Abstract:The sequence between translation initiation site of a bidirectional gene pair(TIGR Locus:LOC_Os09g39540 and LOC_Os09g39550) has been predicted as a bidirectional promoter. This promoter was acquired from the genomic DNA of Minghui 63 by PCR and named as BDP1 and inserted into the promoter function analysis vector pDX2181 in forward and reverse directions (named BDP11 and BDP12). BDP1 was identified as an endospermspecific bidirectional promoter in transgenic plants with GUS histochemical staining. The expression efficiency of the two directions were low. The promoter fragment was extended 123 bp downstream from the end nearer the transcription start site. The new promoter named as BDP2 was inserted into the promoter function analysis vector pDX2181 in forward and reverse directions (named as BDP21 and BDP22). The expression pattern of the BDP2 was the same as BDP1. The expression efficiency of BDP21 and BDP22 were higher than that of BDP1.

Abstract:The effects of basal medium and explants on the primary culture of dormant buds of date plum (Diospyros lotus Linn.) were studied.The effects of inoculation methods,TDZ concentrations,combinations of plant growth regulators and induction period under dark condition on callus formation,adventitious bud regeneration and root regeneration were investigated.The results showed that DKW was the best basal medium for the primary culture of dormant buds,as revealed by better growth of the buds.Genotypes had no impacts on primary culture,but influenced subculture obviously.Inoculation of the segments with the adaxial surfaces contacting medium on MS+ZT 0.1 mg/L+TDZ 2.0 mg/L under darkness for 20 d led to 100% formation of callus,but the percentage was only 81.4% on (1/2N)MS+2.200 mg/L TDZ.Inoculation of leaf segments,adaxial surfaces contacting medium,on MS medium containing ZT 1.0 mg/L+TDZ 2.0 mg/L,dark treatment for 0 d,resulted in the 34.4% of shoot regeneration,while average shoots numbers per explant were 2.1±0.3.Inoculated the plantlets on 1/2MS(1/2N)+IBA 1.0 mg/L for 5 d,then transplanted them on 1/2MS(1/2N)+1 g/L activated carbon for 20 d.Rooting of date plum reached 58.14% and the average number of adventitious roots was 4.

Abstract:The effects of basal medium and explants on the primary culture of dormant buds of date plum (Diospyros lotus Linn.) were studied.The effects of inoculation methods,TDZ concentrations,combinations of plant growth regulators and induction period under dark condition on callus formation,adventitious bud regeneration and root regeneration were investigated.The results showed that DKW was the best basal medium for the primary culture of dormant buds,as revealed by better growth of the buds.Genotypes had no impacts on primary culture,but influenced subculture obviously.Inoculation of the segments with the adaxial surfaces contacting medium on MS+ZT 0.1 mg/L+TDZ 2.0 mg/L under darkness for 20 d led to 100% formation of callus,but the percentage was only 81.4% on (1/2N)MS+2.200 mg/L TDZ.Inoculation of leaf segments,adaxial surfaces contacting medium,on MS medium containing ZT 1.0 mg/L+TDZ 2.0 mg/L,dark treatment for 0 d,resulted in the 34.4% of shoot regeneration,while average shoots numbers per explant were 2.1±0.3.Inoculated the plantlets on 1/2MS(1/2N)+IBA 1.0 mg/L for 5 d,then transplanted them on 1/2MS(1/2N)+1 g/L activated carbon for 20 d.Rooting of date plum reached 58.14% and the average number of adventitious roots was 4.

Abstract:To study the genetic diversity of pollens of sand pear germplasm,pollens quantity and pollens germination of 455 sand pear germplasm were investigated in vitro .The results showed that there were significant variations on the quantity of the total pollens and germination rate among the sand pear germplasm,ranging from 0 to 41 875 grains per anther and from 0 to 96.11%,respectively．The average coefficient variation and genetic diversity index was 66.37 and 1.95 of pollen characteristics.The average variation coefficient of different sand pear germplasm was in the order of China improved lines (67.12)>landraces (64.58)>foreign breeding cultivars (61.08).The average genetic diversity index was in the order of landraces (1.95)>China improved lines (1.89)>foreign breeding cultivars (1.81).The average coefficient variation of landraces in different provinces (regions) ranged from 53.51 to 95.90,with the highest in Guangxi and the lowest in Fujian.The genetic diversity index was between 1.55 and 1.88, with the highest in Hubei and the lowest in Jiangxi.

Abstract:To study the effects of white clover culture on soil fertility and fruit quality of Cara Cara naval orange (Citrus sinensis Osbeck),soil available mineral elements content of orchard were measured 5 years after sod culture (in 2015).The fruit qualities of Cara Cara naval orange were measured 45 years after sod culture (in 2014 and 2015).The results showed that available P (phosphorus) and Fe (iron) content were significantly up regulated in white clover culture treated soil compared with the control.The white clover culture significantly increased fruit weight,fruit shape index and edible rate compared with the control.Fruit weight in 2014 and 2015 was increased by 15.71% and 28.98%.Fruit peel thickness thinning in 2014 and 2015 was increased by 0.28 mm and 0.18 mm.Fruit shape index was increased by 0.04.Fruit edible rate in 2014 and 2015 was increased by 2.3% and 2.1%.The total soluble content,total acid and vitamin C of fruit were not significantly affected under the white clover culture.The white clover culture increased soil fertility of orchard and improved fruit quality of Cara Cara naval orange.The white clover culture has the potential to be popularized on Cara Cara naval orange orchard.

Abstract:The male sterile line BY used as the female parent was crossed with six inbred lines of Tagetes patula including K4,K8,K13,K15,K17,K30 andtwo Tagetes erecta including 9904 and 9906 in two consecutive years to study the application potential of the male sterile plant. Seed setting rate,the main qualitative and quantitative traits of the eight crosses were investigated. The results showed that the seed setting rates and germination rates of intra specific hybrids were relatively high in two years. The seed setting rate of hybrids between T. patula and T. erecta was extremely low due to the cross incompatibility. When T. patula+E26 was used as male parent,flower type of hybrids was determined by the male parent with double petal form and flower color was partial to red and orange. When T. erecta was used as male parent,flower type,petal form and flower color were similar to the female parent. All were cylindrical,lack of appreciation. Comparing the mean value of main traits,BY×K17 was an excellent combination with low plant height,small leaf blade,large flowers,long viewing period,high seed setting rate and germination rate. Both BY×K30 and BY×K8 had a good view because of large flowers and long viewing period. The results of over maleparent heterosis showed that when T. patula was used as male parent,the heterosis of other traits were positive except that of flower diameter (-0.64%). when T. erecta was used as male parent,the heterosis of other traits was based on characteristic of male parent except that of diameter of inflorescence. 

Abstract:The inhibition of cell proliferation,cell morphological change and mitochondrial membrane potential change of ethanol extracts of 20 Thailand plants on Spodoptera litura (Sl-1) cell were investigated by MTT test,inverted phase contrast microscope and flow cytometry,respectively.The results showed that the Sl1 cell proliferation inhibition effect of 5 ethanol extracts including Thailand plants (Zingiber officinale Roscoe, Alpinia galangl (L.) Willd.,Azadirachta indica A.Juss var.siamensis Valeton,Curcuma longa Dennst and Citrus hystrix DC) was better than that of the control rotenone,and the inhibition was dosedependent. After 24 h of the treatment,cells showed obvious morphological changes,including adhesion capacity reduction,vacuole and debris appearance.After 48 h of the treatment,more and more vacuoles and debris were present obviously in the media.Moreover,these five ethanol extracts could induce cell mitochondrial membrane potential change,among which A. indica A. Juss-var. siamensis Valeton and C. hystrix DC were more effective.In conclusion,Z. officinale Roscoe,A. galangl (L.) Willd.,A. indica A. Juss var. siamensis Valeton,C. longa Dennst and C. hystrix DC possessed cytotoxicity on Sl-1 cells and deserved further researches.

Abstract:To determine the types,signal peptide,and conserved motif of the type Ⅵ secretion proteins (Tss) in Xanthomonas campestris pv. raphani 756C ( Xcr),and to provide an important theoretical guidance for clarifying the Tss of the other Xanthomonas pathogen,Tss sequence of Xcr from the protein databases was searched using keywords,and the conserved domain was searched using SMART online.The signal peptide,transmembrane domain structure,the tertiary structure and conserved motif were analyzed.Meanwhile,the genetic relationships was analyzed using comparative Tss in Xcr and homologous sequences of other species.There are three Tss in Xcr ,namely TssA,TssB,TssC,which have more than 50% helical secondary structure and only TssC contains the typical signal peptide sequence.All the 3 Tss positioned in the plasma membrane,and each has 3 conserved motifs.Based on the genetic analysis of Tss,it is suggested that Xcc and Xca are closely related to Xcr.

Abstract:Streptomyces carneohygroscopicus SH 121 isolated during selecting biocontrol agent of citrus diseases exhibits strong inhibition towards Gram positive bacteria and many pathogenic fungi in plant.Establishment of a highly efficient genetic manipulation system is critically required for functionally analysing genes and genetic engineering in Streptomyces.In this study,the effects of heat treatment,pregermination time,donor hosts,ratio of donor and recipient and MgCl2 concentration in medium on conjugation efficiency were studied by single factor experiments.Under heat treatment at 50℃ for 10 min,pregermination at 37℃ for 1 h,using Escherichia coli DCDA/pUZ8002 as donor host,with the ratio of donor to recipient being 100 to 1,supplementing MISP4 medium with 40 mmol/L MgCl2,a highly efficient intergeneric conjugation method for SH121 was developed with highest efficiency of 1.03×10-3 conjugants/ recipient.The integrative plasmid pSET152 and the plasmid pYZ09 for gene knockout was successfully transferred intoSH121 with the method newly established. It is indicated that this genetic manipulation system could be used to discover bioactive natural products and study their biosynthetic gene clusters in SH121.

Abstract:Based on analyzing the transcriptome of Mesorhizobium huakuii 7653R, MCHK_8182 was identified to be upregulated significantly during the stage of symbiotic nitrogen fixation.SOEPCR primers were designed and an MCHK_8182 replacement vector was constructed.Through three parental matting,the replacement vector was introduced into M.huakuii 7653R.The transconjugants were screened under a pressure of sucrose culture.The MCHK_8182 replacement mutant was finally obtained and its correctness was confirmed by PCR and sequencing.The result of the pot experiment showed that the phenotype of host plants induced by MCHK_8182 mut had a remarkably reduced number of root nodules compared with wildtype 7653R,but nodules nitrogenase activity remained unchanged.

Abstract:Optimizing fermentation conditions of endocellulase production by a psychrotrophic bacteria using response surface method

Abstract:The fraction of absorbed photosynthetically active radiation (FPAR) is an important parameter to estimate the net primary productivity (NPP) of terrestrial ecosystem by remote sensing.However,when the leaf area index (LAI) and other parameters increase,FPAR becomes less sensitive to NPP at the same time.In order to analyze a more accurate linear relation between NPP and FPAR,some scholars point out a concept in allusion to the absorbed photosynthetically radiation of chlorophyllFPAR absorbed by photosynthetically active vegetation (FPARgreen)which is more accurate and helpful for monitoring and analyzing crops compared with FPAR.Nowadays FPARgreen is extracted by vegetation index inversion,and its final inversion precision is affected by the vegetation index selected.The PROSPECT+SAIL model was used to calculate the true FPARgreen.Various common vegetation indices were stimulated to make linear fitting with the linear fitting precision of each vegetation indices.Some parameters including leaf inclination angle and soil reflectance were modulated to screen the best vegetation index.

Abstract:Blood parameters are important phenotypic metrics for individual antivirus capacity.To investigate the effects of polyinosinic:polycytidylic acid (Poly I:C) stimulation on porcine blood parameters,we made correlation and principal component analysis on porcine blood parameters before and after Poly I:C stimulation.A total of 392 experimental pigs were selected from a DurocErhualian F2 population.The blood samples from each pig were collected by jugular venipuncture on day 33 (the control group) and at 4 h after Poly I:C stimulation on day 35 (the treatment group).The results showed that NE%,LY and LY% were significantly correlated,and HCT% and RBC were significantly different before and after the stimulation.The PLT and PCT,which are the decisive indices of the first principal component in the blood platelet,were not significantly different before and after the stimulation.

Abstract:In the present study,differences of spatial learning and memory ability between male and female bitterling, Rhodeus sinensis,were investigated using Tmaze.A faster decline in mean latency and higher percentage of correction were detected in the trained female bitterling.The times of training trials reaching the criterion was also lower in female than that in male.The results showed that,both male and female bitterlings were able to learn a spatial task after being trained.However,there were significant gender differences in reproduction season,with the capability of spatial cognition in female excelled that in male.

Abstract:Effects of 4 common preservation methods (5% formalin,10% formalin,95% alcohol and -20℃ frozen) on spore morphology of Myxobolus wulii were investigated in the present study.Compared with fresh spores,only the spores preserved in 5% formalin for 7 d showed no significant differences in measurements (P<0.05),which indicated that 5% formalin is a suitable fixative for M.wulii spores preservation for a short period of time (less than 7 d).The change rates of measurement indexes of preserved spores were different,indicating that the present preservation methods made M.wulii spores deform irregularly.For spores preserved by different preservation methods in the same time period,95% alcohol yielded the maximum contraction,following by -20℃ frozen,10% formalin and 5% formalin.The total change rates of spores,except for that preserved in 10% formalin,decreased with the extension of preservation time,indicating that the preserving spores doesn’t always shrink continuously.

Abstract:To characterize the variant porcine epidemic diarrhea virus (PEDV) and lay the foundation for prevention and control of porcine epidemic diarrhea (PED),a variant PEDV strainCH/YNKM8/2013 was isolated in 2013 in China.The strain was identified by cytopathic effects (CPEs),RTPCR,IFA,and transmission electron microscope (TEM).The genome sequence of it was sequenced by segmented amplification method and was deposited at GenBank under the accession number of KF761675.1.The Nterminal of S gene of CH/YNKM8/2013 was 9 bp longer than that of CV777 strain,including 15 bp insertions and 6 bp deletions,which suggested that CH/YNKM8/2013 was a variant PEDV strain.The CH/YNKM8/2013 had high sequence similarity of 99.6% with the Chinese strain LC (isolated in 2011 in Guangdong province),of 99.0% with the Korea strain KNU1305 and the USA strain USA/Minnesota84/2013 and a lower sequence similarity of 96.7% with the vaccine strainCV777,respectively.The phylogenetic analysis showed that CH/YNKM8/2013 was clustered into the same group with Chinese,South Korea and USA variant PEDV strains,while it was clustered into different branch with former Chinese classical PEDV strains and the vaccine strainCV777.

Abstract:The effects of pulsed microwave on physicochemical,rheological properties and sensory quality of rice were studied using ordinary continuous microwave as control.The results showed that the temperature,broken rice rate,percent cracked rice and blue value of microwavetreated rice were higher than that of untreated rice,except for the moisture content.The percent cracked rice after pulsed microwave treament (10%) was significantly lower than that of ordinary continuous microwave (24%),close to untreated rice (8.5%),indicating that the effect of pulsed microwave was better than that of ordinary continuous microwave.In steady shear mode,all rice pastes were typical nonNewtonian fluid with pseudoplastic behavior.Compared with untreated rice paste,the viscosity of rice paste treated with microwave decreased and the mobility increased,close to a Newtonian fluid.In dynamic oscillation mode,the pasting temperature of both untreated and microwavetreated rice were at ~68℃.The storage modulus (G′) and loss modulus (G″) of microwavetreated rice paste was higher than that of the untreated rice paste,indicating that the three dimensional gel network in the rice paste could be enhanced by microwave.There was no significant changes in the sensory qualities of rice treated by pulsed microwave.The shape score of cooked rice treated by pulsed microwave was significantly higher than that of ordinary continuous microwave.The pulsed microwave will have a potential application in the processing and storage of food.

Abstract:The browning related enzymes in mango skin were investigated.Crude proteins were extracted from mango juicing with centrifugation,filtration and concentration methods and purified by column chromatography.Two isoenzymes,P1 and P2,were purified.Using catechol as substrate,the activity of the enzyme was determined by spectrophotometry at 420 nm.The results showed that the optimal temperature,pH value,substrate for mango P1 was 55℃,7.0,and catechol,respectively.The molecular weight of P1 was 100 ku.The activity of P2 was very low.Results of circular dichroism showed that the content of αhelix and βfold of the two enzyme was 32.1% and 19.2%,43.4% and 80.8%,respectively.Result of fluorescence excitation spectra showed that the maximum emission wavelength of P1 and P2 was at 358.4 nm and 356 nm with the fluorescence intensity of 207.9 and 164.2.The particle size distribution of the two enzymes was more concentrated at 11.696 nm and 8.721 nm,reaching 29.686% and 25.369%.

Abstract:In mountainous orchard of southern China,currently the selfloading devices with the Ltype pull arm were mostly put into use in leveling roads with large volume which is not suitable for the mountainous region. In order to protect the fruits and improve the transport efficiency,a selfloading device with the Ltype pull arm in mountainous orchard transporter was designed,and simulations were conducted. The stress of the Ltype pull arm was analyzed with the traditional graphical method to calculate the formula of main mechanical stress in the course of two campaigns,and the critical angle was 38 °. Getting the relation between the main part of power and the rotation angle by ADAMS. Results indicated that there were four cases regarded as dangerous conditions: slope of -20° and rotation angle of pull arm of 0°,slope of 20° and rotation angle of pull arm of 38°,slope of -20° and rotation angle of pull arm of 134°,slope of -20° and rotation angle of pull arm of 38°. Results of ANSYS finite element analysis declared that most of stress of the pull arm was in the range of 0.0768.316 MPa. The maximum stress was 307.144 MPa,less than 470 MPa of the steel Q345’s minimum tensile strength. This design has met the demand to complete the requirements of loading and unloading on the working conditions that the slope of 15° and the load of 150 kg.

Abstract:A deepshallow rotary tillage device was designed to reduce the operation power consumption of direct rapeseed seeder.Installed in a way of symmetric helix arrangement with multiple section,the device blades divided seed bed into seed bed strip rotating deeply and stubblechopping strip rotating shallowly and chopping stubble by cross arrange.The main design parameters of seed bed strip,stubblechopping strip and the blade arrangement were decided.The result of field test showed that the power consumption of rotary part was 72.53%82.20% of whole power consumption of direct rapeseed seeder under certain condition of field test.Compared with the original rotary tillage device,the operation power consumption of deepshallow rotary tillage device decreased 14.13%18.04%.The value of working evaluation indexvegetation buried rate,soil breaking rate and smooth field surface decreased,meeting the standard requirement of rotary tillage device.The smooth seed bed and steady ditch width were obtained with analyzing the seed bed outline of field test operated by deepshallow rotary tillage device.It is indicated that the deepshallow rotary tillage device can reduce the operation power consumption of direct rapeseed seeder.It will provide reference for further optimization.

Abstract:To solve problems including low leveling precision and poor controllability etc.of paddy field rotary cultivator after plowing caused by tilting and vibration while farming,a set of leveling system consistent with paddy field rotary cultivator was designed on the basis of tilt angle sensor control technology.It realized leveling regulation function by combining hydraulic control with the controller control.The results of field trials showed that the working performance of field rotary cultivator leveling system based on tilt angle sensor technology was practical with its automatic leveling regulation function.The flatness after plowing Sd was 2.20 cm，with elevation difference distribution β of 81.82%. Compared with field rotary cultivator,the field surface roughness was 34.3% and the elevation difference distribution increased 19.4%，indicating that it can basically meet the agricultural requirement of rice planting.

Abstract:The sizes and the uniform performances of peanut seeds are the basis of seed cleaning and grading,and the core element of breeding peanut.Three axis dimensions,the average diameter and weight per 100 seeds were selected as indicators.The differences,distribution and interrelation of the three axis dimensions of the seeds,the uniform performance and the relationship between seed sizes and the weights were analyzed with statistics.The results showed that three axis dimensions obeyed the normal distribution with significant differences among different varieties of peanuts.There were linear correlations between three axis dimensions.Differences between seeds of peanut varieties were homogeneity.The geometric mean diameter of peanut seeds,arithmetic mean diameter and the trends of weight changes were basically same.The sizes and the uniform performances were significantly different among peanuts varieties.Many factors should be comprehensively taken into account on cleaning and grading of peanuts seed.