A couple of primers was designed on the base of VP２gene sequence of infectious bursal disease virus (IBDV) HZ２strain , and the VP２gene was successfully cloned into eukaryotic expression vector pDsRed２‐N１ . Then , the recombinant plasmid pDsRed２‐N１‐VP２ was constructed . The ２７９site and ２８４site on VP２ protein were mutated singly or homeochronously through site‐directed mutagenesis on theconstructed vector pDsRed２‐N１‐VP２ . By this way , three mutants of P２７９ , P２８４ and PDA were constructed in next step . The Vero cell line stably expressing VP２ protein and this mutant were also constructed successfully on the base of P２７９ , P２８４ and PDA by indirect immunofluorescence test . Therecombination plasmid and cell line constructed lay the foundation on the deep research about the role and mechanism of ２７９ site and ２８４ site in virulence change and antigenic variation of IBDV , which also play a significant role in exploring more efficient and cheaper IBDV DNA vaccination .

In order to develop novel molecular markers for faba bean ( V icia f aba L.) , the transferability of pea ( Pisum sativum L.) EST‐SSR (expressed sequence tag‐simple sequence repeats) markers and their application were studied in faba bean germplasms.The results revealed that ９９of １６３‐pair primers for pea EST SSRsachieved the effective amplification in faba bean , indicating a significant transferability ( ６０.７４％ ) of pea ESTSSR primers to faba bean.Thirty‐six of these transferable markers exhibited polymorphism and totally １４８alleles were detected with an average variation of ４.１.The polymorphism information contents of polymorphic markers varied from ０.０３５to ０.８１０ with an average of ０.４８３４, while observed heterozygosity ( HO ) ranged from ０.０００１to ０.７０００and expected heterozygosity ( HE ) ranged from ０.０３５７to ０.８４５２,with an average of ０.２１０３and ０.５３８８respectively.Furthermore , UPGMA ( unweight pair group method with arithemetic
averages) analysis and principal coordinate analysis (PCA) revealed that the ３０germplasms were classified into two major clusters.Because of their polymorphism and transferability , the developed markers in this study will provide the valuable tool for genetic diversity , resource conservation , comparative mapping
and marker‐assisted breeding of faba bean in the future.

Single nucleotide polymorphism ( SNP) is a kind of DNA polymorphism in genome which results from the variance of single nucleotide . In diploid organisms ( such as human , A rabidopsis thaliana , Oryz a sativ a , etc .) , many SNP markers were discovered as genetic markers ,and had been widely used for population parameter estimation and association mapping .However , SNP discovery and application in the polyploid organisms ( such as Gossy p ium hirsutum , Brassica napus , T riticum aestivum , etc .) were limited and lagged . In order to facilitate SNP discovery in polyploid plants , this paper reviewed the genomic complexity of polyploidy , the obstacle and the solutions of SNPs discovery in polyploid plants , including these methods of sequencing of polymerase chain reaction ( PCR) amplicons from locus‐specific primer , distinguishing SNPs from homoeologous sequence variants ( HSVs) through living models of allopolyploid ancestral genomes , use of next‐generation sequencing for SNP discovery ,scanning along seeking potential SNPs by bioinformatics tool , Mendelian transmission test of candidate SNPs . In conclusion , the applications of solutions mentioned above are feasible and reliable for the highthroughput SNP discovery in polyploid plants.

Base on the data for three dimensional ( ３D) coordinate of ４９neurons , １６morphological parameters were obtained by calculation . According to the factor analysis of the morphological parameters , three major characteristics , including dimension , degree of divergence and growths were obtained and chosen for classification . ward?s method was applied in the system cluster analysis . The results showed that the neurons could be classified into ５grades , including motoneuron , bipolar
interneuron , cone neuron , multipolar interneuron and tripolar interneuron , when the Euclidean distance was about １.５ ; additionally , the development degree of neurons affected the classification results of sensory neuron , tripolar interneuron and multipolar interneuron morphology , therefore ,in the process of
classification in morphology , it should be used mature neurons to avoid to cause interference.

Photo‐physiological mechanisms of tolerant difference to high temperature between two varieties of Celosia cristata , 'Century green' and 'Century copper 'were investigated . The parameters of gas exchange , chlorophyll content , light energy utilization and distribution were measured . The results showed that , 'Century green'had higher chlorophyll a (chla) content , which made more energy distribute to quantum yield of photosystem Ⅱ ( ΦPS Ⅱ ) . Meanwhile , higher non‐photochemical
luorescence quenching associated with state transitions ( qT ) in 'Century green'helped to dissipate excessive light energy timely . 'Century copper'had a higher tolerance to high light , but photoinhibition happened under high temperature for non‐stomatal factors , which reduced light energy utilization .Meanwhile , 'Century copper'can not dissipate excessive light energy because of lower chla content and lower proportion of qT . As a result , 'Century copper'was more sensitive to high temperature than 'Century green'.

In order to study the physiological characteristics of three typical Phalaenopsis spp . varieties (cold‐tolerant variety 'Hunyan' cold‐susceptible variety 'ubao', medium cold‐tolerant variety 'ZD‐１') under low temperature stress , the activities of catalase (CAT) , superoxide dismutase (SOD) and ascorbate peroxidase (APX) , relative electrical conductivity , soluble sugar and malondialdehyde (MDA) contents in leaves of Phalaenopsis spp . were determined at １０℃ , ５℃ , ０℃ for １２, ２４, ４８h after pretreatment. The results showed that the relative electrical conductivity and MDA contents in leaves were enhanced along with the increase of low temperature stress . Therefore , these two physiological indexes could be used as identification indexes of cold tolerance for Phalaenopsis plant . No obvious changes of soluble sugar contents were detected . The activities of CAT and SOD increased from １２h to ２４h and decreased from ２４h to ４８h at all low temperature treatments . The activity of APX consistently increased with the increase of stress degree . The activity of APX could also be used as an identification index of cold tolerance .It was found that the variety ?Hunyan?with good tolerance to low temperature possessed self‐protection mechanism through enzyme defense systems , while the susceptible variety 'Jubao'had low protection ability to cold stress .

Using total dietary fiber ( TDF) as index , optimization of fermentation process from wheat bran by lignin‐degrading fungus １６‐２ was studied ,and the characters of dietary fiber product obtained under the optimum condition were measured.The results showed that the optimum condition was pH ６.０ , fermentation temperature of ２８℃ , and fermentation time of ７２h.Under the optimum condition ,the total dietary fiber ( TDF) , insoluble dietary fiber ( IDF) and soluble dietary fiber ( SDF) contents
were ８２.４１％ , ５５.１２％ , and ２７.６０％ , respectively.Expansibility and water binding capacity of the product were ０ mL.g ^{－ １} and ４.２ g.g ^{－ １} , respectively.The contents of the cellulose , lignin ,crude protein and ash in dietary fiber were ３０.１２％ , １８.２８％ , ７.３９％ and １５.０４％ , respectively.The maximum binding capacities of the dietary fiber for Cd２ ＋ , Pb２ ＋ , Cu２ ＋ were ６６.９ , ２４９.６ , and １５.９ μmol.g ^{－ １} , respectively , and the removal ability for hydrogen peroxide was ２３.１％ .As a result , the water bindingcapacity and physicochemical activity of dietary fiber from the wheat bran obtained by microbialfermentation were good.

One strain , named as Ab , which was isolated from Oxytropis kansuensis Bunge and could degrade locoweed toxin — swainsonine , was screened by selective enrichment culture when swainsonine was used as a sole carbon source . Its degradation rate could reach １１ .５７７ １％ under the circumstance of being incubated on a shaker at １５０r.min－１and temperature at ２８℃ within ３０hours when the concentration of swainsonine was １００mg.L － １ . The capability of degrading swainsonine could not be improved by extending cultivation time , while it could tolerate high concentration swainsonine up to １ g.L － １ . Strain Ab was identified by １６S rDNA sequencing , morphological , biochemical and physiological experiments . The results showed that strain Ab was gram‐negative bacteria whose colonies were round ,milk white ,glossy and had raised‐center and regular edge on the Luria‐Bertani plate . And the colony was Brevundimonas and shared the highest homology ( １００％ ) with Brevundimonas diminuta by Blast software analysis . It is concluded that one strain of endophytic bacterium for degrading swainsonine , B .diminuta strain Ab , was existed in O . kansuensis . Therefore ,it is speculated that endophytic bacteria of degrading swainsonine might keep the homeostasis of swainsonine together with endophytic fungi of producing swainsonine in locoweeds .

In order to reduce consumption of sawdust , make full use of the resourceful corn stalks in Lentinula edodes ( Berk) Pegler cultivation , five indexes including the contents of carbon ( C ) and nitrogen ( N ) , C/N ratio in cultural materials , the contents of essential amino acid ( EAA ) and nutritional index ( NI) in L.edodes were analyzed and optimized by formula uniform design with corn stalk powder as main cultivation substrate , sawdust and wheat bran as supplemental cultivation substrates.The results indicated that the contents of corn stalk powder and sawdust was significant negative and positive correlation with the content of EAA and NI , respectively . Moreover , the contents of C and N , C/N ratio were regressive correlation with EAA , and prominent positive correlation with NI.Therefore , there was not a pronounced change in the contents of EAA and NI when the content of corn stalk powder was lower than ７４.５０％ and the content of sawdust not less than １１.６４％ , indicating that it was feasible to cultivate L . edodes with corn stalk instead of sawdust with important economic and social values .

The litterfalls of evergreen broad‐leaved forest in Baishanzu Mountain were collected by seed rain collector , and they were classified , identified and weighed.The litterfall composition and the monthly dynamics of leaf litter in the forest were also analyzed.The results show that : １) The amount of the litterfall in the forest was ４２４７.４７ kg.hm－２.a－１, with leaf litterfall of ７１.５４％ ( evergreen leaf litterfall of ５１.３４ ％ , deciduous leaf litterfall of ２０.２０％ ) , tree bark and branches of ３.１７％ , reproductive organs of １９.２９％ , debris of ６.００％ , respectively ; ２ ) The nine dominant species of litterfall had a clear leaf litter dynamics , and the evergreen trees could be classified into three types as follows : the deciduous trees had a peak of leaf litterfall in the autumn and winter ,and the evergreen trees had a peak of leaf litterfall in the summer , and the evergreen trees had a peak of leaf litterfall in the summer , autumn and winter ; ３ ) Compared with those of evergreen broad‐leaved forest located at the similar latitude , the litterfalls of the Baishanzu Mountain forest were much less , which was a composite reflection for the certain zones of the environment ; ４) Flowering and fruiting had a great effect on thedynamics of the litterfalls.Besides , typhoon played an important role on them.

Using maize hybrids Shenyu ２１and Dongdan ９０as entries , the effects of magnesium fertilizer on photosystem Ⅱ ( PS Ⅱ ) chlorophyll fluorescence characteristics of maize were studied through measuring chlorophyll fluorescence parameters such as ABS/RC , ET _０ /RC , TR_０ /RC , M_０ , Sm and φE_０ in six magnesium fertilizer levels (０ ,２２.５,４５,６７.５,９０, and １１２.５kg.hm^{－ ２} ).The results showed that , compared with the control , the chlorophyll fluorescence parameters in Shenyu ２１ increased significantly or highly significantly when the magnesium fertilizer amounts ranged from ４５ to ６７.５ kg.hm^{－ ２ .}The magnesium fertilizer did not affect or decreased the parameters beside φE_０ when the fertilizer level was more than ６７.５ kg .hm^{－ ２} .As for Dongdan ９０ , the chlorophyll fluorescence parameters increased significantly or highly significantly at the application amounts of magnesium fertilizer between ６７.５and ９０.０kg.hm^{－ ２,} but changed less beside the parameter of TR_０ /RC when the fertilizer amount was less than ６７.５ kg.hm^{－ ２} ,compared with control . Therefore , the PS Ⅱ linear electron transfer efficiency couldbe promoted , and assimilatory ability could be increased , and light utilization efficiency could be enhanced under the magnesium fertilizer levels of ４５‐６７.５ kg.hm － ２ in Shenyu ２１or ６７.５‐９０ kg.hm^{－ ２} in Dongdan ９０.

A pot experiment was conducted to explore the effects of applying organic liquid fertilizer made by coenzyme Q１０ fermentation broth on pakchoi growth and soil enzyme activities.The results showed that the chlorophyll contents , nitrate reductase and root activities of pakchoi were increased , and the nitrate contents were decreased by applying the organic liquid fertilizer ,compared with the control (basal fertilizer).Compared with the control , the biomass of pakchoi was increased by ２.０７％ ‐１４.５８％ bypplying the organic liquid fertilizer , and especially spraying the fertilizer diluted to ３００times had thebest effects.Significant and highly significant positive correlations were found among biomass , nutrientcontents and apart of physiological indexes.The activities of urease , catalase , sucrase were improvedsignificantly by pouring the organic liquid fertilizer , thus increasing soil available nutrient contents and fertilizer utilization efficiency , and promoting pakchoi growth.

In order to adjust to the peach structure ,extend the supply period and choose suitable varieties for production area in Zhejiang Province , six middle and late maturity honey cultivars , including'Qingshuibaitao', ;Qiubaitao', 'Huangjintao'and 'Xinchuanzhongdaobaitao'introduced from Japan ,and other two Chinese cultivars 'Qiuyulu'and 'Xumi'were evaluated , based on their size , mass ,firmness , total soluble solids , titratable acid , soluble sugar , organic acid and key aroma components .Two main varieties of yellow‐fleshed 'Jinxiuhuangtao'and white‐fleshed 'Fenghuayulu'were used as control . Meanwhile , the biological characteristics of each cultivar also were commented during the flowering and fruit development periods . The results showed that 'Huangjintao'was better than 'Jinxiuhuangtao'in flavor qualities , which had the higher soluble solids , total aroma content and lower titration acids , however , the former maybe experience late‐frost at the flowering stage . Three whitefleshed cultivars from Japan were better in the fruit mass , total soluble solids / acid ratio and storage character . Especially , the fragrance of 'Qingshuibaitao'was the strongest . Two domestic cultivars 'Xumi'and 'Qiuyulu'had obvious advantages in the fruit mass and storage characteristic compared with the cultivar 'Fenghuayulu', and other flavor qualities had no significant differences . However , 'Xumi' was more coarse in texture .

The degree of fatty liver in different color livers for edible grass carp ( Ctenopharyngodon idella) was discussed.Liver moisture was determined by freeze‐drying method , and liver crude fat was measured gravimetrically after ether extract using Soxhlet method.The hepatocyte lipid droplets were observed by light microscopy in oil red Ostaining for frozen sections.Based on the criterion of human'fatty liver , the degree of fatty liver in edible grass carp was then determined by the positive area ratio.
Compared with normal livers of grass carp , the moisture contents of motley livers , blushing livers and yellowing livers were decreased by ６.３０％ ,１０.０１％ and １４.１７％ ( P＜ ０.０５) , respectively ; the crude fat contents were increased by １９８.２５％ , ３３１.００％ and ４８５.５９％ ( P＜ ０.０５) , respectively ; the number of
hepatocyte lipid droplets were significantly increased with the peak in yellowing livers , and the positive area ratios were ３.９２ ,６.３１ and ７.３２ times of the normal liver ( P＜ ０.０５) , respectively.The degree of hepatocyte fatty degeneration in normal liver ,motley liver ,blushing liver and yellowing liver was １.１０ ±０.３０ , ２.３０ ± ０.２０ , ３.４０ ± ０.５０ and ４.００ ± ０.２０ , respectively.After doing linear regression analysis ,liver fat contents ( Y ) were negatively correlated with its moisture contents ( X ) : Y ＝ － １.０８９ ９ X ＋８２.０３５ ７ ,R２ ＝ ０.９８９ ( P＜ ０.０１) ; the positive area ratio ( Y ) were positively correlated with its crude fat content ( X) : Y ＝ ０.９７０ ２ X － ０.２０９ ０ ,R２ ＝ ０.９７３ ( P＜ ０.０５) ; the crude fat content (Y ) were positively correlated with the degree of hepatocyte fatty degeneration ( X ) : Y ＝ ３.６６２ ４ X － １.７８８ ６ ,R２ ＝ ０.９８５( P＜ ０.０１ ) ; the positive area ratios ( Y ) were also positively correlated with the degree of hepatocyte fatty degeneration ( X) : Y ＝ ３.６２２ ４ X － ２.１３０ ６ ,R２ ＝ ０.９９６ ( P＜ ０.０１).These results confirm that the oil red O staining method for frozen sections is suitable for evaluating the degree of fish fatty liver.As aresult , according to the positive correlation between the positive area ratio and the crude fat content or the degree of hepatocyte fatty degeneration,it is suggested that the liver in grass carp can be defined as fatty liver when its positive area ratio exceeds ２％.Correspondingly , the degree of fatty liver can be defined as mild when its positive area ratio ranges from ２％t o７％,as medium when ranging from ７％to １２％, and as serious when exceeding １２％ .

PCR‐terminal restriction fragment length polymorphisms ( PCR‐TRFLP) combined with the method of １６S rRNA gene clone library building and sequencing were used to investigate the changes in the bacterial communities of tray‐packaged pork during ４℃ storage . The results showed that ３５terminal restriction fragments ( TRF ) were observed in the TRFLP profiles , indicating that the composition of bacteria population in chilled pork was complex . The peak of the TRF corresponding to Pseudomonas varied significantly during the whole storage , and the relative peak area continued to increase from ５.７％ at day ０to ８３.４％ at the end of monitoring , which indicated that Pseudomonasbecame the predominant bacterial species in the end .

Fuzzy mathematic evaluation model was introduced to optimize the process of stewed pork . As five key factors , colour , texture and mouthfeel of the stewed pork , taste and odour of the soup were on weight analysis based on sensory evaluation , and the integrated assessment system was established . On this basis , different ratios of material to water , stewed temperatures and time were optimized by the method of fuzzy mathematic evaluation . The results showed that the optimum stewed conditions were as follows : the pork to water ratio of １ ∶ １ .５, stewed temperature of ７０℃ and stewed time of ２h .Sensory evaluation combined with fuzzy mathematics make the estimation more scientific , reasonable and objective , which will have broad application perspectives in the field of food processing .

The main heavy metals and pesticide residues of １０kinds of vegetables for sell in １１prefecture cities of Zhejiang Province were monitored and evaluated . The results showed that １７kinds of pollutantsdetected were : cadmium ＞lead ＞cypermethrin ＞mercury ＞chlorpyrifos ＞cyhalothrin ＞triazophos ＞acephate ＞acephatemet ＞ fenvalerate ＞ folimat ＝ parathion ＝ malathion ＝dichlorvos ＝ fenpropathrin ＞ dipterex ＝deltamethrin ; the indexes of high risk were : cadmium ,chlorpyrifos , lead , cypermethrin , triazophos , and the indexes of secondary risk were : acephatemet ,cyhalothrin , folimat , parathion , and the remain indexes were low risk ; the indexes for exceeding vegetable safety acceptable limit were : cadmium , triazophos ,and acceptable limit indexes were : folimat ,acephatemet , lead , and the remain indexes had no effect on vegetable safety ; safety indexes of different kinds of vegetables were all less than １ , indicating that the total circumstance was well ; safe level of vegetable from high to low was : cucumber＞tomato＞suakwa ＞cabbage ＞water bamboo ＞celery ＞leek ＞Chinese cabbage ＞eggplant ＞cowpea ; safety indexes of vegetables in different subregions were all less than １ ,and safe level from high to low was : Hangzhou ＞Jinhua ＞Shaoxing ＞Wenzhou ＞Lishui ＞Huzhou ＞Taizhou ＞Zhoushan ＞Ningbo ＞Jiaxing ＞Quzhou .