ZHAO Li-hua.Establishment of the Optimized AFLP Analysis System for Pomegranate[J].Northern Horticulture,2012,36(21):79-82.
石榴AFLP反应体系的建立及优化
- Title:
- Establishment of the Optimized AFLP Analysis System for Pomegranate
- 文章编号:
- 1001-0009（2012）21-0079-04
- Keywords:
- pomegranates; AFLP; system; optimized
- 分类号:
- S 665.4
- 文献标志码:
- A
- 摘要:
- 以提取的‘粉红牡丹’石榴叶片基因组DNA为材料，对影响AFLP反应体系的Mse I/EcoRⅠ酶切时间、预扩增模板DNA稀释倍数及程序和选择性扩增模板DNA稀释倍数等关键因素进行优化。结果表明：在石榴AFLP反应体系为20 μL双酶切反应体系中，基因组DNA为300 ng的最佳酶切时间为6 h；以不稀释DNA连接产物为预扩增模板，94℃为预扩增反应程序的起始解链温度；以稀释20倍预扩产物DNA为选扩模板。
- Abstract:
- Taking genome DNA of pomegranate ‘Fenhongmudan’ as material，AFLP reaction system was optimized from time of Mse I/EcoRⅠ restriction enzyme digest system，template DNA dilution multiple and program of pre-amplification，template DNA dilution multiple of selective amplification.The results indicated that 300 ng genomic DNA was digested completely with 6 hours，the products of ligation was not diluted；starting melting temperature was 94℃ for pre-amplification；the products of pre-amplification was diluted for 20 times.
参考文献/References:
[1]张玉星.果树栽培学各论[M].1版.北京:中国农业出版社,2008:78-86. [2]汪小飞.石榴品种分类研究[D].南京:南京林业大学,2007:20-49. [3]Stover E，Mercure E W.The pomegranate：a new look at the fruit of paradise[J].Hort Science，2007,42(5):1088-1092. [4]卢龙斗,刘素霞,邓传良,等.RAPD技术在石榴品种分类上的应用[J].果树学报,2007，24(5):634-639. [5]Ghobadi S，Khoushkhoui M，Tabatabaei B E S.Phylogenetic relationships among some iranin ponegranate accessions revealed by Inter-simple Sequence Repeat(ISSR) markers[J].Iranlan Journal of Horticultural Science and Technoogy Fall,2005，6(3):111-120. [6]Narzary D，Rana T S，Ranade S A.Genetic diversity in inter-simple sequence repeat profiles across natural populations of Indian pomegranate(Punica granatum L.)[J].Plant Biology,2010,12(5):806-813. [7]曹仪植.植物分子生物学[M].1版.北京:教育出版社,2004:142-159. [8]周延清.DNA分子标记技术在植物研究中的应用[M].1版.北京:化学工业出版社,2005:2-179. [9]赵丽华,王先磊.成熟石榴叶片DNA提取方法研究[J].安徽农业科学,2009,37(31):15141-15143. [10]翟文慧,贾春枫,周莹,等.十字花科蔬菜黑腐病菌(Xanthomonas campestris?cv. campestris)AFLP分析体系的建立及优化[J].中国农学通报,2011,27(22):168-172. [11]Aitken K S，Li J C，Jackson P，et al.AFLP analysis of genetic diversity within saccharum officinarum and comparison with sugarcane cultivars[J].Aust J Agric Res,2006，57：1167-1184. [12]Li W，Xing S Y，Yang K Q，et al.Genetic relationships of ornamental cultivars of Ginkgo biloba analyzed by AFLP techniques[J].Acta Genetica Sinica,2006,33(11)：1020-1026. [13]Byun S Q，Fang Q，Zhou H，et al.An effective method for silver-staining DNA in large numbers of polyacrylamide gels[J].Anal Biochem，2009，38(5)：174-175. [14]Yu X Y，Li M，Liu Z，et al.A novel gene，screened by cDNA-AFLP approach，contributes to lowering the acidity of fruit in apple[J].Plant Physiology and Biochemistry，2007，45:139-145. [15]张正银,刘波洋,陈放,等.麻疯树AFLP体系的建立与优化[J].应用与环境生物学报,2009,15(2):280-283. [16]臧德奎,陈红,郑林,等.木瓜属优良品种亲缘关系的AFLP分析[J].林业科学,2009,45(8):39-43. [17]赵志常,胡桂兵,刘运春,等.番荔枝DNA的提取和AFLP体系的建立[J].北方园艺,2009(10):44-47. [18]明军,张启翔,晏小兰,等.梅花基因组AFLP银染反应体系的建立和优化[J].北京林业大学学报,2003,25(3):17-21. [19]田晓俊,温强,汪信东,等.闽楠、红楠AFLP反应体系建立[J].林业科技开发,2009,23(3):38-42. [20]欧立军,黄园,王俞人,等.天门冬AFLP反应体系的建立及优化[J].中国农学通报,2011,27(8):87-90. [21]Yan Z，Denneboom C，Hattendorf A.Construction of an integrated map of rose with AFLP,SSR,PK,RGA,RFLP,SCAR and morphological markers[J].Theor Appl Genet,2005,110:766-777. [22]刘欢.燕麦AFLP标记的遗传多样性分析[D].兰州:甘肃农业大学,2007. [23]李元军,唐美玲,于青,等.富士苹果AFLP体系的优化及其在鉴定早熟芽变中的应用[J].园艺学报,2009,36(3):327-332.
相似文献/References:
[1]张玉平,姜明,鲁绍伟,等.黑莓亲缘关系的AFLP分析[J].北方园艺,2013,37(21):116.
ZHANG Yu-ping,JIANG Ming,LU Shao-wei,et al.Genetic Relationship Analysis of Blackberries by AFLP[J].Northern Horticulture,2013,37(21):116.
[2]黄颖宏,李康,严斌,等.水杨酸对石榴扦插苗的影响[J].北方园艺,2012,36(08):86.
HUANG Ying-hong,LI Kang,YAN Bin,et al.Effects of Salicylic Acid on the Cutting Seedling of Pomegranate[J].Northern Horticulture,2012,36(21):86.
[3]马丽,王玉海,明东风.石榴种质资源过氧化物酶同工酶的亲缘关系分析[J].北方园艺,2012,36(19):124.
MA Li,WANG Yu-hai,MING Dong-feng.Analysis of Genetic Relationship for Punica granatum L.Germplasm Based on Peroxidase Isozyme[J].Northern Horticulture,2012,36(21):124.
[4]田忠景,康美玲,王秀文.石榴叶片蛋白提取方法研究[J].北方园艺,2012,36(19):137.
TIAN Zhong-jing,KANG Mei-ling,WANG Xiu-wen.Study on Protein Extraction Methods of Pomegranate Leaves[J].Northern Horticulture,2012,36(21):137.
[5]刘兴满,葛金涛,吴秋月.石榴分子标记的研究进展[J].北方园艺,2015,39(08):194.[doi:10.11937/bfyy.201508051]
LIU Xing-man,GE Jin-tao,WU Qiu-yue.Research Progress on Molecular Marker of Pomegranate (Punica granatum L.)[J].Northern Horticulture,2015,39(21):194.[doi:10.11937/bfyy.201508051]
[6]戚 飞,刘 冠,李景富.番茄果实有机酸总含量的QTL分析[J].北方园艺,2015,39(05):109.[doi:10.11937/bfyy.201505033]
QI Fei,LIU Guan,LI Jing-fu.QTL Analysis for Total Organic Acid Content in Tomato Fruit[J].Northern Horticulture,2015,39(21):109.[doi:10.11937/bfyy.201505033]
[7]赵 丽 华.石榴ISSR-PCR反应体系的正交设计优化[J].北方园艺,2010,34(19):0.[doi:10.11937/bfyy.201019058]
ZHAO Li -hua.Optimization of ISSR -PCR Reaction System for Pomegranate in Orthogonal Design[J].Northern Horticulture,2010,34(21):0.[doi:10.11937/bfyy.201019058]
[8]陈芸,高原青,王继莲,等.不同保存方法对石榴成熟叶片基因组DNA提取效果的影响[J].北方园艺,2016,40(08):100.[doi:10.11937/bfyy.201608028]
CHEN Yun,GAO Yuanqing,WANG Jilian,et al.Effect of Different Preservation Methods on Genomic DNA Extraction of Pomegranate[J].Northern Horticulture,2016,40(21):100.[doi:10.11937/bfyy.201608028]
[9]冯立娟,尹燕雷,杨雪梅,等.石榴果皮DAHPS基因的克隆与生物信息学分析[J].北方园艺,2016,40(13):98.[doi:10.11937/bfyy.201613026]
FENG Lijuan,YIN Yanlei,YANG Xuemei,et al.Cloning and Bioinformatical Analysis of DAHPS Gene in Pomegranate Peel[J].Northern Horticulture,2016,40(21):98.[doi:10.11937/bfyy.201613026]
[10]张艳菊,刘奇,刘东,等.黄瓜棒孢叶斑病菌的毒力分化及AFLP分析[J].北方园艺,2016,40(16):92.[doi:10.11937/bfyy.201616025]
ZHANG Yanju,LIU Qi,LIU Dong,et al.Virulence Differentiation and AFLP Analysis of Corynespora cassiicola[J].Northern Horticulture,2016,40(21):92.[doi:10.11937/bfyy.201616025]
备注/Memo
作者简介：赵丽华（1972-），女，四川西昌人，博士，副教授，研究方向为园艺植物分子生物学及遗传育种。E-mail:1973zlh@163.com.