Abstract:In order to investigate the relationship between the tendon development related genes (tnmd/xirp2a) and the intermuscular bone in blunt snout bream (Megalobrama amblycephala),we firstly obtained the cDNA sequence of tnmd/xirp2a by cloning. The fulllength cDNA of tnmd is 1 420 bp,including an open reading frame (ORF) of 906 bp encoding 301 amino acids. The cDNA sequences of xirp2a is 11 715 bp in length,including an ORF of 9 522 bp encoding 3 174 amino acids. Phylogenetic analysis showed that the tnmd gene was lowly conserved among different vertebrates. The tnmd protein had a sequence deletion compared to mammals and poultry. The tnmd/xirp2a expression in the muscle and intermuscular bone was significantly higher than that in other tissues (P<0.05). The tnmd expression in the muscle was significantly higher than that in the intermuscular bone (P<0.05),while the result of xirp2a was the exact opposite. During intermuscular bone development,the tnmd mRNA was significantly higher at stage Ⅲ than that at the other three stages. However,there was no significant differences of the xirp2a expression among stages Ⅱ,Ⅲ and Ⅳ (P＞0.05). Our results indicate that the tnmd gene may play possible regulation roles during the intermuscular bone development,while the relationship between the xirp2a gene and the intermuscular bone development needs to be further studied. The results of this study provide theoretical support for the derivation of intermuscular bone from the tendon.

Abstract:This study was conducted to investigate the function of the SREBP1 gene inMegalobrama amblycephala. The SREBP1 gene was cloned by RTPCR,and its expression were analyzed using qRTPCR. Bioinformatics analysis showed that the cDNA coding sequence (CDS) of SREBP1 was 3 426 bp (GenBank Accession number:MH633449),with an open reading frame (ORF) encoding 1 141 amino acids. The M. amblycephala SREBP1 protein shared high similarity (74%~99%) with Salmo salar,Danio rerio,Ctenopharyngodon idella,Cyprinus carpio and Sinocyclocheilus grahami. Conserved domain analysis showed that the SREBP1 protein has the bHLHZip domains,which belong to the SREBPs family. Expression analysis showed that the SREBP1 expression leveled was the lowest at the optic vesicle stage and highest at the hatching stage during embryonic development of M. amblycephala. For juvenileM. amblycephala,the SREBP1 gene expressed highest in the brain,followed by the eye,liver,heart,fat tissue,kidney and gill. For adultM. amblycephala,theSREBP1 gene expressed highest in the brain,followed by the gonad,muscle,liver,eye,while expressed relatively low in the kidney,spleen and heart. The tissue expression pattern of the SREBP1 gene was different between the juvenile and adult M. amblycephala.

Abstract:To evaluate muscle development and fat deposition of blunt snout bream from Liangzi Lake and Yuni Lake,we detected histomorphological characteristics of the fat and liver,and the variation of fat accumulation by H&E staining,assessed the expression of the genes related to growth rate and lipid metabolism by qPCR.We found that fat accumulation in the mesenteric adipose tissue of blunt snout breams from Yuni Lake was about 1.5fold of that from Liangzi Lake.H&E staining results showed that the diameter of lipid droplet was shorter in the adipose and liver tissue of blunt snout breams from Liangzi Lake than that from Yuni Lake.The expression of LEP in the muscle and adipose tissue of blunt snout breams from Yuni Lake was prominently lower than that from Liangzi Lake (P<0.05),but IGF1 mRNA level was dramatically higher in the muscle of blunt snout breams from Liangzi Lake (P<0.05).The expression of LPL and NPY showed no significant difference in the muscle and adipose tissues of blunt snout breams from Liangzi and Yuni Lake (P＞0.05).From above,we proposed that the capacity of fat accumulation and key genes’ (LEP and IGF1) expression were different in blunt snout breams from two lakes,which may result to the different growth rates of blunt snout breams between two lakes.

Abstract:In order to obtain SNP markers associated with hypoxia tolerance trait for applying in molecular markerassisted breeding of Megalobrama amblycephala,in the present study,based on the constructed transcriptome data in hypoxia tolerant and sensitive M. amblycephala,52 623 potential SNPs including transition type of 30 192 (57.4%) and transversion type of 16 802 (31.9%) were found. Synonymous SNPs accounted for 99.7% of total SNPs located in the coding region,while only 32～35(0.3%) nonsynonymous SNPs were found. Five nonsynonymous and polymorphic SNPs were obtained using PCR-RFLP method. Analysis showed that two SNPs,Plin2 A1157G and Hif3α A2917G,were significantly associated with hypoxia tolerance trait in the parental populations. But further analysis revealed that no significant association existed between the two SNPs and hypoxia tolerance trait in the offspring populations. It is indicated that molecular markers isolated in the parent populations may be inappropriate in their offspring,therefore,the effectiveness of markers in offspring needs to be considered in molecular marker assisted breeding.

Abstract:To explore the possible function of the complement factor B and C2 (Bf/C2) gene in blunt snout bream (Megalobrama amblycephala), the cDNA sequences were cloned with RT-PCR and the expression of the Bf/C2mRNA in healthy fish tissues and after Aeromonas hydrophila infection during early development was investigated with quantitative realtime PCR.The results showed that the sequence length of Bf/C2A cDNA was 2 520 bp containing a 42 bp 5′ UTR,a 2 298 bp ORF and a 180 bp 3′ UTR,encoding 765 amino acid.The Bf/C2B cDNA had an 2 131 bp ORF,encoding 710 amino acid.The mRNA of both Bf/C2A and Bf/C2B were highly expressed at 1 dpf,followed by the intestinal formation stage during early development.The highest expression of both Bf/C2A and Bf/C2B in healthy adults appeared in the liver,followed by the kidney,head kidney and spleen.Theexpression of Bf/C2A and Bf/C2B exhibited a significant change after infection of A. hydrophila in the inmmunityrelated tissues including the liver and kidney,indicating that Bf/C2 plays important role in the immune response against bacterial infection.

Abstract:The genomic sequences of hepcidin in Megalobrama amblycephala were obtained from public database to study the association between the hepcidin gene polymorphism and disease resistance trait. Aeromonas hydrophila challenge experiment was used to distinguish susceptible and disease resistant M. amblycephala individuals.The single nucleotide polymorphisms (SNP) of the hepcidin gene in M. amblycephala were screened with PCR and DNA sequencing. Highresolution melting (HRM) were used for SNP genotyping. Two SNPs of the hepcidin gene were screened,one of which was located in the intron region and the other was located in the 3′untranslated region. The results of statistical analyses showed that the genotype and allele frequencies of SNP (A/G) located at 3 371 bp was extremely significant different (P<0.01) and SNP (A/G) located at 217 bp was significant different (P<0.05) between the susceptible and resistant group. It is indicated that the polymorphism of hepcidin gene was significantly associated with bacterial septicemia resistance.

Abstract:In order to reveal the change in photosynthetic characteristics and responsive mechanisms of different leaf positions in watersaving droughtresistant rice and highyield rice under watersaving irrigation,watersaving and droughtresistant rice Hanyou113 (HY113) and highyield rice Yangliangyou 6 (YLY6) were used to conduct a pot experiment in 2015.The effects of traditional flooding (F),saturated treatment (S) and drought treatment (D) on the leaf water potential,SPAD values,leaf gas exchange,instantaneous water use efficiency of function leaf at four important growth stages including jointing stage,booting stage,heading stage and milky stage and yield of rice varieties were studied.The results showed that the yield of YLY6 in S treatment was significantly increased compared with other treatments,contributed by higher effective panicles,grains per panicle and filled grains rate.Correlation analyses showed that there was a significant positive correlation between dry matter accumulation and yield heading stage and milky stage,and a significant positive correlation between net photosynthetic rate at the booting stage and heading stage and the dry matter quality at heading stage and milky stages.The dry matter accumulation of YLY6 at booting stage,heading stage and milky stage under saturated treatment was significantly higher than that of HY113.The instantaneous water use efficiency of YLY6 at heading stage was significantly higher than that of HY113.Under S treatment,YLY6 maintained high net photosynthetic rate with high SPAD value,stomatal conductance,and transpiration rate at the postfertility stage (from milky stage to mature stage) to maintain a high photosynthetic productive potential.Therefore,YLY6 maintains high photosynthetic capacity in different leaf positions at the late growth stage (milky period) under S treatment,which is the basis for watersaving and highyield rice.

Abstract:The effects of pumpkin rootstock (‘Qingyan No.1’) grafting on growth,chlorophyll fluorescence,photosynthetic gas exchange,expression of Calvin cycle and stresstolerance genes of watermelon (‘97103’) seedlings under chilling stress (10℃/5℃) were studied to clarify the mechanism of photosynthetic adaptation involved in chilling tolerance in pumpkin rootstockgrafted watermelon seedlings.The results showed that the chlorophyll fluorescence parameter such as maximum quantum yield of PSⅡ (Fv/Fm) in selfgrafted watermelon decreased significantly under chilling stress in comparison with control.After 5 days of chilling stress,the effective quantum yield of PSⅡ (ΦPSII) in selfgrafted watermelon seedlings and pumpkin rootstockgrafted seedlings decreased by 92.9% and 59.2% compared with the control.Pumpkin rootstock grafting effectively delayed the damage of PSⅡ induced by chilling and promoted the expression of key genes including Rubisco large subunit (rbcL),glycerate 3phosphate kinase (PGK),phosphoribulose kinase (PRK), and Fructose1,6bisphosphatase (FBPase) involved in Calvin cycle.A higher CO2 assimilation rate (Asat) in pumpkin rootstockgrafted seedlings under chilling stress was observed.which was attributed to the enhanced expression of 9 stresstolerance genes including GPX, tAPX, GR, and ERD10.After 5 days of chilling treatment,the chilling injury index of selfgrafted watermelon seedlings and pumpkin rootstockgrafted seedlings were 61.1% and 22.2%,respectively.The whole plant dry weight was decreased by 61.0% and 33.2%,respectively compared with the control.It is indicated that pumpkin rootstock grafting reduces the water loss by regulating transpiration,upregulates the expression of key genes involved in Calvin cycle and stresstolerance,thus alleviates photoinhibition of PSⅡand reduces the damage of membrane lipid peroxidation to enhance the adaptability of watermelon under chilling stress.

Abstract:Thirty three Brassica materials including four Brassica junceaCoss.var. megarrhiza Tsen et Lee,one B. juncea var. crassicaulis Chen et Yang,three B. juncea var. tumidaTsen et Lee,six Brassica juncea var. multicepsTsen et Lee, eleven Brassica juncea var. rugosa Bailey,two Brassica junceavar.capitata Hort,one Brassica juncea var. longepetiolata Yang et Chen,one Brassica junceaCoss.var. gracilis Tsen et Lee,four Brassica juncea var.latipa Li were inoculated with TuMV isolate.Through disease investigation and convulsion ELISA ,nine mustard materials were found to be TuMVresistant,laying the foundation for disease resistance breeding.The condition indexes of both of the numbers 17Y7V and 17Y36V in disease investigation were 0, showing immunity.Its DP/DN ratio of the ELISA test were less than 2.1, which were 1.38 and 1.97,showing negative performance.The condition index of the numbers 17Y13V, 17Y20V, 17Y21V, 17Y31V, 17Y33V, 17Y34V, and 17Y35V in disease investigation were 2.86, 5.33, 2.00, 8.00, 5.93, 5.71, 2.00, showing high resistance. Its DP/DN ratio of the ELISA test were less than 2.1, which were 1.12, 0.89, 1.60, 0.93, 0.78, 0.81, and 1.33, showing negative performance.

Abstract:Aluminum toxicity is one of the main factors limiting the growth of crops in southern acid soils.In this experiment,the differences and potential mechanisms of aluminum toxicity alleviation effects of different modifiers on acidaluminum enriched soils were studied by adding exogenous aluminum and planting crops in red soil.Pot experiment with acid red soil,adding exogenous aluminum,and applying anendment humic acid(HA),biochar(C) and quicklime(CaO),planting pakchoi,setting 8 treatments including CK,HA (0.3% HA),C (2% C),CaO(0.3% CaO),Al(1 mmol/kg Al3+),Al+HA (1 mmol/kg Al3++0.3% HA),Al+ C (1 mmol/kg Al3++2% C),Al+CaO (1 mmol/kg Al3++0.3% CaO) was conducted to analyze the effects of humic acid,biochar and quicklime on different forms of active aluminum and crop growth in aluminum rich red soil.The results showed that aluminum toxicity significantly decreased the emergence rate and fresh weight of pakchoi,and increase the aluminum content in pakchoi.The growth of pakchoi improved and the Al content reduced under amendment addition.Compared with Al treatment,the fresh weight of Al+HA,Al+C,Al+CaO treated pakchoi reduced by 86%,775% and 1 014%,respectively.The pH of Al treatment decreased by 0.17 compared with CK treatment,the content of active aluminum significant increased,the amount of active aluminum decreased after added the amendment.The exchangeable Al3+ of Al treatment increased by 11.17% compared with CK treatment,but the exchangeable Al3+ of Al+HA,Al+C and Al+CaO treatment decresed by 11.81%,59.63% and 87.82% compared with Al treatment.The content of dimer hydroxyl aluminum ion Al(OH)2+,Al(OH)+2 increased.The exogenous aluminum significantly reduced soil pH.The content of exchangeable Al3+ increased,inhibiting the growth of pakchoi.Humic acid,biochar and calcareousness reduced the exchangeable Al3+ content.Biochar and calcareousness effectively reduced the pH of red soil and improved the growth of pakchoi.It is indicated that anendment affects crop growth by combining soil pH,nutrient content and exchangeable Al3+ content.

Abstract:The agnH gene was cloned and expressed in the nicotinedegrading Agrobacterium tumefaciens SCUEC1 strain,and the conditions of expressing and purifying AgnH protein were optimized.The full length agnH gene (585 bp) was amplified by PCR,and the recombinant plasmid pET28a(+)agnH was constructed and transformed into E. coli BL21(DE3) strain for heterologous expression.The effect of different IPTG concentration,induction temperature and induction time on the expression of recombinant plasmid pET28a(+)agnHwas studied.The recombinant protein was eluted with different concentrations of imidazole eluant,and the expressed protein was detected with SDS-PAGE.The results showed that the agnH gene was cloned and the recombinant plasmid pET28a(+)agnH was constructed.The expression of AgnH protein was higher under the conditions of IPTG concentration of 0.4 mmol/L,induction temperature of 25℃ and induction time of 25 h.A higher concentration of AgnH protein was obtained by elution with a 250 mmol/L imidazole eluant.

Abstract:The Escherichia coli K88, K99, 987P strains and C type Clostridium perfringens were used to prepare piglets colibacillosis. Pregnant sows were immunized with different doses of vaccine, and then their piglets were challenged with the corresponding pathogenic strains and doses. The results showed that the protection rate of the piglets was above 80% when the vaccine doses was 1.6 mL or more. The protection rate of piglets against bacteria was not less than 80% when piglets aged 1, 4,7 and 10 d were challenged, which indicated that the immunization period was within 24 h after colostrum feeding and the duration of passive immunization was more than 7 d. Three batches of vaccines were kept at 28℃ and tested at different times during preservation. The results showed that the vaccine was safe for pregnant sows after 27 months of preservation and the protective rate of piglets produced by the vaccine immunized sows was no less than 80%. Therefore, the minimum immune dose of the vaccine was 1.6 mL, the passive immunization duration of piglets was more than 7 d, and the vaccine storage period was 24 months at 28℃.

Abstract:The gel strength,water holding capacity,whiteness,interactions and microstructure of silver carp surimi gels were measured after secondary heating treatment.The effects of different secondary heating treatment conditions ( 70~100℃ for 30 min and 60 min ) on the quality of silver carp surimi gels were investigated.The results showed that the gel strength,water holding capacity and the contents of disulfide bonds and hydrophobic interactions of the surimi gels increased firstly and then decreased with the increase of the secondary heat treatment temperature and time compared with the control group.Especially for 60 minutes heating at 100℃,its gel properties were completely destroyed.When the temperature of the secondary heat treatment was higher than 80℃,the threedimensional network structure of the surimi gels were destroyed to a certain extent,and the degree of destruction increased with the increase of the secondary heat treatment temperature and time.This result was consistent with the trend of the above indicators.The results of SDSPAGE patterns showed that the myosin heavy chain (MHC) bands decreased gradually with the increase of the secondary heat treatment temperature and time.When the secondary heat treatment temperature reached 100℃,the MHC bands almost disappeared.It will provide a theoretical basis for further understanding the thermal stability of surimibased products.

Abstract:Aiming at the problem of noise interference in egg crack detection process,this paper collects the audio vibration signals of the eggs on the transportation. Drawing recurrence plot(RP) of audio vibration signals which are unprocessed and using recurrence quantification analysis(RQA) to extract the quantitative feature parameters of recurrence plot. These quantitative feature parameters are recurrence ratio,determinism,laminarity,entropy and maximum diagonal length. Using these parameters to detect whether eggs are cracked. Results showed that the accuracy of detection and classification of egg with cracks is verywell via a support vector machine (SVM)，back propagation neural network(BPNN) models. 300 eggs were detected in this study. The results showed that the SVM model was better, in the SVM model,the recognition rate of intact eggs and crack eggs was 93.98% and 95.52%.

Abstract:Infield rapeseed seedling images under natural illumination were studied with six colorvegetationindex segmentation approaches including excess red vegetative index (ExR),excess green vegetation index (ExG),excess green minus excess red (ExGR),normalized difference vegetation index (NDI),color index of vegetation extraction (CIVE) and combination of vegetation indices (COM).The thresholdingbased algorithms were used to extract infield rapeseed plant with shadow image.The segmentation of common RGB vegetation indices were objectively estimated with quantitative evaluation criteria.The results showed that the COM index is superior to the other 5 vegetation indices in the qualitative analysis,which can reduce the segmentation effect caused by the shadow and retain the complete blade profile in the local blade segmentation tests.In the quantitative analyses,the COM index provides the best segmentation accuracy,sensitivity and specificity of 94.1%,97.2% and 90.9%,respectively,with the corresponding standard deviations of 1.1,1.3 and 0.06.

Abstract:In order to research the influence of the rack tooth forms and working conditions on the mechanical property of selfpropelled monorail mountain orchard transporter，the dynamic models of the transmission system including the driving wheel and racks were established based on the dynamic theory and the Hertz impactcontact theory.Based on ADAMS/View simulation software,the relation of the tooth forms and the torque of the driving wheel which was generated when the driving wheel was engaged with the racks was analyzed and simulated.The orthogonal experiment was carried out using tooth forms,rail gradients and angular velocity as experiment factors and the average torque as assessment index.Range analysis and variance analysis was used to determine the importance index for the torque.The results showed that the order of the primary and secondary impact on the mechanical performance of the transporter is rack tooth profile>track grade>drive wheel speed.Under the same conditions,the sprocket rack has the best performance.Under the condition of the angular velocity of the drive wheel was +88.08 rad/s,the rail gradients were +0°,+6°,+12°,the average torque was decreased by 33.82%，33.45%，18.36%,respectively when the driving wheel was engaged with chain wheel tooth form rack than arc tooth form rack.The correctness of the model simulation analysis is verified,which will provide a reference for the structural optimization design of selfpropelled monorail mountain orchard transporter track.

Abstract:A rigidflexible virtual prototype model of the manmachinepavement system of the harvester was established by combining finite element analysis software with the software of SolidWorks and RecurDyn.Weighted root mean square values of acceleration was used to evaluate the vibration comfort of the harvester.On the basis of the virtual prototype model of the system,the vibration comfort of the harvester was simulated when it harvests the rice,wheat and rapeseed.The results showed that the driving speed,the road surface hardness and the vertical cutting knife influence the vibration comfort of the harvester greatly.With the increase of driving speed of the harvester,the weighted acceleration RMS of the mass center of human body in different axis and the wholebody vibration acceleration RMS increased rapidly,enhancing the subjective discomfort feeling of the driver.In addition,the vertical cutting knife enhanced the discomfort feeling of the human body.The subjective discomfort feeling of the drive enhanced with adding the vertical cutting knife.

Abstract:In order to improve the automation and intelligence level of sows breeding equipment in China,an intelligent and precise feeding equipment for groupbreeding sows is designed,which consists of all mechanical feeding channels,precision feeders and intelligent controllers. The device adopts a small number of feeding ways and a programmable controller as the core controller to collect and store the feeding information of sows. The performance measurement analysis showed that when the single discharge amount was 100 g,the average relative error of the daily discharge amount of the device was 2.59%,the mean coefficient of variation was 2.25%,and the maximum identification radius was 16 cm. The results show that the device has the functions of identifying feeding sows accurately,isolating feeding and precise feeding. The accuracy and stability of feeding and recognition range of the device meet the design requirements.