1. Introduction
There is evidence suggesting that in plants photosynthetic matter production is regulated by photosynthetic source-sink balance, i.e., the ratio of photosynthetic source organs (e.g., leaves) to non-photosynthetic sink organs (e.g., roots) and/or the balance of supply and demand of photosynthetic carbohydrate(s) within the plant (Kasai, 2008, 2011). Plant photosynthetic dry matter production is the source of a variety of metabolic and structural compounds. Because of increasing population, shortages of energy and food may become more severe (von Caemmerer & Evans, 2010; Raines, 2011). Plant photosynthetic dry matter production is also essential for maintaing environmental quality. For example, a well-known environmental problem is climatic warming of the earth, which mainly comes from deforestation (Brovvkin et al., 2004). Improvement of plant dry matter productivity may be an effective way for solving the problems of energy, foods and climatic warming. Thus, it is important to elucidate the mechanism(s) of regulation of plant photosynthetic matter production through photosynthetic source-sink balance.
Data from a number of studies including field investigations implicate that in plants, accumulation of photosynthetic carbohydrate(s) in leaves, which occurs when photosynthetic source capacity exceeds sink capacity, can regulate leaf photosynthetic rate (Sawada et al., 1999; Kasai, 2008, 2011; Kasai et al., 2012). In soybean a significant negative correlation exists between leaf photosynthetic carbohydrate (sucrose or starch) content and photosynthetic rate (Sawada et al., 1986, 2001; Kasai, 2008). There have also been findings of photosynthetic carbohydrate-mediated decrease in the activity or the amount of Rubisco, the CO2-fixing enzyme in leaves (Sage et al., 1989; Xu et al., 1994; Martin et al., 2002; Paul & Pellny, 2003), although the detailed mechanism(s) is still unclear. To date, many studies have focused on photosynthetic carbohydrate-mediated inhibition of leaf photosynthesis to elucidate the mechanism(s) of regulation of photosynthetic matter production through photosynthetic source-sink balance. However, in contrast, there is also evidence suggesting that leaf photosynthetic rate is not necessarily affected by accumulated photosynthetic carbohydrate(s) in leaf (Nebauer et al., 2011). Apart from the regulation of leaf photosynthesis through levels of photosynthetic carbohydrate(s), it is important to examine the mechanism(s) of regulation of photosynthetic dry matter production through photosynthetic source-sink balance by focusing on new enzyme(s) thought to be important.
Data from recent studies implicate that in plants, activity(ies) of membrane H+ pump(s) such as tonoplast H+ pump(s) can be important in the regulation of photosynthetic dry matter production through photosynthetic source-sink balance (Kasai &Muto, 1990.; Schumacher et al., 1999; Li et al., 2005; Yang et al., 2007; Wang et al., 2011). However, the effect of photosynthetic source-sink balance on the activity(ies) of membrane H+ pump(s) has not been investigated. We show here experimental data of our recent study relating to this subject. We investigated in soybean plants how removal of pods, which decreases the ratio of sink to source organs, affects various characteristics related to photosynthetic dry matter production. Factors studied were leaf photosynthetic rate, stomatal conductance, transpiration rate and intercellular CO2 concentration, initial and total activities of Rubisco, chlorophyll, total protein, inorganic phosphate, photosynthetic carbohydrates (sucrose and starch), and dry weights of source and sink organs. We also investigated the effect of pod removal on activities of the H+ pumps of the leaf plasma membrane (H+-ATPase) and tonoplast (H+-ATPase and H+-PPase). It is now well known that soybean is one of the most important crops grown in the world (Board & Kahlon, 2011; Ainsworth et al., 2012). On the basis of our experimental data and the other relevant information, we also consider how membrane H+ pump(s) can be important in the regulation of photosynthetic dry matter production through photosynthetic source-sink balance.
2. Materials and methods
2.1. Plant materials
Soybean (
2.2. Leaf photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 concentration
Leaf photosynthetic rate, transpiration rate, stomatal conductance and intercellular CO2 concentration were determined on day 3 after pod removal in fully expanded middle trifoliate leaves at a light intensity of 800 µmol photons m-2 s-1, air flow rate of 200 ml min-1, air temperature of 25 oC, relative humidity of 60 % and CO2 concentration of 350 ppm using a portable photosynthetic analyzer (Cylus-1; Koito Industries Ltd.). After measurements, leaf disks (1.79 cm2) were taken from the middle trifoliate leaves for the other analyses (see 2.3), as described previously (Kasai, 2008).
2.3. Rubisco activity, chlorophyll, protein, phosphate, sucrose and starch
Initial and total activities of Rubisco in leaf extract were determined at 25 oC as described previously (Kasai, 2008). Leaf chlorophyll content was determined according to the method of Mackinney (1941). Leaf total protein content was determined by quantifying protein included in the leaf extract that had been prepared for determination of Rubisco activity by the method of Bradford (1976). Leaf inorganic phosphate content was determined according to the method of Saheki et al. (1985). Leaf sucrose and starch contents were determined as described by Sawada et al. (1995).
2.4. Dry weight
For determination of dry weights of source (leaves) and sink organs (stems, floral organs including pods, and roots), organs were separated from plants on day 3 after pod removal and dried at 70oC for a week.
2.5. Plasma membrane, tonoplast and H+ pump activity
The activities of H+ pumps of leaf plasma membrane and tonoplast were determined using plasma membrane vesicles and tonoplast vesicles prepared from leaves. Plasma membrane vesicles and tonoplast vesicles were prepared from leaves (25 g in fresh weight) of plants on day 3 after pod removal essentially as described by Nouri & Komatsu (2010) and Maeshima & Yoshida (1989), respectively. For preparation of plasma membrane vesicles, leaf-homogenizing medium consisted of 0.3 M sucrose, 50 mM Tris, 8 mM EDTA (acid form), 2 mM PMSF, 4 mM DTT and 0.2 % (w/v) BSA, and its volume was 200 ml. After homogenization, the medium was filtered through four layers of gauze and the filtrate was centrifuged at 10,000 g for 20 min. After supernatant was centrifuged at 80,000 g for 40 min, the pellets were suspended with a sucrose-containing medium [0.3 M sucrose, 5 mM KH2PO4, 5 mM KCl, 0.1 mM EDTA and 0.1 mM DTT (pH 7.8)], of which volume was 10 ml. The plasma membrane vesicles were prepared from the suspension by using aqueous two-phase partitioning method. Dilution of the upper layers that had been obtained was conducted with a sorbitol-containing medium [0.25 M sorbitol, 5 mM HEPES-BTP and 0.1 mM DTT (pH 7.0)]. The final pellets of the plasma membrane vesicles after centrifugation (80,000 g, 40 min) were suspended with another sorbitol-containing medium [1 M sorbitol, 5 mM HEPES-BTP and 0.1 mM DTT (pH 7.0)] and stored at -80 oC until uses. For preparation of tonoplast vesicles, leaf-homogenizing medium consisted of 0.25 M sorbitol, 50 mM HEPES-KOH, 5 mM EGTA, 1 mM PMSF, 2.5 mM Na2S2O5 and 1.5% (w/v) PVP (pH 7.6), and its volume was 200 ml. After homogenization, the medium was filtered through four layers of gauze and the filtrate was centrifuged at 4000 g for 10 min. After supernatant was centrifuged at 80,000 g for 60 min, the pellets were suspended with a sucrose-containing medium [0.3 M sucrose, 10 mM KH2PO4, 1 mM EGTA and 2 mM DTT (pH 7.8)], of which volume was 5 ml, and 3 ml of a sorbitol-containing medium [0.25 M sorbitol, 50 mM HEPES-KOH, 1 mM EGTA and 2 mM DTT (pH 7.3)] was put on the suspension and the solution was centrifuged (120,000 g, 40 min). The resulting middle layer was diluted with the same sorbitol-containing medium. The final pellets of the tonoplast vesicles after centrifugation (150,000 g, 20 min) were suspended with another sorbitol-containing medium [0.25 M sorbitol, 5 mM HEPES-BTP, 2 mM DTT (pH 7.5)] and stored at -80oC until uses.
The activity of H+ pump, i.e., H+-ATPase of leaf plasma membrane was determined at 30oC as vanadate-sensitive ATP-hydrolytic activity (kasai & Sawada, 1994). The activities of H+ pumps, i.e., H+-ATPase and H+-PPase of leaf tonoplast were determined at 30oC as nitrate-sensitive ATP-hydrolytic activity and Na+-sensitive PPi-hydrolytic activity (Kasai et al., 1993; Kasai & Sawada, 1994), respectively. Reaction medium (500 μl) for the activity of plasma membrane H+ pump consisted of 50 mM HEPES-BTP (pH 7.0), 3 mM MgSO4, 3 mM ATP, 1 mM EGTA, 50 mM KCl, ± 0.1 mM Na3VO4, 0.02% (w/v) Triton X-100 and membrane vesicles (10 μg). Reaction medium for the activity of tonoplast H+-ATPase consisted of ±50 mM HEPES-BTP (pH 7.5), 3 mM MgSO4, 3 mM ATP, 1 mM EGTA, ±50 mM KCl, ± 50 mM KNO3, 0.02% (w/v) Triton X-100 and membrane vesicles (10 μg). Reaction medium for the activity of tonoplast H+-PPase consisted of 50 mM HEPES-BTP (pH 7.5), 5 mM MgSO4, 0.5 mM PPi, 1 mM EGTA, 50 mM KNO3, ± 50 mM NaNO3, 0.02% (w/v) Triton X-100 and membrane vesicles (10 μg). Phosphate liberated from substrate ATP or PPi was determined according to the method of Saheki et al. (1985).
3. Results
Analyzed leaf photosynthetic rate and transpiration rate were significantly lower in depodded plants than in control plants (Fig. 1). Leaf stomatal conductance was also lower in depodded plants than in control plants, while leaf intercellular CO2 concentration did not differ significantly between control and depodded plants (Fig. 2). Initial and total activities of Rubisco in leaf extract did not differ significantly between control and depodded plants (Fig. 3). Contents of chlorophyll, total protein and inorganic phosphate in leaves were all significantly higher in depodded plants than in control plants (Fig. 4). Contents of sucrose and starch in leaves did not differ significantly between control and depodded plants (Fig. 5). Activity of H+ pump (H+-ATPase) of leaf plasma membrane and activities of H+ pumps (H+-ATPase and H+-PPase) of leaf tonoplast were all significantly lower in depodded plants than in control plants (Fig. 6). Dry weights of leaves, stems and roots did not differ significantly between control and depodded plants (Fig. 7). When the ratio of sink (stems + floral organs including pods + roots) to source organs (leaves) was calculated, those in control and depodded plants were on the average 1.25 (100%) and 0.70 (56%), respectively.
4. Discussion
As described in the Introduction, it is important to examine the mechanism(s) of regulation of plant photosynthetic dry matter production through photosynthetic source-sink balance by focusing on a new enzyme(s). We focused on H+ pumping enzymes of leaf plasma membrane (H+-ATPase) and tonoplast (H+-ATPase and H+-PPase), and investigated in soybean plants how removal of pods, which decreases the ratio of sink to source organs, affects various characteristics related to photosynthetic dry matter production and the activities of H+ pumps. Pod removal was shown to decrease largely leaf photosynthetic rate, transpiration rate and stomatal conductance without affecting significantly leaf intracellular CO2 concentration (Fig. 1 and 2). These results imply that pod removal decreased equally the rate of CO2 diffusion via leaf stomata and the rate of CO2 fixation in leaf photosynthetic cells. In plants, Rubisco is a major protein in leaves (Furbank et al., 1996; von Caemmerer et al., 2005), and there is evidence from studies altering the expressions of Rubisco or its activation enzyme, Rubisco activase, that changes in the activity or the amount of Rubisco in leaves significantly affect leaf photosynthetic rate (Furbank et al., 1996; von Caemmerer et al., 2005). There is also a report demonstrating that a rough and positive correlation exists between leaf chlorophyll content and photosynthetic rate (Arp, 1991). Therefore, it is speculated that the pod removal-induced decrease in leaf photosynthetic rate might have resulted from a decrease in the activity or the amount of Rubisco in the leaf or the content of leaf chlorophyll. However, data of Figure 3 and 4 indicate that pod removal did not significantly affect potential activity of Rubisco in the leaf and could not decrease the content of Rubisco or chlorophyll, suggesting that the pod removal-induced decrease in leaf photosynthetic rate did not result from either of these factors.
Previously, in single-rooted soybean leaves that are the same species as we used in our pod removal study, it was demonstrated that a decrease in leaf inorganic phosphate content can result in a decrease in leaf Rubisco activity in vivo (Sawada et al., 1990, 1992). In vitro, inorganic phosphate has been found to promote the binding of activator CO2 to uncarbamylated inactive Rubisco (Bhagwat, 1981; McCurry et al., 1981; Anwaruzzaman et al., 1995). Data of Figure 4 indicate that pod removal did not decrease leaf inorganic phosphate content. This result indicates that the pod removal-induced decrease in leaf photosynthetic rate did not result from a decrease in leaf inorganic phosphate content.
There is a hypothesis of inhibition of photosynthesis through accumulation of sucrose in the leaf, although the detailed mechanism(s) is still unclear (Kasai, 2008). For example, in a study having continuous exposure to light of single-rooted soybean leaves, a significant negative correlation was shown between leaf sucrose content and photosynthetic rate (Sawada et al., 1986). It is thought that in both control and depodded plants, sucrose-induced inhibition of leaf photosynthesis was, if any, very small. Leaf sucrose content of control plants, which was higher on the average than that of depodded plants (Fig. 5), corresponded with a content that led to a very small decrease in leaf photosynthetic rate of single-rooted soybean leaves (Sawada et al., 1986). Leaf sucrose content of control plants did not seem to decrease leaf photosynthetic rate in our previous pod removal study using soybean plants (Kasai et al., 2008). There is also a hypothesis of inhibition of photosynthesis through accumulation of starch in leaves (Kasai, 2008). In the same study, continuous exposure to light of single-rooted soybean leaves resulted in a significant negative correlation between leaf starch content and photosynthetic rate (Sawada et al., 1986). Another study using single-rooted soybean leaves demonstrated that accumulation of starch decreases the rate of CO2 diffusion (Sawada et al., 2001; Kasai et al., 1996). In our pod removal study, pod removal did not significantly affect leaf starch content (Fig. 5). Therefore, it is suggested that the pod removal-induced decrease in leaf photosynthetic rate did not result from accumulation of sucrose or starch in leaves.
It is believed that in plants, P-type H+ pump (H+-ATPase) exists in the plasma membrane, and V-type H+ pumps (H+-ATPase and H+-PPase) exist in the tonoplast (Hall & Williams, 1991; Barkla et al., 2008). In plant leaves, a decrease in H+ pump activity of the guard cell plasma membrane can induce decreases of stomatal conductance and transpiration rate by inducing a decrease in stomatal pore size (Tominaga et al., 2001). Although we did not analyze the H+ pump activity of the guard cell plasma membrane, it was shown that pod removal largely decreased the H+ pump activity of the leaf plasma membrane (Fig. 6). Essentially, almost the same method is used for isolation of the plasma membrane from leaves and guard cells (Becker et al., 1993). Therefore, it is suggested that a large decrease in H+ pump activity of the guard cell plasma membrane could cause the pod removal-induced decreases of leaf stomatal conductance and transpiration rate. In plant cells, a decrease in H+ pump activity of the plasma membrane can induce a decrease in the electrochemical potential difference of H+ across the plasma membrane (Hall & Williams, 1991; Barkla et al., 2008). There is increasing evidence that depolarization-activated Ca2+ channel, Ca2+-activated anion (e.g., Cl-) channel, depolarization-activated anion channel (e.g., HCO3 -), electrogenic Ca2+/H+ antiporter (which has a stoichiometry higher than 2H+/Ca2+), and CO2-transportable and Ca2+-inhibitable water channel are present in the plant plasma membrane (Thuleau et al., 1994; Roberts, 2005; Frachisse et al., 1999, Kasai et al., 1990; Song et al., 2011; Chaumont et al., 2005). Therefore, it is speculated that the observed decrease in H+ pump activity of the leaf plasma membrane could cause the decrease in the rate of CO2 transport in leaf photosynthetic cells by inducing a depolarization of the plasma membrane, a decrease in the proton motive force across the plasma membrane and a rise of Ca2+ concentration inside the plasma membrane in the leaf photosynthetic cells. The suggestion drawn on the basis of data of Figure 1 and 2 that pod removal decreased equally the rate of CO2 diffusion via leaf stomata and the rate of CO2 fixation in leaf photosynthetic cells is roughly consistent with the above-mentioned suggestion and speculation proposing the regulation of leaf stomatal conductance, transpiration rate and CO2 transport in leaf photosynthetic cells by activity of H+ pump of leaf plasma membrane. As shown in Figure 6, pod removal also greatly decreased activities of the H+ pumps (H+-ATPase and H+-PPase) of the leaf tonoplast. There is increasing evidence that electrogenic Ca2+/H+ antiporter (which has a stoichiometry higher than 2H+/Ca2+) is also present in the plant tonoplast (Blackford et al., 1990; Mei et al., 2007). Therefore, it is speculated that a large decrease in H+ pump activity of the leaf plasma membrane and a large decrease in activities of H+ pumps of the leaf tonoplast could cause cooperatively the pod removal-induced decrease in leaf photosynthetic rate by inducing equal decreases in the rate of CO2 diffusion via leaf stomata and the rate of CO2 fixation in leaf photosynthetic cells. To verify our speculations, more evidence is needed. We emphasize, however, that until now, in similar studies other than our study, activities of the H+ pumps of plasma membrane and tonoplast have not been analyzed.
With respect to the mechanism(s) of why pod removal decreased the H+ pump activity of the leaf plasma membrane and activities of the H+ pumps of the leaf tonoplast, it is inferred that plant hormones abscisic acid and cytokinin could be involved in the mechanism(s). In general, cytokinin is known to have positive effect in synthesizing chlorophyll and protein, and its content in plant cells is known to decrease under deficiency of mineral nutrients such as P and N. In contrast, abscisic acid antagonizes the effects of cytokinin, and its content in plant cells increases under conditions of mineral nutrient deficiencies (Pozsar et al., 1967; Kusnetsov et al., 1998; Salama & Wareing, 1979; Mizrahi & Richmond, 1972; Battal et al., 2003). In our pod removal study, it was shown that pod removal, which decreased the ratio of sink to source organs (Fig. 7), increased significantly the contents of chlorophyll, total protein and inorganic phosphate in the leaf (Fig. 4), implicating that pod removal might have increased cytokinin content relative to abscisic acid content in the leaf by influencing the partitioning of mineral nutrients such as P and N within the plant. In barley, it was demonstrated that abscisic acid has stimulatory effects on activities of tonoplast H+ pumps, whereas cytokinin has opposite effects antagonizing the effects of abscisic acid (Kasai et al., 1993; Fukuda & Tanaka, 2006). In
Data from recent studies using transgenic plants and those from physiological studies implicate that in plants, activity(ies) of membrane H+ pump(s) can be important in the regulation of photosynthetic dry matter production through photosynthetic source-sink balance. For example, in a study using
5. Conclusion
Data from recent studies implicate that in plants, activity(ies) of membrane H+ pump(s) can be important in regulation of photosynthetic dry matter production through photosynthetic source-sink balance. However, the effect of photosynthetic source-sink balance on the activity(ies) of membrane H+ pump(s) has not been investigated. In our recent study, we investigated in soybean plants how pod removal, which decreases the ratio of sink to source organs, affects various characteristics related to photosynthetic matter production. We also investigated, for the first time, the effect of pod removal on activities of H+ pumps of leaf plasma membrane and tonoplast. From the data obtained and the other relevant information, it was concluded that in plants, changes in activity(ies) of membrane H+ pump(s) can actually play key roles in the regulation of photosynthetic dry matter production through photosynthetic source-sink balance, and that hormones abscisic acid and cytokinin may be involved in regulation of activities of tonoplast H+ pumps. Plant photosynthetic dry matter production is essential for all living organisms and is also essential for creating sound environments. Therefore, further studies are important to elucidate the detailed mechanism(s) of how membrane H+ pump(s) are involved in the regulation of photosynthetic dry matter production through photosynthetic source-sink balance.
References
- 1.
Ainsworth E. A. Yendrek C. R. Skoneczka J. A. Long S. P. 2012 Accelerating yield potential in soybean: potential targets for biotechnological improvement. 35 38 52 - 2.
Anwaruzzaman Sawada. S. Usuda H. Yokota A. 1995 Regulation of ribulose-1,5-bisphosphate carboxylase/oxygenase activation by inorganic phosphate through stimulating the binding of the activator CO2 to the activation sites. 36 425 433 - 3.
Arp W. J. 1991 Effects of source-sink relations on photosynthetic acclimation to elevated CO2.14 869 875 - 4.
Barkla B. J. Hirschi K. D. Pittman J. K. 2008 Exchangers man the pumps. Functional interplay between proton pumps and proton-coupled Ca2+ exchangers. 3 354 356 - 5.
Battal P. Turker M. Tileklioglu B. 2003 Effects of different mineral nutrients on abscisic acid in maize (Zea mays). 40 301 308 - 6.
Batelli G. Verslues P. E. Agius F. Qiu Q. Fujii H. Pan S. Schumaker K. S. Grillo S. Zhu J. K. 2007 SOS2 promotes salt tolerance in part by interacting with the vacuolar H+-ATPase and upregulating its transport activity. 27 7781 7790 - 7.
Becker D. Zeilinger C. Lohse G. Depta H. Hedrich R. 1993 Identification and biochemical characterization of the plasma-membrane H+-ATPase in guard cells of Vicia faba L. 190 44 50 - 8.
Bhagwat A. S. 1981 Activation of spinach ribulose-1,5-bisphosphate carboxylase by inorganic phosphate. 23 197 206 - 9.
Blackford S. Rea P. A. Sanders D. 1990 Voltage sensitivity of H+/Ca2+ antiport in higher plant tonoplast suggests a role in vacuolar calcium accumulation. 265 9617 9620 - 10.
Board J. E. Kahlon C. S. 2011 Soybean yield formation: What controls it and how it can be improved. In , H. A. El-Shemy, Ed.,1 36 InTech Open Access Publisher, Rijeka, Croatia. - 11.
Bradford M. M. 1976 A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. 72 248 254 - 12.
Chaumont F. Moshelion M. Daniels M. J. 2005 Regulation of plant aquaporin activity. 97 749 764 . - 13.
Brovvkin V. Sitch S. von Bloh. W. Claussen M. Bauer E. Cramer W. 2004 Role of land cover changes for atmospheric CO2 increase and climate change during the last 150 years. 10 1253 1266 - 14.
Frachisse J. M. Thomine S. Colcombet J. Guern J. Barbier-Brygoo H. 1999 Sulfate is both a substrate and an activator of the voltage-dependent anion channel of Arabidopsis hypocotyl cells. 121 253 261 - 15.
Fukuda A. Tanaka Y. 2006 Effects of ABA, auxin, and gibberellin on the expression of genes for vacuolar H+inorganic pyrophosphatase, H+ATPase subunit A, and Na+/H+ antiporter in barley. 44 351 358 - 16.
Furbank R. T. Chitty J. A. von Caemmerer. S. Jenkins C. L. D. 1996 Antisense RNA inhibition of RbcS gene expression reduces Rubisco level and photosynthesis in the C4 plant Flaveria bidentis. 111 725 734 - 17.
Gonzalez N. Bodt S. D. Sulpice R. Jikumaru Y. Chae E. Dhondt S. Daele T. V. Milde L. D. Weigel D. Kamiya Y. Stitt M. Beemster G. T. S. Inze D. 2010 Increased leaf size: different means to an end. 153 1261 1279 - 18.
Hall J. L. Williams L. E. 1991 Properties and functions of proton pumps in higher plants. 32 339 351 - 19.
Huertas R. Olias R. Eljakaoui Z. Galvez F. J. Li J. Morales P. A. D. Belver A. Rodriguez-Rosales M. P. 2012 Overexpression of SlSOS2 (SlClPK24) confers salt tolerance to transgenic tomato. 1 16 - 20.
Kasai M. 1999 Environmental stresses and plant vacuolar H+pumps. In , T. W. Ashenden et al., Eds.,97 104 Research Trends, Trivandrum, India. - 21.
Kasai M. 2008 Regulation of leaf photosynthetic rate correlating with leaf carbohydrate status and activation state of Rubisco under a variety of photosynthetic source/sink balances. 134 216 226 - 22.
Kasai M. 2011 Regulation of leaf photosynthesis through photosynthetic source-sink balance in soybean plants. In H. A. El-Shemy, Ed.,443 460 InTech Open Access Publisher, Rijeka, Croatia. - 23.
Kasai M. Muto S. 1990 Ca2+ pump and Ca2+/H+ antiporter in plasma membrane vesicles isolated by aqueous two-phase partitioning from corn leaves. 114 133 142 - 24.
Kasai M. Nakamura T. Kudo N. Sato H. Maeshima M. Sawada S. 1998 The activity of the root vacuolar H+-pyrophosphatase in rye plants grown under conditions deficient in mineral nutrients. 39 890 894 - 25.
Kasai M. Nakata H. Seino H. Kamata D. Tsukiyama T. 2008 Effect of sink-limitation on leaf photosynthetic rate and related characteristics in soybean plants. 11 223 227 - 26.
Kasai M. Sawada S. 1994 Evidence for decrease in vanadate-sensitive Mg2+-ATPase activity of higher plant membrane preparations in sucrose solution. 35 697 700 - 27.
Kasai M. Yamamoto Y. Maeshima M. Matsumoto H. 1993 Effects of in vivo treatment with abscisic acid and/or cytokinin on activities of vacuolar H+ pumps of tonoplast-enriched membrane vesicles prepared from barley roots. 34 1107 1115 - 28.
Kasai M. Koide K. Ichikawa Y. 2012 Effect of pot size on various characteristics related to photosynthetic matter production in soybean plants. 751731 7 - 29.
Kasai M. Yamaguchi A. Sawada S. 1996 The effects of CO2 on the photosynthetic fixation of CO2 and the activity of ribulose-1,5-bisphosphate carboxylase in single-rooted soybean leaves under sink-limited conditions. 37 1193 1196 - 30.
Kusnetsov V. Herrmann R. G. Kulaeva O. N. Oelmuller R. 1998 Cytokinin stimulates and abscisic acid inhibits greening of etiolated Lupinus luteus cotyledons by affecting the expression of the light-sensitive protochlorophyllide oxidoreductase. 259 21 28 - 31.
Li J. Yang H. Peer W. A. Richter G. Blakeslee J. Bandyopadhyay A. Titapiwantakun B. Undurraga S. Khodakovskaya M. Richards E. L. Krizek B. Murphy A. S. Gilroy S. Gaxiola R. 2005 Arabidopsis H+-PPase AVP1 regulates auxin-mediated organ development. 310 121 125 - 32.
Mackinney G. 1941 Absorption of light by chlorophyll solutions. 140 315 322 - 33.
Maeshima M. Yoshida S. 1989 Purification and properties of vacuolar membrane proton-translocating inorganic pyrophosphatase from mung bean. 264 20068 20073 - 34.
Martin T. Oswald O. Graham I. A. 2002 Arabidopsis seedlings growth, storage mobilization, and photosynthetic gene expression are regulated by carbon: nitrogen availability. 128 472 481 - 35.
Mc Curry S. D. Pierce J. Tolbert N. E. Orme-Johnson W. H. 1981 On the mechanism of effector-mediated activation of ribulose bisphosphate carboxylase/oxygenase. 256 6623 6628 - 36.
Mei H. Zhao J. Pittman J. K. Lachmansingh J. Park S. Hirschi K. D. 2007 In planta regulation of the Arabidopsis Ca2+/H+ antiporter CAX1. 58 3419 3427 - 37.
Mizrahi Y. Richmond A. E. 1972 Abscisic acid in relation to mineral deprivation. 50 667 670 - 38.
Nebauer S. G. Renau-Morata B. Guardiola J. L. Molina R. V. 2011 Photosynthesis down-regulation precedes carbohydrate accumulation under sink limitation in Citrus. 31 169 177 - 39.
Nouri M. Z. Komatsu S. 2010 Comparative analysis of soybean plasma membrane proteins under osmotic stress using gel-based and LC MS/MS-based proteomics approaches 10 1930 1945 - 40.
Paul M. J. Pellny T. K. 2003 Carbon metabolite feedback regulation of leaf photosynthesis and development. 54 539 547 - 41.
Peleg Z. Blumwald E. 2011 Hormone balance and abiotic stress tolerance in crop plants. 14 290 295 - 42.
Pospisilova J. 2003 Participation of phytohormones in the stomatal regulation of gas exchange during water stress. 46 491 506 - 43.
Pozsar B. I. El Hammady M. Kiraly Z. 1967 Cytokinin effect of benzyladenine: increase of nucleic acid and protein synthesis in bean leaves. 214 273 274 - 44.
Raines C. A. 2011 Increasing photosynthetic carbon assimilation in C3 plants to improve crop yield: Current and future strategies. 155 36 42 - 45.
Roberts S. K. 2006 Plasma membrane anion channels in higher plants and their putative functions in roots. 169 647 666 - 46.
Rolland F. Moore B. Sheen J. 2002 Sugar sensing and signaling in plants. S185 S205 - 47.
Sage R. F. Sharkey T. D. Seemann J. R. 1989 Acclimation of photosynthesis to elevated CO2 in five C3 species. 89 590 596 - 48.
Saheki S. Takeda A. Shimazu T. 1985 Assay of inorganic phosphate in mild pH range, suitable for measurement of glycogen phosphorylase activity. 148 277 281 - 49.
Salama A. M. S. El -D Wareing P. F. 1979 Effects of mineral nutrient on endogenous cytokinins in plants of sunflower (Helianthus annuus L.). 30 971 981 - 50.
Sawada S. Enomoto S. Tozu T. Kasai M. 1995 Regulation of the activity of ribulose-1,5-bisphosphate carboxylase in response to changes in the photosynthetic source-sink balance in intact soybean plants. 36 551 556 - 51.
Sawada S. Harada A. Asari Y. Asano S. Kuninaka M. Kawamura H. Kasai M. 1999 Effects of micro-environmental factors on photosynthetic CO2 uptake and carbon fixation metabolism in a spring ephemeral, Erythronium japonicum, growing in native and open habitats. 14 119 130 - 52.
Sawada S. Hayakawa T. Fukushi K. Kasai M. 1986 Influence of carbohydrates on photosynthesis in single, rooted soybean leaves used as a source-sink model. 27 591 600 - 53.
Sawada S. Kuninaka M. Watanabe K. Sato A. Kawamura H. Komine K. Sakamoto T. Kasai M. 2001 The mechanism to suppress photosynthesis through end-product inhibition in single-rooted soybean leaves during acclimation to CO2 enrichment. 42 1093 1102 - 54.
Sawada S. Usuda H. Hasegawa Y. Tsukui T. 1990 Regulation of ribulose-1,5-bisphosphate carboxylase activity in response to changes in the source/sink balance in single-rooted soybean leaves: the role of inorganic orthophosphate in activation of the enzyme. 31 697 704 - 55.
Sawada S. Usuda H. Tsukui T. 1992 Participation of inorganic orthophosphate in regulation of the ribulose-1,5-bisphosphate carboxylase activity in response to changes in the photosynthetic source-sink balance. 33 943 949 - 56.
Schumacher K. Vafeados D. Mc Carthy M. Sze H. Wilkins T. Chory J. 1999 The Arabidopsis det3 mutant reveals a central role for the vacuolar H+-ATPase in plant growth and development. 13 3259 3270 - 57.
Song W. Y. Choi K. S. Alexis D. A. Martinoia E. Lee Y. 2011 Brassica juncea plant cadmium resistance 1 protein (BjPCR1) facilitates the radial transport of calcium in the root. 108 19808 19813 - 58.
Thuleau P. Ward J. M. Ranjeva R. Schroeder J. I. 1994 Voltage-dependent calcium-permeable channels in the plasma membrane of a higher plant cell. 13 2970 2975 - 59.
Tominaga M. Kinoshita T. Shimazaki K. 2001 Guard-cell chloroplasts provide ATP required for H+ pumping in the plasma membrane and stomatal opening. 42 795 802 - 60.
von Caemmerer. S. Evans J. R. 2010 Enhancing C3 photosynthesis. 154 589 592 - 61.
von Caemmerer. S. Hendrickson L. Quinn V. Vella N. Millgate A. G. Furbank R. T. 2005 Reductions of Rubisco activase by antisense RNA in the C4 plant Flaveria bidentis reduces Rubisco carbamylation and leaf photosynthesis 137 747 755 - 62.
Wang L. He X. Zhao Y. Shen Y. Huang Z. 2011 Wheat vacuolar H+-ATPase subunit B cloning and its involvement in salt tolerance. 234 1 7 - 63.
Werner T. Motyka V. Laucou V. Smets R. Onckelen H. V. Schmulling T. 2003 Cytokinin-deficient transgenic Arabidopsis plants show multiple development alterations indicating opposite functions of cytokinins in the regulation of shoot and root meristem activity. 15 2532 2555 - 64.
Xu D. Q. Gifford R. M. Chow W. S. 1994 Photosynthetic acclimation in pea and soybean to high atmospheric CO2 partial pressure 106 661 671 - 65.
Yang H. Knapp J. Koirala P. Rajagopal D. Peer W. A. Silbart L. K. Murphy A. Gaxiola R. A. 2007 Enhanced phosphorus nutrition in monocots and dicots over-expressing a phosphorus-responsive type I H+-pyrophosphatase. 5 735 745