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Study on Heterosis of Hybrid Brassica Campestris L.ssp.pekinensis Olsson Baiyin 2 with cDNA-AFLP

Author: LinJianLi
Tutor: ZhuZhengGe；GaoJianWei
School: Hebei Normal
Course: Genetics
Keywords: Chinese cabbage Heterosis cDNA-AFLP Differential gene expression
CLC: S634.1
Type: Master's thesis
Year: 2008
Downloads: 21
Quote: 0
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Abstract


Heterosis is a very important biological biosphere widespread phenomenon, and the successful use of in a variety of plants and animals. Brassica crops (Brassica) is the most common type of crop utilization of heterosis. Theoretically and experimentally investigate the domestic and foreign scholars have done a great deal in nearly a century, the study of the mechanism of heterosis, made some progress in the methods and guiding ideology, many doctrines. The mechanism of heterosis is still a the end of the solution of the mystery, there are many issues that need in-depth study. This article with 'white lead No. 2' hybrids and their parents differentially expressed genes cDNA-AFLP technique for materials applications, as follows: at the seedling stage, the rosette stage and cabbage period clipping leaves to extract leaf total RNA AFLP (amplified fragment length polymorphism) analysis, reverse transcribed into cDNA using reverse transcriptase. EcoR Ⅰ and Mse Ⅰ cDNA-AFLP operation of the double-stranded cDNA restriction enzyme, resulting sticky ends were connected on the artificial joint, pre-amplified and then with 64 pairs of selective primers amplified in 6% polyacrylamide gel stained by silver staining after gel electrophoresis fingerprint analysis. This study shows that there are significant gene expression differences between hybrids and parents. Hybrids and their parents are differentially expressed bands can be divided into four types: (1) the type of hybrid-specific expression; (2) the parents total silence type; (3) single expression of the same type (partial maternal and paternal partial) ; (4) single expression silent type (partial female parent and partial paternal). The results of this study: 29 differential fragments found in different developmental stages, five seedlings of differences fragments, differences in fragment 14 rosette stage, cabbage period 10 differential fragments. 19 expression in hybrids, three parents expressed 2 expression in one of the parents (female), five in one of the parents (paternal) expression. 29 sequenced fragments by homologous sequence for retrieval, and 13 can be found in the NCBI database homologous sequences, 16 is not in the database to find homologous sequences, presumably is a new gene. 13 known sequence analysis showed that these fragments involved in metabolism, signal transduction process. 29 of the protein sequence of the gene fragment by the homology alignment, and found the two gene fragments were to contain AP2/EREBP structural domain and PPR domains. And these two domains has an important role in the control of plant growth and development. In this paper, the cDNA-AFLP technique cabbage differentially expressed genes. Provide the basis of the results of this study will later further reveal the molecular mechanism of heterosis.

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