广西植物2016，Vol.36Issue(3)：273-279,272,8.DOI:10.11931/guihaia.gxzw201401022
玉米转录因子WRKY76克隆及抗纹枯病表达模式分析
Cloning of maize WRKY76 and its expression patterns involved in resistance to maize banded sheath blight
摘要
Abstract
Maize banded leaf and sheath blight ( BLSB) which is a very destructive disease, can result in a significant yield loss and also heavily affect the quality of corn in maize production area. The expression of defense response genes could be stimulated by SA ( salicylic acid) and MeJA ( methyl jasmine) in the signal transduction pathway, and regula-ted by members of the WRKY transcription factors family. Transcription factors WRKY family members in Maize and WRKY family members which are related to resistance in Arabidopsis were collected from NCBI, and then polygenetic tree was successfully constructed. According to the result of polygenetic which was conducted by using the software CLUSTAL X and the software MEGA5.05, we concluded that the transcription factor WRKY76 was most likely involved in maize sheath blight resistance network. Whether the transcription factor WRKY76 plays a truly important role in sigjnal transduction path is still unknown. The materials used in this study were R15 ( Resistant line) and Ye478 ( Susceptible line) . R15 and Ye478 under different kinds of stress conditions were used to characterize the expression pattern of WRKY76 gene by qRT-PCR, leaf sheath samples were collected at 0, 1, 2, 4, 6, 12 and 24 h post Rhizoctonia solani AG1-IA inoculation, and 0, 1, 2, 4, 6, 12 h after SA ( salicylic acid ) and MeJA ( methyl jasmine ) stress treatment. When leaf sheath samples were inoculated with R. solani AG1-IA at jointing stage, the WRKY76 gene expres-sion was able to reach to the highest at 1 h post inoculation, and its expression was 20 and 32 times higher than control in Ye478 (Susceptible line) and R15 (Resistant line), with significant difference (P<0.05). The relative expression decreased at 2 h post inoculation. 3 times for Ye478 ( Susceptible line) while it returned to the original level for R15 ( Resistant line) . The expression pattern in R15 ( Resistant line) tended to increase along with treatment time increas-ing, while it was always higher than the susceptible line Ye478 with significant difference (P<0.05). When the maize leaf sheath was treated with SA ( salicylic acid) , the expression pattern of WRKY76 between Ye478 ( Susceptible line) and R15 ( Resistant line) were almost the same level, and its expression in R15 ( Resistant line) was higher than that of Ye478 ( Susceptible line) . For R15 ( Resistant line) , the relative expression level of WRKY76 was 33 times at 1 h post inoculation compared to the control, 46 times at 2 h post inoculation, and then decreasd to 2.5 time at 12 h post inocula-tion. However, When the maize leaf sheath was treated with MeJA, Ye478 ( Susceptible line) presents the downward trend, R15 ( Resistant line) shows a upward pattern according to the results of relative expression level at each treatment time. There was no significant difference between Ye478 ( Susceptible line) and R15 ( Resistant line) . Finally research results of this study indicated that the WRKY76 gene expression could be induced by R. solani AG1-IA, SA ( salicylic acid) and MeJA (methyl jasmine), and the WRKY76 might be involved in plant resistance signal transduction regulation pathway to maize banded leaf and sheath blight.关键词
纹枯病/转录调控因子/WRKY基因/实时荧光定量PCRKey words
maize banded leaf and sheath blight/transcription factor/WRKY76/qRT-PCR分类
农业科技引用本文复制引用
高文成,李玥,张银超,张志明,潘光堂,林海建..玉米转录因子WRKY76克隆及抗纹枯病表达模式分析[J].广西植物,2016,36(3):273-279,272,8.基金项目
国家自然科学基金(31201221)；国家“863”项目(SS2012AA100107)@@@@[Supported by the National Natural Science Foundation of China (31201221)；National High-tech R ＆D Program of China(SS2012AA100107)]。 （31201221）
