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A Preliminary Study on Molecular Marker Detection and Proteomics of Space Mutation on Pepper

Author: ZhouChunYang
Tutor: LiJingFu
School: Northeast Agricultural University
Course: Olericulture
Keywords: Pepper Space mutation AFLP two-dimensional electrophoresis
CLC: S641.3
Type: Master's thesis
Year: 2012
Downloads: 54
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Pepper originated from tropical area of Latin America is an important vegetables, it is one kind of vegetables in five in China. Because the breeding of traditional pepper was limited in intraspecific, and the using genes increasingly concentrated on few germ plansm, with the constant renewal of varieties, genetic basis became more and more narrow. Though there were many bred variety, it would be limited in cultivate new varieties. Space mutation bredding was a new emerging breeding technology combined with space technology, biotechnology and agricultural breeding technology. It had a good extensive application prospect in mutation breeding of plants.Morphological study on space mutation pepper named one, two, three and their control were researched in this paper. We used molecular marker technology AFLP to detecte mutant molecular, and clone, sequence and analyze differential fragments of detected mutant. Maintime analysed pepper leaves total protein by two-dimensional electrophoresis. The results were as followed:1. Pepper variety of space mutation had some changes in agronomic characters compared with their controls. The plant height of space one and space three were higher than these in controls, the plant height of space two was lower than that in control; The shapes of leaves changed little; The leaf color of space one and two changed little, the leaf color of space three was darker than control; the fruit number per tree and fresh weight of three variety after mutation were more than controls, the diameter of root of three variety after mutation were thicker than controls.2. AFLP system were optimized which were suited for space mutation pepper, it obtained318differentially expressed genes according to the analysis of differential expresstion of100universal primers on three tested varieties genes using AFLP system. After recycling, cloning and sequencing, we got179specific expression genes, GSSs were used for sequencing analysis. Then we got68known function genes and used GO classification for sequencing analysis. We got specific GSSs after space mutation by means of the further analysis of known genes.3. by means of two-dimensional chromatography technology about protein, there were91protein spots after the contrast of space one and its control, among them, there were22significantly different spots,18expressions up-regulated,4expressions down-regulated; there were74protein spots after the contrast of space two and its control, among them, there were12significantly different spots,11expressions up-regulated,1expressions down-regulated; there were90protein spots after the contrast of space one and its control, among them, there were36significantly different spots,30expressions up-regulated,6expressions down-regulated. Mass spectrometry identification and database searching were carried out on39protein spots had significantly difference, then we obtained the sequence and concrete function using biological informatics means.

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