Abstract:HL- sp1 , a 6.7 kb HL-CMS-related fragment, was cloned from the mitochondrial genome of HL-CMS sterile line YTA. One of the two ORFs predicted encodes 290 amino acids and is tentatively named orf290 . One overexpression vector 35S∶∶Rf1b5′∶∶orf290 with a mitochondrial signal peptide was constructed and transformed by Agrobacterium into YTB, a HL-CMS maintain line. The copy numbers of the exogenous gene in the 35S∶∶Rf1b5′∶∶orf290 transgenic T0 plants were estimated according the ratio of hygromycin B phosphotransferase ( hpt ) gene and the single copy sucrose phosphate synthase ( SPS ) gene in rice with qPCR based on DNA. The pollen fertility and the seed set rate of the T0 positive transgenic plants were investigated. The results showed that two transgenic plants with single copy of 35S∶∶Rf1b5′ ′∶∶ orf290 were obtained and the expression level of orf290 in the T0 positive transgenic plants were detected with qPCR. The average pollen fertility was 30.8% and the average seed set rate was 41.6% in the single copy transgenic lines. It is indicated that mitochondrial gene orf290 may be related to cytoplasmic male sterility in rice.

Abstract:The relationships between fruit granulation and low temperature in winter of different years in Zigui County of Three Gorges Reservoir Area were studied. The changes of fruit quality and fruit granulation in the late maturing navel orange fruit ripening period (from December to June) at different altitudes (520 m,380 m,220 m) in Zigui County of the Three Gorges Reservoir were investigated. The effect of the navel orange fruits bagging in the winter to prevent fruit granulation was studied. Results showed that fruit granulation of the late maturing navel orange in the Rservoir Area was mainly caused by the low temperature in winter. Fruit quality of high altitude (520 m) was significantly lower than that of middle and low elevations (380 m,220 m). At high altitude (520 m) ,the soluble solids content and solid acid ratio,water content,juice rate were low,and the fruit color was poor. Fruit water content and fruit granulation decreased with the harvest stage. The ratio of fruit granulation was high at the stage of flowering (late April) and shoot growth (early May),reaching 50.0% and 38.8%,respectively. The index of fruit granulation increased. The degree of fruit granulation aggravated gradually. A few fruit slightly granulated in middle June at low elevations. Fruit granulation didn’t occur in other time at low elevations.Fruits bagging helps reduce or prevent fruit granulation. The rate of fruit granulation decreased to 6.0%,while the content of fruit edible rate,fruit juice rate and soluble solids increased significantly,and fruit coloring and internal quality was improved significantly as well. 

Abstract:A loop-mediated isothermal amplification (LAMP) assay was developed for rapid and accurate detection of Pratylenchus brachyurus , one of the important plant pathogenic nematodes. Based on the mtDNA COI sequence, LAMP primers for specific detection of P. brachyurus were designed. The optimal conditions of the LAMP assay was the reaction time was 60 min at 64℃ . The LAMP products of P. brachyurus were confirmed using conventional polymerase chain reaction (PCR), enzyme digestion analysis with Bam HⅠ and visual inspection by adding SYBR Green I to the products. P. brachyurus from nine Pratylenchus species and other ten plant-parasitic nematode species were successfully detected by the LAMP assay. The LAMP assay was also sensitive, which can detect DNA from 0.005 individual P. brachyurus , 10 times more sensitive than conventional PCR. The LAMP system constructed in this study can be used to detect P. brachyurus rapidly. 

Abstract:Microalgae is the third generation of potential bio-energy resources,and the presence of grease in microalgae is mainly triacylglycerol.Diacylglycerol acyltransferase ( DGAT ) is a key rate-limiting enzyme in the control of TAG synthesis. DGAT is divided into DGAT1 , DGAT2 , DGAT3 and WS/DGAT .The DGAT of microalgae and higher plants was obtained from the database.The diversity and evolution of the four DGAT types and gene and protein structures was analyzed with phylogenetic methods.In order to further analyze the evolution of DGAT in microalgae,the positive selection sites were selected to estimate the selection pressure in the evolutionary process.Result showed that the genetic structure and protein structure of different DGAT types in microalgae were different.In the same DGAT type,the gene structure and protein structure of microalgae was different from that of other higher plants.It was found that microalgae did have different evolutionary processes with other higher plants.There was a positive selection sites of DGAT2 and WS / DGAT .Thepositive selection sites of DGAT1 and DGAT3 were not found.It will help us to decipher its functional role in plants and promote the research of renewable energy.

Abstract:Microalgae is the third generation of potential bio-energy resources,and the presence of grease in microalgae is mainly triacylglycerol.Diacylglycerol acyltransferase ( DGAT ) is a key rate-limiting enzyme in the control of TAG synthesis. DGAT is divided into DGAT1 , DGAT2 , DGAT3 and WS/DGAT .The DGAT of microalgae and higher plants was obtained from the database.The diversity and evolution of the four DGAT types and gene and protein structures was analyzed with phylogenetic methods.In order to further analyze the evolution of DGAT in microalgae,the positive selection sites were selected to estimate the selection pressure in the evolutionary process.Result showed that the genetic structure and protein structure of different DGAT types in microalgae were different.In the same DGAT type,the gene structure and protein structure of microalgae was different from that of other higher plants.It was found that microalgae did have different evolutionary processes with other higher plants.There was a positive selection sites of DGAT2 and WS / DGAT .Thepositive selection sites of DGAT1 and DGAT3 were not found.It will help us to decipher its functional role in plants and promote the research of renewable energy.

Abstract:Flow cytometry and spectrophotometry were used to measure the cell numbers and absorbances of three common cultures including Escherichia coli S17-1, Synechocystis sp. PCC 6803 and Bacillius subtilis 168 at different concentrations in the laboratory. Then the absorbance values and cell numbers of each culture were compared. The function relationship between absorbance and cell numbers was established. The results showed that the absorbance value and cell numbers of the three common cultures in the laboratory were significantly positively correlated with the concentration at the certain concentration. It was verified that the absorbance value of the culture can reflect the number of cells during the culture process using the fitting function. The relationship between the absorbance and cell number of three strains including Escherichia coli S17-1, Synechocystis sp. PCC 6803 and Bacillius subtilis 168 was established. It will provide the foundation for developing biological experiments to determine the cell number and downstream molecules in the culture using a spectrophotometer. 

Abstract:Microbiome sequencing technology was used to study the effects of applying 4 kinds of straw compositing agents including T1( Trichiderma asperellum/Bacillus subtilis ),T2( Aspergillus niger/Bacillus subtilis ),T3( Rhizopus nigricans / Bacillus subtilis ) and T4( Trichiderma asperellum / Bacillus subtilis ) on soil microbial ecology.The results showed that the population of fungi was more affected by the treatments,in which the number of family and genus was obviously increased.The number of family and genus in the T2 treatment was the biggest (133.3%) compared to positive control (Tp).The number of family or genus in the T4 treatment increased with a small range from 15.6%-45.8%.The population of bacteria was not largely changed.Results of comparing Chao1 index showed that the fungi index dramatically increased with a range from 74.5% to 191.8%,whereas bacteria index was changed at a small scale (-4.0%-15.1%).Results of statistics analyses showed that 24 to 55 genera of fungi in different treatments were significantly up-graded ( P <0.05 and fold changes≥2),of which more than 60% were commonly significantly up-graded in contrast to both Tp and Tn,in which only 2-11 genera of bacteria were significantly up-graded.4-11 genera of fungi were significantly down-graded,of which 14.3%-25.0% were commonly significantly up-graded in contrast to both Tp and Tn,in which 4-7 genera of bacteria were significantly down-graded related to Tp,but none was down-graded compared to Tn.The similarities of microorganism Beta diversity in different treatments were analyzed by UPGMA cluster.The results of soil fungal Beta diversity showed that T1,T2,T3 and positive control were in the same cluster,while T4 and negative control in another cluster. The results of bacterial Beta diversity showed that T2,T3,T4 and positive control were in the same cluster,while T1 and negative control were in the same cluster.

Abstract:Knowledge of the interaction characteristics between soil humic substance and carbonate is important for better understanding of the coupling relationship between soil organic carbon and inorganic carbon.Soil humic substance from Jiugong Mountain in Hubei and synthetic calcite were selected for studying the adsorption of humic acid (JGHA)/fulvic acid (JGFA) on calcite and its dissolution process.The interaction characteristics and influence factors were illuminated by comprehensive analysis of JGHA/JGFA before and after adsorption and JGHA/JGFA-calcite complex.The results showed that a preferential adsorption of humic substance components with higher molecular weight and complex structure on calcite surface has occurred.Sorption isotherms of JGHA/JGFA on calcite followed Langmuir type behavior with a correlation coefficient R2＞0.95.The adsorbed JGHA amount was higher than that of JGFA.The adsorption density of JGHA was far lower than that of JGFA.The adsorbed amount of JGHA/JGFA on calcite decreased with increasing pH which signified a strongly pH-dependent.The pH-dependent of JGHA is stronger than that of JGFA and increased with increasing humic substance concentration.Dissolution of calcite was inhibited by the absorbed hydrophobic molecules of JGHA on the calcite surface,while JGFA with lower molecular weight and higher amount of acidic functional groups promoted the dissolution of calcite.The effect of JGHA/JGFA on calcite dissolution was enhanced with increasing humic substance concentration.Adsorption and sequestration of organic carbon and its stability,dissolution and precipitation of inorganic carbon in arid and semi-arid soil rich in cacium carbonate are depended on the molecular properties and functional group composition of soil organic matter.

Abstract:In order to explore the genetic background of short gestation period and high litter size of Qingping pigs, correlation analysis between ESR , FSHβ and PRLR genes and the reproductive performance in F1 population and F2 black pigs of Qingping pigs × Landrace hybrid was carried out. The results showed that: (1) The gene frequency of A and B allele of FSHβ was similar in both F1 and F2 populations, while the predominant allele of ESR in both populations were B allele. The predominant allele of PRLR in F1 populations was C allele, and gene frequency of C and T allele in F2 population was similar. (2) Different genotypes of FSHβ had significant effects on the number of live piglets in F1 population, and on the total number of litters, birth weight, weaned piglet, and weight at 3 weeks of age in F2 population ( P <0.05). (3) Effects of different ESR genotypes on gestation period in F2 were in the order of AB

Abstract:To identify the methylation pattern of the porcine MKRN3 promoter CpG island in the embryo heart,embryos of days 65 and 100 of gestation from the reciprocal crosses of Yorkshire-Meishan pig were collected.Firstly,CpG island in the promoter region of MKRN3 was predicted by utilizing bio-informatics method,and primers was designed based on this predicted CpG island.Bisulfite sequencing (BSP) was utilized to detect methylation level in the embryo heart from Yorkshire-Meishan reciprocal crosses.Results showed that,in the heart from embryo of days 65,the CpG island of porcine MKRN3 showed high level of methylation,with 75.6% from Yorkshire×Meishan and 76.4% from Meishan×Yorkshire.In the heart from embryo of 100 days,the CpG island of target gene showed high level of methylation (80%) from Yorkshire×Meishan,while low level of methylation (35.6%) from Meishan×Yorkshire crosses.Results above indicated that the methylation pattern of the porcine MKRN3 promoter CpG island would change according to the change of reciprocal crosses and developmental stages.For embryo of 65 days,reciprocal crosses of Yorkshire-Meishan represented high methylationlevel,but for embryos of 100 days,high methylation from Yorkshire×Meishan but low methylation from Meishan×Yorkshire were observed,indicating that de-methylation might take place in the paternal or maternal allele,during the late stage of embryonic development.

Abstract:In order to understand the clinical antimicrobial resistance of Staphylococci and their multidrug resistance gene cfr from animals in Xinjiang,and to provide guidance for clinical medication and further study of the cfr gene transmission mechanism,a total of 1 781 animal feces and nasal samples were collected from some areas of Xinjiang Uygur Autonomous Region.A total of 1 667 strains of Staphylococci were isolated from the samples,and then drug sensitivity tests of 11 kinds of clinical antibiotics were carried out.The PCR method was used to detect the amide-alcohol resistance genes ( cfr , fexA ,and fexB ),the mobile elements IS 21-558 and transposase gene Tn 558 ,which were related to the transfer of cfr .The results showed that the order of severity of drug resistance of Staphylococci to clinical commonly used antimicrobial agents was sheep source＞pet source＞pig source＞poultry source＞cattle source. The penicillin resistance was the most serious among the tested antibiotics,and the drug resistance rate was between 54.7% and 93.6%. Except for Staphylococci isolated from cattle (36.3%),the resistance rate of Staphylococci isolated from other animals to oxacillin was 51.1%-98.2%. Staphylococcus from animals showed good sensitivity to amikacin,gentamicin and clindamycin. Among the 11 clinical antibiotics,the multidrug resistance rate of Staphylococcus was concentrated in 4-5 from the sheep source,was concentrated in 3-5 from the poultry and pet source,was mainly 4 and 7 from the pig source, and was 1 from the bovine source. Staphylococcus isolated from different animal sources had a wide range of antibiotics resistance spectrum. The detection results of amide-alcohol resistance gene showed that,only 4 strains of Staphylococcus isolated from pigs carried cfr gene. Except for bovine source Staphylococcus , fexA gene was detected in other animal sources with the order of detection rate of pet source＞pig source＞poultry source＞sheep source,but the fexB gene was not detected. The 16S rRNA sequence amplification results showed that the 4 strains carrying the cfr gene were all Staphylococcus simulans . In addition,mobile element IS 21-558 (including istAS and istBS ) and Tn 558 transposase gene ( tnpA , tnpB , tnpC ) were detected in 4 positive bacteria. From above,the resistance of Staphylococcus from animal origin was serious in Xinjiang,the spectrum of resistance was diverse,and the main gene of amide-alcohol resistance was the fexA gene. In addition,multidrug resistance gene cfr and its mobile components were detected. It is suggested that the surveillance of drug resistance should be strengthened to prevent the outbreak of drug resistance bacteria and the probable emergence of super resistant bacteria.

Abstract:In this study,the effects of β-pinene on spore germination,tube elongation and mycelial growth of Penicillium italicum were assessed,and the effects of β-pinene on cell wall chitin distribution,active oxygen metabolism,acid excretion capacity and ergosterol contents of mycelia were analyzed.The effects of sorbitol,cholesterol and cysteine as protective agents on spore germination of P. italicum incubated with β-pinene were further evaluated.The results showed that β-pinene had strong antifungal activity against P. italicum in a dose dependent manner,as exhibited by spore germination and mycelium growth inhibition at the concentration ranging from 0.087 to 0.696 mg/mL.β-Pinene incubation also altered the distribution of chitin in cell wall,promoted the accumulation of reactive oxygen species (ROS),and significantly inhibited glucose-induced reduction in external pH of the mycelia.Addition of cholesterol as alternative of ergosterol,sorbitol as osmotic regulator,or cysteine as active oxygen scavenger could effectively promote the spore germination and tube elongation of the pathogen incubated with β-pinene at 0.348 mg/mL.These results revealed that the antifungal activity of β-pinene against P. italicum was related to the damage of cell wall structure,promotion of ROS accumulation and the inhibition of ergosterol increase in membrane.

Abstract:An isocarbophos molecularly imprinted solid-phase microextraction (MISPME) coating was prepared by sol-gel technology with polyethylene glycol as a monomer and isocarbophos as a template,and a headspace solid phase microextraction method coupled with gas chromatography (HS-SPME/GC) for the determination of isocarbophos in fruits and vegetables was established. Results showed that the imprinted coating had very good selectivity to isocarbophos,and its extraction capability was better than the non-imprinted fiber and commercial fibers. At the optimum conditions,the proposed method was linear in the range of 10-10 000 μg/kg with a coefficient of 0.999 6. The limit of detection was 5.6 μg/kg. The relative standard deviation of 5 replicates was 2.29%. The method was used for the determination of isocarbophos in 5 kinds of real samples including cucumber,tomato,apple,pear and cabbage. The results indicated that no residue of isocarbophos was detected in these samples. Satisfactory recoveries of 89.4%-106.6% were obtained in these matrices,indicating the accuracy of the method. 

Abstract:Transglutaminase (TGase) from silver carp was purified with ammonium sulfate precipitation,gel filtration on Sephacryl S-300 high resolution and DEAE-Sepharose fast flow ion-exchange chromatography. Some kinetic properties of transglutaminase purified were studied. The results showed that the molecular mass of the purified enzyme was 100 ku. The enzyme exhibited final purification fold of 34.2,specific activity of 126.7 U/mg. Results of further assaying enzymatic characterization showed that the purified enzyme had maximal activity at 30℃ and pH 7.5. The activity of transglutaminase was slightly activated by Ca2+ and dithiothreitol (DTT) and strongly inhibited by ethylenediaminetetraacetic acid (EDTA). The metal ions Mg2+,Cu2+,Zn2+,Ba2+,Sn2+,Fe3+ strongly inhibited transglutaminase activity as well. The transglutaminase can catalyze covalent cross-linking of mysion heavy chain (MHC) from silver carp and grass carp. The effects of the transglutaminase on mysion from silver carp were more noticeable than that from grass carp.

Abstract:The common nutrient compositions,minerals and amino acids of Agaricus taeniatus were detected and compared with other mushroom species including the wild Morehella esculenta , Boletus edulis and cultivars Lentinus edodes , Agaricus bisporus . The international general nutritional value assessment method was applied for the overall nutritional evaluation of its protein.The results showed that the content of crude protein in Agaricus taeniatus was 41.6%. The mineral contents were rich in potassium and magnesium,higher than tested species. Agaricus bisporus contained 16 kinds of amino acids (tryptophan was not tested),and the contents of glutamic acid,aspartic acid,alanine accounted for 40% of the total amino acid content. The amino acid score (AAS),chemical score (CS),essential amino acid index (EAAI),biological value (BV),nutritional index (NI) and score of ratio coefficient of amino acid (SRCAA) of protein were 61.47,39.34,65.55,59.75,27.27 and 70.86,respectively. It is indicated that Agaricus taeniatus is a kind of delicious mushroom with high nutritional value.

Abstract:A pneumatic insertion picking-up device is developed to solve the problem that the structure and seedling process of mechanical picking-up device for paper seedling transplanter are complicated. The mechanical model of seedling process was constructed. Key parameters of each seedling stage were optimized and the optimal parameter combination of the plant was obtained. The 60 mm×30 mm×60 mm(above length×below length×height) pour four square table shaped paper pot was used as seedling pot. A kind of double needle insertion type terminal actuator was designed. According to the requirement of the spacing of the needles,the model of the actuating cylinder is MHS4-20D. It was confirmed that the pneumatic picking-up process can be divided into stage of seedling,feeding and seedling removing. Results of mechanical analysis and parameter optimization test of each stage showed that when the friction between seedling needles and seedling pot matrix is not less than 3.67 N with dropping height of seedling pot less than 380 mm,the seedling picking-up device can effectively carry out the function of picking. The result of bench test showed that optimal parameters include the seedling needles are made of stainless steel with a diameter of 4 mm,inserting depth of 20 mm and dropping height of 340 mm,the success rate of seedling is 92%.The results of field testing showed that the effect of the pneumatic insertion picking-up device meets the technical requirements of the pot seedling transplanter. It will provide a reference for designing the pneumatic picking-up device and optimizing key parameters of the pot seedling transplanter.

Abstract:A machine vision image acquisition system was constructed to obtain 186 eggs hatching the 4 d image under LED light source to identify the early embryonic sex of chicken eggs. The preprocesses including component extraction,debackgrounding and binarization of egg image were carried out,followed by the using adaptive histogram equalization,top-hat and bottom-hat to enhance the image. Threshold segmentation of iterative add “and” operation was used to highlight the blood line texture. The 11-dimensional feature parameters of the image were extracted with the method of difference box,gray level co-occurrence matrix,gray histogram and geometric. When a BP (back propagation neural network,BPNN) model of chicken egg embryo sex identification was built,genetic algorithm (genetic algorithm,GA) was used to optimize BP neural network initial weights and thresholds initial. The results showed that the comprehensive accuracy of the training set of the GA-BP model was 99.73%,and the comprehensive accuracy of prediction set was 82.80%.