OsDXR interacts with OsMORF1 to regulate chloroplast development and the RNA editing of chloroplast genes in rice

Plant chlorophyll biosynthesis and chloroplast development are two complex processes that are regulated by exogenous and endogenous factors. In this study, we identified OsDXR, a gene encoding a reductoisomerase that positively regulates chlorophyll biosynthesis and chloroplast development in rice. OsDXR knock-out lines displayed the albino phenotype and could not complete the whole life cycle process. OsDXR was highly expressed in rice leaves, and subcellular localization indicated that OsDXR i...     

Correlation between Chloroplast Ultrastructure and Chlorophyll Fluorescence Characteristics in the Leaves of Rice (Oryza sativa L.) Grown under Salinity

Abstract

The seedlings of Oryza sativa L. cv. Nipponbare grown by hydroponic culture for 3 wks were treated with 75, 100, 150 and 200 mM NaCl for 14, 14, 6 and 3 days, respectively, and examined for chloroplast ultrastructure in the region where chlorophyll fluorescence had been recorded. NaCl treatment decreased the ratio of variable to maximum chlorophyll fluorescence yield (F_v/F_m) and caused swelling of thylakoids. The swelling of thylakoids was quantified by the percentage of the length of swollen thylakoids to the total length of thylakoids. This value was increased with increasing NaCl concentration. Although F_v/F_m decreased at all concentrations of NaCl, the minimal fluorescence yield F₀ was not increased by the treatment with 75 or 100 mM NaCl. The percentage of the length of swelling was low at 75 and 100 mM NaCl. On the other hand, F₀ increased and the swelling of thylakoids was prominent with 150 and 200 mM NaCl treatment. These results suggest that the decrease in F_v/F_m due to the increase in F₀ under salt stress correlates with the ultrastructural damage. The decrease in F_v/F_m due to the increase in F₀ is expected to be useful as an indicator to evaluate the damage in chloroplasts, especially in thylakoid membranes, under salinity.     

巴西橡胶树镁离子转运蛋白基因HbMGT10的克隆及表达分析

Mg~(2+)是植物细胞中含量最高的二价阳离子,是多种酶活性调控的辅因子,在植物的生长发育过程中发挥重要作用;同时,作为叶绿素分子的中心原子,在植物光合作用中起着至关重要的作用,但目前有关高大乔木叶片中Mg~(2+)跨膜运输的研究还很少。MGT/MRS2类型的镁离子转运蛋白在植物镁离子的跨膜运输过程中起着重要的作用,本文克隆研究了一个巴西橡胶树MGT基因,HbMGT10。亚细胞定位显示,HbMGT10定位于叶绿体膜上;酵母互补实验表明,HbMGT10具有镁离子转运功能;qRT-PCR分析结果显示,HbMGT10主要在橡胶树叶片中表达,且是叶片中表达丰度最高的HbMGT基因;HbMGT10在叶片中的表达存在明显的发育调控,随叶片发育进程表达量明显增加,在淡绿期和稳定期表达量最大;HbMGT10在成熟叶片中的表达呈现明显的日变化,在光强度最大的12:00~16:00的表达量最大。由此推测,HbMGT10在橡胶树叶片叶绿体膜的镁离子跨膜转运过程中起着重要的作用,参与了橡胶树叶片叶绿体的发育,以及叶绿体中镁离子浓度的调控,以适应橡胶树叶片的光合作用。     

Evidence of rapid evolution and incipient speciation in Vicia sativa species complex based on nuclear and organellar RFLPs and PCR analysis

Seventy three accessions of the sevenVicia species belonging to Sativa speciescomplex were screened for nuclear and organellar restriction fragmentlength polymorphic (RFLP), and random amplified polymorphicDNA (RAPD) markers. Total genomic DNAs of 73 accessions wasrestricted with three enzymes, and the restriction fragments werehybridized to the wheat rDNA probe pTa71 (containing 18S, 5.8Sand 25S rDNA genes, and spacers), and faba bean probes Ver6-5 (entire intergenic spacer flanked by small part of25S and 18S fragments) and Ver 18-6 (part of thecoding region of the gene and internal transcribed spacers). InXbaI digests, 16 repeat unit length classes in24 combinations were identified. Digestion withEcoRI and DraI gave2–4 and 1–3 fragments, respectively, with detectablehybridization to the probes, indicating the existence of internalXbaI sites. All the accessions produced 3.5 EcoRI fragment arising fromcoding region of the repeat unit. Four hundred and eighteen RAPDmarkers among 45 accessions were identified with 14 arbitrary10-base primers. The percentage of polymorphic bands withinspecies ranged from 20 in V.angustifolia to 98% inV. nigra. Both RFLP andRAPD markers were unable to assess the relationships betweenaccessions within species as there was often much closer resemblancesbetween certain accessions of different species rather than betweenaccessions within each taxon. This analysis supports the view basedon morphological, cytogenetical and crossability data that it is notpossible to classify Sativa species complex into a small finitenumber of taxa which are clearly circumscribed, and that the complexrepresents a unique case of rapid evolution and incipient speciation.A study of chloroplast and mitochondrial RFLPs was undertaken toanalyze phylogeny through maternal lineage. Chloroplast DNArestriction fragment patterns, using 13 restriction endonucleases,revealed 92.6 to 99% homology between the seven species.Twelve enzyme-probe combinations yielded identical fragmentpatterns for all the seven species. The molecular sizes of thechloroplast DNAs obtained were similar (121.5–123.5), indicating that they had all lost one of theinverted repeats. Total DNAs digested with three restriction enzymesand hybridized to six heterologous probes of mitochondrial originyielded monomorphic bands in five enzyme—probe combinationsacross all the 73 accessions. In other combinations as well,40–66 accessions yielded monomorphic profiles. The smallvariation in the remaining accessions was not species-specificsince the same profiles were present in more than one species. Theseresults i) strongly suggest that the seven species within thecomplex share a common ancestor, or direct lineage and, ii)indicate that these species should be relegated to a rank, perhaps ofsubspecies, within V.sativa species complex.     

Phylogeographic study based on intraspecific sequence variation of chloroplast DNA for the conservation of genetic diversity in the Japanese endangered species Primula sieboldii

To gather information on intraspecific phylogeography for use in conservation programs for the endangered species Primula sieboldii in Japan, we analyzed sequence variation in five noncoding regions of chloroplast DNA. Twenty-two distinct haplotypes were recognized in total. The distribution of most haplotypes was geographically confined, but one haplotype was widely distributed throughout northern Japan, and several haplotypes were found in geographically distant regions. Three major clades were revealed by phylogenetic analysis of the haplotypes. Clade I was distributed in Kyushu and central Honshu, clade II in western Honshu and Hokkaido, and clade III from central Honshu to Hokkaido. According to analysis of molecular variance, 59.9% of the total cpDNA variation existed among regions, 32.5% among populations within regions, and 7.6% within populations. Therefore, if genetic conservation of the species is valued, transplanting of P. sieboldii among regions should be avoided. Multiple lineages often existed even in geographically narrow areas (e.g., within a 20-km range), so transplantation between adjacent populations in restoration activities should be carefully designed so as not to change the gene pool of local populations significantly. Also, the geographical distribution of cpDNA haplotypes may allow us to confirm the origin of plants collected for commercial purposes.     

杉木叶绿体发育超微结构的初步研究

<正>自1817年Pelletier和Caventou命名叶绿素以来,人们对叶绿体的结构及其光合作用的研究取得了显著的成就。1865年Sachs报告了叶绿素乃是定位在特殊的小体中,叶绿素只有在光照条件下才能形成,阳光决定了该特殊小体在吸收二氧化碳中的活性。1883年Schimper证实Sachs的特殊小体,并命名为叶绿体。1915年R.M.Willstatter对叶绿素进行了深入的研究。1961年M.Caivin对光合作用进行了研究,揭示了C还原的生化途径,即C_3途径。Miihlethaler和Freg—Wyssling于1959年利用黄化苗进行了叶绿体光合膜形态建成的研究。     

叶色黄化突变体甜瓜叶片内部生理生化变化 的研究

从薄皮甜瓜白莎蜜1 号中筛选出一个生长发育正常的自发型叶色黄化突变体93 88-1,以突
变亲本白莎蜜1 号为对照,对其光合色素含量、叶绿素生物合成前体物质含量、叶绿素荧光参数以及叶绿
体超微结构进行研究。结果显示：9388-1 光合色素含量显著低于白莎蜜1 号,而叶绿素a 与叶绿素b 的
比值显著高于白莎蜜1 号;叶绿素合成受阻于胆色素原（PBG）与尿卟啉原Ⅲ（Urogen Ⅲ）之间;荧光参
数F₀、P_n、ФPS Ⅱ和ETR 显著低于白莎蜜1 号,而qN 和NPQ 显著高于白莎蜜1 号,Fm、FV/Fm 和qP 与
白莎蜜1 号无显著差异;突变体9388-1 的叶绿体内部基粒片层垛叠数有所减少,基粒排列不整齐,呈线
条状,基粒片层间距离大,排列疏松,内含物混浊,淀粉粒少,而白莎蜜1 号基粒片层结构紧密排列,结
构完整,淀粉粒多。表明叶绿素的生物合成受阻,导致叶绿体内部结构发育缺陷,从而影响光合色素的稳
定性,改变叶绿体各种色素的含量与比例,最终引起叶色黄化。同时,突变体能及时地耗散过剩的光能,
对光合机构起一定的保护作用,维持植株正常生长。     

STUDY ON ULTRASTRUCTURE IN SEVERAL TYPICAL HALOPHYTES

<正> The characteristics of form and the structure of typical salt plants are usually consid-ered as suitable to the condition of salinity. So after studying their microstructure, we alsodid more careful observation of their ultrastructure and found some characteristics. Thesecharacteristics can not only explain the way in which they have adapted to salinization but     

The Research of Bt and OC Gene Cotransformation in Tobacco Chloroplast

The Bt Cry IA (C) chloroplast expression cassette and OC chloroplast expression cassette were constructed. The Bt expression cassette contained the 3.5 kb wild type Bt Cry IA (C) gene under the control of the strong light-induced psbA promoter and terminator from rice (Oryza sativa. L) chloroplast, the gene:trnH-psbA-trnk from tobacco (Nicotiana tabacum. L) as the homologous fragment. The OC chloroplast expression cassette contained the OC gene under the control of 16S promoter and terminator from tobacco, the tobacco gene: psbA-ORF512 as homologous fragment. The two cassettes both had the aadA gene expression cassette as the selectable marker. Leaves of tobacco were cotransformed with the particle bombardment method. After selection by spectinomycin, the transformants were obtained. The integration of Bt and OC gene were confirmed by Southern-blotting analysis, and Western-blotting analysis. Proteinase inhibitor assays showed that the Bt and OC gene had expressed. Bioassays showed that the transgenic tobacco had a significant resistance to the larvae of cotton bollworm ( helicoverpa zea ).