ZHANG Chunhong,HU Shuying,WU Wenlong,et al.Preliminary Expression Analysis of Seedling Under Salt-tolerance in Beach Plum Using cDNA-AFLP[J].Northern Horticulture,2015,39(20):81-85.[doi:10.11937/bfyy.201520020]
滨梅幼苗耐盐相关基因差异表达的cDNA-AFLP初步分析
- Title:
- Preliminary Expression Analysis of Seedling Under Salt-tolerance in Beach Plum Using cDNA-AFLP
- Keywords:
- beach plum; cDNA-AFLP; salt-tolerance; gene expression analysis
- 文献标志码:
- A
- 摘要:
- 以生根的“滨梅5号”幼苗为试材，采用cDNA-AFLP差异显示方法研究200 mmol/L NaCl处理对基因差异表达的影响，并采用半定量RT-PCR方法验证候选基因的盐胁迫和组织表达模式。结果表明：256对选择性引物组合可扩增出4 263条可分辨的条带，且47条具有明显差异；其中24条经测序后分析表明，14条与已知片段具有较高同源性，主要涉及代谢、转录因子、胁迫相关蛋白等；获取了2条耐逆相关差异基因片段，分别与蝴蝶草天冬酰胺合成酶同源性达92%、与野草莓硫转运蛋白基因同源性达81%；与硫转运蛋白基因同源的差异基因在盐处理3 h时表达量最高，随处理时间增加而表达量逐渐下降，在各组织中以根中的表达量最高，推测其主要在根中行使功能。
- Abstract:
- Using rooted ‘Beach plum No.5′ seedlings as experimental material,differential gene expression analysis was performed under salt treatment of 200 mmol/L NaCl by using cDNA-AFLP (Amplified fragment length polymorphism).Expression patterns under salt-tolerance and in different tissues of the candidate genes were then revealed by semi-quantitative RT-PCR validation.The results showed that a total of 4 263 distinguishable bands were obtained by amplification through 256 pairs of selective primer combinations，and 47 bands were obtained with significant differences.After cloning and sequencing of 24 bands，sequence analysis of them showed that main functions of the 14 differential sequence segments comprised of metabolism，transcription factors，stress-related proteins，etc.Two fragments related to plant resistance were obtained and had the 92% homology with butterfly grass asparagine synthetase，and 81% homology with wild strawberry sulfate transporter gene respectively.The differential band homologous to the wild strawberry sulfate transporter gene was obtained and the expression level reached maximum under 3 hours treatment followed by gradual decrease with the increased processing time.Tissue expression analysis indicated that it had the highest level in roots indicating it mainly played a vital role in roots.
参考文献/References:
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备注/Memo
第一作者简介：张春红（1979-），女，博士，副研究员，现主要从事果树遗传育种等研究工作。E-mail：chzhang0714@163.com.责任作者：李维林（1966-），男，博士，研究员，现主要从事悬钩子植物资源利用等研究工作。E-mail：lwlcnbg@mail.cnbg.net.基金项目：江苏省科技支撑计划资助项目（BE2011324）；江苏省中国科学院植物研究所青年科技创新基金资助项目（青201101）。