荞麦3个种子蛋白基因的序列和表达分析

发布时间：2018-08-04 20:45

【摘要】：【目的】阐明荞麦Fagopyrum中3个种子蛋白基因序列的特征及其在种子发育过程中的作用,为探讨荞麦种子发育过程分子机制提供理论依据。【方法】利用本地Perl程序、BLASTP、MEGA6、PLANTCARE等,分析了荞麦3个种子蛋白基因(13S球蛋白基因、11S球蛋白基因和谷蛋白基因)序列及其编码蛋白质的进化关系,预测启动子区域的顺式作用元件。通过荧光定量PCR技术分析3个基因在甜荞F.esculentum、苦荞F.tataricum和金荞F.cymosum种子发育过程中的表达情况。【结果】13S球蛋白基因含有4个外显子和3个内含子,蛋白序列长515 aa,与苦荞中的一个13S球蛋白(Gene Bank登录号:ABI32184.1)仅有6个氨基酸不同,推测二者为同一个基因,在苦荞灌浆中期表达量最高;11S球蛋白基因含有8个外显子和7个内含子,蛋白序列长914 aa,与欧洲慈姑中的一个11S球蛋白(Gene Bank登录号:ABB77213.1)亲缘关系最近,在苦荞灌浆中期表达量最高;谷蛋白基因含有3个外显子和2个内含子,蛋白序列长355 aa,与甜菜中的一个B5型谷蛋白(Gene Bank登录号:XP_010683769.1)亲缘关系最近,在甜荞灌浆中期表达量最高。3个基因的启动子区域均含有大量响应逆境和激素的顺式作用元件,以及在胚乳中表达所需的顺式作用元件。【结论】本研究获得了1个13S球蛋白基因序列、1个11S球蛋白基因序列和1个谷蛋白基因序列,这3个基因可能在胚乳中优势表达,并且参与到逆境胁迫和激素响应的过程中;灌浆中期是种子蛋白积累的关键时期,3个基因均在这一时期大量表达,这为后续进一步研究基因功能奠定了基础。
[Abstract]:[objective] to elucidate the characteristics of three seed protein gene sequences in buckwheat Fagopyrum and their roles in seed development. [methods] to provide a theoretical basis for the molecular mechanism of buckwheat seed development. The evolutionary relationships of three seed protein genes (13s globulin gene and glutenin gene) and their encoding proteins in buckwheat were analyzed and the cis-acting elements in promoter region were predicted. The expression of three genes in the seed development of F. esculentum, Tartary buckwheat F.tataricum and Tartary buckwheat F.cymosum was analyzed by fluorescence quantitative PCR. [results] the 13s globulin gene contained 4 exons and 3 introns. The protein sequence was 515aa, which was different from one 13s globulin (Gene Bank accession number: ABI32184.1 in Tartary buckwheat. They were assumed to be the same gene. The highest expression of 11s globulin gene contained 8 exons and 7 introns in the middle grain filling period of Tartary buckwheat. The protein sequence was 914aa, which was closely related to an 11s globulin (Gene Bank accession number: ABB77213.1), which was the most expressed in buckwheat during grain filling, and the glutenin gene contained three exons and two introns. The protein sequence was 355aa, which was closely related to a B5 glutenin (Gene Bank accession number: XP010683769.1), which was the most expressed in buckwheat during grain filling. The promoter regions of the three genes all contained a large number of cis-acting elements in response to stress and hormones. [conclusion] A 13s globulin gene sequence, an 11s globulin gene sequence and a glutenin gene sequence were obtained, which may be expressed in endosperm. The middle filling stage was the key stage of seed protein accumulation, and the three genes were all expressed in large quantities during this period, which laid a foundation for further study on gene function.
【作者单位】：贵州师范大学荞麦产业技术研究中心;
【基金】：国家自然科学基金(31471562;31171609) 国家现代农业产业技术体系专项资金(CARS-08-A4) 贵州省荞麦工程技术研究中心项目(黔科合农G字【2015】4003号) 贵州师范大学博士基金资助项目(11904/0516031;11904/0516026)
【分类号】：S517

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