【Objective】 The objective of this experiment is to investigate the expression profiling of rice calcineurin B-like (CBL) proteins and the mechanism of stress-tolerance.【Method】The expression patterns of CBL proteins at seedling stage of super hybrid rice parental line 9311 under cold, hot, drought, submerge and salt stresses were surveyed by Western blotting (WB) via antibody-based proteomics strategy. 【Result】The results indicated that the expressions of CBL1, CBL5 and CBL6 were down-regulated under cold stress, the expressions of CBL1 and CBL2 were down-regulated under hot stress, the expressions of CBL1 and CBL5 were down-regulated while CBL6, CBL8 and CBL10 were up-regulated under drought stress. The expressions of CBL1, CBL5, CBL6, CBL8 and CBL10 were up-regulated and CBL2 was down-regulated under submerge stress. 【Conclusion】 Six abiotic stress-related rice CBL proteins were identified.

【Objective】 The objective of this study was to isolate HvGW2 gene from barley BAC library rapidly. 【Method】 Three-dimensional (3D) BAC pools were constructed and screened by using barley GW2 gene-specific PCR primers. Positive BAC clones were screened out and further sequenced, the obtained HvGW2 gene was used for homologous evolutionary analysis with those of other grasses. 【Result】 One hundred and sixty BAC DNA pools were prepared by 3D pooling strategy from barley BAC library with only 0.18% of empty clones. PCR-based screening demonstrated that only one of the three positive BAC clones (196M02) contained the complete coding sequence of HvGW2. Sequence analysis showed that barley GW2 gene contains eight exons and seven introns, which revealed that it has similar gene structure and conserved functional domain with other plant species. Furthermore, HvGW2 has the highest homologous with wheat GW2 protein. Moreover, the sixth intron of HvGW2 gene contains two different retrotransposon insertions, which resulted in a 11.5 kb length increase of barley GW2 gene than the other crops.【Conclusion】 By constructing three-dimensional BAC pools, and combining with PCR-based screening, positive BAC clones which contain target gene can be isolated quickly. Not only the quality of the barley BAC library was evaluated, but also an auxiliary platform for barley gene cloning was established. It was speculated that the retrotransposon insertions in gene intron might be the key variation which affected the gene transcription levels of HvGW2.

【Objective】A method for fast, non-destructive and accurately monitoring leaf water content (LWC) of wheat (Triticum aestivum L.) was improved with hyperspectra technology in this paper. 【Method】The canopy leaf spectral reflectance in main growing seasons of wheat was collected under the condition of water stress experiment. Using the frist-order derivative spectra, 16 new hyperspectral indices were developed to quantify the wheat’s leaf water content (LWC). These indices were then compared with the commonly used hyperspectral indices including NDII, WBI and NDWI to screen out the spectral index which was sensitive to LWC for modeling. Using the inversion model, the OMIS image was calculated one pixel by one pixel, and the remote sensing mapping for LWC of wheat was accomplished. 【Result】The accuracy of inversion model (logarithmic function) which built by index FD730-955 was higher than that by the hyperspectral indices commonly used, as indicated by a calibration determination coefficient (C-R2) of 0.749 and a validation determination coefficient (V-R2) of 0.742. The eatimation values of 32 samples in prediction set were close to the measured values, and R2 and RMSE of regression fitted model between two dataset were 0.763 and 0.024, respectivly. Furthere more, the prediction accuracy of FD730-955 was least sensitive to the change of LWC and LAI among all of the hyperspectra indices and therefore least affected by the range of sample values and canopy density when used to estimate the LWC of wheat. The R2 and RMSE of the fitting model for the inversion and measured values were 0.635 and 0.027, respectively, and indicated the similarity between the inversion and measured value was high. 【Conclusion】 It is possible to eatimate LWC of wheat by derivative hyperspectra with a high accuracy, and FD730-955 is an optimal index for modeling.

【Objective】 Understanding the effects of water stress during grain filling on physicochemical properties of waxy maize flour.【Method】A negative pressure water supplying and controlling pot device was used to control different soil moisture contents (SMC) by setting the water supply tension of the device at different values. The effects of water stress (relative SMC was 90%, 75%, and 60%, indicated the water supply was excessive, moderate, and deficient, respectively) during grain filling on physicochemical properties of waxy maize flour were studied using Suyunuo5 and Yunuo7 as test materials.【Result】Among different pasting characteristics, trough viscosity, final viscosity, setback and pasting temperature were not affected by water status, while the lower peak viscosity and breakdown were presented when SMC was deficient. The results of differential scanning calorimetry observed that enthalpy of gelatinization and conclusion temperature were similar among different water treatments; onset and peak temperature was higher at deficient water supply treatments. Compared with moderate SMC, both excessive and deficient SMC increased the percentage of retrogradation. The average granule diameter and percentage of starch granule size that diameter ＞17 μm were the lowest when SMC was moderate for Suyunuo5, while it was the highest for Yunuo7. Λmax was the lowest when SMC was moderate and it was the highest when SMC was deficient for both varieties. Iodine binding capacity and crystallinity were stable among different SMCs for Suyunuo5, while the higher iodine binding capacity and lower crystallinity were presented when SMC was deficient for Yunuo7. 【Conclusion】The lowest crystallinity and highest iodine binding capacity at deficient SMC treatments resulted in the lowest peak viscosity and breakdown, and highest onset temperature, peak temperature and percentage of retrogradation. The waxy maize flour presented better processing quality (higher peak viscosity and breakdown, low percentage of retrogradation) at moderate SMC treatments.

【Objective】The objective of this study is to realize the digitization and visualization of tobacco. 【Method】 In this paper, the template technique was used for three-dimensional reconstruction of the tobacco plant and visualization. The measured data of tobacco organic were firstly obtained using 3D digital instrument and image processing; then the simulation for tobacco leaves, flowers, fruit and stems were realized accordingly to different parametric descriptions based on morphological characteristics, and saved them as template files. At last, the complete model was constructed with graphical transformation operations on them. 【Result】 Using this method can facilitate the construction of the major periods of tobacco with 3D model.【Conclusion】 The result shows that the tobacco model rebuilt by this method has more reality, which offers a technique support for its iteraction design and visualized simulation.

【Objective】 The objective of this study is to investigate the roles of NSvc4 and CP proteins of Rice stripe virus (RSV) in pathogenicity to reveal the mechanisms underlying RSV pathogenesis. 【Method】 The subcellular localization patterns of six RSV proteins were studied by expressing them in epidermal cells of Nicotiana benthamiana through Agrobacterium inoculation. The interaction between NSvc4 and five other proteins of RSV was investigated using Bimolecular fluorescence complementation (BiFC), and the interaction between NSvc4 and CP was further confirmed by yeast two-hybrid experiments. The possible involvement of NSvc4 and CP in pathogenicity was exploited by expressing them individually or together in N. benthamiana employing the vector Potato virus X (PVX). 【Result】 NSvc4 interacted with CP and they formed protein complexes in chloroplasts of N. benthamiana when co-expressed, both NSvc4 and CP could induce leaf mosaic symptoms when they were expressed using PVX. However, leaf mosaic symptoms caused by CP was recovered in systemic leaves of PVX inoculated plants. When expressed together or as a fused protein using PVX, NSvc4 and CP induced severe symptoms in both inoculated and systemic leaves of N. benthamiana. Real-time PCR experiments showed that NSvc4 and CP had no effects on the accumulation of PVX, and the severity of the symptoms induced by the recombinant virus correlated with the expression levels of NSvc4 and CP. 【Conclusion】 Both NSvc4 and CP have pathogenicity properties and they might have synergistical function in RSV pathogenesis.

【Objective】The objective of this study is to identify the most tolerant stage of azuki bean beetle, Callosobruchus chinensis L. (Coleoptera: Bruchidae) to low and high temperatures, and obtain different temperature-exposure time to kill the various stages of the insect in storage. And to provide a theoretic foundation for popularizing and applying low and high temperatures for the control of C. chinensis.【Method】Four development stages were exposed for different times at -5 and 40℃ to get the most tolerant stage of C. chinensis for low and high temperatures. Based on the above experimental results, the most cold-tolerant stage was used for consequent bioassays at -5, -10 and - 20℃, and the heat-tolerant stage at 40, 45 and 50℃. 【Result】 Pupa and larva were more cold tolerant stages, LT50 at -5℃ for eggs, larvae, pupae and adults were 12.57, 24.93, 30.54 and 15.76 h, respectively. Pupa was relatively more heat tolerant than other three stages, the LT50 at 40℃ for eggs, larvae, pupae and adults were 4.29, 17.76, 22.33 and 14.50 h, respectively. The LT50 for pupae were 30.54, 6.50 and 0.96 h at -5, -10 and -20℃, respectively. The LT99 were 189.70, 33.81 and 2.90 h, respectively. The LT50 of pupae at 40, 45, and 50℃ were 22.33, 3.64 and 0.85 h, respectively. The LT99 were 169.43, 17.77 and 3.71 h, respectively. 【Conclusion】Both cold tolerance and heat tolerance of pupa are higher than other three life stages, and different temperature-exposure times to kill the pupa stage of the insect are obtained. It is feasible to utilize low and high temperatures to control C. chinensis in storage.

【Objective】The objective of this study is to research the preparation prescription of KillerRed protein liposome (L-KRP) and its physicochemical properties and bioactivity for optimizing the bioefficacy of KRP. 【Method】 L-KRP was prepared by reverse-phase evaporation method, encapsulation efficiency of L-KRP was determined by the fluorescence spectrophotometry, orthogonal experiment was adopted to optimize the prescription using encapsulation efficiency as the evaluation parameter. The appearance, size, Zeta potential, leakage rate and bioactivity of L-KRP were investigated. 【Result】 The dispersed liposome was spherical with the mean diameter of 520.9 nm, the Zeta potential of -83.78 mV and the mean encapsulation efficiency of 77.05%. The toxicity regression equations of L-KRP against the 4th instars of Aedes albopictus were y=3.0499x+0.0368 (r=0.9771), y=2.6003x+1.1592 (r=0.9600) and y=2.7702x+1.2624 (r=0.9765) at different times, with their LC50 were 42.40, 30.00 and 22.35 μg•mL-1, respectively. The permeability of L-KRP was better than KRP through the epidermis of Bombyx mori and Spodoptera litura, the accumulated permeation through B. mori and S. litura were 24.67 and 20.83 μg•mm-2 after 48 hours, with 3.10 and 2.35 times of KRP, respectively.【Conclusion】 All the bioactivity and permeability of L-KRP were higher than KRP.

【Objective】 The impacts of different fertilization regimes on the structure and abundance of soil microbial community were studied.【Method】The influence of fertilization regimes including CK, 1/2MNPK, MNPK, M and NPK on the structure community and abundance of the 16S rRNA gene were analyzed by terminal restriction fragment length polymorphism (T-RFLP) and real-time quantitative PCR.【Result】The sequences from 147 positive clones implied that there were ten phylums bacteria in the soil. They were Firmiutes, Proteobacteria, Chloroflexi, Acidobacteria, Bacteroridetes, Actinobacteria, Verrucomicrobia, Cyanbacteria, Nitrospira, and Planctomycetes. Proteobacteria (relative abundance 26.53%), Bacteroridetes (relative abundance 14.97%) and Actinobacteria (relative abundance 10.88%) were dominated diversities in the soil accounted for relative abundance 52.38%. The highest abundance of Shannon, Simpson’s diversity and Margalef Index were all showed in 0-20 cm soil layer of 1/2MNPK treatment with 3.14, 0.945 and 4.31, respectively. The highest Evenness Index was showed in 0-20 cm layer soil of NPK treatment with 0.941. There were pronounced differences in the primary bacterial community composition and abundance in the different fertilization regimes. RDA results indicated that pH (P=0.002) and soil organic matter (SOM, P=0.006) had the greatest influence on the bacterial community composition. The highest abundance of bacterial both in 0-20 cm and 20-40 cm soil layers were detected in 1/2MNPK treatment. The 16S rRNA gene copy numbers in 1/2MNPK regime with 5.26×109 and 4.96×109 copies/g soil for 0-20 cm and 20-40 cm layers soils respectively, which increased by 90.8% and 197.5% compared to the unfertilized regime.【Conclusion】The dominant populations of bacteria in the soil fertilized with organic manure were significantly different from the chemical fertilizer and unfertilized regime. The rational proportion of chemical fertilizer and organic manure (1/2MNPK) can significantly increase the abundance of 16S rRNA gene in the soil.

【Objective】The difference of nutrient release curves of controlled release urea coated by vegetable oil (PCU) obtained by both in the soil incubation at four growing stages of banana and that in still pure water dissolving incubation was studied in order to prove whether still pure water dissolving incubation method being a simple method to forecast the nutrient supplying rate and amount of the PCU in the soil at different growing stages.【Method】The cumulative nutrients release rate and longevity of the PCU were determined by using soil incubation method taking the still pure water dissolving incubation method as check. The nutrient release characteristics of the PCU at four growing stages during banana growth were measured, respectively.【Result】The nutrient accumulation release curve of the PCU in the soil was different from that obtained by pure water dissolving incubation method. Except for the bud promotion stage the curves at the field seedling stage, the fast growth stage and the young fruit stage changed into “parabolic” pattern from the “sigmoid” one in the water. Within the first 42 days of incubation, the nutrient supply amount of the PCU at all of the three stages was greater while which was smaller at the bud promotion stage and it was in adverse after 42 days. The initial release rate, differential release rate and cumulative release rate of 28 days of the same PCU in the soil was bigger than those in still pure water dissolving incubation, respectively. The initial release rate was in order of the fast growth stage＞the young fruit stage≥the field seedling stage＞the bud promotion stage during banana growth. The differential release rate was in order of the field seedling stage＞the young fruit stage＞the bud promotion stage=the fast growth stage. The volume of cumulative release rate of 28 days was in order of the young fruit stage＞the field seedling stage=the fast growth stage＞the bud promotion stage. Compared with the result measured by still pure water dissolving incubation method, the longevity of the PCU in the soil applied at both of the field seedling stage and the fast growth stage was extended by 18 days and 35 days, respectively, while the longevity at the bud promotion stage and the young fruit stage was shorten by 10 days and 30 days, respectively.【Conclusion】When the same kind of controlled release urea was applied at different growing stages during banana growth, the nutrient supply of the PCU were different at each stage in the same time. The nutrient supply of the PCU was a big amount in prophase when it was applied at the field seedling stage, the fast growth stage and the young fruit stage, respectively, while that was small in later period at those stages. Nevertheless, the nutrient supply of the PCU was a small amount in prophase when it was applied at the bud promotion stage, while that was big in later period at this stage.

【Objective】The objective of the present study is to reveal the genetic diversity, specificity and population differentiation of the North China populations in common apricot (Prunus armeniaca L.). 【Method】 A total of 21 simple sequence repeat (SSR) primers were used to detect the genetic variation, genetic diversity and population structure of 67 common apricots sampled from different regions. 【Result】 All of the 21 SSR loci were polymprphic among the 67 accessions and a total of 301 allelic variations were detected. The average number of allelic variations per locus was 14.33, ranging from 8 to 24. The average Shannon’s information index (I) per locus was 1.934 with a range of 0.65-2.67. By comparison of the diversity among different origins, the genetic diversity of local apricot was rich in the northwest Loess Plateau region. More complementary alleles existed among different geographical origin types and specific alleles existed among different geographical origin types. According to the model-based clustering method, seven groups in the North China populations were detected, and the accessions of different geographical origins were divided into three or more groups. When K=4, all the 63 tested genotypes (expect for kernel-using apricot) were divided into 3 populations: Southwestern group, North China Plain group and Eastern Hills group (Shandong and Southern of Liaoning). This is similar to the traditional classification. 【Conclusion】 A high level of genetic diversity was found in the North China population of common apricot. The most abundant genetic diversity was found in the population from the Northwest. The kernel-using apricot has a narrow genetic base, but has more specific alleles and unique ancestry source. The common apricot of the North China population can be divided into three subgroups, and the accessions from the same origin trends to be divided into the same group, with a few exception.

【Objective】 To provide a theological basis and the basic data on improving saline-alkali soil by natural grass growing techniques in orchard, the effects of this new grass management system on total soluble salt, organic matter, mineral elements concentration, enzyme activity in the soil, and fruit quality in pear orchards in the Yellow River delta were studied. 【Method】 The biochemical attributes of surface (0-20 cm) and sub-surface (20-40 cm) soil in the orchard with golden pear [Pyrus pyrifolia (Burm.f.).cv. Nakai] grown with grass for 2, 4, and 7 years, respectively, were compared with that of the clean cultivated pear orchard and their way of change was analyzed. The items were measured including soil total soluble salt, organic matter, minerals and enzyme activity. The quality of the fruits produced by those orchards was also compared. 【Result】 The results demonstrated that the naturally grown grass significantly increased the organic matter in different layers of the soil from the surface down to 40 cm while effectively decreased the total soluble salt. In pear orchard with 2 years grass growing, the nitrate nitrogen, available phosphate and potassium, exchangeable calcium and magnesium, effective zinc, copper, and ferrous in soil layer of 0-20 cm was significantly lower than that of the clean cultivated orchard, and 7 years grass growing made nitrate nitrogen, ammonium nitrogen, available phosphate and potassium, and effective zinc in the soil layer of 0-20 cm significantly higher than that of the clean cultivated orchard, while nitrogen, available phosphate and potassium, exchangeable calcium and magnesium, effective zinc, copper, and ferrous in the sub-surface soil layer was significantly lower than that of the control. And in the 7 years grass growing orchard, the urease activity in soil layer of 0-20 cm was 0.79 mg•g-1 and alkaline phosphatase was 1.25 mg•g-1, which was 3.8 times and 1.5 times of that of the control, respectively, but the two enzyme activities showed no difference in the layer of 20-40 cm. The fruit fracturability, soluble solids, total aroma, sugars, sugar-acid ratio of pear produced by the orchard with natural grass growing for 4 and 7 years were higher than those from the orchard that had natural grass orchard for 2 years. Correlation analysis indicated that the urease, phosphatase, sucrase, catalase and protease activities mostly showed very significant correlation with N, P, K, Mg and Zn in soil layer of 0-20 cm. The soil enzyme activity mostly showed no significant correlation with all the soil nutrients in layer of 20-40 cm, except the catalase.【Conclusion】 Natural grass could reduce the total soluble salt and increase the organic content obviously in 0-40 cm soil layer of pear orchards in the Yellow River delta. With year increasing, naturally grown grass could improve surface soil enzyme activity, content of mineral nutrition elements and enhance fruit fresh eating quality.

【Objective】The purpose of the paper is to analyze the effect of amylose content in different rice varieties on the qualities of rice vermicelli, and to provide a basis for the selection criteria of suitable rice varieties for vermicelli processing.【Method】Rice vermicelli was processed by one-step molding by using 14 Indica rice varieties whose amylose content ranging from 16.45% to 29.64% as the materials. The processing effect, sensory qualities, cooking qualities and texture properties of rice vermicelli were evaluated, and the correlation between rice amylose content and the qualities of rice vermicelli was established.【Result】With the increase of rice amylose content, the processing effect of rice vermicelli significantly improved. However, when the content exceeds 26.14%, the processing effect declined instead. Chewiness, smoothness, non-stickiness, shininess and overall quality of rice vermicelli showed a significant positive correlations (P＜0.05) with rice amylose contents. A highly positive significant correlation (P＜0.01) was shown between rehydration time and rice amylose contents, and expansion ratio, broken ratio and loss ratio showed highly negative correlations (P＜0.01) with rice amylose content. Texture indexes of rice vermicelli, including hardness, chewiness, shear stress, stretch length, broken force and broken power showed highly significant positive correlations (P＜0.01) with rice amylose content.【Conclusion】The processing effect, sensory quality, cooking quality and texture properties of rice vermicelli are significantly correlated to amylose content of rice varieties. High quality rice vermicelli can be produced by one-step molding processing, using indica rice varieties of amylose content from 22.11% to 25.45%.

【Objective】The objective of this study was to identify the effects of high molecular weight glutenin subunits (HMW-GS) and their combinations on noodle sensory quality, and then select some good HMW-GS and subunit combinations for making high-quality noodles. 【Method】 A total of 80 wheat cultivars were used as test materials to make white dry noodles in lab. The qualities of cooked noodles were evaluated by assessors having been trained and tested with the scaling sensory evaluation analysis method. Analysis of variance (ANOVA) was used to investigate the effect of single subunit at Glu-A1, Glu-B1, Glu-D1 locus and subunit combinations on noodle sensory quality. 【Result】 The results of ANOVA showed that Glu-A1, Glu-B1, Glu-D1 locus had significant influences on the smooth, springiness and total points, hardness and stickiness, respectively (P＜0.1). The influences of single subunit at the locus of Glu-A1, Glu-B1, Glu-D1 on noodle sensory quality were Null＞1, 7+8＞7+9＞14+15, 4+12＞5+10≥2+12. There were significant differences between different subunit combinations for noodle sensory quality (colour, hardness, elasticity, smoothness) and the total score. Wheat cultivars having subunit combination Null/7+8/2+12, 1/7+8/4+12 and 1/7+8/5+10 had higher noodle sensory quality, and had special superiority in some aspects of sensory quality characteristics.【Conclusion】 The HMW-GS of different locus have significant effects on noodle sensory quality. Subunit 7+8 at Glu-B1 locus is the main influence subunit for high-quality noodle. Subunit combinations Null/7+8/2+12, 1/7+8/4+12 and 1/7+8/5+10 are the recommended combinations for noodle used wheat cultivars breeding.

【Objective】 The objective of this experiment is to study the pyridinoline cross-links of collagen and dissect its relevance to the shear force of beef.【Method】 Samples were divided into two groups. The first group consists of the same marbling with different ages. The second consists of the same age with different marblings. The shear force of beef, solubility and hydroxylysylpyridinium, lysylpyridinium of collagen were measured. 【Result】According to the results of the first group, as bull aged, HP cross-links increased, LP cross-links increased firstly, and then decreased, which was followed by the decreasing of collagen solubility and the increasing of shear force. According to the results of the second group, as fattening, HP cross-links decreased, the solubility of collagen increased, and the shear force of beef decreased. 【Conclusion】In the fattening, the synthesization of HP cross-link was restrained. There are shorts of mature cross-links in collagen, resulting in transition of gelatin more easily. So beef turns tenderer.

【Objective】 The objective of the study is to analyze the intrinsic expression bovine fabp3 and fabp4 genes, and the dynamic changes of fabp3 and fabp4 gene expression after transfected the eukaryotic expression vector into mice myoblasts. 【Method】 Eukaryotic expression vector pDsHF3 for fabp3 gene, pDsHF4 for fabp4 gene were constructed and transfected into mice myoblasts by using liposome technique. The transfected mice myoblasts were induced differentiation into myotubes by 2% of pregnant mare serum after 72 h transfection. The changes of gene expression of myoblasts and myotubes were analyzed by real-time PCR. 【Result】 The results showed that, bovine fabp3 or fabp4 genes were expressed in transfected myoblasts, respectively. The intrinsic expression of mouse fabp3 or fabp4 gene was affected in myoblasts and in differentiated myotubes.【Conclusion】 Eukaryotic expression vectors pDsHF3 and pDsHF4 can highly express in proliferative myoblasts, and the expression of exogenous bovine fabp3 or fabp4 genes may affect intrinsic expression of mouse fabp3 and fabp4 genes in myoblasts and myotubes.

【Objective】 Characterization of microRNA expression in hircine ovary tissues was studied, with the purposes of providing basic information for further studies of relationships between specific miRNAs and hircine reproduction such as folliculogenesis and sex hormone secretion. 【Method】 The small RNAs isolated from total RNA of ovary tissues were sequenced by Solexa and then bioinformatics analysis were perfomed. Meanwhile, the expression of selected miRNAs was validated by q-PCR. 【Result】Sequence analysis indicated that 508 ovary miRNAs and 19 corresponding miRNA*s were identified which are conservative in evolution of mammals (sheep, bovine, swine, horse, dog). The expression of 8 selected miRNAs in ovary tissues obtained by q-PCR was in agreement with Solexa sequencing results. 【Conclusion】 The expression profiling of miRNAs, which are abundant and differentially expressed in hircine ovary tissues, was constructed successfully.

【Objective】 The objective of this study is to research the binding function and mode of different odors with general odorant binding protein AcerASP2 of Chinese honeybees (Apis cerana cerana). 【Method】 With the optimization of induction conditions, the purified recombinant AcerASP2 protein was obtained, then the competitive fluorescence assay was used to determine the binding function of AcerASP2 with odors having different structures, finally the homology modeling and molecular docking were applied to elucidate the binding mode and mechanism. 【Result】After the soluble recombinant AcerASP2 protein purified, in the competitive fluorescence assay between AcerASP2 and 1-NPN, the dissociation constants K1-NPN and the number of binding sites n were 7.38 μmol•L-1 and 1.0321, respectively. In selective 7 kinds of odors, the affinity of 4-allylveratrole seemed to be the strongest, and the IC50 and dissociation constant KD were 7.09 and 3.46 μmol•L-1, respectively. The molecular docking results showed that AcerASP2 had one elongated pocket-like hydrophobic cavity, 4-allylveratrole just existed in the cavity, and two hydrogen bonds were found with Lys74 of AcerASP2. 【Conclusion】Because of the particular spatial structure of binding cavity, AcerASP2 could bind diverse odors with different structures, and the binding interaction could be easily to be promoted by the hydrogen bonds between odors with lysines in AcerASP2.

【Objective】 A rapid, sensitive and specific one-step fluorescent quantitative PCR method as a substitute for rabbit fever testing for hog cholera lapinized virus (HCLV) vaccine efficacy was established. 【Method】 A pair of primers and a HCLV specific MGB probe were designed on the 3’UTR region of the HCLV genome for fluorescent quantitative PCR (FQ-PCR). The method was tested for specificity, sensitivity and conformity after optimization. 【Result】 The FQ-PCR sensitivity was 4.35 cDNA copies. The correlation coefficient between CT value and cDNA copies was 0.9998. Amplification efficiency was 101.14%. The FQ-PCR method was specific for HCLV and did not show amplifications for CSFV, BVDV, BDV, PRRSV, FMDV and other pathogens. A total of 34 samples from 17 batches of the four vaccine manufacturers were tested after serial dilutions using the rabbit fever test and FQ-PCR. Eleven samples were disqualified for lack of fever in two rabbits with the Ct values falling between 21.15 and 27.30 and viral content of 8.80×102copy/μL-6.52×104copy/μL. Twelve samples induced fever in one rabbit and no fever in the others with the Ct values between 17.47 and 23.70 and viral content of 1.10×104copy/μL-8.55×105copy/μL. The Ct values of 11 positive samples with both rabbits showing typical fever were from 17.10 to 20.81 with the viral content of 8.27×104copy/μL-1.11×106copy/μL.【Conclusion】The FQ-PCR kits established in this study is specific, sensitive and positively correlated with rabbit fever testing, and thus could be used for quantitative examination of semi-finished HCLV vaccine.

【Objective】To construct postive cell strans inserted the shRNA fragments targeting to porcine J-Domain protein interacting with virus gene, the interference effects were evaluated by comparing the proliferation of classical swine fever virus (CSFV) in constructed cell strains and the cell strain with significant CSFV resistance would be provided as the basic materials for constructing the CSFV-resistant transgenic pig. 【Method】 Four shRNA interfering fragments were designed tagerting to pig Jiv gene and 4 lentivirus inserted the designed shRNA fragment（named P1, P2, P3 and P4）. Four positive PK-15 cell strains transfected with lentivirus were established individually and the interference effects of CSFV were evaluated by determining the RNA of CSFV in positive cell strains after 72 h infected with CSFV by real-time RT-PCR . The porcine fetus fibroblasts were isolated and infected with lentivirus P2 which interfered the proliferation of CSFV remarkerably in cell. A positive porcine fetus fibroblast strain stable expressing the shRNA interfering fragment targeting to Jiv gene was established which also was the donor cells for constructing the transgenic pig. The nuclear of positive pircine fetus fibroblast was transferred into matured enucleated porcine oocytes, and somatic embryos were obtained and transplanted into the receptor sows for constructing transgenic pig. The foreign gene was identified in obtained transgenic pig.【Result】 Four PK-15 cell strains inserted into the shRNA interfering fragments targeting to the Jiv gene of porcine were established, the P2 cell strain (inserted into P2 interfering fragment) was significantly CSFV-resistant. The nuclear of porcine fetus fibrlblast inserted P2 interfering fragment was donor cells for constructing transgenic pig by somatic cell cloning and embryo transplantation, and a transgenic piglet was obtained.【Conclusion】 The expression of Jiv gene can influence the proliferation of CSFV in cells. A PK-15 cell strain with stable expressing shRNA P2 was established which could resist CSFV proliferation remarkablly. A trnasgenic piglet was constructed by somatic cell cloning and embryo transplantation which carries the P2 interfering fragment targeting to porcine Jiv gene.

【Objective】 The classical swine fever (CSF) C-strain vaccine has proven to be safe and highly efficacious. However, classical swine fever virus (CSFV) genotype Ⅱisolates became dominant in recent years, making it more challenging to bring the disease into total control. This study was attempted to engineer, by reverse genetic approach, a novel C-strain-based vaccine candidate that matches molecular variations of field isolates, and to characterize its biological features. Those results could provide foundations for further development of a marker vaccine for differential diagnostic purpose.【Method】An infectious recombinant CSF virus was generated by exchanging the 5’-end 870-bp of E2 gene in C-strain with the same fragment from group 2 strain HZ1-08. The resulting recombinant virus RecCHZE2 was rescued and its pathogenicity and immunogenicity were characterized in rabbits and pigs. 【Result】 Rabbits inoculated with RecCHZE2 exhibited typical temperature profiles similar to the C-strain with spleen enlargement. Pigs inoculated with the recombinant chimeric virus did not show fever and significant decline of the number of white blood cells. Mild transient viremia was detected at day 12 post-inoculation. The antibody responses specific to the field strain HZ1-08 reached the peak 20 days post-inoculation. The neutralizing antibody titers from pigs immunized with RecCHZE2 were similar to both HZ1-08 and C strains while the titer from pigs immunized with RecC were lower for group 2 strain HZ1-08.【Conclusion】The recombinant virus RecCHZE2 remains avirulent but differs in immunogenic properties from the C-strain.

【Objective】Vaccination is still the main strategy for CSFV control in China. CSFV HCLV(Hog Cholera Lapinized Virus, HCLV) strain could protect effectively, but it couldn’t be serologically differentiated between infected pigs and vaccinated pigs (DIVA principle), which could been effectively solved by marker vaccine and marker diagnostics.【Method】Based on infectious cDNA clone of CSFV T strain, which is a temperature-sensitive mutation strain, the complete structural protein-coding region of T strain had been replaced by that of ShiMen strain , and the recombinant plasmid SMT was constructed. Additionally, the Flag gene was inserted into C end of E1 protein of the recombinant virus as a positive marker, and the pSMT-Flag was constructed successfully. After electroporation, the recombinant viruses vSMT and vSMT-Flag were passaged by SK6 cell. 【Result】Two recombinant plasmids were identified by specific restriction enzymes，the rescued viruses could be passaged successfully and confirmed by RT-PCR, sequencing and immunoperoxidase monolayer assay(IPMA). 【Conclusion】 This marker virus could react with the Flag specific monoclonal antibody while could not influence the combination between E2 protein and its specific monoclonal antibody, which could be a potential CSF novel marker vaccine.

【Objective】The objective of this study is to clone a novel gene of MYB family (GaMYB2) from Gossypium arboreum L., to analyze its expression patterns, and to characterize the deduced protein of this gene. 【Method】The full length of GaMYB2 was cloned by RACE and RT-PCR techniques, the gene sequence obtained and the putative amino acid sequence was analyzed by bioinformatics software. Its expression patterns were analyzed by real-time PCR under normal and PEG6000 stress respectively，which simulated drought tolerance.【Result】The GaMYB2 full length cDNA sequence is 1 117 bp, and the ORF is 840 bp, which encode a deduced protein including 279 amino acid residues. Blastp reseach revealed 40.50% to 68.20% similarities between GaMYB2 and other reported species, such as Oryza sativa (BAA23338.1) and Triticum aestivum (AAT37168.1). Subcellular localization showed that it expressed possibly in cell nucleus. The experiment results of real time PCR exhibited that GaMYB2 could be induced by 17% PEG6000, and had a higher expression level in root and flower than in other tissues. 【Conclusion】The GaMYB2 , cloned for the first time from G. arboreum L., is a new transcription factor of MBY family, and may play an important role in plant tolerance to drought stress.

【Objective】 To study the function of BnICE1 in response to low temperature, transcription factor BnICE1 which is involved in cold stress responsiveness in Brassica napus was cloned and its sequence was analyzed. 【Method】 The full-length cDNA of BnICE1 was obtained by the technology of rapid amplification of cDNA ends (RACE). The cDNA sequence and the deduced amino acid sequence were analyzed by bioinformatics method. The spatial-specific expression of BnICE1 in different tissues of the control plants and the expression patterns of BnICE1 in leaves under cold stress condition were determined by real-time RT-PCR. 【Result】In this study, a novel gene, BnICE1, from B. napus was isolated and characterized. The full-length cDNA was 1 737 bp, containing a 1 500 bp opening reading frame(ORF). The deduced protein was 499 amino acids with molecular weight 53.2 kD and isoelectric point 5.0. The sequence aligment demonstrated that the C-termina1 of deduced BnICEI protein exhibited a typical bHLH domain and was high identity with other ICE1 proteins from other species. So it was named the BnICE1 (GenBank accession No. JF268687). The result of phylogenetic tree showed that BnICE1 was closer to BcICE1 and BchICE1. Analysis by real-time RT-PCR indicated that BnICE1 was expressed in different tissues (stem, leaf, hypocotyls) and that relatively higher abundance was observed in hypocotyls. 【Conclusion】 The transcription factor BnICE1 was cloned from B. napus and the analysis by real-time RT-PCR suggested that BnICE1 played an important role in defending cold stress.

【Objective】 The dynamic changes in leaf curve of rice under different nitrogen levels were analyzed using Gaussian function. 【Method】Based on rice pot experiments involving different years and nitrogen levels, a 3-D laser scanner was used to measure the spatial coordinate data of leaf curve at different leaf positions on main culm of rice under different nitrogen levels, then dynamic modeling technology was used to build the model of leaf curve changes at different leaf positions on main culm in rice. 【Result】 The results indicated that the angle between the end of leaf curve and positive direction of y-axis (AH) increased with growth degree days on the trend of slow-fast-slow, which could be described with the Logistic function. From the first leaf to the seventh leaf, the maximum value of AH increased with increasing leaf position, and then decreased with increasing leaf position. The significant linear relationship could be observed between sheath-leaf angle∠BFC and AH. The logistic equations were used to describe the changes of AH and the model parameter Sm in leaf curve model with growth degree day (GDD), respectively. The piecewise function was applied to describe the change of maximum AH with leaf position. A variety parameter (AH of the seventh leaf position on main culm in rice under optimal nitrogen conditions) and nitrogen factor were introduced to quantify the effects of variety and nitrogen rate on leaf drooping degree, respectively. Model validation with independent field experiment data showed that the average value of LTS-HD (least trimmed squares Hausdorff distances) between observed and predicted curve are less than 0.88 cm and 1.18 cm at tilling and jointing stages, respectively. 【Conclusion】 The model showed a good predictability for spatial dynamic change of leaf curve on main culm under different nitrogen rates during main growth stages in rice. These results would provide a technical support for visualization of leaf and plant in rice.

【Objective】 This study was conducted to determine the allelic constitution and subspecies attribution of the super hybrid rice Yiyou 673, so as to facilitate the utilization of beneficial alleles carried by Yiyou 673, and to provide information for establishing an appropriate indica-japonica proportion in rice breeding. 【Method】 Three major restorer lines of the three-line hybrid rice, including the founder parent Minghui 63, its derivative Minghui 86, and Fuhui 673 which is derived from Minghui 86, as well as cytoplasmic male sterile line Yixiang A, and Yiyou 673 which is developed by crossing Fuhui 673 to Yixiang A, were assayed with 17 markers for 13 genes of agronomically importance and 15 SSR markers for indica-japonica differentiation. 【Result】 Polymorphism was detected in eight of the gene marker regions, including two genes for heading date and yield traits, two key genes for grain quality, and four genes for blast resistance. At Wx locus which is responsible for amylose content and gel consistency of the milled rice, a high-quality allele was transmitted from Minghui 63 to Fuhui 673 through Minghui 86. At ALK locus controlling gelatinization temperature and two loci for blast resistance, Pib and Pita, the male and female parents of the F1 hybrid rice Yiyou 673 showed allelic complementation for high grain quality and resistance to blast. In addition, it was shown that Minghui 63, Minghui 86 and Fuhui 673 are all indica-inclined, but the japonica component of 40.0% in Minghui 86 and Fuhui 673 is higher than the value of 33.3% in Minghui 63.【Conclusion】 Beneficial alleles derived from Minghui 63 and other parental lines were pyramided in Fuhui 673, which were combined with those carried by Yixiang A to establish a favorable allelic complementation in Yiyou 673.

【Objective】One hundred and twenty-seven accessions of kenaf gemplasm collected from different countries and areas, which contain cultivated, wild and related species, were selected and SRAP markers were used to establish molecular identity of kenaf germplasm.【Method】 Sequence- related amplified polymorphism(SRAP) marker was used for analyzing genetic relationships of the 127 different type accessions. Nei's similarity coefficients were generated and dendrogram was constructed using UMPGA by NTSYS-PC program.【Result】Three hundred and eighty-three bands were amplified by 40 SRAP primers, of which 375 were polymorphic and PPB was 97.9%. The cluster analysis based on UPGMA showed that the 127 accessions of kenaf germplasm were divided into four major groups at similarity coefficient of 0.70. All of the accessions could be effectively distinguished by specific bands and different primer combinations. The specific molecular identity for 127 accessions of kenaf germplasm were established with 99.99% probability of confidence.【Conclusion】A set of unique molecular identity was established for 127 kenaf germplasm based on 36 bands which were amplified by 15 core primer combinations.

【Objective】 The physiological indices of substitution lines in different phosphorus treatments were studied to select species of tolerance to low phosphorus stress. 【Method】 The comprehensive tolerance to low phosphorus stress of low phosphorus tolerance coefficient (LPTC) of soluble sugar, soluble protein content, SOD activity, POD activity, MDA content, leaf chlorophyll content, corticoid content, Pn, Tr, Gs, Ci, Fv/Fm of Chinese Spring (CS) - Synthetic 6x substitution lines as well as their parents were evaluated by principal components analysis and cluster analysis. 【Result】 The results showed that the 14 single indexes could be classified into 7 independent comprehensive components. The cluster analysis was used to divide the 21 substitution lines and their parents into three types of tolerance to low phosphorus. 2A substitution line and Synthetic 6x belonged to high tolerance types to low phosphorus stress; Chinese Spring and 4A, 7A, 2B, 1D and 3D substitution lines belonged to poor tolerance types to low- phosphorus stress. The others including 1A, 3A, 5A, 6A, 1B, 3B, 4B, 5B, 6B, 7B, 2D, 4D, 5D, 6D, and 7D substitution lines belonged to middle tolerance types to low- phosphorus stress.【Conclusion】 The comprehensive evaluation and principal component analysis provide a useful way for selecting species of tolerance to low phosphorus stress in wheat.

【Objective】The effects of soil properties and vegetable yield under different intercropping pattern treatments of waxy maize and maize straw returning were studied. Continuous cropping was used as a contrast. The aim was to supply academic base for solving continuous cropping obstacle and summer fallow problems in vegetable field. 【Method】 Broccoli, green sword bean and waxy maize were used as materials, and three treatments were set up. There were relay-intercropping mode of green sword bean and waxy maize, relay-intercropping mode of green sword bean and waxy maize with waxy maize straw returning, broccoli, green sword bean continuous cropping. The soil samples were used for chemical analysis after natural drying. Soil organic matter content, soil NH4+-N content, soil NO3--N content, the activity of urease, the activity of soil polyphenol oxidase and the activity of soil catalase were analyzed. At harvest period, the yields of broccoli, green sword bean and waxy maize were determined. 【Result】In the year 2010, the autumn vegetable yield of the relay-intercropping treatment was the highest, the autumn vegetable yield of the continuous cropping was the lowest. The principle was in agreement with the autumn vegetable yield of 2011. The results showed that soil organic matter content and soil urease activity were significantly increased under relay intercropping treatments and waxy maize straw returning treatment by using nutrients reasonably, compared with the vegetable continuous cropping. Intercropping or relay cropping decreased the inorganic nitrogen concentration, and increased the inorganic N utilizing rate in vegetable fields compared with sole cropping. Relay intercropping and waxy maize straw returning mode promoted microorganism propagation and made the soil urease activity at the high level. Vegetable continuous cropping caused soil nutrient over consumption and decreased the activity of urease. As more toxic substances from the root was released to the soil, this phenomenon made that the activity of soil polyphenol oxidase and catalase in vegetable continuous cropping fields were higher than that in other treatments, the increasing of the CAT activity will reduce the toxicity of the soil.【Conclusion】The results of the experiment indicated that crop yield increased effectively under relay-intercropping and straw returning. At the same time, relay-intercropping improved soil environment, organic matter content and urease activity increased, and the activity of soil polyphenol oxidase and catalase remained at an appropriate level.

【Objective】 This experiment was carried out to establish an efficient anther culture system of mutlifoliate alfalfa, and get regenerate plantlet. It will also provide basic materials for the research of ploidy, genomics and molecular biology of alfalfa.【Method】 Firstly, Huaiyin alfalfa and a mutlifoliate alfalfa varieties named Grandeur were used as the parents to create a hybrid, and the anther of hybrid F1 was used as the explants. The ability of induction of callus after bud was studied by using the culture medium of NB added with 2,4-D (0.5 mg•L-1), NAA (0.3 mg•L-1) and KT (3.0 mg•L-1). The appropriate medium was screened for the differentiation of anther callus of alfalfa by orthogonal design L16 (45). In addition, the rooting ability from buds was studied by adding different concentrations of NAA into 1/2 B5 and 1/2 MS.【Result】The highest induction rate of alfalfa callus was in 15 to 28 days after budding, and the maximum value was 82.79%. The culture medium that was good for the differentiation of the callus of alfalfa anther was made up of BⅠwith CH (200 mg•L-1), 2-IP (0.5 mg•L-1), NAA (0.2 mg•L-1) and KT (1.0 mg•L-1), and the maximum regeneration rate was 54.05%. The best culture medium for adventitious bud was 1/2 B5 with NAA (0.3 mg•L-1), and the maximum rooting rate was 88.46%.【Conclusion】An efficient anther culture system of mutlifoliate alfalfa was established successfully, and the regenerated plants were obtained.

【Objective】The objective of this study is to isolate and identify the lipopeptides produced by Bacillus amyloliquefaciens Lx-11 and their biocontrol efficacy against bacterial leaf streak (BLS). 【Method】 Three primers designed based on the known lipopeptide genes were used to amplify the corresponding genes from the genome of Lx-11 strain. Lipopeptides were identified by MALDI-TOF-MS techniques.【Result】The PCR products with three primer pairs were cloned and sequenced. The BLAST analysis showed that the sfp, fenB, ituA or bamA genes exist in the genome of B. amyloliquefaciens Lx-11. Three kinds of lipopeptides surfactin, bacillomycin D and fengycin were identified in the antimicrobial extract by the mass spectrum analysis. The antibacterial activity could be associated with surfactin-lipopeptides, and practically abolished in surfactin-deficient mutants. Lx-11 strain significantly reduced disease incidence of BLS.【Conclusion】The antibacterial activity of the surfactin- deficient mutant strain was practically abolished, which suggested that surfactin might play an important role in control of BLS.

【Objective】The objective of this study is to isolate NBS-LRR class resistance gene homology fragments from wheat using homology-based method, and to validate the relationships with wheat stripe rust resistance gene Yr26.【Method】Primers were designed based on the conserved domains of the cloned plant disease resistance genes, and the nested PCR was used. The fragments were localized by Chinese Spring (CS) deletion lines of chromosome 1B, F2:3 populations derived from susceptible cultivar Avocet S and the lines 92R137 carrying Yr26 were used to evaluate the relationship between the isolated RGAs and Yr26. 【Result】Four open-reading resistance gene analogues (RGAs) were obtained, i.e., R105, R181, R326 and R405. The nucleotide sequences of the four RGAs were 369, 589, 528 and 618 bp. Homology research showed that the four fragments had typical conserved regions NBS-LRR, and the nucleotide identity of four fragments was from 24.35% to 38.36%. Only R405 was mapped in the deletion bin C-1BL-0.6-0.32 with Chinese Spring deletion lines and was co-segregated with a F2 population of 196 plants. It was initially hypothesized that the R405 might be the candidate sequence of Yr26.【Conclusion】In this study, four resistance homology fragments were obtained in the near isogenic lines (NILs) Nan137. R405 was mapped in the Yr26 region and co-segregated with Yr26, which provides a good prerequisite for further study.

【Objective】In the genus of Verticillium, there are five ‘species’ as plant pathogens which could lead to plant wilt symptom. The aim of this study was to identify the specific ‘species’ caused Verticillium wilt of cotton.【Method】A total of 310 Verticillium isolates were obtained from 84 counties, 12 provinces of China. To clarify the ‘species’ of them, both observation on biological characteristics and molecular identification were used.【Result】A total of 298 isolates were typical V. dahliae producing black microsclerotia and verticillate conidiophores identified by specific PCR amplifications. However, 12 special isolates producing non-microsclerotia, black mycelia or chlamydospore were tested by temperature sensitivity, specific PCR amplification and ITS sequencing. Except for 2 V. nigrescens isolates, the rest 10 isolates were identified as V. dahliae, no V. albo-atrum was found. 【Conclusion】 In 3 main cotton growing regions of China, the causal agent of cotton Verticillium wilt is V. dahliae. And V. nigrescens also presented in Yangzi River and Yellow River basins. In cotton Verticillium wilt happening process, the interaction between V. dahliae and V. nigrescens needs to be further studied.

【Objective】 The effects of nicotine on the feeding behaviors of the Bemisia tabaci B and Q biotypes were investigated, with the aim of establishing a corresponding evaluating system based on the electrical penetration graph (EPG) and the liquid diet sac (LDS) technique. 【Method】 EPG method combined with a LDS was used to quantify the effect of the concentrations of nicotine on the feeding behaviors of the B. tabaci. 【Result】 Relative amplitudes and frequencies in EPG waveforms of the B. tabaci in 10% sucrose solution, compared to those in 5% and 15% sucrose solutions, were analogous to the waveforms of B. tabaci on plants. The durations of ingestion waveforms decreased with the increase of nicotine levels in 10% sucrose solution. Salivation (E1) and passive ingestion (E2) durations were reduced and the penetration duration was prolonged even at the low concentration (0.001%) of nicotine. The passive ingestion from LDS was completely inhibited at a higher concentration of nicotine (0.01%). Although the major EPG pathway variables between B. tabaci B and Q biotypes were similar, the duration of passive ingestion of Q biotype was significantly longer than that of the B biotype on the diet without nicotine or with lower nicotine concentration. 【Conclusion】 The valuable EPG variables for EPG+LDS system include the durations of C, E1 and E2 patterns. Ten percent sucrose solution in LDS was more suitable for recording the feeding behaviors of B. tabaci using EPG technique. The feeding behaviors of B. tabaci B and Q biotypes were signi?cantly inhibited by nicotine, but Q biotype had higher nicotine adaptability than B biotype.

【Objective】 The effects of carbonized maize stalks on the brown soil phosphorus fractions and availability were studied to ascertain whether or not the carbonized maize stalks is good for soil phosphorus fertility development.【Method】 Laboratory culture technique was used to compare the difference among the carbonized, non-carbonized and decomposed maize stalks treatments (same quantity carbon application in different maize stalks treatments), and the chemical fertilizers treatment alone with same amount application of chemical fertilizer. The fractions of soil phosphorus were determined by the modified Hedley’s sequential extraction procedure. The adsorption and desorption isotherms for soil phosphorus were determined by the method of Yan Chang-Sheng with Constant temperature shaking. 【Result】 The total phosphorus content in the soil could be significantly improved by all kinds of maize stalks treatments compared with the no maize stalks (single chemical fertilizer) treatment, and the phosphorus contents increased were significantly correlated with the quantities of maize stalks added into the soil. The soil phosphorus fractions could be affected significantly by the added different maize stalks. Although the available phosphorus content was not high in the carbonized maize stalks, but the sodium bicarbonate-extractable inorganic and organic phosphorus contents could be significantly improved by application of the carbonized maize stalks. The adsorption amount of phosphate ions could be reduced significantly by the maize stalks treatments, and the impact of carbonized maize stalks treatment was the most obvious. Whether the maize stalks was decomposed or not, the change of the phosphorus adsorption quantities was not obvious. 【Conclusion】 The application of carbonized maize stalks can promote the soil phosphorus availability, reduce soil phosphorus fixations, and improve the utilization ratio of the phosphorus fertilizer, but the increasing activity of soil phosphorus may also increase the phosphorus environmental risks.

【Objective】To provide a theoretical basis for balanced fertilization of rapeseed production in the middle of Sichuan hilly regions, abundance and deficiency indexes of soil nutrient and fertilization recommendation indexes were established. 【Method】Data were analyzed based on the “3414” experiments of rapeseed in the field from 2005 to 2009. Making use of plentiful-lack of soil nutrient as an index, soil available phosphorus (AP) and available potassium (AK) were divided into five levels on the basis of different relative yields. Nitrogen fertilizer application rate (N) on average was estimated based on the method of total quantity control, while phosphorus fertilizer (P2O5) and potassium fertilizer application rates (K2O) were calculated by the method of fertilizer response function. 【Result】 Regional optimal application rate of chemical fertilizer N on average is (170±50) kg•hm-2. Phosphorus fertilizer application rates (P2O5) are 100-120,70-100, 50-70, 30-50, ＜30 kg•hm-2 (AP＜5, 5-15, 15-25, 25-45 and ＞45 mg•kg-1), while potassium fertilizer application rates (K2O) are 90-110, 70-90, 50-70, 30-50, ＜30 kg•hm-2 (AK＜30, 30-60, 60-120, 120-240 and ＞240 mg•kg-1). 【Conclusion】Compared with the fertilization recommendation rate calculated by the original system, the fertilization recommendation rate above suggests that nitrogen and phosphorus fertilizer application rates should be decreased, while potassium fertilizer application rates ought to be increased.

【Objective】 Soil nutrient fertility status was investigated to formulate a suitable fertilization strategy so as to help to raise its economic and environmental benefits. 【Method】 Soil samples from different layers (0-20 cm, 20-40 cm and 40−60 cm) were collected from 99 vineyard producing areas located in Changli, Huailai, Xuanhua and Zhuozhou counties in Hebei Province, respectively. 【Result】The content of soil organic matter was low at large, the deficiency rate of 0−20 cm, 20−40 cm and 40−60 cm was 52%, 76% and 90%, respectively. Moreover, the CV of vineyards soil N, P and K was high and the CV of surface soil was 113.1%, 143.5% and 68%, respectively. Similarly, the concentrations of Fe and Cu in vineyards soil were too high, 94% and 99% in surface soil above suitable content, respectively. However, the contents of Zn and Mn were low and the deficiency rate of Zn and Mn was 56% and 100% in surface soil, respectively. 【Conclusion】It was suggested that more organic matter, Zn, Mn, rational application of nitrogen and less Cu fungicide should be used in the tested vineyards.

【Objective】 The aim of the experiment was to explore the effects of exogenous NO donor (SNP) and abscisic acid on pepper resistance ability of autotoxicity. 【Method】 Pepper cultivar ‘Longjiao 2’ was used as materials in this experiment to investigate the effects of SNP (150 μmol•L-1) and ABA(100 μmol•L-1) on gas exchange parameters, cholorophyll fluorescence parameters and chlorophyll content in pepper seedlings under autotoxicity stress. The seedlings of pepper were treated with soil and substrates extracts of 3 years continuous cropping pepper. 【Result】 It was found that all of the soil and substrates extracts reduced Pn, Gs, Tr, Fv/Fm, ΦPSⅡ, qP, chlorophyll content and ratio of chlorophyll a/b obviously, but increased Ci and NPQ significantly. Under autotoxicity stress, exogenous NO treatments significantly increased Pn, Gs, Fv/Fm, ΦPSⅡ, qP and chlorophyll content, but reduced Ci and NPQ significantly, and had no obvious influence on Tr. Exogenous ABA treatments significantly increased Pn, Fv/Fm, ΦPSⅡ, qP and chlorophyll content, but reduced Ci and NPQ significantly, and had no obvious influence on Gs and Tr under autotoxicity stress.【Conclusion】The reduction in net photosynthesis rate caused by autotoxicity stress was considered to be a result of non-stomatal restriction mainly, and autotoxicity on pepper treated with soil extracts was serious greatly than substrates extracts. Under autotoxicity stress, exogenous NO and ABA application increased chlorophyll content, maintained PSII center activities and photosynthetic capacity, and enhanced autoxicity tolerance in pepper.

【Objective】 In order to provide technical support for molecular marker-assisted breeding for Fusarium wilt resistance in watermelon, molecular markers tightly linked to Fon-1 for Fusarium oxysporum f. sp. niveum race 1 resistance were developed.【Method】An F2 segregating population derived from a cross between the cultivated resistant male parent ‘Calhoun Gray’ and the susceptible female parent ‘Black Diamond’. The bulked segregant analysis (BSA) was employed to map the Fon-1 gene interval preliminaryly. A series of candidate single nucleotide polymorphism (SNP) sites that are linked to the target traits were obtained, and resequencing analysis within the target gene interval among 11 cultivated watermelon accessions was conducted. And then, the cleaved amplified polymorphic sequence (CAPS) or derived cleaved amplified polymorphic sequence (dCAPS) markers were designed to verify the linkage relationship between these CAPS/dCAPS marker and Fon-1 gene using F2 segregating population and germplasm resources. 【Result】 The preliminary results from screening of the resistant and susceptible DNA pools with 950 pairs of SSR/Indel/dCAPS primers mapped the Fon-1 gene to a 15 cM interval of watermelon chromosome 1. Three markers, 7716_fon, 7419_fon and 4451_fon, which were demonstrated to be tightly linked to Fon-1 gene, were developed using the SNP sites confirmed in the F2 population and 164 cultivars and the genetic distance between the markers and the Fon-1 gene was estimated to be 0.8 cM, 1.0 cM and 2.8 cM, respectively.【Conclusion】Three CAPS/dCAPS markers that can effectively distinguish the resistance/susceptibility to Fusarium oxysporum f. sp. niveum race 1 in cultivated watermelon were developed in this study and these markers will provide useful technical support for molecular breeding for resistance against F. oxysporum f. sp. niveum race 1 in cultivated watermelon.

【Objective】Duck RIG-I (duRIG-I) gene was cloned and the functions of its different domains were analyzed preliminarily. 【Method】 The CDS of duRIG-I gene was cloned on the basis of the sequence submitted to GenBank with RT-PCR and was analyzed by bioinformatics. The eukaryotic expression vectors of N-terminal, C-terminal and whole-length of duRIG-I gene with 6*his tags were constructed to transfect DF1, and then the transcription and expression of the three recombinant plasmids in cells were detected via RT-PCR and indirect immunofluorescent assay, respectively. Meanwhile, the expressions of chicken IFN-β, Mx1 and PKR mRNA were detected by real-time PCR.【Result】The whole-length of duRIG-I CDS was 2 802 bp encoding 933 amino acids. All the recombinant protein of duRIG-I could express normally in DF1. The results of RT-qPCR indicated that CARDs significantly up-regulated the mRNA level of IFN-β, Mx1 and PKR genes.【Conclusion】The various domain fragments of duRIG-I express normally in DF1. The N-terminal of duRIG-I plays a vital role in regulating the expression of downstream genes of RLR antiviral signal pathway.