汉中地区不同黑稻品种查尔酮合成酶基因的遗传变异分析
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摘要
为汉中黑稻起源及其花青苷合成机制提供分子遗传依据,通过分析汉中地区8个黑稻品种和其他植物查尔酮合成酶(chalcone synthase,CHS)基因序列,利用PCR扩增8个黑稻品种查尔酮合成酶基因并测序,使用DNAMAN和MEGA5.0软件分别对水稻的CHS基因外显子序列进行比对和不同物种间的聚类。结果表明:水稻种内存在4个多态位点,8种黑稻中存在3个多态位点,比对CHS氨基酸序列,仅黑宝品种存在1个氨基酸突变。经聚类分析,8种黑稻亲缘关系最近,与籼稻聚为一类,然后与粳稻聚在一起,禾本科植物CHS同源性很高。结论:CHS是一个古老的基因,可作为种属鉴定的参考基因,CHS虽是花青苷合成的限速酶基因,但其序列的变异可能与黑稻花青苷的合成无相关性。
In order to provide a scientific basis for the origin and the synthesis mechanism of anthocyanin,the differences of chalcone synthase(CHS)gene sequenc were analyzed among O.sativaand different plants in this study.PCR amplify the CHSsequence of eight kinds of black rice in Hanzhong and sequencing,comparing the CHS mRNA sequence among O.sativa by DNAMAN and clustering between different species by MEGA5.0.Results:Three genetic variation in Hanzhong black rice was discovered and four genetic variation in O.sativa was found.Comparing the CHS amino acid sequence,one amino acid variation in HB(a kind of black rice in Hanzhong)was found.The genetic relationship rice was the closest,and the eight rice varieties were.The homology of CHS was very high in gramineae.Conclusion:CHSis an ancient gene,that can be as a reference gene for species identification.CHSis the rate-limiting enzyme,but mutation of CHS sequence may be not related to anthocyanin synthesis in rice.
In order to provide a scientific basis for the origin and the synthesis mechanism of anthocyanin,the differences of chalcone synthase(CHS)gene sequenc were analyzed among O.sativaand different plants in this study.PCR amplify the CHSsequence of eight kinds of black rice in Hanzhong and sequencing,comparing the CHS mRNA sequence among O.sativa by DNAMAN and clustering between different species by MEGA5.0.Results:Three genetic variation in Hanzhong black rice was discovered and four genetic variation in O.sativa was found.Comparing the CHS amino acid sequence,one amino acid variation in HB(a kind of black rice in Hanzhong)was found.The genetic relationship rice was the closest,and the eight rice varieties were.The homology of CHS was very high in gramineae.Conclusion:CHSis an ancient gene,that can be as a reference gene for species identification.CHSis the rate-limiting enzyme,but mutation of CHS sequence may be not related to anthocyanin synthesis in rice.
引文
[1]侯夫云,王庆美,李爱贤,等.植物花青素合成酶的研究进展[J].中国农学通报,2009,25(21):188-190.
[2]陈起萱,凌文华.黑米和红米对兔主动脉脂质斑块面积和血脂的影响[J].卫生研究,2000,29(3):170-172.
[3]谢黎虹,罗玉坤,陈能.红米和黑米的营养功效研究进展[J].西部粮油科技,2003,28(6):35-37.
[4]孙玲,张名位,池建伟,等.黑米的抗氧化性及其与黄酮和种皮色素的关系[J].营养学报,2000,22(3):246-249.
[5]李新生,吴升华.陕西黑稻资源及其开发利用[J].资源科学,1998,20(6):67-73.
[6]陈新.银杏苯丙氨酸解氨酶(PAL)的基因克隆和生物信息学分析[D].成都:成都中医药大学,2003.
[7]Ma Tingrui,Zhang Jinwen,Liang Huiguang,et al.Plant Anthocyanin Synthesis and Gene Regulation[J].Agricultural Science&Technology,2012,3:507-511,540.
[8]杨会娜,田新惠,李艳军,等.天然棕色棉查尔酮合成酶基因(GhCHS1)的克隆和实时定量表达分析[J].棉花学报,2010(1):42-48.
[9]王金玲,瞿礼嘉,陈军,等.CHS基因外显子2的进化规律及其用于植物分子系统学研究的可行性[J].科学通报,2000(9):942-950.
[10]蒙华,李成磊,吴琦等.金荞麦查尔酮合成酶基因CHS的克隆及序列分析[J].草业学报,2010(3):162-169.
[11]张明姝.菊花查尔酮合成酶基因的克隆与序列分析[D].北京:北京林业大学,2006.
[12]钟德馨,方袁梦梦,郭壮浩,等.决明查尔酮合成酶全长基因序列的克隆与分析[J].生物技术通报,2013(5):99-104.
[13]Ursula Niesbach-Kl sgen,Ellen Barzen,Jürgen Bernhardt,et al.Chalcone synthase genes in plants:A tool to study evolutionary relationships[J]Journal of Molecular Evolution,1987,26:213-1125.
[14]Zhang Y,Bog R.Evluation of authentication of methodology on genonmic DNA isolation of plant[J].Drag Evaluation,2004,4:292-297.
[15]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.
[16]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.
[17]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.
[18]张羽,李新生,冯志峰,等.陕西省有色稻资源的SSR多态性分析[J].植物遗传资源学报,2011(5):828-832.
[19]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.
[20]朱见明,李娜,张亚军,等.苜蓿黄酮的研究进展[J].草业科学,2009(9):156-162.
[21]单丽伟,汪勇,王美玲,等.大豆类黄酮生物合成关键酶CHS基因的克隆及表达分析[J].西北植物学报,2012(11):2164-2168.
[22]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(2):58-63.
[23]许锋,程水源,王燕,等.TAIL-PCR方法快速克隆银杏查尔酮合成酶基因及序列分析(英文)[J].果树学报,2007(2):237-243.
[24]赵叔灵,郭传明,陈俊愉.植物花色呈现的生物化学、分子生物学机制及其基因工程改良[J].西北植物学报,2003,23(6):1024-1035.
[25]杨丽,刘雅莉,王跃进.百合查尔酮合成酶(chs)基因的cDNA克隆与分析[J].河南农业科学,2011,40(1):124-126.
[2]陈起萱,凌文华.黑米和红米对兔主动脉脂质斑块面积和血脂的影响[J].卫生研究,2000,29(3):170-172.
[3]谢黎虹,罗玉坤,陈能.红米和黑米的营养功效研究进展[J].西部粮油科技,2003,28(6):35-37.
[4]孙玲,张名位,池建伟,等.黑米的抗氧化性及其与黄酮和种皮色素的关系[J].营养学报,2000,22(3):246-249.
[5]李新生,吴升华.陕西黑稻资源及其开发利用[J].资源科学,1998,20(6):67-73.
[6]陈新.银杏苯丙氨酸解氨酶(PAL)的基因克隆和生物信息学分析[D].成都:成都中医药大学,2003.
[7]Ma Tingrui,Zhang Jinwen,Liang Huiguang,et al.Plant Anthocyanin Synthesis and Gene Regulation[J].Agricultural Science&Technology,2012,3:507-511,540.
[8]杨会娜,田新惠,李艳军,等.天然棕色棉查尔酮合成酶基因(GhCHS1)的克隆和实时定量表达分析[J].棉花学报,2010(1):42-48.
[9]王金玲,瞿礼嘉,陈军,等.CHS基因外显子2的进化规律及其用于植物分子系统学研究的可行性[J].科学通报,2000(9):942-950.
[10]蒙华,李成磊,吴琦等.金荞麦查尔酮合成酶基因CHS的克隆及序列分析[J].草业学报,2010(3):162-169.
[11]张明姝.菊花查尔酮合成酶基因的克隆与序列分析[D].北京:北京林业大学,2006.
[12]钟德馨,方袁梦梦,郭壮浩,等.决明查尔酮合成酶全长基因序列的克隆与分析[J].生物技术通报,2013(5):99-104.
[13]Ursula Niesbach-Kl sgen,Ellen Barzen,Jürgen Bernhardt,et al.Chalcone synthase genes in plants:A tool to study evolutionary relationships[J]Journal of Molecular Evolution,1987,26:213-1125.
[14]Zhang Y,Bog R.Evluation of authentication of methodology on genonmic DNA isolation of plant[J].Drag Evaluation,2004,4:292-297.
[15]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.
[16]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.
[17]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.
[18]张羽,李新生,冯志峰,等.陕西省有色稻资源的SSR多态性分析[J].植物遗传资源学报,2011(5):828-832.
[19]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.
[20]朱见明,李娜,张亚军,等.苜蓿黄酮的研究进展[J].草业科学,2009(9):156-162.
[21]单丽伟,汪勇,王美玲,等.大豆类黄酮生物合成关键酶CHS基因的克隆及表达分析[J].西北植物学报,2012(11):2164-2168.
[22]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(2):58-63.
[23]许锋,程水源,王燕,等.TAIL-PCR方法快速克隆银杏查尔酮合成酶基因及序列分析(英文)[J].果树学报,2007(2):237-243.
[24]赵叔灵,郭传明,陈俊愉.植物花色呈现的生物化学、分子生物学机制及其基因工程改良[J].西北植物学报,2003,23(6):1024-1035.
[25]杨丽,刘雅莉,王跃进.百合查尔酮合成酶(chs)基因的cDNA克隆与分析[J].河南农业科学,2011,40(1):124-126.