|本期目录/Table of Contents|

[1]王兴红,高宝嘉,南宫自艳.松毛虫遗传多样性研究中AFLP反应体系的建立[J].应用与环境生物学报,2009,15(06):875-879.[doi:10.3724/SP.J.1145.2009.00875]
　WANG Xinghong,GAO Baojia,NANGONG Ziyan.Establishment of AFLP Fingerprinting in Genetic Diversity Analysis of Pine Caterpillars[J].Chinese Journal of Applied & Environmental Biology,2009,15(06):875-879.[doi:10.3724/SP.J.1145.2009.00875]
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松毛虫遗传多样性研究中AFLP反应体系的建立()
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《应用与环境生物学报》[ISSN:1006-687X/CN:51-1482/Q]

卷:
15卷
期数:
2009年06期
页码:
875-879
栏目:
技术与方法
出版日期:
2009-12-25

文章信息/Info

Title:
Establishment of AFLP Fingerprinting in Genetic Diversity Analysis of Pine Caterpillars
作者:
王兴红; 高宝嘉; 南宫自艳
（1河北农业大学林学院 保定 071000）
（2河北北方学院 张家口 075000）
Author(s):
WANG Xinghong;  GAO Baojia;  NANGONG Ziyan
(1College of Forestry, Agricultural University of Hebei, Baoding 071000, Hebei, China)
(2Hebei North University, Zhangjiakou 075000, Hebei, China)
关键词:
松毛虫; AFLP; 反应体系; 引物筛选; 遗传多样性
Keywords:
pine caterpillar;  AFLP;  reaction system;  primer screening;  genetic diversity
分类号:
S763.421 : Q3-3
DOI:
10.3724/SP.J.1145.2009.00875
文献标志码:
A
摘要:
以油松毛虫为材料，对T4 DNA连接酶不同用量、预扩增中的Mg2+浓度、dNTP浓度、引物浓度以及选择性扩增中的预扩增产物稀释倍数、Mg2+浓度、dNTP浓度、引物浓度和Taq酶浓度进行了比较分析. 最终建立了适合松毛虫的AFLP反应体系. 用优化的AFLP反应体系，以油松毛虫、赤松毛虫和马尾松毛虫为材料筛选引物，从81对引物组合中筛选出10对多态性高的引物组合. 图9 参17
Abstract:
In order to establish AFLP technique system by using the genomic DNA of Dendrolimus tabulaeformis Tsai et Liu as test material, different dosages of T4 DNA ligase, concentration of Mg2+, dNTP and primers were optimized in preamplification reaction system, and the dilute multiple of preamplification production and the concentration of Mg2+, dNTP, primers and different dosages of Taq DNA polymerase were also optimized in selective amplification reaction systerm. Very steady results were achieved. In the linking reaction, T4 DNA ligase 2 U was added to the ligation system. The preamplification system included Mg2+ 1.5 mmol/L, dNTP 0.2 mmol/L, PstI primer and TaqI primer 5.0×10-7 μmol/L, and 40 dilute multiple of preamplification production. The selective amplification systerm included Mg2+ 1.5 mmol/L, dNTP 0.2 mmol/L, PstI primer and TaqI primer 5.0×10-7 μmol/L, Taq DNA polymerase 1 U. By using D. tabulaeformis Tsai et Liu, D. spectabilis Butler and D. punctatus (Walker) as test materials, ten pairs of informative primers were identified from eighty-one pairs of primers by optimizing the AFLP reaction system. The result established a strong foundation for the AFLP analysis of pine caterpillar. Fig 9, Ref 17

参考文献/References:

1 Hou TQ (侯陶谦). Pine Caterpillars in China. Beijing, China (北京): Science Press, 1987. 29~78
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13 Kakouli-Duarte T, Casey DG, Burnell AM. Development of a diagnostic DNA probe for the fruit flies Ceratitis capitata and Ceratitis rosa (Diptera: Tephritidae) using AFLP. J Ecol Entomol, 2001, 94 (4): 989~997
14 Lu C (鲁成), Zhao AC (赵爱春), Xiang ZH (向仲怀), Wan CL (万春玲). AFLP analysis of genetic diversity of Bombyx mandarina in China. Zool Res (动物学研究), 2002, 23 (2): 166~172
15 SIMA YH (司马杨虎), Chen DX (陈大霞), Zhao AC (赵爱春), Lu C (鲁成), Xiang ZH (向仲怀). Study of segregation law of AFLP molecular marker in silkworm Bombyx mori. Sci Sericult (蚕业科学), 2003, 29 (2): 131~135
16 Zhang MZ (张民照), Kang L (康乐). Extraction of total DNA from locusts and optimization of reaction conditions for RAPD analysis. Zool Res (动物学研究), 2001, 22 (1): 20~26
17 Lu C (鲁成), Liu LB (刘连碧), Zhou ZY (周泽扬), Liao SY (廖顺尧), Feng LC (冯丽春), Xiang ZH (向仲怀). Polymorphism analysis of restricted fragment length of fibroin gene (PFB2.9) in silkworm Bombyx moril. J Southwest Agric Univ (西南农业大学学报), 1996, 18: 103~106

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备注/Memo

备注/Memo:
国家自然科学基金项目（No. 30771739）资助 Supported by the National Natural Science Foundation of China (No. 30771739)
更新日期/Last Update: 2009-12-24