种并进行转录组分析以鉴定马铃薯中响应晚疫病抗性的差异调控基因。

Screening of wild species and transcriptome profiling to identify differentially regulated genes in response to late blight resistance in potato.

作者信息

Bhatia Nisha, Tiwari Jagesh Kumar, Kumari Chandresh, Zinta Rasna, Sharma Sanjeev, Thakur Ajay Kumar, Buckseth Tanuja, Dalamu Dalamu, Singh Rajesh Kumar, Kumar Vinod

机构信息

Division of Crop Improvement, Indian Council of Agricultural Research-Central Potato Research Institute, Shimla, Himachal Pradesh, India.

School of Biotechnology, Shoolini University, Solan, Himachal Pradesh, India.

出版信息

Front Plant Sci. 2023 Jul 12;14:1212135. doi: 10.3389/fpls.2023.1212135. eCollection 2023.

Abstract

Late blight () is a serious disease of potatoes. The aim of this study was to screen wild potato species and identify differentially expressed genes (DEGs) associated with late blight resistance. Wild potato species such as PIN45 (), CPH62 (), JAM07 (), MCD24 (), PLD47 (), and cv. Kufri Bahar (control) were tested by artificial inoculation of under controlled conditions. Transcriptomes of the leaf tissues (96 h post-inoculation) were sequenced using the Illumina platform. Statistically significant ( < 0.05) DEGs were analyzed in wild species by comparison with the control, and upregulated (>2 log fold change, FC) and downregulated (<-2 log FC) genes were identified. DEGs were functionally characterized with Gene Ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Selected genes were validated by real-time PCR analysis to confirm RNA-seq results. We identified some upregulated genes associated with late blight resistance in wild species such as cytochrome P450, proline-rich protein, MYB transcription factor MYB139, ankyrin repeat-containing protein, and LRR receptor-like serine/threonine-protein kinase in PIN45; glucosyltransferase, fructose-bisphosphate aldolase, and phytophthora-inhibited protease 1 in CPH62; steroid binding protein and cysteine proteinase 3 in JAM07; glycine-rich cell wall structural protein 1 and RING finger protein in MCD24; and cysteine proteinase 3 and major latex protein in PLD47. On the other hand, downregulated genes in these species were snakin-2 and WRKY transcription factor 3 in PIN45; lichenase and phenylalanine ammonia-lyase 1 in CPH62; metallothionein and LRR receptor-like serine/threonine-protein kinase in JAM07; UDP-glucoronosyl/UDP-glucosyl transferase family protein and steroid binding protein in MCD24; and cytoplasmic small heat shock protein class I and phosphatase PLD47. Our study identified highly resistant wild potato species and underlying genes such as disease resistance, stress response, phytohormones, and transcription factors (e.g., MYB, WRKY, AP2/ERF, and AN1) associated with late blight resistance in wild potato species.

摘要

晚疫病是马铃薯的一种严重病害。本研究的目的是筛选野生马铃薯品种,并鉴定与晚疫病抗性相关的差异表达基因(DEG)。通过在可控条件下人工接种对PIN45、CPH62、JAM07、MCD24、PLD47等野生马铃薯品种以及库夫里·巴哈尔品种(对照)进行了测试。使用Illumina平台对叶片组织(接种后96小时)的转录组进行测序。通过与对照比较,对野生品种中具有统计学意义(P<0.05)的DEG进行分析,并鉴定上调(>2倍对数变化,FC)和下调(<-2倍对数FC)的基因。利用基因本体论(GO)术语和京都基因与基因组百科全书(KEGG)途径对DEG进行功能表征。通过实时PCR分析对选定基因进行验证,以确认RNA测序结果。我们在野生品种中鉴定出一些与晚疫病抗性相关的上调基因,如PIN45中的细胞色素P450、富含脯氨酸的蛋白、MYB转录因子MYB139、含锚蛋白重复序列的蛋白和LRR受体样丝氨酸/苏氨酸蛋白激酶;CPH62中的葡糖基转移酶、果糖二磷酸醛缩酶和疫霉抑制蛋白酶1;JAM07中的类固醇结合蛋白和半胱氨酸蛋白酶3;MCD24中的富含甘氨酸的细胞壁结构蛋白1和环指蛋白;以及PLD47中的半胱氨酸蛋白酶3和主要乳胶蛋白。另一方面,这些品种中的下调基因有PIN45中的蛇形蛋白-2和WRKY转录因子3;CPH62中的地衣酶和苯丙氨酸解氨酶1;JAM07中的金属硫蛋白和LRR受体样丝氨酸/苏氨酸蛋白激酶;MCD24中的UDP-葡糖醛酸基/UDP-葡糖基转移酶家族蛋白和类固醇结合蛋白;以及PLD47中的细胞质小热休克蛋白I类和磷酸酶。我们的研究鉴定出了高抗晚疫病的野生马铃薯品种以及与野生马铃薯品种晚疫病抗性相关的潜在基因,如抗病性、应激反应、植物激素和转录因子(如MYB、WRKY、AP2/ERF和AN1)。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/2f61/10368984/a5e7c9442217/fpls-14-1212135-g001.jpg

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