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Study on Differential Proteome and XET Gene Associated with Petals Senescence in Dahlia Pinnata Cav

Author: ZuoXueQin
Tutor: TanJianZhong
School: Suzhou University
Course: Ornamental Plants and Horticulture
Keywords: Dahlia Senescence Differential proteome Protection of enzyme activity Clone
CLC: S682.11
Type: Master's thesis
Year: 2010
Downloads: 68
Quote: 2
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Abstract


Dahlia (Dahlia pinnata Cav) flowers, color and diverse and colorful highly ornamental garden plants cultivated with fresh cut flowers are widely used. But the Dahlia petals easy aging wilting, under natural conditions, a single flower shorter flowering stage, the best viewing period is only 4-5 days. Dahlia petal senescence mechanism and its regulation has always been the people's attention, and theoretically significant to improve the ornamental value of the adjustment of the Dahlia cut flowers fresh. In this paper, Dahlia the univalve type, yellow varieties as experimental materials, the establishment of a two-dimensional electrophoresis of proteins petals and differential proteomics technology system, Dahlia different the flowering period petals protein differences expression and protection of the enzyme activity period change, and differences in expression of proteins identified and cloned the full-length cDNA sequences of the senescence-associated protein Dahlia - xyloglucan within glycosyltransferase (XET) gene, made the following main experimental results: 1. on the basis of optimization of the Dahlia petals protein sample preparation techniques, application IPG-IEF/SDS-PAGE (2-DE) method, different periods of the Dahlia aging process petals protein separation, to obtain a clear, stable two-dimensional electrophoresis map, the ImageMaster 2D map analysis software dimensional electrophoresis profiles, 1240 protein spots were detected, of which the expression of more than 2-fold difference in 44; Second, differences in expression levels greater than 1.5 times protein spots in the overall downward trend in the aging process accounted for 52.4%, while 45.1% increase in the amount of aging late expression. 2 expression is selected from the 2-DE gels have a significant difference of 16 protein spots by gel digestion and mass spectrometry analysis and database searching, successfully identified one of the 11 different protein components. Existing research shows that these proteins are involved in the matter and energy metabolism, free radical scavenging and antioxidant effects, cytoskeleton formation and signal transduction physiological processes, may be related with the the Dahlia petals aging process regulation. According to differences in protein identification results using UV spectrophotometry, catalase (CAT) activity in the the Dahlia petals of the aging process in. At the same time, the tetrazolium (NBT) method and the guaiacol law, belong to the same system of protective enzymes superoxide dismutase (SOD) and peroxidase (POD) activity. The results show that: with the flower opening and senescence, CAT activity was increased and then decreased trend, this trend is consistent with dimensional electrophoresis profiles of CAT expression; SOD activity changes similar to CAT and POD activity showed early decline , the late rising trend. According to differences in protein identification results, the first time in the Dahlia petals aging process differentially expressed features found the Xyloglucan within glycosyltransferase (XET), the trend showed a gradual increase in the expression level, presumably The new functional proteins related to aging mechanisms. Application of bioinformatics and molecular biology experimental techniques to clone the the XET gene Dahlia full-length cDNA sequence, and the sequence size of 882 bp, encoding 293 amino acids; clustering analysis showed that the cDNA sequence of the cloned Dahlia XET The tested 12 dicotyledons homology is higher, and highly conserved amino acid sequence of the XET protein.

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Perennial Flowers > Perennial Flowers Class > Chrysanthemum
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