水稻致死突变体基因克隆与分子机制研究进展

水稻致死突变体主要有叶片白化致死突变体和叶片早衰致死突变体2大类,这2类突变体相关基因的克隆和功能分析对于解析水稻白化致死和早衰致死具有重要意义,主要涉及光合色素的合成与降解、叶绿体发育与调控、蛋白质的合成与降解、激素合成与调控途径、细胞程序性死亡与转录因子调控等相关基因。本文综述了水稻致死突变基因的克隆与分子机制,并分析了它们的育种价值;并提出在阐明水稻衰老调控机制的基础上,从分子水平和栽培技术上设计调控水稻衰老的策略,发掘其育种和生产价值。

Response of Chlorophyll Fluorescence Characteristic of Trifolium repens L. to Water Stress

[Objective] The aim was to study response of chlorophyll fluorescence parameters in Trifolium repens L.leaves to water stress from perspective of physiology.[Method] With T.repens cultivar ＂Haifa＂ as tested material,three soil water content levels were set to culture plants,including 75% （no stress,CK）,50% （moderate stress,LD） and 25% （severe stress,HD）,the effects of water stress on chlorophyll fluorescence parameters were determined.[Result] The results showed that the chlorophyll fluorescence parameters changed a little when the relative soil moisture content was 75% and 50%,while the chlorophyll fluorescence parameters such as conversion efficiency of primary light energy （Fv/Fm） of PS Ⅱ,the photochemical quenching coefficient （qP） and the quantum yield of PS Ⅱ electron transport （φPS Ⅱ） decreased respectively when the relative soil moisture content was 25%.[Conclusion] When T.repens in severe drought conditions,the physiological functions were damaged,showing symptoms of drought injury.

Photoinhibition and Photooxidation in Leaves of indica and japonica Rice Under Different Temperatures and Light Intensities

Physiological indices related to the efficiency (F-v/F-m) of light energy conversion in PS II and the peroxidation of membrane lipid were measured in leaves of Oryza sativa L. sp. indica rice cv. 'Shanyou 63' and sp. japonica rice cv. '9516'' under different temperatures and fight intensities for 4 days. No changes in F-v/F-m and membrane lipid peroxidation product (MDA) were observed, so neither photoinhibition nor photooxidation happened in both rice cultivars under moderate temperature and medium light intensity. However, F-v/F-m dropped obviously with no change in MDA contents, and photoinhibition appeared in indica rice cv. 'Shanyou 63' under medium temperature and strong light intensity. Furthermore, both photoinhibition and photooxidation were observed in two rice cultivars under chilling temperature and strong light intensity. Experiments with inhibitors under chilling temperature and strong light intensity showed that indica rice had a decrease in DI protein content and SOD activity, and the extent of inhibition of xanthophyll. cycle and nonphotochemical quenching (qN) was larger, and a higher level of MDA was observed. The photoinhibition and photooxidation in indica rice were more distinct as compared with japonica rice. The authors suggested that PS II light energy conversion efficiency (F-v/F-m) and membrane lipid peroxidation were the key indices for the detection of photooxidation.

Effect of TMS(nanostructured silicon dioxide) on growth of Changbai larch seedlings

The roots of 200 one-year-old Changbai Larch (Larix olgensis) seedlings were soaked for 6 hours at the TMS concentrations of 2000, 1000, 500, 250, 125, and 62 μL·L?1. Mean seedling height, root collar diameter, main root length and number of lateral roots were measured every 15 days during growing season from May 30 to Oct. 20. Experimental results showed that TMS treatments greatly promoted seedling growth and improved seedling quality. The treatment by 500 μL·L?1 TMS produced the best result, for which the mean height, root collar diameter, main root length, and the number of lateral roots of seedlings were increased by 42.5%, 30.7%, 14.0%, and 31.6%, respectively, compared to that of the control seedlings. As to seedling quality, grade-I seedling and grade-II seedlings were fifty-fifty, and no grade-III seedlings was found. The treatment by 500 μL·L?1 TMS resulted in the highest chlorophyll concentration. Keywords Changbai Larch - Larix olgensis - Seedling production - Nanostructured silicon dioxide CLC number S143.8 Document code B Foundation item: This study is supported by Jilin Forestry Group Co.Biography: LIN Baoshan (1955-), male, Associate professor at the college of forestry, Beihua University, Jilin City 132011, Jilin Province, P.R China.Responsible editor: Chai Ruihai.

Photochemical Efficiency of PSⅡ and Membrane Lipid Peroxidation in Leaves of indica and japonica Rice (Oryza sativa) Under Chilling Temperature and Strong Light Stress Conditions

Relationships between fluorescence parameters and membrane lipid peroxidation in leaves of indica and japonica rice (Oryza sativa L.) during later growth stage were studied under chilling temperature and strong light stress conditions. Results showed that D1 protein contents of PSⅡ in photosynthetic apparatus dropped, the generation of antheraxanthin (A) and zeaxanthin (Z) of xanthophyll cycle were inhibited partly, PSⅡ photochemical efficiency (F v/F m)and non-photochemical quenching (q N) were also decreased obviously. In addition, endogenous active oxygen scavenger—superoxide dismutase (SOD) reduced, superoxide anion radical (O -· 2) and malondialdehyde (MDA) accumulated, as a result, photooxidation of leaves occurred under chilling temperature and strong light stress conditions. Obvious differences in the changes of the above mentioned physiological parameters between indica and japonica rice were observed. Experiments in leaves treated with inhibitors under chilling temperature and strong light conditions showed that indica rice was more sensitive to chilling temperature with strong light and subjected to photooxidation more than japonica rice. Notable positive correlation between D1 protein contents and F v/F m or (A+Z)/(A+Z+V), and a marked negative correlation between F v/F m and MDA contents were obtained by regression analysis in indica and japonica rice during chilling temperature and strong light conditions. According to the facts mentioned above, it was inferred that PSⅡ photochemical efficiency(F v/F m) was the key index to forecast for the prediction of photooxidation under stress circumstances and the physiological basis were the synthetic capacity of D1 protein and the protection of xanthophyll cycle.

Study on the Degradation Trend of Ryegrass-White Clover Artificial Grassland

[Objective] The study aimed to explore the degradation law and trend of artificial grassland. [Method] Taking the ryegrass （Lolium perenne） - white clover （ Trifolium repens） artificial grassland in Maiping Township, Guizhou Province as the research object, the grassland vegetation of 40 quadrate from different areas （area around the sheep shed, hilltop, hillside, flatland at the foot of the hill） were analyzed by comparing the dominance and richness index. [ Result] Degradation of different degrees appeared in various areas of this artificial grassland. To be specific, the degradation of grassland around the sheep shed was not serious for less consumption by the sheep; however, the areas on the hilltop and hillside degenerated to the grassland types of dallis grass （ Paspalum dilatatum Poir. ） -Indian lovegrass （ Eragrostis pilosa） and dallis grass - cogongrass [ Imperata cylindrica （ Linn. ） Beauv. ], respectively, and the area at the foot of the hill degenerated to the grassland type dominated by garland chrysanthetnum （ Chrysanthemum coronarium L. ） and knotgrass （ Paspalum distichum L. ）. [ Conclusion ] This study provided a basis for grassland improvement as well as the efficient and sustainable utilization of grazing-type artificial grassland in South China.

Photosynthetic Excitation Pressure Causes Violaxanthin De-epoxidation in Aging Cabbage （Brassica Oleracea L.） Leaves

The purpose of the present studies was analysis of the age induced changes in photochemical efficiency and xanthophyils cycle pigments of the primary cabbage （Brassica oleracea L. cv. Capitata f. alba） leaves. Photochemical efficiency of photosystem Ⅱ （PS Ⅱ） was studied by a pulse amplitude modulated chlorophyll fluorescence apparatus, chlorophyll concentration was analysis spectrophotometrically and xanthophyll cycle pigments were estimated by high-pressure liquid chromatography （HPLC）. Leaf senescence was accompanied with a decrease both in chlorophylls concentration, the photochemical efficiency and rate constant for PS Ⅱ photochemistry whereas non-photochemical parameters increased. Excitation pressure （1-qP） which is a measure of relative lumen acidification increased by 1.2x in aging leaves. The maximum quantum yield of PS Ⅱ showed no significant change. The level of de-epoxidised xanthophylls increased but the concentration of mono- and di-epoxy xanthophylls decreased in aging leaves. A linear relationship between the excitation pressure and the depoxidation state of the xanthophyll cycle pigments and lutein, during the onset of senescence suggests that excitation pressure can be used as a sensor for monitoring the onset of senescence as well for the de-epoxidation state of the xanthophylls responsible for non-photochemical quenching in stressed leaves.

EAG and behavioral responses of Helicoverpa armigera males to volatiles from poplar leaves and their combinations with sex pheromone

Electroantennogram (EAG) evaluation of selected compounds from wilted leaves of black poplar, Populus nigra, showed that phenyl acetaldehyde, methyl salicylate, (E)-2-hexenal elicited strong responses from male antennae of Helicoverpa armigera. When mixed with sex pheromone (Ph), some volatiles, e.g. phenyl acetaldehyde, benzyl alcohol, phenylethanol, methylsalicylate, linalool, benzaldehyde, (Z)-3-hexenol, (Z)-3-hexenylacetate, (Z)-6-nonenol, cineole, (E)-2-hexenal, and geraniol elicited stronger responses from male antennae than Ph alone. Wind tunnel bioassay demonstrated that various volatiles could either enhance or inhibit the effect of synthetic sex pheromone. (E)-2-hexenal, (Z)-3-hexenol and linalool in combination with Ph could not induce any male to land on source at all, whereas phenyl acetaldehyde, benzaldehyde, (Z)-6-nonenol and salicylaldehyde combined with Ph enhanced male response rates by 58.63%, 50.33%, 51.85% and 127.78%, respectively, compared to Ph alone. These results suggested that some volatiles shouldmodify sex pheromone caused behavior and that some of them could possibly be used as a tool for disrupting mating or for enhancing the effect of synthetic sex pheromone in the field.

Cellular localization and biochemical characterization of a novel calciumdependent protein kinase from tobacco

By screening tobacco cDNA library with MCK1 as a probe, we isolated a cDNA clone NtCPK5 （accession number AY971376）, which encodes a typical calcium-dependent protein kinase. Sequence analyses indicated that NtCPK5 is related to both CPKs and CRKs superfamilies and has all of the three conserved domains of CPKs. The biochemical activity of NtCPK5 was calcium-dependent. NtCPK5 had Vmax and Km of 526nmol/min/mg and 210μg/ml respectively with calf thymus histone （fraction Ⅲ, abbreviated to histone Ⅲs） as substrate. For substrate syntide-2, NtCPK5 showed a higher Vmax of 2008 nmol/min/mg and a lower Km of 30μM. The K0.5 of calcium activation was 0.04μM or 0.06μM for histone Ⅲs or syntide-2 respectively. The putative myristoylation and palmitoylation consensus sequence of NtCPK5 suggests that it could be a membrane-anchoring protein. Indeed, our transient expression experiments with wild type and mutant forms of NtCPK5/GFP fusion proteins showed that NtCPK5 was localized to the plasma membrane of onion epidermal cells and that the localization required the N-terminal acylation sites of NtCPK5/GFP. Taking together, our data have demonstrated the biochemical characteristics of a novel protein NtCPK5 and its subcellular localization as a membrane-anchoring protein.

Effects of Nitrogen Deposition on Tundra Vegetation Undergoing Invasion by Deyeuxia angustifolia in Changbai Mountains

In recent years, herbaceous species such as Deyeuxia angustifolia （Kom.） Y. L. Chang has invaded alpine tundra regions of the western slope of the Changbai Mountains. Because atmospheric nitrogen deposition is predicted to increase under a warming climate and D. angustifolia is sensitive to nitrogen addition, field experiments were conducted from 2010 to 2013 to determine the effect of increased nitrogen deposition on the mechanisms of D. angustifolia invasion. The goal of this study is to evaluate the impact of increased nitrogen deposition on the changes in alpine tundra vegetation （consisting mostly of Rhododendron chrysanthum Pall. and Vaccinium uliginosum Linn.）. The results showed that： 1） simulated nitrogen deposition affected overall characteristics and structure of R. chrysanthum and E uliginosum communities and had a positive impact on the growth of tundra vegetation invaded by 1）. angustifolia; 2） R. chrysanthum was more resistant to invasion by D. angustifolia than V.. uliginosum; 3） simulated nitrogen deposition could improve the growth and enhance the competitiveness of D. angustifolia, which was gradually replacing R. chrysanthum and V. uliginosum and might become the dominant species in the system in future, transforming alpine tundra into alpine meadow in the Chanebai Mountains.

Disruption of phytoene desaturase gene results in albino and dwarf phenotypes in Arabidopsis by impairing chlorophyll, carotenoid, and gibberellin biosynthesis

Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3） encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification and analysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino and dwarfphenotypes ofthepds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development was arrested at the proplastid stage in thepds3 mutant. Further analysis showed that high level ofphytoene was accumulated in the pds3 mutant. Addition of exogenous GA3 could partially rescue the dwarf phenotype, suggesting that the dwarf phenotype ofthepds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disrupting PDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition of many genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoene in the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cellular pathways.

Relationship between the Water Body Chlorophylla and Water Quality Factors of Wetlands Baiguishan Reservoir

[Objective] The aim was to explore the relationship between water body Chlorophyll-a and water quality factors of wetlands Baiguishan reservoir.[Method] Chlorophyll-a and water quality factors of water quality of Wetlands BaiGuishan Reservoir was studied,the analysis of the relationship on water quality of Wetlands Baiguishan Reservoir was made by use of trophic status indices and SPSS17.0 statistical analysis.[Result] Total phosphorus was an important factor of influence Chlorophyll-a in reservoir,water body had slight eutrophication phenomenon in reservoir of July to October in 2010.[Conclusion] Comprehensive management should be strengthened so as to improve the water quality of Baiguishan wetland.

Identification of Light-Harvesting Chlorophyll a/b-Binding Protein Genes of Zostera marina L.and Their Expression Under Different Environmental Conditions

Photosynthesis includes the collection of light and a/b-binding （LHC） proteins. In high plants, the LHC gene family constituting the light-harvesting complex ofphotosystems I and II. the transfer of solar energy using light-harvesting chlorophyll includes LHCA and LHCB sub-families, which encode proteins Zostera marina L. is a monocotyledonous angiosperm and inhab- its submerged marine environments rather than land environments. We characterized the Lhca and Lhcb gene families of Z. marina from the expressed sequence tags （EST） database. In total, 13 unigenes were annotated as ZmLhc, 6 in Lhca family and 7 in ZmLhcb family. ZmLHCA and ZmLHCB contained the conservative LHC motifs and amino acid residues binding chlorophyll. The average similarity among mature ZmLHCA and ZmLHCB was 48.91% and 48.66%, respectively, which indicated a high degree of diver- gence within ZmLHChc gene family. The reconstructed phylogenetic tree showed that the tree topology and phylogenetic relation- ship were similar to those reported in other high plants, suggesting that the Lhc genes were highly conservative and the classification of ZmLhc genes was consistent with the evolutionary position of Z. marina. Real-time reverse transcription （RT） PCR analysis showed that different members of ZmLhca and ZmLhcb responded to a stress in different expression patterns. Salinity, temperature, light intensity and light quality may affect the expression of most ZmLhca and ZmLhcb genes. Inorganic carbon concentration and acidity had no obvious effect on ZmLhca and ZmLhcb gene expression, except for ZmLhca6.

Deletion of a Dna K protein gene causes seedling green-revertible albino by retarding chloroplast development in rice

Green-revertible albino mutants are important sources for studying chloroplast structure, chloroplast development, chlorophyll biosynthesis, and plant photo- synthesis. In the present study, we characterized a green- revertible albino mutant gra（k）, which was obtained from the tissue-cultured rice Kitaake. The mutant gra（k） exhib- ited albino on its first three leaves. The leaf color started to turn green at the four-leaf stage. The chlorophyll contents were deeply reduced at the seedling stage, and the chloroplast development was delayed in gra（k）. The green- revertible albino （gra） phenotype of the mutant gra（k） was temperature dependent. The main agronomic traits, including plant height, tilling number per plant, seed set- ting rate, and thousand-grain weight, slightly decreased in gra（k） comparing to those in the wild-type Kitaake. Genetic analysis showed that the gra phenotype was con- trolled by a single recessive nucleic gene. By using 5,168 recessive F2 individuals derived from the cross of gra（k） × Jodan, the locus of the gene Gra（k） was delimited in a DNA region of 200 kb between the makers B-31 and P11 on chromosome 5. Sequencing analysis indicated that the three functionally annotated genes, LOC_Os05g23700, LOC_Os05g23720, and LOC_Os05g23740, were all deleted in the 200 kb region in the mutant gra（k）. Trans- genic test revealed that the gra（k） plants over-expressing LOC_Os05g23740CDS were restored to normal green as the wild-type Kitaake. Our results proved that the deletion of the DnaK protein gene LOC_Os05g23740 （encoding the chaperon protein OsHsp70CP1） led to the gra phenotype in the mutant gra（k）.