文献类型：期刊文献

中文题名：西伯利亚杏花芽酵母cDNA文库构建及质量评价

英文题名：Construction and quality evaluation of cDNA library of flower bud yeast from Prunus sibirica

作者：陈俊兴[1,2] 包文泉[2] 敖敦[2] 蔺悦[2] 陈一潇[2] 伊茹[2] 徐宛玉[3] 王淋[1]

第一作者：陈俊兴

机构：[1]中国林业科学研究院经济林研究所,河南郑州450003;[2]内蒙古农业大学林学院,内蒙古呼和浩特010000;[3]河南农业大学园艺学院,河南郑州450002

年份：2025

卷号：45

期号：2

起止页码：131-138

中文期刊名：中南林业科技大学学报

外文期刊名：Journal of Central South University of Forestry & Technology

收录：;北大核心:【北大核心2023】；

基金：国家自然科学基金项目(32360400);国家重点研发计划(2023YFD220030202,2018YFD1000606-3)。

语种：中文

中文关键词：西伯利亚杏;花芽;酵母杂交;cDNA文库

外文关键词：Prunus sibirica;flower buds;yeast hybrid;cDNA library

分类号：S727.32

摘要：【目的】西伯利亚杏Prunussibirica是我国重要的生态兼木本粮油树种,由于其花期早,易受春季“倒春寒”影响,导致其产量不稳,严重制约其产业的发展。本研究利用西伯利亚杏花芽为材料构建酵母杂交cDNA文库,该文库可用于西伯利亚杏花芽分化及休眠的转录因子和互作蛋白的筛选,为今后西伯利亚杏花芽分化及休眠分子调控机制研究提供技术支撑。【方法】以西伯利亚杏花芽分化及休眠过程中15个时期的花芽为材料,进行总RNA提取以及mRNA的分离与纯化,并利用SMART同源重组技术构建cDNA文库。将cDNA文库转化至7酵母感受态细胞中,即得到cDNA酵母文库。【结果】鉴定结果表明,cDNA文库库容为5.24×10^(7) CFU/mL,插入片段长度主要分布在750~2000 bp,且平均插入长度在1000 bp以上,重组率为95.83%,24个阳性克隆经测序比对后,成功检测到20个功能注释基因,其中有3个序列(钙结合蛋白CML31、组蛋白H3.3、转录调节因子ADA2)与植物休眠及低温胁迫相关。【结论】西伯利亚杏花芽分化及休眠过程的酵母杂交cDNA文库构建质量较高,具有完整的基因信息,且符合酵母文库的筛选标准。该文库的构建能够为解析西伯利亚杏花芽分化及休眠相关的分子调控网络机制奠定基础。
【Objective】Prunus sibirica is an important ecological and woody food and oil tree species in China.Due to its early flowering period,it is susceptible to the freezing damage of spring“inverted spring cold”,which results in unstable yield and seriously restricts the development of its industry.In this study,a yeast hybrid cDNA library was constructed from Prunus sibirica flower buds,which can be used for the molecular regulation of bud differentiation and dormancy and the screening of interacting proteins.The construction of this yeast library can provide technical support for future research on the molecular regulation of bud differentiation and dormancy in Prunus sibirica.【Method】In this study,total RNA was extracted and mRNA was isolated and purified from flower buds of Prunus sibirica during 15 periods of flower bud differentiation and dormancy,and cDNA libraries were constructed by using SMART homologous recombination technology.The cDNA library was transformed into yeast receptor cells to obtain the cDNA yeast library.【Result】The results of quality identification showed that the library capacity of the cDNA library was 5.24×10^(7)CFU/mL,the length of insert fragments was mainly distributed between 750-2000 bp,and the average insert length was above 1000 bp,the recombination rate was 95.83%,and 20 functionally annotated genes were successfully detected in the 24 positive clones by sequencing and comparison,and three of these Three sequences(calcium-binding protein CML31,histone H3.3,and transcriptional regulator ADA2)were related to plant dormancy and low-temperature stress.【Conclusion】The yeast hybrid cDNA library of Prunus sibirica flower buds differentiation and dormancy was constructed with high quality,complete gene information,and met the criteria for yeast library selection.The construction of this library can lay the foundation for analyzing the molecular regulatory network mechanism related to bud differentiation and dormancy in Prunus sibirica.