Cloning and sequence analysis of the chalcone synthase gene (CHS) from Fagopyrum dibotrys

Abstract

Abstract: The conservative sequence of the chalcone synthase gene from Fagopyrum dibotrys 544 bp was isolated by homology cloning, and then the full-length DNA and cDNA ORF (open reading frame) sequences were cloned by genome-walking and RT-PCR, respectively. Sequence analysis showed that the CHS DNA of F. dibotrys was 1 650 bp, including one 462 bp intron. The coding region of cDNA was 1 188 bp, encoding 395 amino acids, designated as FdCHS and included the conserved sites of CHS such as active sites and substrate-binding pocket sites. The homology analysis of FdCHS with that of other plants showed that it was closely related to Rheum palmatum and Polygonum cuspidatum with 94% and 93% homology, respectively. The sequence accession number is GU169470 in GenBank.

CLC Number:

MENG Hua, LI Cheng-lei, WU Qi, SHAO Ji-rong, CHEN Hui. Cloning and sequence analysis of the chalcone synthase gene (CHS) from Fagopyrum dibotrys[J]. Acta Prataculturae Sinica, 2010, 19(3): 162-169.

References

[1] 李安仁. 中国植物志(25卷, 第1分册)[M]. 北京: 科学出版社, 1998: 108-111.
[2] Chan P K. Inhibition of tumor growth in vitro by the extract of Fagopyrum Cymosum[J]. Life Science, 2003, 72(16): 1851-1858.
[3] 何显忠. 金荞麦的药理作用和临床应用[J]. 时珍国医国药, 2001, 12(4): 316-317.
[4] 张政, 林汝法. 金荞麦籽粒营养成分分析[J]. 营养学报, 1999, 21(4): 480-482.
[5] Huang J X, Qu L J, Gu H Y. A preliminary study on the origin and evolution of chalcone synthase(CHS) gene in angiosperms[J]. Acta Botanic Sinica, 2004, 46(1): 10-19.
[6] 何水林, 郑金贵, 林明, 等. 植物芪类次生代谢物的功能、合成调控及基因工程研究进展[J]. 农业生物技术学报, 2004, 12(1): 102-108.
[7] 谢修志, 陈兆平, 王小菁. 非洲菊查尔酮合酶的克隆序列分析及在大肠杆菌的表达[J]. 热带亚热带植物学报, 2004, 12(5): 431-434.
[8] 杨丽, 刘雅莉, 王跃进, 等. 百合查尔酮合成酶(CHS)的克隆与分析[J]. 西北植物学报, 2006, 26(5): 933-936.
[9] 牛天敏, 马会勤, 陈尚武. 大豆查尔酮合成酶(CHS)基因的克隆、表达及其在雪莲提取液中的代谢产物分析[J]. 中国生物工程杂志, 2007, 27(2): 58-63.
[10] Xu L W, Zhou F G. Cloning,characterization and localization of CHS gene from blood orange, Citrus sinensis (L) Osbeck cv. Ruby[J]. Molecular Biology Reports, 2009, 36(7): 1983-1990.
[11] 廖海, 周嘉裕, 张超, 等. 何首乌查尔酮合成酶基因的克隆及序列分析[J]. 安徽农业科学, 2009, 37(15): 7134-7137.
[12] 吴琦, 曾子贤, 邵继荣, 等. 金荞麦内转录间隔区( ITS)的扩增及序列分析[J]. 草业学报, 2007, 16(5): 127-132.
[13] Sommer H, Saedler H. Structure of the chalcone synthase gene of Antirrhinum majus[J]. Molecular and General Genetics, 1986, 202(3): 429-434.
[14] Sharma T R. Species relationships in Fagopyrum revealed by PCR-based DNA fingerprinting[J]. Theoretical and Applied Genetics, 2002, 105(2): 306-312.
[15] 罗定泽, 侯鑫, 赵佐成. 西南地区硬枝野荞麦居群的遗传多样性研究[J]. 武汉植物学研究, 2001, 19(2): 107-112.
[16] 朱见明, 李娜, 张亚军, 等. 苜蓿黄酮的研究进展[J]. 草业科学, 2009, 26(9): 156-162.
[17] Hemleben V, Angela D, Bernhard E, et al. Characterization and structural features of a chalcone synthase mutation in a white-flowering line of Matthiola incana R. Br. (Brassicaceae)[J]. Plant Molecular Biology, 2004, 55(3): 455-465.
[18] Fukusaki E, Kawasaki K, Kajiyama S, et al. Flower color modulations of Torenia hybrida by downregulation of chalcone synthase genes with RNA interference[J]. Biotechnology, 2004, 111(3): 229-240.
[19] Cui Y, Magili J, Frederiksen R, et al. Chalcone synthase and phenylalanine ammonia lyase mRNA levels following exposure of sorghum seedling to three fungal pathogen[J]. Physiological and Molecular Plant Pathology, 1996, 49(3): 187-199.
[20] Verhoeyen M E, Bovy A, Collins G, et al. Increasing antioxidant levels in tomatoes through modification of the flavonoid biosynthetic pathway[J]. Journal of Experimental Botany, 2002, 53: 2099-2106.
[21] 燕丽萍, 夏阳, 梁惠敏, 等. 转BADH基因苜蓿T1代遗传稳定性和抗盐性研究[J]. 草业学报, 2009, 18(6): 65-71.
[22] 赵宇玮, 王英娟, 步怀宇, 等. AtNHX1基因对菊苣的转化和耐盐性研究[J]. 草业学报, 2009, 18(3): 103-109.
[23] 唐华彬, 尹迪信, 罗红军, 等. 牧草作物金荞麦的引种比较试验[J]. 贵州农业科学, 2007, 35(2): 51-53.