| PEPCase is an key enzyme fixing CO2 in the C4 photosynthetic pathway, which has an important effect on the high photosynthetic efficiency of C4 crops. High expression of transgenic maize-C4-phosphoenolpyruvate carboxylase gene rice (high-expressing transgenic corn C4 pepc rice) are better than no transgenic rice original seed in physiological traits, photosynthetic indexes and yield traits. In order to research the physiological mechanism of C4 pepc in C3 plant rice, this article use high expression corn C4 pepc rice (PC) and untransformed rice plant Kitaake (WT) as material, cell suspension systems was established by adding the effects chemical reagents such as phospholipid enzyme C(PLC) inhibitor sulfate new mold pigment (Neomycin sulfate, NS), phospholipid enzyme D (PLD) inhibitor butanol (1-butanol), calcium and the calcium of a flat iron plate for making cakes mixture glycol double (2-amino ether) four acetate ethylene glycol-bis (2-aminoethylether)-N,N,N',N'-tetraacetic acid, EGTA) and the effects were studies on cell within PEPC enzyme activity, related signal molecular NO, hydrogen peroxide(H2O2) and the calcium of effect in the cell level. The results were found as following:1.Establishment of cell suspension system PC and WT:2mg/L 2,4-D and 1mg/L 6-BA were added to induce the callus from the rice seed, then put the callus into the mediums of regeneration with hormones such as 2-0.5mg/L 2,4-D,2-0.2 mg/L 6-BA,1 mg/L ABA), however reduce outside source hormone of application step by step, finally the shaped,dispersed and particle callus can get to transfer to liquid medium adding 0.5mg/L 2,4-D,0.2 mg/L 6-BA,1 mg/L ABA at 28℃. The cell suspension system for rice material can be established stably after 6 weeks by adding generation new liquid of proportion for 3.1orriginally.2.Adding neomycin alone or n-butyl alcohol for one hour to inhibit PA in the cell, PEPC enzyme activity and NO, H2O2 concentration in PC were reduced depend on the concentration and time. After adding Mas for 1h to increase PA in the cell, the enzyme activity and NO, H2O2 in the cell of PC were reduced, so did the glucose effects in PC.3.In the cell, after adding both the n-butanol and neomycin in PC for 1h, the PEPC activities of PC were increased, while the NO, H2O2 content were decreased; When adding the Mas in PC, PEPC enzyme activity, NO, H2O2 concentration of PC were increased; the effects of glucose was similarly to that of Mas. At the same time, it appeared more Ca2+ concentration in the cell sinigicantly.4. When the 1-butanol, neomycin and EGTA were added at the same time in PC, PEPCase activities NO and H2O2 contents of PC were increased. Although the calcium ion releasing after mentioned above inhibitors in PC were reduced much lower than those only adding EGTA, the Ca2+ concentration in PC was still slightly increased as compared to PC-control without adding EGTA.The results showed that endogenous concentrations of calcium in the cell of PC was more related to regulate of PEPC activities due to the adding concentration of EGTA in this paper can partly inhibit the bursting of calcium sink. |
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