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In rice (Oryza sativa L.), the maintenance of high photosynthetic rate of flag leaves and the carbon remobilization from leaf sheaths after heading is a critical physiological component affecting the yield. To clarify the genetic basis of RuBisCO content of the flag leaf, a major determinant of photosynthetic rate, and non-structural carbohydrate (NSC) concentration in the third leaf sheath at heading, we carried out quantitative trait loci (QTL) analysis with 39 Koshihikari/Kasalath chromosome segment substitution lines (CSSLs) and backcross progeny F2 population derived from target CSSL holding the QTL/Koshihikari in the field. QTLs for RuBisCO content and NSC concentration at heading were detected between R2447-C1286 and R2447-R716 on chromosome 10, respectively, by comparing Koshihikari with four CSSLs for chromosome 10 (SL-229, -230, -231 and -232). The progeny QTL for RuBisCO content and for NSC concentration at heading qRCH-10 and qNSCLSH-10-1, respectively, were detected at similar marker intervals between RM8201 and RM5708. In addition, QTLs for RuBisCO content at 14 d after heading, qRCAH-10-1 and qRCAH-10-2, were detected in regions different from that of qRCH-10. No QTL for NSC concentration at 14 d after heading was detected between RM8201 and R716, the region analyzed in this study. The QTLs qRCH-10 and qRCAH-10-1 for RuBisCO content would have additive effects. These QTLs for RuBisCO content and NSC concentration newly found using CSSLs and their backcross progeny F2 population should be useful for better understanding the genetic basis of source and temporary-sink functions in rice and for genetic improvement of Koshihikari in terms of their functions.

Grain filling of rice (Oryza sativa L.) is thought to be affected not only by the supply of carbohydrate from source organs, but also by sink activity, including the activities of enzymes related in starch synthesis. To examine the effect of translocated carbohydrate on sink activity, we cultured detached ears (cut 7 days after flowering) in a solution containing sucrose at various concentrations, glutamine and vitamins. Grain weights 16 days after the start of ear culture increased with increasing sucrose concentration up to 50-70 g sucrose l -1 , but not above this range, probably because of low water potential. Activities of ADP glucose pyrophosphorylase, soluble starch synthase and sucrose synthase rapidly decreased in grains grown without sucrose, but increased with increasing sucrose concentration up to 50-70 g l -1 . Expression of mRNAs for these enzymes and for sucrose transporter was also generally increased by sucrose at 50-70 g l -1 . Though there was a good correlation between activity and expression of mRNA for sucrose synthase, the correlation for other enzymes was not very good. Thus, we suggested that the sink activity of grains in rice ears, and therefore the grain filling, was affected by the supply of carbohydrate from source organs, and that it was also owing to factors other than transcriptional regulation.

In rice, caryopses located at the base of the panicle have a lower growth rate than those at the tip of the panicle. The former and latter types of caryopses are called inferior and superior caryopses, respectively. Taking the different growth rate into consideration, sugar status and the expression of genes encoding carbohydrate-metabolizing enzymes in inferior caryopses were compared with those in superior caryopses. During the first 5 d after flowering, superior caryopses elongated rapidly, but inferior caryopses did not. At this phase, inferior caryopses had a low ratio of hexose to sucrose, high activity of acid invertase and the absence of the expression of the genes encoding the above enzymes except for two isoforms of cell wall invertase, OsCIN4 and INV1, in comparison with superior caryopses. At the start of caryopsis elongation in both superior and inferior caryopses, the hexose/sucrose ratio increased accompanied by gene expression of vacuolar invertase (INV3), sucrose synthase (RSus1) and ADP-glucose pyrophosphorylase (AGP-L2: D50317). Furthermore, the genes related to endospermal starch accumulation were expressed highly with the decrease in the hexose/sucrose ratio after its peak. Based on the comparison of superior and inferior caryopses, the possible mechanism of grain filling in rice is discussed.

Carbohydrate contents varied with position in a leaf sheath, and differed between the flag leaf sheath and the second leaf sheath below the flag leaf (-2 leaf sheath) in rice (Oryza sativa L.). In the -2 leaf sheath before heading, light microscopy revealed differences in the distribution of starch granules depending on position. Leaf sheaths were divided into several parts, and the contents of carbohydrates (starch, sucrose, and hexoses) were measured in each part. Before heading, the content of accumulated starch increased linearly from the top to the bottom in -2 leaf sheaths (r2=0.99, P

To investigate whether increased sucrose-phosphate synthase (SPS) activity alters photosynthetic activity and/or the export of carbon from leaves under elevated C02 partial pressure ([C02]), we raised two lines of transgenic rice (H54-9 and H69-7), each overexpressing a maize SPS gene, and wild-type rice under ambient [C02] (35 Pa) and elevated [0O2] (100 Pa). Under ambient [C02], no significant difference was observed between the transgenic and wild-type plants in the levels of sucrose or starch in leaves or the photosynthetic activity; but the carbon export rate was higher in H69-7 than in the wild-type. Under elevated [C02], photosynthetic activity increased in all plants, but the accumulation of starch was significantly repressed in H54-9, whose SPS activity was about 12.5 times higher than that of the wild-type. The carbon export rate was higher in both transgenic lines than the wild-type. We considered that increased SPS activity in rice plants would promote the export of carbon from leaves and, as a result, starch accumulation in the leaves would be suppressed and/or photosynthetic activity would be promoted under elevated [CO2].

It has been reported that carbonic anhydrase (CA) activity in plant leaves is decreased by Zn deficiency. We examined the effects of Zn deficiency on the activity of CA and on photosynthesis by leaves in rice plants (Oryza sativaL.). Zn deficiency increased the transfer resistance from the stomatal cavity to the site of CO2 fixation 2.3-fold and, consequently, the value of the transfer resistance relative to the total resistance in the CO2-assimilation process increased from 10% to 21%. This change led to a reduced CO2concentration at the site of CO2 fixation, resulting in an increased gradient of CO2 between the stomatal cavity and this site. The present findings support the hypothesis that CA functions to facilitate the supply of CO2 from the stomatal cavity to the site of CO2 fixation. We also showed that the level of mRNA for CA decreased to 13% of the control level during Zn deficiency. This decrease resembled the decrease in CA activity, suggesting the possible involvement of the CA mRNA level in the regulation of CA activity.

Our previous paper showed that cultivar difference of flag leaf photosynthesis (LPS) in rice (Oryza sativa L.) was attributed to the difference in mesophyll resistance (rm). In this paper, we tried to divide rm into CO2 transfer resistance (rr) and CO2 fixation resistance (rc) for further analysis of rm by means of 3C determination of photosyntheti c products. In the heading time, rr, and rc occupied 14, and 58% in the total resistance, respectively, in average through 31 cultivars. This suggests that cultivar difference of LPS is mainly caused by the difference of r c. The percentage ratios of each resistance to the total one were maintained comparatively constant through the senescing process of the flag leaves. Moreover, the CO2 concentration in CO 2 fixating site showed almost the same values in different cultivars in a whole life span of a flag leaf. The rc showed a close correlation with ribulose-l,5-bisphosphate carboxylase/oxygenase (RubisCO) content. Carbonic anhydrase (CA) activity showed no correlation with rr, but showed positive correlation with rc. It was suggested that a mutual regulation system between CA activity and RubisCO activity could exist.

Tropospheric ozone is an air pollutant harmful to plants and animals. Its rapid increase at the ground surface has raised serious concern over damage to the quality and yield of agricultural crops. Reactive oxygen species (ROS) are formed in plant cells when that are exposed to a high concentration of ozone, and the ROS are thought to alter gene expression and result in cellular death. Clarification of the ROS formation will provide us with a better understanding of the physiological responses to and signaling pathways of ozone stress in plants. In this study, we investigated the mechanisms of the ROS formation in rice (Oryza sativa L.), the premier crop in Asia. To determine ROS distribution in rice leaves under acute ozone stress, we analyzed superoxide dismutase (SOD) expression, lipid peroxidation, NADPH oxidase activity, and in vivo H2O2 formation. Interestingly, chloroplastic, peroxisomal and mitochondrial SODs down-regulated their expression levels under ozone stress, whereas cytosolic SODs maintained their expression level. Higher lipid peroxidation occurred after the end of ozone exposure, which suggests lipid peroxidation may not be due to ozone directly, but rather to metabolic changes caused by the ozone exposure. Activity of NADPH oxidase did not show significant change. The in vivo analysis indicated accelerated formation of H2O2 about 24 h after the onset of exposure, which suggests that cellular death occurred around this time.

Reduction of relative growth rate (RGR) due to the NaCl treatment was examined in the seedlings of six Korean rice cultivars which had been reported to be different in salt tolerance on the level of grain yield. On the level of RGR, three of the cultivars were identified as salt tolerant, and the remaining three were identified as the sensitive cultivars. Further analysis of RGR indicated that the reduction of RGR by NaCl treatment was attributed in large part to the reduction of the net assimilation rate (NAR) rather than to that of the leaf area ratio (LAR). Leaf photosynthesis which is closely related with NAR, was also reduced by NaCl treatment in salt sensitive cultivars to a greater extent than tolerant cultivars. Ribulose-1, 5-bisphosphate carboxylase/oxygenase (Rubisco) content was determined in a tolerant cultivar, Han-Kang-Chal, and a sensitive one, Dong-Hae. Han-Kang-Chal showed a smaller decrease of Rubisco content per absorbed Na in the NaCl-treated leaves. This indicates that degradation of Rubisco by absorbed Na is slower in tolerant cultivars than in sensitive cultivars. Moreover, Han-Kang-Chal showed a larger decrease in osmotic potential in the NaCl treated leaves, suggesting that osmotic adjustment was developed under salt stress conditions in a tolerant cultivar.

Carbonic anhydrase (CA; EC.4.2.1.1) catalyzes the reversible conversion of CO2 to HCO3. CA has been recognized as an important enzyme that is closely associated with photosynthesis and the activity of CA can be specifically inhibited by zinc deficiency. In a previous study, we succeeded in estimating the magnitude of the CO2 transfer resistance (rr) from the intercellular space of mesophyll cells to the site of CO2 fixation in the chloroplasts (1). The purpose of the present study was to elucidate the relationship between the activity of CA and each component of mesophyll resistance and the mechanism of the specific inhibition of CA activity by using zinc deficient rice plants.