Abstract:Cellulose synthase is a critical catalytic subunit of the cellulose synthase complex.To study functions of CesA genes in rice,their antibodies were prepared.First,recombinant plasmids were constructed with cloning the cellulose synthase genes into the expression vector pGEX-4T-3.The recombinant plasmids were then transformed into E.coli JM109 and the recombinant proteins were obtained.The fusion proteins were purified with GST affinity chromatography system and injected to rabbit for generating polyclonal antibodies.Finally,all antibodies were identified by Western blotting analysis with specific bands and high activities.

Abstract:Ancymidol was investigated as a medium supplement for long-term storage of potato plantlets in vitro.Different concentrations of ancymidol (0,10,15,and 20 μmol/L) were evaluated in growth media based on MS medium with 80 g/L sucrose.The cultures of three genotype Solanum chacosense (cha),CE76 and E3 were preserved under a 16 h photoperiod at (17±1)℃.The results showed that ancymidol had an active effect on culture viability to prolong preservation in vitro.Different concentrations of ancymidol had diverse growth-inhibiting effects.After conserved in ancymidol media,potato plantlets did not show evident abnormal shapes like vitrification and flaccidity in the following subculture,which are very frequently observed under mannitol stress.Genetic stability of potato plantlets conserved in ancymidol media was analyzed with randomly amplified polymorphic DNA (RAPD) and inter-simple sequence repeat (ISSR) fingerprints.Ancymidol resulted in 0.48%-2.95% genetic variations in genomic DNA of different genotypes,which will motivate us to find out better concentrations of ancymidol supplied for long-term storage of different genotypes.

Abstract:Bacillus thuringiensis CT-43 produces thuringiensin.The VirD4 protein encoded by the virD4 gene located on the plasmid pBMB0558 of CT-43 is highly homologous to the coupling proteins (T4CPs) of type IV secretion systems.In type IV secretion systems,T4CPs’ basic function is to recruit substrates to the type IV secretion systems for secretion through the translocation channel.A homologous double - crossover vector was constructed through knocking out the virD4 gene in CT-43 and a mutant BMB1122 was obtained.HPLC result showed that the characteristic peak of thuringiensin was not detected in the fermentation supernatant of mutant BMB1122.The LCMS-IT-TOF result showed that the corresponding m/z with molecular weight of proposed precursors in Thu biosynthesis could be detected in intracellular metabolites of mutant BMB1122,consistent with that of CT-43.The knockout of virD4 gene interrupted the secretion of thuringiensin.Presumably,the virD4 was a critical gene for Thu biosynthesis.

Abstract:The antifungal substance produced by Bacillus amyloliquefaciens WH1,isolated from the root of rice,was identified with infrared spectra.The inhibitory effect of WH1 towards Colletotrichum gloeosporioides was measured by suppression tests both in the plate and leaves in vitro.The results showed that the antifungal substance WH1 produced was cyclic lipopeptide,the fermentation broth was detected with the activity to deform the mycelium of C.gloeosporioides,and the highly diluted fermentation broth still had a significant control effect in vitro.In order to make WH1 a dedicated formulation to control C.gloeosporioides,the fermentation condition of WH1 was optimized by Plackett-Burman design and Taguchi array design.After optimization,the lipopeptide production has increased significantly.Keeping the dissolved oxygen above 30%,the fermentation in fermentor of 3 L,20 L and 500 L was scaled up successfully.Tenfold diluted fermentation broth of fermentation in 500 L fermentor has inhibitory rate of 50.8% on C.gloeosporioides,and still kept inhibitory rate of 86.2% on fungus after spray drying.

Abstract:A new method for eradicating the double infected pathogen of CTV and CTLV incitrus was explored in this experiment.The citrus plants were selected from those conserved in net greenhouse of National Indoor Conservation Center of Virus-free Germplasms of Fruit Crops,Huazhong Agricultural University.The infection of pathogen was detected with reverse transcripts PCR (RT-PCR).Results showed that the suitable protocol was using the treatment of the antiviral medicament with Chinese medicines (eight times) and combinated with heat treatment of the highest temperature at 40℃ during the day(6 h/d),the temperature at 25℃ in the evening kept for fifty days in the PC made greenhouse.The protocol mentioned above was proved to completely eradicate double infected pathogen of CTV and CTLV; Furthermore,the combined treatment of shoot-tip grafting and the antiviral Chinese medicines was identified 85.7% eradication of the double pathogen.In above experiments,the leaves were sampled at 1,3,5,7,9 d after the Chinese medicines treatment.The results showed that the activity of the resistance related enzymes PPO,POD,SOD were increased compared with the control.

Abstract:The structural features of root,stolon,leaf and tuber of Vallisneria spinulosa were analyzed with plant anatomy.The results showed that Vallisneria spinulosa had structural characteristics of aquatic monocotyledon with well-developed adventitious root and multi-node stolon.The single grain starch granules were rich in the basic tissue of winter tuber.The aerenchyma is well developed,vascular structure is simple,xylem is degraded or absent and mechanical tissue is not developed in whole-plant.The Vallisneria spinulosa’s winter buds are proposed to be defined as “tubers”.

Abstract:The relationship of Pinus mussoniana radial growth with climate change in the upstream of Three-gorges Reservoir Area in China was assessed by dendrochronological techniques including correlation functions and single-years analysis.The results showed that Pinus mussoniana growth in the high and low elevations had a significantly different response to temperature and precipitation.In the low elevation,Pinus mussoniana growth was significant correlated with the mean precipitation and the humid indexes of of May and June.But in the high evelvation,it had a more complicated relationship with its environment.Besides the mean temperature of May and June,the humid index of last Novemeber was significantly correlated with the Pinus mussoniana tree-ring width.Therefore,the relationship of the same tree species with different environmental gradients was different.

Abstract:An optimal ISSR reaction system for Taiwania cryptomerioides was established with 25.0 μL reaction mixture made up of 40 ng genomic DNA,0.4 μmol/L primer,1.8 mmol/ L Mg2+,1.0 U Taq,0.2 mmol/L dNTP,and 2.5 μL 10×buffer.The PCR were performed as follows:4 min at 94℃,40 cycles of 45 s at 94℃,45 s at 51℃ to 56℃,and 2 min at 72℃,with a final extension of 7 min at 72℃.Ten polymorphic primers were screened using this reaction system.Twenty DNA samples from the wild population of T.cryptomerioides distributed in Lichuan City of Hubei Province were amplified by the 10 selected primers.The percentage of polymorphic bands was 61.97 %,Nei’s gene diversity and Shannon’s information measure were 0.136 9 and 0.216 0,respectively.

Abstract:Four isolates of the anthracnose pathogen were isolated from postharvest banana fruits displaying anthracnose symptoms by using tissue isolation technique. The differences in mycelial growth rate,conidial germination, appressorium formation and pathogenicity of the four isolates were compared under the same conditions. The results showed that there was no significant difference in the mycelial growth rate,but the percentages of conidial germination and appressorium formation, and the pathogenicity of isolate X4 were significantly higher than those of the other 3 isolates. The isolate X4 was identified as Colletotrichum musae based on the classical morphological taxonomy combined with the technique of ITS sequence analysis. Meanwhile, the effects of temperature, illumination and pH value on the mycelial growth, conidial germination and appressorium formation of isolate X4 were investigated. The results indicated that the temperatures for the mycelial growth were ranged from 10-35 ℃, with an optimum of 28 ℃ and the suitable temperatures for conidial germination and appressorium formation were from 25-30 ℃, with an optimum of 28 ℃. Illumination had no significant effect on the linear growth of mycelium, conidial germination and appressorium formation. The pH values for the mycelial growth and conidial germination were 5.0-8.0, with an optimum of pH 6.0. The lethal temperature of conidia and mycelia explored further were 50 ℃/10 min. 

Abstract:Resistance of the major commercial rape varieties in Yunnan Province and seven clubroot resistant rape varieties from other rape breeding institutions in China was studied in the Gaohai village, Banqiao town, Longyang region, Baoshan city in 2010 where the clubroot spread most extensively. The results showed that the Plasmodiophora brassicae were aggravated by artificial simulating waterlogging at seeding stage, while the yield was reduced greatly. The clubroot harmed rape from seedling period and was most severe at bolting stage while the disease index and disease incidence reduced at silique mature period. The sixteen rape varieties were divided into three classes according to their resistance to the clubroot. Tress varieties A35, Huayou No.7, Yunyoushuang No.1 showed high resistance during all growth period and could be used as dominant breeds for further study on resistant breeding.

Abstract:The morphology of antennae sensilla and sex pheromone gland of Maruca testulalis Geyer were observed under scanning electron microscope. Antennae were made up of scapus, pedicle and flagella. The latter consisted of 72-82 segments and was about 86-97 cm long. The upper surface of antennae was covered with cataphylla and most of the antennal sensilla lied on its side and lower surfaces. Six kinds of antennal sensilla were found in both sexes. Sensilla cylindric were observed on male antennae and an undefined sensilla was observed on female antennae only. Sensilla trichodeum of males were longer than that of females. The number of sensillum trichodeum was lesser on females antennae than males antennae. The study showed the sex pheromone gland was situated in the intersegmental membrane between the eighth and ninth abdominal segments, which was verified by GC-MS analysis.

Abstract:In this study, OBP from Bactrocera cucurbitae was cloned and named as BcucOBP. The full-length of open reading frame（ORF）is 447 bp, encoding 149 amino acid residues. Amino acid sequence alignment and 3D-Homology Modeling analysis showed that BcucOBP shared the typical structural feature of OBP with other insects, including 6 conservative Cys residues and 6α-helixes in the sequence. A recombinant plasmid pET28a (+)-BcucOBP was constructed and transformed into Escherichia coli BL21（DE3）. Western blotting indicated that the His-tagged BcucOBP stably expressed in DE3 induced by IPTG. Real-time PCR analysis shows that BcucOBP mRNA is expressed nearly in all tissues except for female visceral segment. The relative concentration of BcucOBP mRNA is the highest in antenna of B.cucurbitae compared with other tissues. BcucOBP mRNA shows clear gender-differential expression pattern in general section and the foreleg. The special expression pattern suggested that BcucOBP not only acted as a general odor protein but also was involved in the transport of pheromone in the mating behavior of B.cucurbitae. 

Abstract:The purpose of this paper was to evaluate the importance of weed control by 75% halosulfuron-methyl WG in the farming management of tomatoes was fields. The effect of 75% halosulfuron-methyl WG on the main weed control and the subsequent light penetration rate(LPR) at different heights of fields,as well as the yields of tomato were studied by means of field experiments and instrument mensuration. The results showed that:75% halosulfuron-methyl WG had significant control effects on the main broad leaf weeds and Cyperus weeds. The overall control effects of Cyperus rotundus,Amaranthus retroflexus, Melochia corchorifolia,and Acalypha australis were from 82.52% to 95.78%,and the overall quantity control effects were from 83.53% to 96.09%. The LPR in the middle and bottom of the tomato fields was significantly improved after the weed control. With weed control,the situation of light penetration rate was markedly improved,which resulted in a prominent enhancement of rice yield.

Abstract:To date,few studies have addressed the relationship of clonality and invasiveness of alien invasive plants in large geographical scale.The 112 alien invasive plants in North Carolina of America were analyzed.The results showed that the percentage of the strongest invasiveness plants,the stronger invasiveness plants,the weaker invasiveness plants in all invasive plants are 24.1%,44.6% and 31.3%.There are 67 invasive clonal plants (59.8%),whose mainly reproductive types were stolon and tiller.The percentages of annual or biennial herbs,perennial herbs,aquatic herbs,tree,shrub,vine and bamboo in total invasive plants are 23.2 %,20.5%,8.9%,7.1%,23.2%,15.2% and 1.8%,with clonal plants accounting for 46.2%,82.6%,100%,62.5%,34.6%,58.8% and 100%,respectively.With the increase of invasiveness,the percentages of invasive plant originated from Asia are 45.7%,66% and 74.1% in three ranks,and those originated from Europe are 45.7%,30% and 14.8%.Data mentioned above indicated that clonality of the invasive plants in North Carolina may be contributed significantly to their invasiveness,and Asia and Europe were the primarily geographical origin of these alien invasive plants.

Abstract:The antagonistic activity of an endophytic bacterium BYG2-5 against Colletotrichum musae was studied on cultural medium.The results showed that strain BYG2-5 significantly inhibited the mycelial growth of C.musae with the inhibition zone reaching up to 18 mm.The fermentation fluid and its filtrate showed a high efficacy in suppression of infection caused by C.musae on banana fruits.Especially,treatment effect of the fermentation fluid was 80.6% against C.musae. The strain was identified as Bacillus litoralis based on its morphology,physiological biochemistry and 16S rDNA sequence.

Abstract:Runoff plots experiment was conducted to study the effects of recommended fertilization,recommended fertilization with split-applied fertilization and recommended fertilization with planting grass on the output of runoff,nitrogen and phosphorus,form of nitrogen and phosphorus.Results showed that recommended fertilization with planting grass and recommended fertilization with split-applied fertilization significantly reduced runoff losses 38.43% and 26.51% compared with the conventional fertilization.Rainfall intensity is the main driving factor for the loss of nitrogen and phosphorus with more than 96% of nitrogen lost and 98% of phosphorus lost happened during the heavy rain and torrential rain.Dissolved nitrogen and particle phosphorus other than fertilizer application were the main loss forms of nitrogen and phosphorus in Danjiangkou Reservoir orchard.The loss of total nitrogen,dissolved total nitrogen and dissolved total phosphorus were deceased significantly by increasing the frequency of fertilizer application,and planting grass can effectively reduce the loss of total phosphorus and particle phosphorus.The loss of 35.96% total nitrogen was decreased by increasing the frequency of fertilizer application while the loss of 59.31% total phosphorus were reduced significantly by planting grass,compared with the conventional fertilization.

Abstract:Four kinds of activating agents (WZ,SMS,MSN,FM) were used to promote the release of calcium-magnesium phosphate (PRCMPs).Continuous water extraction and potted corn were used to study the contents of water-soluble P,fertilizer effect and phosphorus efficiency of PRCMPs.The result of continuous water extraction indicated that the content of water-soluble P of PRCMPs was higher and more stable each time compared to calcium-magnesium phosphate.The accumulative rate of release increased from 22.5% to 51.8%.The result of potted experiments showed that the biomass of potted corn from four PRCMPs were much higher than that of calcium-magnesium phosphate.Compared to calcium-magnesium phosphate,the uptaking of P of PRCMPs was improved.The contents of Mg and chlorophyll of PRCMPs were high as well.Furthermore,the residual effects of PRCMPs increased.Compared to calcium-magnesium phosphate,the contents of water-souble P,biomass of sweet corns,phosphorus uptake and soil available phosphorus were higher.The use effeciency of phosphorus also increased.The effect of the second pot experiment was better than that of the first.Among the PRCMPs,the effect of PRCMP of SMS is the best.

Abstract:A terminal software that could run on navigation PDA for recommended fertilization using ArcGIS Mobile 9.3 and abundance and deficiency indices model was developed with the support of achievement database based on GeoDatabase and mobile data service based on ArcGIS Server.Farmers can get fertilization information with the terminal software at all times and places.

Abstract:The morphological variations and discriminant methods of Channa argus (♂)，C.maculata (♀) and their hybrid were studied using traditional morphometrics and the truss network analysis.Body color of C.argus,C.maculata and their hybrid were observed.The results showed significant differences between C.argus and C.maculata from stripe at top of head,side and base of caudal fin,and the hybrid resembled C.maculata from body color.Among eleven meristic characters,the numbers of soft ray of pelvic fin were consistent in these three species,and the numbers of soft ray of pectoral fin deviated from its male parent’s species.The other meristic characters were within the range of its parent’s,the average hybrid index was calculated to be 53.45.The chi-square analysis of meristic characters showed that:there were significant differences(P<0.05) or greater significant differences(P<0.01) between C.argus and C.maculata except the number of pectoral fin rays; greater significant differences(P<0.01) between C.argus and the hybrid except the number of tail fin rays,gill rakers,lateral line scales and scales below lateral line; and significant differences(P<0.05) or greater significant differences(P<0.01) between C.maculata and the hybrid except the number of tail fin rays and gill rakers.Among meristic characters,the number of dorsal fin rays,lateral line scales and scales above lateral line can be used as preliminary indicators to determine the three species.Among ten measurable characters,the average hybrid index was calculated to be 73.90,indicating the measurable character inclined to its male parent’s species (C.argus).Cluster analysis,discriminant analysis and principal component analysis were applied to the measurable character data and the truss network data.The results indicated that the hybrid resembled C.argus from body shape,and there were greater morphological differences among C.argus,C.maculata and their hybrid.Differences between them involved the entire body.Three species could be distinguished by using the 3 formulae established with discriminant analysis which the discrimination accuracy was 100.0%.

Abstract:Eels were experimentally infected with Aeromonas hydrophila which was isolated from European eel (Anguilla anguilla) in order to evaluate the pathological changes and distribution of the bacteria in the organs at different time after the infection.First,we injected Aeromonas hydrophila into the eel muscle in five doses to determine the LD50 of this bacteria.The result showed the LD50 was 5×104 cfu/g of the eel’s body weight.Then eels were injected the bacteria in the dose of 1×107 cfu per fish into the eel muscle.We collected liver,kidney and intestine of the moribund eels at 6,24 and 48 h after the injection,and we studied the pathological symptoms and immunohistochemistry distribution of the bacteria.The result indicated that clinical signs and pathological findings were in accordance with the blood poisoning of eels.Kidney showed more pronounced lesions including necrosis in sinusoids and peritubular capillaries in the kidney compared with the other organs.Liver lesions were hyperaemia,and abiogenetic desquamation of the plasma was in intestine.The most primary distribution of the bacteria in internal organs was kidney.The challenged bacteria were also found in liver and intestine of the eels.

Abstract:Diploid (D) and tetraploid (T) loach (M.anguillicaudatus),large scale loach (P.dabryanus) (P) and their reciprocal hybrids were produced by artificial fertilization.Nine genetic crosses were made.These were (♀×♂):D×D,D×T,D×P,T×D,T×T,T×P,P×D,P×T and P×P.The relative DNA contents of the progenies were examined by flow cytometry.The results showed that D×D,D×P,P×D and P×P were diploid,D×T,T×D,P×T and T×P were triploid,T×T were tetraploid.The ratio of relative DNA contents of the three types was about 2∶3∶4.The karyotypes of the progenies were analyzed and compared by the PHA colchicine hypotonic-air drying technique.D×D has 2n=50 chromosomes,consisting of 8 metacentrics (m),6 submetacentrics (sm) and 36 telocentrics (t) chromosomes.P×P has 2n=48 chromosomes,consisting of 12 m,4 sm and 32 t.Both species have a fundamental arm number (NF) of 64.T×T has 4n=100 chromosomes,consisting of 16m,12sm,72t,NF=128.D×P and P×D are 2n=49,10m+5sm+34t,NF=64; D×T and T×D are 2n=75,12m+9sm+54t,NF=96; T×P and P×T are 2n=74,14m+8sm+52t,NF=96.By comparative analysis,the karyotypes of diploid and triploid hybrids are intermediate between the parental species,consisting of a haploid maternal set and a haploid paternal set. 

Abstract:In order to reduce the phosphorus content in the effluent and adjust the phosphorus content in artificial feed for aquaculture,a growth trial was conducted to estimate the effects of dietary phosphorous levels on growth performance,whole body composition,and serum biochemical indicators of juvenile yellow-headed catfish (Pelteobagrus fulvidraco).Three extruded diets (commercial yellow-headed catfish diet-based) were formulated to contain Ca(H2PO4)2 levels at 25,15,5 kg/t of diet (kg of Ca(H2PO4)2/t),respectively.Each diet was fed to triplicate groups of 15 fish (initial average body weight of 3.2 g) over 4 weeks.At the end of the trial,specific growth rate (SGR) and weight gain (WG) of the fish fed with the diet containing 15 kg/t was significantly higher than the other groups (P<0.05).Body crude protein content of the fish fed with the diet containing 25 kg/t had significantly increased compared with those fed the diet containing 5 kg/t(P<0.05),whereas both of them did not have significant difference between 15 kg/t treatment groups (P>0.05).Serum superoxide dismutase (T-SOD) of the fish fed with the diet containing 15 kg/t was significantly higher than the other 2 dietary treatments (P<0.05).There was no significant difference in alkaline phosphatase (AKP),calcium,and phosphorus in serum among 3 dietary treatments (P>0.05).The results suggested that phosphorus content in the current commercial feed for juvenile yellow headed catfish could be reduced from 25 to 15 kg/t,and the growth performance,body composition,and immunity of this fish would not be diminished.

Abstract:The acute toxicity of representative heavy metals (copper and cadmium),polycyclic aromatic hydrocarbons (phenanthrene and naphthalene) and pyrethroid pesticide (deltamethrin and cypermethrin) to White Cloud Mountain minnow Tanichthys albonubes were studied by using the static-test method in the laboratory.The results showed that,the 96 h-LC50 for copper and cadmium to the minnow were 0.054 and 4.610 mg/L,respectively; copper was a drastic toxicant and cadmium was a high toxicant to the minnow.And the safe concentrations of these metals were 0.005 and 0.461 mg/L,respectively.The 96 h-LC50 for phenanthrene and naphthalene to the minnow were 0.913 and 10.849 mg/L,respectively. Phenanthrene was a drastic toxicant and naphthalene was a medium toxicant to the minnow. And the safe concentrations of these PAHs are 0.091 and 1.085 mg/L,respectively.The 96 h-LC50 for deltamethrin and cypermethrin were 1.217 and 6.256 μg/L,respectively.Deltamethrin and cypermethrin were both drastic toxicants to the minnow.And the safe concentration of them are 0.122 and 0.626 μg/L,respectively.The minnow was very sensitive to these three kinds of pollutants and it could be used as a biological agent for evaluating and monitoring relative water pollution.

Abstract:This paper used mice as experimental material to establish zinc supplementation and antioxidant experiment.Biological activity of zinc chelating small peptides was investigated.The results showed that,zinc chelating small peptides can markedly improved zinc content in mice liver and serum,also can significantly reduce liver MDA content,improve the activity of T-AOC of mice.Zinc chelating small peptides has a significant effect of zinc supplementation and antioxidant,and is more effective than the same dose of zinc gluconate or zinc sulfate.

Abstract:A rape windrower was designed. The main structure and the working process of the machine were determined，including the structure design and the analysis of the key components such as the convey system, the separating system and the cutting device. A kind of convey system, which included telescopic cylinder, vertical cylinder and conveyor was designed and it could achieve the middle of the vertical placement of 1 000 mm. The cutting height can be adjusted to 200-450 mm by hydraulics equipment. The rape windrower can meet the requirement of segment harvest and was designed reasonably through experimental verification. It can extend the harvest season of rape effectively, which can not only reduce the loss rate of rape but also increase the economic efficiency.