摘要
利用同源克隆的方法从大豆品种‘科丰一号’中分离出2个RDR6基因,分别将其命名为GmRDR6a和GmRDR6b基因,并对其进行序列分析、植物组织表达以及抗逆境胁迫表达分析。结果表明:GmRDR6a基因位于大豆基因组的6号染色体上,基因全长为4 389 bp,包含一个长度为744 bp的内含子,其ORF长度为3 615 bp,编码1 204个氨基酸,相对分子质量和等电点分别为297.5×103和4.86;GmRDR6b基因位于大豆基因组的4号染色体上,基因全长为4 002 bp,包含一个长度为387 bp的内含子,其ORF长度为3 615 bp,编码1 204个氨基酸,相对分子质量和等电点分别为296.89×103和4.86;GmRDR6a和GmRDR6b都含有RDRs家族的保守序列DLDGD;2个基因在所有被检测组织中均表达,并且在花中的表达量最高;荧光定量结果表明:在大豆花叶病毒(SMV)处理下,2个基因在抗病材料‘科丰一号’中的表达量均显著高于感病材料‘南农1138-2’,在非生物胁迫下,GmRDR6a基因在盐和干旱处理下根、茎、叶组织中的表达量均提高,其中盐处理下根中的表达量最高,ABA诱导条件下出现早期响应,但是在冷害处理下该基因的表达量没有明显变化;GmRDR6b基因在盐、ABA处理下,表达量很高,冷害处理下出现早期响应,但是在干旱条件下,该基因表达趋势不明显。
Two novel RDR6 genes were identified in soybean by homologous clones,named as GmRDR6a and GmRDR6b. Sequences of the two genes were analyzed by bioinformatics softwares,and genes expression of different tissues and their reponse to biotic and abiotic stresses were performed using real-time PCR. The results showed that GmRDR6a gene was located on chromosome 6 of soybean genome,and the length of it was 4 389 bp,inclulding an intron of 744 bp. The ORF of GmRDR6a was 3 615 bp and encoded 1 204 amino acids（297. 5×103） with an isoelectric point of 4. 86. GmRDR6b gene was located on chromosome 4 of soybean genome and the length of it was 4 002 bp,contained an intron of 387 bp. The ORF of GmRDR6b was 3 615 bp and encoded 1 204 amino acids（296. 89×103） with an isoelectric point of 4. 86. The consereved motifs ＂ DLDGD＂ of RDRs gene family were identified in GmRDR6a and GmRDR6b; The expression of the two novel genes were identified in all tissues of soybean,and showed the higest expression level in flower of soybean; Real-time PCR analysis revealed that the mRNA levels of these two genes in the resistant material‘Kefeng No. 1＇were significant higher than the susuceptible material‘Nannong 1138-2＇after soybean mosaic virus（SMV） treatment. For abiotic stress,GmRDR6a showed high expression level in roots,stems and leaves under the treatments of salinity and drought,and showed the highest expression level in roots under salt stress. There was an early response after ABA treatment. There was no significant expression level difference of GmRDR6a gene under chilling stress. GmRDR6b showed higher expression level under salt stress and ABA treatment and there was an early response under chilling stress. There was no significant expression level difference of GmRDR6b gene under drought stress.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2014年第3期27-34,共8页
Journal of Nanjing Agricultural University
基金
国家大豆转基因专项(2011ZX08004-004-004)
江苏省科技支撑计划项目(BE2011305)
江苏省农业科技自主创新资金项目[cx(11)1029]
