| It was carried out with a type of exotic invasive plant Ipomoea cairica (Linn.) Sweet which did great damage to the nature in the east of Guangdong and North China. The monomer allelochemicals used were vanillic acid, theophylline, (Z)-2-(2,3-dimethoxyphenyl methylene)-6-hydroxyl group benzofuran-3-(2H)-ketone(No.081110) and (Z)-2-(3-pyridylmethylene)-4, 6-dimethoxybenzofuran-3-(2H)-ketone (No.090710). To study the effects of allelochemicals on the receptor I. cairica and test the most effective allelochemical, there took two steps to test the change of protection enzymes and spots of protein. The total protein content was extacted from leaves and roots of I. cairica treated for 1 day with the four monomer allelochemicals and two dimensional electrophoresis. The aim of this study was to find special and effective allelochemicals and lay the groundwork for the development of botanical herbicide.The contents of chlorophyll, Superoxide Dismutase (SOD), Catalase (CAT), Peroxidase (POD), Malondialdehyde(MDA) in I.cairica under the influence of different allelochemicals were determined. The changes of proteins in I.cairica under the influence of different allelochemicals were researched using two dimensional electrophoresis. The results indicated:(1) When the concentrations of all the allelochemicals were 2mmol·L-1, the contents of chlorophyll had no significant changes in the preliminary stage. By contrasting to the control groups, the leaves turned yellow obviously in the study groups as time prolonging. And the margin of leaves developed black speckle on the 7th day with the treatment of 081110. (2) Except NO.090710, the rest allelochemicals had certain effects on I.cairica. The activities of SOD and CAT increased at first and then decreased. (3) POD activity with vanillic acid treatment decreased at first and then increased, and the trend was contrary to the remaining allelochemicals. (4) The content of MDA in No.090710 decreased steadily, they kept going increased with the rest allelochemicals treatment.In addition, the ways how to extract protein from leaves and roots of I.cairica and the optimization of two dimensional electrophoresis condition were researched. At first, added 1.6ml extract (acetone solution,10% trichloroacetic acid,0.07%β-mercaptoethanol) into 0.08g fresh root tip tissues. The proper protein sample concentration was 40ug with the isoelectrofocusing on 11cm gels with a pH3-10 gradient and electrophoresis silver staining. Many and clear protein spots of 2-DE map were observed with 30000V·h isoelectrofocusing. The four allelochemicals had significant effects on proteomics in leaves and root of I.cairica. With the treatments of vanillic acid, theophylline,081110 and 090710 on leaves, obviously changed protein spots were observed with the unaided eye. The number of up-regulated protein spot was 21,19,4 and 6, the number of down-regulated protein spot 9,3,18 and 19. Only the first two allelochemicals developed 1 and 5 new protein spots respectively, and the last two allelochemicals developed 7 and 8 disappeared protein spots respectively. With the treatments of vanillic acid, theophylline, 081110 and 090710 on root tip tissues, obviously changed protein spots were also observed with the unaided eye. The number of up-regulated protein spot was 18,0,10 and 18, the number of down-regulated protein spot 1,4,16 and 3, new protein spot 2,12,5 and 9. Only the vanillic acid treament developed 1 disappeared protein spot. The roles of these proteins involved photosynthesis, antioxidation, energy transformation, signal transduction and gene transcription. The main proteins included cytochrome synthase, insocitratedehydrogenase, ATP synthase subunit, intron coding protein, glutamate receptor and ribosomal protein S11. The results suggested that these proteins related to the adaptive capacity of plant to environmental stress.So the study got the follow results:1. Except 090710, the rest allelochemicals had certain effects on antioxidant system of I.cairica.2. The best way to extract proteins from leaves and roots of I.cairica:TCA-acetone solution.3. The optimization of two dimensional electrophoresis condition:added 1.6ml extract into 0.08g fresh root tip tissues. The proper protein sample concentration was 40ug with the isoelectrofocusing on 11cm gels with a pH3-10 gradient and two dimensional electrophoresis silver staining, Many and clear protein spots of 2-DE map were observed with 30000V·h isoelectrofocusing.4. The four types of allelochemicals had effects on proteins in leaves and roots of I.cairica. The protein of I.cairica are treated with theophylline and vanilla acid mainly affect the energy system and antioxidant system; The protein of I.cairica is treated with 090710 mainly affect the signal recognition and rotation system; The protein of I.cairica is treated with 081110 mainly affect don't clear, we need study in future. That means they all can become allelochemicals to make botanical herbicides. |
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