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Characterization and Functional Analysis of Differentially Expressed Genes in Bgt ‐inoculated Wheat Near‐Isogenic Lines by cDNA‐AFLP and VIGS
Author(s) -
Li Chen,
Liu Xiaoying,
Fan Baoli,
Wang Zhenying,
Peng Yongkang,
Dang Chen,
Xie Chaojie,
Liu Zhiyong
Publication year - 2014
Publication title -
crop science
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.76
H-Index - 147
eISSN - 1435-0653
pISSN - 0011-183X
DOI - 10.2135/cropsci2013.09.0639
Subject(s) - biology , blumeria graminis , powdery mildew , gene , genetics , gene expression , complementary dna , microbiology and biotechnology , plant disease resistance , botany
Differential gene expression after Blumeria graminis f. sp. tritici ( Bgt ) infection was compared in near‐isogenic lines (NILs) differing in powdery mildew resistance. RNA from Bgt ‐inoculated and Bgt ‐uninoculated (control) NILs was used for cDNA‐AFLP analysis. A total of 6644 highly reproducible fragments in the Bgt ‐inoculated NIL and 6255 fragments in the control were generated with 60 primer pairs. Forty‐two differential expression fragments (DEFs) produced reliable sequences. Sequence comparison using the National Center for Biotechnology Information database with the basic local alignment search tool (BLASTn) showed that 31 DEFs shared significant similarities with genes known to be involved in disease/defense, energy metabolism, transcription, secondary metabolism, or signal transduction. DEF38 and DEF3 were selected based on their sequence similarity to genes with known functions, and their transcriptional levels were assayed using semi‐quantitative RT‐PCR for the resistant NIL and the susceptible Jing 411 (control) sampled from the same treatment. The expression levels of DEF38 in the NIL were 1.5‐, 2.8‐, and 3.1‐ fold higher after inoculation at 8, 12, and 24 h, respectively, than those in Jing 411. DEF3 was also induced, and a significant expression change was observed in the NIL after Bgt infection. DEF38 contained a serine threonine protein kinase (STPK) domain, and was highly homologous to a stem rust‐resistant gene ( Rpg5 ) in barley. A virus‐induced gene silencing (VIGS) system was used to evaluate the function of DEF38. The result indicated that the decreased expression of DEF38 in BSMV:STPK‐DEF38‐treated seedlings compromised NIL resistance to Bgt . Multiple genes were involved in the response to Bgt infection, and disease resistance genes might play a key role in NILs.