Construction and Identification of Flower Buds cDNA Library of Asian Lotus(Nelumbo nucifera)

doi:10.3969/j.issn.1009-7791.2016.01.010

Subtropical Plant Science ›› 2016, Vol. 45 ›› Issue (01): 48-52.DOI: 10.3969/j.issn.1009-7791.2016.01.010

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Construction and Identification of Flower Buds cDNA Library of Asian Lotus(Nelumbo nucifera)

WU Xuan,ZHANG Da-sheng,TIAN Dai-ke,ZHANG Wei-wei,LIU Qing-qing,LIU Feng-luan,CHEN Qing,ZHAO Xi-shuang,WU Shao-hua

(1.College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou 350002, Fujian China; 2.Shanghai Chenshan Plant Science Research Center, Shanghai Chenshan Botanical Garden, Chinese Academy of Sciences, Shanghai 201602, China; 3.Shanghai Key Laboratory for Plant Functional Genomics and Resources, Shanghai 201602, China; 4.Agriculture and Forestry Gardening, Shanghai Vocational College, Shanghai 201699, China)

Received:2016-01-13 Revised:2016-04-17 Online:2016-03-10 Published:2016-03-10
Contact: TIAN Dai-ke; WU Shao-hua

荷花花蕾cDNA文库构建及质量评价

吴璇,张大生,田代科,张微微,刘青青,刘凤栾,陈青,赵喜双,吴少华

(1.福建农林大学园艺学院，福建福州350002；2.中国科学院上海辰山植物科学研究中心，上海辰山植物园，上海 201602；3.上海市资源植物功能基因组学重点实验室，上海 201602；4.上海农林职业技术学院园艺园林系，上海 201699)

通讯作者: 田代科;吴少华
作者简介:吴璇，硕士研究生，从事观赏园艺研究；张大生，博士，副研究员，从事观赏园艺植物分子育种研究。
基金资助:
国家自然科学基金项目(31572158)；上海市绿化和市容管理局科技攻关项目(F112421, G152422)；上海市科学技术委员会课题(14DZ2260400)

Abstract

Abstract: To explore the transcriptome during floral organ development in Asian lotus(Nelumbo nucifera), the total RNA of flower buds was isolated from three typical lotus cultivars: N. nucifera ‘Honghu Honglian’(single flower), N. nucifera ‘Tangzhaotisi Lian’(double flower) and N. nucifera ‘Qianban Lian’(fully-doubled and thousand-petalled). The double-stranded cDNA was synthesized based on the extracted mRNA using SMART method, then digested with restriction enzyme SfiI, and finally purified and ligated to the pUC19 vector. Thereafter, the flower buds cDNA library was constructed and amplified. The constructed cDNA library had the titer of 1.12×106 pfu·mL-1. The insert sizes of the library were 500—2000 bp and the recombination efficiency of cDNA library was about 95%. The cDNA library would provide the fundamental information for studying on gene expressions and regulations during floral organ development in lotus flower buds.

Key words: lotus, flower buds, cDNA library

摘要： 为理解荷花Nelumbo nucifera花器官转录组表达情况，分别选取不同花型的代表品种‘洪湖红莲’Nelumbo nucifera ‘Honghu Honglian’(单瓣)、‘唐招提寺莲’N. nucifera ‘Tangzhaotisi Lian’(重瓣)和‘千瓣莲’N. nucifera ‘Qianban Lian’(千瓣及全重瓣)的花蕾为材料分离mRNA，利用SMART技术合成双链cDNA，经限制性内切酶SfiI酶切后回收去掉接头和500 bp以下片段的cDNA。将cDNA与pUC19载体连接，构建荷花花蕾cDNA文库。经检测，该文库容量为1.12 × 106 pfu·mL-1，插入片段大小集中在500～2000 bp，重组率为95%。该文库的成功构建为荷花花蕾期转录组数据的开发及其花器官发育相关基因的功能研究奠定了分子基础。

关键词: 荷花, 花蕾, cDNA文库

CLC Number:

Q943.2

WU Xuan,ZHANG Da-sheng,TIAN Dai-ke,ZHANG Wei-wei,LIU Qing-qing,LIU Feng-luan,CHEN Qing,ZHAO Xi-shuang,WU Shao-hua. Construction and Identification of Flower Buds cDNA Library of Asian Lotus(Nelumbo nucifera)[J]. Subtropical Plant Science, 2016, 45(01): 48-52.

吴璇,张大生,田代科,张微微,刘青青,刘凤栾,陈青,赵喜双,吴少华. 荷花花蕾cDNA文库构建及质量评价[J]. 亚热带植物科学, 2016, 45(01): 48-52.

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Construction and Identification of Flower Buds cDNA Library of Asian Lotus(Nelumbo nucifera)

荷花花蕾cDNA文库构建及质量评价

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