Abstract:218 field plots were sampled and characteristics of plant communities of the Sichuan snubnosed golden monkey habitat with various levels of degradation in Shennongjia district were compared. Community restoration models for degraded habitat were proposedusing community composition of the prime habitats. Results showed that the degree of habitat degradation in coniferous and coniferous and broadleaved mixed forests was higher than that in broadleaved deciduous forest.Tree species composition between prime habitat and degraded habitat was similar,but number of shrub species and food sources species in prime habitat was higher than that in degraded habitat. The main causes of habitat degradation were deforestation,insects and plant diseases,and overutilization by the golden monkey. Canopy density and average height of arbor decreased with the increaseof degradation severity. Eight different methods for habitat restoration for habitats underthelight,moderate and severe degree of degradation were put forward. It will provide direct scientific and practical support for habitat restoration and management of Sichuan snubnosed monkey in Shennongjia National Nature Reserve.

Abstract:To study the pathotypes of Magnaporthe oryzae in southwestern Hubei Province,seven Chinese identification rice cultivars and 15 nearisogenic lines of rice with single blast resistant gene were inoculated with 136 strains isolated from rice infected during 2012 to 2014,using Lijiangxintuanheigu(LTH) as susceptible control.The results showed that 121 strains with pathogenicity to LTH were belonged to 13 physiological races in 6 groups including ZA,ZB,ZC,ZE,ZF,and ZG based on the pathogenicity to the seven Chinese identification rice cultivars.Among these groups,ZB was the dominant population,while ZC increased significantly over the years,reaching about the same frequency as ZB in 2014.ZB13 was the first dominant physiological races,while ZC15 and ZB15 increased significantly over the years.According to the phenotypes of the 15 nearisogenic lines after inoculation,the 121 strains were divided into 77 pathogenic types,among which strains isolated from different years exhibited large difference in virulence.Over 90% of the strains tested were virulence tomonogenic rice lines with Pib,Pit,and Pi11,while less than 10% were virulence tomonogenic rice lines with Pi9 and Pikh.It will provide guidelines for local farmers,breeders,and agricultural sectors in choosing and developing rice varieties in this region.

Abstract:To study proteome of Rhizoctonia solani AG1 IA, the causal agent of rice sheath blight disease, three protein extraction methods (TCA-acetone precipitation method, SDS method and TCA-acetone-phenol/SDS method) were compared and a two-dimensional electrophoresis procedure was set up. Gel electrophoresis patterns were analyzed by Quantity One software and the results showed that 557, 309 and 877 protein spots were calculated by using TCA-acetone precipitation method, SDS method and TCA-acetonephenol/SDS method, respectively. Protein extracted by SDS method contained impurities such as salt ions and small molecules, and exhibited the fewest protein spots. Protein obtained by TCA-acetone precipitation method was yellow, indicating pigments contamination. Finally, a combined TCA-acetone precipitation and SDS method could get rid of salt ions, small molecules and pigments and produce a clear and high resolution electrophoresis pattern. Therefore, an optimized TCA-acetone-phenol/SDS extraction method was established in this study, which was suitable for proteins extraction from R. solani AG-1 IA for two-dimensional electrophoresis.

Abstract:Soil and root samples were collected from the rhizosphere of crops during the survey of plant nematodes in the main cultivation area in Tibet.Plant nematodes were isolated from these samples by using the modified Baermann funnel method.Based on morphological characteristics,three species,Pratylenchus loosi,P. neglectus and P. scribneri from genus Pratylenchus were identified.The P. loosi populations from Tibet has a shorter tail than those described in literatures.The main morphological characteristics and measurements of the P. neglectus and P. scribneri populations in Tibet were similar to those described in literatures.The P. neglectus was distributed in Nyingchi and Lhasa area,and the other two species were distributed in Nyingchi.

Abstract:High-throughput sequencing was used to study the transcriptomic changes of wildtype arabidopsis（Col-0）after salicylic acid treatment.12 novel genes downregulated by salicylic acid were identified. Among them,NG314 was studied and cloned from the cDNA of Col-0 seedlings. The total length of NG314cDNA sequence is 1 097 bp. The genomic DNA of NG314 contains one intron. Results of fluorescence quantitative PCR analyses showed that the expression of NG314 gradually decreased under the salicylic acid treatment.Pathogen infection repressed the expression of NG314. It is indicated that NG314 may encode a long noncoding RNA or novel miRNA,and may regulate plant disease resistance through repressing the expression of its target genes.

Abstract:The aim of the present study was to find some important metabolic biomarkers of Arabidopsis thaliana roots infected with Plasmodiophora brassicae at the late infection stage. P. brassicae spores were extracted from callus cultures of clubroot gall sections, cocultured with A. thaliana seeds on 1/2 MS medium and then after 45 d metabolic changes of the A. thaliana roots were analyzed by liquid chromatographymass spectrometry (LC-MS) technique. The results showed that P. brassicae could grow and develop in callus, and after cocultured with A. thaliana for 45 d, microscopic observation showed that the pathogens had a complete life cycle on the medium. Multivariate statistical methods of PCA and PLSDA suggested that the metabolities of experimental group (inoculated with P. brassicae) and control group (uninoculated with P. brassicae) differentially expressed. The results of OPLS-DA and t-test showed that the metabolites of flavonoids, lignans, terpenoids, steroidal glycosides, brassinosteroids and phospholipids were significantly increased during P. brassicae infection, suggesting their potential important roles in interaction between A. thaliana and P. brassicae at the late stage of infection.

Abstract:Carbohydrate active enzymes protein (CAZymes),one large kind of important proteins of plant pathogenic fungi and bacteria,plays an important role in its growth and development. Trichoderma virens Gv29-8 is the important bio-control agent of Trichoderma ssp.,inhibiting many plant pathogens. Predicting and analyzing the characteristics of the CAZymes is of great significance. Based on 377 secreted proteins obtained preliminary data,CAZymes protein was predicted with CAT forecasting procedures. 185 CAZymes proteins were found in T. virens Gv29-8 and divided into major categories and composite category. The former includes 71 GHs,54 CBMs,12 AAs,25 CEs,3 PLs and 3 GTs; the latter includes 17 CAZymes proteins. Genetic analyses showed that CAZymes classification should be further refined to a smaller category. It will lay a solid theoretical foundation for further studying its function.

Abstract:Combined with hairy root transformation,CRISPR/Cas9 is used to edit multiple sites in Nod factor receptor 1(NFR1) of Lotus japonicus and Glycine max, to quickly obtain the target mutants.The results showed that target gene mutants were generated in both of them. Editing efficiency of target 1 in LjNFR1 was 58.3%.No mutation was detected on target 2.The efficiency of target 1 and target 2 in Glycine max was 33.3% and 41.7%,respectively.Further sequencing analysis demonstrated that the CRISPR/Cas9 system is capable to simultaneously knockout multiple homologous genes in Glycine max,implying that the CRISPR/Cas9 can functionally manipulate redundant genes in plants.Exertion of the CRISPR/Cas9 system in transgenetic hairy roots shortened the experimental cycle of obtaining the desired mutants.

Abstract:The dynamic changes of bacterial community in two eutrophic areas including Guanqiao Lake and Sea World,referred to as H and L,of Donghu Lake in spring and summer were studied with highthroughput sequencing. The results showed that the bacterial community structures in H and L areas were similar.Actinobacteria were dominated in these area,following by Cyanobacteria and Proteobacteria. The relative bacterial abundance varied with time in both areas tested. A large fluctuation was observed in H area that Cyanobacteria (10.2%-33.7%) and Actinobacteria (15.4%-53.1%) abundance varied with significantly negative correlation,while bacteria were relatively stable in L area at the phylum level. The abundance of several classes including Actinobacteria,Synechococcophycideae,Oscillatoriophycideae and Chloroplast in H area,and the Oscillatoriophycideae and Chloroplast in L area fluctuated greatly. The diversity index of L area was higher than that of H area. Results of RDA analyses showed that pH,water temperature and dissolved oxygen were significantly negatively correlated with bacterial community in both areas,while N/P and TP were strongly positively correlated with bacterial community of H and L areas.

Abstract:The partial cDNA sequences of MHC I genes were obtained from grass carp (Ctenopharyngodon idella) genome sequence based on local Blast against zebrafish's MHC I region. Gene expression levels of BRD2, KNSL2, RXRB, TAPBP, FABGL, MHC Ia, PSMB9 and TAP2 in various tissues were detected by semi-quantitative RT-PCR. The results showed that expression of 7 genes above were highest in the blood but that of FABGL was highest in the kidney. After Aeromonas hydrophila infection at 4, 12, 24, 48 and 96 h, expression level of immune-related MHC I genes was detected in the spleen and kidney with quantitative realtime PCR(qRT-PCR). The expression level of BRD2 gene reached the peak at 12 h in the kidney but at 24 h in the spleen. The expression level of MHC Ia gene increased first and then decreased both in the spleen and kidney, reaching the maximum at 24 h. In the spleen, the expression of PSMB9 increased signicantly and reached the peak at 12 h, and then decreased signicantly at 48 h, following by a recovery to normal level at 96 h; in the kidney, the expression peaked at 24 h, and then continued to decline. The expression levels of TAP2 gene were first down regulated and then up regulated in the spleen and kidney, and backed to normal level at last. It is indicated that MHC I genes are related to the immune of grass carp, laying a foundation for further studying fish diseases and breeding resistance cultivars.

Abstract:To investigate the adherence of Bacillus subtilis and its inhibitory of Aeromonas hydrophila, the adherence rate of B. subtilis, its inhibition rate of A. hydrophila, and various physiological and biochemical indexes were detected in the intestinal epithelial cells of grass carp. The results showed that after treatment of B. subtilis alone, cell morphology, methyl thiazolyl tetrazolium optical density (MTT OD) value, lactate dehydrogenase (LDH), alkaline phosphatase (AKP) and glutamic oxalacetic transaminase (GOT) activity were unsignificantly changed (P>0.05), but the activities of Na+, K+-ATPase and glutamicpyruvic transaminase (GPT) significantly increased (P<0.05) at 3 h and 6 h, respectively. The cell shape was obviously changed by A. hydrophila from ellipse to irregular shape and the number of dead cells in the culture medium increased. Meanwhile，A. hydrophila could significantly increase MTT OD value and LDH activity in the culture medium, and significantly decrease the activities of Na+,K+-ATPase, GOP and GPT in the cells (P<0.05). The adherence experiment showed that B. subtilis could significantly reduce the adhesion of A. hydrophila to grass carp intestinal cells. In brief, adhesion and damage of A. hydrophila to intestinal epithelial cells were inhibited by B. subtilis.

Abstract:A probiotic strain dlt7a was isolated from feces of the Shennongjia golden snubnosed monkey and was identified as Enterococcus faecalis.In order to improve the viable count of dlt7a in the fermentation broth,the fermentation technology of dlt7a was optimized.The optimal culture condition with 20% (50 mL/250 mL) loading volume,2% inoculation amount,and initial pH of 6.5,was obtained by single-factorexperiment.In the study,Plackett-Burman experiment and response surface experiment were designed to identify the main nutrients affecting the viable count of dlt7a in the fermentation broth and to optimize the content of the three main nutrients including sucrose,fish meal and KH2PO4.The optimum nutrients in the media consisted of 2.55% sucrose,2.63% fish meal,2.5% soya bean,0.4% yeast extract,1.0% CaCO3,0.1% KCl,0.1% MgSO4·7H2O and 0.014% KH2PO4.E. faecalis dlt7a were then cultured by the optimized fermentation medium and condition,and the viable count reached 58.83×108 cfu/mL,which was 46.53% higher than that not optimized.

Abstract:The physical and chemical properties and bioactivities of the two kinds of SDF from black and white glutinous rice bran were studied to explore the possible role of SDF in lowering the blood glucose levels after the meal by digesting and absorbing glucose in vitro.The results showed that two samples had the typical characteristic of polysaccharide based on analyzing the infrared and ultraviolet spectrum.The contents of phenol,viscosity,molecular weight,assembly of aggregated particles,porosity of surface and pore size,oxidation resistance of SDF from black glutinous rice bran was favorable for absorbing glucose and inhibiting glucose diffusion,better than that of SDF from white glutinous rice bran.But the dissolution rate,the ability of inhibiting starch digestion and α-amylase activity of SDF from white glutinous rice bran was higher than that of SDF from black glutinous rice bran.It is indicated that the nutritional value and bioactivity of SDF from black glutinous rice bran is better than that from white glutinous rice bran.

Abstract:A set of hydraulic drive system with series-parallel combined double pumps and motors was designed to solve the problems of many moving parts and a long mechanical power transmission line of traditional rapeseed combine harvester. The system was applied in 4LYZ-1.8 rapeseed combine harvester. The relationship between throttle position and rotate speed was determined with testing hydraulic system. Orthogonal experiments were performed to study effects of header speed,cylinder speed,throwing machine speed and the speed of forced feeding roller on the power consumption of load sensitive system. The power consumption of circuits with main components was measured based on the experiment of power analysis. Results of analyzing variance of orthogonal experiments showed that power consumption of load sensitive system was significantly sensitive to the speed of threshing cylinder. The speed of cyclone separation tube inlet and the speed of header significantly affected the power consumption. Results of analyzing power consumption of hydraulic circuit showed that when the average feed rate was 1.5 kg/s,the average power consumption of five concerned parts including header,forcible feed rotor,threshing cylinder,throwing machine (conveying appliance was belonged) and fan was 1.68 kW,1.00 kW,5.11 kW 2.28 kW and 1.80 kW,respectively. Results of the field experiment of rapeseed combine harvester conducted in Huazhong Agricultural University indicated that the hydraulic drive system constructed can meet the working demand of rapeseed combine harvester,realize the regulation of stepless speed,and adapt different operation conditions. This study can simplify the structure of transmission system for rapeseed combine harvester and offer the reference for the power distribution of different components.

Abstract:A knurled cylinder for pneumatic cylinder-type centralized precision metering device for rapeseed was developed.The mechanical model of the single rapeseed on the compulsive driving layer in the filling area was established via analyzing the motion trajectory of each single seed.The EDEM simulation was conducted to study the effects of the knurled structure,cylinder rotational speed and seed filling height on the dynamic characteristics of seed.The results showed that the seed filling angle and filling performance increased with the increase of the friction between the cylinder and rapeseed.The cylinder rotational speed and seeding filling height affected the angle of seed filling and towing.With the increase of the cylinder rotational speed,the normal force and tangential force between cylinder and rapeseed,the total force and velocity of the rapeseed increased linearly.With the increase of the seed filling height,the total force and velocity of rapeseed increased linearly.The dynamics characteristics of rapeseed with knurled cylinder was better than that of the normal cylinder.The results of bench experiments showed that the positive pressure was 500 Pa,the seed filling height was -5 mm,and the cylinder rotational speed was 30 r/min with good seedling performance when the negative pressure was about 2 500 Pa.When the quality index was 90.10%,the loss index was 1.56%.When the cylinder rotational speed was between 15 r/min and 50 r/min,the variation coefficient of the consistency of each line displacement,stability of the total displacement and rapeseed damage rate were less than 5.5%,2.0% and 0.2%,respectively.The results of the field experiments showed that the pneumatic cylindertype centralized precision metering device for rapeseed oil satisfied the technical requirements of rapeseed planter with which the knurled cylinder was the key part.It will provide a reference for optimizing the structure of centralized precision metering device and improving seedling performance.

Abstract:A kind of tracked harvester for ratoon rice with double-headers and doublethreshing cylinders was designed based on wide swath and light-weighting concept to reduce rolling rate of harvester after harvesting the main crop rice and to improve efficiency of ratoon rice. The harvester was made up of two headers,two axial threshing cylinders with spike-tooth,two cyclone separating and cleaning devices,two granaries and a tracked chassis. This kind of threshing cylinder had a good adaptability to ratoon rice. The cyclone separating and cleaning device had advantages of light mass and low-power consumption. The swath of this tracked harvester reached up to 2.55 m,with feeding quantity of 1.6 kg/s. The material was “Zhongxiang 1”. The working speed,rolling rate and impurity rate were selected as main evaluation indexes. The results of field performance test showed that the working speed,working efficiency and the range of cutting height was 0.24 m/s,0.133 hm2/h,0.35~0.55 m,respectively. This combine harvester had the advantages of light mass,lowrolling rate. It can meet the requirements of harvesting for ratoon rice.