Dhariwal Raman, Vyas Shailendra, Bhaganagare Govindraj R, Jha Shailendra K, Khurana Jitendra P, Tyagi Akhilesh K, Prabhu Kumble V, Balyan Harindra S, Gupta Pushpendra K
Molecular Biology Laboratory, Department of Genetics and Plant Breeding, Ch. Charan Singh University, Meerut-250004, UP, India.
Interdisciplinary Centre for Plant Genomics and Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi, India.
Funct Plant Biol. 2011 Jun;38(6):479-492. doi: 10.1071/FP10246.
Genome-wide transcriptome analysis of seedling resistance to leaf rust conferred by Lr28 gene in wheat (Triticum aestivum L.) was conducted to identify differentially expressed genes during incompatible interaction. A virulent leaf rust race 77-5 was used for inoculation of resistant (HD2329+Lr28) and susceptible (HD2329 - Lr28) wheat NILs and cDNA-AFLP analyses was carried out. As many as 223 differential transcripts appeared following leaf rust inoculation; these included 122 transcripts that appeared exclusively in resistant NIL, whereas 39 transcripts appeared both in resistant and susceptible NILs. Sequence analyses of 37 transcripts, which appeared in the resistant NIL revealed that 15 transcripts had homology with genes involved in protein synthesis, signal transduction, transport, disease resistance and metabolism. The functions of remaining 22 transcripts could not be determined; these included six novel genes reported for the first time in wheat. Specific primers could be designed for 18 of the 37 transcripts, which included genes with putative and unknown functions. Quantitative real time PCR analysis was conducted using these 18 pairs of primers. A majority (13) of these transcripts appeared within 48h reaching a peak value at 96h in resistant NIL signifying their role in providing leaf rust resistance.
对小麦（Triticum aestivum L.）中由Lr28基因赋予的幼苗对叶锈病抗性进行全基因组转录组分析，以鉴定不相容互作过程中差异表达的基因。使用毒性叶锈菌小种77-5对接种抗性（HD2329+Lr28）和感病（HD2329 - Lr28）小麦近等基因系进行接种，并开展cDNA-AFLP分析。叶锈菌接种后出现了多达223个差异转录本；其中包括122个仅在抗性近等基因系中出现的转录本，而39个转录本在抗性和感病近等基因系中均出现。对在抗性近等基因系中出现的37个转录本进行序列分析，结果显示15个转录本与参与蛋白质合成、信号转导、转运、抗病性和代谢的基因具有同源性。其余22个转录本的功能无法确定；其中包括6个首次在小麦中报道的新基因。可以为37个转录本中的18个设计特异性引物，其中包括具有推定功能和未知功能的基因。使用这18对引物进行定量实时PCR分析。这些转录本中的大多数（13个）在48小时内出现，在抗性近等基因系中于96小时达到峰值，表明它们在提供叶锈病抗性方面发挥作用。
