水稻叶绿素合成缺陷突变体及其生物学研究进展

叶绿素是植物叶绿体内参与光合作用的重要色素,叶绿素合成缺陷突变是一类明显的性状突变,其在理论研究和实际应用方面均具有重要的意义。本文介绍了国内外在水稻叶绿素合成缺陷突变体的发掘、作用机理及其基因定位等方面的研究进展。     

蒺藜苜蓿糖基转移酶基因SMALL AND EMERALD1的克隆和功能研究

类黄酮代谢对于植物生长发育和植物-环境互作至关重要,其中糖基转移酶介导的糖基化修饰在类黄酮代谢中发挥着重要作用。为了研究蒺藜苜蓿中糖基转移酶的生物学功能,通过定向筛选蒺藜苜蓿Tnt1逆转座子插入突变体库,获得了一类植株矮小、叶片深绿的突变体small and emerald1 (se1)。通过基因表型连锁性分析成功克隆了SE1基因,该基因编码1个糖基转移酶,与拟南芥中调控类黄酮生物合成的AtUGT84A1氨基酸同源性为52.8%。对野生型和se1突变体叶片的类黄酮含量进行测定发现类黄酮总量在se1突变体中显著降低(P<0.01)。进一步研究发现在se1突变体中类黄酮合成途径关键基因CHS、F3H和F3’H表达水平下降。亚细胞定位显示SE1可能在细胞质和细胞核中发挥生物学功能。研究表明糖基转移酶基因SE1可能参与蒺藜苜蓿类黄酮合成代谢调控,进而影响其生长发育。此外,研究还发现SE1基因对于叶绿素合成可能具有负向调控作用。     

花色素苷对拟南芥耐盐性的影响

为探讨花色素苷在盐胁迫中的防御作用及其机制,以模式植物拟南芥(Arabidopsis thaliana)花色素苷合成途径相关基因缺失突变体(DFR基因缺失突变体tt3,CHS基因缺失突变体tt4,CHS、DFR基因双缺失突变体tt3tt4)及其野生型(WT)为材料,采用叶绿素荧光和超氧阴离子(O_2·~–)组织定位等方法,分析了tt3,tt4,tt3tt4和WT对盐胁迫处理的生理响应。结果表明,盐胁迫下3种缺失突变体叶片花色素苷含量的增加显著低于野生型,与WT相比,叶绿素荧光参数Fv/Fm、Yield、ETR、q P和NPQ下降较快,膜渗漏率升高显著,叶片O_2·~–积累程度为tt3tt4tt3/tt4WT。这表明花色素苷在植物抵御盐胁迫过程中起着重要作用,它可能是作为渗透调节剂及抗氧化剂来增强植物的耐盐性。因此,花色素苷含量可以作为筛选耐盐作物的指标。     

水稻黄绿叶突变体研究进展

叶绿素是植物光合作用的重要色素,叶绿素的合成决定植物的光合效率,并且直接影响作物的产量和品质。叶绿素的合成及分解代谢是一个非常复杂的过程,在此过程中有较多的基因参与,其中任何一个基因发生突变都有可能影响叶绿素的合成及分解,从而使叶片表现出各种叶色变化或者会影响植物的生长。叶色突变体研究是探明叶绿体发育过程中基因功能的有效途径。因此,发掘和鉴定叶色突变基因,开展与叶绿素相关基因的定位、克隆及功能研究均具有重要的理论意义和应用价值。综述了水稻黄绿叶突变体的研究进展,旨在为研究水稻叶绿素生物合成途径和光合作用机制提供理想的材料,同时还可作为标记性状在杂种优势中进行利用。     

文心兰浅绿条纹突变体的生理生化及叶绿素荧光特性研究

以文心兰浅绿条纹突变体为材料,分析叶片光合色素含量和组成、叶绿素合成前体物质含量以及叶绿素荧光参数的变化,观察突变体叶绿体超微结构的改变,以探寻其叶色变异的生理基础。结果表明:(1)突变体叶绿素a(Chl a)、叶绿素b(Chl b)、类胡萝卜素(Car)和总叶绿素(Chl)含量分别比叶色正常植株显著降低了37.1%、34.0%、30.8%和36.3%。(2)突变体叶绿素生物合成受阻于胆色素原(PBG)到尿卟啉原Ⅲ(UrogenⅢ)的反应步骤。(3)突变体叶绿体发育存在明显的缺陷,基粒数目及基粒片层的垛叠层数明显减少,嗜锇颗粒及囊泡较多。(4)突变体初始荧光(Fo)比正常植株高39%,最大荧光(Fm)、最大光化学效率(Fv/Fm)、PSⅡ有效光化学效率(Fv′/Fm′)和PSⅡ实际光化学效率(ΦPSⅡ)均显著低于正常植株,但光化学淬灭系数(qP)和非光化学淬灭系数(NPQ)与正常植株无显著差异。研究结果说明,文心兰叶绿素生物合成受阻和叶绿体结构发育不良,导致叶绿素的含量下降,致使突变体叶片呈现浅绿条纹,光能利用率降低。     

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。     

小麦突变体返白系返白阶段叶绿素代谢的变化

小麦 ( Triticum aestivum)返白系在返白阶段 Chl、胡萝卜素 ( Caro)含量均下降 ,但 Caro/ Chl的比值大于对照 ,表明叶片白化不是因 Caro减少引起的。Chl下降的同时 ,Chla和 Chlb均下降 ,表明该突变体属阶段性缺总 Chl型。返白初期 Chlase活性增高 ,返白中期活性下降 ,表明 Chl降解不是造成叶片失绿的主要原因 ;Chl合成中间物 δ-氨基酮戊酸 ( ALA)、胆色素原 ( PBG)积累 ,尿卟啉 ( Uro )、原卟啉 ( Proto )、镁 -卟啉 ( Mg- Proto )、原叶绿素酸 Pchl减少 ,特别是 Uro 在返白中期含量最低 ,复绿初期却急剧积累 ,表明叶绿素合成受阻于尿卟啉原 ( Urogen )的形成上。     

植物叶色白化研究进展

植物叶色白化是一类明显和常见的叶绿素缺失突变,在植物光合作用机理、激素生理、核一质基因组相互作用、遗传育种等理论研究和实际应用方面均具有无可替代的价值。主要综述了近年来国内外有关白化突变体与叶绿素合成的关系、影响白化的内外因素、相关基因的克隆以及克服白化的途径等方面的研究进展,旨在为深入研究植物叶色白化突变提供参考。     

小麦黄化突变体光合作用及叶绿素荧光特性研究

对小麦自然黄化突变体及其突变亲本(西农1718)的叶绿素含量、光合速率及叶绿素荧光动力学参数进行比较分析.结果显示:(1)突变体金黄株、绿黄株、黄绿株的叶绿素含量均显著低于突变亲本,总叶绿素含量分别为突变亲本的17%、24%和58%,表明该突变体为叶绿素缺乏突变体;3个突变体叶绿素a与叶绿素b的比值(Chl a/Chl b)均小于突变亲本,而且突变体叶绿素含量越低,Chl a/Chl b比值越小,说明该突变体Chl a下降幅度大于Chl b.(2)金黄株净光合速率在孕穗期、开花期仅为突变亲本的5.7%、2.4%;绿黄株净光合速率显著低于突变亲本,为突变亲本的57.7%、43.3%;而叶绿素含量仅为突变亲本一半的黄绿株,其净光合速率接近突变亲本,表明该黄化突变体叶绿素含量在一定范围内单位叶绿素含量的光合效率较高.(3)突变体Fo均显著低于突变亲本;金黄株、绿黄株的Fm,Fv,qP,qN显著低于突变亲本;金黄株Fv/Fm比值(0.671)显著低于突变亲本.研究表明,叶绿素含量在一定范围内减少,未引起突变体叶绿素荧光动力学参数(Fo除外)显著改变,而当叶绿素含量较大程度减少时,这些荧光参数会急剧降低.     

植物叶绿素缺失突变体分子机制研究进展

植物叶绿素缺失突变体在自然界中广泛存在,是研究叶绿素形成和叶绿体发育等代谢途径的良好材料.该文主要从分子层面上阐述了叶绿素缺失突变体产生的原因,如叶绿素合成受阻、叶绿体光合蛋白合成或输入受阻、叶绿体RNA转录物未被编辑、过量光损伤和卟啉循环各物质之间的相互抑制,并归纳了近年来鉴定出来的一些叶绿素缺失突变基因,简要介绍了叶绿素和叶绿体之间的关系以及叶绿素缺失突变体的应用.     

Recent advances in chlorophyll biosynthesis and breakdown in higher plants

Chlorophyll (Chl) has unique and essential roles in photosynthetic light-harvesting and energy transduction, but its biosynthesis, accumulation and degradation is also associated with chloroplast development, photomorphogenesis and chloroplast-nuclear signaling. Biochemical analyses of the enzymatic steps paved the way to the identification of their encoding genes. Thus, important progress has been made in the recent elucidation of almost all genes involved in Chl biosynthesis and breakdown. In addition, analysis of mutants mainly in Arabidopsis, genetically engineered plants and the application of photo-reactive herbicides contributed to the genetic and regulatory characterization of the formation and breakdown of Chl. This review highlights recent progress in Chl metabolism indicating highly regulated pathways from the synthesis of precursors to Chl and its degradation to intermediates, which are not longer photochemically active.     

Chlorobium Tepidum: Insights into the Structure,Physiology, and Metabolism of a Green Sulfur Bacterium Derived from the Complete Genome Sequence

Green sulfur bacteria are obligate, anaerobic photolithoautotrophs that synthesize unique bacteriochlorophylls (BChls) and a unique light-harvesting antenna structure, the chlorosome. One organism, Chlorobium tepidum, has emerged as a model for this group of bacteria primarily due to its relative ease of cultivation and natural transformability. This review focuses on insights into the physiology and biochemistry of the green sulfur bacteria that have been derived from the recently completed analysis of the 2.15-Mb genome of Chl. tepidum. About 40 mutants of Chl. tepidum have been generated within the last 3 years, most of which have been made based on analyses of the genome. This has allowed a nearly complete elucidation of the biosynthetic pathways for the carotenoids and BChls in Chl. tepidum, which include several novel enzymes specific for BChl c biosynthesis. Facilitating these analyses, both BChl c and carotenoid biosynthesis can be completely eliminated in Chl. tepidum. Based particularly on analyses of mutants lacking chlorosome proteins and BChl c, progress has also been made in understanding the structure and biogenesis of chlorosomes. In silico analyses of the presence and absence of genes encoding components involved in electron transfer reactions and carbon assimilation have additionally revealed some of the potential physiological capabilities, limitations, and peculiarities of Chl. tepidum. Surprisingly, some structural components and biosynthetic pathways associated with photosynthesis and energy metabolism in Chl. tepidum are more similar to those in cyanobacteria and plants than to those in other groups of photosynthetic bacteria.     

Chloroplast biogenesis 92: In situ screening for divinyl chlorophyll(ide) a reductase mutants by spectrofluorometry

Chlorophyll biosynthetic heterogeneity is rooted mainly in parallel divinyl (DV) and monovinyl (MV) biosynthetic routes interconnected by 4-vinyl reductases (4VRs) that convert DV tetrapyrroles to MV tetrapyrroles by conversion of the vinyl group at position 4 of the macrocycle to ethyl. What is not clear at this stage is whether the various 4VR activities are catalyzed by one enzyme of broad specificity or by a family of enzymes encoded by one gene or multiple genes with each enzyme having narrow specificity. Additional research is needed to identify the various regulatory components of 4-vinyl reduction. In this undertaking, Arabidopsis mutants that accumulate DV chlorophyllide a and/or DV chlorophyll [Chl(ide)] a are likely to provide an appropriate resource. Because the Arabidopsis genome has been completely sequenced, the best strategy for identifying 4VR and/or putative regulatory 4VR genes is to screen Arabidopsis Chl mutants for DV Chl(ide) a accumulation. In wild-type Arabidopsis, a DV plant species, only MV chlorophyllide (Chlide) a is detectable. However in Chl mutants lacking 4VR activity, DV Chl(ide) a may accumulate in addition to MV Chl(ide) a. In the current work, an in situ assay of DV Chl(ide) a accumulation, suitable for screening a large number of mutants lacking 4-vinyl Chlide a reductase activity with minimal experimental handling, is described. The assay involves homogenization of the tissues in Tris-HCl:glycerol buffer and the recording of Soret excitation spectra at 77K. DV Chlide a formation is detected by a Soret excitation shoulder at 459 nm over a wide range of DV Chlide a/MV Chl a ratios. The DV Chlide a shoulder became undetectable at DV Chlide a/MV Chl a ratios less than 0.049, that is, at a DV Chlide a content of less than 5%.     

Temperature sensitivity as a general phenomenon in a collection of chlorophyll-deficient mutants of sweetclover (Melilotus alba)

A collection of chlorophyll (Chl)-deficient mutants of sweetclover (Melilotus alba) with defects in eight nuclear loci were grown at 17 or 26° C. Plants grown at either temperature were examined for Chl content, Chla/b ratio, expression of the light-harvesting complex II (LHC-II) apoproteins, and protochlorophyllide (Pchlide) biosynthetic capacity. Except for thech4 mutant, the parental strain and all mutants accumulate more Chl when grown at 26° C than at 17° C. Thech5 mutants, lacking Chl b under any growth condition, and thech12 mutant showed little temperature-dependent phenotypic plasticity, whereas this was a marked phenomenon in the other mutants. Thech10 andch11 mutants demonstrated extreme temperature sensitivity with regard to the production of Chlb and the Chlb-binding LHC-II apoproteins. When excised trifoliolates were supplemented with exogenously supplied -aminolevulinic acid, only thech4 mutant was markedly impaired in the ability to produce Pchlide. These data indicate that temperature-sensitive phenotypic plasticity is a common phenomenon of chlorophyll-deficient mutants and substantiate that only a minority of Chl-deficient mutants is impaired in the biosynthesis of Chl.This research was supported by Grants GM84-CRCR-1-1479 (J.C.O.) and 89-00641 (J.M.) of the United States Department of Agriculture and by National Science Foundation Grant DMB87-03100 (J.M.). This is paper No. 8971, Nebraska Agricultural Research Division.     

Identification of Photosynthetic Mutants of Arabidopsis by Automatic Screening for Altered Effective Quantum Yield of Photosystem 2

Quantification of chlorophyll (Chl) fluorescence is a versatile tool for analysing the photosynthetic performance of plants in a non-intrusive manner. A pulse-amplitude modulated fluorometer was combined with a CNC router for the automated measurement of the effective quantum yield of photosystem 2 (₂) of Arabidopsis thaliana plants. About 90 000 individual plants representing 7 500 lines derived from En-transposon and T-DNA mutagenised Arabidopsis populations were screened for mutants with altered ₂. Forty-eight recessive ₂ mutations were identified of which most exhibit also altered pigmentation and increased photosensitivity. For three ₂ mutants the corresponding mutated genes were identified that code all for chloroplast-located proteins. Comparison of the ₂ mutant screen with other screening methods based on the measurement of Chl fluorescence shows that the ₂ mutants identified are different to mutants identified by high Chl fluorescence. Some ₂ mutants, on the contrary, are common to mutants identified by screens based on non-photochemical quenching.     

The Divergent Roles of STAYGREEN (SGR) Homologs in Chlorophyll Degradation

Degradation of chlorophyll (Chl) by Chl catabolic enzymes (CCEs) causes the loss of green color that typically occurs during senescence of leaves. In addition to CCEs, STAYGREEN1 (SGR1) functions as a key regulator of Chl degradation. Although sgr1 mutants in many plant species exhibit a stay-green phenotype, the biochemical function of the SGR1 protein remains elusive. Many recent studies have examined the physiological and molecular roles of SGR1 and its homologs (SGR2 and SGR-LIKE) in Chl metabolism, finding that these proteins have different roles in different species. In this review, we summarize the recent studies on SGR and discuss the most likely functions of SGR homologs.