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林业科学 ›› 2006, Vol. 42 ›› Issue (1): 43-48.doi: 10.11707/j.1001-7488.20060108

• 论文及研究报告 • 上一篇    下一篇

油茶种子EST文库构建及主要表达基因的分析

谭晓风1,2 胡芳名1,2 谢禄山1 石明旺1,2 张党权1,2 乌云塔娜1,2   

  1. 1.中南林业科技大学经济林育种与栽培国家林业局重点实验室,长沙410004;2.中南林业科技大学资源与环境学院,长沙410004
  • 收稿日期:2004-12-09 修回日期:1900-01-01 出版日期:2006-01-25 发布日期:2006-01-25

Construction of EST Library and Analysis of Main Expressed Genes of Camellia oleifera Seeds

Tan Xiaofeng1,2,Hu Fangming1,2,Xie Lushan1,Shi Mingwang1,2,Zhang Dangquan1,2,Wuyun Tana1,2   

  1. 1.Key Lab of Non-Wood Forest Product of State Administration of Forestry Central-South Forestry University Changsha410004; 2. College of Resources and Environment, Central_South Forestry University Changsha410004
  • Received:2004-12-09 Revised:1900-01-01 Online:2006-01-25 Published:2006-01-25

摘要:

为研究油茶种子在油脂转化高峰期的基因表达并分离克隆与油脂合成等有关的重要基因,以油茶优良无性系湘林1号和湘林4号近成熟种子为材料构建cDNA文库;随机挑取2327个克隆进行3′端测序构建EST文库。将数据完整并无X、N的1979条cDNA序列与NCBI核酸数据库的非冗余序列进行同源性比较,确认763条cDNA序列与核酸数据库中其他物种的DNA序列具有很高或较高的同源性,可确认266种基因;有1216个克隆序列为未知功能的基因序列。各类基因表现出不同的表达丰度,其中贮藏蛋白基因、与基因表达调控相关的基因、抗逆相关基因、种子成熟和胚胎发育相关的基因等为高丰度表达,与脂肪酸代谢相关的基因等为中丰度表达,另外还有大量的其他基因和未知功能基因在种子中表达。各类基因表达的数量和趋势与种子接近发育成熟阶段相吻合。

关键词: 油茶, cDNA, EST, 文库, 表达基因

Abstract:

To investigate the gene expression and to separate clones which are important genes related to oil synthesis and so on during transforming peak of seed oil in Camellia oleifera, cDNA library was constructed using the nearly matured seeds of two clones, I.e. XL-1 and XL-4, and 2 327 clones were randomly selected for sequencing on the 3′-end to construct the EST library, and 1 979 sequences which are unabridged and without X or N were performed homology comparison with nucleic acid NR database. 763 cDNA sequences were highly homological to cDNA sequences of other plant species in database of NCBI, 266 kinds of genes have been identified and 1 216 clones of cDNA were unknown function of gene sequences. In the cDNA library of seeds of C. oleifera, different genes expressed different abundance. The genes related to storage protein, to expression and regulation, to resistance, to seed maturation and to embryo development were expressed in high abundance, while the genes related to metabolism of fatty acids etc. were expressed in medium abundance. The other genes responsible for many fragments and many genes unknown functions were expressed in different extent. The number and trends of expressed genes are suitable for the phase of C. oleifera seeds near to maturation.

Key words: Camellia oleifera, cDNA, EST, Library, Expressed genes