Functional Analysis Of CaHPR1 Gene Of Plant Defense Stresses Controlled By CaRop1

High temperature high humidity environment is associated with severe disease and insect disaster,plant in the long-term of the adaptive process,which is inevitable evolution out of the cooperative defense reaction mechanism.ROP is plant multi-function molecular switch,plays an important signal metabolic regulation function in plants.So far many reports that ROP have the functions in plant adversity stress and hormone response.Hydroxypyruvate reductase(HPR)is one of the key enzymes in plant light breathing which play important roles in the related process of light respiration and photosynthesis.There are reports that HPR is controlled by light,plant hormone,and plant senescence,and may be involved in the plant disease resistance defense reaction.This study focused on the synergy of pepper in high temperature tolerance and disease resistance.With the development of CaRop1 a lot of function and mechanism research on the basis study,through the analysis of CaHPR1 and CaRopl interaction,the sub-cellular localization of CaHPR1,the CaHPR1 organization specificity and preliminary study its functions in plant,which will connect CaHPR1,CaRop1 and plant heat stress tolerance and disease-resistant defense reaction,studying the functional of CaHPR1 gene in plant stresses response controlled by CaRop1.The main results as follows:1?Through verifying of CaHPR1 and CaRop1 interaction by reverse yeast two hybrid and plant in vivo fluorescence double molecular complementary experiment(BIFC),we found CaHPRl and DN-CaRop1 interaction,but no interaction with CA-CaRop1.2?Successfully constructed CaHPR1,DN-CaRop1 and CA-CaRop1 prokaryotic expression vectors and successfully induced expression,for the next step to carry out interaction in vitro test,enzyme activity determination and so on.3?Through the analysis of the CaHPR1 sequence and protein structure that it has D-isomer specific 2-hydroxyacid dehydrogenases NAD-binding domain,C terminal has SKL microbody alignment signal.4.Successfully constructed the vector of sub-cellular localization carrier pMDC83-CaHPR1,and through the gene gun bombardment transformation onion epidermal cells,fluorescence microscope observation found that its position in the cell membrane intima,a few scattered around in cell lining,further confirmed the possibility interaction of CaHPRl and CaRop1.5?Through the RT-PCR and RT-FQ-PCR method analysis CaHPR1 in plant tissue of the distribution situation,found that the main distribution in plant leaves,and in plant roots had not detected.This experiment results is the same as published reports,CaHPR1 as a key enzyme in the process of plant photorespiration,mainly exist in plant leaves and the cotyledons.6?Successfully constructed the gain-of-function overexpression vectors that carry CaHPRl gene and successful transformation of tobacco(K326).Preliminary analysis the functions of TO generation plants,found that CaHPR1 has certain effect in high temperature tolerance.Successfully constructed the loss of function VIGS vector pYL279-iCaHPR1,and successful transformation pepper seedlings,the transient expression results show that the CaHPRl function lack body TRV::CaHPRl is more sensitive in heat stress environment,further verify the CaHPRl relative to plant high temperature tolerance.In this study we verify the interaction of CaHPRl and DN-CaRopl,and through the tissue specificity analysis and protein sub-cellular localization,Preliminary understanding the CaHPRl basic information,and guide the development of the subsequent experiment.Through the preliminary function analysis,it is proved that CaHPR1 relative to plant high temperature tolerance.