基于FOX hunting system技术的蒙古柳NaCl胁迫相关基因挖掘

Mining of NaCl stress-related genes in Salix linearistipularis based on FOX hunting system technology

为挖掘蒙古柳(Salix linearistipularis)中与NaCl胁迫相关基因,以FOX hunting system技术为基础,利用酿酒酵母(Saccharomyces cerevisiae)InVscI株为试验材料,得到40多万个蒙古柳cDNAs酵母单克隆。将转基因酵母用NaCl胁迫处理后筛选得到105株抗性酵母菌,对这105株酵母菌在NaCl胁迫培养基中进行抗性检验,其中3株酵母菌对NaCl的抗性明显大于对照。通过生物信息学分析表明,3号酵母中转化的蒙古柳cDNA为全长cDNA序列,包括全部开放读码框,编码区为318 bp,编码106个氨基酸残基,序列包含DABB结构域,且与其他植物的DABB类蛋白的相似性非常高。进一步对该酵母在不同非生物胁迫处理的培养基中进行抗性分析,其对多种非生物胁迫均有明显的抗性。这一研究为进一步了解木本植物的耐盐分子机制提供试验依据。

In order to explore the genes related to NaCl stress in Salix linearistipularis,more than 400,000 yeast monoclones of S.linearistipularis cDNAs were obtained based on the FOX hunting system technology using Saccharomyces cerevisiae InVscI as material in this study.The transgenic yeast was treated with NaCl stress and screened to obtain 105 resistant yeasts,then they were carried out resistance test,in NaCl stress medium,from which three were significantly resistant to NaCl compared to the control.Bioinformatics analysis showed that the cDNA of S.linearistipularis transformed from yeast 3 is a full-length cDNA sequence,including all open reading frames,with a coding region of 318 bp,encoding 106 amino acid residues,containing the DABB structural domain,and having very high similarity to DABB-like proteins of other plants.Further results of resistance analysis of this yeast in media with different abiotic stress treatments showed significant resistance to all tested abiotic stresses.This study provides a experimental basis for further understanding of the molecular mechanisms of salt tolerance in woody plants.