条浒苔蛋白核超微结构和Rubisco及其活化酶分子定位

研究了条浒苔蛋白核超微结构及其Rubisco和Rubisco活化酶金相免疫分子定位。超微结构显示,条浒苔细胞具有形态及组成相同的1～2个蛋白核,每个蛋白核被淀粉鞘所包围。蛋白核中央均有1条由1个类囊体构成的纵向孔道,并有时局部特别膨大。纵向孔道两端与叶绿体基质相连接。小球藻Rubisco抗体对条浒苔Rubisco的Western印迹图谱显示仅为1条带,其位置与SDS-PAGE电泳图谱上的主带相对应,分子量大约为55kD。金相免疫分子定位的结果显示,条浒苔Rubisco金标颗粒主要分布于叶绿体的蛋白核(71.86%)和淀粉鞘(27.94%)部位中,按面积密度计算二者总和占99.8%,极少分布在叶绿体类囊体和基质中(0.2%)。Rubisco活化酶分子定位也显示其主要分布于蛋白核和淀粉鞘中。这些结果均表明条浒苔蛋白核(及淀粉鞘)与单细胞绿藻的蛋白核相同,具有光合作用功能。条浒苔Rubisco初始活性和总活性的测定结果表明,其活化率较高,高达77.62%。     

Responses of leaf photosynthesis and plant growth to altered source–sink balance in a Japanese and a Dutch tomato cultivar

We investigated the effects of altered source/sink ratio by leaf or fruit pruning on leaf photosynthetic characteristics and whole-plant growth of ‘Momotaro York’, a Japanese cultivar, and ‘Dundee’, a Dutch cultivar and verified a hypothesis for sink-limitation of plant growth proposed by Tanaka and Fujita (1974). Plants were grown hydroponically with a high-wire system in a greenhouse for 11 weeks. Light-saturated photosynthesis of young, fully expanded leaves measured at atmospheric CO₂ partial pressure of 37 Pa and at an intercellular CO₂ partial pressure of 20 Pa was not influenced by alteration of source/sink ratio for either cultivar. Although soluble sugars were accumulated in leaves under high source/sink conditions, the amount of ribulose-1,5-bisphosphate carboxylase/oxygenase, a rate-limiting factor for CO₂-limited photosynthesis, was not affected by the altered source–sink balance. Net assimilation rate of a whole plant increased with decreasing source/sink ratio, but this can be accounted for by the changes in light interception per unit leaf area, without taking the source–sink relationship into consideration. It was concluded that the altered source/sink ratio did not change leaf photosynthetic capacity and the sink-limitation hypothesis cannot be applied to either cultivar under the conditions of the present study.     

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.     

Mechanism of High Photosynthetic Capacity in BC2F4 LinesDerived from a Cross between Oryza sativa and Wild Relatives O. rufipogon

Abstract

We found that several BC₂F₄ lines had high leaf photosynthetic rates under light-saturated and ambient CO₂ conditions. These lines are progenies of BC₂F₁ plants with high photosynthetic capacities which were generated by backcrossing between Oryza rufipogon (W630) and O. sativa cv. Nipponbare, as a recurrent parent. Some photosynthetic characteristics of the BC₂F₄ lines were investigated to identify the factors increasing photosynthetic rates. Photosynthetic rates of these lines under light-saturated conditions at 50 to 700 ppm CO₂ concentrations were higher than those in Nipponbare. The estimated-maximum photosynthetic rates under light-saturated and CO₂-saturated conditions in BC₂F₄ lines were also higher than that in Nipponbare. The photosynthetic rate under light-saturated and ambient CO₂ conditions was positively correlated with the carboxylation efficiency as an indicator of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco) activity in vivo rather than stomatal conductance. Initial and total Rubisco activities in vitro tended to be higher in the BC₂F₄ lines than in Nipponbare. The content of active Rubisco calculated from the activation state of Rubisco was also higher in the BC₂F₄ lines than in Nipponbare. These results suggest that high photosynthetic capacities of BC₂F₁ plants can be maintained high in their progenies and high photosynthetic rates under light-saturated and ambient CO₂ conditions in the BC₂F₄ lines are achieved mainly by the high activity of Rubisco due to the high active Rubisco content.     

Quantifying key model parameters for wheat leaf gas exchange under different environmental conditions

The maximum carboxylation rate of Rubisco(V_(cmax)) and maximum rate of electron transport(J_(max)) for the biochemical photosynthetic model, and the slope(m) of the Ball-Berry stomatal conductance model influence gas exchange estimates between plants and the atmosphere. However, there is limited data on the variation of these three parameters for annual crops under different environmental conditions. Gas exchange measurements of light and CO_2 response curves on leaves of winter wheat and spring wheat were conducted during the wheat growing season under different environmental conditions. There were no significant differences for V_(cmax), J_(max) or m between the two wheat types. The seasonal variation of V_(cmax), J_(max) and m for spring wheat was not pronounced, except a rapid decrease for V_(cmax) and J_(max) at the end of growing season. V_(cmax) and J_(max) show no significant changes during soil drying until light saturated stomatal conductance(gssat) was smaller than 0.15 mol m~(–2) s~(–1). Meanwhile, there was a significant difference in m during two different water supply conditions separated by gssat at 0.15 mol m~(–2) s~(–1). Furthermore, the misestimation of V_(cmax) and J_(max) had great impacts on the net photosynthesis rate simulation, whereas, the underestimation of m resulted in underestimated stomatal conductance and transpiration rate and an overestimation of water use efficiency. Our work demonstrates that the impact of severe environmental conditions and specific growing stages on the variation of key model parameters should be taken into account for simulating gas exchange between plants and the atmosphere. Meanwhile, modification of m and V_(cmax)(and J_(max)) successively based on water stress severity might be adopted to simulate gas exchange between plants and the atmosphere under drought.     

Effects of temperature on growth and photosynthesis in the seedling stage of the sheath blight-resistant rice genotype 32R

The 32R rice genotype is resistant to sheath blight disease (ShB), with a high-yield potential. We examined effects of temperature on the plant responses of 32R in comparison with those of the ShB-susceptible rice genotype (29S) and Nipponbare (Nb, a Japonica standard cultivar). The seedlings at the 4th leaf stage of rice genotypes were exposed to 14/14, 19/14, 25/20, 31/26, 37/32 and 37/37 °C (day/night) for 5, 10 and 15 days. The dry weight, leaf area, photosynthesis, contents of ribulose 1,5-bisphosphate carboxylase/oxygenase (Rubisco) and chlorophyll contents were examined. The dry weight showed lower in 32R than in 29S and Nb at a low temperature, and total dry weight correlated strongly with root dry weight and leaf area. The relative growth rate (RGR) correlated strongly with the net assimilation rate (NAR). Rubisco, chlorophyll contents and the photosynthetic rates were limited at a low temperature and showed lower in 32R than in 29S and Nb. The strong correlations between Rubisco and the rates of maximum photosynthesis and initial slope were found in 32R, but not found in 29S and Nb. In addition, RGR and NAR of 32R correlated positively with Rubisco. These suggest that 32R contains traits of cold-sensitive genotypes that are related to limiting Rubisco at a low temperature, thus diminishing photosynthesis and limiting plant growth. Differences of growth among 32R, 29S and Nb were discussed in the relation of genotypes.     

小麦旗叶Rubisco和Rubisco活化酶与光合作用日变化的关系

Rubisco是植物光合作用中的关键酶,其体内活性由Rubisco活化酶调节.研究了小麦旗叶的光合速率与Rubisco和Rubisco活化酶之间的相关性.结果表明,小麦旗叶的光合速率和Rubisco初始活力、Rubisco活化酶活力都存在着明显的日变化;小麦旗叶的光合速率主要取决于Rubisco的初始活力,即Rubisco被Rubisco活化酶活化的程度,而与Rubisco和Rubisco活化酶本身的含量关系不大.     

自然休眠期低温处理时间对生长期草莓叶片光合特性的影响

以草莓为料材,研究自然休眠期不同低温处理时间对其生长期叶片光合特性的影响.结果表明:在自然休眠过程中,随着低温处理时间延长,草莓新生叶叶面积、叶柄长度不断增加,而初生叶柄角度不断变小,完全通过休眠后,不再有显著变化.新生叶片的光合速率、气孔导度、叶绿素质量比和光合作用关键酶活性也随着低温处理时间延长而不断升高,当低温处理时间大于765 h后,不再有显著变化.自然休眠过程中,低温处理时间可通过调节气孔因素、叶绿素质量比和光合作用关键酶活性来调控生长期草莓叶片的光合作用,当完全通过休眠后,低温处理时间对草莓的生长发育和叶片的光合特性不再有显著影响.     

黄瓜幼苗Rubisco与Rubisco活化酶对光强的响应

以津优3号幼苗为试材,研究弱光(LL:100 μmol· m-2·s-1)、亚弱光(SL:300 μmol·m-2·s-1)下黄瓜幼苗光合速率(Pn)、核酮糖-1,5-二磷酸羧化/加氧酶(Rubisco)、Rubisco活化酶(RCA)活性及其基因表达量的变化.结果表明,11d弱光处理后,黄瓜幼苗叶片的Pn、Rubis...     

小麦叶片胞间隙蛋白的提取与鉴定

采用真空渗入离心法,通过特定的装置并在不同的离心力和时间条件下分离小麦叶片胞间隙液,结合十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)技术,以胞内高含量的蛋白-Rubisco大亚基为胞内物污染的指示,利用高效价的兔抗Rubisco抗体进行Western-blotting检测分析。结果表明,在离心力为3 000g,离心时间为10,15和20 min所提的胞间隙液量最大,且均检测到Rubisco的污染。结合胞间隙液的产率确定小麦叶片胞间隙液提取最佳的离心条件应为离心力1 000~2 500 g,时间15~20 min。