Abstract:To identify rice mutants of floral organ and seed development,a rice mutant library of T-DNA insertional mutagenesis constructed by the national key laboratory of crop genetic improvement at Huazhong Agricultural University was screened on large scale.A large number of useful information of mutants were got.Among 9 760 stable lines of phenotype variant,177 lines showed morphology and number change of floral organ,with mutation frequency of 1.81%; 179 lines from the 3 432 lines showed abnormal seed development,the percent of mutants was 5.22%.270 lines of phenotype variant interested were obtained and 175 lines showed T-DNA insertion through PCR test,the positive rate was 64.8%,which will lay the foundation for cloning genes related with rice floral organ and seed development.One low seed-setting mutant was identified and confirmed to be cosegregated with its corresponding T-DNA flanking sequences tags,providing an important genetic resource for further studying the function of this gene.

Abstract:A constitutive gene (TIGR Locus:Loc_ Os07g34589) was obtained by combining our lab’s database with RT-PCR verification.A novel rice constitutive promoter named PSUI1 with 1 941 bp was isolated using the genomic DNA of the indica rice variety Minghui63 based on a PCR method.To determine the expression strength and pattern of the novel constitutive promoter,the cloned promoter was fused with the β-glucuronidase (GUS) reporter gene to form plant expression vector DX2181b-PSUI1,and the maize ubiquitin promoter was also fused with GUS reporter gene to form expression vector DX2181b-PUbi as the control.Both DX2181b-PSUI1 and DX2181b-PUbi were transformed into japonica rice variety Zhonghua11 by Agrobaeterium-mediated method.GUS staining showed that GUS gene was expessed in different rice tissues and organs at different growth stages of DX2181b-PSUI1 transgenic rice plants.Quantitative analysis of GUS activity showed that the strength of maize ubiquitin promoter was 2-3 times higher than that of the PSUI1 promoter,but the expression of PSUI1 promoter exhibited much more stable than that of maize Ubiquitin promoter between transgenic rice lines.

Abstract:To study the expression and function of PLT in rice,two PLT genes in rice(OsPLT7 and OsPLT11) were firstly predicted from searching the homology of Arabidopsis PLT genes protein sequences against rice database.According to the predicted promoter sequences of OsPLT7 and OsPLT11,we designed the PCR primers and amplified the consequent promoter fragments of OsPLT7 and OsPLT11 using genomic DNA of Oryza sativa L. Zhonghua 11 as templates.Finally,we cloned these fragments into the vector containing reporter genes GUS/YFP and successfully transformed these promoter-reporter fusion vectors into rice.Gus staining observation of T1 transgenic seedlings of OsPLT7 and OsPLT11 promoter-reporter fusion vectors indicated that both OsPLT7 and OsPLT11 are expressed mainly in stem cell niche and stele area in rice root,which are very similar to the expression profile of their homologous genes in Arabidopsis.These results will be useful for the functional studies of the PLT family transcription factors.

Abstract:In order to elucidate the molecular mechanism of Agrobacterium-mediated genetic transformation and improve the efficiency of Agrobacterium-mediated rice transformation by molecular biological methods,we studied the OsVIP1 protein function with the RNA interfering technology using the rice variety Zhonghua 11 as the host for genetic transformation.A rice protein sequence homologous to AtVIP1,named OsVIP1 was obtained by homologous searches.Two RNAi expression vectors,pDS1301-2-R1 and pDS1301-2-R2,were constructed with two fragments (R1 and R2) of the OsVIP1 cDNA,respectively,and used to transform rice variety Zhonghua 11 callus through Agrobacterium.The statistical results showed that formation of the transgenic resistant callus of pDS1301-2-R1 and pDS1301-2-R2 vectors was inhibited to some extent.PCR experiments indicated that R1 and R2 fragments were successfully integrated into the transgenic rice genome.Semi-quantitative RT-PCR analysis showed that the OsVIP1 gene expression was suppressed successfully in some RNAi transgenic rice plants.

Abstract:The mutant nbc(t) was a brittle culm mutant of rice found in the offspring of 93112/IRBB21.The all tissues of plant of mutant nbc(t) are brittle during the whole growth.The main agronomic characters of the brittle mutant were investigated and gene mapping of brittle trait was initially made.The results showed that the main agronomic traits of nbc(t) were as similar as the parent 9311 except that its yield was low.The rice quality of nbc(t) was also similar to 9311 except of head rice rate.The yield and rice quality of hybrid between Guangzhan 63-4S and nbc(t) were as same as that of Yangliangyou 6.Comparing with 9311,the anti-booklet strength was similar,but stretching resistance was lower.The cellulose content in culm of nbc(t) was lower than that of 9311 by 17%.The genetic analysis showed that the brittle culm trait was controlled by two pairs of dominant genes,which were mapped on two loci of chromosome 9,namely,one locus was between the SSR marker RM3700 and RM24371 with the genetic distance of 1.3 cM and 3.1 cM ; another locus was outside of the INDEL marker CL062 and CL045 with the genetic distance of 1.6 cM and 6.0 cM.

Abstract:Using ten rice varieties of different nitrogen dry matter production efficiency as materials,the effects of four nitrogen treatments on nitrogen dry matter production efficiency (NDMPE),dry matter accumulation (DMA) and nitrogen accumulation (NA) were investigated in pot experiments.The results showed that the nitrogen application increased DMA and NA,but different genotype rice had different increment.With the nitrogen application,the NA and nitrogen content (NC) and DMA increased significantly,but the DMPE obviously declined.The NA was significantly correlated with NC and DMA,the NDMPE was significantly negatively correlated with NA and NC.Significant difference of soil nitrogen apparent balance appeared among different rice varieties,and the difference become more obvious with the increase of nitrogen application.

Abstract:TPCR was used to amplify the high-frequent variable regions of AtCesA family genes,and then the PCR products were cloned into vector pGEX-4T-3,respectively.The fusion proteins composed of AtCESAs proteins and glutathione S transferase (GST-AtCESAs) in E.coli JM109 were induced by IPTG.The GST-AtCESAs were purified using GST affinity chromatography,and polyclonal antibodies were prepared.Western-blotting analysis showed that specific bands of six antibodies Anti-CESA1,Anti-CESA3,Anti-CESA6,Anti-CESA2,Anti-CESA5 and Anti-CESA8 could be obviously detected in the plasma membrane of Arabidopsis thaliana, whereas cross-reaction bands of antibodies Anti-CESA7 and Anti-CESA4 were occurred,suggesting that the two antibodies were failed for further research.Preparation of Anti-CESAs antibodies is good for future research on mechanism of cellulose synthesis.

Abstract:The combining ability and genetic parameters were estimated for four major mineral elements(Ca,Mg,Fe and Zn) in F1 progenies of 32 crossing combinations of purple Cai-tai with the incomplete diallel cross using 2 novel male sterile lines as female parents and 16 inbreed lines as male parents.The results showed that male sterile line Eru-5 and male parents D,E,F and H,which had high GCA on Ca,Mg,Fe and Zn,could be regarded as good parents.The combinations Eru-5×G,Eru-5×H and Eru-3×L had high SCA on Ca,Mg,Fe and Zn.The analysis of heritability revealed that the heredity of Ca was mainly controlled by additive effect; those of Mg and Zn were mainly controlled by additive and non-additive effects; and the heredity of Fe was mainly controlled by non-additive effects.

Abstract:The essential oil was obtained from the flowers of fresh Chimonathus praecox and dry Chimonathus praecox by simultaneous distillation extraction method and the compounds of essential oils were assayed with combined capillary gas chromatography-mass spectrometry (GC-MS).The result showed that the extracted ratio of fresh Chimonathus praecox and dry Chimonathus praecox essential oil is 0.57% and 0.71%,respectively.A total of 45 and 41 compounds were identified in fresh Chimonathus praecox and dry Chimonathus praecox,respectively, representing 92.623% and 87.078% of the total peak area.These chemical constituents mainly included caryophyllene,longifolene,β-elemene,germacrene,hinesol,α-guaiene.

Abstract:Taking the planning of regional agriculture park system in Kunming City as an example,a framework of planning regional agriculture park system was set up.The general principles of objectives for planning and development should be put forward based on the analysis of planning background and conditions of the regional agriculture parks.The layout of planning the agriculture park system consists of the layout of planning the industrial development,the spatial structure,the grade and size structure，the functional structure of the agriculture park in the region.The classification guidance plan of constructing different kinds of agriculture park,the infrastructure and key construction projects planned in the near future are important contents of planning regional agriculture park system.The innovation of institution,industry,system,mechanisms of management and operation are the important guarantee for constructing the modern agriculture parks in a region.Building a scientific evaluation system and suitable evaluation procedures and practical methods should be focused on when the construction situations of agriculture parks in a region are evaluated.

Abstract:In this report,5 261 transconjugants of Dickeya zeaewere obtained by a mini-Tn5 transposon. PCR determination revealed that 267 mutants were inserted by Tn5. Pathogenicity assay on potato indicated that they all lost or reduced the ability to cause soft rot on potato. Furthermore，the assays for pathogenicity or virulence on rice and hypersensitive reaction(HR) on tobacco showed that 71 mutants had no virulence on rice, of which 59 mutants had no HR on tobacco. In addition, 68 mutants reduced virulence in rice，The rest of mutants have no obvious difference with EC1. Moreover, 146 of 267 Tn5 mutants had no HR on tobacco, of which 90 had no or reduced the virulence on rice. The above results indicated that the mutants were produced by inserting Tn5 transposon into different genes in D.zeae chromosome. The screened mutants from our Tn5-inserted mutant library provide the basis for the further understanding of molecular mechanisms of D.zeae pathogenicity in rice.

Abstract:Apoptosis in protoplasts from homokaryon maize B37(CB37,NB37) and Mo17 (CMo17,NMo17) inbred lines treated by HMC-toxin were studied. Homokaryon maize B37 and Mo17 inbred lines were tested and the protoplasts may be stained by AO and EB. The protoplasts were divided into 4 groups. 3 groups were treated by 0,50,100,150 μg/mL of HMC-toxin as the experimental groups,and the induced time were 1,3,6 h to study the feature of apoptotic and the rate of apoptosis. The apoptotic features such as apoptotic bodies,chromatin margination or ring chromatin were found after the treatment. When treated by HMC-toxin with different concentrations,the apoptosis rates of CB37,NB37,CMo17 and NMo17 maize protoplasts were 10.80%, 4.90%, 21.00%, and 8.83%. Accordint to the above study,a conclusion can be reached as follows: apoptosis in protoplasts from homokaryon maize B37 were the same as that of Mo17; CB37 and CMo17 were more sensitive to HMC-toxin than NB37 and NMo17, respectively; when treated by the same concentration of HMC-toxin,the apoptosis rates of CB37 and CMo17 maize protoplasts were significantly higher than NB37 and NMo17 respectively; the rates of apoptosis were significantly different between C and N,and the former was significantly higher than the latter. In addition,the apoptosis rate increased when the treatment time and the concentration increased.

Abstract:All of the five cell wall-degrading enzymes,including PG,PMG,PGTE,PMTE and Cx,could be found in Brazil banana inoculated by banana wilt fusarium race 4 and Chinese banana inoculated by banana wilt fusarium race 1.PG activity was the highest.Under the condition of being cultured with seven different carbon sources,the dry weight of mycelium biomass from Foc 4 was lager than that from Foc 1,which means Foc 4 grows more quickly and is more adaptable.With the presence of citrus pectin or PGA,the two strains significantly induced PG activity.When using CMC or glucose as the sole carbon source,the PG activity of the two strains was very low.The PG activity of Foc 4 was higher than that of Foc 1 in the musa vascular tissue culture,while the PG activity of Foc 4 was lower than that of Foc 1 in the Chinese banana vascular tissue culture.With citrus pectin as a carbon source,both Foc 1 and Foc 4 can produce PG activity,but they produced lower PG activity than they did without nitrogen source.

Abstract:The resistance of 58 tomato Solanum lycopersicum materials to Meloidogyne incognita were studied by transplanting the tomatoes to disease soil in the pot. There were significant differences between 58 cultivars. The results showed that 1 cultivar was highly resistant,3 were resistant,3 were susceptible，51 were highly susceptible to Meloidogyne incognita. Degenerate oligonucleotide primers were designed according to the conserved NBS motifs among some disease-resistant genes. These primers were used for the amplification of the disease-resistant gene analog (RGA) from the genome of resistance cultivars by polymerase chain reaction (PCR), and about 500 bp-long PCR product was obtained.

Abstract:To investigate the cold adaptation of the orange wheat blossom midges,Sitodiplosis mosellana (Gehin),their round cocoons were collected and sampled from 5 different geographical districts,including Nanyang and Huixian of Henan Province,Xi’an of Shaanxi Province,Baoding of Hebei Province,and Tianjin. Results showed that the supercooling points of the round cocoons significantly decreased with the rise of the latitude. The average supercooling point was maximum in Huixian colony, which was -18.46 ℃, and minimum in Tianjin colony which was -25.34 ℃. There were large variations in Xi’an and Nanyang colony, which have the lower latitudes and lower supercooling points. Supercooling points of round cocoons had positive relationship with local extreme low average temperatures in January and February.

Abstract:A hydroponic experiment was used to study the effects of poly-γ-glutamic acid addition on the growth of Chinese cabbage and its photosynthesis characteristics,such as net photosynthesis rate(Pn),stomatic conductance(Gs),intercellular CO2 concentration(Ci),and transpiration rate(Tr).The results shows that the level of Pn,Gs,Tr and Ci in Chinese cabbage leaf treated with poly-γ-glutamate acid increases compared with control when the concentration of nutrient decreases.The application of poly-γ-glutamate acid with 90% nutrient concentration significantly increased chlorophyll a in Chinese cabbage leaves.When the nutrient concentration was 70%-100%,the above ground yield of cabbage improved compared with control,meaning that poly-γ-glutamate acid can promote transformation of light,energy capture,increase the energy efficiency through reducing the stomatal limitation,increasing the chlorophyll content and the capacity of absorption of light intensity.

Abstract:The DMRT1 gene was cloned by RT-PCR and rapid amplification of cDNA ends(RACE) methods in yellow catfish (Pelteobagrus fulvidraco).The expression of the gene was analyzed in adult tissues and different developmental stages by real-time quantitative RT-PCR.Sequence analysis revealed that the full-length of cDNA was 1 381 bp,containing 30 bp 5′-untranslated region,454 bp 3′-untranslated region \[excluding poly(A)\] and 885 bp open reading frame(ORF),which encode 295 amino acids.Alignment analysis showed that the amino acid sequences of DMRT1 gene in yellow catfish (Pelteobagrus fulvidraco) were 81%,60%,59%,64%,52%,42% and 44% identical to that from clarias lazera(Clarias gariepinus),black porgy(Acanthopagrusschlegelii),rainbow trout(Oncorhynchus mykiss),zebrafish(Danio rerio),medaka(Oryzias latipes),mouse(Mus musculus) and human(Homo sapiens),respectively.Real-time quantitative PCR analysis indicated that the DMRT1 gene expressed in the whole embryonic development stages and 51-day juveniles of postembryonic development,but the expression reached the highest level in juveniles at 31-day after hatching.During the gonad development stages,it was expressed in the male testis specifically.In the adult fish,the expression was detected only in the male testis,but not in other tissues.These results indicated that the DMRT1 gene may play an important role in the gonad formation or function maintaining of male yellow catfish (Pelteobagrus fulvidrac).

Abstract:The first-strand cDNA sequence encoding pancreatic trypsinogen (TRY) gene was isolated from yellow catfish (Pelteobagrus fulvidraco) larvae (1-30 dph,day post-hatching) by RT-PCR,and using real-time PCR to detected and quantitated the trypsinogen gene expression.The results showed that the onset of trypsinogen gene expression in P.fulvidraco larvae was 1 dph and the trypsinogen gene expression increased significantly from 1 to 6 dph,increased fluctuantly from 6 to 23 dph and rose sharply from 23 to 30 dph.The experimental results demonstrated that the detection and quantitation of trypsinogen gene expression in P.fulvidraco larvae would accurately reflect the process that digestive functioning developed in intestinal tract.And we suggested that weaning time of P.fulvidraco larvae would be much better after 23 dph.

Abstract:In this paper,the inter-simple sequence repeat (ISSR) technique were applied to analyze the genetic diversity among 5 color patterns of Oujiang color carp,designated as whole red,whole white,red color with small black spots,white color with large black patches,red color with large black patches,fourteen specific and stable primers were screened from 60 primers and a total of 125 loci were generated,of which 85 loci (68%) were polymorphic.The results indicated that effective number of alleles (Ne),expected heterozygosity(h) and Shannon’s information index(I) were 1.479 3,0.273 9 and 0.401 0,respectively.Genetic similarity (GS) and genetic distances (GD) were calculated by using Nei’s matching coefficient,the average GD among 5 color patterns of Oujiang color common carp is 0.342 4.When NJ and UPGMA cluster analysis were applied,Oujiang color carp were divided into two groups,named whole white,white color with large black patches and whole red in one group,red color with small black spots and red color with large black patches in the other group.Subsequently,the 3 polymorphic bands amplified from the primer 848 were employed to construct the DNA fingerprints of the 5 color patterns of Oujiang color carp.Among the constructed DNA fingerprints,each color pattern of Oujiang color carp had its unique fingerprinting pattern and could be easily distinguished from each other.Taken together,the results in this study show that the ISSR marker is a useful technique for identification of Oujiang color carp,offering the more correct and quicker results.

Abstract:Five experimental diets for gibel carp,Carassius auratus gibelio,were formulated to contain various percentages of common rapeseed meal (CRM) or double low rapeseed meal (DLRM) as a partial replacement for soybean meal.RM0(with 0% CRM and DLRM) was formulated as the control group.DLRM50 (CRM50),DLRM100(CRM100) were formulated to isonitrogenously replace 50% and 100% of soybean meal protein in RM0 with DLRM (CRM),respectively.The non-specific immune parameters including phagocytic activity (PP,PI) of blood leucocytes and head kidney phagocytes,serum lysozyme activity,serum complement concentrations of C3 and C4 were evaluated at every 14-day intervals.During the initial 28-day period,only PP of blood leucocytes in CRM100 group was significantly lower than that in RM0 treatment(P<0.05).From the 42th day on,both PP and PI of blood leucocytes and head kidney phagocytes in DLRM100 and CRM100 treatments were all significantly lower than those in RM0 treatment (P<0.05).On the 56th day,serum lysozyme activity in CRM100 and DLRM100 groups was significantly lower than that in RM0 group(P<0.05).From the 42th day,serum complement concentrations of C3 or C4 in DLRM100 and CRM100 treatments were significantly lower than those in RM0 group(P<0.05).The immune parameters in DLRM50 and CRM50 treatments were not significantly changed compared with those in RM0 group (P>0.05).However those in CRM50 group gradually reduced with time.We concluded that the effects on non-specific immune function might cause by antinutritional factors such as glucosinolates,sinapin,phytic,tannin,crude fibre in the rapeseed meal.

Abstract:The effects of the dietary complex Chinese herbs on non-specific immune function and disease resistance in yellow catfish (Pelteobagrus fulvidraco R.) were studied.The yellow catfish were divided into 6 groups and were fed with feed supplemented with different ratios of complex Chinese herbs，0 mg/kg（control group,200 mg/kg（groupⅠ）,400 mg/kg （group Ⅱ,600 mg/kg（group Ⅲ）,800 mg/kg（group Ⅳ）and 1 000 mg/kg （group Ⅴ),respectively.Thirty days later,the yellow catfish were blooded,and the leucocytes phagocytic activity,serum lysozyme activity,complement activity and disease resistance were monitored.The results showed that the phagocytic activity of the leucocytes were significantly higher than the control group (t-test,P<0.05).The lysozyme activity of the serum in group Ⅲ,Ⅳ and Ⅴ were significantly higher than the control group (t-test,P<0.05)，but there were no significant differences between group Ⅰ,Ⅱ and the control group (t-test,P>0.05).The complement activity of the serum in group Ⅱ,Ⅲ,Ⅳ and Ⅴ were significantly higher than the control group (t-test,P<0.05),while there were no significant differences between group Ⅰ and control group (t-test,P>0.05).The results showed that the best supplementation ratio of the complex Chinese herbs for the yellow catfish was about 400-800 mg/kg.

Abstract:The testis structure,development regularity and spermiogenesis of sexually mature Coilia nasus were studied by using paraffin sections and H.E.staining method.The results indicated that,the male gonad of Coilia nasus was composed of testis (sperm production),seminal vesicle and spermatic duct.The spermary had typical lobular structures,which is composed of seminiferous lobula,follicle,interstitial substance,lobular cavity and efferent duct.The seminiferous lobula was composed of spermatogenic cells of varied stages and supporting cells,and spermatogenic cells of varied stages and supporting cells formed the follicle.The mature sperm was released from the follicle into the lobular cavity,then entered into efferent duct via transient pathway formed between lobes,and then was sent into the seminal receptacle.The seminal receptacle was in reticulated structure,and the inner wall was composed of excreting cells.The sperms develop through 5 stages,including androgone,first spermatocyte,secondary spermatocyte,spermatoblast and mature sperms.Six stages were classified according to the appearance,color,volume and vascular distribution of the spermary.

Abstract:In order to analyze the variation of loading in the process of shear and whether different loading rates have an impact on shear strength and shear energy or not, four different species of wheat stalks in mature stage and dried under natural conditions were selected, which were Yibi No.1, Aikang 58, Zhoumai 22 and Yumai No.7, and the stems of their second, third and fourth internodes were chosen for study. The middle of each internode was tested for cross striations shear on the TMS PRO type Texure Analyzer produced by the U.S.FTC company. Through the shear testing, the biomechanical properties of the maximum loading, shear strength and shear energy were measured for various kinds of wheat stalks on different internodes with different speeding rates. The study result indicates that the variation of loading in the process of shear was that the loading increased at the beginning, then reduced because of its structural features, then added rapidly until to rupture. Meanwhile, the hardness, shear strength and shear energy on different internodes of the tested wheat stalks were respectively within the ranges of 37.3~191.0 N, 4.2~9.8 MPa,and 43.53~432.23 mJ. As for the shear strength and shear energy on different internodes of the same species of wheat stalks, those on the second internode is the largest, the fourth is the least. At last the significance test for shear strength and shear energy was run by applying SPSS, the result shows that different speeding rates have no significant impact on shear strength and shear power.

Abstract:The characteristics and principles of seed treatment methods commonly used in crop production such as physical,chemical,biotic pesticides,seed priming,seed coating and seed pelleting were summerized.The biological effects of seed treatment were mainly summarized and analyzed.Meanwhile,the trend of crops seed treatment was prospected.