Abstract:Dwarfism is one of the most important agronomic traits of rice.Fine mapping and functional studies of the plant height gene reveal the mechanisms for high yielding rice.In early research of this study,a main QTL Qph1 for plant height was mapped between the interval RM472-RM104 on the long arm of chromosome 1 in recombinant inbred line (RILs) population of Zhenshan 97B/Nanyangzhan.Then this QTL was used as a target fragment to construct a near isogenic line (NILs) BC3F2 whose genetic background was Zhenshan 97B.Firstly we detected the effect of Qph1 in NILs by a random population contained 320 plants,and it can explain 71.28% of phenotypic variation.Then the Qph1 was located between the interval RM6333-RM11961 through local QTL linkage analysis.In order to further reduce the target region,we planted a large genetic population which segregate in regions covering Qph1 and screened about 230 extreme recessive recombinant plants.The Qph1 locus was delimited to a 90 kb region flanked by SSR marker SHL4 and InDel marker HL13 by overlapping mapping，and compared with the sd1 has been cloned and found that they were not in the same area.It is a good foundation for cloning of the QTL.

Abstract:HJ-37 was developed through backcross and marker-aided selection by using japonica variety Nipponbare as donor parent and indica variety Zhenshan 97B as recurrent parent.F2 seeds from crossing HJ-37 with Zhenshan 97B were used to select the recombinant plants with SSR markers.Then,the homozygous seeds from the recombinant plants were used to test the plant regeneration frequency.Meanwhile,SSR was utilized to map the recombinant sites.Finally,the effect of this QTL was verified.Average of plant regeneration frequency and t value of plant 10 is 53.31% and 4.897 6.This data shows plant 10 is obviously improved in PRF.Meanwhile,the distance of QTL was narrowed to the segment between SSR markers RM3289 and RM6295.The result will be useful for isolating and cloning the gene of high frequency of plant regeneration.

Abstract:The genetic diversities of 12 weedy rices,34 cultivated rices and 36 wild rices were investigated by using 30 pairs of SSR primers,121 polymorphic bands were found,each SSR loci has 2-6 alleles,with an average number of 4.03.The genetic diversity index of weedy rice，cultivated rice and wild rice was 0.288 2、0.351 5 and 0.489 9,respectively.The average alleles of each loci were 2.1,2.27 and 3.53,respectively.This indicated that the genetic diversity of weedy rice was lower than that of cultivated rice and wild rice.The genetic distance between weedy rice and cultivated rice (0.049 4) was smaller significantly than that between weedy rice and wild rice,suggesting that the genetic relationship between weedy rice and cultivated rice was relatively closer and that weedy rice was probably degenerated atavisticly from cultivated rice.

Abstract:For further investigation of its biological functions and molecular mechanism of chloroplast heme oxygenase HO1,the recombinant protein expression vector pET28a-AtHO1 was constructed and transformed into E.coli BL21 (DE3) to express the recombinant protein.AtHO1 fusion protein was highly expressed as a soluble protein of 28-36 ku.After purification with Ni-NTA affinity chromatography and desalting,the fusion protein was used as antigen to prepare polyclonal antiserum in New Zealand rabbits.Western blot analysis showed that the antiserum could recognize the recombinant protein 6×His-AtHO1 specifically and a band between 28 ku and 36 ku of Arabidopsis and rice total proteins,indicating that the antiserum could react to the Arabidopsis and rice native protein specifically.Therefore,the polyclonal antiserum obtained by immunizing rabbits with the purified fusion protein 6×His-AtHO1 can be used for functional analysis of HO1 in rice.

Abstract:Through flask-shaking fed-batch fermentation,the optimum system of glucose feeding for poly(γ-glutamic acid) (γ-PGA) fermentation by Bacillus licheniformis WX-02 was summerized as follows：the initial concentration of glucose was 70.0 g/L,then 15.0 g/L of glucose was added at 15th hour of fermentation carried out in a 3.0 L fermentor.The results showed that the biomass and the yield of γ-PGA were 3.9 g/L and 44.5 g/L,increasing 56% and 50% compared with the control.The utilization rate of glutamic acid in the end of fermentation was 46%,while that of control was 30%.Feeding glucose at the 15th hour of fermentation process could increase biomass,enhance utilization of glutamic acid and finally increase the γ-PGA yield by B.licheniformis WX-02.

Abstract:It is important in expressing,purifying and analyzing crucial protein in vivo of solfataricus for the study of important life activities of extremely thermophilic archaea.The over-expression vector is constructed on the basis of pZC1 and pEXA vector inserted by araS promoter.Mre11 with 6×His-tag in C-terminal are expressed successfully using it.Rad50 which interplays with Mre11 is detected by co-purification.And it is ensured that the frame of the complex is still unchanged in high concentration of salt （500 mmol/L NaCl）.It indicates that the complex still keeps the function of DNA repair in adverse situation.Further analysis of co-purification makes it clear that genome DNA fragments are the essential condition of foming the MR complex.In the absence of genome DNA fragments,the molecular chaperone of archaea may bind and protect Mre11.So this expression system possesses important sense in identifying the interact network of protein in vivo.

Abstract:Rhamnose and forosamine are two essential deoxysugars components of spinosad.Four genes involved in the biosynthesis of rhamnose and forosamine,i.e.gtt (encoding NDP-glucose synthase),gdh (encoding NDP-glucose dehydratase),epi (encoding epimerase) and kre (encoding 4′-ketoreductase),are not located together with the spinosad biosynthetic gene cluster,but distributed in 3 scattered loci in the chromosome of Saccharopolyspora spinosa NRRL18395.To get an integral spinosad biosynthetic pathway,the four genes were cloned by constructing a genomic library and PCR amplifying from S.spinosa NRRL18395.The 4 genes were successively assembled into an integrative vector to construct an expression vector for biosynthesis of rhamnose and forosamine in heterologous hosts.

Abstract:The diversity of plant species of Wulipo Nature Reserve of Wushan County,Chongqing City was investigated by means of sampling,photo-taking,identifying and analyzing statistically.The results showed that there are 2 379 species (including infraspecific taxon) of spermatophyte belonging to 796 genera and 156 families in.At the generic level,temperate elements were the dominant components,in which temperate genera and tropical genera contributed 60.51% and 34.50% of the total (excluding cosmopolitan genera),respectively; disjuncted genera contributed 20.35% of the total (excluding cosmopolitan genera),in which East Asia and North America disjuncted genera contributed majorly.There are 37 genera endemic to China,contributed 4.99％ of the total (excluding cosmopolitan genera).Wulipo Nature Reserve has a special flora,which due to its unique location and entironment,that is very important to the study on middle China and south-west China flora.Compared with the flora of Sichuan and two neighboring areas,the flora of Wulipo Nature Reserve showed high similarity with Houhe Nature Reserve,exhibiting strong temperate elements.

Abstract:The kanamycin-resistant labeling strain Y2-pMarA, which has a good stability of genetics, was gained by electrotransformation of shuttle plasmid pMarA into Bacillus amyloliquefaciens subsp.plantarum B9601-Y2. Chinese cabbage was inoculated by drenching, coating seeds, soaking root and tobacco by drenching. The results show as follows: LB agar plate containing 10 μg/mL of kanamycin and PCR proved that Y2-pMarA could colonize in rhizosphere soil, root surface and inner root. After seeding for 37 d,the Chinese cabbage,under the natural soil condition,colonization numbers of rhizosphere soil, root surface,and inner root of natural soil were 94.31%, 95.00% and 84.75% of seeding for 7 d respectively; the corresponding ratios in sterilized soil were 75.26%, 84.92% and 177.74% respectively. In the Chinese cabbage with coating seeds,the ratios of colonization numbers of rhizosphere soil, root surface, and inner root of natural soil were 94.44%, 95.27% and 61.06% of seeding for 7 d respectively,while they were 75.6%, 90.9% and 69.1% in sterilized soil respectively. By transplanting Chinese cabbage seedlings soaked for 10 min, Y2-pMarA strain could expand into the soil and enter the roots of the plant. After transplanting for 33 d,the colonization density in the rhizosphere soil, root surface and inner root in the natural soil were 6.28×105, 9.54×105 and 4.69×103 cfu/g respectively; while the corresponding density were 6.97×105, 1.12×106 and 1.02×104 cfu/g in sterilized soil respectively.

Abstract:Calmodulin（CaM） plays an important regulatory role in a variety of in vivo Ca2+-dependent cell functions and enzyme systems. It was the first time to clone the calmodulin gene from Phenacoccus solenopsis Tinsley. The full-length of open reading frame (ORF) is 447 bp, encoding 148 amino acid residues. PsCaM gene was constituted by three introns and four exons. The intron lengths were 73,81，72 bp; The lengths of the 4 separated exons were 33,133,183，98 bp. Functional domain analysis showed that the protein got two EF-hand domains, with 13 Ca2+ binding sites. The theoretical protein isoelectric point is 6.21. It was stable protein, with no transmembrane region. The three-dimensional structure of its protein was obtained by homology modeling. Multiple sequence comparison revealed that the gene is relatively conservative. The research was the basis for further study of the functional mechanisms of calmodulin genes. 

Abstract:Methyl parathion hydrolyse (MPH) could degrade methyl parathion efficiently, as well as fenitrothion, ethyl parathion and chlorpyrifos, with the degradation efficiency decreasing systematically. In order to improve the degradation activity of MPH on chlorpyrifos, random mutation was carried out by error-prone PCR. Two mutants A291V and K173R were screened and expressed in E.coli BL21(DE3). The degradation activity of K173R on chlorpyrifos increased 41.55% as compared to MPH. Besides, homology modeling and molecular docking were used for preliminary analysis of the relationship between the enzymes’ structure and function. The results showed that the residue R72 played an important role in combination with chlorpyrifos, and changes occurring in the binding pocket and key amino acids eventually led to the increased degradation efficiency of K173R.

Abstract:Bacterial fruit blotch (BFB) of cucurbits, caused by Acidovorax avenae subsp.citrulli(Aac),is one of the most devastating diseases of watermelon and muskmelon. Fewer studies were done on its survival time in different places. In this study, the survival time of Aac inoculated with the anti-fampicin strain RifAac in seed,soil debit and water was investigated by isolation, bio-PCR and PCR. The results showed that Aac could survive in the watermelon seeds for 4-5 months, in the water for 2 months,in the soil for 8 months and within the soil with debris for longer than one year.

Abstract:Rice black-streaked dwarf virus (RBSDV) and southern rice black-streaked dwarf virus (SRBSDV) are the main viruses which bring rice dwarf. However, the symptoms of these two viruses are highly similar and it is difficult to distinguish them only by field observation. To construct a fast detection method of these two viruses, the PCR reaction system and program were optimized from two aspects, ie. the mixing ratios of mixed primers and the annealing temperature. Finally,an exact,sensitive and fast one-step detection method was found. At the same time, the rice dwarf virus samples from Dayu County and Nanchang County in Jiangxi Province were detected with this method, and the results revealed that all of the 10 samples from Dayu County were identified as southern rice black-streaked dwarf virus with a detection ratio of 100%, 7 out of 10 samples from Nanchang County were identified as southern rice black-streaked dwarf virus with a detection ratio of 70%, and 3 out of 10 samples from Nanchang County were identified as rice black-streaked dwarf virus with a detection ratio of 30%.

Abstract:A novel method was developed for the determination of six triazole pesticides in water samples by using ionic liquid-based dispersive liquid-liquid microextraction coupled with high performance liquid chromatography.The influence of extraction parameters including types of extraction solvent,volume of extraction solvent，extraction time,vortexing speed,NaCl concentration,pH value of sample solution were investigated.Using 100 μL 1-butyl-3-methylimidazolium hexafluorophosphate （\[BMIM\]PF6） as extraction drop,5 mL aqueous sample with 4% NaCl(pH 4) was extracted for 3 min with vortexing at 2 500 r/min.Under the optimal conditions,good linear relationships were obtained in the six triazole pesticides concentrations of 0.03-20.00 mg/L with the correlation coefficients of 0.999 6-0.999 9;the detection limits were 0.002-0.006 mg/L(S/N=3).The proposed method was applied to the determination of six triazole pesticides in tap, lake and paddy water with a recovery rate of 69.7%-101.0%,the coefficient of variation is in a range 2.7%-8.6%.

Abstract:The herbicidal activities of methanol extracts of Aralia armata (Wall.) Seem., Phytolacca americana L.,Dipsacus chinensis Batal.,Datura metel L.,Pterocarya stinoptera C. DC. and Cirsium japonicum DC. were tested against the invasive weeds,Mikaina micrantha,Bidens pilosa Lin., Conyza bonariensis (L.) Cronq, Pistia stratiotes L.and Alternanthera philoxcroides (Mart.) Griseb.. Results showed that after 7 d, at the concentration of 5.000 mg/mL, the root inhibition rates of methanol extracts of D.chinensis on M.micrantha, B.pilosa，C.bonariensis and P.stratiotes were 90.05%, 83.71%, 92.68%, 100% respectively, while the inhibition rates of A.armata on them were 79.21%, 81.17%, 88.09%,100% respectively. Methanol extracts of A.armata exhibited significant inhibition activity on P.stratiotes,C.bonariensis and B.pilosa. The IC50 values were 0.342 4, 0.409 0 and 0.696 6 mg/mL respectively. Methanol extracts of D.chinensis and P.americana showed high root inhibition effect to M.micrantha,with the IC50 values low to 0.721 0 and 0.716 7 mg/mL respectively.

Abstract:Photocatalytic degradation of sulfanilamide,sulfamethoxazole,sulfathiazole and sulfadimidine in aqueous suspension was investigated by using carbon nanotube modified TiO2 nanocomposite as the photocatalyst under visible-light and UV-light irradiation.The correlativity between molecule electronic structure and photocatalytic activity of sulfonamides was investigated by using quantum chemical method (density functional theory) DFT/B3LYP via calculating electronic structure changes.The results showed that the photocatalytic degradation rate of the four sulfonamides under visible-light irradiation was in the order of sulfanilamide>sulfathiazole>sulfamethoxazole>sulfadimidine.The photocatalytic activity of sulfonamides could be described better by their electronic structures:increasing the dipole moment and energy of HOMO and decreasing the n-octanol-water partition coefficient and the HOMO-LUMO gap lead to an increase in the photocatalytic degradation rate.The photocatalytic degradation rates of the sulfonamides in mixture relate to not only their molecule electronic structures but also the adsorption on the photocatalyst.

Abstract:Five iso-nitrogenous (45.64%±0.20% crude protein) and iso-energetic diets (16.82±0.11 MJ/kg gross energy) with graded levels of lipid at 3%(A),6%(B),9%(C),12%(D),15%(E) were formulated to feed the fish to evaluate the effect of dietary lipid levels on growth,body composition and blood biochemical indices of juvenile yellowcheek carp (Elopichthys bambusa).Three hundred fingerlings (average weight 8.26±0.81 g) were randomly distributed in five treatments in triplicate groups with 20 fish per replicate and fed with test diets twice daily for 56 d.With the increase of dietary lipid levels,weight gain rate (WGR) and specific growth rate (SGR) increased firstly,then decreased,and were significantly higher in treatment B than that in treatments A and E (P<0.05).Protein efficiency ratio (PER),with the highest level in treatment B,increased with increasing levels of dietary lipid up to 6% and showed a declining tendency thereafter.Feed conversion ratio (FCR) showed an opposite trend with PER and was remarkably lower in treatment B than that in treatments A,D and E (P<0.05).Hepatosomatic index (HSI),whole body lipid content and liver lipid content were significantly impacted (P<0.05) by dietary lipid levels,while mesenteric fat index (MFI),condition factor (CF),whole body moisture and ash contents showed no significant differences (P>0.05).HSI in treatment A was significantly lower than that in treatments C,D and E (P<0.05).The trend of whole body protein content in the experiment was similar to SGR.The levels of alanine aminotransferase (ALT),aspartate aminotransferase (AST),alkaline phosphatase (ALP) and total bile acid (TBA) in serum increased with the increase of dietary lipid levels,while total protein (TP),total cholesterol (CHO),triacylglyceride (TG),high density lipoprotein cholesterin (HDL-C) and low density lipoprotein cholesterin (LDL-C) decreased with the increase of dietary lipid contents.Analysis of SGR by quadratic model regression suggested that the dietary lipid requirement for optimal growth in juvenile Elopichthys bambusa is 7.71% (wet weight).Diet with 15% lipid could cause hepatocyte injury and abnormal fat metabolism in Elopichthys bambusa,which led to a low growth rate eventually.

Abstract:To study the relationship and the intrinsic link between the expression of liver GST(glutathione S-transferases) gene in mud carp(Cirrhinus molitorella) and Nile tilapia(Oreochromis niloticus) and the amount of cyanobacteriad,the expression of liver GST gene from the two fishes were analyzed using real-time RT-PCR and the amount of cyanobacteria intake of fish were determined in Xiangang Reservoir in different seasons.The results showed that April was the time that cyanobacteria bloomed,and simultaneously,mud carp and Nile tilapia intake the largest amount of toxic cyanobacteria.The GST genes in mud carp were expressed significantly lower in April than the rest time of the year,which suggested that mud carp may be resistant or tolerant totoxic cyanophyta.Both GSTA and GSTR2 genes in Nile tilapia had the highest expression in April.However,in the other months,the expression of GSTT was significantly high but GSTK was low in mud carp.While GSTA and GSTR2 expressed relatively low in Nile tilapia.It was obvious that GSTT of mud carp played an important role in detoxication process,but as biomarkers for the identification of toxic,it could be used only in low concentration of microcystin.The expression of GSTA and GSTR2 were positively related to the quantity of toxic algae intake in Nile tilapia.Therefore,the expression of GSTA and GSTR2 in Nile tilapia can be used as biomarkers to detect toxic algae.As Nile tilapia can consume large amounts of toxic algae,and its GST gene can detoxifytoxic algae,Nile tilapia was able to be used to biologically control of alga and finally improve water quality.

Abstract:The chromosome number and karyotype of Pseudorasbora parva were analyzed using a routine method including intraperitoneal injection of PHA and colchicine,cultivation in vivo,and slides preparation with air-drying.Whilst erythrocyte nuclear DNA content of P.parva was determined by using a flow cytometer (Beckman Coulter Inc.,USA) with serving the chicken erythrocytes (their DNA content is 2.30 pg) as reference cells.Results obtained in this study showed that:the diploid chromosome number of P.parva was 50 (2n=50); the karyotype of this species was 2n=18 m+22 sm+10 st,NF=90,while the secondary constriction,heterosome and satellite chromosome were not observed; the diploid cellular DNA contents of P.parva was 3.18 pg,which was 1.38 times to the chicken erythrocytes’.

Abstract:Viral hemorrhagic septicemia virus (VHSV),the etiologic agent of viral hemorrhagic septicemia in salmonid，caused great loss in aquaculture industry.Nucleoprotein is an important protein for activation of host immunological reaction.The nucleoprotein gene of VHSV was amplified by reverse transcription-polymerase chain reaction (RT-PCR),and cloned into pET28a vector.The recombinant E.coli BL21 (DE3) containing pET28a-N was induced and the expressed protein was detected by SDS-PAGE and Western-blot.The nucleoprotein gene fragment was 1 221 bp in length.The 48 ku target protein was expressed successfully after induced by 1 mmol/L IPTG at 5 hours.But the protein was mainly in the form of inclusion bodies after SDS-PAGE detection.The inclusion bodies was refolded with 8 mol/L urea,and diluted in 0.01 mol/L PBS (pH=9.0).Western-blot analysis showed that the expressed production can be identified specifically by the goat anti-VHSV positive serum.

Abstract:The bacterial diversity in three aquacultural systems in Shaotan river reservoir of Wuhan city was studied by the method of 16S rDNA clone library.The grass crap,silver carp,and gible carp were basal groups of the 3 aquacultural systems,while the paddlefish and big head carp were added by different proportions.No paddlefish and 6% big head carp was named system 1,while 3% paddlefish and 3% big head carp was named system 2,and 6% paddlefish and no big head carp was named system 3.The result of phylogenetic analysis showed that the sequences of 16S rDNA clone library in three aquacultural systems included 10 bacterial communities:Proteobacteria,Bacteroidetes,Cyanobacteria,Planctomycetes,Verrucomicrobia,Actinobacteria,Acidobacteria,Chloroflexi,Fusobacteria and OP10.The multifarious analysis demonstrated that the diversity index of system 2 was better than the other aquacultural systems,while the diversty index of system 3 was better than system 1.

Abstract:Persimmon tannin is abundant as a kind of natural polymer,which has strong depreciate adsorption with heavy metal ions for the rich adjacent hydroxyl groups in the B ring.The metal adsorbents made by persimmon tannin developed rapidly during recent years because of the low cost,environment friendly and high adsorption efficiency.Here we reviewed the structure characteristics of persimmon tannin,the advances of adsorbing heavy metal ions concluding radioactive elements,precious metals,toxic heavy metals and so on by persimmon tannin and the preliminary description of mechanism in adsorbing heavy metal ions by persimmon tannin.In addition,the development of the adsorption materials made by persimmon tannin is also prospected.