Abstract:Abiotic stresses are major limiting factors for crops growth and production,and stress resistance of crops can be significantly improved by overexpressing stress-related genes.In this study we aimed to isolate and identify a stress-inducible promoter which is useful for genetic engineering.Firstly a candidate promoter Oshox24P was screened and fused to GUS expression vector,and then transferred into japonica rice Zhonghua 11.GUS activity in transgenic rice plants containing GUS reporter gene under the control of Oshox24P promoter (1 918 bp in length) was induced by drought and the activity increased in response to the increasing severity of drought stress.The GUS activity was slightly induced by abscisc acid (ABA) while treatments of salt and cold had little effect on Oshox24 promoter-driven GUS expression.These results suggest that Oshox24P is a drought-inducible promoter,and will be useful for driving specific expression of target genes or proteins during drought stress,thus for genetic improvement of drought resistance.

Abstract:‘3414’ field trial was adopted to study the appropriate rate of fertilizer application and efficiency of fertilizer usage in relay cropping rice with wheat,a new cropping system in Danjiangkou region.Results showed that the yield of relay cropping rice was mainly determined by the effective panicle number per hectare and total grains per panicle.Appropriate application of N,P and K fertilizer increased yield significantly and farmer’s economic income.The highest grain yield reached up to 6 889 kg/hm2 when the rate of fertilization was N 168.0 kg/hm2,P2O5 75.0 kg/hm2 and K2O 90.0 kg/hm2.Among the effects of N,P and K fertilization,the increase of yield and economic income decreased in the order of N＞K＞P.The highest increment of yield was 3 316 kg/hm2 with an increment rate of 92.8% and the apparent nutrient efficiency of 52.9% under N application.The highest increment of yield,the increment rate and the apparent nutrient efficiency was 546 kg/hm2,8.6% and 13.0% under P application,and 699 kg/hm2,11.3% and 69.8% under K application.The modeling of fertilization effect showed that the appropriate rate of applying N,P2O5 and K2O was 188.0,30.0 and 85.0 kg/hm2 and the designable yield was 6 997 kg/hm2 in relay cropping rice with wheat in this region.

Abstract:High-temperature tolerance of two heat-tolerant rice cultivars (Ⅱ-you 838 and Shanyou 63) and two heat-sensitive rice cultivars (Ⅱ-you 63 and Teyou 559) was investigated by the HTTF (high-temperature tolerance identification of rice tiller under field conditions) method under natural high-temperature conditions between mid-July and late-August in Wuhan,Hubei Province.The results showed that the lowest seed setting rate (SSR) was observed at the hottest time point “2nd August”,and SSR of Ⅱ-you 838 and Shanyou 63 was significantly higher than that of Ⅱ-you 63 and Teyou 559,suggesting that high-temperature tolerance of cultivars Ⅱ-you 838 and Shanyou 63 was superior to that of cultivars Ⅱ-you 63 and Teyou 559,coinciding well with the practical performance of those cultivars.Similar results were obtained at time point “11th August”.The correlation and regression analysis showed that the SSR of tiller was negatively correlated with corresponding average temperature in flowering stage for all tested cultivars encountered by high-temperature stress,and the SSR of tiller of heat-sensitive cultivars Ⅱ-you 63 and Teyou 559 was much mo

Abstract:Three CIPK homologous sequences (GenBank accession number:AY104819，EU974290 and EU968441) were bioinformatically isolated from maize.Then a sequence coded by them included the motif of the CIPK family and were named as ZmCIPK1，ZmCIPK17，ZmCIPK18 based on its homologies.RT-PCR analysis showed that the expression levels of the three genes changed under the treatment of 18% PEG and 4℃.The expression of ZmCIPK1 was changed obviously in roots under the drought stress,but changed little under the cold stress.The expressions of ZmCIPK17 and ZmCIPK18 were both strongly induced.However,there are differences for the expression profiles of the two genes in different stress.At the same time,the expression profiles of the gene in leaves and roots were also different.These results showed that ZmCIPK17 and ZmCIPK18 may play a key role under drought and cold stresses with different mechanism of regulation.

Abstract:Responses of kenaf seedling to the salt stress were studied using water planting of cytoplasmic male sterility(CMS) line P3A, restorer line 992, and their F1 hybrids. The results showed that salt stress significantly inhibited the growth of kenaf seedlings. Salt stress caused changes in dry matter partitioning and increased the ratio of root to shoot. Responses to salt were different among different genotypes. The F1 hybrid showed positive midparent-/transgressive-heterosis. The heterosis of F1 increases under higher concentration of salt within certain range. 

Abstract:In this study,a 35-kb fragment was screened from the genome BAC library of a Bacillus thuringiensis strain with special primers.Sequence comparison of this fragment indicated that it contained a replicon of the plasmid p26.To test whether this fragment contain a replicon or not,a vector pHT304E that does not replicate in B.thuringiensis was constructed through deleting a 2.1-kb EcoRV fragment of pHT304 which containing the replication protein of B.thuringiensis.A 12-kb EcoRⅠ fragment cloned with the probe of replication vector pHT304E revealed that it was capable of supporting the replication in Bacillus strain.The replication region of p26 was still stable after 40 generations under the nonselective condition.

Abstract:In order to investigate the highly repetitive (HR) DNA sequences among Equus asinus genome,Cot0-Cot0.1 DNA of E.asinus genome was separated and purified through two steps of hydroxypatite chromatography.Then the HR Cot library of E.asinus was constructed by gene cloning and sequencing.After the analysis and screening of the library,18 repetitive elements were identified and submitted to GenBank,including 4 LINEs,4 SINEs,3 satellites,4 simple repeats,1 LTR retrotransposon and 2 unknowns.

Abstract:Daintain,a macrophage inflammatory factor,was characterized.To investigate the regulatory mechanism of daintain on pancreatic βcells,daintain was coupled to CNBr -Sepharose 4B,and the binding protein of daintain in pancreas was isolated by affinity chromatography,then it was characterized as cyatathionine-beta-synthase (CBS) in the database of mascot according to the peptide mass fingerprint and amino acid sequence of two peptide fragments with trpsin digestion.The interaction of daintain with CBS suggests that daintain may regulate the activity of CBS,resulting in the accumulation of homocysteine.

Abstract:The field experiments were conducted to delay the harvest cultivation of ponkan (Citrus reticulata Blanco cv.ponkan) in 2007 and 2008 using low-cost protected cultivation (plastic-film greenhouses and solar-shading nets) combined with ground plastic-film mulching.To evaluate the effect of protected cultivation on the improvement of fruit quality,the content of total soluble solid,titratable acidity,sugars and acids in ponkan fruits were measured under different cultivation models.Results showed that total soluble solid and titratable acidity of fruits from low-cost protected cultivation were higher than that of untreated control,while the volume of puffiness rate was significantly lower than that of the control.There was no significant difference in total soluble solid and titratable acidity between plastic-film greenhouses cultivation and solar-shading nets protected cultivation in the late development season of fruit.The sucrose,glucose and fructose content of all treatments increased with fruit maturity,whereas those of the control decreased in delayed harvest.Total sugars of fruits from ground plastic-film mulching in plastic greenhouses and plastic-film mulching under solar-shading nets were significantly higher than that of the control in the open field with delayed harvest,and there was no significant difference between two protective cultivations.The citric acid content during fruit maturation under different treatments shared a similar change,and the content of fruits from plastic-film mulching was higher than that of control in delayed harvest.

Abstract:The 19 CAD genes of poplar screened by bioinformatics are clustered into 3 groups.Analysis of their sequences showed that the expansion of CAD gene family in Populus was resulted from a recent whole-genome doubling and tandem replication.There are 6 paralogs in CAD gene family of Populus and Arabidopsis evolved under the purifying selection.The 19 CAD genes are expressed in all tissues of Populus.PoptrCAD17,PoptrCAD4,PoptrCAD10 and PoptrCAD7 expressed highly while the other CAD genes expressed at a lower level.Expression of the CAD genes in the same group varied with the same trends.CAD gene cloned in this research was PoptrCAD4.The catalytic activity of the CAD enzyme over-expressed in vitro were high,with the substrate preference of sinapyl alcohol.

Abstract:In the summer of 2009,fruit rot of Hylocereus undatus occurred heavily in storehouses in Zhanjiang district of Guangdong Province.The pathogen causing the disease was identified as Bipolaris cactivora based on systematic analysis including isolation,pathogenicity test and morphological observation.To our best knowledgement,it is the first report of B.cactivora infecting H.undatus in China mainland.

Abstract:Colletotrichum destructivum is the pathogen of anthracnose in Anthurium andraeanum. Based on internal transcribed space (ITS) sequences of Colletotrichum genus, a pair of specific primers (F1 and ITS4) to detect C.destructivum was synthesized. The primer sets amplified a single PCR band of 486 bp with DNA extracted from C.destructivum isolated from A.andraeanum,while other relative strains within different species had no corresponding band. The detection sensitivity was 10 pg of genomic DNA. When using ITS1/ITS4 as the first round primes and F1/ITS4 as the second round primes, the detection sensitivity increased 10 000-fold to 10 fg. The detection sensitivity for the soil pathogens was 200-conidia per gram soil. The PCR-based method developed here could stably and quickly detect the pathogen from water samples and diseased plants.

Abstract:Banana fusarium wilt,caused by Fusarium oxysporum f. sp.cubense race 4 (Foc4), is one of the most destructive diseases of banana worldwide. To obtain the toxin of Foc4 effectively and provide the foundation for further research, the culture filtrates of Foc 4 obtained from different culture media,in different incubation modes, with different incubation time, incubation temperatures, initial pH values of Richard medium, rotation speeds of shaker, and inoculation amounts, under different illumination conditions were tested for their bioactivities to banana plantlets in vivo by using root-dipping method. An optimized system for toxin production of Foc 4 was established, in which the optimal condition for Foc 4 to produce toxin was obtained when it was cultured in Richard medium with an initial pH 7-9 and 5 mycelial plugs per 300 mL,in an incubation temperatures of 25-30 ℃ under continuous illumination for 24 h in a shaker at 140 r/min for 9-12 days.

Abstract:The experiments were conducted to determine the debilitation effect of Spodoptera litura nucleopolyhedrovirus (SlNPV) on hosts from the infected parental generation to their next two progenies through feeding the larvae with the concentration of 9.44×107 OB/mL and the adults with the concentrations of 9.44×108 OB/mL and 9.44×107 OB/mL in the laboratory. The experiment results of feeding the early 4th-6th instar larvae with SlNPV in concentration of 9.44×107 OB/mL indicated that the pupation rates of F1 and F2 progeny were respectively reduced by 12.73%-18.59%, 5.88%-10.21%, and the emergence rates went down respectively done by 11.31%-15.41%, 5.70%-8.46%. The experiment results of feeding the adults with SlNPV in concentrations of 9.44×108 OB/mL and 9.44×107 OB/mL showed that the pupation rates of F1 and F2 progeny were respectively reduced by 14.00%-34.63%, 8.33%-9.84%, and emergence rates were respectively decreased by 10.77%-19.61%, 11.36%-12.00%. After feeding the early 4th-5th instar larvae and adults with SlNPV,pupal weight and fecundity of F1 and F2 progeny were both significantly decreased, oviposition period and adult longevity were both significantly shortened,but there were was no significantly difference in larval duration, pupal stage and preoviposition period between F1 and F2 progeny. The results also indicated that the debilitation effect of SlNPV on host was weakened gradually with the increase of host progeny.

Abstract:The study aimed to build the plant volatile organic compound database (PVOCD), investigate the relationship between PVOCs and insect behavior and provide theoretic basis for the development of new pesticides. The PVOCD was constructed on the basis of MDL ISIS Base scientific information management system and its information was collected from papers published at home and abroad. A virtual screening was carried out on the PVOCD by Discovery Studio. The PVOCD contained 2 410 compounds, including molecular formula, structure, molecular weight, CAS registry number and so on. 6 candidate compounds for general odorant binding protein Ⅱ of Spodoptera litura (SlitGOBPⅡ) were screened out from PVOCD by molecular docking, and prednisone was the best one binding with SlitGOBP II. The construction of PVOC and the application of molecular docking could provide theoretic basis for the development of new pesticide.

Abstract:The field demonstration experiments of rice leaf roller and striped rice borer with Chlorantraniliprole 20% SC were conducted in Jiangxi,Zhejiang,Anhui,Hubei,Hunan,Jiangsu,Sichuan Province from the year of 2008 to 2009 in China. The results indicated that the range of insecticidal activity and leaves protecting effects towards rice leaf roller were 90.2％-100.0％ (for 7 days after treatment) and 82.9％-96.8％(for 14 days after treatment) respectively,which were higher than those of fipronil 5% SC. The range of insecticidal activity and seedling (or spike) protecting effects towards striped rice borer were 89.0％-96.9％ and 82.8％-96.0％ (for 21 days after treatment) respectively,which were higher than those of fipronil 5% SC and chlorpyrifos 48% EC. The results of field demonstration experiments revealed that Chlorantraniliprole was more effective in the control of rice leaf roller and striped rice borer,which can be promoted in practice and meanwhile,the application technologies in the large field areas were provided. 

Abstract:Pot experiment was conducted to study the effects of spraying different micronutrients and amino acids into the surface of leaves on yield and quality of Chinese cabbage (Brassica rapa L.).The results showed that methionine,zinc,molybdenum,boron and glycine could increase yield of Chinese cabbage.The treatments of spraying methionine,zinc and molybdenum increased yield of Chinese cabbage significantly.All treatments could reduce the nitrate content of Chinese cabbage with the highest decrease of 35.0% compared with the control.Molybdenum,boron,zinc and methionine might reduce nitrate content of Chinese cabbage significantly.Experiment also indicated that glycine,proline and selenium could both increase the contents of vitamin C,soluble sugar and soluble protein of Chinese cabbage.Therefore,spraying micronutrient or amino acids into the surface of leaves is a good application for increasing yield,reducing nitrate content and improving quality of Chinese cabbage.

Abstract:Diphenylpicrylhydrazyl (DPPH) spectrocolorimetery method was used to evaluate the anti-oxidative capacity of phytic acid,ascorbic acid (ASA),glutathione (GSH) and vegetable extracts.The results showed that eradication of free radical of phytic acid,ASA and GSH was pH dependent.The IC50 ,a value describing the ability of DPPH free radical scavenging,was 1.94 ×10-2(pH=2.0),0.26 (pH=4.6) and 1.86 (pH=5.7) for phytic acid,GSH and ASA,respectively.In addition,adding phytic acid enhanced the eradicating ability of free radical of vegetable extracts.The results suggested that phytic acid would be an important antioxidant in plants due to its high content in plants,and pose a crucial role in resistance to oxidative stress.

Abstract:SRAP (Sequence-related amplified polymorphism) markers and mitochondrial DNA control region sequencing were used to investigate genetic structure of four loach (Misgurnus anguillicaudatus) populations from two sympatric distribution areas of natural diploid and tetraploid loach-Dongting Lake (DT) and Wuhan city (WH) in China.The results from SRAP showed that a total of 534 loci were detected in four loach populations by 18 pairs of polymorphic primer combinations,and the numbers of amplified loci per primer pair ranged from 23 to 40.The averages of Nei’s gene diversity (h) and Shannon’s information index (I) of each population were from 0.205 to 0.218 and from 0.324 to 0.341,respectively,and there were no significant difference among h-values or I-values of four populations.UPGMA dendrogram tree,which was constructed based on the Nei’s unbiased genetic distances,showed that DT diploid and DT tetraploid populations were firstly clustered into one branch and their relationships were the nearest,then WH tetraploid population was clustered to the branch.And the other branch contained only WH diploid population.Partial sequences of mitochondrial control region (932-935 bp) of 40 individuals representing four loach populations were sequenced.A total of 47 nucleotides were variable,resulting in a total of 29 haplotypes.Nucleotide diversities (π) of DT tetraploid and DT diploid within populations were 0.898% and 0.872%,respectively,and they were significantly larger than those of WH tetraploid (π=0.465%) and WH diploid (π=0.675%).Within the sympatric diploid and tetraploid loach populations,genetic diversity of DT tetraploid was larger than that of DT diploid,and the genetic diversity of WH diploid was significantly larger than that of WH tetraploid.Analysis of molecular variance (AMOVA) indicated that the genetic variation mainly occurred within populations (63.55%),and 37.32% of variation occurred among populations.Significant population subdivision was supported by both pairwise FST values and Kimura 2-parameter genetic distances between populations,except for no apparent subdivision between DT tetraploid and DT diploid.Phylogenetic tree through Bayesian inference from the control region sequence haplotypes was similar to the genetic relationship among four loach populations revealed by SRAP analysis.

Abstract:Channel catfish virus was propagated in the channel catfish ovary cell line and then purified by ultracentrifugation.This purified virus was used to immunize the female BALB/C mice to gain polyclonal antibody.ELISA was used to detect the antibody titer.The bioactivity was identified by indirect immunofluorescent assay and serum neutralization tests.The ELISA titer of the polyclonal antibody was 1∶25 600.The polyclonal antibody showed high specificity at the indirect immunofluorescent assay.It could stop the virus from infecting the cell efficiently.

Abstract:Heat shock protein 60 (HSP60) functions as a molecular chaperon and plays an important role in protein folding,maintenance of structural integrity and proper regulation of a subset of cytosolic proteins.To identify a sensitive biomarker of freshwater monitoring,the full-length cDNA of Tanichthys albonubes HSP60 (designated TaHSP60) was cloned by RT-PCR and RACE techniques.It was of 2 486 bp,including 5′UTR of 102 bp and 3′UTR of 656 bp.Its open reading frame contained 1 728 nucleotides which encoded a 575 amino acid peptide.The deduced amino acid sequence of T.albonubes HSP60 had the highest similarity with Danio rerio (96.2%).The similarity between T.albonubes and Carassius auratus,Paralichthys olivaceus,Salmo salari and Xenopus tropicalis was 93.2%,89.7%,88.3% and 83.8%,respectively.Clustal X analysis confirmed the existence of the typical mitochondrial signature sequence,ATP binding region and conserved GGM repeat motif at the C-terminal in T.albonubes HSP60.Phylogenetic analysis placed T.albonubes and the putative D.rerio HSP60 into one separate cluster.The results from real-time PCR showed that the T.albonubes HSP60 was ubiquitously expressed in different tissues such as liver,muscle,gill,fin clips,eye,ovary,intestine and brain. HSP60 expression levels in liver were the highest while extremely low in gill and fin clips.Statistical analysis indicated that the transcription of HSP60 in liver was significantly higher (P<0.05) than in any other organs.After copper exposed,mRNA expression level of TaHSP60 in liver were significantly higher than those in control group in 48 h and 96 h(P<0.05).The data would help design nucleotide probes for detecting HSP60 gene expressions as a biomarker in environmental monitoring.

Abstract:Histopathology and ultrastructural pathology of Scylla Serrata infected by reovirus were observed by paraffin section and ultrathin section technique.The results of histopathology examination showed that the main histopathological changes found in hepatopancreas,gills,intestin canal,heart,stomach and muscle tissues.Among them,hepatopancreas,gill and intestine were the main target tissues with the most obvious pathological changes,which included damaged basal membrane of hepatopancreas,hepatocyte necrosis,clogging gill cavity,shedding epithelium of intestine，muscular fibers derangement,and fragmentation.The results of ultrastructural pathology examination showed that SsRV widely distributed in liver,pancreas,gills,heart tissue and could reproduce in these organs.Therefore,SsRV virus apparently could infect a variety of target organs.Moreover,SsRV infection caused a series of pathological changes in the target tissue cells,such as the condensation of nuclear chromatin,the increasing of electron density,the formation of myelin bodies and lysosome,the destruction of mitochondrial structure and the expansion and distortion of rough endoplasmic reticulum.

Abstract:The existing research papers have demonstrated the strong immunogenicity of outer membrane proteins to aquacultured creatures.Using the previously constructed recombinant plasmid pET30a-OmpK as template,the peptide coding sequence of OmpK was amplified by PCR.After introducing restriction sites at both 5′ ends and 3′ends of the sequence,the gene was cloned into secreted expression vector pPIC9K,and formed the recombinant vector pPIC9K-OmpK.The vector was linearized bySalⅠ,and transformed into Pichia pastoris strain GS115 by the method of Lithium Chloride transformation.Transformants were selected by G418 and confirmed by PCR.The recombinant protein was expressed and secreted into the supernatant after inducing by methanol.SDS-PAGE analysis indicated the recombinant protein OmpK was hyperglycosylated with a molecular weight about 37 ku,larger than the expected size 29 ku.Western-blotting results showed that the recombinant protein OmpK could react with the antiserum against OmpK expressed by E.coli BL21.It indicated that the protein OmpK was glycosylated but remained its antigenicity.

Abstract:To improve the quality and meet the requirements of sowing and planting, the production testing and demonstration have been done for stubble-mulch rotocultivator for boat tractor in different paddy fields with multiple rice cropping system. By upgrading core technology and integrating matching technology in the production testing and demonstration process, the technical and economic indexes of stubble-mulch rotocultivator for boat tractor have met expectations, a new conservation tillage technology for rice production mechanization has been formed and it has been used to achieve moderate tillage and stubble-mulch returning. A kind of mechanization technology system for cultivating, seeding and harvesting of paddy has been created, and it uses the stubble-mulch rotocultivator for boat tractor as the prime technological device.