Kurilosides A1, A2, C1, D,E and F—Triterpene Glycosides from the Far Eastern Sea Cucumber Thyonidium (= Duasmodactyla) kurilensis (Levin): Structures with Unusual Non-Holostane Aglycones and Cytotoxicities

Six new monosulfated triterpene tetra-, penta- and hexaosides, namely, the kurilosides A₁ (1), A₂ (2), C₁ (3), D (4), E (5) and F (6), as well as the known earlier kuriloside A (7), having unusual non-holostane aglycones without lactone, have been isolated from the sea cucumber Thyonidium (= Duasmodactyla) kurilensis (Levin) (Cucumariidae, Dendrochirotida), collected in the Sea of Okhotsk near Onekotan Island from a depth of 100 m. Structures of the glycosides were established by 2D NMR spectroscopy and HR-ESI mass spectrometry. Kurilosides of the groups A and E contain carbohydrate moieties with a rare architecture (a pentasaccharide branched by C(4) Xyl1), differing from each other in the second monosaccharide residue (quinovose or glucose, correspondingly); kurilosides of the group C are characterized by a unique tetrasaccharide branched by a C(4) Xyl1 sugar chain; and kurilosides of the groups D and F are hexaosides differing from each other in the presence of an O-methyl group in the fourth (terminal) sugar unit. All these glycosides contain a sulfate group at C-6 of the glucose residue attached to C-4 Xyl1 and the non-holostane aglycones have a 9(11) double bond and lack γ-lactone. The cytotoxic activities of compounds 1–7 against mouse neuroblastoma Neuro 2a, normal epithelial JB-6 cells and erythrocytes were studied. Kuriloside A₁ (1) was the most active compound in the series, demonstrating strong cytotoxicity against the erythrocytes and JB-6 cells and a moderate effect against Neuro 2a cells.     

Ultrastructure of Fusion Product between Protoplasts from C3 and C4 Species of Amaranthaceae

Summary

This paper describes the ultrastructure of the electric field-induced fusion products of C₃ and C₄ species of Amaranthaceae at the early developmental stage. Protoplasts of C₃ species were isolated from a Ceiosia cristata L. cell suspension and, those of C₄ species were isolated from an Amaranthus tricolor L. cotyledon. Incompatibility occurred in the C₃/C₄ hybrid. The incompatibility reactions were detected in the newly formed hybrid cells accompanied with significant changes in the nucleolus (segregation of nucleolar components) and plastids (cup-like shape or amoeboid plastid enclosing cytoplasmic materials) of C₃ species parent. The structural changes in the organelles of the C₄ partner were less marked. After 5 days of culture, most organelles showed high cellular activity, and a normal dedifferentiation process of mesophyll chloroplasts was observed. At this stage nucleolar segregation was not detected and the C₃ species plastids were difficult to distinguish from the proplastids formed from mesophyll chloroplasts. In addition, some mitochondria showed bursting-like structure. However, under the culture condition used these somatic incompatibility did not seem to impair further growth of fusion products since they were still proliferating well resulting in callus formation.     

Photosynthetic Characteristics of an Amphibious C4 Plant,Eleocharis retroflexa ssp. chaetaria

Abstract

Eleocharis retroflexa (Poir.) Urban ssp. chaetaria (Roem. & Schult.) T. Koyama, an amphibious leafless sedge, grows not only under terrestrial conditions but also under completely submerged aquatic conditions. We investigated the photosynthetic traits and structural features of the culms, which are the photosynthetic organs, in the terrestrial and submerged forms of this species and compared them with those of other amphibious species of Eleocharis which are known to change the photosynthetic modes. The culms of the terrestrial form had Kranz anatomy with well-developed Kranz (bundle sheath) cells and high levels of C₄ enzyme activity typical of the NAD-malic enzyme (NAD-ME) subtype of C₄ metabolism. They also had a δ ¹³C value typical of C₄ plants, indicating that the terrestrial form fixes carbon through the C₄ pathway. The culms of the submerged form had not only a Kranz-like anatomy but also revealed anatomical traits typical of leaves of submerged aquatic plants. The activities of the C₄ enzymes in the submerged form were lower than those in the terrestrial form, but were still in the range typical of G4 plants, ¹⁴C pulse-¹²C chase experiments with the submerged form indicated that almost all of the fixed ¹⁴C was incorporated into G4 compounds, and subsequently the raioactivity was transferred into C₃ compounds and sucrose. The submerged form showed no diurnal fluctuation in malate level. These data demonstrate that a C₄ metabolism is operative even in the submerged form. This unique amphibious C₄ plant provides an intriguing example of the physiological and ecological adaptability of C₄plants.     

Unusual Structures and Cytotoxicities of Chitonoidosides A,A1, B,C, D,and E,Six Triterpene Glycosides from the Far Eastern Sea Cucumber Psolus chitonoides

Six new triterpene tetra-, penta- and hexaosides, chitonoidosides A (1), A₁ (2), B (3), C (4), D (5), and E (6), containing one or two sulfate groups, have been isolated from the Far-Eastern sea cucumber Psolus chitonoides, collected near Bering Island (Commander Islands) from the depth of 100–150 m. Three of the isolated compounds (1, 3 and 6) are characterized by the unusual aglycone of new type having 18(20)-ether bond and lacking a lactone in contrast with wide spread holostane derivatives. Another unexpected finding is 3-O-methylxylose residue as a terminal unit in the carbohydrate chains of chitonoidosides B (3), C (4), and E (6), which has never been found before in the glycosides from holothurians belonging to the Psolidae family. Moreover, this monosaccharide is sulfated in the compound 4 into unprecedented 3-O-methylxylose 4-O-sulfate residue. Chitonoidoside C (4) is characterized by tetrasaccharide moiety lacking a part of the bottom semi-chain, but having disaccharide fragment attached to C-4 of Xyl1. Such architecture is not common in sea cucumber glycosides. Cytotoxic activities of the compounds 1–5 against mouse and human erythrocytes and human cancer cell lines: adenocarcinoma HeLa, colorectal adenocarcinoma DLD-1, and leukemia promyeloblast HL-60 cells were studied. The cytotoxic effect of chitonoidoside d (5) was the most significant in this series due to the presence of pentasaccharide disulfated sugar chain in combination with holostane aglycone. Surprisingly, the glycosides 1 and 3, comprising the new aglycone without γ-lactone, demonstrated similar activity to the known compounds with holostane aglycones. Chitonoidoside C (4) was less cytotoxic due to the different architecture of the carbohydrate chain compared to the other glycosides and probably due to the presence of a sulfate group at C-4 in 3-O-MeXyl4.     

Chondrogenic differentiation of rat mesenchymal stem cells on silk fibroin/chondroitin sulfate/hyaluronic acid ternary scaffolds

Cartilage repair is a challenge in bone tissue reconstruction. In this study, silk fibroin (SF), chondroitin sulfate (CS) and hyaluronic acid (HA) were employed to fabricate scaffolds for tissue engineered cartilage by freeze drying technique. The secondary pores were formed in the main pores of SF/CS/HA scaffold which improved the pore connectivity and equilibrium swelling of the scaffold. Furthermore, rat bone marrow mesenchymal stem cells were seeded on the scaffolds to evaluate the cell adhesion and proliferation. Results of hematoxylin/eosin staining and cell counting kit-8 assay showed that the cells migration and differentiation of SF/CS/HA (80/15/5) scaffold were better than that of SF/CS/HA scaffolds with different ratios after 7 days culture. Moreover, immunohistochemistry and scanning electron microscope demonstrated that large amounts of collagen II and proteoglycans of the cells were expressed in the SF/CS/HA 3D scaffold, while the expression of collagen I was barely visible by immunohistochemistry. Abound of extracellular matrix was formed to morphologically round and distributed uniformly throughout the scaffolds. The 3D ternary scaffold could promote the cells chondrogenic differentiation without using any inductive agent and offer potential for cartilage tissue regeneration.     

Effects of partial root-zone drying on yield,tuber size and water use efficiency in potato under field conditions

Water resources are limited for irrigation worldwide; therefore, there is a need for water-saving irrigation practices to be explored. Partial root-zone drying (PRD) is a new water-saving irrigation strategy being tested in many crop species. Experiments were conducted in potato (Solanum tuberosum L. cv. Folva) under open field conditions in 2004 and under a mobile rainout shelter in 2005. Two subsurface irrigation treatments were studied: full irrigation (FI) receiving 100% of evaporative demands, 50.1 and 201 mm of irrigation water in the 2 years, to keep it close to field capacity; and PRD, which received 21.7 and 140 mm of irrigation in 2004 and 2005 respectively. Due to rain in 2004, the PRD treatment was imposed over a short period only during the late tuber filling and maturing stages. In 2005, the PRD treatment was imposed during the whole period of tuber filling and tuber maturation. The PRD treatment was shifted from one side to the other side of potato plants every 5–10 days. Especially in 2005 it was apparent that stomatal conductance was generally lower in the PRD than in the FI plants, whereas leaf water potential tended to be lower in only a few instances. During the treatment period, plants were harvested five times, and no significant difference was found between the treatments in leaf area index, top dry mass and tuber yield. At final harvest, tubers were graded based on size into four classes C₁–C₄, of which the yield of the important marketable class (C₂) was significantly higher (20%) in the PRD than in the FI treatment. Compared with FI, the PRD treatment saved 30% of irrigation water while maintaining tuber yield, leading to a 61% increase of irrigation water use efficiency. The limited data of 2004 support these results. In summary, PRD is a promising water-saving irrigation strategy for potato production in areas with limited water resources.     

Variegatusides: New Non-Sulphated Triterpene Glycosides from the Sea Cucumber Stichopus variegates Semper

Four new triterpene glycosides, variegatusides C–F (1–4), together with three structurally known triterpene glycosides, variegatusides A and B (5, 6), and holothurin B (7), were isolated from the sea cucumber Stichopus variegates Semper (Holothuriidae), collected from the South China Sea. Their structures were elucidated on the basis of extensive spectral analysis (nuclear magnetic resonance (NMR) and electrospray ionization mass spectrometry (ESIMS)) and chemical evidence. Variegatusides C–F exhibit the same structural feature consisting of the presence of a 23-hydroxyl group at the holostane-type triterpene aglycone side chain. Variegatuside C (1) has a double bond (24, 25) in this same chain, while variegatuside D (2) exhibits a 8(9)-ene bond in the holostane-type triterpene aglycone, which has not been extracted from other sea cucumber species. Compound 4 is a native compound from the sea cucumber S. variegates Semper, which has been reported to be desacetylstichloroside B₁. Except for holothurin B, these glycosides have no sulfate group in their sugar chain and show potent antifungal activities in vitro biotests.     

Alkylresorcinol composition allows the differentiation of Triticum spp. having different degrees of ploidy

Total alkylresorcinol (AR) content and homologue composition were assessed in whole grain flours of 15 varieties each of bread wheat, durum, spelt, emmer, and einkorn grown in four different environments. Bread wheat (761 ± 92 μg/g DM) and spelt (743 ± 57 μg/g) belonging to the hexaploid species showed higher AR concentrations than the tetraploid durum (654 ± 48 μg/g, p < 0.05), while the concentrations found in the diploid einkorn (737 ± 91 μg/g) and the tetraploid emmer (697 ± 94 μg/g) did not significantly differ from the other species. The AR content showed a remarkable heritability and, thus, seemed to be mainly determined by genetic factors. If ARs were assumed to be deposited within a specific AR-rich layer of the kernel, AR levels of all varieties would easily surpass their minimal inhibitory concentrations against fungal pathogens within this barrier layer. Although the AR carrying a C21:0 side chain was the main homologue in all species, the levels of all AR homologues and their relative composition significantly differed between hexaploid (bread wheat and spelt), tetraploid (durum and emmer) and diploid (einkorn) species. Consequently, a clear-cut differentiation of Triticum species and derived whole grain flours according to their degrees of ploidy was established based on concentrations of saturated C₁₇-, C₁₉-, C₂₁-, C₂₃-, and C₂₅-substituted ARs.     

More lightweight and isodiametric seeds for C4 than for C3 grasses are associated with preference for open habitats of C4 grasses in a temperate flora

Seed mass and shape of grasses were analysed in a temperate flora containing 178 and 26 species belonging to C₃ and C₄ photosynthetic types respectively. The weedy character and the annual or perennial status were also considered. On the basis of the seed traits studied, three groups were distinguished: C₄ grasses, annual C₃ grasses of weedy character and perennial non‐weedy C₃ grasses. The C₄ group had more isodiametric (same diameter in all directions) seed shape and lower average seed mass than the C₃ group. To our knowledge, this has not yet been described for temperate C₄ grasses and is certainly associated with their preference for open habitats where competition for light is small. Weedy annual C₃ grasses had heavier and less isodiametric seeds than C₄ grasses did. These species are mostly specialized to establish in the dense cover of perennial vegetation, and this ability distinguishes them from the C₄ group. Non‐weedy perennial C₃ species possessed less isodiametric seeds than did C₄ grasses, but did not differ from weedy annual C₃ grasses. As most alien C₄ grasses naturalized in Hungary are annuals with small, isodiametric seeds, these traits are good candidates to be included in screening for potential future invasives in open habitats.     

Ultrastructure of Hybrid Callus between C3 and C4 Species of Amaranthaceae

Summary

Unlike the parent line, the hybrid calli between C₃ and C₄ species of Amaranthaceae have poor division capability. Therefore, in this study the fine structures of hybrid callus derived from protoplasts of Celosia cristata L. cv. Pink Charm (C₃ species) cell suspension and Amaranthus tricolor L. cv. Perfecta (C₄ species) cotyledon callus were investigated by light and electron microscopy, and compared with the parental. All callus lines were composed of parenchymatous cells possessing a voluminous vacuole. Parental callus lines contained organelles with a relatively normal structure. The peculiar feature characterizing the C₃/C₄ hybrid callus was the presence of highly lobed nucleus with multinucleoli and numerous small vacuoles having autophagic activity scattered in the cytoplasm. The autophagic activity is apparently related to the existence of several inclusions such as cytoplasmic organelles in the central vacuole. It might also be related to the pronounced expansion of central vacuole and the reduced cytoplasm. The failure to sustain proliferation related to some atypical features of the organelles is discussed.     

Examining the impact of shade on above-ground biomass and normalized difference vegetation index of C3 and C4 grass species in North-Western NSW,Australia

Previous investigations have detected a directional trend in the normalized difference vegetation index (NDVI) of pastures around scattered paddock trees and identified shade from the tree as the most likely causal factor. This study uses a field experiment to quantify the effect of varying levels of shade on the above-ground biomass and NDVI of three grass species native to Australia (Microlaena stipoides, C₃, shade tolerant; Austrodanthonia richardsonii, C₃, prefers full sunlight, and Chloris ventricosa, C₄, prefers full sunlight) in different seasons. The study demonstrates that shade had little influence on the above-ground biomass of C₃ species but significantly reduced biomass in the C₄ species. Until early winter, the NDVI of each species was generally significantly higher in all shaded treatments than in the no-shade treatment. This suggests that shaded plants retained a higher proportion of green biomass and/or changed leaf shape, increased leaf area and chlorophyll content. Regardless, although not proven in this experiment, it is likely shade prolonged the retention of green plant material into mid to late winter. Overall, this experiment explains the directional trends in NDVI around scattered trees found in previous work and suggests that shade from scattered trees prolongs green pasture production in a range of native grass species, without loss of C₃ pasture biomass.     

小麦近缘染色体异附加系的磷效率及其机理研究

磷素缺乏是限制小麦高产的重要原因,而小麦近缘野生种往往表现出更好的耐低磷胁迫特性。本研究以通过盆栽实验初步筛选出来的9个磷高效的外缘染色体异附加系小麦为供试材料,以它们的亲本中国春(CS)为对照,在水培条件下对异附加系小麦在苗期的磷效率(包括磷吸收和磷利用效率)以及与磷效率有关的生理机制进行了研究。结果表明:(1)有6个外缘染色体异附加系材料(0226、0232、0244、0278、0282、0291)在水培条件下表现出了高磷效率;(2)在这6份磷高效的外缘染色体异附加系材料中有两份材料(0226和0232)在低磷条件下酸性磷酸酶的活性以及草酸和琥珀酸的分泌量都显著高于其亲本CS,而且在低磷条件下这两份材料的Cmin显著低于CS,说明这两份材料中添加的外缘染色体中包含与磷高效有关的基因,因此这两份外缘染色体异附加系可作为小麦育种材料,进而选育出高产的磷高效小麦品种。     

Recombinant Expression and Characterization of α-Conotoxin LvIA in Escherichia coli

α-Conotoxin LvIA is derived from Conus lividus, native to Hainan, and is the most selective inhibitor of α3β2 nicotinic acetylcholine receptors (nAChRs) known to date. In this study, an efficient approach for the production of recombinant α-Conotoxin LvIA is described. Tandem repeats of a LvIA gene fragment were constructed and fused with a KSI gene and a His₆ tag in a Escherichia coli (E. coli) expression vector pET-31b(+). The recombinant plasmids were transformed into E. coli and were found to express well. The KSI-(LvIA)_n-His₆ fusion protein was purified by metal affinity chromatography and then cleaved with CNBr to release recombinant LvIA (rLvIA). High yields of fusion protein ranging from 100 to 500 mg/L culture were obtained. The pharmacological profile of rLvIA was determined by two-electrode voltage-clamp electrophysiology in Xenopus laevis oocytes expressing rat nAChR subtypes. The rLvIA antagonized the α3β2 nAChR subtype selectively with a nano-molar IC₅₀. The rLvIA was analgesic in a mouse hot-plate test model of pain. Overall, this study provides an effective method to synthesize α-conotoxin LvIA in an E. coli recombinant expression system, and this approach could be useful to obtain active conopeptides in large quantity and at low cost.     

Spectroscopic and chromatographic evaluation of the wax ester fraction of Adenanthera pavonina oil

Present study was carried out in order to assess the constituents of Adenanthera pavonina wax esters by Fourier Transform Infrared (FT-IR) spectroscopy and Gas chromatography (GC). After extraction of oil from A. pavonina seeds, wax esters were separated from oil by centrifugation and then purified through crystallization process in acetone. The white crystalline solid with melting point of 83-85 °C was initially characterized as wax esters by FT-IR. Gas chromatography with flame ionization detector (GC-FID) provided the separation of various wax esters from C₃₄ to C₄₈ including odd and even chain esters. C₄₆ wax ester was found to be predominant. For separation, identification and estimation of individual constituents of fatty acid methyl esters and fatty alcohol acetates were carried out by GC-MS after saponification and derivatization of wax esters. GC-MS analysis indicated the presence of saturated methyl esters from C_l4 to C₃₀. Predominant methyl ester was C₂₂. Wax esters contained the saturated fatty alcohols from C₁₄ to C₂₈. The major alcohol was saturated C₂₄ fatty alcohol. The results indicated that A. pavonina wax esters have potential to be used in pharmaceutical and cosmetic industry due to its better composition of long chain fatty acids and fatty alcohols.     

Influence de la fumure azotée sous diverses conditions de culture sur les constituants non-sucrés azotés et sur la qualité technologique de la betterave à sucre

Samples of sugar beet from a fertilizer experiment testing five doses of (NH₄)₂SO₄ (0–2000 kg/ha), were analysed in order to determine the influence of different levels of nitrogen fertilizer on the undesirable nitrogen components in beet-roots. The chemical analyses were carried out at the end of winter (February-March), when all moisture in the soil was due to precipitation only; and in summer (May-June), when the effect of five soil moisture levels (irrigation treatments), in addition to fertilizer levels, was examined. A linear, highly significant increase in the following nitrogen components of the roots was found with increasing fertilizer doses: total N, aminoacid N and N amide +NH₃. With time there was a decrease in the above nitrogen components, which was significant in half the cases. Contrary to the finding concerning fertilizer effects, no correlation could be established linking irrigation treatments to absolute N component values in the roots. However, relative values (No=100%) showed that prolonging irrigation to the end of May reduced the negative influence of nitrogen fertilizer on the quality of roots. Present work shows that a fertilizer dressing exceeding 1200 kg/ha ammonium sulfate may reduce beet quality for processing, causes technological difficulties during the manufacturing process, and loss of sugar.     

Preparation of cellulose/multi-walled carbon nanotube composite membranes with enhanced conductive property regulated by ionic liquids

Cellulose/multi-walled carbon nanotubes (MWCNTs)- composite membranes applied in electrochemical and biomedical fields were prepared using 1-ethyl-3-methylimidazolium diethyl phosphate (EmimDEP) as solvent in this study. With the increasing of MWCNTs amount, the membrane conductivity increased, and the conductivity reached 9.1 S/cm as the mass ratio of MWCNTs to cellulose being 2:1. The additions of sodium dodecyl sulfate (SDS), 1-hexadecyl-3-methylimidazolium bromide (C₁₆mimBr) and 1-butyl-3-methylimidazolium tetrafluoroborate (BmimBF₄) efficiently improved the conductivity, mechanical property, and thermal stability by promoting the dispersion of MWCNTs. When the mass ratio of C₁₆mimBr to MWCNTs changed from 0 to 0.3:1, the conductivity increased from 0.08 S/cm to 0.14 S/cm, and the tensile strength increased from 13.3 MPa to 17.0 MPa. These results indicate that the binary ionic liquids (ILs) system can regulate the properties of the composite membranes, and is a feasible approach for preparing cellulose/MWCNTs composite membranes with enhanced properties.     

Presence of chloroplasts in mestome sheath cells of the C3 Pooid grass Elymus tsukushiensis

ABSTRACT

The successful introduction of the C₄ pathway into C₃ crops would increase photosynthetic rates and crop productivity. However, our poor understanding of how Kranz leaf anatomy develops poses a great obstacle. In particular, the origin, development, and genetics of bundle sheath (BS) cells in C₄ plants are key points to elucidate. Here we report that Elymus tsukushiensis, a common C₃ grass of the subfamily Pooideae, contains chloroplasts in the mestome sheath (MS) cells of the leaf, unlike most MS cells of C₃ grasses. The chloroplasts are smaller than those of mesophyll cells. Immunogold localization showed that the chloroplasts and mitochondria of MS cells, respectively, accumulate ribulose 1,5-bisphosphate carboxylase/oxygenase and a photorespiratory enzyme, glycine decarboxylase, as in mesophyll cells. Thus, we suggest that the MS cells have weak photosynthetic and photorespiratory functions. This finding provides an insight into the development and evolution of C₄-type BS cells in leaves of C₃ grasses.     

Comprehensive utilization of the hydrolyzed productions from rice hull

The study based on pretreatment, hydrolyzation and separation processes with the raw material rice hull, provides a comprehensive utilization of the hydrolyzed productions, such as glucose (C₆H₁₂O₆) from cellulose, silica (SiO₂), and byproduct crystalline sodium sulfate (Na₂SO₄·10H₂O). The optimum hydrolysis conditions are as follows: the concentration of H₂SO₄ is 72% (wt.%), the temperature is 50 °C, the ratio of H₂SO₄ solution volume (mL) to the rice hull mass (g) is 10:1 and the time is 5 min, the glucose yield rate reaches 45.6% (wt.%). The concentration of glucose solution could be 0.1 g/mL after neutralization measured by ultraviolet spectrophotometer (UV-VIS). Silica powder was below 50 nm characterized by X-ray diffraction (XRD) and scanning electron microscopy (SEM). The main byproduct crystalline sodium sulfate was featured by XRD and photographs.     

A study of methods of incorporation and source of14C on distribution of radioactivity in potato plants

Russet Burbank potato plants grown in the greenhouse were given¹⁴C as¹⁴CO₂ and as uniformly labelled sucrose. Labelled sucrose was applied; (1) by injection into the main stem, (2) injection with a hypodermic needle into the debladed petiole and (3) uptake of labelled sucrose kept in a vial fastened to the main stem using thread as a wick. Plants harvested 6 hours later were analyzed for radioactivity in the foliage, roots, and tubers. The greatest proportion of radioactivity was found in the foliage. Tubers were analyzed for radioactivity in the basic (amino acids), acidic (organic acids and sugar phosphates), neutral (sugars) and alcohol insoluble solid (starch) fractions. Most of the radioactivity was found in the sugar component of the chemical fraction. Plants treated with¹⁴⁷CO₂ had a higher percentage of radioactivity in the acidic and starch fractions than plants treated with sucrose. Sucrose applied by petiole injection not only showed higher total incorporation but also exhibited more translocation of labelled material into the tubers than the other methods.     

Rubisco small subunits of C4 plants,Napier grass and guinea grass confer C4-like catalytic properties on Rubisco in rice

ABSTRACT

Overexpression of Rubisco small subunit (RbcS) of C₄ plant, sorghum (sorghum bicolor) was shown to enhance the catalytic turnover rate (k _cat) of Rubisco in rice (Oryza sativa). In this study, the effects of other Rubisco small subunits of C₄ plants, Napier grass (Pennisetum purpureum) and guinea grass (Megathyrsus maximus) on kinetic properties of Rubisco in rice were studied. The expression levels of Napier grass RbcS (NgRbcS) and guinea grass RbcS (GgRbcS) proteins accounted for 41% and 45% of total RbcS, respectively in homozygous overexpression lines. The k _cat and K _m for CO₂ (Kc) of Rubisco were increased in all transgenic lines. Interestingly, the k _cat was markedly higher in NgRbcS homozygous line, whereas K _c was notably higher in GgRbcS homozygous line. Although its effects depend on species, these results suggest that the introduction of C₄ RbcS are effective approaches to alter the catalytic properties of Rubisco in rice.