缺乏叶绿素的油菜突变体的叶绿体组成和结构变化(英文)

对黄化突变体Cr352 9和野生型油菜 (BrassicanapusL .) 352 9叶绿体的超微结构和组成进行了比较。与野生型相比 ,突变体Cr352 9叶片具有较少的类囊体、较少的垛叠膜区和较少的叶绿素含量。突变体的Chla/Chlb比值较高 ,是野生型的 2倍。电泳结果表明 ,突变体类囊体膜中LHCⅡ和其三聚体LHCⅡ 的含量减少。SDS_PAGE分析显示 ,LHCⅡ的脱辅基蛋白在突变体类囊体膜中明显减少。免疫印迹进一步表明 ,所有LHCⅡ组分的含量仅为野生油菜的类囊体膜的 1 / 3。突变体Cr352 9的天线系统比野生型 352 9的小。     

缺乏叶绿素的油菜突变休的叶绿体组成和结构变化

对黄化突变体Cr3529和野生型油菜（Brassica napus L.)3529叶绿体的超微结构和组成进行了比较,与野生型相比,突变体Cr3529叶片具有较少的类囊体,较少的垛叠膜区和较少的叶绿素含量,突变体的Chla/Chob比值较高,是野生型的2倍,电泳结果表明,突变体类囊体膜中LHCII和其三聚体LHCII的含量减少,SDS－PAGE分析显示,LHCII的脱辅基蛋白在突变体类囊体中明显减少,免疫印迹进一步表明,所有LHCII组分的含量仅为野生油菜的类囊体膜的1／3,突变体Cr3529的天线系统比野生型3529的小。     

野生和黄化大麦类囊体膜色素蛋白的分离和比较

用混和去垢剂（ＳＤＳ、ＯＧ）增溶和温和电泳方法,对野生和黄化大麦的叶绿体类囊体膜进行了色素蛋白复合体组成的分离和比较,并结合ＳＤＳ－聚丙烯酰胺凝胶电泳对类囊体膜蛋白进行了鉴定。结果表明,黄化大麦类囊体膜色素蛋白组分中,ＬＨＣ Ⅱ和ＬＨＣ Ⅰ的含量都较野生型大麦明显减少;并讨论了黄化大麦类囊体膜色素蛋白含量的减少与其光合速率降低的关系。     

黄化油菜突变体Cr3529子叶类囊体膜光谱性质研究

以发育10d的黄化油菜突变体为材料,分析了突变体油菜子叶类囊体膜的色素含量、室温吸收光谱、叶绿素荧光发射和激发光谱以及蛋白内源荧光光谱的变化。数据显示：与野生型相比,突变体油菜子叶类囊体膜的光合色素Chl α和Chl b含量均减少．但Chl α／b比值升高;突变体油菜子叶类囊体膜叶绿素捕光能力和受激发能力均下降,且较依赖于Chl α捕光并将光能激发传递给PSⅡ反应中心;突变体油菜子叶类囊体膜的蛋白内源荧光也明显异于野生型。进一步表明突变体油菜子叶类囊体膜蛋白组成发生了改变。     

黄瓜叶片光合电子传递对水分胁迫的响应

黄瓜叶片在水分胁迫下叶片相对含水量减少,类囊体室温吸收光谱的吸收峰降低,同时其NADP光还原活性、Ca^2 -ATPase活性也相应降低,全链电子传递明显受阻。类囊体膜蛋白电泳分析结果显示：类囊体膜色素蛋白复合体含量有不同程度的降低,其中PSⅡ色素蛋白复合体含量下降较多,试验结果表明水分胁迫通过限制光能的吸收,传递双及转换效率,抑制了光合电子传递过程。     

低叶绿素b高产水稻突变体及其光合特性的研究

最近发现了一个在田间条件下自然产生的低叶绿素ｂ高产水稻突变体（Ｏｒｙｚａ ｓａｔｉｖａ Ｌ．ｃｖ．Ｚｈｅｎｈｕｉ ２４９）,该突变体主要降低了外周捕光天线复合本的含量。这种变化主要表现在叶片全展前后,到叶片发育后期则接近野生型。与以往所研究的突变体不同的是,该突变体叶绿素ｂ含量仅适量减少,因而不影响类囊体膜的稳定性。突变体的光合机构在叶片一生中较稳定,这可能表明突变减少了光系统截获的光能,相对提高     

低叶绿素b高产水稻突变体及其光合特性的研究

最近发现了一个在田间条件下自然产生的低叶绿素b高产水稻突变体(Oryza sativa L. cv.Zhenhui 249),该突变体主要降低了外周捕光天线复合体的含量.这种变化主要表现在叶片全展前后,到叶片发育后期则接近野生型.与以往所研究的突变体不同的是,该突变体叶绿素b含量仅适量减少,因而不影响类囊体膜的稳定性.突变体的光合机构在叶片一生中较稳定,这可能表明突变减少了光系统截获的光能,相对提高了光能的利用率,减少了O-2的产生.     

盐酸胍处理大麦类囊体膜对金属阳离子调节激发能分配的影响

以大麦（ＨｏｒｄｅｕｍＶｕｌｇａｒｅＬ）叶片为实验材料,研究盐酸胍处理大麦类囊体膜对金属阳离子调节激发能分配的影响。结果表明：当未用盐酸胍处理时,Ｍｇ２＋对类囊体膜的Ｆ６８６／Ｆ７３６比值有显著的刺激作用,而用不同浓度盐酸胍处理类囊体膜,则明显阻滞Ｍｇ２＋调节激发能向ＰＳＩＩ分配,表明盐酸胍处理可能损伤类囊膜上ＬＨＣ—ＩＩ与Ｍｇ２＋调节作用有关的蛋白质,从而导致Ｍｇ２＋调节效应降低．用ＳＤＳ—ＰＡＧＥ方法研究盐酸胍处理后的类囊体膜色素蛋白复合物的组成变化,结果表明盐酸胍处理导致ＭＩＣ—ＩＩ中ＭＩＣＰ１和ＬＨＣＰ２组分消失,这说明盐酸胍处理阻滞Ｍｇ２＋调节效应可能与ＬＪＩＣ—ＩＩ上的ＬＨＣＰ１,ＬＨＣＰ２受损有关。     

水稻叶绿体基粒缺乏突变体的叶绿素蛋白质复合体分析

我们曾报道一个细胞核隐性基因控制的黄绿色水稻突变体。其叶片的叶绿素含量仅为正常品种叶片的1/3,Chla:Chlb比值很高;叶绿体缺乏基粒,即基粒数和每一基粒的片层数均很少;但以叶绿素量为基数的光合活性较高。为了认识色素含量、基粒结构和光合功能之间相互关系,对正常品种和突变体叶绿体膜的叶绿素蛋白质复合体作了比较分析。结果表明,突变体缺乏24KD的捕光叶绿素a/b蛋白质复合体,但富有28KD的叶绿素a蛋白质复合体CPa。从试验结果推论,叶绿体片层的垛叠和捕光叶绿素a/b蛋白质复合体的含量有正的相关,非垛叠片层则富有叶绿素a蛋白质复合体CPa;     

叶片阶段性白化小麦的叶绿体激发能分配和荧光动力学特征

叶片阶段性白化小麦是一种非常特殊的色素突变体。本文对其白色、白绿相嵌及转绿叶片的叶绿体光化活性分别进行了测定。试验表明：（１）白化和白绿相嵌叶片的叶绿体色素合成处于停滞阶段,转绿叶片则处于色素迅速合成阶段;（２）白化、白绿相嵌和转绿叶片叶绿体的激发能传递呈递增趋势;（３）三者对绿体的ＰＳⅡ活性和原初光能转化效率也呈递增趋势;（４）白化叶片叶绿体缺乏ＰＳⅠ和ＰＳⅡ外周天线色素蛋白,白绿相嵌和转绿叶片的类囊体膜多肽组分与正常叶片相近似。     

Suppression subtractive hybridization identifies differentially expressed genes in Brassica napus chlorophyll-reduced mutant

Suppressive subtraction hybridization (SSH) was used to identify differentially expressed genes caused by a chlorophyll-reduced mutation in B. napus. The cDNA fragments, derived from SSH positive subtractive library (tester: normal wild type, driver: mutant) were cloned into pMD18-T vector. Two hundred SSH cDNA clones were screened by dot blot array, and 151 clones were identified as differentially expressed cDNA fragments in Cr3529 line. Thirty-six positive clones which showed marked expression differences were selected and sequenced. After redundant cDNAs were removed, 33 differentially expressed unique cDNA section clones were obtained. Among the 33 clones, two clones possess different parts of the cDNA sequence of the same gene coding geranylgeranyl reductase, four clones belong to unknown proteins, and the rest share homology to genes of diverse class. Sequence analysis showed that at least 12 genes were discovered to be related to the photosynthesis, seven of them coded the proteins which belong to the subunit of photosystem 2. RNA gel blot analysis showed that compared with 3529, the gene expression of the chlorophyll a/b-binding protein Lhcb2 in photosystem 2 declined markedly in the cotyledons and seedling leaves of Cr3529, indicating that the reduced light-harvesting complex 2 accumulation in thylakoid membrane of Cr3529 was due to the decrease of the related gene mRNA level for translation.     

Developmental Loss of Photosystem II Activity and Structure in a Chloroplast-Encoded Tobacco Mutant, Lutescens-1

Lutescens-1, a tobacco mutant with a maternally inherited dysfunction, displayed an unusual developmental phenotype. In vivo measurement of chlorophyll fluorescence revealed deterioration in photosystem II (PSII) function as leaves expanded. Analysis of thylakoid membrane proteins by polyacrylamide gel electrophoresis indicated the physical loss of nuclear- and chloroplast-encoded polypeptides comprising the PSII core complex concomitant with loss of activity. Freeze fracture electron micrographs of mutant thylakoids showed a reduced density, compared to wild type, of the EF_s particles which have been shown previously to be the structural entity containing PSII core complexes and associated pigment-proteins. The selective loss of PSII cores from thylakoids resulted in a higher ratio of antenna chlorophyll to reaction centers and an altered 77 K chlorophyll fluorescence emission spectra; these data are interpreted to indicate functional isolation of light-harvesting chlorophyll a/b complexes in the absence of PSII centers. Examination of PSII reaction centers (which were present at lower levels in mutant membranes) by monitoring the light-dependent phosphorylation of PSII polypeptides and flash-induced O₂ evolution patterns demonstrated that the PSII cores which were assembled in mutant thylakoids were functionally identical to those of wild type. We conclude that the lutescens-1 mutation affected the correct stoichiometry of PSII centers, in relation to other membrane constituents, by disrupting the proper assembly and maintenance of PSII complexes in lutescens-1 thylakoid membranes.     

Identification Mutants of Plant Pigment with Spectral Technology

Differences in the pigment and thylakoid membranes levels among the mutant barley 1832C, Chlorina-f2 was compared with a normal one by means of their absorption spectra and the 4th derivative spectra. Results showed that there was no absorption peaks for Chl b at 651 nm in red and 470 nm in blue regions in the absorption and the 4th derivative spectra in the mutant thylakoid membranes. At the same time there was no absorption peaks of Chl b at 645 nm and 455 nm in the absorption and the 4th derivative spectra in its acetone (80%) extract. Therefore, mutant barley 1832C was proved to be a new type of chlorophyll b-less mutant that could survive independently in nature.     

用光谱技术鉴定植物色素突变体

1983年我国报道了从γ-射线处理的“矮杆齐”大麦中得到了一株黄绿色的突变体1832C[1]。本文用光谱技术对该突变体的光合色素成分进行了鉴定。1　材料和方法　　材料为六棱裸大麦“矮杆齐”和由该品种大麦诱变形成的黄绿色突变体1832C(Mb1832C),以及作为对照的缺失Chlb的突变体大麦Chlorina-f2[2]都于3月初播种于实验田中。　　每个样品取30g新鲜的叶片,先用自来水后用蒸馏水冲洗干净。把洗净的叶片摊放在干净的纱布上吸干表面水分,剪碎,加入100mL预冷的含有0.4mol/L山梨醇、0.1mol/LTris-HCl(pH7.6)的缓冲液,用组织捣碎机先慢速捣碎1…     

A new chlorophyll-protein complex related to Photosystem I in Chlamydomonas reinhardii

A new chlorophyll-protein complex, CP O, was isolated from Chlamydomonas reinhardii using lithium dodecyl sulfate polyacrylamide gel electrophoresis run at 4°C. A similar complex is recovered using Triton/digitonin solubilization of thylakoid membranes of the F54-14 mutant lacking in CP I and ATPase. CP O is enriched in long-wavelength chlorophyll a and contains five polypeptides (27.5, 27, 25, 23 and 19 kDa). Its 77 K fluorescence emission spectrum peaks at 705 nm while CP II have an emission maximum at 682 and 720 nm, respectively. Comparison of the polypeptide pattern of the wild type and AC40 mutant of C. reinhardii shows that the five CP O polypeptides are specifically lacking in the mutant. Although the 77 K emission originating from the Photosystem (PS) I pigments is lower in the mutant than in the wild type, the two spectra show the same peaks at 686, 694 and 717 nm. However, comparison of the 77 K emission spectrum of the F14 mutant lacking in CP I with that of the double mutant AC40-14 lacking in CP I and CP O shows the absence in the latter of the large emission band peaking at 707 nm. The 707 nm emission is thought to arise from some PS I antennae and is quenched in the wild type by the presence of PS I traps located in CP I. We conclude that CP O is a part of the PS I antenna in C. reinhardii which controls the 707 nm fluorescence emission.     

Vipp1 is required for basic thylakoid membrane formation but not for the assembly of thylakoid protein complexes.

Vipp1 (vesicle inducing protein in plastids 1) is found in cyanobacteria and chloroplasts where it is essential for thylakoid formation. Arabidopsis thaliana mutant plants with a reduction of Vipp1 to about 20% of wild type content become albinotic at an early stage. We propose that this drastic phenotype results from an inability of the remaining Vipp1 protein to assemble into a homo-oligomeric complex, indicating that oligomerization is a prerequisite for Vipp1 function. A Vipp1-ProteinA fusion protein, expressed in the Deltavipp1 mutant background, is able to reinstate oligomerization and restore photoautotrophic growth. Plants containing Vipp1-ProteinA in amounts comparable to Vipp1 in the wild type exhibit a wild type phenotype. However, plants with a reduced amount of Vipp1-ProteinA protein are growth-retarded and significantly paler than the wild type. This phenotype is caused by a decrease in thylakoid membrane content and a concomitant reduction in photosynthetic activity. To the extent that thylakoid membranes are made in these plants they are properly assembled with protein-pigment complexes and are photosynthetically active. This strongly supports a function of Vipp1 in basic thylakoid membrane formation and not in the functional assembly of thylakoid protein complexes. Intriguingly, electron microscopic analysis shows that chloroplasts in the mutant plants are not equally affected by the Vipp1 shortage. Indeed, a wide range of different stages of thylakoid development ranging from wild-type-like chloroplasts to plastids nearly devoid of thylakoids can be observed in organelles of one and the same cell.     

Altered chloroplast structure and function in a mutant of Arabidopsis deficient in plastid glycerol-3-phosphate acyltransferase activity

Mutants of Arabidopsis thaliana deficient in plastid glycerol-3-phosphate acyltransferase activity have altered chloroplast membrane lipid composition. This caused an increase in the number of regions of appressed membrane per chloroplast and a decrease in the average number of thylakoid membranes in the appressed regions. The net effect was a significant decrease in the ratio of appressed to nonappressed membranes. A comparison of 77 K fluorescence emission spectra of thylakoid membranes from the mutant and wild type indicated that the ultrastructural changes were associated with an altered distribution of excitation energy transfer from antenna chlorophyll to photosystem II and photosystem I in the mutant. The changes in leaf lipid composition did not significantly affect growth or development of the mutant under standard conditions. However, at temperatures above 28°C the mutant grew slightly more rapidly than the wild type, and measurements of temperature-induced fluorescence yield enhancement suggested an increased thermal stability of the photosynthetic apparatus of the mutant. These effects are consistent with other evidence suggesting that membrane lipid composition is an important determinant of chloroplast structure but has relatively minor direct effects on the function of the membrane proteins associated with photosynthetic electron transport.     

Absence of lutein, violaxanthin and neoxanthin affects the functional chlorophyll antenna size of photosystem-II but not that of photosystem-I in the green alga Chlamydomonas reinhardtii

Chlamydomonas reinhardtii double mutant npq2 lor1 lacks the beta, epsilon-carotenoids lutein and loroxanthin as well as all beta,beta-epoxycarotenoids derived from zeaxanthin (e.g. violaxanthin and neoxanthin). Thus, the only carotenoids present in the thylakoid membranes of the npq2 lor1 cells are beta-carotene and zeaxanthin. The effect of these mutations on the photochemical apparatus assembly and function was investigated. In cells of the mutant strain, the content of photosystem-II (PSII) and photosystem-I (PSI) was similar to that of the wild type, but npq2 lor1 had a significantly smaller PSII light-harvesting Chl antenna size. In contrast, the Chl antenna size of PSI was not truncated in the mutant. SDS-PAGE and Western blot analysis qualitatively revealed the presence of all LHCII and LHCI apoproteins in the thylakoid membrane of the mutant. The results showed that some of the LHCII and most of the LHCI were assembled and functionally connected with PSII and PSI, respectively. Photon conversion efficiency measurements, based on the initial slope of the light-saturation curve of photosynthesis and on the yield of Chl a fluorescence in vivo, showed similar efficiencies. However, a significantly greater light intensity was required for the saturation of photosynthesis in the mutant than in the wild type. It is concluded that zeaxanthin can successfully replace lutein and violaxanthin in most of the functional light-harvesting antenna of the npq2 lor1 mutant.     

Spectral properties of a divinyl chlorophyll a harboring mutant of Synechocystis sp. PCC6803

A divinyl chlorophyll (DV-Chl) a harboring mutant of Synechocystis sp. PCC 6803, in which chlorophyll species is replaced from monovinyl(normal)-Chl a to DV-Chl a, was characterized. The efficiency of light utilization for photosynthesis was decreased in the mutant. Absorption spectra at 77 K and their fourth derivative analyses revealed that peaks of each chlorophyll forms were blue-shifted by 1–2 nm, suggesting lowered stability of chlorophylls at their binding sites. This was also true both in PSI and PSII complexes. On the other hand, fluorescence emission spectra measured at 77 K were not different between wild type and the mutant. This indicates that the mode of interaction between chlorophyll and its binding pockets responsible for emitting fluorescence at 77 K is not altered in the mutant. P700 difference spectra of thylakoid membranes and PSI complexes showed that the spectrum in Soret region was red-shifted by 7 nm in the mutant. This is a characteristic feature of DV-Chl a. Microenvironments of iron–sulfur center of a terminal electron acceptor of PSI complex, P430, were practically the same as that of wild type.