‘黄冠’梨类甜蛋白编码基因PbPR5的克隆及其表达特性分析

发布时间：2018-03-04 16:32

本文选题：‘黄冠’梨　切入点：类甜蛋白PbPR基因　出处：《果树学报》2016年02期 　论文类型：期刊论文

【摘要】：【目的】分离和克隆类甜蛋白基因Pb PR5,了解其在‘黄冠’梨不同组织、不同发育阶段以及受到非生物胁迫和植物生长调节剂处理时的表达情况。【方法】以‘黄冠’梨(Pyrus bretschneideri‘Huangguan’)组培苗为试验材料,从水杨酸(salicylic acid,SA)处理的c DNA文库中克隆获得该基因全编码区序列,命名为Pb PR5,对其进行生物信息学分析,并研究该基因在盐、聚乙二醇(EG6000)、SA和乙烯(ETH)处理后的表达情况;以12 a生‘黄冠’梨的叶片和果实为材料,探究该基因在叶片和果实不同发育阶段的表达模式。【结果】Pb PR5的c DNA序列长度为741 bp,编码224个氨基酸残基;该基因的基因组DNA序列长度为813 bp,包含1个内含子。生物信息学分析表明,Pb PR5蛋白与其他植物PR5基因编码的氨基酸序列相似性较高。实时荧光定量PCR分析表明:Pb PR5基因在‘黄冠’梨组培苗的根,茎,叶中均能表达,其中根部的表达量高于茎和叶;盐和干旱胁迫诱导了该基因表达,根和茎中的表达高峰均出现在处理后6 h,叶在盐胁迫后24 h和干旱后12 h出现表达高峰;SA和乙烯也诱导了该基因在叶片中的表达。在不同发育阶段的叶片中,老叶表达量最高,成熟叶次之,幼叶中不表达;在不同发育阶段的果实中,成熟果实中表达量最高,幼果期和膨大期果实表达量较低。【结论】Pb PR5基因在‘黄冠’梨各组织中非组成型表达,并且在‘黄冠’梨抵御生物和非生物胁迫过程中可能具有重要的生物学功能。
[Abstract]:[objective] to isolate and clone the sweet protein-like gene Pb PR5 in different tissues of 'Huangguan' pear. Expression in different developmental stages and under abiotic stress and plant growth regulator treatment. [methods] the tissue culture seedlings of 'Pyrus bretschneider Huangguan' were used as experimental materials. The full coding region of the gene, named Pb PR5, was cloned from the c DNA library treated with salicylic acid. The sequence was analyzed by bioinformatics, and the expression of the gene was studied after salt, polyethylene glycol EG6000 (SA) and ethylenediol (DNA) treatment. The expression pattern of this gene in leaves and fruits of 12-year-old 'Huangguan' pear was studied. [results] the c DNA sequence of Pb PR5 was 741bp, encoding 224 amino acid residues. The length of genomic DNA sequence of this gene is 813 BP, which contains an intron. Bioinformatics analysis shows that the amino acid sequence of PR5 gene is similar to that of other plant PR5 gene. Real-time fluorescence quantitative PCR analysis shows that the amino acid sequence of PR5 gene is similar to that of other plants. PR5 gene in the roots of 'Huangguan' pear plantlets in tissue culture, Both stem and leaf could be expressed, and the expression in root was higher than that in stem and leaf. Salt and drought stress induced the expression of the gene. The highest expression peaks in roots and stems occurred at 6 h after treatment, and the peak expression peaks of SA and ethylene in leaves at 24 h after salt stress and 12 h after drought also induced the expression of the gene in leaves. In the leaves of different developmental stages, the highest expression level was found in the old leaves. The expression of Pb PR5 gene was the highest in mature fruits and low in young and expanded fruits. [conclusion] Pb PR5 gene was expressed in all tissues of 'Huangguan' pear. And it may have important biological function in the process of resisting biological and abiotic stress.
【作者单位】：江苏徐淮地区徐州农业科学研究所;江苏徐州丰县大沙河果园;
【基金】：江苏省农业科技自主创新资金[CX(15)1023]
【分类号】：S661.2

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