[1]赵建华,李浩霞,尹跃,等.枸杞酸性转化酶基因的克隆与表达[J].江苏农业学报,2015,(05):1140-1148.[doi:doi:10.3969/j.issn.1000-4440.2015.05.031]
　ZHAO Jian-hua,LI Hao-xia,YIN Yue,et al.Cloning and expression of acid invertase gene(LbAI) in wolfberry (Lycium barbarum L.)[J].,2015,(05):1140-1148.[doi:doi:10.3969/j.issn.1000-4440.2015.05.031]
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枸杞酸性转化酶基因的克隆与表达()
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江苏农业学报[ISSN:1006-6977/CN:61-1281/TN]

卷:
期数:
2015年05期
页码:
1140-1148
栏目:
园艺
出版日期:
2015-10-31

文章信息/Info

Title:
Cloning and expression of acid invertase gene(LbAI) in wolfberry (Lycium barbarum L.)
作者:
赵建华12; 李浩霞3; 尹跃2; 安巍2; 王华芳1; 王亚军2; 石志刚2
(1.北京林业大学生物科学与技术学院，北京100083；2.宁夏农林科学院/国家枸杞工程技术研究中心,宁夏银川750002；3.宁夏农林科学院荒漠化治理研究所，宁夏银川750002)
Author(s):
ZHAO Jian-hua12; LI Hao-xia3; YIN Yue2; AN Wei2; WANG Hua-fang1; WANG Ya-jun2; SHI Zhi-gang2
(1.College of Biological Sciences and Technology, Beijing Forestry University, Beijing100083, China;2.Ningxia Academy of Agricultural and Forestry Sciences/National Wolfberry Engineering Research Center, Yinchuan 750002, China;3.Desertification Control Research Institute, Ningxia Academy of Agricultural and Forestry Sciences, Yinchuan 750002, China)
关键词:
枸杞; 酸性转化酶; 基因克隆; 表达; 可溶性糖
Keywords:
wolfberry; acid invertase; gene cloning; expression; soluble sugar
分类号:
S567；Q785
DOI:
doi:10.3969/j.issn.1000-4440.2015.05.031
文献标志码:
A
摘要:
以枸杞果实为试验材料，利用RACE 技术克隆枸杞酸性转化酶基因LbAI的全长cDNA 序列，应用生物信息学软件分析LbAI预期编码蛋白质特征；采用气相色谱法和实时荧光定量PCR 技术，分析不同发育阶段枸杞果实及不同颜色成熟果实中可溶性糖含量及LbAI 在果实中的相对表达量。结果显示：LbAI 的cDNA 序列全长2 252 bp，开放阅读框（ORF）长1 920 bp，编码642 个氨基酸，LbAI编码的蛋白质相对分子量和等电点（pI）分别为70 600 和5.9，GenBank 登录号为KM191309。进化树分析结果显示，枸杞与马铃薯和番茄的亲缘关系最近，相似性在85%以上。随着果实生长发育，枸杞果实中果糖和葡萄糖含量不断升高，而蔗糖呈现出降低趋势；不同颜色枸杞果实中糖含量差异较大，成熟红色和黄色果实中果糖和葡萄糖含量显著高于黑色果实，但蔗糖含量在黑色果实中最高，在红色果实中最低；LbAI相对表达与果实中葡萄糖含量变化基本一致。表明LbAI基因在枸杞果实糖积累过程中具有重要的作用。
Abstract:
The acid invertase gene (LbAI) full-length cDNA sequence from the fruits of wolfberry (Lycium barbarum L.) was clone by rapid amplification of cDNA ends (RACE), the features of LbAI-encoded protein were analyzed by bioinformatics software, and the patterns of gene expression and accumulation of soluble sugars at different fruits growth stages and differently-colored fruits of wolfberry were analyzed by real-time fluorescent quantitative PCR (qRT-PCR) and gas chromatography（GC）. The full length of LbAI cDNA sequence was 2 252 bp, containing a 1920 bp open reading frame (ORF) which encoded 642 amino acids with a relative molecular mass of 70 600 and isoelectric point (pI) of 5.9. The phylogenetic tree showed that LbAI had the closest genetic relationship with Solanum tuberosum and S. lycopersicum，with the similarities above 85%. The contents of glucose and fructose increased along with the fruit growth and development, while sucrose content declined. The contents of glucose and fructose in red and yellow fruits were significantly higher than those in black fruit at maturity. The content of sucrose in black fruit was the highest and in red fruit was the lowest. The expression level of LbAI was consistent with the glucose content in fruits. It was suggested that LbAI might play an important role in sugar accumulation of wolfberry fruits.

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备注/Memo

备注/Memo:
收稿日期：2015-03-09 基金项目：国家自然科学基金项目(31360191、31060104)；宁夏自然科学基金项目(NZ13125)；宁夏农林科学院科技创新先导基金项目(NKYJ-14-07) 作者简介：赵建华（1977-），男，甘肃靖远人，博士，副研究员，主要从事枸杞种质创新与利用研究。（Tel）0951-6886792； (E-mail) zhaojianhua0943@163.com 通讯作者：王华芳，(E-mail) hfwang@bjfu.edu.cn
更新日期/Last Update: 2015-10-31