To prevent the vesicular stomatitis virus ( VSV) disease , using a panel of recombinant VSV (E１７６４A , S１８２７A , Y１６５０A , and F１６９１A) that were defective in mRNA cap methylation as vaccine to inoculate five‐week‐old BALB/c female mice through oral route with １ × １０６ PFU , the pathogenicity and immunogenicity of these viruses were determined . The result showed that recombinant S １８２７A , Y１６５０A , F１６９１A were attenuated in mice and E１７６４A was still pathogenic to mice . The immunized mice were challenged with wild type VSV . Mice immunized with S１８２７A and Y１６５０A trigged a high level of neutralizing antibody and were protected from virulent challenge . Taken together , the results above demonstrated that methyltransferase ( MTase)‐defective VSV ( S１８２７A and Y１６５０A) were not only attenuated in animals , but also exhibited excellent immunogenicity . Therefore , MTase‐defective viruses will be good live vaccine candidates against VSV .

Based on the nucleotide sequence of avian reticuloendotheliosis virus ( REV ) SNV strain ,Sichuan isolates REV-SC１ were used as a template , and a pair of primers was designed to amplify the １２１-１ ０２３ bp of gp90 gene fragment encoding high antigenic and hydrophilic domain of C‐terminal protein . The gp90 gene was inserted into pMD-１９T vector and sequenced , and the online software of NCBI blast was used to analyze the sequencing result . The gp90 gene was inserted to expression plasmid pET-３２a( ＋ ) , and the recombinant plasmid was transformed into Escherichia coli BL２１ (DE３ ) , then using IPTG to induce expression of protein , at last , an indirect enzyme‐linked immunosorbent assay (ELISA) method was established for detecting the antibody to REV . The results showed that :１) Compared with other REV strain nucleotide sequences , the maximum matching rate was ９９％ , and there was a mutation separately at the ２３１ and ８７０ sites . ２) SDS‐PAGE analysis showed that the expected protein was got with molecular mass of approximately ４５ ku , and the Western-blot result indicated good reactogenicity of the target protein . ３) An indirect ELISA was established by using the recombinant protein . The optimal antigen coating concentration was set as １ .５ mg/mL ^-１ , and the best anti-serum dilute degree was at １ ∶ ３２０ , and the cut-off value ( OD_{４５０ nm} ) was 0.126 . The detection results revealed that the method had good diagnostic sensitivity and specificity .

Protein structures , physical and chemical properties , and their functional domain structures encoded by duck peroxisome proliferators‐activated receptors‐α ( PPARα ) gene were analyzed using bioinformatics method . Sequencing and bioinformatics analysis showed that the duck PPA Rα cDNA sequence was １ ４３０ base pairs long with an open reading frame ( ORF) containing １ ４０７ nucleotides encoding a protein of ４６８ amino acids . The multiple sequence alignment showed that coding nucleotide sequences of the obtained duck PPA Rα gene had ９６％ , ９２％ , ７９％ , ７７％ , ７８％ homology with the corresponding chicken , T aeniopygia guttata , human , bovine and Equus caballus , respectively , and the deduced amino acid sequences had ９８％ , ９７％ , ８９％ , ８９％ , ８８％ identity with the corresponding species , respectively . Phylogenetic tree analysis of the amino acid sequences of duck PPAR α and other PPARα proteins in human and other animals showed that , among all the species mentioned above , the nearest relationship existed between duck and chicken . Further analysis on the structure and function of the duck PPARα protein showed that there were much more α helix than other secondary structure in the duck PPARα protein , besides , it had a short nuclear localization signal (KKNRNKC) , so that it could enter into nucleus . Moreover , a DNA binding domain which was composed of two C４‐type zinc fingers and a ligand binding domain was found in the PPARα protein .

To elucidate the character and functions of goose interferon‐alpha gene ( I FN‐α ) , the gene IFN‐α of Tianfu meat goose open reading frame ( ORF) was cloned into eukaryotic expression vector pcDNA３ .１ ( ＋ ) to generate the recombinant plasmid pcDNA３ .１‐goIFN‐α , which was then transfected into the COS‐７ cells by lipofectin transfection . Then , its expression and localization in COS‐７ cells were determined by indirect immunofluorescence , enzyme‐linked immunosorbent assay and Western‐blot assay , and the antiviral activity of supernatant from transfected COS‐７ cells was also determined . The results indicated that goIFN‐α protein was fist detected at hour １２ post‐transfection , which was rapid increase in the supernatant before ３６ h , thereafter it became slow . The recombinant plasmid could be
highly expressed in COS‐７ cells , and a protein of ３８ ku in transfected COS‐７ cells was also detected , which was larger than the predicted molecular mass of ２１ ku . The indirect immunofluorescence showed that the cells appeared yellow‐green fluorescence , the fluorescent intensity in which was the most highest nearby nucleus . The antiviral activities of supernatant against vesicular stomatitis virus ( VSV) and gosling plague virus (GPV) were obvious , as verified by method of cytopathic effect inhibition test .

With a F_２ population between a resistant parent and susceptible parent , SRAP analysis was carried to develop molecular markers linked to powdery mildew in cucumber . Sixty‐two pairs of primer combinations which had stable amplified polymorphic bands between resistant parent and susceptible parent were selected from ４３７ pairs of SRAP primer combinations . Further analyzing the F_２ segregating populations using these primers , one SRAP marker of Me1 /Em9 — 284 bp linked to the powdery mildew resistance was found , with 9.8 cM genetic distance . The marker can be used for cloning disease‐resistant gene and advancing efficiency of disease-resistant breeding on molecular marker assisted selection .

PCR primers were designed according to BaANS gene isolated from Brassica albograbra , and the gene , designated BcANS , was amplified from cotyledons DNA and cDNA of B . campestris var . purpurea , respectively . The gene sequence was submitted to NCBI with an accession number of GQ１２０５６２ . Genomic DNA was １ ６３７ bp in length with one ５６０ bp intron , and the complete coding sequence was １ ０７７ bp encoding ３５８ amino acids . RT-PCR results showed that BcA N S expressed in cotyledons and hypocotyls under light condition , and there was no expression under dark condition . BcA N S expressed under dark and light conditions in phosphorus-deficient medium . However , higher expression levels in cotyledons and hypocotyls were observed under light condition . Sequence alignment results revealed higher homology between BcANS and those of B . oleracea , B . j uncea and A rabidopsis thaliana . Low similarity was observed between BcANS and those of Gramineae plants , indicating their distant evolutionary relationships . Isolation , sequence analysis of anthocyanidin synthase gene in B .campestris var . purpurea laid the foundation for further study of gene function and anthocyanin biosynthesis mechanisms .

The amino acid composition and hydrophilism and hydrophobicity of β‐momorcharin were analyzed by DNAMAN software , and its basic features , secondary and tertiary structures , a coiled coil area and a PEST sequence were predicted by bioinformatics online websites ,and the surface potential and accessibility were analysed by bioinformatics software , then the structural quality analysis and detection evaluation on the modeling result were carried out by establishing a Ramachandran graph . The result showed that the β‐momorcharin comprised 249 amino acids and had isoelectric points of 9.19, and the hydrophilic area and the hydrophobic area were distributed in a staggered manner ; the β‐momorcharin belonged to stable protein as the extinction coefficient thereof was 280 nm , and the average hydrophilism was -0.138; the secondary structure comprised 42.57% of α‐helix , 20.48% of extended strand and 36.95% of irregular coil ; there were three sections of coiled coil areas , five sections of non‐PEST sequences , and no stable dipolymer or tripolymer coiled coil ; the space distribution of tertiary structure ,surface potential and accessibility of β‐momorcharin were modeled , and the Ramachandran graph detection showed that the structure of the model met the rules of stereochemistry .

Four varieties of Brassica napus , Xiangyou １５ , Huyou １５ , You ８８ and Huyou １７ , were crossed with a temporary maintainer ( TAM ) named HYZ1-TAM respectively and produced four F２ populations . In these populations , molecular markers associated with Bnms3 and Bnr f were used to select new temporary maintainer plants which were identified by test‐cross . Sixty‐two plants with TAM marker‐genotype were selected by marker‐assisted selection ( MAS) from three hundred and fifty‐five plants of four F_２ populations . Fifty‐seven plants ( with the accuracy of 91.94% ) were TAM genotype proved by test‐cross . Compared with traditional method , it is very efficient to select TAM by MAS . In the Bnms4 locus , all the four varieties have a same genotype which is ms4ms4 .

For the purpose of shade‐tolerant fern screening , six indicators were chosen , which were specific leaf area , leaf mass per area , chlorophyll contents , maximum photosynthetic rate , light compensation point , and light saturation point , then the phenotypic plasticity index ( PPI) and shadetolerant threshold (STT) for the six indicators were calculated based on collected literature data . The results showed that the PPI of ferns varied within a range of ０.７４-０.９６ . The STT values of each index were as follows : chlorophyll content of 1.9 mg.g^-1 , chlorophyll a/b ratio of ２.３ , leaf mass per area of 36 g.cm^-2 , maximum photosynthetic rate of 5.5 μmol.m ^-2 .s^-1 , light compensation point of 24 μmol.m ^-2 .s^-1 , and light saturation point of 460 μmol.m ^-2 .s^-1 . The STT values can be used as evaluation and selection of shade‐tolerant ferns in various ways .

To improve the use of lianas in urban river landscape , physico-chemical changes in five different lianas , such as Hedera nep alensis , Celastrus orbiculatus Thunb . , Rosahybrida ClimbingRoses , Jasminum mesnyi Hance , Euonymus f ortunei were analyzed during low temperature stress . Membrane permeability , superoxide dismutase ( SOD) activities and soluble protein contents in leaves of these five plants were measured at different temperatures . The results showed that membrane permeability , SOD activity and soluble protein contents all increased as the temperature lowered and the time prolonged , which indicated that the three factors could be used as the indexes to leaf cold resistance of the five plants . Among the five plants , E . f ortunei showed the highest cold resistance , while C .orbiculatus showed the lowest cold resistance than the others .

The growth habit , morphological and reproductive characteristics of Hydrocotyle vulgaris were systematically described to assess its invasion risks . The results showed that the invasion risks of H . vulgaris were displayed in four aspects . First , the adaptability and phenotypic plasticity of H . vulgaris ensured its adaptation to the environment . Also its tolerance contributed to the occupation of wider ecological amplitude .Second , the aggressive ability made it able to occupy the habitat in high density both ground and underground ,which may affect other species and led to a drop in community species diversity . Third ,it was apprehended that it had an extremely high reproductive speed , which may grow ５０ times each year , due to its vegetative propagation and seed propagation . At last , the difficulties in eradicating and the residue of its tissues also contributed to its rapid reproduction . The climatic standards to identify the potential distribution areas were defined as follows : the number of days whose daily average temperature was stably higher than (or equals) １０℃ were larger than １８０ d ; the lowest average monthly temperature (January) was higher than － ５ ℃ ; the annually extremely lowest climate temperature was not lower than － １０ ℃ . According to the meteorological data and climatic characteristics , north subtropical humid region , mid subtropical humid region , south subtropical humid region , edge tropical humid region , mid tropical humid region ,equatorial tropical region and plateau subtropical region were identified as the potential distribution areas of H . vulgaris in China . The invasion risk index assessment system indicated that H . vulgaris had a very high risk of invasion with the score of ７１ , which meant that H . vulgaris was extremely dangerous and was a forbidden species to be introduced .Some suggestions on invasion risk management were proposed on the basis of the growth habit of H . vulgaris .

A net pen experiment was carried out to evaluate the effect of feeding frequency on growth ,feed utilization and nitrogen and phosphorus waste output of malabar grouper , Ep inephelus malabaricus . In the experiment , fish [initial body mass ( １３７ ± １) g] were fed raw fish feed at ５ feeding frequencies , including ４ meals per day (４.００ meal.d^-１ ) , ２ meals per day (２.００ meal.d^-１ ) , １ meal per day (１.００ meal.d^-１ ) , １ meal every ２ days (０.５０ meal.d^-１) or １ meal every ４ days (０.２５ meal.d^-１ ) . The results indicated that feed intake ( FI) , mass gain , feed conversion ratio ( FCR) , nitrogen waste output ( TNW) and phosphorus waste output ( TPW) increased , while nitrogen retention efficiency (NRE) and phosphorus retention efficiency (PRE) decreased with the increase of feeding frequency from ０.２５ meal.d^-１ to １.００ meal.d^-１. However , no significant changes were found in FI , mass gain , FCR ,NRE , PRE , TNW and TPW ,when feeding frequency was further increased from １.００ meal.d^-１ to ４.００meal.d^-１ . There were no significant differences in condition factor , hepatosomatic index and whole body composition among fish fed at different feeding frequencies . The results revealed that the suitable feeding frequency for malabar grouper fed raw fish feed should be at １.００ meal.d^-１ . Fed at １.００ meal.d^-１ for １０９ days , mass gain of malabar grouper was ２５３ g , and FCR was １.１ , and NRE was ２２％ , and PRE was ２４％ , and TNW was １０８ g N.kg ^-1 fish production , and TPW was ２４ g P.kg ^-1 fish production .

To understand the effects of biochar application on properties of red soil , a pot experiment was conducted to study the influences of different application rates of biochar on soil quality indexes of two red soils varying in their fertility levels . The effect of biochar application on ryegrass growth was also observed . The results showed that the application of biochar to the red soils not only increased greatly storage of organic carbon of the soils , but also increased pH value , base saturation , cation exchange capacity , and amounts of water‐stable aggregates , field moisture capacity , available N , P and K of the soils , and decreased acidity and bulk density of the soils , and thus modified growth environments of ryegrass , and improved its growth . As compared to the control , concentrations of organic C , available P , K and base saturation of biochar‐treated soils increased by ３１％--７４４％ ,１４％--２１５％ ,６％--１１０％ , and １７％--８２％ , respectively , and soil pH increased by ０ .１１--０ .４０ unit ,at the application of biochar of １０ and ５０ g.kg ^-1 . The effects of biochar application were related to fertility levels of the soils ,and were greater in the low‐fertility soil than in the high fertility soil . The modified effects in the low fertility soil increased with increasing the rates of biochar application . However ,high application rate of biochar ( ２００ g.kg ^-1 ) might result in decline of micro‐biomass C in the high fertility soil , and limited slightly the growth of ryegrass .

By using sand culture , the changes of physiological and anatomical features and the characteristics of magnesium (Mg) distribution of Citrus cultivars shiranuhi" [ Citrus reticulata × ( C . reticulata × C .sinenesis)] and ?ponkan" ( C . reticulata) seedlings were studied in the Mg-deficient conditions . The results showed : １) The deficiency of Mg decreased significantly the plant biomass , root viability , leaf chlorophyll contents and superoxidase dismutase ( SOD) activities of both cultivars , especially for shiranuhi" . Mg deficiency increased significantly peroxidase ( POD) activities , malondialdehyde (MDA) contents and relative conductivity of both plant leaves . Furthermore , MDA contents and relative conductivity in shiranuhi" were significantly higher than those in ponkan" . In addition , leaf catalase (CAT ) activity of shiranuhi" in Mgdeficient
condition increased significantly , but it wasn't observed on ponkan" . ２ ) Mg contents of both cultivars leaves increased with increasing Mg concentration in solution , and they decreased obviously with the leaf aged . Furthermore ,Mg contents of shiranuhi" were significantly higher than that of ponkan" under Mgdeficient condition . ３) Ultrastructural observation of leaf structure and chloroplast showed that the organelle of shiranuhi" was damaged seriously under Mg-deficient condition , which was revealed by the vague thylakoid lamella with less number ,drastic increase of starch grain volume ,and the blurred membranes of chloroplast and mitochondria .As to ponkan" ,distortion ,plasmolysis and less grana lamella of chloroplast were observed ,but the starch grain and plastoglobuli numbers were increased . Thus , it is suggested that the influences of Mg deficiency on physiological and anatomical features were more obvious in shiranuhi" than in ponkan" .

To study the application of quantum‐inspired evolutionary algorithm (QEA) in the analysis of near infrared ( NIR) diffuse transmission spectroscopy of apples , first , spectroscopy regions were preselected by using the backward interval partial least squares (BiPLS) . Second , the variables were selected with QEA , and QEA‐PLS model was built . Meanwhile , genetic algorithm ( GA) -PLS model was developed to contrast with QEA‐PLS model . After running GA and QEA １０ times separately , the
two best models were chosen from the １０ GA-PLS models and the １０ QEA‐PLS models . The results showed that the GA‐PLS model had １１０ variables , with RMSEC ( root mean standard error of calibration) of ０.５８２ ０ , RMSEP (root mean standard error of prediction) of ０.６１２ ３ , but the QEA‐PLS model had １９４ variables ,with RMSEC of ０.４９２ ７ , RMSEP of ０.５２６ ０ . It is concluded that QEA can be used in the analysis of NIR diffuse transmission spectroscopy of apples and enhance the precision of model . Compared to GA , search capability of QEA is better .

A new kind of wastewater treatment system which used soil as the main substrate , multi-soillayering system , has been developed in Japan . It is composed of soil mixture block layers arranged in a brick-like pattern and water‐permeable layer .It has many advantages compared with other land treatment systems , such as preventing clogging , increasing hydraulic loading rate , enhancing the purification ability of soil , et al . This paper summarized the structure , mechanism and application of multi‐soillayering system , to provide a reference for the future use of multi-soil-layering system in China .

Remediation efficiency of cadmium (Cd) and benzo [a] pyrene ( B[a]P) combined polluted soil by Sedum al f redii Hance was studied using pot experiment under controlled temperature and natural light during cultivation .The results showed that the growth of hyperaccumulating ecotype ( HE) of S .al f redii was inhibited under high B[a]P level (2 5mg/kg ^-1 ) , whereas the biomass of nonhyperaccumulating ecotype (NHE) decreased significantly due to Cd toxicity . When treated with low Cd (1 mg/kg ^-1 ) , the root biomass of both HE and NHE were increased by 5mg/kg ^-1 B[a]P . Cadmium concentration in the shoot of the HE was much greater than the NHE and was not influenced by B[a]P supply level . The B[a]P concentrations in both shoot and root of both ecotype of S . alfredii were controlled by B[a]P level in the soil instead of Cd . The removal efficiency of Cd from Cd‐B[a]P combined polluted soil by the HE was much greater than the NHE , however , significant decrease of Cd removal efficiency was observed with treatment of 25mg/kg ^-1B[a]P . The removal efficiency of B[a]P from unplanted soil was43.7%, whereas it was increased to 58.2% after planted with both ecotypes of S . alfredii .All the results above suggest that hyperaccumulating ecotype of S . al f redii has great potential for phytoremediation of Cd and B[a]P moderately contaminated soil .