Abstract:This study was to overexpress the AtNHXS1,a new vacuolar Na+/H+ antiporter gene obtained from AtNHX1 by DNA shuffling and study its effect on the salt tolerance of rice (Oryza sativa L.).AtNHXS1 was transformed into Oryza sativa L.ssp.japonica cv.zhonghua11 by Agrobacterium-mediated method and confirmed by polymerase chain reaction (PCR) and Southern blot.The transcription level was analyzed by RT-PCR.The contents of Na+,K+ and the ability of salt tolerance were compared between transgenic lines and wild-type lines.Twenty T2 transgenic lines haboring AtNHXS1 were obtained,two of which were verified as insertion of single gene by Southern blot.RT-PCR analyses showed that the expression levels of AtNHXS1 mRNA were significantly higher after salt treatment in transgenic lines.After salt stress,it was found that the growth status,dry weight,fresh weight and Na+ content of transgenic rice plants were significantly higher than those of wild-type plants,however the accumulation of K+ was slightly low.Transgenic plants were able to survive and recover after exposure to 300 mmol/L NaCl for 5 days,whereas wild-type plants could not.These results demonstrated that overexpression of AtNHXS1 significantly improved the salt tolerance of rice.

Abstract:The full mRNA of kenaf Ms5 gene was tried to clone with RT-PCR and 3′RACE methods based on the known partial 5′-end cDNA sequence.The sequencing results showed that two transcripted cDNAs of Ms5 gene were cloned from the anther of kenaf.The length of two transcripts named as HcMs5-α and HcMs5-β were 972 bp and 1 105 bp，respectively.The structure analysis showed that both of them contained an 858 bp ORF,encoding a protein of 285 amino acids with predicted molecular weight of 32.4 ku and a isoelectric point of 7.62.The sequencing results showed that 5′-end 972 bp sequence of the two transcripts was identical,but HcMs5-β possessed an additional 133 bp sequence at 3′-end after 972 bp compared with HcMs5-α.The alternative termination of transcription in the 3′UTR of kenaf Ms5 maybe related with the post transcriptional regulation of the gene.It is the first report about the multiple transcripts of Ms5 gene in higher plant.

Abstract:Phosphomannose isomerase (PMI) gene from Saccharomyces cerevisiae was cloned by PCR,and transformed into Arabidopsis via floral dip method.The seeds were collected and germinated on selecting MS medium which mannose concentration was 1 g/L.Four positive plants were obtained.PCR for genome DNA and RT-PCR for RNA proved the PMI gene had integrated into the genome of Arabidopsis and was transcribed.The intron of yeast pmi was removed successfully in Arabidopsis.GUS activity also was detected in the transgenic plants. All the results means PMI gene from yeast can be used as a selectable marker in Arabidopsis transformation.

Abstract:Streptomyces griseoruber AL7 has been shown to display strong bioactivity against various plant pathogenic fungi and bacteria.The Escherichia coli was modified to clone host XL1-Blue MR to yield LP3,a new cloning host strain holding a Dcm DNA methylation deficient mutation and the conjugation helper plasmid pUZ8002.LP3 was used to construct the S.griseoruber AL7 genomic cosmid library with an average insert size of 40 kb.The cosmid library was then transformed into S.lividans TK24 via high-throughput conjugation to yield a heterologous expression library.A few expression clones with interesting phenotype were obtained,such as bald phenotype,precocious antibiotic production phenotype,precocious sporulation phenotype and so on.

Abstract:Endophytic nitrogen-fixing bacteria were isolated from roots,stems and leaves of rice Chujing27 collected in Lufeng County of Yunnan Province and its inhibition was identified.125 isolates were obtained by nitrogen-free medium and 15 strains were identified as nitrogen-fixers with PCR amplification of nifH gene fragments.Endophytic bacteria in different parts had different density,with peak value in roots and the lowest density in stems.Among the 125 isolates,11 endogenetic strains had suppressive effect on five main bacteria diseases, Xanthomonas oryzae pv．oryzae,Xanthomonas oryzae pv．oryzicola，Pyricularia oryzae Cav.，Thanatephorus cucumeris (Frank) Donk，Fusarium moniliforme Sheld.of rice.Three strains (NR-18,NR-64 and NR-79) isolated from roots had both functions.16S rDNA sequence analysis showed that NR-18 and NR-79 were identified as Paenibacillus polymyxa and NR-64 was identified as B．subtilis.The experiment above showed that the 3 nitrogen-fixing bacteria isolated and identified had potential value on agriculture.It was the first time that Paenibacillus polymyxa isolated as nitrogen-fixing bacteria had antipathogenic functions in the root of rice.

Abstract:Through the screening experiment and the cellulase activity measurement experiment,eight cold-adapted cellulose-decomposing bacteria have been isolated from forest humus at Heilongjiang Province for increasing the resources of cold-adapted cellulose-decomposing bacteria for composting.Among the eight bacteria,the strain B6-15 had the highest cellulase activity of 24.94 U/mL.And when the strain B6-15 was cultivated at pH 7.0,0.25% of NaCl concentration,it could get the maximum growth.Strain B6-15 was identified as a Pseudomonas sp.by its morphological,the physiological and biochemical properties and 16S rDNA sequence analysis methods.Then the thermal stability of cellulase produced by strain B6-15 was examined and the result showed that the cellulase was a kind of cold active enzyme and had 54.43% residual activity at 10℃.

Abstract:Effects of 24-epibrassinosteroids(EBR) on the seed germination and seedling growth of pepper(Huajiao 15) under the low temperature stress were investigated by both soaking method and addition method.The results showed that 0.001-1.000 mg/L EBR had no significant effect on the germinability of pepper seed,0.001-0.100 mg/L EBR significantly promoted the germination rate of pepper seed with soaking method at 0.050 mg/L with the germination rate of 44.00%.The content of EBR ranging from 0.001 mg/L to 1.000 mg/L had inhibition on the germination rate of pepper seed under addition method at 1.000 mg/L.Under both soaking method and addition method,the content of EBR ranging from 0.001 mg/L to 1.000 mg/L significantly inhibited the seedling root length of pepper seed with the highest inhibition rates at 1.000 mg/L,respectively.It also significantly promoted the seedling height of pepper seed under soaking method with 1.82 cm of seedling height,0.62 cm higher than that of control at the concentration of 0.050 mg/L,but under addition method EBR had no significant effect on the seedling height of pepper seed.Under soaking method,the content of EBR ranging from 0.001 mg/L to 1.000 mg/L had no significant effect on green weight and dry weight of pepper seedling.But there was no regularity under addition method.

Abstract:Hydroponic experiments of strawberry were conducted with three formulations of nutrient solution to select the best formulations for soilless cultivation of strawberry.Several main characteristics of growth including the fresh weight and dry weight of overground plant and root，the growth of crown，the photosynthetic and quality of strawberry，the diameter of staminate flower，the growth amass were investigated．The results showed that the effect of nutrient solution with the content of calcium nitrate tetrahydrate,potassium nitrate,ammonium dihydrogen phosphate,magnesium sulfate and ammonium nitrate at 295,303,62,246,0 mg/L on the growing status and fruit quality of strawberry are better than that of the other 2 treatments.This nutrient solution formula was the most propitious for the growth of ‘Jingyao’ strawberry and the enhancement of the fruit quality among the three different treatnments.That is,it facilitated the growth of strawberry in alimentation growth periods,and the leaves of ‘Jingyao’ strawberry was larger and thicker with a dark-green color,and the diameter of the pedicel was also larger. In addition,the nutrient solution formula was also conducive to the flowering promotion and the reproductive growth of strawberry,and resultes showed that the single fruit weight,yield and fruit quality is the best comparing to the other treatments.The results of this study provided academic and technological basis for establishing high yield and quality of strawberry in greenhouse substrate culture of irrigation and nitrogen application system. 

Abstract:Five pairs SSR primers was selected to identify 90 offsprings obtained from crossing HB pomelo(Citrus grandis Osbeck) with Jiangxi seedless Ponkan(C.reticulate Blanco).The rate of true hybrid was 97.8%.There were lost and novel bands in some of the hybrids.The genetic relationship between parents and offsprings was analyzed with UPGMA.The hybrids could be divided into four groups with similarity coefficient of 0.720 and only few progenies were clustered with parents.These created germplasms provided new materials for the citrus cultivar improvement.

Abstract:To find genes involved in the floral organogenesis of Prunus lannesiana,cDNAs for one MADS-box genes,PrseAP3 with GenBank accession No.JF710371,was cloned using homologous cloning and 3′-RACE method based on the sequence database of NCBI.The full length of PrseAP3 cDNA is 977 bp,containing an open reading frame (ORF) of 708 bp and coding for a polypeptide of 235 amino acid residues.Sequence comparison and phylogenetic analysis revealed that PrseAP3 was highly homologous to protein AP3 in Arabidopsis and belonged to the paleoAP3 clade of class B.However,semiquantitative RT-PCR analysis showed that PrseAP3 was strongly expressed in petal and stamen of single-flowered P.lannesiana cultivar ‘Makino’,and in petal,stamen and leaf-like carpel of double-flowered cultivar ‘Hisakura’,obviously different from the expression pattern of AP3 in Arabidopsis.

Abstract:Based upon the case of the hilly region in central Hunan,soil nutrient pool contents,integration indexes and Pearson coefficients among nutrient elements were studied for three typical managing target patterns,which included shoot utilizing stands (Ⅰ),shoot and timber utilizing stands (Ⅱ) and timber utilizing (Ⅲ) of Phyllostachy edulis forests.It was found that soil nutrient pool contents decreased obviously in the profiles within 0～60 cm.In different patterns and soil layers,the contents of organic matter (OM),total nitrogen (TN),available nitrogen (AN),total phosphorus (TP),available phosphorus (AP),total potassium (TK) and available potassium (AK) were 18.88-40.24 g/kg,1.08-2.19 g/kg,60.00-200.00 mg/kg,0.36-0.52 g/kg,0.84-4.07 mg/kg,11.29-23.46 g/kg and 31.72-86.70 mg/kg,respectively.In type Ⅰ,the contents of OM,TN and AK were relatively high,and the contents of AN,TP and AP were higher in type Ⅱ,the contents of TK were higher in type Ⅲ.Soil nutrient pool integration indexes was the highest in type Ⅱ,followed by type Ⅰ and Ⅲ,with score of 3.908 1,3.522 8 and 3.265 0.For the hilly region in central Hunan,the contents of soil OM,TN and AN were plentiful and phosphorus and potassium elements were in short universally in P.edulis forests.The contents of soil OM were positively correlated with TN,AN,TP,AP and AK,and there also existed Pearson coefficients between TN and AN,TP and AP.

Abstract:A nematode species,Basiria hiberna Bernard, was firstly reported in China, which was collected from the rhizospheric soil of Euonymus alatus,Sinojackia xylocarpa and Ternstroemia gymnanthera in Jiangning district of Nanjing City,Jiangsu Province.This species can be distinguished by the follow characteristics: flange-like stylet knobs; lateral field with four incisures; more posterior median bulb MB=55.3(54.6-55.8)%; 4.6(4.1-5.1) μm from dorsal gland orifice to the stylet knobs; offset spermatheca, vulva at about 61.4(60.1-64.1)% of body length; slender tail with cylindrical to clavate terminus.Euonymus alatus，Sinojackia xylocarpa and Ternstroemia gymnanthera were new hosts of Basiria hiberna Bernard.

Abstract:Using real-time quantitative PCR, the BN10 gene expression in roots, stems and leaves of rape was detected. Its expression after the treatment with methyl jasmonate (MeJA) and oxalic acid (OA) was also analyzed. The results indicated that the expression of the BN10 gene was detected in roots, stems and leaves of rape with the highest expression level in roots, whereas the lowest expression level in leaves. The expression level in the antiviral variety Suyou No.1 was higher than that in the susceptible variety Zhongshuang No.9. The BN10 gene expression was induced by MeJA and OA in a similar mode. BN10 was significantly induced after the treatment with OA and MeJA for 12 h and 24 h in Zhongshuang No.9 respectively. The expression level was 2.50-folds and 2.58-folds higher than that before the induction, followed by a decrease. After induction for 72 h, the gene expression level in the treatment of OA was 75% lower than that of the untreated control, whereas was still 1.13-folds higher in the treatment of MeJA than that of the untreated control. The BN10 gene expression in Suyou No.1 was similar to that in Zhongshuang No.9 induced by MeJA and OA excepted for that its expression level was higher than that in Zhongshuang No.9. BN10 was significantly induced after the treatment with OA for 6 h. The expression level was 3.05 folds higher than that before the induction, followed by a decrease.

Abstract:By using bioinformatics software,the domain of antigenic epitopes on Tobacco ringspot virus (TRSV) coat protein was analyzed and predicted,and the epitopes were found to be located near 319-328,384-388,420-430,480-487，495-511 amino acid residues at C-terminal. After the targeted fragment was amplified by RT-PCR based on the specific primers, the expression vector was constructed and the monoclonal antibody was prepared with the fusion protein and hybridoma. The antiserum prepared with the fusion protein reacted specifically with the expressed product of TRSV coat protein in E.coli. The antiserum had high specificity and may be applied to quarantine and inspection.

Abstract:To identify the pathogenic bacteria of kiwifruit bacterial canker, the infected kiwifruit branches and barks from Anhui (Yuexi County) Province,Shaan’xi (Huxian County) Province and Chongqing (Qianjiang County) Municipality were collected as the experiment materials. Plate streaking, gradient dilution and beef extract peptone agar (BPA) media cultivation were used for bacteria isolation, and tobacco hypersensitive responses detection, kiwifruit pathogenicity detection and 16S-23S rDNA-ITS sequential analysis were carried out as follows for bacteria identification. From the bacterial canker infected kiwifruit tissues, 10 pathogenic strains were isolated, and all isolated strains have pathogenicity and hypersensitivity to tested kiwifruit and tobacco leaves. However, different isolated strains have different pathogenicity for kiwifruit. 10 purified pathogenic strains belonged to the same pathogen through 16S-23S rDNA-ITS and Blast sequential analysis. The amplified 280 bp sequences were the same as the previously reported 16S-23S rDNA-ITS of Pseudomonas syringae pv.actinidae. It is concluded that the isolates from the canker infected kiwifruit branches and barks were Pseudomonas syringae pv.actinidae.

Abstract:The Chinese citrus fruit fly Bactrocera (Tetradacus) minax (Enderlein) is one of the most harmful pests for fruit crops. Few studies have been conducted to determine the ovipositing behavior of this species, and little is known about the movement rhythms of the adults after their emergence. In this study,the movement rhythms of B.minax adults was revealed after their emergence and the oviposition preference of females was observed through field investigations. The results indicated that the majority of B.minax adults flied around the junction zone of mixed woods and orchard in the period of over twenty days after the emergence. And it reached a peak on June 11th, we trapped (24.0±5.6) insects per pot in a 5 m zone of mixed woods in the peak period,while the date was (74.7±21.9) insects per pot in the 5 m zone of orchard, which was significantly higher than that in other trapping zones. In the peak period of emergence, more than sixty percent of the adult B.minax flied around the junction zone. After the emergence peak,the distribution of B.minax in orchard tended to be uniform. Except that no B.minax adults was trapped in the sampling zone designated as 100 m zone in mixed woods, the trapped numbers of B.minax in other sampling zones showed no significant difference. Additionally,the greatest number of oviposition marks of B.minax were found to be in the middle of an orange, which were significantly more than those in two other parts such as the stem end and the fruit navel. Furthermore, the number of oviposition marks in the dark side of orange was seven times higher than those in the sunny side. It suggested that B.minax females preferred to oviposit in the fruit night side. However,B.minax females showed no preferences to the oranges from different locations.

Abstract:The impacts of two treatments on the natural enemy community diversity was evaluated through field test. The results showed that the number of family and population of natural enemies in the plots using buprofezin and chlorantraniliprole (treatment Ⅰ) was larger than that using imidacloprid and chlorpyrifos (treatment Ⅱ). The Shannon-Wiener diversity index and evenness index of the plots with treatment Ⅰ were significant higher than those with treatment Ⅱ. The relative dominance of Lycosidae and Tetragnathidae in the plots with treatment Ⅰ was significantly higher that with treatment Ⅱ and that in the control plots. The relative dominance of Braconidae and Linyphiidae was significantly lower with treatment Ⅰ than that with treatment Ⅱ. The results suggested that using buprofezin and chlorantraniliprole to control rice planthopper and Cnaphalocrocis medinalis will be safer than using imidacloprid and chlorpyrifos. 

Abstract:The experiment was conducted to study the requirements of energy and protein of 0-4 wk Huaibei Ma chicks by investigating the effects of dietary ME (metabolizable energy) and CP (crude protein) on growing performance,body composition,and deposition of energy and protein.Eight hundred and ten Huaibei Ma chicks of 1-d-old were assigned to 9 dietary treatments following a 4-week experimental period.Nine dietary treatments were formulated to contain 10.90,11.40 and 11.90 MJ/kg ME and 19.0%,17.7%,and 16.4% CP respectively.Dietary ME,CP and interaction between ME and CP had significant effects on growing performance,body composition,and deposition of energy and protein of 0-4 wk Huaibei Ma chicks (P<0.05).Chicks fed with low-ME-high-CP diet had lower feed intake than that in other dietary treatments (P<0.01).Average daily gain was significantly higher in chicks fed with high-ME-high-CP,high-ME-normal-CP and low-ME-low-CP diets,and was significantly lower in normal-ME and low-ME-high-CP diets (P<0.01).Chicks had higher feed efficient in high-ME-high-CP treatment and had higher body fat percent in high-ME-low-CP treatment than that in other treatments (P<0.01).Highest energy deposition efficiency was found in high-ME-low-CP and low-ME-low-CP treatments,and lowest in medium-ME-medium-CP treatment (P<0.01).Highest nitrogen deposition efficiency was found in high-ME-medium-CP treatment in 0-2 weeks and low-ME-low-CP diet in 0-4 weeks,and lowest in medium-ME-medium-CP dietary treatment (P<0.01).There was no increase in nitrogen deposition efficiency as ME increased in low CP diet and as CP increased in low ME diet. In conclusion,ME and CP requirements in diet in 0-4 weeks Huaibei Ma chicks should be 11.90 MJ/kg and 17.7% or 10.90 MJ/kg and 16.4%.

Abstract:One hundred and twelve Streptococcus suis and 92 Haemophilus parasuis were isolated from pig farm to investigate susceptibility of the two bacteria strains to 23 antimicrobials by using disk diffusion method.Results showed that for the S.suis strain，the resistance rates of lincosamide，macrolides and nitrofurans were all above 90%，followed by streptomycin，gentamicin，kanamycin，amikacin and doxycycline with the resistance rates ranged from 60% to 90% and resistance rates of β-lactams and quinolones were relatively lower.For the H.parasuis strain，the resistance rate of tetracycline was the highest (78.3%)，followed by spiramycin，sulfamethoxazole/TMP，clindymycin，lincomycin，amikacin and neomycin varied from 50% to 70% and resistance rates of cephalosporins and quinolones were relatively lower.The susceptibility of the two kinds of bacteria to the antimicrobials were different，multiple resistance was severe and the resistance profiles were various.

Abstract:One year was needed for the rice-field eel,Monopterus albus,to reach their sexual maturity when cultured in cage and they were all female at the age of 1 year.The gonad index of rice-field eel cultured in cage was roughly increasing in the first year until they reached sexual maturity and the maximum of the gonad index of M.albus was 14.33±2.07,when eel reached sexual maturity,the concentration of serum estradiol reached its maximum （1 233.52±126.07 pg/mL） and the concentration of serum testosterone reached its maximum,(13.99±1.54) pg/mL,in May,a month before eel reached their sexual maturity.

Abstract:Ten microsatellite primers of Takifugu obscurus and sixteen microsatellite primers of Takifugu rubripes were used to investigate genetic diversity of four T.obscurus populations (one wild population from Changshu,one released population from Yangzhong and two cultured populations from Nantong and Shanghai respectively) in this study.Twenty microsatellite markers were amplified successfully and a total of 113 alleles were detected in the four populations.In these populations the average number of alleles was from 4.95 to 5.50,the value of average observed and expected heterozygosity ranged from 0.686 7 to 0.761 7 and from 0.653 0 to 0.700 0 respectively,and the mean polymorphic information content was from 0.601 3 to 0.638 4.Several loci were found to deviate from Hardy-Weinberg Equilibrium in each population,which indicated that the deficiency of heterozygote existed in the four populations.The value of genetic differentiation coefficient and gene flow were 0.040 6 and 5.905 6 respectively.These results suggested that the four populations have a low level of differentiation and great gene exchange among them.The genetic similarity coefficient of the four populations was from 0.859 0 to 0.915 8 and the genetic distance was from 0.088 0 to 0.151 9.Cluster analysis using the UPGMA method showed that Shanghai population formed an independent cluster and the other three populations formed another cluster.

Abstract:The ways of yolk absorption and process of organofaction in Hucho taimen,Oncorhynchus keta,Silurus soldatovi and Silurus asotus were studied using the microscope technology.The results showed that the yolks of both H.taimen and O.keta were absorbed by the blood circulation and the early digestive tract.In S.soldatovi and S.asotus,the yolk was also absorbed by the blood circulation,but the location of absorption was different.About 60-70 days were needed for the yolks of H.taimen and O.keta to be completely absorbed.The yolks of S.soldatovi and S.asotus were absorbed faster,and the yolks were completely absorbed about 3-4 days.Of the four fish species,the yolk-sphere was absorbed first,and then the fat was absorbed.The relationship between yolk absorption and organ development were discussed.

Abstract:To study the polymorphism of the proopiomelanocortin (POMC) gene and establish the molecular genetic marker for identifying different population of Pelodoiscus sinensis,populations from Taihu Lake,Huanghe River,Taiwan-introduced and Pelodiscus axenaria,were collected to get the blood.The POMC gene was amplified by RT-PCR,sequenced and analyzed thereafter.Fifteen SNPs were detected in the PCR-amplified 786 bp fragments.One SNP at nucleotide 129 in Taiwan-introduced population,one SNP at nucleotide 441 in Taihu Lake population,two SNPs at nucleotide 168 and 396 in Huanghe River population,and two SNPs at nucleotide 531 and 618 in P.axenaria were identified.The results showed that the POMC gene of P.sinensis showed polymorphic,the POMC gene of different population has specific mutation site,which can be used as a molecular genetic marker for identifying different population of P.sinensis.

Abstract:The shrimp were kept in tanks with recirculated seawater at a salinity of 3.3%.The water temperature was maintained at (29.0±1.0)℃.Hemolymph,hepatopancreas and muscle were sampled after administration of sarafloxacin hydrochloride at single dose of 30 mg/kg and the residue concentration of sarafloxacin hydrochloride analyses was performed by RP-HPLC.The results showed that,the residue concentration of sarafloxacin hydrochloride in hepatopancreas was higher than hemolymph and muscle.The residue concentration of hemolymph and muscle were very low and the former is higher than the latter.The elimination rate was markedly different in hemolymph，hepatopancreas and muscle.The elimination half-life of sarafloxacin hydrochloride was the shortest in muscle among the three tissues and the longest in hemolymph.

Abstract:The activity sounds of rice weevil Sitophilus oryzae (L.) adults, maize weevil Sitophilus zeamais (Motschnlsky) adults and rice weevil larvae in wheat,corn and coix seeds were respectively sampled. The power spectrum characteristics were analyzed and their differences in different temperatures were studied. The result shows that at 27 ℃, the frequencies of activity sound power spectrum peaks of rice weevil adults in wheat are 172.3 Hz， 409.1 Hz and 1 077.0 Hz. The frequencies of activity sound power spectrum peaks of maize weevil adults in wheat are 193.8 Hz， 430.7 Hz and 990.5 Hz. The frequencies of activity sound power spectrum peaks of maize weevil adults in corn are 236.9 Hz and 1 012.0 Hz. The frequencies of activity sound power spectrum peaks of rice weevil larvae in coix seed are 344.5 Hz and 990.5 Hz. On the other hand, the study result also shows that both the insect species and grain species affect the activity sound characteristics of grain insects, and the frequency ranges of activity sound power spectrum peaks in different temperatures are different. The experiment results provide data for the establishment of sound characteristic database of stored grain insects and the establishment of insect automatic detection.