Tyrosine decarboxylase(TyDC)converts tyrosine to tyramine and plays a crucial role in secondary metabolite reactions,development,and stress responses in plants.Currently,the biological role of TyDC proteins from trees...Tyrosine decarboxylase(TyDC)converts tyrosine to tyramine and plays a crucial role in secondary metabolite reactions,development,and stress responses in plants.Currently,the biological role of TyDC proteins from trees is unknown.This study provided evidence showing that poplar PaTyDC4 functions in wood development and drought stress response.PaTyDC4 is preferentially expressed in wood-forming cells in stems.Overexpression of PaTyDC4 in poplars under the control of a xylem-specific promoter resulted in an increase in the ratio of xylem to phloem width,vessel cell area,and lignin accumulation in the stems.Biochemical assays revealed that PaTyDC4 was a component of the PaC3H17-PaMYB199 module-mediated pathway.In poplar stems,Pa TyDC4 expression was directly suppressed by PaMYB199,which was attenuated by the interaction between PaC3H17 and PaMYB199.In addition,Pa TyDC4 overexpression lines showed stronger drought tolerance than the wild-type lines,with higher photosynthetic capacity and lower levels of H_(2)O_(2).These results indicate that PaTyDC4 promotes xylem differentiation and lignin deposition during secondary growth and confers drought tolerance.Our findings may be useful for the genetic modification of biomass and drought resistance in trees.展开更多
While biochar amendment enhances plant productivity and water-use efficiency(WUE),particularly under waterlimited conditions,the specific mechanisms driving these benefits remain unclear.Thus,the present study aims to...While biochar amendment enhances plant productivity and water-use efficiency(WUE),particularly under waterlimited conditions,the specific mechanisms driving these benefits remain unclear.Thus,the present study aims to elucidate the synergistic effects of biochar and reduced irrigation on maize(Zea mays L.)plants,focusing on xylem composition,root-to-shoot signaling,stomatal behavior,and WUE.Maize plants were cultivated in splitroot pots filled with clay loam soil,amended by either wheat-straw biochar(WSB)or softwood biochar(SWB)at 2%(w/w).Plants received full irrigation(FI),deficit irrigation(DI),or partial root-zone drying rrigation(PRD)from the 4-leaf to the grain-filling stage.Our results revealed that the WSB amendment significantly enhanced plant water status,biomass accumulation,and WUE under reduced irrigation,particularly when combined with PRD.Although reduced irrigation inhibited photosynthesis,it enhanced WUE by modulating stomatal morphology and conductance.Biochar amendment combined with reduced rrigation significantly increased xylem K^(+),Ca^(2+),Mg^(2+),NO_(3)^(-),Cl^(-),PO_(4)^(3-),and SO_(4)^(2-)-but decreased Na+,which in turn lowered xylem pH.Moreover,biochar amendment and especially WSB amendment further increased abscisic acid(ABA)contents in both leaf and xylem sap under reduced irrigation conditions due to changes in xylem ionic constituents and pH.The synergistic interactions between xylem components and ABA led to refined adjustments in stomatal size and density,thereby affecting stomatal conductance and ultimately improving the WUE of maize plants at different scales.The combined application of WSB and PRD can,therefore,emerge as a promising approach for improving the overall plant performance of maize plants with increased stomatal adaptations and WUE,especially under water-limited conditions.展开更多
Lipids may play an important role in preventing gas embolisms by coating nanobubbles in xylem sap.Few studies on xylem sap lipids have been reported for temperate plants,and it remain unclear whether sap lipids have a...Lipids may play an important role in preventing gas embolisms by coating nanobubbles in xylem sap.Few studies on xylem sap lipids have been reported for temperate plants,and it remain unclear whether sap lipids have adaptational significance in tropical plants.In this study,we quantify the lipid composition of xylem sap for angiosperm species from a tropical savanna(seven species)and a seasonal rainforest(five species)using mass spectrometry.We found that all twelve species studied contained lipids in their xylem sap,including galactolipids,phospholipids and triacylglycerol,with a total lipid concentration ranging from 0.09 to 0.26 nmol/L.There was no difference in lipid concentration or composition between plants from the two sites,and the lipid concentration was negatively related to species’open vessel volume.Furthermore,savanna species showed little variation in lipid composition between the dry and the rainy season.These results support the hypothesis that xylem sap lipids are derived from the cytoplasm of individual conduit cells,remain trapped inside individual conduits,and undergo few changes in composition over consecutive seasons.A xylem sap lipidomic data set,which includes 12 tropical tree species from this study and 11 temperate tree species from literature,revealed no phylogenetic signals in lipid composition for these species.This study fills a knowledge gap in the lipid content of xylem sap in tropical trees and provides additional support for their common distribution in xylem sap of woody angiosperms.It appears that xylem sap lipids have no adaptive significance.展开更多
The xylem undergoes physiological changes in response to various environmental conditions during the process of plant growth.To understand these physiological changes,it is extremely important to observe the transport...The xylem undergoes physiological changes in response to various environmental conditions during the process of plant growth.To understand these physiological changes,it is extremely important to observe the transport of xylem.In this study,the distribution and structure of vascular bundle in Lilium lancifolium were observed using the method of semithin section.Methods for introducing a fluorescent tracer into the xylem of the stems were evaluated.Then,the transport rule of 5(6)-Carboxyfluorescein diacetate(CFDA)in the xylem of the stem of L.lancifolium was studied by fluorescence dye in live cells tracer technology.The results showed that the vascular bundles of L.lancifolium were scattered in the basic tissue,the peripheral vascular bundles were smaller and densely distributed,and the closer to the center,the larger the volume of vascular bundles and the more sparsely distributed.The vascular bundles of L.lancifolium are limited external tenacity vascular bundles,which are composed of phloem and xylem.The most suitable method for CFDA labeling the xylem of isolated stem segments of L.lancifolium was solution soaking for 24 h.The running speed of CF in the isolated stem was 0.3 cm/h,which was consistent with the running speed of the material in the field.CF could be transported between the xylem and parenchyma cells,indicating that the material transport in the xylem could be through the symplastic pathway.The above results laid a foundation for the study of the xylem transport mechanism and the xylem pathogen disease of lily.展开更多
We investigated the osmotic stress and ion-specific effects on xylem abscisic acid (ABA), ion uptake and transport and gas exchange in one-year-old seedlings of Populus euphratica Oliv. Net photosynthetic rates (P-n) ...We investigated the osmotic stress and ion-specific effects on xylem abscisic acid (ABA), ion uptake and transport and gas exchange in one-year-old seedlings of Populus euphratica Oliv. Net photosynthetic rates (P-n) and unit transpiration rates (TRN) were both significantly decreased upon an osmotic shock caused by PEG 6000 solution (osmotic potential = -0.24 MPa) or a saline, which was applied by 50 mmol/L Na+-salts (NaNO3 : NaHCO3 : NaH2PO4 = 5 : 4 : 1, pH 6.8, osmotic potential = -0.24 MPa) or by 50 mmol/L Cl--salts (KCl : NH4Cl = 1:1, osmotic potential = -0.24 MPa). However, salt-treated P. euphratica plants maintained typically higher TRN than those exposed to PEG. Xylem ABA concentrations increased rapidly following the PEG treatment, exhibiting peaking values at 1 h, then returning to pre-stress levels, followed by a gradual increase. Similarly, both Na+-treated and Cl--treated trees exhibited a rapid rise of ABA after salt stress was initiated. Notably, salt-treated plants maintained a relatively higher ABA than PEG-treated plants in a longer term. Collectively, results suggest that osmotic stress and ion-specific effects were both responsible for salt-induced ABA in P. euphratica : the initial rapid increase of xylem ABA appears to be a consequence of an osmotic shock, whereas specific salt effects seem to be responsible for ABA accumulation later on. Compared with Cl--treated trees, a higher inhibitory effect on gas exchange (P-n and TRN) was observed in Na+-salt plants, resulting from its long-sustained ABA and higher salt concentrations in the xylem. Displacement of membrane-associated Ca2+ by Na+ and the lesser capacity in Na+ compartmentation in root vacuoles likely contribute to the high influx of Na+ and Cl- in Na+-treated plants. Xylem K+, Ca2+ and Mg2+ concentrations were elevated by external Na+ -salts and Cl--salts, suggesting that P. euphratica maintained a higher capacity in nutrient uptake under saline conditions, which makes a contribution to its salinity tolerance.展开更多
The dynamic changes in the distribution of lignin and hemicelluloses (xylans and xyloglucans) in cell walls during the differentiation of secondary xylem in Eucommia ulmoides Oliv. were studied by means of ultraviolet...The dynamic changes in the distribution of lignin and hemicelluloses (xylans and xyloglucans) in cell walls during the differentiation of secondary xylem in Eucommia ulmoides Oliv. were studied by means of ultraviolet light microscopy and transmission electron microscopy combined with immunogold labelling. In the cambial zone and cell expansion zone, xyloglucans were localized both in the tangential and radial walls, but no xylans or lignin were found in these regions. With the formation of secondary wall S-1 layer, lignin occurred in the cell corners and middle lamella, while xylans appeared in S-1 layer, and xyloglucans were localized in the primary walls and middle lamella. In pace with the formation of secondary wall S-2 and S-3 layer, lignification extended to S-1, S-2 and S-3 layer in sequence, showing a patchy style of lignin deposition. Concurrently, xylans distributed in the whole secondary walls and xyloglucans, on the other hand, still localized in the primary walls and middle lamella. The results indicated that along with the formation and lignification of the secondary wall, great changes had taken place in the cell walls. Different parts of cell walls, such as cell corners, middle lamella, primary walls and various layers of secondary walls, had different kinds of hemicelluloses, which formed various cell wall architecture combined with lignin and other cell wall components.展开更多
The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation ...The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation technique. Throughout the differentiation, which is a typical programmed cell death (PCD) process, ATPase deposits increased in the nucleus but decreased and progressively disappeared in the cell organelles. At the same time, the distribution of ATPase increased in the inner face of the cell wall and pits with cytoplasmic degeneration. The results demonstrated that the PCD was an energy dependent active process and was controlled by nuclear genes. On the other hand, the distribution of ATPase in the intercellular spaces increased with the formation of the new cambium resulted from the dedifferentiation of the secondary xylem cells after girdling. However, ATPase was not found in the nucleus of the dividing cells, suggesting that nutrients were transported through protoplast during differentiation, and through both protoplast and apoplast during dedifferentiation. Thus, the energy required in cell division was provided mainly by intercellular spaces. These findings indicate that the dynamic distribution of ATPase reflected which cell component was actively taking part in the cell metabolism at various stages of the plant development, and its distribution was associated with the physiological state of the cell. Based on the characteristic distributions of ATPase, the critical stage of cell differentiation and the relationship between the critical stage and dedifferentiation were discussed.展开更多
Programmed cell death (PCD) during secondary xylem differentiation in Eucommia ulmoides Oliv. was examined using electron microscopy and by investigation of DNA fragmentation and degradation of caspase-like proteases ...Programmed cell death (PCD) during secondary xylem differentiation in Eucommia ulmoides Oliv. was examined using electron microscopy and by investigation of DNA fragmentation and degradation of caspase-like proteases (CLPs). DNA ladders were detected in developing secondary xylem by gel electrophoresis. DNA fragmentation was further confirmed by using the TdT-mediated dUTP nick-end labeling (TUNEL) method. Western blotting analysis showed that CLPs (caspase-8- and caspase-3-like proteases) and PARP (poly (ADP-ribose) polymerase) were degraded during secondary xylem differentiation. The results thus indicated that secondary xylem differentiation in E ulmoides was a typical process of PCD and the degradation of CLPs might be a constitutive PCD event during secondary xylem differentiation.展开更多
The study on the changes of stomatal sensitivity in relation to xylem ABA during periodical soil drying and the effect of leaf water status on the stomatal sensitivity has confirmed that xylem ABA concentration is a g...The study on the changes of stomatal sensitivity in relation to xylem ABA during periodical soil drying and the effect of leaf water status on the stomatal sensitivity has confirmed that xylem ABA concentration is a good indicator of soil water status around roots and the relation between xylem ABA concentration and predawn leaf water potential remained constant during the three consecutive soil drying cycles based on the slopes of the fitted lines. The sensitivity of stomata to xylem ABA increased substantially as the soil drying cycles progressed, and the xylem ABA concentration needed to cause a 50% decrease of stomatal conductance was as low as 550 nmol/L in the next two soil drying cycle, as compared with the 750 nmol/L ABA in the first cycle of soil drying. The results using the split_root system showed that leaf water deficit significantly enhanced the stomatal response to xylem ABA and the xylem ABA concentration needed to cause a 50% decrease in stomatal conductance was 2 to 4 times smaller in the whole_root_drying treatment than those in the semi_root_drying treatment. These results suggested that the sensitivity of stomata to xylem ABA concentration is not a fixed characteristic.展开更多
基金Financial support was obtained from National Natural Science Foundation of China(Grant Nos.32101549 and 32201585)Natural Science Foundation of Shandong Province,China(Grant No.ZR202112010288)+2 种基金Science&Technology Specific Projects in Agricultural High-tech Industrial Demonstration Area of the Yellow River Delta(Grant No.2022SZX39)Technical Innovation and Application Development Special Project of Chongqing(Grant No.CSTB2022TIAD-LDX0013)the Taishan Scholar Program of Shandong(Grant No.tsqn202103092)。
文摘Tyrosine decarboxylase(TyDC)converts tyrosine to tyramine and plays a crucial role in secondary metabolite reactions,development,and stress responses in plants.Currently,the biological role of TyDC proteins from trees is unknown.This study provided evidence showing that poplar PaTyDC4 functions in wood development and drought stress response.PaTyDC4 is preferentially expressed in wood-forming cells in stems.Overexpression of PaTyDC4 in poplars under the control of a xylem-specific promoter resulted in an increase in the ratio of xylem to phloem width,vessel cell area,and lignin accumulation in the stems.Biochemical assays revealed that PaTyDC4 was a component of the PaC3H17-PaMYB199 module-mediated pathway.In poplar stems,Pa TyDC4 expression was directly suppressed by PaMYB199,which was attenuated by the interaction between PaC3H17 and PaMYB199.In addition,Pa TyDC4 overexpression lines showed stronger drought tolerance than the wild-type lines,with higher photosynthetic capacity and lower levels of H_(2)O_(2).These results indicate that PaTyDC4 promotes xylem differentiation and lignin deposition during secondary growth and confers drought tolerance.Our findings may be useful for the genetic modification of biomass and drought resistance in trees.
基金supported by the Natural Science Basic Research Program of Shaanxi Province,China(2024JCYBQN-0491)Heng Wan would like to thank the Chinese Scholarship Council(CsC)(202206300064)。
文摘While biochar amendment enhances plant productivity and water-use efficiency(WUE),particularly under waterlimited conditions,the specific mechanisms driving these benefits remain unclear.Thus,the present study aims to elucidate the synergistic effects of biochar and reduced irrigation on maize(Zea mays L.)plants,focusing on xylem composition,root-to-shoot signaling,stomatal behavior,and WUE.Maize plants were cultivated in splitroot pots filled with clay loam soil,amended by either wheat-straw biochar(WSB)or softwood biochar(SWB)at 2%(w/w).Plants received full irrigation(FI),deficit irrigation(DI),or partial root-zone drying rrigation(PRD)from the 4-leaf to the grain-filling stage.Our results revealed that the WSB amendment significantly enhanced plant water status,biomass accumulation,and WUE under reduced irrigation,particularly when combined with PRD.Although reduced irrigation inhibited photosynthesis,it enhanced WUE by modulating stomatal morphology and conductance.Biochar amendment combined with reduced rrigation significantly increased xylem K^(+),Ca^(2+),Mg^(2+),NO_(3)^(-),Cl^(-),PO_(4)^(3-),and SO_(4)^(2-)-but decreased Na+,which in turn lowered xylem pH.Moreover,biochar amendment and especially WSB amendment further increased abscisic acid(ABA)contents in both leaf and xylem sap under reduced irrigation conditions due to changes in xylem ionic constituents and pH.The synergistic interactions between xylem components and ABA led to refined adjustments in stomatal size and density,thereby affecting stomatal conductance and ultimately improving the WUE of maize plants at different scales.The combined application of WSB and PRD can,therefore,emerge as a promising approach for improving the overall plant performance of maize plants with increased stomatal adaptations and WUE,especially under water-limited conditions.
基金supported by the Natural Science Foundation of China (project number 31861133008)financial support from the Deutsche Forschungsgemeinschaft (German Research Foundation, DFG, project number 410768178)
文摘Lipids may play an important role in preventing gas embolisms by coating nanobubbles in xylem sap.Few studies on xylem sap lipids have been reported for temperate plants,and it remain unclear whether sap lipids have adaptational significance in tropical plants.In this study,we quantify the lipid composition of xylem sap for angiosperm species from a tropical savanna(seven species)and a seasonal rainforest(five species)using mass spectrometry.We found that all twelve species studied contained lipids in their xylem sap,including galactolipids,phospholipids and triacylglycerol,with a total lipid concentration ranging from 0.09 to 0.26 nmol/L.There was no difference in lipid concentration or composition between plants from the two sites,and the lipid concentration was negatively related to species’open vessel volume.Furthermore,savanna species showed little variation in lipid composition between the dry and the rainy season.These results support the hypothesis that xylem sap lipids are derived from the cytoplasm of individual conduit cells,remain trapped inside individual conduits,and undergo few changes in composition over consecutive seasons.A xylem sap lipidomic data set,which includes 12 tropical tree species from this study and 11 temperate tree species from literature,revealed no phylogenetic signals in lipid composition for these species.This study fills a knowledge gap in the lipid content of xylem sap in tropical trees and provides additional support for their common distribution in xylem sap of woody angiosperms.It appears that xylem sap lipids have no adaptive significance.
基金the National Natural Science Foundation of China(31902043,32172612).
文摘The xylem undergoes physiological changes in response to various environmental conditions during the process of plant growth.To understand these physiological changes,it is extremely important to observe the transport of xylem.In this study,the distribution and structure of vascular bundle in Lilium lancifolium were observed using the method of semithin section.Methods for introducing a fluorescent tracer into the xylem of the stems were evaluated.Then,the transport rule of 5(6)-Carboxyfluorescein diacetate(CFDA)in the xylem of the stem of L.lancifolium was studied by fluorescence dye in live cells tracer technology.The results showed that the vascular bundles of L.lancifolium were scattered in the basic tissue,the peripheral vascular bundles were smaller and densely distributed,and the closer to the center,the larger the volume of vascular bundles and the more sparsely distributed.The vascular bundles of L.lancifolium are limited external tenacity vascular bundles,which are composed of phloem and xylem.The most suitable method for CFDA labeling the xylem of isolated stem segments of L.lancifolium was solution soaking for 24 h.The running speed of CF in the isolated stem was 0.3 cm/h,which was consistent with the running speed of the material in the field.CF could be transported between the xylem and parenchyma cells,indicating that the material transport in the xylem could be through the symplastic pathway.The above results laid a foundation for the study of the xylem transport mechanism and the xylem pathogen disease of lily.
文摘We investigated the osmotic stress and ion-specific effects on xylem abscisic acid (ABA), ion uptake and transport and gas exchange in one-year-old seedlings of Populus euphratica Oliv. Net photosynthetic rates (P-n) and unit transpiration rates (TRN) were both significantly decreased upon an osmotic shock caused by PEG 6000 solution (osmotic potential = -0.24 MPa) or a saline, which was applied by 50 mmol/L Na+-salts (NaNO3 : NaHCO3 : NaH2PO4 = 5 : 4 : 1, pH 6.8, osmotic potential = -0.24 MPa) or by 50 mmol/L Cl--salts (KCl : NH4Cl = 1:1, osmotic potential = -0.24 MPa). However, salt-treated P. euphratica plants maintained typically higher TRN than those exposed to PEG. Xylem ABA concentrations increased rapidly following the PEG treatment, exhibiting peaking values at 1 h, then returning to pre-stress levels, followed by a gradual increase. Similarly, both Na+-treated and Cl--treated trees exhibited a rapid rise of ABA after salt stress was initiated. Notably, salt-treated plants maintained a relatively higher ABA than PEG-treated plants in a longer term. Collectively, results suggest that osmotic stress and ion-specific effects were both responsible for salt-induced ABA in P. euphratica : the initial rapid increase of xylem ABA appears to be a consequence of an osmotic shock, whereas specific salt effects seem to be responsible for ABA accumulation later on. Compared with Cl--treated trees, a higher inhibitory effect on gas exchange (P-n and TRN) was observed in Na+-salt plants, resulting from its long-sustained ABA and higher salt concentrations in the xylem. Displacement of membrane-associated Ca2+ by Na+ and the lesser capacity in Na+ compartmentation in root vacuoles likely contribute to the high influx of Na+ and Cl- in Na+-treated plants. Xylem K+, Ca2+ and Mg2+ concentrations were elevated by external Na+ -salts and Cl--salts, suggesting that P. euphratica maintained a higher capacity in nutrient uptake under saline conditions, which makes a contribution to its salinity tolerance.
文摘The dynamic changes in the distribution of lignin and hemicelluloses (xylans and xyloglucans) in cell walls during the differentiation of secondary xylem in Eucommia ulmoides Oliv. were studied by means of ultraviolet light microscopy and transmission electron microscopy combined with immunogold labelling. In the cambial zone and cell expansion zone, xyloglucans were localized both in the tangential and radial walls, but no xylans or lignin were found in these regions. With the formation of secondary wall S-1 layer, lignin occurred in the cell corners and middle lamella, while xylans appeared in S-1 layer, and xyloglucans were localized in the primary walls and middle lamella. In pace with the formation of secondary wall S-2 and S-3 layer, lignification extended to S-1, S-2 and S-3 layer in sequence, showing a patchy style of lignin deposition. Concurrently, xylans distributed in the whole secondary walls and xyloglucans, on the other hand, still localized in the primary walls and middle lamella. The results indicated that along with the formation and lignification of the secondary wall, great changes had taken place in the cell walls. Different parts of cell walls, such as cell corners, middle lamella, primary walls and various layers of secondary walls, had different kinds of hemicelluloses, which formed various cell wall architecture combined with lignin and other cell wall components.
文摘The ultracytochemical localization of ATPase in the secondary xylem cells during their differentiation and dedifferentiation in the girdled Eucommia ulmoides Oliv. was carried out using a lead phosphate precipitation technique. Throughout the differentiation, which is a typical programmed cell death (PCD) process, ATPase deposits increased in the nucleus but decreased and progressively disappeared in the cell organelles. At the same time, the distribution of ATPase increased in the inner face of the cell wall and pits with cytoplasmic degeneration. The results demonstrated that the PCD was an energy dependent active process and was controlled by nuclear genes. On the other hand, the distribution of ATPase in the intercellular spaces increased with the formation of the new cambium resulted from the dedifferentiation of the secondary xylem cells after girdling. However, ATPase was not found in the nucleus of the dividing cells, suggesting that nutrients were transported through protoplast during differentiation, and through both protoplast and apoplast during dedifferentiation. Thus, the energy required in cell division was provided mainly by intercellular spaces. These findings indicate that the dynamic distribution of ATPase reflected which cell component was actively taking part in the cell metabolism at various stages of the plant development, and its distribution was associated with the physiological state of the cell. Based on the characteristic distributions of ATPase, the critical stage of cell differentiation and the relationship between the critical stage and dedifferentiation were discussed.
文摘Programmed cell death (PCD) during secondary xylem differentiation in Eucommia ulmoides Oliv. was examined using electron microscopy and by investigation of DNA fragmentation and degradation of caspase-like proteases (CLPs). DNA ladders were detected in developing secondary xylem by gel electrophoresis. DNA fragmentation was further confirmed by using the TdT-mediated dUTP nick-end labeling (TUNEL) method. Western blotting analysis showed that CLPs (caspase-8- and caspase-3-like proteases) and PARP (poly (ADP-ribose) polymerase) were degraded during secondary xylem differentiation. The results thus indicated that secondary xylem differentiation in E ulmoides was a typical process of PCD and the degradation of CLPs might be a constitutive PCD event during secondary xylem differentiation.
文摘The study on the changes of stomatal sensitivity in relation to xylem ABA during periodical soil drying and the effect of leaf water status on the stomatal sensitivity has confirmed that xylem ABA concentration is a good indicator of soil water status around roots and the relation between xylem ABA concentration and predawn leaf water potential remained constant during the three consecutive soil drying cycles based on the slopes of the fitted lines. The sensitivity of stomata to xylem ABA increased substantially as the soil drying cycles progressed, and the xylem ABA concentration needed to cause a 50% decrease of stomatal conductance was as low as 550 nmol/L in the next two soil drying cycle, as compared with the 750 nmol/L ABA in the first cycle of soil drying. The results using the split_root system showed that leaf water deficit significantly enhanced the stomatal response to xylem ABA and the xylem ABA concentration needed to cause a 50% decrease in stomatal conductance was 2 to 4 times smaller in the whole_root_drying treatment than those in the semi_root_drying treatment. These results suggested that the sensitivity of stomata to xylem ABA concentration is not a fixed characteristic.