新型春性甘蓝型油菜开花相关基因的筛选及表达模式研究

发布时间：2018-05-11 23:04

本文选题：春性甘蓝型油菜 + 开花相关基因　；参考：《青海大学》2017年硕士论文

【摘要】：油菜是世界重要油料作物之一,也是我国重要的食用油来源,其在我国的种植面积和总产均居世界首位。而开花是影响油菜成熟期、产量及品质的重要因素。油菜开花相关基因的研究具有重要的理论价值和应用前景。本研究以特早熟春性甘蓝型油菜86号为研究材料,利用cDNA-AFLP技术筛选其苗期和蕾期差异表达的基因片段,克隆出差异表达基因的cDNA全长序列,并通过相对荧光定量PCR分析其在油菜不同时期不同组织的表达模式。主要研究结果如下:(1)cDNA-AFLP筛选分离出86号油菜苗期和蕾期差异表达的基因序列共64条,经BLAST比对,有54条序列均在油菜基因组序列中,其中的42条序列与拟南芥已知功能的基因序列相似度极高;另外12条序列虽在油菜基因组上,但其功能未知,也未在其它植物中分离出,是一些未知基因片段。(2)用RACE技术克隆出了未知基因片段中的84-23、85-25、128-1、107-23、171-1、174-1基因的3'端全长序列和128-1、107-23、167-12基因的5'端全长序列,并根据扩增出的5'和3'序列设计的全长引物克隆出了128-1和107-23的cDNA全长序列。其中克隆出的128-1基因cDNA全长为1071bp,序列分析发现,该基因与甘蓝型油菜中预测的基因XM_013850601.1只有1个碱基的同义突变。128-1基因编码349个氨基酸残基的蛋白,氨基酸同源比对发现该蛋白可能与TPR超家族蛋白合成有关。克隆出的107-23基因cDNA全长1345bp,与甘蓝型油菜中预测的XM_013752386.1基因编码区相似度很高,但在5'有1个碱基的突变,3'端有2个碱基的突变。107-23基因编码358个氨基酸残基的蛋白,氨基酸同源比对表明该蛋白可能是与开花相关的转录因子TCP2。(3)通过RT-qPCR分析差异表达基因128-1和107-23在86号油菜不同时期不同组织的表达情况,发现两基因在苗期、蕾期和花期各组织都有表达,其中128-1基因在花期花组织的表达量最高。本研究结果表明,新型春性甘蓝型油菜中筛选出的差异表达基因128-1和107-23可能与油菜开花基因有关,具体功能有待进一步转基因验证。
[Abstract]:Rape is one of the most important oil crops in the world, and it is also an important source of edible oil in China. Flowering is an important factor affecting the maturity, yield and quality of rape. The study of flowering related genes in rapeseed has important theoretical value and application prospect. In this study, early maturing Brassica napus 86 was used as the research material. CDNA-AFLP technique was used to screen the differentially expressed gene fragments at seedling and bud stages, and the full-length cDNA sequence of differentially expressed genes was cloned. The expression patterns in different tissues of rapeseed were analyzed by relative fluorescence quantitative PCR. The main results were as follows: a total of 64 differentially expressed genes were isolated from rape (Brassica napus L.) at seedling stage and bud stage by cDNA-AFLP screening. By BLAST alignment, 54 sequences were found in rapeseed genome sequence. 42 of these sequences were highly similar to those of Arabidopsis thaliana with known functions, and the other 12 sequences were unknown and not isolated from other plants, although they were in the genome of Brassica napus. RACE technique was used to clone the full length of 84-23128-1107-23171-1174-1 and 128-1107-23167-12, respectively. The full-length cDNA sequences of 128-1 and 107-23 were cloned according to the amplified 5 'and 3' full-length primers. The total length of 128-1 gene cDNA was 1071bp.The sequence analysis showed that this gene and the predicted gene XM_013850601.1 in Brassica napus had only one base of synonymous mutation .128-1 gene encoding 349 amino acid residues. Amino acid homology analysis revealed that this protein might be related to the synthesis of TPR superfamily proteins. The total length of cDNA of 107-23 gene was 1345bp. it was similar to the predicted coding region of XM_013752386.1 gene in Brassica napus, but there were two base mutations at the 3'end of 5'base mutation. 107-23 gene encoded 358 amino acid residues. Amino acid homology analysis indicated that the protein may be a transcription factor related to flowering, TCP2.3. the differential expression genes 128-1 and 107-23 were analyzed by RT-qPCR in different tissues of rape 86 at different stages, and the two genes were found to be in seedling stage. The expression of 128-1 gene was the highest in flower tissues. The results showed that the differentially expressed genes 128-1 and 107-23 may be related to the flowering genes of Brassica napus.
【学位授予单位】：青海大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：S565.4;Q943.2

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