Sharma Chanchal, Saripalli Gautam, Kumar Santosh, Gautam Tinku, Kumar Avneesh, Rani Sushma, Jain Neelu, Prasad Pramod, Raghuvanshi Saurabh, Jain Mukesh, Sharma J B, Prabhu K V, Sharma P K, Balyan H S, Gupta P K
Department of Genetics and Plant Breeding, Ch.Charan Singh University, Meerut, 250004, India.
Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, 110021, India.
Funct Plant Biol. 2018 Oct;45(10):1046-1064. doi: 10.1071/FP17326.
Leaf rust disease causes severe yield losses in wheat throughout the world. During the present study, high-throughput RNA-Seq analysis was used to gain insights into the role of Lr28 gene in imparting seedling leaf rust resistance in wheat. Differential expression analysis was conducted using a pair of near-isogenic lines (NILs) (HD 2329 and HD 2329+Lr28) at early (0h before inoculation (hbi), 24 and 48h after inoculation (hai)) and late stages (72, 96 and 168 hai) after inoculation with a virulent pathotype of pathogen Puccinia triticina. Expression of a large number of genes was found to be affected due to the presence/absence of Lr28. Gene ontology analysis of the differentially expressed transcripts suggested enrichment of transcripts involved in carbohydrate and amino acid metabolism, oxidative stress and hormone metabolism, in resistant and/or susceptible NILs. Genes encoding receptor like kinases (RLKs) (including ATP binding; serine threonine kinases) and other kinases were the most abundant class of genes, whose expression was affected. Genes involved in reactive oxygen species (ROS) homeostasis and several genes encoding transcription factors (TFs) (most abundant being WRKY TFs) were also identified along with some ncRNAs and histone variants. Quantitative real-time PCR was also used for validation of 39 representative selected genes. In the long term, the present study should prove useful in developing leaf rust resistant wheat cultivars through molecular breeding.
叶锈病在全球范围内导致小麦严重减产。在本研究中，采用高通量RNA测序分析来深入了解Lr28基因在赋予小麦幼苗叶锈病抗性方面的作用。使用一对近等基因系（NILs）（HD 2329和HD 2329+Lr28），在接种致病型小麦叶锈菌（Puccinia triticina）后的早期（接种前0小时（hbi）、接种后24小时和48小时（hai））和晚期（接种后72、96和168小时）进行差异表达分析。发现大量基因的表达因Lr28的存在与否而受到影响。对差异表达转录本的基因本体分析表明，在抗性和/或感病的NILs中，参与碳水化合物和氨基酸代谢、氧化应激和激素代谢的转录本有所富集。编码类受体激酶（RLKs）（包括ATP结合；丝氨酸苏氨酸激酶）和其他激酶的基因是表达受影响的最丰富的基因类别。还鉴定了参与活性氧（ROS）稳态的基因以及几个编码转录因子（TFs）的基因（最丰富的是WRKY TFs），以及一些非编码RNA和组蛋白变体。还使用定量实时PCR对39个代表性选择基因进行了验证。从长远来看，本研究应有助于通过分子育种培育抗叶锈病的小麦品种。
