茶氨酸合成酶（Theanine synthetase, TS）基因是茶树茶氨酸代谢过程中的关键酶基因。本研究以氨基酸含量差异明显的亲本及其杂交所得F1子代为研究材料,克隆TS基因的cDNA序列,挖掘其SNPs位点,并成功将杂合SNP位点定位在遗传连锁群上。研究结果显示,通过序列比对在亲本间检测到3个SNPs,验证得到1个杂合的位点SNP735。将此位点成功转化为dCAPS标记,该标记在子代中的基因型分离比为1∶1,利用该群体已构建的茶树遗传图谱进行遗传定位,将dCAPS标记定位在连锁群LG03上,相邻标记为TM299和TM517。联合此标记及其相邻标记与游离氨基酸总含量和茶氨酸含量进行统计分析,表明具有显著的相关性。
Abstract
The theanine synthetase (TS) gene is considered as the key functional gene for the synthesis of theanine in tea plant. In this study, an F1 segregating population and its parents were used to detect single nucleotide polymorphism (SNP) in the TS gene for genetic mapping. According to the sequence alignment between the parents, three SNPs were found, and SNP735 was identified to be heterozygous in YS. SNP735 was subsequently transformed into dCAPS marker and used for genotyping in the F1 population. The results showed that the segregation ratio of alleles at this SNP locus was close to 1:1. The dCAPS marker was mapped to LG03 at a position between TM299 and TM517 in the tea genetic map. Meanwhile, significant correlations between the dCAPS marker and the content of theanine and total amino acid were identified in this study.
关键词
TS /
dCAPS标记 /
SNP /
茶氨酸 /
茶树 /
定位
Key words
TS /
dCAPS marker /
mapping /
SNP /
tea plant /
theanine
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参考文献
[1] 陈亮, 虞富莲, 杨亚军. 茶树种质资源与遗传改良[M]. 北京:中国农业科学技术出版社, 2006: 103-104.
[2] 齐桂年. 茶氨酸的研究进展[J]. 贵州茶叶, 2001(2): 15-16.
[3] 朱小兰, 陈波, 罗旭彪, 等. 高效液相色谱法测定茶叶中的茶氨酸[J]. 色谱, 2003, 21(4): 400-402.
[4] 宛晓春. 茶叶生物化学[M]. 3版. 北京: 中国农业出版社, 2003: 32-35.
[5] Okada Y, Koseki M, Chu M.Protein and cDNA sequences of two theanine synthetases from Camellia sinensis: JP, 2006254780[P]. 2006.
[6] 李娟. 安吉白茶高氨基酸性状相关基因的全长cDNA克隆及功能的初步研究[D]. 长沙: 湖南农业大学, 2011: 40-42.
[7] 陈琪, 江雪梅, 孟祥宇, 等. 茶树茶氨酸合成酶基因的酶活性验证与蛋白三维结构分析[J]. 广西植物, 2015, 35(3): 384-392.
[8] 郑德波, 杨小红, 李建生, 等. 基于SNP 标记的玉米株高及穗位高QTL定位[J]. 作物学报, 2013, 39(3): 549-556.
[9] 袁金红, 李俊华, 黄小城, 等. 基于全基因组重测序的 SNP 分析在作物基因定位中的研究进展[J]. 植物生理学报, 2015, 51(9): 1400-1404.
[10] 张学铭, 刘博, 胡云艳, 等. 白菜类作物BrFLC5与开花时间相关的dCAPs标记开发[J]. 园艺学报, 2014, 41(10): 2035-2042.
[11] 马春雷, 姚明哲, 王新超, 等. 茶树2个MYB 转录因子基因的克隆及表达分析[J]. 林业科学, 2012, 48(3): 31-37.
[12] 刘凯, 邓志英, 李青芳, 等. 利用高密度SNP遗传图谱定位小麦穗部性状基因[J]. 作物学报, 2016, 42(6): 820-831.
[13] 马建, 马小定, 赵志超, 等. 水稻抗稻瘟病基因Pi35功能性分子标记的开发及其应用[J]. 作物学报, 2015, 41(12): 1779-1790.
[14] 许家磊, 王宇, 后猛, 等. SNP检测方法的研究进展[J]. 分子植物育种, 2015, 13(2): 475-482.
[15] 束永俊, 李勇, 柏锡, 等. 大豆EST-SNP的挖掘, 鉴定及其CAPS标记的开发[J]. 作物学报, 2010, 36(4): 574-579.
[16] 张成才, 王丽鸳, 韦康, 等. 基于茶树SNP的dCAPS标记体系研究[J]. 茶叶科学, 2012, 32(6): 517-522.
[17] 葛乃蓬, 崔龙, 李汉霞, 等. 番茄抗黄化曲叶病毒基因Ty-1的双重SNP标记的开发[J]. 园艺学报, 2014, 41(8): 1583-1590.
[18] Ma J Q, Yao M Z, Ma C L, et al.Construction of a SSR-based genetic map and identification of QTLs for catechins content in tea plant (Camellia sinensis)[J]. PLOS ONE, 2014, 9(3): e93131.
[19] 陈亮, 杨亚军, 虞富莲. 茶树种质资源描述规范和数据标准[M]. 北京: 中国农业出版社, 2005: 70-71.
[20] 谭和平, 徐文平, 赵爱平, 等. GB/T 30987—2014 植物中游离氨基酸的测定[S]. 北京: 中国标准出版社, 2014: 7-24.
[21] Ma J Q, Huang L, Ma C L, et al.Large-scale SNP discovery and genotyping for constructing a high-density genetic map of tea plant using specific-locus amplified fragment sequencing (SLAF-seq)[J]. PLOS ONE, 2015, 10(6): e0128798.
[22] 马建强. 茶树高密度遗传图谱构建及重要性状QTL定位[D]. 北京: 中国农业科学院, 2013: 61-63.
[23] Sasaoka K, Kito M, Inagaki H.Studies on the biosynthesis of theanine in tea seedlings synthesis of theanine by the homogenate of tea seedlings[J]. Agricultural and Biological Chemistry, 1963, 27(6): 467-468.
[24] Sasaoka K, Kito M, Onishi Y.Some properties of the theanine synthesizing enzyme in tea seedlings[J]. Agricultural and Biological Chemistry, 1965, 29(11): 984-988.
[25] Deng W W, Ogita S, Ashihara H.Biosynthesis of theanine (γ-ethylamino-l-glutamic acid) in seedlings of Camellia sinensis[J]. Phytochemistry Letters, 2008, 1(2): 115-119.
[26] Li C F, Zhu Y, Yu Y, et al.Global transcriptome and gene regulation network for secondary metabolite biosynthesis of tea plant (Camellia sinensis)[J]. BMC Genomics, 2015, 16(1): 560.
[27] 张志鹏, 王晓玥, 黄少雄, 等. 植物功能基因单核苷酸多态性的研究进展[J]. 中国农学通报, 2016, 32(20): 25-29.
[28] Palle S R, Seeve C M, Eckert A J, et al.Association of loblolly pine xylem development gene expression with single-nucleotide polymorphisms[J]. Tree Physiology, 2013, 33(7): 763-774.
[29] Jin J Q, Yao M Z, Ma C L, et al.Natural allelic variations of TCS1 play a crucial role in caffeine biosynthesis of tea plant and its related species[J]. Plant Physiology and Biochemistry, 2016(100): 18-26.
基金
国家茶叶产业技术体系项目（CARS-023）、国家自然科学基金（31170624、31500568）
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