小麦RING型E3泛素连接酶基因TaSDIR1-D克隆与功能分析
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1山西大学生物工程学院,太原 030000;,2中国农业科学院作物科学研究所,北京 100081,2中国农业科学院作物科学研究所,北京 100081,2中国农业科学院作物科学研究所,北京 100081,1山西大学生物工程学院,太原 030000;,2中国农业科学院作物科学研究所,北京 100081

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自然科学基金(31271720);中国农业科学院科技创新工程协同创新任务(CAAS-XTCX2016019)


Cloning and Functional Analysis of a RING-type E3 Ubiquitin Ligase Gene TaSDIR1-D in Wheat
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1College of Bioengineering, University of Shanxi, Taiyuan 030000;,2Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081,2Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081,2Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081,1College of Bioengineering, University of Shanxi, Taiyuan 030000;,2Institute of Crop Science, Chinese Academy of Agricultural Sciences, Beijing 100081

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    摘要:

    具有RING结构域的E3泛素连接酶SDIR1(SALT-AND DROUGHT INDUCED REALLY INTREASTING NEW GENE FINGER1)在植物信号调节通路中发挥着重要的作用。本研究克隆得到小麦TaSDIR1-D基因,基因组序列全长4070 bp,其中编码区上游为1443 bp,编码区为2352 bp,3′端非编码区(UTR)为275 bp,该基因编码区cDNA序列长度为849 bp,编码282个氨基酸,包括2个跨膜结构域和1个保守的RING型finger结构域。以野生近缘种的二倍体、四倍体和六倍体普通小麦及一套由中国春为背景的缺四体为材料,将基因定位在染色体4D上;小麦抽穗期TaSDIR1-D在旗叶中的表达量最高;在NaCl、ABA、PEG及4℃非生物胁迫诱导下,小麦TaSDIR1-D均上调表达,可能参与植物抗逆调节通路;通过检测32份多态性较高的小麦材料TaSDIR1-D基因序列多态性,共检测到2个SNP,基因的编码区和上游序列中各存在1个核苷酸变异,其中编码区的核苷酸变异位点(G/A)位于第4外显子上,为非同义突变,使精氨酸(CGC)变为组氨酸(CAC)。2个SNP组成两种单倍型,根据-583 bp处的SNP(T/C)设计分子标记SNP-583,扫描自然群体262份材料的基因型,将其分为2种单倍型,分别与千粒重、倒二节长和穗长显著相关或极显著相关,单倍型Hap-4D-2为提高千粒重的优异单倍型。研究结果为小麦分子育种提供了理论依据和基因资源。

    Abstract:

    A RING-type E3 ubiquitin ligase SDIR1 (SALT-AND DROUGHT INDUCED REALLY INTREASTING NEW GENE FINGER1), which plays a pivotal role in plants signaling pathway. TaSDIR1-4D was isolated from genome D of wheat (Triticum aestivum L.). The full length of TaSDIR1-4D was 4,070 bp, including a 1443 bp upstream fragment of the coding region, 2352 bp coding region, 275 bp 3' UTR.The gene cDNA sequence length of the coding region was 849 bp, which encoded 282 amino acids, including two transmembrane domains and a relatively conserved RING finger domain. TaSDIR1-D was located on the chromosome 4D by using wheat wild relatives and a set of nullisomic-tetrasomic lines of Chinese Spring. TaSDIR1-D showed the highest expression level in flag leaves during heading stage. TaSDIR1-D was up-regulated by NaCl, ABA, PEG and 4℃ treatments, which suggested that TaSDIR1-D may be involved in plant abiotic stress signaling pathway. By detecting the polymorphism of TaSDIR1-D gene sequence in 32 hexaploid wheat accessions, two SNPs were detected in the full-length sequence. One was identified in the promoter region, another in the fourth exon (G/A), which led to amino acid Arg (CGC) change into His (CAC). In a natural population consisted of 262 accessions, two haplotypes of TaSDIR1-D were detected by SNP-583, a SNP marker developed from the SNP (T/C) at -583 bp, which were significantly or very significantly correlated with 1000-grain weight, length of penultimate node and spike length. Hap-4A-2 is considered as a potential superior haplotype for increasing 1000-grain weight. The results provide a theoretical basis and genetic resources for wheat molecular breeding.

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王瑞同,王景一,毛新国,等.小麦RING型E3泛素连接酶基因TaSDIR1-D克隆与功能分析[J].植物遗传资源学报,2018,19(5):951-958.

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  • 收稿日期:2018-03-09
  • 最后修改日期:2018-04-10
  • 录用日期:2018-04-13
  • 在线发布日期: 2018-06-27
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