«Previous Article|Table of Contents|Next Article»

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:

2015年12

Page:

84-90

Research Field:

Publishing date:

Info

Title:

Orthogonal Optimization of Start Codon Targeted Polymorphism System in Curcuma wenyujin Y.H.Chen et C.Ling

Author(s):

CHEN Zhe; FENG Shang-guo; WANG Zhen; HE Ren-feng; JIANG Meng-ying; WANG Hui-zhong

College of Life and Environmental Sciences，Hangzhou Normal University，Zhejiang Provincial Key Laboratory for Genetic Improvement and Quality Control of Medicinal Plants，Hangzhou，Zhejiang 310036

Keywords:

Curcuma wenyujin Y.H.Chen et C.Ling; Start codon targeted polymorphism polymerase chain reaction（SCoT-PCR）; system optimal design

PACS:

S 567.23+9

DOI:

10.11937/bfyy.201512023

Abstract:

Taking Curcuma wenyujin Y.H.Chen et C.Ling as test material，the L₂₅(5⁶) orthogonal design was applied to optimize the five factors (template DNA，Mg²⁺，dNTPs，Taq DNA polymerase and primer concentration) of SCoT-PCR amplification system at five concentration levels，and range analysis was conducted on the PCR results.The SCoT-PCR (start codon targeted polymorphism polymerase chain reaction) system was established and optimized，in order to provide technical support for the research of the genetic diversity and molecular identification in Curcuma wenyujin Y.H.Chen et C.Ling.The results showed as follows：a total volume of 20 μL system included 0.8 μmol/L primer，0.4 mmol/L dNTPs，1.5 mmol/L Mg²⁺，0.5 U Taq DNA polymerase and 40 ng templates DNA.The effective degree order of the factors on the SCoT-PCR was as below：dNTPs>Taq DNA polymerase>primer>Mg²⁺>template DNA.The key factor affecting the SCoT-PCR system was the concentration of dNTPs. Furthermore，the optimal reaction system was used to verify with multiple varieties Curcuma wenyujin Y.H.Chen et C.Ling samples，which showed good stability，repeatability and abundent polymorphism. It was proved to be suitable for the analysis of genetic diversity and phylogenetic，the establishment of genetic maps，gene localizations and molecular mark assisted breeding researches in Curcuma wenyujin Y.H.Chen et C.Ling in future.

References:

[1]卫生部药典委员会.中国药典(一部)[S].广州:广州科技出版社,2005:144-145.[2]李洁,张岱州,高丽霞.中药郁金的现代研究概况[J].内蒙古中医药,2001,20(1):37-38.[3]汤淙淙,秦坤良,黄可新.温郁金茎叶化学成分及抗肿瘤活性[J].温州医学院学报,2007,37(2):110-113.[4]王晓慧.温郁金RAPD-PCR反应体系建立及条件优化[J].北方园艺,2008(4):226-229.[5]汤晓闯.温郁金ISSR-PCR反应体系的建立及条件优化[J].安徽农业科学,2008,36(22):9413-9415.[6]Collard B C Y,Mackill D J.Start codon targeted（SCoT）polymorphism:a simple，novel DNA marker technique for generating gene targeted markers in plants[J].Plant Mol Biol Rep,2009,27(1):86-93.[7]张君玉,郭大龙,龚莹,等.葡萄目标起始密码子多态性反应体系的优化[J].果树学报,2011,28(2):209-214.[8]陈大霞,张雪,王钰,等.应用SCoT 标记分析玄参种质资源的遗传多样性[J].中国中药杂志,2012,37(16):2368-2372.[9]何仁锋,冯尚国,陈喆,等.药用菊花SCoT-PCR反应体系的正交优化[J].核农学报,2014,28(10):1781-1789.[10]秦国新.草莓属植物SCoT分析体系的建立及优化[J].果树学报,2012,29(3):393-397.[11]苏亚春.甘蔗SCoT-PCR反应体系优化与[J].应用与环境生物学报,2012,18(5):810-818.[12]姜小凤,高燕会,童再康,等.石蒜属植物SCoT-PCR 反应体系构建及优化[J].浙江农林大学学报,2013,30(3):444-452.[13]徐旭栋,蒋瑞彬,蓝小明,等.人工栽培铁皮石斛种质资源遗传多样性的SCoT分析[J].中华中医药杂志(原中国医药学报),2013,28(7):2123-2125. [14]王慧中.分子生物学实验指南[M].北京:中国农业科学技术出版社,2009:2-3.[15]何正文,刘运生,陈立华,等.正交设计直观分析法优化PCR条件[J].湖南医科大学学报,1998,23(7):403-404.[16]王春侠,仇敬运.DNA分子标记技术在遗传育种中的应用[J].重庆科技学院学报(自然科学版),2007,9(3):17-20.[17]苏亚春,凌辉,王恒波,等.甘蔗SCoT-PCR反应体系优化与多态性引物筛选及应用[J].应用与环境生物学报,2012,18(5):810-818.[18]宣朴,邓婧,陈新,等.苦瓜ISSR 扩增条件优化的研究[J].核农学报,2006,20(3):215-217.[19]陈虎,何新华,罗聪,等.龙眼ScoT-PCR反应体系的优化[J].基因组学与应用生物学,2009,8(5):970-974.[20]侯小改,王娟,贾甜,等.牡丹SCoT分子标记正交优化及引物筛选[J].华北农学报,2011,26(5):92-96.[21]韩国辉,汪卫星,向素琼,等.多倍体枇杷ScoT分析体系的建立与优化[J].果树学报,2011,28(3):433-437.[22]夏玲,秦永华,刘成明,等.荔枝ScoT-PCR反应体系的建立及其在遗传分析中的作用[J].果树学报,2014,30(13):147-156.[23]李丕睿,蒋甲福,陈素梅,等.菊属植物ScoT分子标记技术在遗传多样性分析中的应用[J].园艺学报,2013,40(10):2015-2025.[24]龙治坚,王莹,韩国辉,等.芥菜ScoT分析体系的建立与优化[J].西南大学学报(自然科学版),2013,35(4):20-25.[25]赵瑞强,高燕会,章晓玲,等.铁皮石斛SCoT-PCR反应体系构建及优化[J].核农学报,2012,26(4):648-655.[26]Wang Y H,Xu L,Ren W M,et al.Bioactive metabolites from Chaetomium globosum L18,an endophytic fungus in the medicinal plant Curcuma wenyujin[J].Phytomedicine,2012,19(4):364-368.[27]Ahmad M G,Peter P,Zsolt P,et al.Efficiency of arbitrarily amplified dominant markers（SCOT，ISSR，RAPD）for diagnostic fingerprinting in tetraploid potato[J].American Journal of Potato Research,2011,88(3):226-237.

Memo

Memo:

-

Common functions

Last Update: 2015-08-09