Abstract:To better understand the molecular mechanism of rice (Oryza sativa L.) in response to cold stress and further improve its tolerance to cold stress,a cold-responsive gene OsLCL3 was screened from cold stress expression profiles of rice gene chips.OsLCL3 was confirmed to be induced by low temperature (4℃) strongly with real-time fluorescent quantitative PCR and the expression of GUS reporter gene expression in stable transgenic rice driven by the promoter of OsLCL3.Homologous protein sequence analysis showed that OsLCL3 and its homologous proteins belong to DUF761 super family.Phylogenetic analysis of part members of this family including four homologs in rice genome was conducted.Subcellular localization analysis of OsLCL3 in rice protoplast showed that it was localized in nucleus and cytoplasm.Transgenic rice lines with over-expressing OsLCL3 were obtained and three of them were tested for low temperature stress tolerance.Results showed that the electrical conductivity of overexpression lines under the low temperature stress was significantly lower than that of wild type plants.The survival rate of overexpression lines was significantly higher than that of wild type plants.These results indicate that OsLCL3 may play an important role in low temperature response in rice.

Abstract:The genetic linkage map was constructed by using one F2∶3 population of crossing BR96 with Bai56 and 122 molecular makers covering the whole rice genome.The quantitative trait loci (QTLs) for BPH resistance were identified based on marker geneotypes and the pheontypes of BPH resistance in rice seedlings.Totally three QTLs for BPH resistance were detected.QBph3 was detected in the long arm of chromosome 3 between the molecular markers RM489 and RM282,with a LOD value of 5.1 and explained 3.8% the phenotypic variation.QBph4 was detected in the chromosome 4 between the molecular markers RM16605 and RM16717,with a LOD value of 28.7 and explained 29.4% the phenotypic variation.QBph6 was detected in the short arm of chromosome 6 between the molecular markers RM276 and RM527,with a LOD value of 2.7 and explained 7.1% the phenotypic variation.Obviously,QBph4 with the largest effect among three identified QTLs may be a major QTL for BPH.

Abstract:The conventional and hybrid japonica rice cultivars in Liaoning Province with three panicle types and three row distances and two hill distances were used to study the effects of physic and mechanic features,panicle factors under various row-hill spacing on the lodging resistance of japonica rice cultivars.The results showed that row and hill distances simultaneously affected the physic and mechanic features of japonica rice plant,thereby making japonica rice with different panicles exhibit different lodging capacity.Liaoyou 52210 significantly increased plant height and decreased flexural strength from 20 cm hill distance,which is the main factors of higher lodging index.Traits including plant height,the length and fresh weight of base N2 section,panicle angle,panicle length,fresh panicle weight and panicle gravity of Shendao 47 and Shendao 6 at the configuration of 25 cm×20 cm decreased significantly.At the configuration of 35 cm×15 cm,the cross-sectional area and flexural strength of the three japonica rice cultivars were significantly greater than that of others.From the perspective of panicle type,the adaptabilities of lodging resistance of Shendao 47 and Shendao 6 to the change in the configuration were better than that of Liaoyou 52210.The panicle angle of three japonica rice cultivars at the configuration of 25 cm×15 cm,30 cm×20 cm and 35 cm×20 cm was significantly larger than that of others.The fresh panicle weight,panicle length and height of panicle gravity at the later two configurations were higher as well.Generally,potential lodging and less panicles of per unit area at the configuration of high and low density limited the obtainment of high and stable yields.The flexural strength and bending moment at the configuration of 25 cm×15 cm and 35 cm×20 cm was the inducible factor increasing lodging index,indicating that both tense and sparse planting are unfavorable for lodging resistance and the flexural strength of base internode is the main target to increase the lodging resistance.The suitable bending moment of base internode should be considered when selecting cultivars with lodging resistance because the greater biomass is the guarantee for lodging resistance.

Abstract:SSR(simple sequences repeat) and SRAP(sequence-related amplified polymorphism) markers were used to analyze the genetic diversity and relationship of 34 citrus germplasms in Guangxi Province including 11 wild citrus ，13 local varieties and 10 control varieties.Results showed that 22 primer pairs of SSR were selected to obtain 221 polymorphic bands with 10.1 polymorphic bands per primer pairs.11 primer pairs of SRAP were screened to obtain 215 polymorphic bands with 19.5 polymorphic bands per primer pairs.The genetic similarity of SSR and SRAP for all germplasms accessions was ranged from 0.78 to 0.96 and 0.60 to 0.92,respectively.Both SSR and SRAP can efficiently identify real citrus including the lime and lemon,the pummelo,the mandarin and assorted hybrids,the Citrus ichangensis Swingle,and the wide leather citrus.

Abstract:In order to verify the effects of soil physi-chemical factors on the banana fusarium wilt pathogen (Fusarium oxysporum f. sp. cubense race 4,Foc 4),various soil physi-chemical factors including soil types,temperature,water content and pH value were simulated under laboratory conditions,and the effects on the growth and infection of Foc 4 were observed and analyzed.The results showed that the optimal growth conditions for Foc 4 were at 25℃ with higher soil moisture of 30% and acidic pH value of 4 to 5 in soil; whereas the optimal infection conditions of Foc 4 to banana plantlets were at 30℃ and in the acidic soil environment (pH 3 to 5) in the pot experiments.

Abstract:Lower the alternating wet and dry conditions with pH 5.0,the mixed solutions of FeSO4 and MnCl2 with different concentrations (0.00-0.08 mol/L) were added to yellow-brown soil columns with soil profile layer B leaching 60 times from the Xiaogan City,Hubei Province.The leaching and deposition of iron-manganese oxides of different forms and its adsorption and desorption characteristics on Pb2+ were investigated.The results showed that the manganese oxide content of the soil surface showed an increasing trend with the increase of leaching solution of manganese concentration，and the iron oxide content did not change obviously when the concentration of Fe2+ in the leaching solution was 0.04 mol/L and 0.08 mol/L.The deposition amount of free and amorphous iron oxides indicators was higher than that of manganese oxides in the soil surface，mostly upper layer (0-10 cm)>lower layer (20-30 cm) lower the same treatment.The adsorption quantities of Pb2+ in the soil samples after leaching 60 times increased with the increase of equilibrium concentration.The maximum of adsorption in upper layer and lower layer was 167 mmol/kg and 195 mmol/kg,respectively.The adsorption quantities of Pb2+ in the yellow-brown soil samples increased with the increase of the amount of manganese deposition.However，the desorption of Pb2+ showed the opposite trend.The trend of Pb2+ desorption quantities increased with the increase of Pb2+ adsorption quantities.

Abstract:To obtain small molecular antibacterial compound and its biosynthetic gene cluster,a genomic cosmid library was screened by high-throughput conjugation,heterologous expression,bioassay,and combined with DNA sequencing.A positive clone 1H5 expressing growth-inhibition activities against gram positive bacteria and Rhodotorula was obtained from the genomic library of Streptomyces hygrospinosus.The cosmid 1H5 harbored a cluster of genes highly similar to previously reported streptothricin biosynthetic gene clusters in terms of sequence and gene organization.Streptothricins were detected in the fermentation broth of Streptomyces expression hosts carrying 1H5,indicating that this clone contained an entire gene cluster for streptothricin biosynthesis.The cloning and heterologous expression of the biosynthetic gene cluster pave the way for further study of the biosynthesis,regulation and structural modification of streptothricin.

Abstract:The nodule-specific expressed protein AsF259 which are homologous to the CCPs was identified in Astragalus sinicus,expressed in Escherichia coli (DH10B) to study the subcellular localization in onion epidermal cell and in Astragalus sinicus nodule. The real time qPCR was used to analyze the gene expression. Results showed that AsF259 protein was distributed in the cell wall,cell membrane and cytoplasm,and highly nodule-specific expressed in the nodules with the highest expression level during the crucial stage of bacteroids differentiation. AsF259 protein was co-localized with the bacteroids,indicating that AsF259 gene belongs to a novel member of NCR family and may play an important role in bacteroids differentiation,maintenance and nitrogen fixation.

Abstract:One plasmid DNA with egfp and Kanr was introduced into the natural soil.The positive transformants were isolated directly from the bacterial suspension with flow cytometer and sorted with green fluorescence and kanamycin resistance.The frequency of natural transformation was less than 10-7 in soil.After the enrich cultivation,82 cells out of 39 736 426 were sorted.Positive transformants were finally obtained.The occurrence of horizontal gene transfer through natural transformation in soil was confirmed.Our studies break the lowest proportion limit of sorting with flow cytometry as well.

Abstract:Genetic diversity of 8 wild stocks of red swamp crayfish (Procambarus clarkii) from Tonglu (TL),Taihu Lake (TH),Hubei (HB),Jiangxi (JX),Hongze Lake (HZH)，Chongming (CM),Guangxi (GX),and Haiyan (HY),were investigated using 10 microsatellite DNA loci.In these 8 populations，the observed heterozygosity (Ho)，expected heterozygosity (He),and polymorphism information content (PIC) of these populations ranged from 0.525 7 to 0.740 0,from 0.623 8 to 0.743 4,and from 0.420 9 to 0.684 0,respectively,indicating a high level of genetic diversity.The highest and lowest genetic diversity were found in the Guangxi population and Taihu population,respectively.Then,genetic distance was calculated using the MEGA 4.0 software.And the genetic distance between HB population and TH population was the smallest,while that between CM population and JX population was the largest.

Abstract:The removal capacity of nitrogen and phosphorus by Ipomoea aquatica Forsk planted on artificial floating beds were emphatically studied,and the microflora structure in the yellow catfish (Pseudobagrus fulvidraco) intestine were analyzed by using high-through sequencing technique.The results showed that ammonia nitrogen,total nitrogen and total phosphorus can be removed by the I. aquatica Forsk planted on artificial floating beds,with high removal efficiency.There were some differences in the intestinal bacteria diversity between the experimental (with artificial floating beds) and control (without artificial floating beds) groups.But the bacterial composition of different groups did not significantly vary from each other,and the predominant species were similar in all groups.At the beginning period,Proteobacteria was dominant (88.56%) in the yellow catfish intestine,and at the ending period,the dominant bacterial community in the control group were Firmicutes (48.4%),Fusobacteria (32.52%) and Proteobacteria (14.56%),whereas in the experimental group,the core microbiota composed of Proteobacteria (36.72%),Firmicutes (21.89%),Fusobacteria (17.48%) and Bacteroidetes (11.48%).This study provided a more comprehensive understanding that I. aquatica Forsk planted on artificial floating beds can change the flora structure of the yellow catfish intestine.

Abstract:Eight hundred sixty eight Hy-Line Brown laying hens about 280 d old with similar egg production rate,were randomly divided into high temperature group (33℃),probiotics group (Bacillus subtilis+33℃) and room temperature group (26℃).The experiment was conducted for 20 d to investigate the feasibility and mechanism of B. subtilis HDRaBS1 on alleviating animal heat stress.The results showed that the egg production rate,average daily feed intake and average egg weight of the probiotics group increased by 2.60% (P<0.05),2.91% (P>0.05) and 0.96% (P>0.05),respectively,compared with the high temperature group,but were less than those of the room temperature group.In addition,the diarrhea rate and broken egg ratio of the probiotics group were reduced,serum endotoxin and IL-1were reduced by 37.41% (P>0.05) and 20.98% (P<0.05),while IL-10 expression was increased by 41.05% (P>0.05) than those of the high temperature group.Moreover,in the probiotics group,the mRNA expression of many genes in the cecum,including the tight junction occludin,ZO-1 and JAM-A,and the mucosa repair gene of EGF were significantly up-regulated (P<0.05),intestinal villus height (P<0.05) was improved,while no pronounced variation of the crypt depth was observed (P>0.05).Therefore,adding B.subtilis HDRaBS1 could effectively alleviate the symptoms of heat stress-induced laying hens.

Abstract:In this report, we isolated and identified a strain of Flavobacterium columnare (named Pf1 strain) from a diseased yellow Chinese catfish, Pelteobagrus fulvidraco. Subsequently, the drug sensitive tests and the pathogenicity of Pf1 on yellow Chinese catfish and mandarin fish (Siniperca chuatsi) were carried out and the results showed that Pf1 was pathogenic. Pf1 infection could cause necrosis and inflammation of several tissues, especially the liver, trunk kidney and gill in mandarin fish. In the infected mandarin fish, liver cells were swollen, vacuolar and necrotic. Necrosis of renal tubular and extensive inflammatory cells infiltration were observed. Gill lamellae capillaries were swollen and hyperemic, and respiratory epithelia were swollen and degenerated. The lesioned gill lead to the higher breathing rate, slower swimming, and eventually mass mortality of the infected mandarin fish.

Abstract:To select strains capable of degrading aflatoxin B1 (AFB1) efficiently, 32 bacteria strains were isolated from different sources which display AFB1 reduction activity, using coumarin as the sole carbon source and biodegradation technology. AFB1 degradation effectiveness was detected by high-performance liquid chromatography (HPLC) after AFB1 standard was added (final concentration of culture supernatant was 2.5 μg/mL). The bacteria strain isolated from chicken manure was named as F6, which could degrade AFB1 by 83.3%. Based on the morphologic, physiological, biochemical characteristics, the isolated strain F6 was preliminary identified as Bacillus. The analysis of 16S rRNA gene sequence homology revealed that, the nucleotide identity between strain F6 and Bacillus oleronius international standard strain ATCC700005 (accession number NR043325.1) was 99.3%. Thus, strain F6 was determined as B. oleronius and named as B. oleronius GX01 (Accession No. KP297896). The components of the fermentation broth of F6 were then isolated and detected. The results showed that the culture supernatant could degrade 83% AFB1, while the degradation rates of bacterial suspension and intracellular were 22.8% and 17.4%, respectively, suggesting that the degradation activity of B. oleronius strain F6 was exoenzyme.

Abstract:The research was conducted to construct an infectious clone of porcine circovirus type 2 recombinant virus ZJ-R. One copy and two copies in tandem of the whole genomic sequence of ZJ-R were cloned into the pSK vector with restriction site of enzyme EcoRⅤ. The B cell epitopes of the ZJ-R virus structural protein were predicted by using biotic soft-wares. The epitope peptides were chosen, synthesized and coupled to KLH. Then the New Zealand White Rabbits were immunized to elicite the polyclonal antibodies, which were used later in immunohistochemical techniques. The results showed that the ZJ-R virus could be detected by IHC after 72 h post transfection of pSK-2 ZJ-R. In conclusion, an infectious DNA clone of the porcine circovirus type 2 recombinant virus ZJ-R was constructed successfully.

Abstract:Fresh-cut lotus roots were used to study the effects of ultra high hydrostatic treatments on the quality of lotus roots during storage.With the increase of pressure and holding time of extending,soluble solid and total phenol had little changes.The content of ascorbic acid,the activity of PPO and the frangibility of lotus roots decreased significantly (P<0.05).The high hydrostatic pressure treatment significantly inhibited the microbial growth (P<0.05).The sterilization effect was not increased with the prolonging of holding time,indicating that the instant pressurization and instantaneous unloading process played an important role in ultra high pressure sterilization.Further studies of ultra high pressure treatment on fresh-cut lotus root showed that the loss of Vc under UHP treatment was lower than that of the control but the soluble solids loss increased gradually during storage.The color of lotus root changed little.The high hydrostatic pressure treatment inhibited the PPO activity and the microbial growth.It was clearly demonstrated under the microscope that the high pressure damaged the cell structure of lotus root.

Abstract:Yellow light was used to irradiate plucked leaves during its withering.The withered leaves were subjected to cutting and drying.The qualities of black tea were analyzed.The results showed that yellow light significantly improved the aroma and taste.The sweet flavor and fresh and mellow taste of black tea was less affected.The concentrates of catchins,theaflavins,amino acids and aroma components except polyphenols and soluble sugars were significantly increased as well.After irradiation of yellow light,the total content of catechins in black tea increased by 21.6%,among which EGCG,ECG,GC and EC were increased remarkably with no significant difference in EGC and C.The total content of theaflavins in black tea withered with yellow light had an increase of 17.6%.But each theaflavin showed different variation with a significant increment in TF1 and TF2A,but there was no significant difference in TF3 and TF2B.It is indicated that the amimo acids of black tea withered with yellow light increased by 6.8% with a significant increment in aspartic acid,glutamic acid,yaline,isoleucine and theanine.The result of GC/MS analysis showed that there was 21.7% increase of total content of aroma components.The predominant types of acids and anhydrides,alcohols,esters and aromatic compounds increased by 58.5%,41.2%,25.4% and 18.6%,respectively.Amides decreased by 127.3%.It is indicated that yellow light irradiation during tea leaves withering can improve the qualities of black tea.

Abstract:288 rice straw samples collected from different regions and varieties were used to study the feasibility of rapidly detecting the content of soluble sugar in rice straw with the near infrared reflectance spectroscopy (NIRS) technique.The near infrared spectral information of samples were collected within the near infrared wavelength range (10 000-4 000 cm-1).The models of quantitative analysis based on near infrared spectra of stepwise multiple linear regression (SMLR),partial least squares regression (PLS) and principal component regression (PCR) were established using stoichiometric algorithm SMLR，PLS and PCR，combined with different spectral pretreatments including multiplicative scatter correction (MSC),standard normal variation transformation (SNV),derivative,S-G smoothing and their combinations.Through comparison and analysis, the optimal effect of PLS model which established by using first derivative spectra pretreatment had a determination coefficient R2C between the calibration set chemical analysis values and predicted values of 0.880 6， the determination coefficient R2CV and R2V of 0.771 1,0.857 8, and root mean square difference RMSEC, RMSECV, RMSEP of 0.318%，0.440%，0.404%, respectively. Both of the relative analysis error RPDC and RPDV are greater than 2.5. The results show that establishing the model by using near infrared spectroscopy, combined with PLS modeling method can quickly detect the content of soluble sugar in rice straw.

Abstract:A system of online weight detection and weight classification for freshwater fish was designed.The system detected the fish weight with an electronic weighing module.Weight signal of fish body was collected by the pressure sensors when the fish reached weighting area.The signal was sent to the control chip for storage and analysis after processed by the conditioning circuit.After figuring out the weight of the fish,the control chip judged the weight grade of the fish and sent the grade information to the fish distribution mechanism for distribution.The photoelectric sensors sent location information to the control chip after the fish reached the sensors installed on the fish conveyor.The controlling system allocated corresponding mechanisms to finish distribution depending on the weight grade and location information.Different weight sets were used to test the accuracy of the weighting device.The results showed that the weighting error was between ±0.73% when the weighting speed was between 0.16 m/s and 0.24 m/s.Results of performance test showed that the system can process 26 fish per minute with the classification accuracy of more than 98%.The weighing speed was 0.20 m/s.The distribution speed was between 0.20 m/s to 0.25 m/s.The fishes weighed between 300 g to 3 000 g were divided into 4 classes.

Abstract:To reduce the cultivation process of heavy plow,leakage phenomenon of tillage,improve work efficiency,a method of identifying path for rotary cultivator based on the paddy field unstructured environment color image collected by visual navigation system was designed.Choosing the suitable color feature of paddy field was discussed.Considering that the soil area has less noise and the straw area has more noise,the over and compensation method was used to denoise the paddy image.The feature value of each pixel color was acquired by progressively scanning the image.The border information was obtained by using difference of the color feature between the straw area and soil area.The least squares technique was used to make a line and acquire the elevation angle which is the steering angle of rotavator.The rotavator's direction of advance which is the path information was determined.This method achieved good results in static image processing.It will be improved by the field test.

Abstract:An experimental system was established under different conditions of impacting materials,detected points,excitation points,impacting velocities,mass and eggshell strengths for eggs.The domain signal was received by the acceleration sensor and analyzed.Results showed that the dominant frequency was not significantly affected by the different impacting materials,detected points,excitation points,impacting velocities.It was the significantly affected by the mass and eggshell strength.The dominant frequency increased with the decrease of the mass,and increased with the increase of the strength of eggshell.