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1.  Ammonia emission from rice leaves in relation to photorespiration and genotypic differences in glutamine synthetase activity 
Annals of Botany  2011;108(7):1381-1386.
Background and Aims
Rice (Oryza sativa) plants lose significant amounts of volatile NH3 from their leaves, but it has not been shown that this is a consequence of photorespiration. Involvement of photorespiration in NH3 emission and the role of glutamine synthetase (GS) on NH3 recycling were investigated using two rice cultivars with different GS activities.
NH3 emission (AER), and gross photosynthesis (PG), transpiration (Tr) and stomatal conductance (gS) were measured on leaves of ‘Akenohoshi’, a cultivar with high GS activity, and ‘Kasalath’, a cultivar with low GS activity, under different light intensities (200, 500 and 1000 µmol m−2 s−1), leaf temperatures (27·5, 32·5 and 37·5 °C) and atmospheric O2 concentrations ([O2]: 2, 21 and 40 %, corresponding to 20, 210 and 400 mmol mol−1).
Key Results
An increase in [O2] increased AER in the two cultivars, accompanied by a decrease in PG due to enhanced photorespiration, but did not greatly influence Tr and gS. There were significant positive correlations between AER and photorespiration in both cultivars. Increasing light intensity increased AER, PG, Tr and gS in both cultivars, whereas increasing leaf temperature increased AER and Tr but slightly decreased PG and gS. ‘Kasalath’ (low GS activity) showed higher AER than ‘Akenohoshi’ (high GS activity) at high light intensity, leaf temperature and [O2].
Our results demonstrate that photorespiration is strongly involved in NH3 emission by rice leaves and suggest that differences in AER between cultivars result from their different GS activities, which would result in different capacities for reassimilation of photorespiratory NH3. The results also suggest that NH3 emission in rice leaves is not directly controlled by transpiration and stomatal conductance.
PMCID: PMC3197464  PMID: 21937483
Ammonia assimilation; ammonia emission; glutamine synthetase; nitrogen; Oryza sativa; photorespiration; rice cultivars
2.  Structural and biochemical characterization of the C3–C4 intermediate Brassica gravinae and relatives, with particular reference to cellular distribution of Rubisco 
Journal of Experimental Botany  2011;62(15):5347-5355.
On the basis of its CO2 compensation concentration, Brassica gravinae Ten. has been reported to be a C3–C4 intermediate. This study investigated the structural and biochemical features of photosynthetic metabolism in B. gravinae. The cellular distribution of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) was also examined in B. gravinae, B. napus L. (C3), Raphanus sativus L. (C3), and Diplotaxis tenuifolia (L.) DC. (C3–C4) by immunogold electron microscopy to elucidate Rubisco expression during the evolution from C3 to C3–C4 intermediate plants. The bundle sheath (BS) cells of B. gravinae contained centrifugally located chloroplasts as well as centripetally located chloroplasts and mitochondria. Glycine decarboxylase P-protein was localized in the BS mitochondria. Brassica gravinae had low C4 enzyme activities and high activities of Rubisco and photorespiratory enzymes, suggesting that it reduces photorespiratory CO2 loss by the glycine shuttle. In B. gravinae, the labelling density of Rubisco was higher in the mesophyll chloroplasts than in the BS chloroplasts. A similar cellular pattern was found in other Brassicaceae species. These data demonstrate that, during the evolution from C3 to C3–C4 intermediate plants, the intercellular pattern of Rubisco expression did not change greatly, although the amount of chloroplasts in the BS cells increased. It also appears that intracellular variation in Rubisco distribution may occur within the BS cells of B. gravinae.
PMCID: PMC3223036  PMID: 21825284
Brassica gravinae; Brassicaceae; bundle sheath cell; C3–C4 intermediate plant; glycine decarboxylase; leaf anatomy; photorespiration; ribulose 1,5-bisphosphate carboxylase/oxygenase
3.  Ectopic Overexpression of The Transcription Factor OsGLK1 Induces Chloroplast Development in Non-Green Rice Cells 
Plant and Cell Physiology  2009;50(11):1933-1949.
For systematic and genome-wide analyses of rice gene functions, we took advantage of the full-length cDNA overexpresser (FOX) gene-hunting system and generated >12 000 independent FOX-rice lines from >25 000 rice calli treated with the rice-FOX Agrobacterium library. We found two FOX-rice lines generating green calli on a callus-inducing medium containing 2,4-D, on which wild-type rice calli became ivory yellow. In both lines, OsGLK1 cDNA encoding a GARP transcription factor was ectopically overexpressed. Using rice expression-microarray and northern blot analyses, we found that a large number of nucleus-encoded genes involved in chloroplast functions were highly expressed and transcripts of plastid-encoded genes, psaA, psbA and rbcL, increased in the OsGLK1-FOX calli. Transmission electron microscopy showed the existence of differentiated chloroplasts with grana stacks in OsGLK1-FOX calli cells. However, in darkness, OsGLK1-FOX calli did not show a green color or develop grana stacks. Furthermore, we found developed chloroplasts in vascular bundle and bundle sheath cells of coleoptiles and leaves from OsGLK1-FOX seedlings. The OsGLK1-FOX calli exhibited high photosynthetic activity and were able to grow on sucrose-depleted media, indicating that developed chloroplasts in OsGLK1-FOX rice calli are functional and active. We also observed that the endogenous OsGLK1 mRNA level increased synchronously with the greening of wild-type calli after transfer to plantlet regeneration medium. These results strongly suggest that OsGLK1 regulates chloroplast development under the control of light and phytohormones, and that it is a key regulator of chloroplast development.
PMCID: PMC2775961  PMID: 19808806
Chloroplast development • FOX hunting system • GARP transcription factor • OsGLK1 • Oryza sativa • Rice
4.  Leaf Vascular Systems in C3 and C4 Grasses: A Two-dimensional Analysis 
Annals of Botany  2006;97(4):611-621.
• Background and Aims It is well documented that C4 grasses have a shorter distance between longitudinal veins in the leaves than C3 grasses. In grass leaves, however, veins with different structures and functions are differentiated: large longitudinal veins, small longitudinal veins and transverse veins. Thus, the densities of the three types of vein in leaves of C3 and C4 grasses were investigated from a two-dimensional perspective.
• Methods Vein densities in cleared leaves of 15 C3 and 26 C4 grasses representing different taxonomic groups and photosynthetic subtypes were analysed.
• Key Results The C4 grasses had denser transverse veins and denser small longitudinal veins than the C3 grasses (1·9 and 2·1 times in interveinal distance), but there was no significant difference in large longitudinal veins. The total length of the three vein types per unit area in the C4 grasses was 2·1 times that in the C3 grasses. The ratio of transverse vein length to total vein length was 14·3 % in C3 grasses and 9·9 % in C4 grasses. The C3 grasses generally had greater species variation in the vascular distances than the C4 grasses. The bambusoid and panicoid C3 grasses tended to have a denser vascular system than the festucoid C3 grasses. There were no significant differences in the interveinal distances of the three vein types between C4 subtypes, although the NADP-malic enzyme grasses tended to have a shorter distance between small longitudinal veins than the NAD-malic enzyme and phosphoenolpyruvate carboxykinase grasses.
• Conclusions It seems that C4 grasses have structurally a superior photosynthate translocation and water distribution system by developing denser networks of small longitudinal and transverse veins, while keeping a constant density of large longitudinal veins. The bambusoid and panicoid C3 grasses have a vascular system that is more similar to that in C4 grasses than to that in the festucoid C3 grasses.
PMCID: PMC2803656  PMID: 16464879
C3 and C4 photosynthesis; interveinal distance; longitudinal vein; photosynthetic type; Poaceae; transverse vein
5.  Variation in the Activity of Some Enzymes of Photorespiratory Metabolism in C4 Grasses 
Annals of Botany  2005;96(5):863-869.
• Background and Aims Photorespiration occurs in C4 plants, although rates are small compared with C3 plants. The amount of glycine decarboxylase in the bundle sheath (BS) varies among C4 grasses and is positively correlated with the granal index (ratio of the length of appressed thylakoid membranes to the total length of all thylakoid membranes) of the BS chloroplasts: C4 grasses with high granal index contained more glycine decarboxylase per unit leaf area than those with low granal index, probably reflecting the differences in O2 production from photosystem II and the potential photorespiratory capacity. Thus, it is hypothesized that the activities of peroxisomal enzymes involved in photorespiration are also correlated with the granal development.
• Methods The granal development in BS chloroplasts was investigated and activities of the photorespiratory enzymes assayed in 28 C4 grasses and seven C3 grasses.
• Key Results The NADP–malic enzyme grasses were divided into two groups: one with low granal index and the other with relatively high granal index in the BS chloroplasts. Both the NAD–malic enzyme and phosphoenolpyruvate carboxykinase grasses had high granal index in the BS chloroplasts. No statistically significant differences were found in activity of hydroxypyruvate reductase between the C3 and C4 grasses, or between the C4 subtypes. The activity of glycolate oxidase and catalase were smaller in the C4 grasses than in the C3 grasses. Among the C4 subtypes, glycolate oxidase activities were significantly smaller in the NADP–malic enzyme grasses with low granal index in the BS chloroplasts, compared with in the C4 grasses with substantial grana in the BS chloroplasts.
• Conclusions There is interspecies variation in glycolate oxidase activity associated with the granal development in the BS chloroplasts and the O2 production from photosystem II, which suggests different potential photorespiration capacities among C4 grasses.
PMCID: PMC4247052  PMID: 16100226
Bundle sheath cell; catalase; C4 grasses; C4 subtypes; glycolate oxidase; granal development; hydroxypyruvate reductase; photorespiration; photorespiratory enzymes

Results 1-5 (5)