|Table of Contents|

Establishment of the Optimum ISSR-PCR Reaction System of Cypripedium in the Region of Changbai Mountain of China(PDF)

《北方园艺》[ISSN:1001-0009/CN:23-1247/S]

Issue:
2013年24期
Page:
92-95
Research Field:
Publishing date:

Info

Title:
Establishment of the Optimum ISSR-PCR Reaction System of Cypripedium in the Region of Changbai Mountain of China
Author(s):
SUN Ye-ying; CHEN Li-fei; LIU Shu-ying; LIU Xin; CAO Yan; LIU Hong-zhang
Colloge of Life Science,Jinlin Agricultural University，Changchun,Jilin 130118
Keywords:
Changbai mountain; Cypripedium; ISSR; reaction system
PACS:
S 682.31
DOI:
-
Abstract:
Taking Cypripedium macranthum，Cypripedium calceolus，Cypripedium ventricosum，Cypripedium shanxiense and Cypripedium guttatum of 5 Cypripedium species which distributed in Changbai mountains as test materials，the 4 factors and annealing temperatures which affect the amplification of PCR were optimized.The optimal ISSR-PCR conditions for Cypripedium were established.The results showed that containing of 30.00 ng template DNA，0.60 μmol/L primer，0.20 mmol/L dNTP，and 0.75 U Taq DNA polymerase in a volume of 25 μL was optimal.The clear，pleomorphic and repetitive amplified bands might be obtained with the above ISSR-PCR reaction conditions by 5 Cypripedium and 4 primers.It was revealed that the ISSR-PCR reaction system was suitable for researching of genetic diversity.

References:

[1]Cribb P.The Genus Cypripedium [M].Portland：Time Press,1997:126.
[2]李纪红.杓兰属和杓兰亚科生物地理学研究简况[J].郑州师范教育,2012,1（4）:42-45.
[3]李全建,王彩霞,田敏,等.扇脉杓兰AFLP反应体系的建立[J].湖南农业科学,2012(1):107-110.
[4]蔡凝枫,严宁,刘涛,等.黄花杓兰云南中甸居群遗传结构及克隆多样性的分析[J].云南植物研究,2008,30(1):69-75. 
[5]Zietkiewicz E,Rahh A,Labuda D.Genome fingerprinting by simple sequence repeat(SSR)-anchored poly-merase chain reaction amplification[J].Genomics,1994,20(2):176-183.
[6]王建波.ISSR 分子标记及其在植物遗传学研究中的应用[J].遗传,2002,24（5）:613-616.
[7]周凌瑜.小苍兰ISSR 分子标记[D].上海:上海交通大学,2008.
[8]Bornet B,Branchard M.Use of ISSR fingerprints to detect microsatellites and genetic diversity in several related Brassicataxa and Arabidopsis thaliana[J].Hereditas,2004,140：245-247.
[9]Xia T,Chen S L,Chen S Y，et al.ISSR analysis of genetic diversity of the Qinghai-Tibet Plateau endemic Rhodiola chrysanthemifolia (Crassu Laceae)[J].Biochemical Systematics and Ecology,2007,35：209-214. 
[10]Hao G,Lee D H,Lee J S，et al.A study of taxonomic relationships among species of Korean Alliumsect Sacculiferum (Alliaceae) and related species using inter-simple sequence repeat (ISSR) markers[J].Botanical Bulletin of Academia Sinica,2002,43：63-68.
[11]李敏,王尧峰,明凤.用ISSR分子标记技术分析16个蝴蝶兰品种的亲缘关系研究[J].中国农业科技导报,2010,12（1）:60-65.
[12]曹冬梅,康黎芳,王云山,等.大花萱草ISSR-PCR 最佳反应体系的建立[J].山西农业科学,2011,39（12）:1239-1242.
[13]刘小溪,荔枝林,崔光芬,等.百合ISSR-PCR反应体系的优化[J].江苏农业科学,2012,40（1）:30-33. 
[14]Li H S.ISSR markers and their application in plant genetic ariation analysis[J].Bul letin of Biology，2004，39(2)：19-21.
[15]Lu S D.Current Protocols for Molecular Biology.Second edition[M].Beijing:Peking Union Medical College Press,1999:458-463.
[16]高丽,杨波.春兰 ISSR-PCR 反应体系的优化[J].华中农业大学学报,2006,25(3):305-309.
[17]吴小丽,王树彬,曹碚生,等.辣椒 ISSR-PCR 体系优化[J].江西农业大学学报,2007,29（2）:288-291.
[18]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(4)：403-404.

Memo

Memo:
-
Last Update: 2014-12-08