汉中黑稻黄烷酮3-羟化酶基因(F3H)的遗传变异分析
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摘要
本实验通过PCR和测序拼接获得汉中地区8个黑稻品种黄烷酮3-羟化酶基因(F3H)编码区序列,在NCBI中用BLAST分析首次发现该基因位于水稻4号染色体。发现8种黑稻和普通籼稻F3H序列完全相同,与粳稻相比较,则存在4个突变位点,导致2个氨基酸突变。基于F3H序列信息的进化树表明黑稻与籼稻、粳稻、小麦、玉米的亲缘关系较近;遗传距离和同源性分析表明F3H基因较为保守,是一个古老的基因,可用于种属的鉴定分析。F3H蛋白理化性质分析发现,黑稻与其它禾本科植物存在一定差异,蛋白三维结构及相关位点预测发现粳稻和黑稻无明显差异,初步认为黑稻F3H序列变异可能与花青素的合成无关,需要在其表当量与花青苷和成合成与沉积的关系方面开展进一步研究。
The full coding sequence of F3 H was isolated from the 8 Black rice breeds in Hanzhong by PCR and sequencing.BLAST analysis found that the gene is located on the chromosome 4 of rice for the first time.The 8 black rice breeds and Indica have the same F3 H coding sequence.There were 4 mutational sites between Indica and Japonica,and resulting in 2 amino acid mutations.The phylogenetic tree indicated that black rice was very closely to Indica,Japonica,wheat,corn.The genetic distances and homology analysis showed that conserved F3 H gene was an ancient gene,and can be used to species identification.The physicochemical properties prediction of F3 H found no significant differences between Black rice and other gramineae plants.There are no significant differences in the three dimensional structure of F3 H protein and the related active site prediction of Japonica and Black rice.Therefore,it is considered that Black rice F3 H sequence variation may be unrelated to the synthesis of anthocyanins,and we need to further study on the expression level of F3 H.
The full coding sequence of F3 H was isolated from the 8 Black rice breeds in Hanzhong by PCR and sequencing.BLAST analysis found that the gene is located on the chromosome 4 of rice for the first time.The 8 black rice breeds and Indica have the same F3 H coding sequence.There were 4 mutational sites between Indica and Japonica,and resulting in 2 amino acid mutations.The phylogenetic tree indicated that black rice was very closely to Indica,Japonica,wheat,corn.The genetic distances and homology analysis showed that conserved F3 H gene was an ancient gene,and can be used to species identification.The physicochemical properties prediction of F3 H found no significant differences between Black rice and other gramineae plants.There are no significant differences in the three dimensional structure of F3 H protein and the related active site prediction of Japonica and Black rice.Therefore,it is considered that Black rice F3 H sequence variation may be unrelated to the synthesis of anthocyanins,and we need to further study on the expression level of F3 H.
引文
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[2]马挺军,任贵兴.色稻功能成分研究进展[J].中国农学通报,2010(11):61-66.
[3]Hu J P,Reddy V S,Wessler S R.The rice R gene family:Two distinct subfamilies containing several miniature inverted-repeat transposable elements[J].Plant MolBiol,2000(42):667-678.
[4]Sakamoto W,Ohmori T,Kageyama K,et al.The Purple leaf(PI)Locus of Rice:the Plw Allele has a Complex Organization and Includes Two Genes Encoding Basic Helix-Loop-Helix Proteins Involved in Anthocyanin Biosynthesis[J].Plant Cell Physiol,2001(42):982-991.
[5]马廷蕊,张金文,梁慧光,等.植物花青素合成与基因调控(英文)[J].Agricultural Science&Technology,2012(3):507-511+54.
[6]王彩霞,舒庆尧.水稻紫色种皮基因Pb的精细定位与候选基因分析[J].科学通报,2007(21):2517-2523.
[7]ZHANG Y,B0 G.R.Evluation of authentication of methodology on genoitmic DNA isolation of plant[J].Drag Evaluation,2004(4);292-297.
[8]沈芸,肖鹏,包劲松.水稻营养成分遗传育种研究进展[J].核农学报,2008(4):455-460+425.
[9]李新生,邓文辉,吴三桥,等.陕西三种特种稻米氨基酸及品质分析[J].氨基酸和生物资源,2001(4):1-3.
[10]丁锐.黑稻主要栽培品种过氧化物酶同工酶的研究[J].安徽农业科学,2005(6):947-948.
[11]王胜宝,冯志峰,李新生,等.陕西汉中特种稻产业发展现状及前景[J].新疆农业科学,2010(S2):147-150.
[12]曹小勇,李新生.黑米花色素苷类色素研究现状及展望[J].氨基酸和生物资源,2002(1):3-6.
[13]邵雅芳,徐非非,唐富福,等.水稻花青素合成相关基因的时空表达研究[J].核农学报,2013(1):9-14.
[14]李明,王玉红,李长生,等.花生黄烷酮3-羟化酶基因AhF3H的克隆和表达分析[J].山东农业科学,2013(11):1-6.
[15]石少川,高亦珂,张秀海,等.植物花青素生物合成相关基因的研究及应用[J].植物研究,2011(5):633-640.
[16]Yang Y,Zhao G,Yue W,et al.Molecular cloning and gene expression differences of the anthocyanin biosynthesis-related genes in the red/green skin color mutant of pear(Pyrus communis L.)[J].Tree Genetics&Genomes,2013(9):1351-1360.
[17]Martin C,Prescott A,Mackay S,et al.Control of anthocyanin biosynthesis in flowers of Antirrhinum majus[J].The Plant Journal,1991,1(1):37-49.
[18]Zuker A,Tzfira T,Ben-Meir H,et al.Modification of flower color and fragrance by antisense suppression of the flavanone 3-hydroxylase gene[J].Molecular Breeding,2002,9(1):33-41.
[19]Jiang F,Wang J,Jia H,et al.RNAi-mediated silencing of the flavonone 3-hydroxylase gene and its effect on flavonoid biosynthesis in strawberry fruit[J].Journal of Plant Growth Regulation,2013,32(1):182-190.
[20]Jaakola L,M(a|¨)tt(a|¨)K,Pirttil(a|¨)A M,et al.Expression of genes involved in anthocyanin biosynthesis in relation to anthocyanin,proanthocyanidin,and flavonol levels during bilberry fruit development[J].Plant Physiology,2002,130(2):729-739.