[1]王忠,杨清,汪仁,等.野生茄子cDNA-AFLP分析体系的优化和建立[J].北方园艺,2012,36(19):108-112.
　WANG Zhong,YANG Qing,WANG Ren,et al.Optimization and Establishment of cDNA-AFLP Technical System in Solanum melongena[J].Northern Horticulture,2012,36(19):108-112.
点击复制

野生茄子cDNA-AFLP分析体系的优化和建立

《北方园艺》[ISSN:1001-0009/CN:23-1247/S] 卷: 第36卷 期数: 2012年19期 页码: 108-112 栏目: 出版日期: 2012-10-30
Title:
Optimization and Establishment of cDNA-AFLP Technical System in Solanum melongena
文章编号:
1001-0009（2012）19-0108-05
作者:
王忠1; 2; 杨清2; 汪仁1; 郑玉红1
1.中国科学院 植物研究所，植物多样性与系统演化研究中心，江苏 南京210014；
2.南京农业大学 生命科学学院，作物遗传与种质创新国家重点实验室，江苏 南京 210095
Author(s):
WANG Zhong1; 2; YANG Qing2; WANG Ren1; ZHENG Yu-hong1
1.Research Center of Systematic and Evolutionary Botany，Institute of Botany，Chinese Academy of Sciences，Nanjing，Jiangsu 210014；
2.State Key Laboratory of Crop Genetics and Germplasm Enhancement，College of Life Sciences，Nanjing Agricultural University，Nanjing，Jiangsu 210095
关键词:
茄子; cDNA-AFLP; 体系优化
Keywords:
Solanum melongena; cDNA-AFLP; system optimization
分类号:
S 641.103.6
文献标志码:
A
摘要:
以高抗黄萎病的野生茄子托鲁巴姆的根为试材，对影响cDNA-AFLP分析体系的6个关键因素进行分析，以期建立适宜茄子的cDNA-AFLP分析体系，并得到清晰可辨的cDNA-AFLP图谱。结果表明：采用改良的Trizol法提取的RNA较完整，纯度较高；置换合成法合成双链cDNA成本低且效率高；连接产物取原液作为预扩增反应的模板，预扩增产物稀释50倍作为选择性扩增的模板，在25 μL反应体系中加入2 μL的MgCl2（25 mM）和0.2 μL的dNTPs（10 mM），可以获得带型丰富且重复性好的结果。
Abstract:
In this study，the key facters affecting cDNA-AFLP on establishing cDNA-AFLP analysis system for roots were analyzed in Solanum melongena with high resistance to Verticillium wilt，and a distinct electrophoretogram of cDNA-AFLP was obtained.The results showed that total RNA extracted by improved Trizol method was highly pure and complete.The double-stranded (ds) cDNA was synthesized using low-cost and efficient replacement and synthesis method.The abundant and repeatable cDNA-AFLP fingerpritings have been obtained when original ligation products were used as template in the preamplification PCR and the preamplification products were diluted to 50 times for selective amplification in 25 μL reaction volume containing 2 μL MgCl2 (25 mM) and 0.2 μL dNTPs (10 mM).

参考文献/References:

[1]Bachem C W B，Van Derhoeven R S，De Bruijn S M，et al.Visualization of differential gene expression using a novel method of RNA fingerprinting based on AFLP，analysis of gene expression during potato tuber development [J].Plant Journal，1996,9(5):745-753.

[2]Bachem C W B，Oomen R J F J，Visser R G F.Transcript imaging with cDNA-AFLP：a step-by-step protocol[J].Plant Molecular Biology Reporter,1998,16(2):157-173.

[3]Durrant W E，Rowland O，Piedras P，et al.cDNA-AFLP reveals a strikingoverlap in race-specific resistance and wound response gene expression profiles [J].The Plant Cell,2000,12(6):963-977.

[4]于虹,丁国华.cDNA-AFLP技术及在差异表达基因克隆上的应用[J].中国农学通报,2007,23(12):77-80.

[5]Nazeem P A，Jose Sherin，Sheeba N K，et al.Differential gene expression for bacterial wilt incidence in tomato (Solanum lycopersicum?L.)revealed by cDNA-AFLP analysis [J].Physiological and Molecular Plant Pathology,2011,8(3):1-7.

[6]Wang L，Zhou B R，Wu L L，et al.Differentially expressed genes in Populus simonii×Populus nigra in response to NaCl stress using cDNA-AFLP [J].Plant Science,2011,2(1):796-801.

[7]李怀芳,刘凤权,郭小密.园艺植物病理学[M].北京:中国农业大学出版社,2001.

[8]Halima B，Jacquemin J M，Baudoin J P，et al.Optimization of a reliable，fast，cheap and sensitive silver staining method to detect SSR markers in polyacrylamide gels[J].Biotechnology Agronomy Society and Environment,2006,10(2):77-81.

相似文献/References:

[1]赵静,夏海波,李艳青,等.根际促生菌YHN对番茄和茄子的促生作用研究[J].北方园艺,2014,38(04):30.
　ZHAO Jing,XIA Hai-bo,LI Yan-qing,et al.Study on Growth-promoting Effect of Plant Growth-promoting Rhizobacteria Strain YHN on?the Seedling of Tomato and Eggplant[J].Northern Horticulture,2014,38(19):30.
[2]王铁良,陈思,毛丽珍.渗灌条件下土壤水分对温室秋冬茬茄子生长效应[J].北方园艺,2014,38(04):45.
　WANG Tie-liang,CHEN Si,MAO Li-zhen.Effect of Soil Moisture on Growth of Greenhouse Eggplant With[J].Northern Horticulture,2014,38(19):45.
[3]樊绍翥,谷晓华,安凤霞.茄子耐热性状正态性遗传测验分析[J].北方园艺,2013,37(17):37.
　FAN Shao-zhu,GU Xiao-hua,AN Feng-xia.Normality Hereditary Testing Analysis of Thermal Tolerance Traits with Eggplant[J].Northern Horticulture,2013,37(19):37.
[4]李里,江涛,周安佩,等.滇杨侧芽休眠期基因表达多样性的cDNA-AFLP分析[J].北方园艺,2013,37(15):117.
　LI Li,JIANG Tao,ZHOU An-pei,et al.Diversity Analysis of Gene Expression at Dormancy Stage of Lateral Buds in Populus yunnanensis by cDNA-AFLP Markers[J].Northern Horticulture,2013,37(19):117.
[5]狄文伟,周 彤,赵 冰.不同引发剂对茄子种子引发效果的研究[J].北方园艺,2014,38(09):12.
　DI Wen-wei,ZHOU Tong,ZHAO Bing.Effects of Different Evocating Agent on the Evoked Response of Eggplant Seeds[J].Northern Horticulture,2014,38(19):12.
[6]叶景学,齐义杰,赵超男,等.等渗钠盐胁迫对茄子种子发芽特性的影响[J].北方园艺,2013,37(03):40.
　YE Jing-xue,QI Yi-jie,ZHAO Chao-nan,et al.Effect of the Iso-osmotic Sodium Salt Stress on Germinating Characteristic of the Eggplant Seeds[J].Northern Horticulture,2013,37(19):40.
[7]李烨,谢立波,姚建刚,等.沼渣与基质配比对茄子幼苗的影响[J].北方园艺,2012,36(03):28.
　LI Ye,XIE Li-bo,YAO Jian-gang,et al.Effects of Different Substrate with Biogas Residue Proportion on Seedlings of Eggplant[J].Northern Horticulture,2012,36(19):28.
[8]司鹏飞,詹德江,李广,等.设施条件下不同磷肥水平对茄子果实养分含量的影响[J].北方园艺,2012,36(03):151.
　SI Peng-fei,ZHAN De-jiang,LI Guang,et al.Effects of Different Phosphate Levels on Nutrient Content of Eggplant Fruits in Greenhouse[J].Northern Horticulture,2012,36(19):151.
[9]朱丽梅,崔群香,曹嘉懿,等.不同茄子品种对灰霉病的室内抗病性鉴定[J].北方园艺,2012,36(22):135.
　ZHU Li-mei,CUI Qun-xiang,CAO Jia-yi,et al.The Identification of Disease Resistance of Different Eggplant Varieties to Botrytis cinerea[J].Northern Horticulture,2012,36(19):135.
[10]王冰林,韩太利,李媛媛.根结线虫侵染对茄子苗期生理生化反应的影响[J].北方园艺,2012,36(23):139.
　WANG Bing-lin,HAN Tai-li,LI Yuan-yuan.Effects of Meloidogyne incognita Infection on Physiology and Biochemistry of Eggplant Seedling Leaves[J].Northern Horticulture,2012,36(19):139.

备注/Memo

第一作者简介：王忠（1979-），男，博士，助理研究员，研究方向为植物分子育种。E-mail:wangzhong@cnbg.net.

责任作者：杨清（1963-），男，博士，教授，博士生导师，研究方向植物基因组学与基因工程。E-mail:qyang19@njau.edu.cn.

更新日期/Last Update: 2014-09-09