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Physiological and Biochemical and Proteomics Preliminary Analysis of Ipomoea Cairica (Linn.) Sweet under Different Allelochemicals Stresses

Author: SunYanJie
Tutor: ZhuangDongHong;MaRuiJun
School: Shantou University
Course: Biochemistry and Molecular Biology
Keywords: Cairica Allelopathy Protective enzyme MDA Two-dimensional electrophoresis
CLC: S451
Type: Master's thesis
Year: 2011
Downloads: 83
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Abstract


In this paper, four kinds of monomers allelochemicals were theophylline, vanillic acid, (Z) -2 - (2,3 - dimethoxyphenyl methylene) -6 - hydroxybenzofuran -3 (2H ) - one (No. 081110) 'and (Z) -2 - (3 - pyridyl-methylene) -4,6 - dimethoxy-benzofuran -3 (2H) - one (No. 090 710) Processing Guangdong and the most serious hazards throughout southern China, one of the invasive weed cairica (Ipomoea cairica (Linn.) Sweet), to protect the enzyme system change is one of the indicators were extracted simultaneously by four kinds of allelochemicals 1 day after treatment The leaves and apical cairica total protein, two-dimensional electrophoresis, protein spots in order to change to another index, to observe these four kinds of allelochemicals whether it can effectively influence cairica these two indicators, while observing the different allelochemicals on root tips and leaves cairica proteins affect whether a certain regularity, to screen out cairica with specificity or efficiency of allelopathic substances, so our findings from a more practical application Further, for the development of botanical herbicides basis. In this study, the role of different allelochemicals under cairica chlorophyll content, superoxide dismutase (SOD), catalase (CAT), peroxidase (POD), and malondialdehyde (MDA) were determined, and further use of two-dimensional electrophoresis technique cairica allelochemicals in four kinds of proteins under the effect of changes in research, testing results show that in setting the concentration of allelochemicals 2mmol · L-1, the cairica chlorophyll content did not change significantly in the early stage of incubation, as prolonged incubation time compared with the control treatment groups were significantly yellowing leaves; including processed by 081,110 cairica leaf edge in the first 7 days of culture dark spots . By measuring the treatment group cairica protective enzyme showed that in addition to 090,710, the other three kinds of allelochemicals on cairica have a certain impact, expressed as SOD, CAT are first increased and then decreased; POD activity in addition to vanilla first decreased and then increased acid outside its Yudu Huang is now increased and then decreased; MDA show a continued decline in addition to 090,710, the remaining three kinds of allelochemicals showed a rising trend. Cairica roots and leaves for protein extraction method, two-dimensional electrophoresis conditions were optimized. The results showed that: Weigh 0.08g of fresh root tip, adding 1.6ml extract [10% trichloroacetic acid (TCA) was dissolved in acetone and containing 0.07% of β-mercaptoethanol] Isoelectric focusing [pH (3-10) gradient strips (11cm) Two-dimensional electrophoresis followed by silver staining] appropriate protein concentration of the sample 40μg; isoelectric focusing conditions 30000V · h when ,2-DE protein maps and clear points. And four kinds of allelochemicals on cairica leaves and apical protein group has a significant impact, including the use of acid on cairica vanilla leaf processing, can be obtained by visual observation of significant changes in 31 spots, which New spots one, raised spots 21, down nine spots; apical process significantly changes available 22 spots, including new spots 2, missing spots one, raised spots 18, down one spot . The use of theophylline on cairica leaves are processed by visual observation obtained 27 distinct change spots, including new spots 5, up 19 spots, down 3 spots; apical treatment available 16 distinct spots, including new spots 12, down four spots. Using (Z) -2 - (2,3 - dimethoxyphenyl methylene)-6 - hydroxy-benzofuran -3 (2H) - one (No. 081110) leaves the cairica processed through the eye observe a significant change available 29 spots, including missing spots 7, raised spots 4, down 18 spots; apical treatment available 31 distinct spots, including new spots 5, raised spots 10, down spots 16. Using (Z) -2 - (3 - pyridyl-methylene) -4,6 - dimethoxy-benzofuran -3 (2H) - one (No. 090 710) for processing the leaves cairica, by visual observation available 33 spots changed significantly, which has eight missing spots, raised spots 6, down 19 spots; apical significant change in processing available 30 spots, including new spots 9, up 18 spots months, down three spots. Compare different allelochemicals on cairica apical and leaves wondering if the protein has a certain regularity, found that treatment cairica by 090,710, mainly affect the signal rotation system; through acid treatment theophylline and vanilla cairica, mainly affecting the energy system and antioxidant systems; cairica processed by 081,110 of its regularity is unknown, needs further study. Wherein the functions of these proteins involved in photosynthesis, antioxidant, energy metabolism, signal transduction and gene transcription. Chief among these proteins are: cytochrome synthase, citric acid dehydrogenase, ATP synthase subunit intron encoded egg factor, glutamate receptor, ribosomal protein S11, these proteins and plant adaptation to environmental stress Each relationship. The results showed that: 1.4 kinds of allelochemicals, in addition to 090,710 pairs cairica protective enzyme system is not sensitive, the other three kinds of allelochemicals have a certain influence. 2 total protein extraction method optimization results as: TCA-acetone extraction cairica Root and leaf total protein is the best. 3-dimensional electrophoresis Optimization Results: fresh tissue 0.08g, adding 1.6ml extract, isoelectric focusing pH (3-10) gradient strips (11cm) two-dimensional electrophoresis followed by silver staining of the appropriate protein concentration of the sample 40μg ; isoelectric focusing conditions 30000V · h when ,2-DE protein maps and clear points. 4.4 kinds of allelochemicals are all cairica apical and leaf protein has an impact, which after acid treatment theophylline and vanilla cairica proteins, mainly affecting the energy system and antioxidant systems; treated by 090,710 five claw dragon, mainly affecting the signal rotation system; cairica processed by 081,110 of its ominous regularity, needs further study. This indicates that they are likely to become a more suitable alternative herbicide plant origin allelopathic substance.

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CLC: > Agricultural Sciences > Plant Protection > Harmful plants and their removal > Weeds
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