Abstract:Effects of deeply mechanized fertilizer application on the quality and grain yield of direct seedling rice were studied using super hybrid rice Peizataifeng and super inbred rice Yuxiangyouzhan as samples in field trials.The results showed that grain yields were increased significantly when the increase of rice special fertilizer rate ranged from 0 to 1 500 kg/hm2.The number of effective panicle per m2 and solid grain per panicle were increased.More application of rice special fertilizer would increase plant height,index of leaf area,SPAD in sword leaf at late stage,and total dry mass weight of rice.The results indicated that deeply mechanized fertilizer application can improve quality of rice significantly.

Abstract:The nodulation receptor kinase (NORK) of Astragalus sinicus mainly contains three functional domains including repetitive sequences of the N-terminal leucine-rich domain (LRR),the central transmembrane domain (TM),and the C-terminal kinase domain (PK),respectively.Using yeast two-hybrid technique and TM domain as the bait protein,its interacting target proteins were screened and identified.41 original positive clones were obtained.After the inserted cDNA sequencing and Blast analyses in the NCBI database,an interacting target protein with the same biological function as a translation elongation factor protein EF1α was identified.The corresponding protein encoding gene was named AsEF1-α.The further confirmed experiment showed that the key interacted domain of AsEF1-α protein was located in the EF1-α-Ⅲ.These results provide a fundamentally theoretical basis for future in-depth study of the genes’ function and mechanism,and also provide novel information and material for establishing early root nodulation signaling network in legume plant.

Abstract:A putative polyketide synthase (PKS) gene sahA was found in the Streptomyces sahachiroi genome by PCR using degenerated primers for the ketosynthase alpha(KSα) gene.The phylogenetic tree and blast analysis revealed that sahA was belonged to the type Ⅱ PKS gene family and closely related with the subgroup of spore-pigment biosynthetic PKS genes.Once sahA was disrupted by single crossover,the spores color of Streptomyces turned into white from leaden but the time of spore-forming,quantity of sporulation,UV-resistant ability of spores and azinomycin B production had no change.It demonstrated that the putative type Ⅱ PKS gene sahA might be involved in the biosynthesis of S.sahachiroi spore-pigment.

Abstract:To provide theoretical bases for improving yield and quality of cultivation,regulating flower time and breeding efficiently,the morphological observation,paraffin section and semithin section were used to investigate the process and characteristics of the flower bud differentiation and anther development of French marigold (Tagetes patula L.) at both spring and autumn.The results showed that French mariglod had a typical capitulum consisted of ray florets and disk florets.The differentiation of flower bud was not beginning until the first two pairs of leaf primordium differentiation.The differentiation process of inflorescence was consistent with the sequence as follows:inflorescence primordium differentiation phase,bract primordium differentiation phase,ray floret primordium differentiation phase,disk floret primordium differentiation phase,ray floret differentistion phase and disk floret differentistion phase.There were differences in the initial and duration time of the differentiation of flower bud in different species and at different seasons,showing earlier time at autumn.The process of anther development was as follows:archesporial cell,sporogenous cell,microspore mother cell,dyad,tetrad,microspore,and matured pollen grain.The tapetum is amoeboid tapetum and the matured pollen grain is tri-cellular pollen.

Abstract:Based on plot data of shoot utilizing stands (Ⅰ),shoot and timber utilizing stands (Ⅱ) and timber utilizing (Ⅲ) of Phyllostachy edulis forests located at the hilly region of central Hunan,quantities of soil microbe,enzymatic activities and their canonical correlation analysis (CCA) were investigated in the three typical patterns of management.Results showed that bacterium was the major kind of the total soil microbe with the highest proportion of 84.51%-91.31%,followed by fungus and actinomyces with the proportion of 8.39%-14.84% and 0.17%-0.65%,respectively.Within 0-60 cm soil layers the highest quantities of bacteria,fungi and actinomycetes occurred in type Ⅰ that were 83.68×106 cfu/g,8.27×106 cfu/g and 0.26×106 cfu/g,respectively.The variable activities of soil urease,proteinase,sucrase,catalase and polyphenol oxidase were 0.008-0.027 mg/g,0.019-0.026 mg/g,1.23-2.46 mg/g,0.28-0.46 mL/g and 0.35-1.57 mg/g in soil layer of 0-60 cm,respectively.The highest activities of urease appeared in type Ⅲ.Activities of proteinase,sucrase and catalase were the highest in type Ⅱand the highest activities of polyphenol oxidase were in type Ⅰ.Through canonical correlation analysis(CCA),the canonical variable (U) was positively related with fungus number (1.264 5) and negatively related with bacteria number (-1.091 8).The canonical variable (V)was negatively related with sucrase activities (-0.876 7).

Abstract:In this paper, we reported results about characterization of the factors affecting Agrobacterium tumefaciens-mediated transformation (ATMT) of the plant pathogenic fungus Botrytis cinerea strain RoseBc-3 and about the validity of ATMT for the particular strain of B.cinerea. The results showed that the temperatures for co-incubation of A.tumefaciens and B.cinerea and the conidial concentration of B.cinerea in the co-incubation mixture greatly affected the transformation efficiency. The optimal co-incubation temperature was 22 ℃ and the optimal conidial concentration was 1×105conidia/mL. Under the optimal co-incubation conditions, the numbers of transformant reached 100 per dish (9 cm diameter). All the 16 randomly-selected transformants of B.cinerea were confirmed to harbor the T-DNA of A.tumefaciens based on the result of the specific PCR. The result of the Southern blotting further indicated the integration of the T-DNA in the genome of B.cinerea with the frequency of the single-copy insertion reaching 87.5%. Six types of mutants were identified from the transformants. They were sporulation deficiency mutants (3.8%), mycelial growth-reduced mutants (3.5%), sparse mycelia mutants (1.6%), colony color mutants (5.7%), pathogenicity decreasing mutants (3.3%) and pathogenicity enhancing mutants (3.0%). DNA sequences flanking the T-DNA tag were obtained using the inverse PCR technique and the hi-TAIL PCR technique from 14 mutants of B.cinerea. The insertion sites in these mutants were located with the same frequency in the coding region and in the non-coding regions.

Abstract:Cg-1,with 8 homologous genes,from Meloidogyne javanica is a candidate avirulence gene corresponding to tomato resistance gene Mi-1. Comparing with its homologs,Cg-1 contains a CAATGA fragment insertion. In this study, Cg-1 homologous clones from Meloidogyne enterolobii,a root-knot nematode that is able to overcome tomato Mi-1 resistance, were firstly cloned. Meanwhile,Cg-1 and its homologous clones of an M.javanica avirulent strain in our laboratory were cloned. Comparison of Cg-1 and these homologous clones from M.enterolobii and M.javanica strains showed that Cg-1 gene was inexistent in M.enterolobii that is there were only Cg-1 homologous genes which did not contain CAATGA existing in M.enterolobii. This result further indicated that Cg-1 may be an avirulence gene. Sequence analysis of Cg-1 gene predicts that the third open reading frame (ORF) encoding peptide and its structural mRNA,or both exercise biological function of Cg-1.

Abstract:Weed survey was conducted by inverted W-pattern to determine the species composition and structure of weed communities in rice fields in Hubei Province in China. The results showed that 138 weed species (including varieties) belonging to 46 families and 94 genera were found. Among them,6 species were considered as worst weeds, including Echinochloa crusgall,Monochoria vaginalis,Leptochloac hinensis,Eclipta prostrata,Alternanthera philoxeroides and Cyperus difformis; 11 species were regional worst weeds; 24 were common weed species and 97 were normal weed species. The worst weeds were the main components of weed communities in all rice regions in Hubei, the typical weed communities were E.crusgalli+E.prostrata+A.philoxeroides,E.crusgalli+L.chinensis+E.prostrata,and E.crusgalli+M.vaginalis+C.difformis. The richness and diversity species of weed community in paddy rice fields in North Hubei region were higher than other regions, but the evenness index of Central-east Hubei region were relatively lower. Based on the Sorensen′s similarity index, the structure of weed community in North Hubei region was more similar to that of the overall communities in Hubei. The difference of weed occurrence and community structure might result from the cropping system, tillage type and environmental conditions.

Abstract:The inhibition activities of tea saponin against Pyricularia grisea,Rhizoctonia solani,Sclerotinia sclerotiorum,Penicillium italicum,Glomerella lagenaria,Fusarium oxysporumr,Glomerella ciugulata,Colletotrichum musae,Phytophthora parasitica,Fusarium oxysporum,Bipolaris maydis and Peronophythora litchi were studied by mycelium growth rate inhibition assay. The results showed that tea saponin had significant antifungal activity. After treatment with 1 000 μg/mL, the inhibition rates of tea saponin against Pyricularia grisea,Penicillium italicum,Glomerella lagenaria,Sclerotinia sclerotiorum and Bipolaris maydis were 100.00%, 100.00%, 89.12%, 87.27% and 83.13% respectively. The EC50 values against Pyricularia grisea and Penicillium italicum were 20.15 μg/mL and 11.39 μg/mL respectively. Tea saponin had no activity against Fusarium oxysporum with the EC50 value exceeding 2 000 μg/mL. On the other hand,tea saponin had higher antifungal activities against Pyricularia grisea than sulfur tricyclazole,and the inhibition rate against Sclerotinia sclerotiorum was not significantly different betweent tea saponin and procymidone by toxicity comparison in laboratory.

Abstract:Soil structure is the material base of soil fertility and has an important significance in soil fertility.In order to study the relationships between soil particles and the fertility of black soil in northeast China,fractal dimensions of soil inorganic particles and complex were calculated by the particles fractal theory.The distribution characteristics of black soils in northeast China and its relationships with soil fertility were analyzed as well.The results showed that the fractal dimensions of black soil inorganic particles in northeast China were 2.80-2.90,significantly correlated with the entire size fraction.The fractal dimensions of complex were 2.66-2.93,correlated positively with clay and negatively with silt complexes.The regression equation（y=88.18x－60.73x2+10.57x3）between the fractal dimension of inorganic particles and complex showed that inorganic particles and complex organic could decrease the fractal dimension,and might decrease soil complex degree because of low temperature and increase the fractal dimension of soil.Association analysis between the fractal dimension of soil particles and the fertility confirmed that inorganic particles and complex organic could increase soil nutrients storage capability,and might release nutrients gradually in the process of mineralization.It can be concluded that the fractal theory could be taken as a quantitative indicator explaining the mechanisms of inorganic compound particles and soil structure and indicating fertility characteristics under the conditions of the fractal dimension of soil.

Abstract:Ephemeral gully plays an important role in soil losses in hilly red soil region.A field experiment was carried out to study the impact of contour hedgerow on spatial variation of soil water in red soil slope-land with ephemeral gullies.The results showed that there was an upward trend for the soil moisture from the soil surface to the bottom in the dry season.However,there was an upward trend for the soil moisture from the soil surface to the bottom in plot in the rainy season without contour hedgerow setting while it shows that the trend was 30 cm soil layer>45 cm soil layer>15 cm soil layer in the plot with contour hedgerow setting.The coefficient of variation of soil moisture in plot with contour hedgerow setting showed a decreasing tendency from the soil surface to the bottom,and it is larger than that of plot without contour hedgerow setting in the same layer.In the wet and dry seasons,the soil moisture in plot with contour hedgerow setting is more than that of plot without contour hedgerow setting at slope middle,but less at upper slop and lower slop.Therefore,the contour hedgerow can maintain soil and water effectively,making the plants to take advantage of shallow slope water in the wet and dry seasons,which plays a significant role in easing seasonal drought and conserving soil and water.

Abstract:The differences of Mo absorption and its relationship with root morphology between Mo-efficient winter wheat cultivar 97003 and Mo-inefficient winter wheat cultivar 97014 were studied with nutrient solution culture.The results showed that the accumulation of Mo at shoots of cultivar 97003 was higher than that of cultivar 97014 under Mo deficiency.In addition cultivar 97003 had higher ability of transferring Mo to leaves than that of cultivar 97014.Under two Mo levels,Mo accumulation in leaves and the whole plant of cultivar 97003 was 86.2% and 96.3%,higher than that of cultivar 97014,which was 78.9% and 87.2%.The root morphology of two winter wheat cultivars under Mo deficiency were different,cultivar 97003 had good root morphology.The root length,surface area,volume and average root diameter of cultivar 97003 were 1.16,1.41,1.21,1.73 times higher than those of 97014,surface area,volume and average root diameter were all significantly higher than those of 97014.The root length,surface area,volume and average root diameter of cultivar 97003 were 1.13,1.28,1.12,1.44 times higher under Mo deficiency than those in providing Mo treatment.Expected root length,surface area,volume and average root diameter were all significantly higher than those in providing Mo treatment.In contrast,root length,surface area,volume and average root diameter of cultivar 97014 had no significant difference whether providing Mo or not.

Abstract:Using cropping soil (Shandong Shouguang pepper greenhouse) as basic matrix and the chemical fertilizer as control,five different organic fertilizer formulations with the same level of N,P and K content were applied to pepper plants potted in soil from a continuous cropping greenhouse to compare impacts of organic fertilizers on rhizospheric microbial diversity and the growth of peppers.The rhizosphere microbial quantity and diversity were investigated using random amplification of polymorphic DNA (RAPD) and a traditional plate counting method.Results showed that most types of organic fertilizer can increase rhizosphere soil microbial diversity and the organic fertilizer made from silk worm excrement plus yeast fermentation waste (SYM) is the highest one.SYM provides the best results in promoting plant growth,protection of seedlings from damping-off,and improving the vitamin C and capsaicin content of fruits.These results indicated that organic fertilizer produced by fermented waste is conducive to the sustainable development of agriculture which can increase yields and quality of crop,improve micro-ecological structure of soil in continuous cropping.

Abstract:Taking rape fertility information of Hubei Province as an example,the Geoprocessing model was established using fertility analysis process of Kriging method.With update modeling of the soil nutrients data,the method of reconstructing the Geoprocessing model will be further elucidated.Geoprocessing in rape fertility can improve efficiency of analysis.

Abstract:The ten-day change of meteorological factors from different regions of Hubei Province since 1981 and its effects on growing stage change of mid-season rice were analyzed,which was based on meteorological data including growing stage of mid-season rice,average temperature,the average maximum temperature,the average minimum temperature,the sunshine duration and the rainfall from 1981 to 2008 taken from national basic meteorological observation stations of Hubei Province including five regions as Wuhan,Suizhou,Lichuan,Zhongxiang and Gucheng with complete observation data of mid-season rice.The results showed that the average temperature,the average maximum temperature and the average minimum temperature exhibited an increasing tendency in the vegetative growth of mid-season rice.The average maximum temperature exhibited an increasing tendency in the reproductive growth,but the average temperature and the average minimum temperature showed a decreasing tendency.Moreover,there was an obvious decreasing tendency for the average maximum temperature in the midmonth of August.The sunshine duration showed a decreasing tendency in the whole growth stage.The growing stage would shorten with the average temperature and the average minimum temperature increasing in the full-growing stage and vegetative growth.However,its decreasing would result in extending the growing stage of mid-season rice.The days of full-growing stage and vegetative growth of mid-season rice exhibited an extending tendency in the southeast,southwest,northeast and northwest of Hubei Province influenced by the change of comprehensive meteorological factors,but a shortening tendency in Jianghan Plain.The days of reproductive growth showed an extending tendency in the five regions mentioned above.

Abstract:To better understand the biological characteristics of Luxi cattle embryo skeletal muscle satellite cells,those cells from the skeletal muscles of the limbs of 30-50 days old Luxi cattle embryo were isolated and cultured.The results showed that two surface antigen markers,Desmin and Myod,were positively expressed in the harvested cells by immunofluorescence.Desmin,Myf5,c-Met and pax7 were found to be positively expressed by RT-PCR.Growth curve indicated that the cells were capable of proliferating strongly.In addition,the cell cycle analysis showed that the cells in S phase accounted for 8.94% of the total.After 10 days for myoblasts induce,the maker specific for fast skeletal myosin of myotubules was positively detected by immunofluorescence.After 7 days for adipogenic induce,lipid droplets were detected to be positive using Red Oil O dye; the adipocyte specific genes,ppar-γ and LPL,were identified to be positively expressed by RT-PCR.The Luxi cattle embryo skeletal muscle satellite cells were cultured successfully via vitro culture system,and could be induced to differentiate into multinucleated myotubes and adipocyte,respectively.This provided a new method for preserving animal genetic resources and for study stem cells theoretically in the future.

Abstract:Grass carp reovirus (GCRV) has a genome consisting of 11 double-stranded RNA,and possesses several structural proteins.The VP5 protein is one of the structural proteins in the core capsid encoded by the M5 gene of GCRV-GD108,a reovirus strain which was isolated from grass carp cultured in Guangdong and had been identified by our lab recently.Primers with restriction enzyme digesting sites were designed based on the cDNA of M5 gene.The amplification products of M5 gene were digested and cloned into prokaryotic expression vectors.Two recombinant protein expression vectors,pET30c-M5 and pColdⅡ-M5,which had been identified by PCR,enzyme digestion and sequence analysis,were transformed into E.coli strains,respectively.Then the recombinant expression strains were induced.The effects of culture medium,induction time,temperature and IPTG levels were analyzed,and suitable conditions for the expression of the target protein were proposed.The results of SDS-PAGE and Western blot showed that the target protein VP5 with molecular weight around 80 ku was obtained,but the recombinant protein was mainly presented in inclusion bodies.The inclusion bodies were then denatured,dialyzed and renatured to get the recombinant protein.The target protein,which was expressed by pET30c-M5/BL21(DE3) accounted for 22.5% of the total bacteria proteins,and was highly purified (>95.0%).The pET30c-M5 is considered to be an engineering strain suitable for large scale expression for it cost less time of inducing and got higher production than pColdⅡ-M5.

Abstract:Soluble glutathione S-transferase (sGST) is extremely important for metabolic detoxication by catalyzing the conjugation of GSH (glutathione) with microcystins.In this study,the amino acid sequence of a mu-class sGST cDNA fragment from the liver of Macrobrachium nipponensis were firstly obtained by RT-PCR using degenerated primers.Sequence analysis revealed that the cloned mu-class sGST cDNA fragment was 299 bp in length,encoding 99 amino acids.Homologous analysis showed that the nucleotide sequence of the glutathione S-transferase gene in M.nipponensis was 60% homologus to the glutathione S-transferase gene of Litopenaeus vannamei,Rattus norvegicus,Mus musculus,Ixodes scapularis,Rhipicephalus appendiculatus,Xenopus tropicalis,Rhipicephalus microplus and Crassostrea gigas.This suggested that the sGST cloned in this study belonged to mu-class GST.The result of further experiment showed that mu-class sGST level did not change significantly in M.nipponensis exposed in MC-LR.

Abstract:A growth trial was conducted to evaluate the optimum requirement of dietary phosphorus for juvenile red swamp crayfish,Procambarus clarkii.Triplicate groups of juvenile red swamp crayfish (initial mean weight:5.02±0.51 g,mean±SD) were fed diets containing different levels of phosphorus (0.83%,1.07%,1.25％,1.38％,1.59％,1.82％ and 2.03％) for 70 d.The results showed that both the weight gain and specific growth rate increased significantly with increasing dietary phosphorus up to 1.82% and then decreased beyond this level.Feed conversion ratio decreased significantly with increasing dietary phosphorus level up to 1.82%.Dietary treatments did not significantly affect survival rate.Body composition analysis showed that the whole-body ash and phosphorus contents were significantly affected by dietary phosphorus levels.Dietary phosphorus levels also had distinct effects on hepatopancreas alkaline phosphatase activity.Broken-line analysis based on weight gain and specific growth rate indicated that the minimum phosphorus requirement for the optimal growth of juvenile red swamp crayfish were 1.84% and 1.80%,respectively.

Abstract:An 8-week feeding trial was conducted to investigate the effects of different lipid sources on the growth,serum biochemical indices and liver of juvenile yellowcheek carp (Elopichthys bambusa).Six iso-nitrogenous (49.43%±0.19%) and iso-energetic (18.09±0.03 MJ/kg) experimental diets were formulated,using 100% fish oil (3.80 g/100 g diet) as the control diet.Half of the fish oil of the other diets were replaced with different lipids:corn oil (CO),sunflower seed oil (SFO),palm oil (PO),soybean oil (SO) and pork lard (PL),respectively.Juvenile yellowcheek carp,with initial body-length of 6.89±0.20 cm and initial body-weight of 3.09±0.23 g,were cultured in 18 cylindrical tanks (340 L in volume,containing 280 L water) with 15 individuals in each tank.Water was aerated and circulated with a daily exchange rate of 15%-20%.The juveniles were fed twice (08:00 am and 16:00 pm) daily.During feeding trial,water temperature was fluctuated around 25-28℃ and pH value was 6.5-7.5.The final body-weight and weight growth rate of the juveniles showed a order of SFO> FO> SO>PO>PL>CO,with no significant difference among SFO,FO,SO and PO,but significantly lower in groups PL and CO.No significant difference of protein efficiency ratio(PER) and feed conversion rate were observed among groups.Alanine transaminase (ALT) from group CO was significantly higher than the control group; whereas aspartate transaminease（AST）was highest in groups CO and PL.Fat droplets in liver cells from group PL was obvious compared to other groups.Results indicated thatsunflower seed oil，palm oil and soybean oil are supposed to be suitable dietary lipid sources for juvenile yellowcheek carp.

Abstract:The proximate nutritional composition and odor components of the silver carp bone soup cooked under high pressure and atmospheric pressure processing condition were determined to provide the fish bones soup processing method.The results indicated that the contents of total solids,protein,water-soluble protein,calcium,ash and free amino acid under high pressure were significantly higher than that of atmospheric pressure except arginine and proline.The content of monounsaturated fatty acid under high pressure were 14.16% higher than that of atmospheric pressure.The odor components were extracted and determined by headspace solid phase microextraction and gas chromatography-mass spectrometry,63 volatile components were identified from the soup under high pressure while 25 volatile components under atmospheric pressure.To obtain the maximum nutrition value and best flavor,the fish bone soup should be cooked under high pressure.The temperature should be controlled between 110℃ and 120℃.The ratio of bone to water should be between 1∶1 and 1∶1.5 and the soup should be simmered between 1.5-3 h.

Abstract:Threshing characteristic and analysis of full ripeness period rape

Abstract:Postharvest diseases caused considerable losses to harvested fruits during the process of postharvest storage, transportation and distribution. By now,postharvest pathogens of fruits were controlled primarily by chemical fungicides. Therefore,it is urgent to develop alternative methods because of the fungicide residual,the development of fungicide-resistant strains of pathogen, the environmental pollution and the threat on human health. As one of the new and safe methods for postharvest diseases of fruits,various antagonistic yeasts have been isolated and identified as the potential antifungal biocontrol agents against postharvest fungal pathogen in the past thirty years,some yeasts-based products have obtained the permission to be used commercially. In recent years,mechanism of action of antagonistic yeast involved in the biocontrol of postharvest pathogens has been researched extensively,including the yeast′s competition for nutrients and space,the production of antifungal compounds,the resistance of fruit tissue and mycoparasitism. In this paper,the above-mentioned mechanisms of yeasts were reviewed,which will be helpful for selecting appropriate formulations and methods of application.