贵州大学生命科学学院/农业生物工程研究院/山地植物资源保护与种质创新教育部重点实验室/ 贵州省农业生物工程重点实验室，贵阳550025
马思宇，E-mail：CMMC23@163.com
宋莉，E-mail：lpsssl@126.com
S541+.6
国家自然科学基金项目（32260338；31660685）
The Key laboratory of Plant Resource Conservation and Germplasm Innovation in Mountainous Region(Ministry of Education)/Institute of Agro-bioengineering/College of Life Sciences， Guizhou University， Guiyang 550025， China
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为研究百脉根（Lotus corniculatus L.）C2钙依赖蛋白激酶基因LcC2DP1在不定根形成过程中的功能，通过RACE法从百脉根中克隆LcC2DP1基因，利用qRT-PCR检测其时空表达模式，并通过农杆菌介导的瞬时表达系统在百脉根中过量表达LcC2DP1并鉴定其功能。结果显示：LcC2DP1基因全长705 bp，编码235个氨基酸，分子质量为25.95 ku，与蒺藜苜蓿同源性最高（82%）；在百脉根不定根分化过程中持续表达，表达部位为根、茎和叶片；与野生型亲本（WT）相比，转LcC2DP1基因百脉根（TP）的不定根分化提前1~2 d；在不定根分化的9~15 d ，其总根长分别是WT的168%、155%，根体积分别是WT的249%、161%，根尖数分别是WT的156%、137%。TP百脉根的总根长（P<0.01）、根体积（P<0.01）和根尖数（P<0.05）表现出一定的发育优势，表明LcC2DP1基因可能与百脉根不定根发育调控相关。
The LcC2DP1 gene was cloned from the Lotus corniculatus L.by RACE method to study the function of C2 calcium dependent protein kinase gene LcC2DP1 of Lotus corniculatus L.in the process of adventitious root development.The spatial expression mode of LcC2DP1 gene was detected with qRT-PCR.The overexpression and function of LcC2DP were identified by Agrobacterium mediated transient expression system.The results showed that the full length of LcC2DP1 gene was 705 bp，encoding 235 amino acids.Its molecular weight was 25.95 ku，with the highest homology of Medicago truncatula （82%）.It was continuously expressed during the differentiation of adventitious roots in Lotus corniculatus L..The expression sites were roots，stems and leaves.Compared with wild-type parents （WT），the adventitious root differentiation of the LcC2DP1 transgenic Lotus corniculatus L. plants（TP） was 1-2 days earlier.On the 9th to 15th day of adventitious root differentiation，the total root length was 168% and 155% of WT，the root volume was 249% and 161% of WT，and the number of root tips was 156% and 137% of WT，respectively.The total root length （P<0.01），root volume （P<0.01） and number of root tips （P<0.05） of TP root had certain advantages of development，indicating that LcC2DP1 gene may be related to the regulation of adventitious root development in Lotus corniculatus L..
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马思宇,肖芳斌,罗雪,韦飘,宋莉.百脉根不定根发育相关基因LcC2DP1的克隆与功能初步分析[J].华中农业大学学报,2023,42(1):42-50
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- 收稿日期:2022-04-07
- 在线发布日期: 2023-02-22
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