The Study On Effects Of Methanol And Ethanol Stimulation On The Photosynthesis And Stomatal Of Faba Bean

Broad bean is an important crop in China, that’s very wide range of applications In food processing, feed production and other industries. Adaptability of Broad bean on Soil and climate patticularly extensive, It can be cultivated in red soil and yellow soil and neutral soil. Many studies have shown that application of methanol and ethanol to promote plant growth. The application of methanol growth promotion effect model plants such as tobacco and Arabidopsis and crop has been repeatedly confirmed. In Qi Chuanjiao’s studies shows methanol promotes of (?)ean growth and photosynthesis related genes transcription. Yunnan bean growing in the red soil cultivars as experimental materials in this study. Analysis of foliar methanol (CH3OH) and ethanol (CH3CH2OH) effects of faba bean growth and molecular mechanism of photosynthesis. The main results are as follows:(?)blade with3%(v/v) methanol and ethanol can promote growth of Faba Bean. Broad (?)plant height, fresh weight and dry weight and leaf number were significantly higher than (?)control. And ethanol on plant growth promoting effect than those of methanol. Foliar(?)and ethanol for3weeks, total soluble protein, soluble sugar, chlorophyll content were increased significantly. Leaf blade and leaf chlorophyll content of ethanol a, total soluble protein, total soluble sugar content was higher than that of methanol. After6hours of methanol and ethanol leaf photosynthetic indexes determination. Results showed that foliar methanol and ethanol can promote photosynthesis and leaf spraying leaves of Vicia faba, ethanol, photosynthetic rate, intercellular CO2concentration, stomatal conductance, transpiration rate were higher than those of methanol. Foliar methanol and ethanol and the content of H2O2and malondialdehyde (MDA) content in Vicia faba leaves decreased, protein carbonyl (PC) content increased, leaf and spraying alcohol Vicia faba leaves MDA and PC content was higher than that of methanol oxidation of ethanol, blade stress level is high in the spraying methanol. Analysis of the expression of3%methanol and ethanol within24hours of photosynthetic genes by RT-PCR, results show that methanol and ethanol on chlorophyll a/b binding protein (CAB), light accumulating protein complex (LHC), photosystem II and I related protein, plasmid vanillin (PC),(?)transfer protein Calvin cycle enzymes,(Rubisco, FBP) induced the transcription of different degree, that methanol and ethanol may stimulate photosynthesis by Vicia faba induced the expression of photosynthesis genes, so as to promote the growth and biomass accumulation of Vicia faba.To understand the foliar methanol and ethanol increased the molecular mechanism of stomatal conductance of leaves of Vicia faba. Results showed that foliar methanol and ethanol for0.5hours and stomatal conductance and transpiration rate decreased significantly opening. In2hours of recovery to the untreated control level, in6hours was significantly higher than that of control leaves and leaf spraying, ethanol is higher than that of methanol injection, then decreased to normal level. Phototropins (Vfphot),14-3-3protein (Vfl4-3-3) and plasma membrane H+-ATP enzyme (VHA) is a key part of regulation of stomatal opening signal pathway. RT-PCR analysis results show that ethanol on Vfphotla/lb,14-3-3a/c/d), the expression of VHA1/3induced. Methanol only on the expression of Vfphotla,14-3-3a/c/d, VHA2can be induced, immunoprecipitation (COIP) analysis showed that the phosphorylation level of VHA in leaves of methanol and ethanol increased significantly after6hours. The amount of14-3-3protein increased binding. Activity and hydrogen pump activity of VHA in leaves treated with methanol and ethanol for2hours after the increase, rose to the highest in6hours,12hours after the start to decline, in the whole period of24hours, the blade activity and hydrogen pump activity in leaf plasma membrane H+-ATP enzyme injection of ethanol is higher than that of the methanol injection. These results indicate that methanol and ethanol through interaction regulates the expression of Vicia faba leaf Vfphot,14-3-3protein and VHA and14-3-3protein and VHA to change the activity of VHA, thereby increasing the stomatal conductance and conductance.AMP and are inhibitors of plasma membrane H+-ATPase, combined with the former can inhibit14-3-3protein and phosphorylation of plasma membrane H+-ATPase and membrane H+-ATPase activity decreased. Phosphorylation of H2O2can inhibit plasma membrane H+-ATPase and its binding to14-3-3protein, thereby reducing the activity of H+-ATPase and reduce the in vitro expression of Vicia faba stomata opening. The results of the study showed the most significant effect of ethanol on Vicia faba leaf physiological and biochemical characteristics, in order to further investigate the plasma membrane ATP enzyme in the physiological characteristics of ethanol regulation of leaf stomata of function, containing VA, AMP and H2O2respectively in the3%ethanol solution of Vicia faba leaves of Vicia faba, found that6hours completely closed, stomatal conductance was significantly less than no processing control, showed that the addition of VA, AMP and H2O2completely inhibited ethanol increased stomatal opening. Analysis of H2O2content in leaves, results indicate the presence of adding VA, AMP and H2O2and the H2O2content of broad bean leaves was significantly higher than that of the untreated control and single leaf spraying ethanol. COIP analysis showed that H2O2was added in ethanol solution so that the level of interaction of14-3-3protein and VHA decreased, decreased activity and hydrogen pump activity of VHA, addition of VA and AMP decreased more significantly, these results indicated that ethanol through regulation of14-3-3protein and VHA interaction to change the activity of VHA and hydrogen pump, the regulation of stomatal opening and conductance.