酵母TPS1基因对干旱胁迫下北苍术的影响

The Effects of TPS1 Gene from Saccharomyces cerevisiae on Atractylodes chinensis under Drought Stress

目的:通过培养北苍术转TPS1抗性基因植株,探究转TPS1基因北苍术抗旱机制,提高北苍术的抗旱性。方法:以啤酒酵母基因组DNA为模板,PCR扩增、克隆TPS1基因,并构建pX6-TPS1重组载体,采用农杆菌介导法将构建的重组载体pX6-TPS1导入北苍术中,经抗生素培养基筛选获得转基因北苍术。结果:干旱胁迫后,转基因北苍术中促进根系生长的PLT1、PIN1、CRLⅠ基因表达量均显著高于野生型,其中PLT1、CRLⅠ的表达量约为野生型3倍左右,而野生型抑制根系生长的WOXⅡ、ARRⅠ基因表达量高于转基因北苍术;且干旱胁迫后,转基因北苍术根直径约为野生型的1.2倍,平均根长为野生型的2.7倍,一级侧根数目为野生型的2.4倍,转基因北苍术根部总吸收面积增加57.1%,活跃吸收面积增加57.8%,比表面积增加19.6%,根系活力指数增加30.9%。结论:转TPS1基因北苍术根系较野生型发达,有利于抵御干旱胁迫,可为抗旱育种、种质创新提供理论基础和技术指导。

Objective:To investigate the drought stress mechanism of Atractylodes chinensis through the cultivation of Atractylodes chinensis transplanting TPS1 resistance gene plant,in order to improve its drought resistance.Methods:The pX6-TPS1 recombinant vector was constructed by PCR amplification and cloning of TPS1 gene using the genomic DNA of Saccharomyces cerevisiae as template.The recombinant vector pX6-TPS1 was introduced into Atractylodes chinensis by agrobacterium-mediated method and the transgenic Atractylodes chinensis was obtained through screening of antibiotic medium.Results:After drought stress,the expressions of PLT1,PIN1 and CRLⅠgene in transgenic Atractylodes chinensis were significantly higher than that in the wild type,the expressions of gene PLT1 and CRLⅠwere about three times of the wild type;But the gene expressions of WOXⅡand ARRⅠwhich inhibited root growth were higher in wild type.The root diameter of transgenic Atractylodes chinensis was wider than 1.2 times of the wild type under drought stress,the average root length was 2.7 times than that of wild type,the number of lateral roots of the first order were more than 2.4 times of the wild type.The total absorption area of transgenic Atractylodes chinensis was increased by 57.1%,the active absorption area was increased by 57.8%,specific surface area was increased by 19.6%and the index of root activity was increased by 30.9%.Conclusion:The roots of transgenic Atractylodes chinensis are more developed than wild type,which is helpful to resist the drought stress.It can provide theoretical basis and technical guidance for drought-resistant breeding and germplasm innovation.