In order to extract celery CEL I nuclease effectively and use it to develop the detection method for point mutations , the effects of varieties ,organs ,developmental stages and pH of extraction buffer on its extraction amounts were analyzed and its enzymatic activity for mismatch cleavage was also characterized by using DNA heteroduplexes containing a single mismatch as substrates . Results showed that differences of CEL I amounts extracted among varieties and organs were significant at ５％and １％level , respectively . The cultivar Thailand Huangxinqinshowed the highest extractive amount among three different varieties ( Xuebai, Wentula and Thailand Huangxinqin ) compared , while the leaf showed much higher extraction amounts than the stalk and root among three different organs tested . The effect of developmental stages on the CEL I extraction was compared by using the variety Shanghai Huangxinqin as extractive material and it was higher at reproductive stage than that at vegetative stage . The pH of extraction buffer also showed important effect on the CEL I extraction , and the higher extraction amount was obtained by using the extraction buffer with pH ７.５ than that with pH ５.５ andpH ９.５. Tests on enzymatic activity showed that the mismatch in DNA heteroduplexes could be effectively recognized and cleaved by the CEL I crude extracts .

Three identification methods for the resistance to Sclerotinia sclerotiorum previously‐reported in Brassicaceae crops were compared using ５cauliflower varieties under the same condition to develop the effective screening technique in cauliflower . The results showed that the correlation between inoculating mycelium agar disk to leaf axil at seedling stage ,or inoculating mycelium agar disk to live leaf at heading stage and field inoculation were highly significant ( r ＝０.８８５９and ０.９２６６, respectively) , so these two methods could well identify resistance and susceptibility of cauliflower varieties . The method of inoculating mycelium agar disk to in v itro seedling leaf was not suitable for resistance identification of cauliflower . To screen resistant sources of cauliflower against S . sclerotiorum , ２１ breeding materials were tested with the method of inoculating mycelium agar disk to leaf axil at heading stage , using cauliflower variety nongle ４５ days as control . The results showed that there were two high resistant materials , shenliang golden flower and changsheng ６５days .

To investigate molecular‐assisted breeding in M iichthys miiuy , a normalized full‐length cDNA library was established using the duplex‐specific nuclease (DSN) normalization method combined with switching mechanism at ５ end of RNA transcript (SMART) technique . The titer of un‐amplified cDNA library was about １.６ × １０^６CFU.mL ^{－ １} ; the cDNA library contained ７.５ × １０^６ independent clones with DNA inserts of １‐３ kb . A total of ５９５２clones were randomly selected to be sequenced and ５０５３expressed sequence tags ( EST) were obtained , and ４６０９ high quality EST sequences were generated .After processing , a total of ３２２１ unigenes comprising ６５６ contigs and ２５６５ singlets were obtained , and the redundancy ratio was about ３０.１１％. The average reading length was ５５０bp , and reading length of most sequences was ranged from ５００to ５９９bp and the GC contents of most sequences was ranged from４０to ６０percentages . A total of１３９７ functional known genes were categorized by gene ontology . Many putative immune genes were identified that encoded major histocompatibility complex , cytokines and cytokine receptors , adhesive proteins , stress proteins , transcription factors for immune response ,immunoglobulin and coagulation factors . The results above provide both a detailed annotation of immune genes and a collection of novel transcripts for this important species .

In order to provide some references for studying the genetic variation and selecting varieties of Tenebrio molitor , the esterase isozyme zymograms and activity at different developmental stages of the yellow‐ and black‐color varieties of T . molitor were compared using vertical polyacrylamide gel electrophoresis and spectrophotometry . The results showed that １５ enzyme bands with different mobility were found at all , which were distributed in five zones . The larvae and adults of the black‐color variety
showed one more distinctive band compared with those of the yellow larvae and adults , respectively . Moreover , band numbers , migration rate of enzyme bands and strength of enzymatic activity at differentdevelopmental stages of both color varieties varied greatly . Compared with other developmental stage ,band numbers of esterase isozyme were the least ( only four bands) and enzymatic activity was the weakest at egg stage , and two of four bands were found only at this stage . The enzymatic activities of carboxylesterase ( CarE ) , acetylcholine esterase ( AchE ) , acid phosphatase ( ACP ) and alkaline phosphatase (AKP) in black‐color variety were always higher than those in yellow‐color variety at all stages from egg to adult . While in the same variety , The enzymatic activity of CarE , AchE , ACP and AKP varied in the order of egg stage ＜ adult stage ＜ pupal stage ＜ larval stage . The results above build up some underlying data for further studying the insecticide resistance and variety selection of the
mealworm beetles .

A novel bioflocculant‐producing Bacillus sp . was isolated from soil by streak plate method , and screened out by an index that the fermentation broth flocculated kaolin suspension . The strain was identified by physiological and biochemical characteristics and １６S rDNA sequence analysis . The effect of the factors such as fermentation broth dosage , Ca^{２ ＋} concentration and pH on the flocculation were investigated . The results revealed that a novel bioflocculant‐producing bacterium , named MBFF６ , was isolated from soil and identified as Bacillus megaterium ; its flocculating rate could achieve ９５.７％ ; the optimal condition for flocculation was as follows : ermentation broth dosage of ２％ , Ca^{２ ＋} concentration of ４.５ mmol.L ^{－ １} and pH ７ .０The bioflocculant was produced during the bacterial growth and was an extracellular product which was distributed in the culture broth.

The effects of heterotrophic transformation and autotrophic transformation on astaxanthin production in H aematococcus p luv ialis were studied and compared . When sodium acetate was added in MCM for heterotrophic transformation ,astaxanthin accumulation was increased from ０ .７２３ ６ mg.L － １ to １.２１６ ８ mgL ^{－ １} .To enhance the astaxanthin accumulation , single factor experiment was founded up.Center composite design experiments and response surface methodology were employed to optimize the media of heterotrophic transformation.The results showed a distinctive influence of sodium acetate and potassium nitrate concentrations on astaxanthin accumulation.The optimal fermentation medium consisted of １.４６０ ５ g.L ^{－ １} sodium acetate , ０.００８ ４ g.L ^{－ １} potassium nitrate , and ０.０２ g.L ^{－ １} potassium dihydrogen phosphate.Astaxanthin content reached １０.９０８ mg .L ^{－ １} , which was raised １０.１６‐fold against the control group and １４.０７‐fold against the group of autotrophic transformation.

In order to compare the effects of methanol , formaldehyde and formic acid application on plant physiological characters and gene expressions and to better understand the functional mechanisms of the three C１ compounds , ５ mmol.L ^{－ １} methanol , formaldehyde and formic acid were sprayed , respectively , on the leaves of A rabidopsis grown under greenhouse conditions once a week , using the plants sprayed with distilled water as the control . After four weeks , the effects of foliar spraying these C １ compoundson plant growth and physiological characters were analyzed . The results indicated that methanol stimulated the growth of A rabidopsis , whereas formaldehyde and formic acid inhibited its growth and the nhibition of formaldehyde was much remarkable . Application of methanol decreased anthocyanin content but increased chlorophyll ( Chl) a content and a/b ratio significantly . On the contrary , formaldehyde reatment reduced Chl a content and a/b ratio markedly . Application of formic acid caused an obvious increase in Chl a content and a notable decrease in Chl b content , which led to ２‐fold increase in Chl a/b ratio . Application of three C１ compounds all increased total leaf soluble protein content ,and the effect of methanol spraying was especially remarkable . Only formaldehyde treatment elevated the levels of soluble
sugars , H２ O２ and carbonyl‐proteins in leaves , suggesting occurrence of oxidative stress . Gene expression profiles by reverse transcription‐polymerase chain reaction (RT‐PCR) showed that methanol stimulation induced expressions of most selected photosynthesis‐related genes , while formaldehyde repressed their expressions but up‐regulated most of the selected stress‐responsive genes . The expressions of most selected photosynthesis‐related genes were not affected significantly by the application of formic acid . Interestingly , three C１ compounds had no remarkable impacts on the expressions of most selected genes involved in C１ metabolism , but all repressed the expression of ５ ,１０‐methylene‐tetrahydrofolate ( THF) reductase .

A series of pot experiments were conducted to understand the effects of salt stress on physiological‐biochemical characteristics and chloroplast submicroscopic structure of Rhododendron indicum . The results indicated a robust development of the individuals under low salt stress . The contents of soluble protein , catalase ( CAT ) , peroxidase ( POD) and superoxide dismutase ( SOD) activities were significantly higher than those of the control , while the variation of malondialdehyde (MDA) and chlorophyll concentrations were not significant . Submicroscopic observation suggested an unmodified structure of chloroplasts under low salt stress , and chloroplast grana lamella showed high level of regularity with not significant difference to that of the control . These results above indicates high fitness of R . indicum in the soil with relatively low NaCl concentration . However ,under １２０ mmol.L－ １ NaCl concentration , the leaves of R . indicum began to show mild symptoms , and illustrated a positive relationshipbetween the level of damage and the increment of salt concentration in soil . With the salt stress increased , the contents of soluble protein and chlorophyll decreased , while MDA content increased significantly , and POD , CAT and SOD activities increased first and decreased afterwards , and the chloroplasts showed an increase in dimension and became spherical in appearance , and the degree of cavity in chloroplasts increased with slow disintegration of their membrane , indicating that Rhododendron displayed certain level of salt resistance . But the structure and function of chloroplasts of R . indicum became increasingly damaged under high salt stress , which further led to the etiolation and senescence of its leaves .

Root‐zone restriction of half‐raised bed cultivation was applied to four grape varieties (Fujiminori, Kyoho, Summerblack and Jinshouzhi) in south China , and its effects on grape quality under protected cultivation were studied . The results showed that using the high‐density compacted soil of １５‐２０ cm below soil surface as a natural buffer zone , could well play the role of estricting root growth , and the root‐zone restriction could improve the grape inner , visual and nutrition quality . The ratio of total soluble sugar (TSS) to total acidity ( TA) was raised at the four varieties , and the anthocyanin and total phenolic contents increased at the three cultivars ( Fujiminori, Kyoho and Summerblack) by root‐zone restriction system . Besides the mass of Fujiminori berry was reduced significantly , that of the other three cultivars had not significant effect by root‐zone restriction of halfraised bed cultivation . These results suggest that the technique of root‐zone restriction of half‐raised bed cultivation in south China has a positive effect under facility cultivation condition .

Using ?Dongkui?bayberry cultivar as test plant , the effects of different antifreeze patterns on grain yield and fruit quality of bayberry grown in mountainous areas were studied in the field experiments during ２００８ and ２０１０ . Results showed that , compared with the control , the antifreeze patterns significantly reduced the decline rate of the nitrogen , phosphorus and potassium contents of bayberry leaves in winter , and the effect of the greenhouse pattern was the best , whose decline rate was １１.０％ ,２０.３％ , １６.５％ , respectively , and the water content of bayberry leaves under greenhouse pattern remained stable in the winter period . The antifreeze patterns could promote the growth of spring shoots and increased their nitrogen contents .Compared with the control , the increasing rate of nitrogen contentof spring shoots under greenhouse pattern was the biggest , increasing by １４ .６％ . The antifreeze patterns significantly increased the nitrogen , phosphorus and potassium contents of fruits , especially the greenhouse pattern being the best . Compared with the traditional antifreeze patterns , the greenhouse pattern significantly improved mass , soluble solid , sugar and vitamin C contents of the bayberry fruit , and reduced the acidity of fruit , and then improved fruit quality of bayberry . The yield of bayberry under
greenhouse pattern was the greatest and increased by ７８.４％ compared with the control .

The potential of malabar grouper ( Epinephelus malabaricus) in utilizing pet‐grade poultry byproduct meal (P‐PBM) and a blend (MM) which was comprised of ６５％ feed‐grade poultry by‐product meal , ２０％ feather meal and １５％ blood meal was examined in a １０‐week net pen experiment . Five isonitrogenous (４９％ crude protein) and isocaloric ( １９ MJ.kg ^{－ １} gross energy) diets were established .One diet was formulated to contain ５０.０％ herring meal as control ,and in the other four diets , ３０％ and ６０％ of the fish meal were replaced respectively by either incorporating P‐PBM at １５.０％ and ３０.０％ ,or incorporating MM at １３.９％ and ２７.８％ . Each diet treatment had three replicates . Initial body mass of the experimental fish was (３３.４ ± ０.１) g . The results indicated that reducing dietary fish meal level from ５０.０％ to ２０.０％ , with P‐PBM or MM as alternative protein source , had no significantly negative effect on feed intake , body mass gain , feed conversion ratio , nitrogen retention efficiency (NRE) , phosphorus retention efficiency ( PRE) and whole body composition . Partial replacement of fish meal with P ‐PBM
resulted in enhancement in body mass gain , NRE and PRE . The results above reveal that P‐PBM is an excellent dietary fish meal substitute for malabar grouper ,and using fish meal and P‐PBM in combinationas dietary protein source can enhance growth performance and feed utilization efficiency of malabar grouper reared in net pens .

An investigation was made to distinguish five rice varieties with PEN ２ electronic nose . The matched experiment factors including the sample mass , headspace volume and generated time were studied . The response signals of electronic nose (e‐nose) were analyzed in various sampling conditions by single‐factor analysis of variance . The results showed that the signals of e‐nose were stable under the condition with the sample mass of １０ g , headspace volume of ２００ mL and the generated time of ６０ min .Then the data were analyzed with principal component analysis method ( PCA) , linear discrimination analysis method (LDA) . The consistent results by LDA and PCA revealed that the grain and polished rice were superior to brown rice and cooked rice to identify at the same time . As a result , the different varieties of rice were classified precisely by e‐nose , which confirmed that e‐nose technique was a generaldetection for the comprehensive volatile substance with high content , thus it afforded the experimental data based on e‐nose technique for rice odor detection .

The essential oil from Perilla f rutescens were extracted by microwave method ,and the optimum extraction conditions were obtained through response surface methodology , then the constituents of the essential oil were identified by gas chromatography‐mass spectrometry ( GC‐MS) , in the meantime , with scanning electron microscope (SEM) , the damages of the cell structures were observed . The results showed that the optimum extraction condition was as follows : cyclohexane as solvent , microwave power of ３２９W , irradiation time of ８０s ,material to liquid ratio of １ ∶ ６ ,soaking time of ５６min . Under the optimum extraction condition , the actual detection value of the essential oil was １.７８３％ , that was almost equal to the predictable value .Seventeen compounds , identified by means of GC‐MS , made up ８０.６１％ of the essential oil , whose main components were perillaldehyde (４４.５４％ ) and D‐limonene (１５.７％ ) . Under the scanning electron microscope (SEM) , the cell wall had little damage during the irradiation , but the cell membrane and mitochondria were badly damaged ; although the outlines of several cellular organs were clear , including chloroplast and starch grain , they all had some contents effusing .

Mann‐Kendall approach was applied for characterizing the features and mechanisms of vegetation changes with time , elevation and slope in the Tiaoxi watershed , Taihu Lake region , Zhejiang Province , east China . The results indicated that the annual vegetation in the area fluctuated dramatically and showed a moderate trend of increase from ２０００to ２０１０, but in ２００４, the coverage dropped to the lowest level ; The variability of spatial distribution also showed a moderate trend of increase during the study period , with the spatial heterogeneity reaching the highest between ２００２and ２００４; the vegetation coverage kept increasing while the spatial heterogeneity decreased with the increment of elevation and slope . At elevation of about ２００ m or slope around １５ degrees , there were signs of catastrophe change .The results above reveal the trend of vegetation changes with time , elevation and slope , and identify when and where the catastrophes occur , which lays a sound basis for further study on the spatiotemporal dynamics of vegetation and its driving factors .

To solve the problem of subjectivity and arbitrariness , fuzzy weights of evidence method was introduced to a summer orange production areas for land suitability evaluation . Evaluation results showed that ８５％of the known appropriate points were in the revised medium‐appropriate level , indicating that the fuzzy weights which integrated data‐driven and knowledge‐driven , as long as combined with modified posterior probability model , could achieve an objective and accurate measurement for land suitability evaluation .