A number of studies show that environmental stress conditions increase abscisic acid (ABA) and hydrogen peroxide (H2O2) levels in plant cells. Despite this central role of ABA in altering stomatal aperture by regulati...A number of studies show that environmental stress conditions increase abscisic acid (ABA) and hydrogen peroxide (H2O2) levels in plant cells. Despite this central role of ABA in altering stomatal aperture by regulating guard cell ion transport, little is known concerning the relationship between ABA and H2O2 in signal transduction leading to stomatal movement. Epidermal strip bioassay illustrated that ABA- inhibited stomatal opening and ABA-induced stomatal closure were abolished partly by externally added catalase (CAT) or diphenylene iodonium (DPl), which are a H2O2 scavenger and a NADPH oxidase inhibitor respectively. In contrast, internally added CAT or DPI nearly completely or partly reversed ABA-induced closure in half-stoma. Consistent with these results, whole-cell patch-clamp analysis showed that intracellular application of CAT or DPI partly abolished ABA-inhibited inward K+ current across the plasma membrane of guard cells. H2O2 mimicked ABA to inhibit inward K+ current, an effect which was reversed by the addition of ascorbic acid (Vc) in patch clamping micropipettes. These results suggested that H2O2 mediated ABA-induced stomatal movement by targeting inward K+ channels at plasma membrane.展开更多
Epidermal bioassay demonstrated that benzylamine, a membrane-permeable weak base, can mimick hydrogen peroxide (H2O2) to induce stomatal closure, and butyric acid, a membrane-permeable weak acid, can partly abolish th...Epidermal bioassay demonstrated that benzylamine, a membrane-permeable weak base, can mimick hydrogen peroxide (H2O2) to induce stomatal closure, and butyric acid, a membrane-permeable weak acid, can partly abolish the H2O2-induced stomatal closure. Confocal pH mapping with the probe 5-(and-6)- carboxy seminaphthorhodafluor- 1 - acetoxymethylester (SNARF-1-AM) revealed that H2O2 leads to rapid changes in cytoplasmic and vacuolar pH in guard cells of Viola faba L, i. e. alkalinization of cytoplasmic areas occur red in parallel with a decrease of the vacuolar pH, and that butyric acid pretreatment can abolish alkalinization of cytoplasmic areas and acidification of vacuolar areas of guard cells challenged with H2O2. These results imply that the alkalinization of cytoplasm via efflux of cytosol protons into the vacuole in guard cells challenged with H2O2 is important at an early stage in the signal cascade leading to stomatal closure.展开更多
The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics anal...The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics analysis and t-test were used to perform statistical analysis on chloroplast number of different ploidy level in those stomatal guard cells mentioned above, and morphology identifying and chromosome counting were used to test accuracy of counting chloroplast number in stomatal guard cells. The chloroplast average number in stomatal guard cells was very similar among the different leaf positions on the same plant and among significantly among the different ploidy the different locations in the same stoma in the same variety. All the leaf, while the chloroplast number varied distributions of the chloroplast number in different ploidy stoma were normal distribution fitted. A correlation has been established between ploidy and chloroplast number in the stomatal guard cells. In every single stoma of microspore-derived plants, the chloroplast number for a haploid should not be more than 10, diploids 11 to 15, and polyploids more than 15. The accuracy of this method for identification of different ploidy plants was 93.93%. Furthermore, the accuracy of this method was reliable and did not vary with the plants growth conditions. Therefore, the chromosome ploidy of plants derived from microspore culture in cabbage, broccoli, and Chinese kale can be identified by simply counting the chloroplast number in stomatal guard cells.展开更多
In this study, the direct effects of exterior abscisic acid （ABA） on both calcium distribution of mesophyll cells and cytosolic calcium concentration of guard cells were examined. The distribution of Ca^2＋ localiza...In this study, the direct effects of exterior abscisic acid （ABA） on both calcium distribution of mesophyll cells and cytosolic calcium concentration of guard cells were examined. The distribution of Ca^2＋ localization were observed with calcium antimonate precipitate-electromicroscopic-cyto-chemical methods after treated with ABA and pretreated with ethylene glycol-bis-（2-aminoethylether）-N,N,N＇,N＇-tetraacetic acid （EGTA）, verapamil （Vp）, and trifluoperazine （TFP）. The laser scanning confocal microscopy was used to measure the cytosolic calcium concentrations of guard cells under different treatments. The results showed that the cytosolic Ca^2＋ concentration of mesophyll ceils was induced to increase by ABA, but to decrease in both outside cell and the vacuoles within 10 rain after treatments. The cytosolic calcium concentration of guard cells was increased gradually with the lag in treatment time. However, both EGTA and TFP could inverse those effects, indicating that the increase of cytosolic calcium induced by exterior ABA was mainly caused by calcium influx. The results also showed that calmodulin could influence both the calcium distribution of mesophyll cells and calcium concentration of guard cells. It shows that calmodulin participates in the process of ABA signal transduction, but the mechanism is not known as yet. The changes both calcium distribution of mesophyll cells and calcium concentration of guard cells further proved that the variations of cytosolic Ca^2＋ concentration induced by ABA were involved in the stomatal movements of maize seedlings.展开更多
Stomata play a crucial role in plants by controlling water status and responding to drought stress.However,simultaneously improving stomatal opening and drought tolerance has proven to be a significant challenge.To ad...Stomata play a crucial role in plants by controlling water status and responding to drought stress.However,simultaneously improving stomatal opening and drought tolerance has proven to be a significant challenge.To address this issue,we employed the OnGuard quantitative model,which accurately represents the mechanics and coordination of ion transporters in guard cells.With the guidance of OnGuard,we successfully engineered plants that overexpressed the main tonoplast Ca^(2+)-ATPase gene,ACA11,which promotes stomatal opening and enhances plant growth.Surprisingly,these transgenic plants also exhibited improved drought tolerance due to reduced water loss through their stomata.Again,OnGuard assisted us in understanding the mechanism behind the unexpected stomatal behaviors observed in the ACA11 overexpressing plants.Our study revealed that the overexpression of ACA11 facilitated the accumulation of Ca^(2+)in the vacuole,thereby influencing Ca^(2+)storage and leading to an enhanced Ca^(2+)elevation in response to abscisic acid.This regulatory cascade finely tunes stomatal responses,ultimately leading to enhanced drought tolerance.Our findings underscore the importance of tonoplast Ca^(2+)-ATPase in manipulating stomatal behavior and improving drought tolerance.Furthermore,these results highlight the diverse functions of tonoplast-localized ACA11 in response to different conditions,emphasizing its potential for future applications in plant enhancement.展开更多
基金National Natura1 Science Foundation of China (No. 39870372),StateKey Basic Research and Development Project (No.G1999011700)
文摘A number of studies show that environmental stress conditions increase abscisic acid (ABA) and hydrogen peroxide (H2O2) levels in plant cells. Despite this central role of ABA in altering stomatal aperture by regulating guard cell ion transport, little is known concerning the relationship between ABA and H2O2 in signal transduction leading to stomatal movement. Epidermal strip bioassay illustrated that ABA- inhibited stomatal opening and ABA-induced stomatal closure were abolished partly by externally added catalase (CAT) or diphenylene iodonium (DPl), which are a H2O2 scavenger and a NADPH oxidase inhibitor respectively. In contrast, internally added CAT or DPI nearly completely or partly reversed ABA-induced closure in half-stoma. Consistent with these results, whole-cell patch-clamp analysis showed that intracellular application of CAT or DPI partly abolished ABA-inhibited inward K+ current across the plasma membrane of guard cells. H2O2 mimicked ABA to inhibit inward K+ current, an effect which was reversed by the addition of ascorbic acid (Vc) in patch clamping micropipettes. These results suggested that H2O2 mediated ABA-induced stomatal movement by targeting inward K+ channels at plasma membrane.
文摘Epidermal bioassay demonstrated that benzylamine, a membrane-permeable weak base, can mimick hydrogen peroxide (H2O2) to induce stomatal closure, and butyric acid, a membrane-permeable weak acid, can partly abolish the H2O2-induced stomatal closure. Confocal pH mapping with the probe 5-(and-6)- carboxy seminaphthorhodafluor- 1 - acetoxymethylester (SNARF-1-AM) revealed that H2O2 leads to rapid changes in cytoplasmic and vacuolar pH in guard cells of Viola faba L, i. e. alkalinization of cytoplasmic areas occur red in parallel with a decrease of the vacuolar pH, and that butyric acid pretreatment can abolish alkalinization of cytoplasmic areas and acidification of vacuolar areas of guard cells challenged with H2O2. These results imply that the alkalinization of cytoplasm via efflux of cytosol protons into the vacuole in guard cells challenged with H2O2 is important at an early stage in the signal cascade leading to stomatal closure.
文摘The relationship between the ploidy level of microspore-derived plants and chloroplast number in stomatal guard cells was studied in cabbage, broccoli, and Chinese kale. In the experiment, distribution statistics analysis and t-test were used to perform statistical analysis on chloroplast number of different ploidy level in those stomatal guard cells mentioned above, and morphology identifying and chromosome counting were used to test accuracy of counting chloroplast number in stomatal guard cells. The chloroplast average number in stomatal guard cells was very similar among the different leaf positions on the same plant and among significantly among the different ploidy the different locations in the same stoma in the same variety. All the leaf, while the chloroplast number varied distributions of the chloroplast number in different ploidy stoma were normal distribution fitted. A correlation has been established between ploidy and chloroplast number in the stomatal guard cells. In every single stoma of microspore-derived plants, the chloroplast number for a haploid should not be more than 10, diploids 11 to 15, and polyploids more than 15. The accuracy of this method for identification of different ploidy plants was 93.93%. Furthermore, the accuracy of this method was reliable and did not vary with the plants growth conditions. Therefore, the chromosome ploidy of plants derived from microspore culture in cabbage, broccoli, and Chinese kale can be identified by simply counting the chloroplast number in stomatal guard cells.
基金supported by the Natural Science Foundation of Hebei Province,China(C2007000994)the National Key Technology R&D Program,China(2007BAD69B01).
文摘In this study, the direct effects of exterior abscisic acid （ABA） on both calcium distribution of mesophyll cells and cytosolic calcium concentration of guard cells were examined. The distribution of Ca^2＋ localization were observed with calcium antimonate precipitate-electromicroscopic-cyto-chemical methods after treated with ABA and pretreated with ethylene glycol-bis-（2-aminoethylether）-N,N,N＇,N＇-tetraacetic acid （EGTA）, verapamil （Vp）, and trifluoperazine （TFP）. The laser scanning confocal microscopy was used to measure the cytosolic calcium concentrations of guard cells under different treatments. The results showed that the cytosolic Ca^2＋ concentration of mesophyll ceils was induced to increase by ABA, but to decrease in both outside cell and the vacuoles within 10 rain after treatments. The cytosolic calcium concentration of guard cells was increased gradually with the lag in treatment time. However, both EGTA and TFP could inverse those effects, indicating that the increase of cytosolic calcium induced by exterior ABA was mainly caused by calcium influx. The results also showed that calmodulin could influence both the calcium distribution of mesophyll cells and calcium concentration of guard cells. It shows that calmodulin participates in the process of ABA signal transduction, but the mechanism is not known as yet. The changes both calcium distribution of mesophyll cells and calcium concentration of guard cells further proved that the variations of cytosolic Ca^2＋ concentration induced by ABA were involved in the stomatal movements of maize seedlings.
基金supported by the Zhejiang Provincial Natural Science Foundation(LR21C020001)the National Natural Science Foundation of China(32372017,31871537 and U2003115)+3 种基金the Hainan Seed Industry Laboratory(B21HJ0220)to Y.W.the Biotechnology and Biological Sciences Research Council(BBSRC)grants(BB/W001217/1)to M.R.B.and R.K.BBSRC grants(BB/S017348/1)Royal Society University Research Fellowship awards(URFR 211002)to R.K.
文摘Stomata play a crucial role in plants by controlling water status and responding to drought stress.However,simultaneously improving stomatal opening and drought tolerance has proven to be a significant challenge.To address this issue,we employed the OnGuard quantitative model,which accurately represents the mechanics and coordination of ion transporters in guard cells.With the guidance of OnGuard,we successfully engineered plants that overexpressed the main tonoplast Ca^(2+)-ATPase gene,ACA11,which promotes stomatal opening and enhances plant growth.Surprisingly,these transgenic plants also exhibited improved drought tolerance due to reduced water loss through their stomata.Again,OnGuard assisted us in understanding the mechanism behind the unexpected stomatal behaviors observed in the ACA11 overexpressing plants.Our study revealed that the overexpression of ACA11 facilitated the accumulation of Ca^(2+)in the vacuole,thereby influencing Ca^(2+)storage and leading to an enhanced Ca^(2+)elevation in response to abscisic acid.This regulatory cascade finely tunes stomatal responses,ultimately leading to enhanced drought tolerance.Our findings underscore the importance of tonoplast Ca^(2+)-ATPase in manipulating stomatal behavior and improving drought tolerance.Furthermore,these results highlight the diverse functions of tonoplast-localized ACA11 in response to different conditions,emphasizing its potential for future applications in plant enhancement.
