草莓DELLA蛋白的分析与克隆鉴定
毕业论文关键词：草莓；DELLA蛋白；克隆；基因表达
Analysis and cloning identification of strawberry DELLA protein
Abstract: Strawberry (Fragaria × ananassa Duch.)is a perennial Rosaceae herb plant whose fruit has high nutritional value. Gibberellins are often applied to regulate the growth of strawberry during production in order to improve the fruit yield and quality. DELLA protein is an important regulatory factor of gibberellin signal transduction. In the amino acid sequence of the DELLA protein. In this study, the conserved domain of DELLA protein was obtained from the database analysis, and it was used as the retrieval sequence for identifying the DELLA Protein Encoding Gene of forest Strawberry. The study finds that there are two DELLA protein-encoding genes in the forest strawberry genome, the phylogene tic tree shows that the strawberry DELLA protein has high homology with the homologous proteins in Arabidopsis thaliana and Solanum lycopersicum, and the expression analysis reveals that the expression trends of these two DELLA protein-encoding genes is similar in all tested tissues. At last, we extracted total RNA from the strawberries and reverse transcribed them to cDNA, then designed gene-specific primers and amplified two DELLA protein encoding genes of the strawberries, successfully constructed the strawberry DELLA protein gene overexpression vector, these results lay a solid foundation for the DELLA protein further functional research.
Key words: strawberry; DELLA protein; clone; gene expression
目 录
摘要1
关键词1
Abstract1
Key words1
引言1
1材料与方法3
1.1材料 3
1.2方法 3
1.2.1基因序列分析3
1.2.2基因表达分析3
1.2.3总RNA的提取和cDNA的合成3
1.2.4目的基因的克隆4
1.2.5构建基因表达载体4
2结果与分析4
2.1森林草莓DELLA 蛋白生物信息学分析5
2.1.1基因序列全长与同源性比较5
2.1.2基因芯片数据与基因表达热图 6
2.2森林草莓不同组织器官的Total RNA提取7
2.3草莓DELLA 蛋白编码基因克隆与过量表达载体构建 7
3讨论 8
致谢 9
参考文献10
图1草莓DELLA蛋白与拟南芥DELLA蛋白的序列比对6
图2不同植物来源DELLA蛋白的系统进化关系6
图3基因表达热图 7
图4 Plant Total RNA Isolation Kit Plus法提取的RNA电泳图7
图5草莓DELLA基因克隆与过量表达载体构建8
表1引物序列4
草莓DELLA蛋白的分析与克隆鉴定
植物从种子萌发到果实成熟的整个生命循环过程受到许多内源因子的调节。赤霉素(GA)作为一种重要的生长激素，能够调控植物胚根的生长、茎杆的伸长、叶片的延展、花的发育、植物次生代谢产物的合成、花器的分化和果实的成熟等各个方面[1]。在以上几个方向中，GA的最显著的调控方向是对植物株型的调控，外施GA或者植物体内GA浓度升高时可以使植物的茎干伸长，而当植物感受不到GA信号时，则产生矮化的表型。如在草莓的营养生长阶段，外施GA可以促进草莓匍匐茎的生长，而在着果阶段，施加GA可以促进花梗和叶柄的伸长，从而防止因植株生长过于紧簇而引起的病害。用GA处理后的草莓植株，其匍匐茎的数量显著提高，同时也使叶片数增加,而且使单个叶片的大小也明显增大。另外，GA对保护地草莓有打破休眠、促进植株发育、提早物候期和采收提前的作用，而且植株生长整齐，无矮化现象。
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