Abstract:One new record species of genus Lepiota is reported from China:Lepiota forquignonii Quél. Samples were collected from Jiamusi City,Heilongjiang Province. The new species is characterized by its ellipsoid basidiospores,widely clavate or obovid cheilocystidia, trichodermial palisade made up of differentiated elements,and clamp connections.It is placed in the genus Lepiota section Ovisporae.

Abstract:Lotus japonicus in legumes is one of the model plants in the study of the system on the symbiotic nitrogen fixation.The research of the symbiotic signaling pathway and the mechanism of interaction of rhizobial bacteria helps to discover the secret about the establishment of symbiotic relationship and the formation of the symbiont between rhizobial bacteria with legumes.In this study,the interaction of the two GRAS family proteins,nodulation signaling pathway1 (NSP1) and nodulation signaling pathway2(NSP2),was confirmed in vivo and vitro using a yeast two-hybrid system and protein-protein interaction in vitro.The interacting loci were assessed to be the LHRI domain of NSP2 and LHRII domain of NSP1.

Abstract:A β-mannanase producing strain CYS4 was isolated from factory waste liquid and its enzyme activity reached 11.0 U/mL in the culture liquid of shaking flask.CYS4 was identified as Bacillus subtilis based on morphological observation,physiological and biochemical experiments and phylogenetic analysis with 16S rDNA sequence.Enzymatic characterization showed that the optimum pH for the enzyme was 7.0,it was stable between pH 5.0 and pH 11.0,the enzyme activity is more than 80％,and the optimum temperature was 60 ℃,it was stable under 40 ℃,the enzyme was still more than 70％ after 3 hours.The enzyme activity was activated by Na+,Fe2+,Mn2+,Zn2+,Mg2+,but inhibited by Hg2+.

Abstract:Using rape and rice as explants,the colonization range,promotion of growth and aphid bioassay of the endophytic bacterium YY-23 and SJ-10 were tested in laboratory.The results showed that the two strains were able to colonize in rape,rice,wheat,maize,Chinese kale,navet,etc.Rape seeds were dipped for 24 h with YY-23 and SJ-10,then sowed.20 days later,the fresh weight of rape plants treated by YY-23 and SJ-10 was increased by 113.5% and 52.5%,respectively.The survival rate of aphid in SJ-10 group was lower than CK on the sixth day,the difference was significant (P<0.05).However,YY-23 expressed no resistance to aphid.Therefore,the two strains could stably colonize and promote growth of plant.SJ-10 had insecticidal effect on aphid.

Abstract:Fifteen pairs of primers were designed to detect single nucleotide polymorphisms of exon 1 to exon 9 of progesterone receptor (PGR) gene in Small Tail Han,Hu,Dorset and Texel sheep by PCR-SSCP.Only the products amplified by primer P9 displayed polymorphism.For primer P9,three genotypes (AA,AG and GG) were detected in Small Tail Han,Hu and Dorset sheep,and two genotypes (AA and AG) in Texel sheep.Sequencing revealed one mutation (217A→G) of PGR gene in the genotype GG in comparison to the genotype AA,and this mutation did not cause any amino acid changes.The ewes with genotype GG or AG had 0.97 (P<0.05) or 0.64 (P<0.05) lambs more than those with genotype AA,and the difference of the litter size between GG and AG genotypes was non-significant (P>0.05) in Small Tail Han sheep.These results preliminarily indicated that allele G of PGR gene is a potential DNA marker for improving litter size in sheep.

Abstract:In order to study the effects of different combination of regulating facts on growth performance,carcass traits and meat quality in broilers,630 one-day-old healthy avian broilers were randomly assigned into 21 groups (including one control treatment),the control treatment was fed by normal diet,and the rest 20 treatments were fed by fifteen kinds of experimental diets.Each of three experimental factors (compound vitamins,compound trace elements and compound active additives) was designed to 5 levels.After 49-days’ growth trail， the mean weight of broilers,the feed efficiency of broilers,breast muscle/live weight of broilers,and the abdominal fat rate,the pH value,the water holding capacity,intramuscular fat,crude protein of meat in each treatment were better than those of the control treatment (P＜0.01).The multi-parameters comprehensive evaluation and optimization of mathematical models showed that the carcass and meat quality of broiler were the best when three compound additives in the diet was 1 672.64 mg/kg compound vitamins （VE 254.91，VC 283.23，VA 4 248.51，VH 0.43），2 888.10 mg/kg compound trace elements(Fe 77.84，Cu 90.81，Mn 116.75，Cr 0.78，Zn 142.70，Se 0.193),and 1 885.01 mg/kg compound active additives(betaine 1 767.53,allitridum 81.40 ),respectively.These results indicated that the optimal ratio of basic nutritional factors and regulatory factors could improve carcass traits and the meat quality of broiler.

Abstract:Under the current production facilities equipment and technical available level in China,light engergy utilization efficiency and its main parameters influenced by light energy utilization efficiency of canopy cross section,canopy vertical section,photosynthetic organs and photosynthetic apparatus,were analyzed in greenhouse tomato,in order to exploit the solar energy utilization efficiency from the macro leaf canopy interception of light energy to micro original dynamics of light energy conversion and kinetics of photosynthetic synthase.The leaf light absorption coefficient(ε) of tomatoes in greenhouse was significantly different between genotypes and leaf position with red and blue light irradiation by using globale integral instrument.The leaf red and blue light absorption coefficient ratio(εred/εblue) of ‘Baoluota’ variety was more than 1,which indicated that leaf had strong absorption capacity to red light.However,the ratio of εred/εblue of ‘Harvest’,‘Laifu 60’,‘Hongli’ and ‘Israel 1498’ varieties was less than 1,which indicated that leaf of these varieties had strong absorption capacity to blue light.Increasing row space could inhibited effectively group unrestrained growth and decreased the leaf area index (ILA) by 22.3%,which could avoid closure of inside canopy.As the degree of leaf-cutting increasing,the tomato plant width in greenhouse significantly decreased,meanwhile the overlap rate of plant width(ξp),the overlap coefficient of leaf area(ξ) and complex light absorption coefficient (α) of canopy consisting with canopy leaves and canopy air space were all decreased accordingly.Using the techniques of cutting-half-leaf with the close-planting,namely,ear inferior leaves were cut 1/3-1/2 as fruit expanding stage to maintaining overlap rate of plant width(ξp) between 10% and 20% when the planting density increased by 17%-30%,achieved the double effects of both improving the light energy interception of middle and lower canopy and enhancing the groups photosynthetic capacity.

Abstract:Based on screening several key reaction conditions of ISSR (inter-simple sequence repeat) markers,such as the concentrations of template DNA,primers and Mg2+,an optimal ISSR reaction system for Chimonanthus nitenswas established,namely,40 ng genomic DNA,0.3 μmol/L primer,1.5 mmol/L Mg2+,1 U Taq,0.2 mmol/L dNTP,and 2.5 μL 10×buffer were contained in 25 μL reaction system.Using this optimal ISSR reaction system,12 polymorphic primers were screened out.Furthermore,the best annealing temperature of the every selected primer was decided according to the amplified results with different temperatures among 51 ℃ and 56 ℃.

Abstract:The effects of precursors,metabolic intermediate of theanine synthesis and the NO donor SNP （sodium nitroprusside）on the synthesis of theanine in tea callus were investigated by using HPLC (high performance liquid chromatography) to detect theanine contents,and an accumulation of theanine in the roots of tea plants with different cultivation time after addition of (NH4)2SO4 into cultivation liquid was also discussed.As a result,25 mmol/L ZtNH2·HCl (ethylamine hydrochloride) could increase the content of theanine in callus significantly,especially in the callus grown from the 6th day to the 12th day.The theanine concentration in callus which grown for 12 days with addition of 25 mmol/L ZtNH2·HCl reached 108.96 μmol/g in dry weight basis,7.72 times of that in the control; addition of glutamine,alanine or sodium glutamate into medium with low concentrations could also increase theanine in callus,but the increasing amount was not very obvious compared to that with addition of 25 mmol/L ZtNH2·HCl with the exception of 5 mmol/L glutamine,which could boost the theanine to 7 times of that in the control.Addition of SNP with concentration of 0.05 or 0.075 mmol/L could increase the content of theanine in callus,but the increase amount was low in general.With addition of (NH4)2SO4 to 100 mg/L into cultivation liquid,the theanine content in tea plants was 2.25 times of that in the control on the 3rd day of the cultivation.The materials obtained with different theanine contents in this study could be used as materials for discovery of the genes correlated with theanine metabolism using digital gene expression profile in the future.

Abstract:The phylogenetic trees of the insect chemosensory proteins (CSPs) gene from 59 sequences of CSPs in 23 species under 7 orders published in GeneBank were generated by MEGA 5.0. Complete comparisons of CSPs sequences were used to determine the rates of synonymous(Ks) and nonsynonymous(Kn) codon substitutions and the ratio of Kn/Ks was calculated to estimate of the selection pressure on CSPs genes in insects.The Kn/Ks ratio was far less than 1,suggesting that CSPs may be evolving under negative selection.By the site-specific model in PAML 4.4, 6 amino acid sites undergoing positive selection were detected,5 of them were in the important function regions.

Abstract:A new venom allergen-like proteins,named Dd-vap-2,was isolated from Ditylenchus destructor by RT-PCR and RACE. The full length cDNA of Dd-vap-2 consists of 1 150 bp, with a 912 bp ORF encoding a 303 amino acid protein (GenBank accession No. GU370352).The putative protein with the oretical molecular weight was 31.8 ku in weigh and its pI was 6.76. The sequence analysis indicated that this gene was classified as a member of cysteine-rich secretory protein, CRISP and has a 21 amino acids long signal sequence at N terminal. Phylogenic analysis suggested that Dd-vap-2 had a close homology to Globodera rostochiensis and Heterodera glycines.

Abstract:Eighty one Setosphaeria turcica isolates were collected from 13 corn planting areas of Yunnan Province during 2007 to 2009, which were identified by using two sets of the corn differentials containing single Ht resistance genes and their mating types were analyzed by using molecular markers. The results showed that both physiological races and mating types of the pathogen were very complicated in Yunnan Province. The isolates could be divided into 11 races including 0, 1, 2, 3, 12, 23, 2N, 3N, 12N, 23N and 123N, among which 123N could overcome all of the single dominant resistance genes of the differentials to S.turcica and accounted for 22.2%. Based on the molecular markers, the 81 isolates were divided into three mating types including A, a and Aa, and neuter, accounting for 49.4%, 12.3%, 29.6% and 8.6%, respectively. In addition, different races and mating types of S.turcica existed in the same area.

Abstract:The low-molecular-weight antibiotic substances of low molecular weight induced in rice leaves by salicylic acid(SA) treatment were studied. Plate method was used to detect the effect of the extract on spore germination and Gas chromatography-mass spectrometry(GC-MS) was used to analyse these low-molecular-weight antibiotic substances. The results showed that the induced substances can effectively inhibit spore germination in Magnaporthe grisea.GC-MS analysis also revealed that SA could significantly induce many kinds of low-molecular-weight compounds in rice leaves and that their content was increased as compared to the control group. SA could cause the production of momilactone A, momilactone B, eugenol, and chorismic acid evidently，SA could alter the levels of phenol, methyl salicylate, stigmasterol, β-sitosterol, undecane and methyl ester. These results provided direct evidence that the rice plant produces phytoalexins in response to SA treatment.

Abstract:Detection on the virulence of Beauveryia bassiana MZ041016 against Frankliniellao ccidentalis (Pergande)

Abstract:Herbicidal activities of 29 plant extracts were tested against Pistia stratiotes L. in laboratory by water culture method at 20-25 ℃ and RH 50%-60%. Results showed that 10 of them had obvious herbicidal activities by the indicators of leaf mortality and whole plant mortality and with reference to root response. They were Aralia armata (Wall.) Seem.,Mahonia bealei (Fort.) Carr., Hibiscus syriacus L., Coptis chinensis Franch, Rhus javanica Mill., Datura metel L., Aleurites fordii Hemsl., Dipsacus chinensis Batal, Zanthoxylum simulans Hance and Tripterygium wilfordii Hook. f..Among these 10 plants,Aralia armata(Wall.) Seem. was the best one. When its methanol extracts reached the concentration of 0.625 mg/mL, the corrected leaf mortality of Pistia stratiotes L. was 84.56% and the corrected whole plant mortality was 80.64%. Even at the low concentration of 0.313 mg/mL, the corrected leaf mortality of Pistia stratiotes L. was as high as 55.65% and the corrected whole plant mortality was 38.97%.

Abstract:To overcome the drawbacks of stricture representativeness of quadrat,great error,resampling,the mode of yield estimation based on field window was proposed.Based on mechanism of remote sensing on crops using administrative village as survey region,the field extracted from QuickBird image and TM image was used as main data.Combining with the methods of field survey,a preliminary discussion about key technology of remote sensing yield estimation based on field window from two aspects of sampling field and region of corn and peanut was made.This study was attempted to extract the crops on large scale,considering that it would have better results of crops using classification supported by field window.It also showed that there was a linear relationship and significant positive correlation between NDVI and yield per unit area.The method using the TM image supported by field window and combining with field survey is considered to be a suitable mode of yield estimation for the southern China with disconnected field and complex plantation type.

Abstract:Undisturbed soil monolith lysimeters were conducted to study daily nitrous oxide (N2O) emission and its relationship with soil temperature and water content in a continuous vegetable cropping system in Wuhan.Results showed that N2O daily emission increased with the increase of soil temperature and the peak emission occurred at the highest temperature when soil moisture was relatively stable.However,when soil moisture changed rapidly in one day,the peak emission of N2O occurred at appropriate soil moisture and relatively high soil temperature,meaning that both soil moisture and temperature affected the daily change of N2O emission in this vegetable system.Nitrogen fertilizer was found to be another important factor in affecting daily N2O emission.

Abstract:Soil adsorption is the key factor of reducing the efficacy of pesticides against soil pest insects.Control of the red imported fire ants with surfactant-enhanced remediation (SER) was investigated and effects of two surfactants SBDS and Triton X-100 on desorption of contact insecticides (phoxim) was tested in the soil system with a batch equilibration technique.As the concentration of SDBS or Triton X-100 increased,the adsorption rates of phoxim decreased.At 5 folds of critical micell concentration (CMC),the adsorption rates of phoxim at the test initial concentrations treated with SDBS ranged from 56.81% to 66.21%,which were obviously lower than those (75.92%-82.07%) of Triton X-100 treatment,indicating that desorption of SDBS to phoxim in soil was higher than that of Triton X-100.The adsorption of phoxim in soil was conformed to Freundlich’s isothermal adsorption curve,which showed that the higher amount addition of surfactants decreased the Kaf value.These results might favor the application of phoxim for the control of the red imported fire ants.

Abstract:The aim of the study is to establish a procedure on root protein extraction applied for proteomics analysis in Brassica napus seedlings.Three methods (TCA/acetone,phenol extraction and phenol extraction-methanol/ammonium acetate precipitation) for protein extraction were assessed with Brassica napus cultivar “Qingyou 10” root at seedling stage as experiment materials.The protein concentration was determined using Bradford protein assay with BSA as a standard,and the protein samples were separated by SDS-PAGE and two-dimensional electrophoresis (2-DE) in turn.Then,the 2-DE gel images were analyzed by PD quest,detecting 644,645,669 clear spots,respectively.Evaluating comprehensively the efficiency of protein extraction among the three methods and optimal 2-DE condition,modified phenol extraction,along with the 2-DE parameters,17 cm (pH 5-8) IPG strip,150 μg protein loading volume and 10% PAGE gel concentration,were chosen to obtain high reproducible and clear 2-DE gel image,which could satisfy the demand for protein isolation and identification in proteomics study.

Abstract:The species and vertical movement pattern of bacteria was studied in shrimp-cultured saline-alkaline pond in Tianjin.The results were as following:the amount of bacteria remained between 2.4×10 3 and 8.5×10 5 cfu/mL,the amount of Vibrio remained between 1.75×10 2and 2.3×10 5 cfu/mL,the amount of bacteria and Vibrio got to the peak in August and September,respectively.The amount of denitrifying bacteria remained between 3.4×10 4and 2.8×10 5 cfu/mL,the amount of sulfate-reducing bacteria remained between 2.9×10 2 and 7.5×10 4cfu/mL,and the amount of ammonifying bacterium remained between 1.2×10 2 and 7.0×10 4cfu/mL.There were 13 kinds of bacteria were examined in the shrimp-cultured pond.Vibrio,Alcaligenes and Corynebacterium were the mainly species in the earlier period,and Alcaligenes,Vibrio,Aeromonas and Enterobacteriaceae were the mainly species in the intermediate period,and Vibrion,Aeromonas and Cytophaga were the mainly species in the later period.The bacteria diversity index were highest in July and lowest in September.

Abstract:Five electronarcosis groups and one control group were designed to study effects of electronarcosis on haematological biochemistry of juvenile Siberian sturgeon Acipebser baerii.Results showed that total protein and albumin of sturgeon in the electronarcosis groups increased positively with voltage intensity.The values of total protein in the 35 V group were significantly higher than those in the 15 V and 20 V groups.The concentrations of high density lipoprotein cholesterin,low density lipoprotein cholesterin and triglycerides were the lowest in the 35 V group and was the highest in the 15 V group.These three indexes in the 35 V group were significantly higher than those in the control group and the 15 V group.There was significant difference between the 15 V group and the 35 V group.The value of glucose in the control group was 2.21 mmol/L.The values of glucose in each electronarcosis group were higher than those shown in the control group.Urea and total bilirubin were the lowest in the 15 V group and the highest in the 35 V group.The values of alanine aminotransferase,aspartate aminotransferase and lactate dehydrogenase were relatively steady.The value of alkaline phosphatase in the control group was 218.33 U/L.The value of alkaline phosphatase in the 35 V group was significantly higher than that in the control group and the 15 V group.The present study showed that electronarcosis was a good anaesthesia method.The appropriate voltage of electronarcosis in Siberian sturgeon was from 20 to 30 V.

Abstract:An intracellular copper/zinc superoxide dismutase (cpSOD) cDNA was cloned from the haemocytes of freshwater mussel Cristaria plicata using reverse transcription-polymerase chain reaction (RT-PCR) by degenerate primers.Further studies were conducted on cpSOD mRNA expression in various tissues of C.plicata by semi-quantitative PCR method.The results revealed that the cpSOD cDNA clone consists of 891 bp with an open reading frame of 468 bp and with a protein of 155 amino acids.The calculated molecular mass is 15.8 ku with an estimated pI of 5.8.Four conserved amino acids responsible for binding copper (H46,H48,H63 and H120),four conserved amino acids responsible for binding zinc (H63,H71,H80 and D83) and a conserved intramolecular disulfide bond were observed.Sequence comparison showed that the deduced amino acid sequence of cpSOD presented 70% similarity to that of saltwater shellfish Crassostrea gigas ic-Cu/Zn-SODand 69% to Chlamys farreri ic-Cu/Zn-SOD,respectively.Semi-quantitative RT-PCR analyses showed that cpSOD genes were expressed in haemocytes,mantle,gill,adductor muscle and hepatopancreas of C.plicata.Stimulating C.plicata by injection with Aeromonas hydrophila significantly up-regulated the expression of cpSOD in haemocytes,indicating that cpSOD is involved in the immune responses against A.hydrophila.

Abstract:Collagen was extracted from grass carp scale with acid-enzyme combination-method for preparing fish scale collagen film (FCF) by the plate pouring method.The film-forming conditions and chitosan on the properties of gelatin film were studied to optimize the preparation conditions of FCF.The results showed that the conditions forming scales and polysaccharides have significant effects on the fish scale collagen membrane characteristics.The optimum conditions were 45 ℃,pH 5,collagen/chitosan=6∶4.The tensile strength was 61.27 MPa,and elongation at break was 5.17%.

Abstract:Chloroplast transformation in plants has many advantages over nuclear transformation.Proteins in chloroplasts can be expressed at high levels with proper folding and disulfide bonds as the cells of higher plants contain a large number of chloroplast genomes.Multiple genes can be co-expressed in chloroplast genomes.Furthermore,chloroplast genes are inherited in a strictly maternal fashion in most angiosperm plant species,and this minimizes the possibility of out-crossing transgenes to related weeds or species.In addition,gene silencing,position effects and random integration have not been reported in chloroplast transformation.Although chloroplast transformation is very attractive,this technology is not as widely used as nuclear transformation.It has been mostly focused on 16 plants species,especially tobacco in which many proteins has been expressed including vaccines and antibodies.In this review we briefly summarize the rationales,methodologies,applications,bottlenecks and prospects of this promising genetic engineering technology for chloroplasts.