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China is the largest rice producer worldwide and is one of the origins of Asian cultivated rice as well. Along with the rapid development of transgenic rice in China, the potential impact of rice transgene flow on the environment and food safety has become one of the major concerns. Gene flow is an important parameter in the risk assessment and regulation of transgenic rice on the scientific basis. In accordance with this situation, we have formed a team and systematically studied the rice transgene flow since 2002. The results obtained in recent ten years are as following: (1) the patterns of transgene flow and the major biological and meteorological factors controlling rice gene flow have been elucidated. Following the prevailing wind direction in rice flowering period, a rectangular design of field experiments were conducted at 3 locations（Sanya, Hainan Island; Guangzhou, Guangdong; and Hangzhou, Zhejiang）in 2-3 years by using a homozygous transgenic line L201 or B2 (sister lines) with bar gene inserted, resistant to herbicide Basta, as a pollen donor, and totally 19 non-transgenic rice as recipients, including male sterile (ms) lines, common rice cultivars (CRC), F1 hybrid rice, and common wild rice (Oryza rufipogon). Results indicated that the frequency of transgene flow to ms lines was the highest, while gene flow to CRC and F1 hybrids was the lowest (less than 1% or 0.1% at parallel plantation). The frequency of transgene flow to O. rufipogon was in between. By comparison, the maximum frequency of gene flow to ms lines is one to three orders of magnitude higher than that to O. rufipogon and CRC. Gene flow frequency decreased exponentially as the distance increase, with a sharp cut-off point at about 1-2 m in Guangzhou and Hangzhou, while it was approximately 5m in Sanya. It indicates that the sharp cut-off point is closely related to the wind speed during rice flowering period at a given location. By using a concentric circle design of field experiment and an ms line BoA with higher outcrossing rate as a recipient, we have been able to clearly quantify the relationship between the gene flow frequency and the wind direction. On the basis of the cumulative data in eight compass sectors, 90%–96% of the cumulative gene flow events occurred in the four downstream prevailing wind sectors, while it was only 4%–10% in the four lateral and upstream prevailing wind sectors. In short, a general conclusion is that the order of magnitude of transgene flow frequency is basically the same as the outcrossing rate of the CRC (generally less than 1%), which means the gene transfer has not added a new additional risk. (2) By using historical meteorological data as an input, a regional applicable model of rice pollen dispersal and gene flow has been established, which is successively used to predict the maximum threshold distances (MTDs) of gene flow in 17 provinces of southern rice growing area in China. The feature of spatial distribution of MTDs shows: from east to west, MTDs gradually decrease; from north to south, MTDs first decrease in the hilly region and then increase again along the southeast costal region. Reason for it is that the spatial distribution is dramatically influenced by the southeast monsoon (seasonal wind direction from the southeast) and the landform structure. (3) We have artificially constructed two mixed populations of O. rufipogon with F1 hybrids of CRC/O. rufipogon derived from transgene (either Bt or bar) flow to investigate the long-term fate of the transgene integrated into common wild rice. It was found that the F1 hybrids of CRC/O. rufipogon totally disappeared within 3-5 years and the Bt or bar gene was not detectable in the mixed population. It is reasonable to speculate that the common wild rice possesses a mechanism of self-protection. (4) The effectiveness in reducing transgene flow by using flowering isolation or a protective cloth-screen in small-scale field trials of transgenic rice was studied and the results discussed. To investigate the degree of flowering synchronization of CRC and O. rufipogon populations in adjacent plantation, a survey has been conducted in Hainan, Guangdong and Guangxi provinces which allow us to establish a corresponding database. In order to further eliminate the rice transgene flow, a biological containment measure - gene split approach has been established. Based on the data obtained in this study and a survey on the internationally published data of gene flow from major crops, we have proposed to use principles of classification management and threshold-value management in the risk assessment and regulation of transgenic rice. Meanwhile, the progress and prospective of rice gene flow are also discussed in this paper.

【Objective】 The objective of this study is to analyze the genetic diversity of the three-line hybrid rice parents based on functional genetic markers. 【Method】 Genetic diversity of 76 three-line hybrid rice parents was analyzed by 44 functional gene markers involved in QTL loci, or fine mapping, or been cloned which linked closely to rice yield trait, and these genes have been considerably reported by literature. At the same time, the genetic diversity of the above materials mentioned was also studied by 29 SSR markers with higher polymorphism, clear band pattern, reproducible, and covered in 12 chromosomes of three-line hybrid rice parents. According to claim of POPGEN32 analysis software, the PCR gel electrophoresis product data matrix was transformed into the genotype data, and the alleles (Na), effective number of alleles (Ne), percentage of polymorphic loci (P), and Nei’s genetic diversity index (He) were calculated. This study further evaluated the genetic diversity of all parents according to the deduced information of POPGEN32 analysis software. Besides, this study also calculated the genetic differentiation coefficient (Fst), Nei’s genetic distance (D), and further checked out the genetic structure and genetic relationship. NTSYS-pc2.10e software was used to calculate the genetic similarity coefficient (GS), and cluster analysis was made according to GS group and using the non-weighted average method (UPGMA), and the genetic dendrogram was mapped. 【Result】Of which 37 functional gene markers showed polymorphism and 86 total alleles loci were detected; the percentage of polymorphic loci (p) was 84.09%. While the number of effective alleles(ne) was 62.95, which accounted for 73.2%, Nei’s genetic diversity index(he) ranged from 0.049 to 0.831, and 0.650 in avarage. The genetic similarity (GS) of 76 varieties ranged from 0.323 to 0.973, and 0.650 in average. UPGMA cluster analysis showed that 76 accessions could be classified into two distinct classes of maintainer lines and restoring lines, at similarity coefficient of 0.618. The coefficient of genetic differentiation(gst) was 0.151, belonging to high variation level, and the Nei’s genetic distance (GD) was 0.185. The genetic distance within the groups was relatively small, relatively large among the taxa. A total of 72 alleles loci were detected by 29 SSR markers. UPGMA cluster analysis showed that the 76 accessions cannot be classified into two distinct classes of maintainer lines and restoring lines. Part maintainer lines clustered in a group of restoring lines, some restoring lines clustered in the maintainer line group. 【Conclusion】 The studies suggested that the functional gene markers had a high DNA polymorphisms detection efficiency, and can be used as a useful tool for their accuracy and reliability of measuring genetic diversity. The backbone parents in research showed nearer genetic relationship, higher homology sort of genetic basis. However, there still showed higher genetic differentiation between maintainer line and restoring line, suggesting that there was a higher space of using heterosis breeding in the yield of rice parents.

【Objective】Rapeseed oil contains many fatty acid components and the major goal in breeding and genetic research is to increase of the oleic acid content and decreases of the linolenic acid and erucic acid content. QTL mapping of oil quality related oleic acid, linolenic acid and erucic acid content in Brassica napus was made by using the high density SNP genetic map which was constructed from the high generation RIL population with the newly developed rapeseed 60K chip array. 【Method】 The high generation RIL population was derived from the cross of high erucic acid female parent GH06 and low erucic acid male parent P174. Seeds harvested from Giessen and Hohenlieth in Germany in 2008 and again in Giessen in 2009 were measured by NIRS with three technical repeats. The RIL population genotype was analyzed with the newly developed rapeseed 60K chip array. The DNA preparation and the chip preparation were processed strictly according to Infinium HD Assay Ultra manual of Illumina Inc. The SNP linkage map was constructed by using MSTmap program with minimum LOD 5.0. The reference SNP genetic map contains 2756 SNP markers, covering 1 832.4 cM of B. napus genome. QTL mapping of oleic acid, linolenic acid and erucic acid content was conducted by composite interval mapping using software Windows QTL Cartographer. 【Result】 Trait analysis indicated that the oleic acid and erucic acid contents were controlled by major genes, and their correlation coefficient was higher than -0.95 in the three different environments at significant level. Linolenic content showed a negative and positive correlation with erucic acid and oleic acid content, respectively. Two major QTL located on A08 and C03 were detected both for oleic acid and erucic acid content at the same position. In the three different environments, the major oleic acid QTL accounted for the phenotype variation from 19% to 31%, and the major erucic acid QTL effects varied from 19% to 34%, and the two major QTL for oleic acid and erucic acid content gave opposite direction of additive effect. The additive effect of the major erucic acid QTL on A08 and C03 come from the low oleic acid and high erucic acid parent GH06 with value varied from 7.6 to 9.6 in three different environments. Different QTL for linolenic acid was found across three different environments with QTL effects varied from 3% to 12%, which was coincided with that the linolenic acid was a typical quantitative trait, and was easily affected by the environment factors. The genetic map and physical map comparison discovered that the FAD2 gene and FAE1 gene located on the QTL qA05C18:3 and qA08C22:1confidence interval, respectively. 【Conclusion】 The oleic acid, linoleic acid and erucic acid content QTL was mapped with a new set of rapeseed 60K chip array, and the major QTL on A08 and C03 for erucic acid content is also responsible for oleic acid content. The mapping results are helpful to the fatty acid composition QTL comparison in different populations based on the same rapeseed 60K chip array.

【Objective】 This study aimed to explore the characteristic of nitrogen metabolic in diallel cross generation of different maize genotypes after silking.【Method】Selection high nitrogen use efficiency hybrid maize plays an important role in high yield and high efficiency production of maize A full diallel cross was made with three stay-green inbred lines (Q319, CZ01, VA91) and three non-stay-green inbred lines (BM, B73, MO17) in field. The physiological traits of nitrogen metabolic in diallel cross generation were analyzed.【Result】Content of nitrogen in leaf graphed as inverted “V”-shapes after silking stage in different maize genotypes. The peak of nitrogen content in leaf occurred in filling stage. Content of nitrogen in stem-sheaf and sheath reduced with the extension of the growth duration. Content of nitrogen in leaf, stem-sheaf and sheath of stay-green (SG) inbred lines were much higher than those in non-green-stay (NSG) ones. Nitrogen accumulation before silking stage (NABS), nitrogen accumulation after silking stage (NAAS) and total nitrogen accumulation (TNA) in SG self cross were 20.00%, 82.30% and 45.23%, respectively, higher than those in NSG ones. However nitrogen translocation, nitrogen translocation rate and nitrogen harvest index in SG were 51.28%, 66.59% and 14.32% , which was significantly lower than those in NSG ones. TNA of SG was about 50% and TNA of NSG was only 40% after silking stage. Crude protein content in leaf and stem-sheaf of SG were 108.23% and 33.63%, higher than those in NSG ones. The straws were still higher valuable after harvesting. Some key enzymes to nitrogen metabolic activity such as nitrate reductase (NR) activity, glutamate synthetase (GOGAT) and glutamine synthetase (GS) in leaf of stay-green type were significantly higher than those in non-green-stay types. NR activity in leaf increased firstly and the peak occurred in filling stage after silking stage in different maize genotypes. GS activity, GOGAT activity and soluble protein content in leaf increased firstly, and the peak occurred 15 days after silking. Correlation analysis between stay-green degree, content of nitrogen, soluble protein content, GS activity, GOGAT activity and NR activity indicated that there existed significantly or highly significant positive correlations. Activities of those key enzymes on nitrogen metabolism were significantly important for nitrogen assimilation. 【Conclusion】 High nitrogen metabolism level is one of the nutrition physiological basis which makes leaves keep green for long time. Compared with the non-stay-green types, stay-green genotypes behave higher activities of key enzymes to nitrogen metabolic and higher nitrogen asorption and assimilation at late growth stage. In conclusion, character of stay-green could be used as an important agronomic trait in selecting high nitrogen efficient genotypes and inbred line.

【Objective】A field experiment was conducted to clarify the regulatory effects of potassium application time on starch accumulation and its physiological mechanism, starch granule distribution of storage roots of sweetpotato variety Jixu 23. 【Method】Jixu 23 (cultivar for starch use) was planted at Tai’an experimental site in Shandong Province, with randomized block design in 2008-2009 growing season. There were four treatments in all, including the control (non-potassium fertilizer, T1), potassium applied as basal fertilizer (T2), as top cover fertilizer (T3) and as top-growth peak fertilizer (T4). Samples were obtained by using periodic sampling during storage roots development. The content of starch, amylose and amylopectin of storage root were determined at different growth stages. Meanwhile the medium diameter of starch grain was determined and the volume fraction of different diameters starch granules (＞19.76 μm, 3.359-19.76 μm and ＜3.359 μm) was calculated. The activities of enzymes (SS, ADPGPPase, UDPGPPase, GBSS and SSS) related to starch synthesis were also studied. The effects of potassium application time on starch and its components, starch granule distribution and activities of enzymes related to starch synthesis in storage roots at different growth stages were analyzed.【Result】Compared with the control (T1), dry matter accumulation and starch yield of storage roots in treatments with potassium applied increased, and the increase of starch yield of T2, T3 and T4 treatments was 24.90%, 16.57%, and 9.24%, respectively. And the increase decreased when the application time delayed. The amylose content of storage root in treatments of topdressing (T3, T4) increased while amylopectin content was decreased significantly. The volume fraction of large starch granules in treatment with potassium applied as basal fertilizer (T2) was larger, and the volume fraction of medium and small starch granules in treatment with potassium applied as top-growth peak fertilizer (T4) was larger than that in the other treatments. Potassium application increased the activities of SS, ADPGPPase and UDPGPPase, and enhanced the supply capability of precursors for the synthesis of starch. And the longest action time of potassium on SS and ADPGPPase activities was 130 days, and potassium applied as basal fertilizer (T2) worked effectively mainly at early and middle stages of storage roots development, while treatments of topdressing (T3, T4) worked effectively mainly at middle and late stages of storage roots development. For UDPGPPase activity, the earlier the potassium applied, the longer action time was got. Potassium applied as topdressing (T3, T4) increased SSS activity but reduced GBSS activity of storage roots during the later expanding period. There was a significant positive correlation between total starch content and SS and ADPGPPase activities, and the significant positive correlation was also found between amylose content and GBSS and SS activities. And amylopectin content had a significant positive correlation with ADPGPPase and SSS activities.【Conclusion】Potassium applied as basal fertilizer (T2) and top cover fertilizer (T3) increased dry matter accumulation and starch yield of storage root significantly. Potassium applied as topdressing increased amylopectin content but decreased amylase content significantly. Treatments with potassium applied as basal fertilizer got a higher volume fraction of large starch granules, while treatments with potassium applied as top-growth peak fertilizer got a higher volume fraction of medium and small starch granules. The significant effects of potassium application time on amylase content, amylopectin content and starch yield were closely related with the changes of SS, ADPGPPase, UDPGPPase, GBSS and SSS activities.

【Objective】The objective of this study is to identify the pathogen causing crown rot on strawberry in Wuhan, and to provide a basis for disease prevention and resistance breeding.【Method】Diseased samples from Wuhan, Hubei Province in central China were collected, pieces of infected tissue from the crowns were surface disinfested and plated on acidified potato dextrose agar (PDA). The growing edges of any fungal hyphae developing from the tissues were then transferred aseptically to another PDA plate. Isolates were characterized by colony morphology, conidial characteristics and mycelia growth rate under 18℃ in dark. One-way analyses of variance (ANOVA) were conducted to determine the significance of differences in growth rates of different isolates. According to the morphological characteristics, isolates Zhd-3, Zhd-4-1 and Zhd-5 were selected as representatives for next analysis. Pathogenicity test was carried out by inoculating conidia of the three isolates to crown of strawberry with toothpick, respectively. Conidia produced on petioles were observed by optical microscope and scanning electron microscopy. The length and width of 20 conidia of each isolate were measured. Partial actin (ACT), beta-tubulin (TUB2) and calmodulin (CAL) genes of Zhd-3, Zhd-4-1 and Zhd-5 were amplified and sequenced. The ACT, TUB2 and CAL sequences of Zhd-3, Zhd-4-1, Zhd-5, along with the 23 different representative isolates in C. gloeosporioides species complex were used for multigene loci analyses with Mega 4.1 and Neighbour-Joining method. 【Result】Fifteen isolates were morphologically identified as Colletotrichum spp.. Colony morphology of these isolates on the above of PDA plate appeared similar but significantly different on the back. The 15 isolates were divided into three groups based on the colony morphology, and isolates Zhd-3, Zhd-4-1 and Zhd-5 belonged to each group, respectively, were selected for follow-up tests and analysis. The pathogenicity tests showed that the symptoms identical to those observed in field were produced on all inoculated plants. Control plants remained symptomless. The same pathogen was reisolated from the diseased tissues. Biological tests showed that mycelial growth rates of Zhd-3, Zhd-4-1 and Zhd-5 were 0.82, 0.68 and 0.88 cm?d-1, respectively. The average size of 20 conidia was 11 μm × 3.8 μm. The biological characteristics indicated these isolates were C. gloeosporioides species complex. The result of multigene loci analysis based on ACT, TUB2 and CAL indicated that isolates Zhd-3, Zhd-4-1 and Zhd-5 were divided into one group with C. siamense strains ICMP12567, ICMP17795, ICMP18121, CBS113199 and CBS112983.【Conclusion】The results of biological characteristics and multigene loci analysis based on the sequences of TUB2, ACT and CAL are concluded that the crown rot disease of strawberry occurred in Wuhan is caused by C. siamense.

【Objective】The effects of constant temperatures on the life table parameters of western flower thrips (Frankliniella occidentalis) were well established, and the results were used as a means of projecting the growth of populations under natural condition. F. occidentalis in the wild, however, are not exposed to constant temperatures as they are faced with temperature variation on a daily basis. In this study, life tables of F. occidentalis under constant temperature (CT) and daily fluctuating temperatures (DFT) were compared, and the accuracy of population occurrence dynamics in natural condition based on data from CT were explored. 【Method】Developmental, survival, and fecundity rate data were analyzed using the age-stage, two-sex life table of F. occidentalis reared on bean pod were constructed under constant temperature (CT, 26℃) or daily fluctuating temperatures with same mean (DFT, 20-32℃). As the hatch rate of eggs varies with maternal age, the age-specific fecundity was calculated based on the numbers of hatched eggs to reveal the biological characteristics of F. occidentalis accurately. Means and standard errors of population growth parameters were calculated using the bootstrap method. The Mann-Whitney test (U test) was used to evaluate the differences in the population parameters, development times, and fecundities of F. occidentalis reared under CT and DFT.【Result】There were significant differences in pre-adult developmental time, total preoviposition period, pre-adult survival and life table parameters between F. occidentalis reared under CT and DFT (P＜0.05). However, no significant difference was found in adult longevity and fecundity between F. occidentalis reared under CT and DFT. The pre-adult developmental duration of F. occidentalis reared under CT (11.86 d) was significantly longer than those under DFT (11.36 d). The total preoviposition period of F. occidentalis reared under CT (12.50 d) was significantly longer than those under DFT (11.37 d). The pre-adult survival of F. occidentalis reared under CT (0.48) was significantly higher than those under DFT (0.44). The numbers of eggs laid per female were 35.38 and 34.74 for F. occidentalis reared under CT and DFT, respectively. The intrinsic rate of increase (r), finite rate (λ) and net reproductive rate (R0) of F. occidentalis reared under CT were 0.121 d-1, 1.129 d-1 and 7.538, respectively, while those reared under DFT were 0.127 d-1, 1.135 d-1 and 8.831, respectively. The population growth of F. occidentalis reared under DFT was quicker than those reared under CT. 【Conclusion】The results indicated that by using CT, one could under- or over-estimate the values of numerous life-history traits compared to more natural DFT conditions. This inaccuracy may in turn reduce the accuracy of population surveillance and downstream applications for the timing of integrated pest management.

【Objective】The objective of this study is to analyze the influence of spray method on the deposit and distribution of pesticide within the rice population, and to put forward reasonable spay technology and improve the utilization ratio of pesticide in rice field. 【Method】The solution of ponceau-G was sprayed by manual sprayer with a hollow cone nozzle upon rice plant and mist sprayer with pneumatic nozzle down 45° tilt angle at rice tillering, booting and flowering stages. The ponceau-G deposited on the microslides, which respectively showed the obverse and reverse of rice leaves and the vertical stem of plant, were collected and measured.【Result】There was a significant difference among the deposition amounts of penceau-G sprayed by different methods at different sites within the field rice population at different growing stages. The deposit rate of ponceau-G was manifested with a distribution trend of upper layer＞middle layer＞lower layer. At the rice tillering stage, manual sprayer was applied and the deposition of ponceau-G on 13.3% of microslides was close to the average value, the deposition of ponceau-G 52.9% of microslides was lower than the average value, and 33.75% was higher than the average value. At the rice tillering stage, mist sprayer with pneumatic nozzle down 45° tilt angle was applied and the deposit rate of ponceau-G on 20.4% of microslides was close to the average value, 45.4% was lower than the average value, and 34.2% was higher than the average value. At the booting and flowering stages, manual sprayer was applied and 33.3% and 28.1% of microslides did not have any deposition of ponceau-G. When mist sprayer with pneumatic nozzle down 45° tilt angle was applied, there was 13.9% and 5.0% of microslides did not have any ponceau-G deposition. The deposition of only 6.4% and 11.7% of microslides was close to the average value at booting stage for manual sprayer and mist sprayer. At the flowering stage, the proportion was 7.2% and 17.2%, respectively. For manual sprayer, ponceau-G mainly deposited on the microslides expressing obverse of leaves and the amounts were, respectively, 66.3%, 85.1% and 84. 9% of total deposits on microslides in three rice stages, among which the amount of ponceau-G on the microslides from upper layer of plant were, respectively, 38.7%, 42.2% and 45.6% of total deposition. There were litter deposition on the microslides expressing the reverse of rice leaves and the vertical stem of plant, especially on some microslides put in the base of rice plant, the amount of ponceau-G was zero at rice booting and flowering stages. For mist sprayer, the deposition on the microslides showed the reverse of rice leaves and the vertical stem of plant was more than that of manual sprayer. However, the ponceau-G on the microslides showing obverse of leaves were still 50.5%, 50.6% and 53.1% of total deposits, among which the ponceau-G on the microslides from upper layer of plant was, respectively, 32.3%, 27.9% and 31.5% of total deposition.【Conclusion】If the average of total deposition on each microslides was the pesticide dose controlling pest effectively, there were shortage and waste of dose within rice population at the same time. The fog drip of manual sprayer spraying mainly deposited on the obverse of rice leaves of plants’ upper leaves. The deposition rate accounts for above 1/3 of the total deposition amount, while the deposition rate was few on the reverse of leaves and the vertical stem, especially on the plants’ base leaves and vertical stem, on which the deposition rate was nearly zero. When mist sprayer with pneumatic nozzle down 45° tilt angle was applied, the deposition rate of fog drip on the obverse of rice leaves of plants’ upper leaves was more than 1/4 of the total deposition amount. However, when the application volume was less than that of manual sprayer conducting coping spraying, each layer of the reverse of leaves and the vertical stem was demonstrated a significantly higher deposition rate than manual sprayer. It illustrates that spraying with the mist sprayer with pneumatic nozzle down 45° tilt angle, pesticide droplets could be delivered to middle and lower layer and deposited on rice stem and reverse side of leaves.

【Objective】Due to the widespread use of glyphosate in agricultural production, glyphosate has become the preferred object on transgenic crops resistant to herbicides. Finding glyphosate resistance genes is a primary issue in the study of transgenic crops resistant to glyphosate. There are various kinds of microorganisms in nature with rich genetic resources, so this study intends to screen and identify high glyphosate-resistance bacterium strains from field soil samples of Guangdong area, then clone the target gene of glyphosate from the strain and test its glyphosate-resistance level, in order to obtain high glyphosate resistance gene resources for research of glyphosate resistant transgenic crops.【Method】The gradient dilution glyphosate selected culture method in the isolation of high glyphosate-resistance bacterium from field soil samples of Guangdong area was applied. Identification of the strain species according to the microscopic observation, gram staining and 16 S rDNA sequence analysis results. Using RT-PCR method to clone the strain’s target gene of glyphosate, and analyse the basic characteristics of aroAS001 sequences by sequence alignment and phylogenetic tree. The aroAS001 variant performed site directed mutagenesis by overlapping PCR method to obtain aroAS001-mut gene fragments. The aroAS001 and aroAS001-mut fragments were transferred into defective Escherichia coli DH5α/△aroA strains, respectively, to detect the resistant levels of glyphosate. 【Result】 A high glyphosate-resistance bacterium strain was isolated, and it was identified as Klebsiella pneumoniae by morphological and molecular biology methods, named kpS001 strain. The target gene of glyphosate from kpS001 strain named aroAS001 was cloned. Sequence analysis showed that the amino acid sequence of the gene encoded had a typical Class I EPSPS features, and there was a single amino acid different from with another K. pneumoniae strain’s aroA. After obtained the aroAS001-mut fragments by overlap PCR, aroAS001 and aroAS001-mut fragments were transferred into the defective E. coli DH5α/△aroA strains, respectively, to detect the resistant levels of glyphosate. Compared with the control strains, the recombinant strains containing aroAS001 and aroAS001-mut were able to grow normally in the culture medium containing less than 200 mmol?L-1 glyphosate, however, as glyphosate concentration increasing, the growth state of the recombinat strains gradually suppressed, while the concentration of glyphosate increased to 350 mmol?L-1, the growth was completely suppressed. 【Conclusion】The K. pneumoniae S001 is a high glyphosate-resistance bacterium, the target gene of glyphosate aroAS001 belongs to the Class I EPSPS and shows a significant glyphosate-resistance characteristics, it could be used as potential gene materials in transgenic glyphosate-resistant crop studies.

【Objective】The potassium (K) natural productivity from soil, K fertilizer use efficiency and apparent soil K balance were investigated to provide a theoretical basis for the sustainable management of K element in farmland. 【Method】The three fertilizer treatments used were control, NP and NPK, representing various combinations of N, P and K fertilizer applications. Long-term (over 20 years) experiments with barley-rice crop rotation were conducted to investigate the response of crop aboveground biomass, yield, the K content in grain and straw to different application treatments. 【Result】The results indicated that the grain yields on average were 8.24 and 9.22 t?hm-2 per year in barley-single crop rice and barley-early rice-late rice rotation cropping system without application fertilization for 20 years, which accounted for 75.6% and 71.9% of that in NPK treatments in the corresponding rotation system. The contribution of application fertilizers increased crop grain yields by 24.4% to 28.1%, and the contribution of K fertilizers increased crop grain yields by 11.8% to 14.0% in barley-single crop rice and barley-early rice-late rice rotation cropping system, respectively. The contribution of K fertilizers increased grain yields for barley (17.9%) was 36.1% higher than that for rice. The annual K uptakes from CK treatments accounted for 59.1% and 58.3% of that in NPK treatments. The K in straws accounted for 81.7% and 74.2% to 87.6% of total K uptake for barley and rice, respectively. The indigenous K supplying capacity in barley season was lower than that in rice season. The annual indigenous K supplying capacity for barley on average was 69.8%, which were 15.5%, 19.7% and 19.4% lower than that for early rice, late rice and single crop rice, respectively. The indigenous K supplying capacity in barley season decreased significantly over cultivation time at an average rate of 1.6% per year. But there was no significant change of the indigenous K supplying capacity in time during rice cropping. The indigenous K supplying capacity in rice season still was more than 80% after 20 years cropping. The internal K use efficiency was decreased because of K application. But there was no significant change of internal K use efficiency among treatments in time. Under balanced fertilization with fixed N, P and K inputs, the internal use efficiencies for barley, early rice, late rice and single-crop rice were 42.4, 44.7, 41.3 and 44.8 kg•kg-1, respectively. The partial factor productivity in NPK treatment was 83.5, 79.7, 71.6 and 69.7 kg•kg-1, respectively. The K use efficiency in barley season was higher than that in rice season. The apparent K recovery efficiency was 29.5% and 41.5% in barley-single crop rice and barley-early rice-late rice rotation system, respectively. Crop continuous uptake caused a net K lost of 148.2 and 182.7 kg•hm-2 per year in the soils without application of fertilizer in barley-single crop rice and barley-early rice-late rice rotation cropping system, respectively. Moreover, the application of nitrogen and phosphate fertilizer accelerated soil K depletion, the extra K amount of 57.8 and 65.0 kg•hm-2 was taken up by crop. Although K deficiency in soil could be relieved by K application, a net K lost of 146.5 and 85.5 kg•hm-2 per year in soils received K fertilizer from barley-single crop rice and barley-early rice-late rice rotation cropping system. 【Conclusion】There was a very strong capacity to provide K element from soils during rice growing season. But the K fertilizer use efficiency in barley season was higher than that in rice season. There was a still net K loss from soil with K application rate of 187.5 kg K2O•hm 2 per year. So a great attention should be paid to the apparent soil K balance.

【Objective】The objective of this study was to investigate the spatial distribution characteristics and quantitative relations of total salt content (TS), pH value and ion components so as to evaluate the relationship and correlation between them and their influences on types and distribution of saline-alkali land in Gansu Yellow River Irrigation District, Northwest China. 【Method】Take the typical irrigation area of Xingdian and Jingdian as research soil sampling area with sampling sites fully covered the newly reclaimed saline-alkali wasteland in the upper reaches, the mildly salinization of cultivated land in the middle reaches, the moderately severe salinization of cultivated land in the lower reaches and the salinization abandoned cropland in the whole irrigation area. And given more consideration to the differences between the types (sulfate type, chloride-sulfate type, sulfate-chloride type) and the degrees (mild, moderate, severe) of soil salinization when sampling. The conventional statistics method was used to evaluate the soil profile distribution of TS, pH and salt ion. The canonical correspondence analysis method was used to evaluate the quantitative relationships of TS, pH and the contents of salt ion, and the correlation coefficients, eigenvalues, cumulative percentage variance of TS, pH, salt ions between axis 1 and axis 2 of CCA in various soil layers. 【Result】 The irrigated soils of Gansu Yellow River Irrigation District are mild to severe salted, the salt content of 0-40 cm soil layer decreased with soil depth increase while the salt content of 40-80 cm soil layer increased with soil depth increases. The cation and anion in soil profile are Na+ and SO42 respectively -with pH value vary between 7.93 and 9.98. K+ and HCO3-, Ca2+ and Mg2+ are well linked in all soil layers, the correlation between Na+ and Cl- are enhanced with increment of soil depth, SO42- and Na+ are correlated negatively, the correlation between K+ and Na+ as well as SO42- , Cl- and HCO3 are not stronger in all soil layer-. At 0-100 cm soil profile, the spatial distribution of pH is mainly controlled by HCO3-, TS is mainly controlled by Na+, Cl- and SO42-, respectively. Different soil layers have diverse characteristics of soil salinization from that of the whole soil profile, the spatial distribution of pH and TS is mainly controlled by SO42- in 0-20 cm soil layer, by Cl- and Na+ in 20-40 cm soil layer, by Mg2+ in 60-80 cm soil layer, by Mg2+ and Cl- in 80-100 cm soil layer, by Ca2+, Mg2+, SO42- and HCO3- together in 40-60 cm soil layer individually. Axis1 and axis 2 of CCA reflects the changes of TS and pH value, respectively. The eigenvalues and cumulative percentage vary from 0.53 to 0.70 and 38.7% to 57.7% of axis 1, respectively, while that vary from 0.36 to 0.64 and 30.6% to 44.7% of axis 2 correspondingly. 【Conclusion】There are two processes of surface salt accumulation and bottom salt accumulation profiles in the saline cultivated land of Gansu Yellow River irrigation District. Newly reclaimed saline-alkali wasteland in the upper reaches and mild salinization of cultivated land in the middle reaches are given priority to surface accumulation profiles, and mild salinization of cultivated land in the middle reaches and moderately severe salinization of cultivated land in the lower reaches are given priority to bottom accumulation profiles. The salt content of 60-80 cm argillaceous cemented sand layer is as high as 93.7 g•kg-1, which has an important influence on the migration of water and salt and the distribution of salt ions, and is one of the main causes to lead soil to secondary salinization in the irrigation area. TS, Na+ and SO42- are the main factors that determine the type and distribution of saline-alkali land in Gansu Yellow River Irrigation District. Meanwhile, the canonical correspondence analysis method can intuitively give the relationship among various salt ions and between TS, pH and salt ion in two-dimensional ordination diagram of CCA by the length of the arrow, the quadrant, the angle between arrows, the distance between arrows and the triangle symbol, etc. And it also can determine the main controlling factors that influence the characters of different layers and the main factors that affect the type and distribution of saline-alkali land by using correlation coefficients, eigenvalues, cumulative percentage between TS, pH, salt ions and axis1/axis2 of CCA in various soil layers, so that, it is considered to be the more efficient scientific method for evaluation of the characteristics and improvement of saline-alkali land in Yellow River Irrigation Area.

【Objective】In this study, the method of cabbage DNA fingerprint was drawn, and fifty cabbage representative varieties from China were fingerprinted with EST-SSR primers to provide reference for variety distinctness, authenticity, and purity identification. 【Method】First, EST-SSR primers were screened by using the technologies of 6% denaturing polyacrylamide gel electrophoresis and six cabbage varieties that come from different ecogeography. The length of amplified fragments was detected on DNA Analyzer platform using four fluorescent-labels (TAMRA, HEX, ROX and 6-FAM) in 5′end of forward primer, and then defined the reference variety for every alleles. Total twenty core primers were used to establish fifty cabbage varieties SSR fingerprinting, and for ‘Zhonggan 21’ variety identification. 【Result】 Six cabbage varieties of different resources were used to screen 978 EST-SSR primers, out of 128 polymorphic primers were obtained according to the PCR bands stability, high polymorphism information content (PIC), easy discrimination of different alleles and even distribution of molecular markers on each chromosome, and 20 pairs of primers were selected to detect a total of 58 alleles at 20 loci, with 2.22 loci per chromosome and 2.9 alleles per locus on average. The PIC values varied among the primers ranging from 0.34 to 0.76. The length of amplified fragment varied in the range of 143-296 bp. The maximum number of alleles for each primer pairs of BoE607 and BoE723 was five. Fingerprinting database of 50 cabbage representative varieties from China was established with 20 pairs of core primers. The authenticity of ‘Zhonggan 21’ was identified by artificial simulated population and these results were identical with that made from field investigation.【Conclusion】Twenty pairs of core primers were selected and used to establish DNA fingerprint database of 50 cabbage representative varieties from China, and the authenticity of ‘Zhonggan 21’ was identified by artificial simulated population and the accuracy was 100%.

【Objective】Cucumber (Cucumis sativus L.) is an important fruit vegetables. Fruit quality is always getting more attention in cucumber breeding program. Fruit quality includes inner quality and exterior quality, and fruit exterior quality of cucumber has important influences on its commodification. Spine color is one of the important fruit quality traits in cucumber. The clarification of the inheritance and identification of molecular markers for the fruit spine color gene will provide a theoretical basis for breeding of fruit quality and lay a foundation for fine mapping and gene cloning. 【Method】 Cucumber inbred lines GY14 with white fruit spines and NC76 with black fruit spines were used as the experiment materials for genetic analysis and gene mapping for black fruit spine in this study. Bulked segregation analysis (BSA) was performed in the F2 population using 2112 SSR markers. The sequence and re-sequencing information of 9930 and 100 core germplasms were used to develop new SSR and Indel markers in the primary mapping region of the black spine color gene (B). JoinMap 4.0 and MapInspect software were employed to construct a linkage map for the B gene with SSR markers. Bio-informatics was adopted to predict candidate genes in the genomic region harboring the B gene. A set of 156 recombinant inbred lines (RILs) were used to test the veracity for marker-assisted selection (MAS) of flanking molecular markers linked to the B gene. 【Result】Genetic analysis showed that the trait of black fruit spine in NC76 was qualitative, and a single dominant nuclear gene (B) controlled this trait. Black was dominant to white. In the primary genetic mapping of the B gene, eight SSR markers were screened to be linked with the black fruit spine color locus. The B gene was mapped on the chromosome 4 (Chr.4) of cucumber. The closest linked marker SSR22231 was 10.8 cM away from B. A total of 212 pairs of new SSR primer and 25 pairs of Indel primer were developed based on the sequence information in the primary mapping region of B. Fourteen SSR markers and one Indel marker were identified to be linked with the B gene after analysis of the F2 mapping population using these new developed molecular markers. The two closest flanking markers SSRB-181 and SSRB-130 were 2.0 and 1.6 cM away from B, respectively. The physical distance between SSRB-181 and SSRB-130 was 422.1Kb containing 60 predicted genes. Csa4G003095 and Csa4G001690 were the most possible candidate genes for the black fruit spine trait. The probability of the two flanking SSR markers to predict the spine color for marker-assisted selection (MAS) breeding was 96.8% and 96.2%, respectively. And the accuracy rate for SSRB-181 to predict the black spine color in MAS breeding was 100%.【Conclusion】The black fruit spine trait of NC76 is controlled by one dominant nuclear gene (B). It is located on the Chr.4 of cucumber delimited in a physical distance of 422.1 Kb. The two closest flanking markers SSRB-181 and SSRB-130 could be used in the MAS breeding program. The results in this study will be of great benefit to fine mapping and gene cloning for the B gene and lay a good foundation for cucumber MAS breeding.

【Objective】 The objectives of this study were to determine the structures of CHS genes in Camellia sinensis (CsCHS) by obtaining gDNA sequences of these genes, and to analyze single nucleotide polymorphism. Besides, association analysis was also carried out in order to find potential SNP sites in CsCHS which would influence the polyphenol content in tea plant.【Method】 Based on CsCHS sequences uploaded to NCBI, specific primers were designed using primer 3.0 software. Genomic DNA and cDNA of tea leaves were used as templates in polymerase chain reaction (PCR) to obtain gDNA and cDNA sequences of CsCHS1, CsCHS2 and CsCHS3, respectively. Gene structures were determined through blasting gDNA and cDNA sequences. The putative amino acid sequences were analyzed by bioinformatics softwares, such as Compute pI/Mw, SOPMA, and so on. Single nucleotide polymorphism of CsCHS were analyzed in 57 cultivars with great variation polyphenol contents. In order to obtain the coding region of CsCHS genes, PCR reactions with specific primers were carried out by using cDNA of individual tea cultivars as template. TASSEL software was introduced in association analysis.【Result】 cDNA sequences of CsCHS1, CsCHS2 and CsCHS3 were 1 277 bp, 1 320 bp and 1 242 bp, respectively. And an open reading frame (ORF) of 1 170 bp was found in each CsCHS gene. gDNA sequences of CsCHS1, CsCHS2, and CsCHS3 were 1 600 bp, 1 330 bp and 1 607 bp, respectively. By comparing gDNA and cDNA sequences of each CsCHS gene, combined with GT-AG rule, it was determined that CsCHS1 and CsCHS2 have two exons and one intron, respectively. And the intron in CsCHS1 is 323 bp, compared with 356 bp in CsCHS3. While no interruption region was found in CsCHS2, this might prove there is no intron in CsCHS2. Deduced amino acid sequences analysis suggests that identity of amino acid sequences are 92.6%-95.4%. All of the conserved amino acids found in CHS protein subfamily were also found in these deduced sequences. Furthermore, bioinformatics analysis showed highly similarities among CsCHS1, CsCHS2 and CsCHS3 protein structures. There are 71 SNP sites in CsCHS1’s coding region, SNP frequency was 1SNP/16.48 bp, and no Indel was found. A total of 55 SNP sites were found in CsCHS2’s coding region, suggesting one SNP in every 21.27 bp. And the nucleotide diversity (π) in CsCHS1 (0.01088) was significantly higher than that of CsCHS2 (0.00530). By correlation analysis, two SNP sites were positioned thought to be related to polyphenol content in tea plant in CsCHS1 and 4 in CsCHS2. No further analysis referring to CsCHS3 was carried out due to low success rate of PCR reaction.【Conclusion】 Both CsCHS1 and CsCHS3 were determined to be conserved CHS genes, and bioinformatics analysis of protein structures results show that CsCHS1, CsCHS2 and CsCHS3 are similar. CsCHS1 and CsCHS3 are active, there should be hot spots of mutation in their coding regions.

【Objective】Pre-slaughter fasting is defined as duration of food and water withdrawal during loading, unloading, transport and control during pre-slaughter lairage period. Impropriety of pre-slaughter handling decreases carcass yield, affects eating quality and technical characteristics of meat. The objective of this study was to investigate the effects of different pre-slaughter fasting time on quality of mutton and determine how long fasting time is suitable for production of high quality mutton.【Method】Thirty six-month old sheep were divided into 3 groups fasting for 0 h(control group), 12 h and 24 h. The eating quality (pH value, meat colour, shear force and water holding capacity), sensory quality (odour, tenderness, juiciness and overall liking), hygiene quality (aerobic plate count and coliforms), glycogen concentration, sarcomere length and protein degradation during post-mortem were analyzed.【Result】Fasting for 24 h resulted in higher pH value than 12 h and control group at 0 h, 45 min and 4 h post-mortem. As aging time extended, pH value at 24 h and 48 h post-mortem had no significant difference (P＜0.05). The group fasting for 24 h had lower cooking loss than 12 h and control group (P＜0.05). There was no significant difference among three groups in L*, a*, b*,ΔE, drip loss, shear force and sensory characteristics of odour, tenderness, juiciness and overall liking of the mutton (P＞0.05). Aerobic plate count and coliforms of the mutton from different fasting time groups were all under the range of national standard regulations at 24 h post-mortem. Aerobic plate count of mutton had no significant difference among three treatments. Coliforms of the mutton were decreased as the fasting time increased. Fasting treatments caused the decrease of enumeration of coliforms compared to control group. There was no significant difference in glycogen content of mutton at 0 h, 45 min, 4 h, 8 h, 12 h, 24 h and 48 h post-mortem among three groups. The degree of glycolysis increased but the glycogen content decreased as the extension of aging time. With the increase of pre-slaughter fasting time, sarcomere length increased. Fasting for 24 h resulted in larger sarcomere length than 12 h and fasting for 12 h was higher than 0 h (P＜0.05). The degree of myofibrillar protein degradation in different fasting treatments was consistent at 0 h post-mortem. There was no apparent bands appeared or disappeared. All groups had 27 kD fragment appeared at 24 h post-mortem. The appearance of the 27 kD was darker in fasting for 12 h and 24 h treatments than the control. It was shown that fasting for 24 h and 12 h resulted in higher protein degradation degree than the control group.【Conclusion】Fasting for 12 and 24 h increased the hygiene quality and accelerated meat myofibrillar protein degradation. Fasting for 24 h decreased cooking loss. Fasting for 12 and 24 h had no significant effects on pH value, glycogen content and other quality characteristics of mutton. Compared with the control group, fasting for 12 and 24 h had positive effects on mutton hygiene quality but no effect on eating quality and sensory quality.

【Objective】Skeletal satellite cells are activated by some specific stresses such as development and trauma, and differentiate and form myotubes to participate in the development or repair of skeletal muscle. FoxO1 negatively controls the genesis of skeletal muscle, but the molecular mechanisms by which FoxO1 funcions in the differentiation of satellite cells have not been reported so far. This experiment was conducted to explore the effects of FoxO1 on porcine skeletal muscle satellite cells differentiation, aiming to provide new theoretical reference for further research. 【Method】Extensor digitorum longus of 1 to 3-day-old piglets were used to isolate the skeletal muscle satellite cells and the cells were observed and pictures were taken by inverted microscope on day 2, day 4 and day 6, respectively. The cells were stained by immunofluorescence staining and DAPI nuclear staining on day 8 of differentiation, and observed under a fluorescence microscope. Meanwhile, the medium was replaced with differentiation medium containing 50 nmol•L-1 wortmannin (wortmannin, WM) differentiation medium when the cells density reached 70% -80% confluence, the cells were collected on day 0, day 4, and day 8, respectively. Total RNA and total protein were extracted, and Real-time qPCR and Western blotting were performed to measure the alterations in the expression of FOXO1 and myogenic differentiation marker genes caused by WM supplement. 【Result】Porcine skeletal muscle satellite cells became adherent to the dish bottem, spindle-shaped on day 2. Cell number increased on day 4 and some cells started to fuse. On day 6, cell started to grow with directivity. On day 8, cell further fused to form myotubes, further fused to form myotubes. There was no significant difference in mRNA expression level of FoxO1 between WM treatment group and the control (P＞0.05), unphosphorylated FoxO1 increased significantly (P＜0.05) with WM treatment, whereas phosphorylation level of FoxO1 dropped drastically (P＜0.05). Although on day 8 the cells displayed an alveolate morphology after treated with WM, they failed to show directional growth and formation of myotubes. Moreover, Western blotting results demonstrated that WM decreased the protein level of MyoD (early myogenic marker), MyoG (middle-stage marker), and MyHC (late marker) significantly.【Conclusion】Results of the study suggest that inhibition of PI3K signaling pathway by WM blocks results in FoxO1 phosphorylation, suppression of porcine skeletal muscle satellite cell differentiation, delay of the formation of myotubes, and down-regulation of myogenic differentiation marker genes, such as MyoD, MyoG, and MyHC. Take together, blockade of PI3K signaling pathway suppresses porcine skeletal muscle satellite cell differentiation through the activation of FoxO1.

【Objective】Growth differentiation factor-10 (GDF-10) is a member of the TGF-beta super family and the bone morphogenetic protein family (BMP). It is originally identified by PCR using degenerate oligonucleotides based on the BMP-3 sequence. Studies suggested that the GDF-10 gene plays a key role in skeletal morphogenesis. The objective of this study was to identify the polymorphisms of GDF-10 gene and their correlation with production traits was investigated for the purpose of the breeding of Gannan yak. 【Method】 Thirty samples were randomly selected from genomic DNA of 298 Gannan yak to construct DNA pool. Nine primers were designed to amplify the exons 1, 2 and 3 of GDF-10 gene. Effective PCR production was purified by DNA gel extraction kit and direct sequencing subsequently. BLAST and Chromas were used to screen the SNPs in GDF-10 gene by the wave height of sequencing map. The genotypes were determined by high-resolution melting curre (HRM) and the frequency of alleles was estimated. The association between genotypes and body measurement was evaluated by software SPSS13.0. Three SNPs (T132C, G672T and G1204A) of GDF-10 gene were identified in the population, which were at extrons 1,extron 2, and extron 3, respectively. The PHASE and SHEsis softwares were used to analyze matching chain disequilibrium and haplotype analysis, respectively. The SPSS17.0 software was used for association analysis. 【Result】Three SNPs, 12116 (G/A), 12152 (C/T) and 13041 (T/C) were discovered in exon 3. Through population genetics analysis, the results showed that three locals were at low polymorphic status (PIC＜0.25). The ?2 tests showed that the locus of 13041 (T/C) was not all in the status of Hardy-Weinberg equilibrium (P＜0.05), the other two loci were all in the status of Hardy-Weinberg equilibrium (P＞0.05). Analysis of matching chain disequilibrium and haplotype analysis show that there is a weak chain of balance between the three locals. Analysis of association of polymorphism with body measurement trait at all mutation loci show that there were significant effects on body height, body length, weight and herrt girth (P＜0.05 or P＜0.001) and there is no effects on tube girth (P＞0.05). Seven haplotype combinations were found in 298 Gannan yak, the ATC and ATT have significant effects on body measurement traits and weight. 【Conclusion】 The results suggest that three SNPs and two haplotype combinations might have potential effects on production traits in the above mentioned yak populations and could be used for marker-assisted selection.

【Objective】 Locoweed is one of the poisonous weeds in western China that influences the healthy development of grassland and animal husbandry. Locoweeds mainly belong to Astragalus and Oxytropis genera. As a serious lack of forage in winter season, livestock such as cattle and sheep are forced to feed them. Locoweed can decrease the production performance, toxicity and death of animals, and the direct economic loss of the grassland ecological and animal husbandry is about billions of dollars, which is the most serious poisonous weed. At present, the economic losses of animal death caused by locoweed poisoning continues to soar. Swainsonine is the main toxic component and believed to be the cause of animal locoweed poisoning. Locoweed is widely distributed and is rich in resources, and how to improve the grassland ecology and use of locoweed is becoming the topic of research subject and direction. In recent years, with the study and expansion of locoweed, people have found that swainsonine not only has a good anti-tumor effect, also it can be used as an immunomodulator, anti HIV and anti proliferation inhibitor, anti viral and cell protective agent and other drugs. At present, the domestic research on swainsonine anti-tumor and regulation of immunity is hot, However, no research reports are found in antiviral activity and mechanism of action of swainsonine up to the present. This experiment was designed to investigate the anti-virus mechanism of the swainsonine from alkaloid of Astragalus on bovine viral diarrhea virus (BVDV), and to make clear of the antiviral effect in vitro, it will provide references for the screening of anti BVDV drugs. 【Method】 Using cell culture techniques, and combined with CPE observation method and MTT colorimetric method, different concentrations SW toxicity with bovine kidney primary cells (Madin-Darby Bovine Kidney Cells, MDBK) were detected, the concentration of the drug’s safety and TD50 was determined. They were measured by four administration routes of virus before swainsonine, virus after swainsonine, virus-and-swainsonine after 2 hours, and virus-and-swainsonine after swainsonine, which are the effect of different concentrations of SW against blocking effect on BVDV invasion, replicatting inhibition, directly killing effect and combining effects, respectively. The treatment indexes under different interaction modes were also calculated.【Result】It was found that the TCID50 of BVDV-MDV is 10-4.7. SW was not toxic to MDBK when its concentration was below 0.256 μg•mL-1, and TD50 is 2.512 μg•mL-1. Under the way of virus after SW, IC50 is 2.399 μg•mL-1, TI is 1.05. Under the way of SW after virus, IC50 is 1.018 μg•mL-1, TI is 2.47. Under the way of 2 h after mixed virus with SW, IC50 is 1.205 μg•mL-1, TI is 2.08. Under the way of synthetic action, IC50 is 0.782 μg•mL-1,TI is 3.21. After the TI of different modes of action was compared, it showed that the comprehensive effect on BVDV of SW (65.29%，P＜0.01) and the inhibition effect of SW on the reproduction of BVDV (65.05%，P＜0.01) were better, and SW had a certain direct killing effect on BVDV, but the blocking effect of SW on the invasion of BVDV was poor.【Conclusion】The effect of swainsonine (SW)-anti-BVDV activity in vitro was better, and the authors infered that the antiviral mechanism of SW might be that it could enter into the cells and inhibit the reproduction of BVDV, or it could play a role by killing the uncombined BVDV.

【Objective】 The sugar-nitrogen characteristics of wheat at yield level of 9 000 kg•hm-2 following rice were studied. In addition, the sugar-nitrogen nutrition diagnosis indexes for wheat at yield level of 9 000 kg•hm-2 were suggested. 【Method】 Wheat populations at two different yield levels in rice-wheat rotation, including ＜9 000 kg•hm-2 population (hereinafter as HY) and ≥9 000 kg•hm-2 population (hereinafter as SHY), were established by agronomic management of the medium-gluten wheat, Yangmai 20 (Triticum aestivum L.). From 2010 to 2012, field experiments were conducted as split-plot design at 16 combinations of two applications of nitrogen amount (210 kg•hm-2 and 262.5 kg•hm-2), two nitrogen applied ratios of basis﹕tillering﹕elongation﹕booting (3﹕1﹕3﹕3 and 5﹕1﹕2﹕2), and four nitrogen topdressing stages (flag leaf stage, booting stage, heading stage and anthesis). The dynamics of soluble sugar content, nitrogen content and soluble sugar to nitrogen ratio (hereinafter as C, N and C/N) in plant population of different yield levels were analyzed. And at different growth stages, the relationship between plant C, N and C/N and grain yield were analyzed, respectively. In addition, the sugar-nitrogen nutrition diagnosis indexes for wheat at yield level of 9 000 kg•hm-2 were suggested. 【Result】 As the developmental process goes on, the plants C and C/N had two peak values at the beginning of over-wintering stage and at stage from booting to anthesis, and a low value at green-turning stage. Then, the plants C and C/N showed a gradual decrease after anthesis. The plants N, however, showed a gradual decrease during the growth period - rapid decrease during the period between the beginning of over wintering stage and elongation stage, slow decrease during the period between elongation stage and booting stage, and slow decrease during the period between booting stage and maturity stage. In SHY plants, C and N at booting and N at anthesis were significantly higher than those of HY. But C at maturity was significantly lower than those of HY in 2010-2011, and not-significantly lower than those of HY in 2011-2012. In addition, C/N at maturity was significantly lower than those of HY. Plants C and N at booting stage and N at anthesis were in a positive linear correlation with grain yield. Plants C and N at milk-ripe stage were in a parabolic cure relationship with grain yield. Plants C at maturity were in a negative linear correlation with grain yield. The sugar-nitrogen nutrition diagnosis indexes for the population of about 9 000 kg•hm-2 were suggested. At the stages of booting, anthesis, milk-ripe and maturity, C were 14.56%-16.78%, 14.52%-16.82%, 10.59%-11.23% and 1.62%-1.76%, respectively, N were 1.55%-1.64%, 1.47%-1.57%, 1.28%-1.30% and 1.15%-1.20%, respectively, and C/N were 9.37-10.25, 9.80-10.69, 8.29-8.77 and 1.41-1.48, respectively. 【Conclusion】 Compared with HY, from the beginning of over wintering stage to elongation, C and N in SHY plants were generally high, and C/N was relatively suitable. From booting stage to anthesis, C and N in SHY plants were significantly higher. After anthesis, SHY plants could accumulate more sugar and had a higher conversion rate of sugar to grain yield as compared with HY.

【Objective】The objective of this study is to compare the activities of defense enzymes in different tissues of winter jujube induced by the damage of Apolygus lucorum and artificial needling, and to observe the changes of stress defense activities of enzymes in leaves, buds, flowers and young fruits with different levels of damage by A. Lucorum. 【Method】The experiments were conducted with vulnerable jujube tissues inoculation, artificial simulated needling, counting damage spots after 24 hours, then the damaged tissues were sampled. The activities of SOD, POD and CAT were determined by nitroblue tetrazolium method, guaiacol method and ultraviolet absorbance method, respectively.【Result】The activity of defense enzymes was changed with different damages. The activities of SOD, POD and CAT in leaves induced by the damage of A. lucorum and artificial needling were significantly higher than the leaves without damage, and the activities of SOD and CAT induced by A. lucorum damage were significantly higher than the artificial needling. There was not significant difference among the SOD activities in buds without damage, and those damaged by A. lucorum and artificial needling, while POD activities were significantly increased with the damage of A. lucorum and artificial needling. CAT activity in buds induced by A. lucorum damage was significantly higher than needling ones. At the flowering stage, SOD and CAT activities had no significant difference while POD was significantly increased with the damage stress, and that induced by A. lucorum damage was significantly higher than needling ones. At the young fruiting stage, the POD activities which were induced by A. lucorum damage and artificial needling were significantly higher than those without damage. At the same time, SOD activity was significantly higher in A. lucorum damaged fruit, while CAT activity was statistically indifferent. These results indicated that the defense enzymes in the tissues of winter jujube varied at the damage stress. Stress defense enzyme activities of winter jujube differed to varied damage levels of A. lucorum. With increasing damage, SOD activities in leaves and flowers increased initially and then decreased, while the activities of POD and CAT increased. With the increase of damage level, SOD activity in buds was statistically indifferent, while that of POD and CAT increased significantly. With the increasing damage, SOD activity of young fruits decreased initially, and then increased, however, that of POD increased initially and then decreased. 【Conclusion】The damage of A. lucorum could induce a series of biochemical reactions in host-plants. The activities of SOD, CAT and POD in leaves and the activity of POD in the reproductive organs (buds, flowers and fruits) were much higher under the damage stress. Both physical and chemical damages of A. lucorum induced more defense enzymes than the artificial needling. Defense enzymes in different tissues of winter jujube showed variations with the increasing levels of damage.

【Objective】This study was conducted to investigate the effects of ovariectomy on the meat performance, serum GH (growth hormone) levels and the relative expression levels of GHR (growth hormone receptor) mRNA in longissimus and biceps femoris tissues, liver and perirenal fat tissue, and to reveal the reasons of fattening after ovariectomy in female goats.【Method】Forty young hybrid does Kid of Boer Goats (Boer goat ♂ × Guanzhong dairy goat ♀) with similar body weight at about 5 months old were randomly divided into treatment group (n=20) and control group (n=20), the works of insect repellent and epidemic prevention were implemented a month ago before the beginning of the experiment. The goats in the treatment group were ovariectomized at the beginning of the experinment and the goats were not done in control group. Venous blood samples (10 ml) were collected from each goat to separate serum via jugular vein puncture using clean sterile syringes and needles with no additive at 8:00 in the morning on days 0, 10, 20, 30, 40 and 50 during the experiment. Then the serum was separated from blood samples to determine the levels of the growth hormone (GH) with specific ELISA kits (enzyme-linked immunosorbent assay kit) according to the manufacturer’s instruction. The slaughter rate, lean percentage of carcass, loin eye area and proportion of bone and meat were measured after 10 goats (5 goats of each group) were slaughtered on the 50 th day. At the same time, longissimus and biceps femoris tissues, liver and perirenal fat samples were collected immediately after goats were sacrificed, then three biological replicates were collected at each sample, and the samples were rinsed with sterile saline to remove blood clots and wrapped in aluminum foil with good marks, shock-frozen in liquid nitrogen. The mRNA expression levels of GHR gene in liver, longissimus muscle, biceps femoris and perirenal fat were detected with real-time quantitative PCR (qRT-PCR) and the method of 2-ΔΔCt (the Ct mean value of threshold cycle). 【Result】The serum GH levels of the ovariectomized goats on the 20th day showed an upward trend and were up to 9.50 μg•L-1 which were significantly higher than those of the controls (P＜0.05). On the 50th day, the levels were up to 11.55 μg•L-1 that was 2.21 μg•L-1 higher than those of the control group (9.34 μg•L-1) significantly (P＜0.01). In addition, the goats in treatment group with higher slaughter rate, lean percentage of carcass and loin eye area compared with that in the control group. The slaughter rate in ovariectomized goat was up to 45.62% which was higher than that in the control group (42.41%) and the difference was significant (P＜0.05), but the goats had a lower proportion of bone and meat (P＜0.05) in treatment group. In this study, the expression levels of GHR gene in liver of ovariectomized goats were 3.21 times (P＜0.01) more than that of the control group. Compared with the control group, a very significant increase in the GHR mRNA level was observed in longissimus of ovariectomized goats, and the expression level was the highest in the present study which was 4.87 times higher than that of the control group (P＜0.01). At the same time, a higher expression of GHR mRNA in biceps femoris (3.17 times) was also observed in ovariectomized goats, and the difference was significant compared with the control group (P＜0.05). As to perirenal fat, the expression levels of GHR gene in treatment group was 1.91 times more than that in the control group, and the difference was significant (P＜0.05)【Conclusion】 Results of the study showed that there were significant effects of ovariectomy on slaughter rate, proportion of bone and meat, serum GH levels. Meanwhile, the expression levels of GHR gene in liver, longissimus muscle, biceps femoris and perirenal fat tissues of female goats were also influenced by the ovariectomy. It is indicated that the ovariectomy increased the slaughter rate and decreased the proportion of bone and meat through enhancing the levels of serum GH and expression levels of GHR gene in the target tissues, and improved the meat performance of ovariectomized goats finally.

【Objective】 The sucrose non-fermenting protein kinase (SnRK) is a kind of Ser/Thr protein kinase found widely in plants and participates a variety of transductions in signaling pathway in plants. TaSnRK2.10 is an important SnRK2 member involved in response to various abiotic stresses in wheat (Triticum aestivum L.). The objective of this study is to detect the single nucleotide polymorphism (SNP) of TaSnRK2.10, develop and map its functional markers, analyze the relationship between its haplotypes and phenotypic traits, and provide a basis for the genetic improvement and germplasm enhancement by molecular marker assisted selection in wheat. 【Method】 Thirty hexaploid wheat accessions with high polymorphism and their diploid and tetraploid wild relative species were selected to detect the nucleotide polymorphism in TaSnRK2.10-A gene by sequencing. A set of Chinese Spring nulli-tetrasomic lines and the recombinant inbred lines (RIL) derived from a cross of “Yanzhan 1 × Neixiang 188” were used to map TaSnRK2.10-A on chromosome. Based on the polymorphism in the sequence of TaSnRK2.10-A molecular markers were developed. The relevance between TaSnRK2.10-A haplotypes and phenotypic traits was analyzed using a natural population consisted of 262 historical wheat accessions. 【Result】 The sequences of TaSnRK2.10 on genomes A and D were cloned, named as TaSnRK2.10-A and TaSnRK2.10-D. There was no SNP detected in the sequence of TaSnRK2.10-D, but the full length of TaSnRK2.10-A was 4 688 bp with 15 SNPs and 2 InDels. Among them, 8 SNPs were identified in the promoter region, 2 SNPs in 5′-UTR region, and 5 SNPs in the coding region with 2 SNPs in exon. One of SNPs in exon was non-synonymous mutation. Four markers were developed. They were PM1 and PM2 for the promoter region, GM1 and GM2 for the coding region. TaSnRK2.10-A was mapped in the intervals between markers Xwpt7001 and WMC48 on chromosome 4A, with 5.1 cM and 25.7 cM from the flanking markers. In the natural populations consisted of 262 accessions, four haplotypes of TaSnRK2.10-A were detected by four markers that were associated with 1000-grain weight, spike per plant and spikelet per spike. The HapⅡand HapⅢ of TaSnRK2.10-A are considered as potential superior haplotypes for the improvement of 1000-grain weight. The base C at the site of 4 184 bp is a superior allele for high 1000-grain weight. 【Conclusion】 The present research mapped TaSnRK2.10-A on the chromosome 4A. The HapⅡand HapⅢ of TaSnRK2.10-A are considered as potential superior haplotypes for the improvement of 1000-grain weight, while HapⅣ is a potential superior haplotype for spike per plant. The cytosine (C) at the position of 4 184 bp is the superior allele.

【Objective】 In this work, digital gene expression tag profiling (DGE) was employed to investigate the differentially expressed genes (DEGs) in the embryo of maize seeds treated by a controlled deterioration treatment (CDT) to provide a basis for a better understanding of essential molecular mechanism underlying seed deterioration. 【Method】 In this study, maize (Zea mays L.) cultivar (zhengdan 958) seeds were used as a model and treated by CDT (45℃, 100% relative humidity) for 72 h (T), untreated seeds were used as control (CK). DGE was performed and the high-quality sequences were mapped to the reference genome and maize genes database to obtain the expression genes. The expression level of each gene was calculated by RPKM method. A combination of FDR<0.001 and the absolute value of |log2 ratio (T/CK)|≥1 was used as the threshold to determine the significance of gene expression difference. Finally, GO and pathway enrichment analysis were used to identify the significantly enriched function classification and metabolic pathways in DEGs.【Result】About 3 2000 mRNAs were detected in dry maize embryos (CK). A total of 4 713 DEGs, including 2 874 up-regulated and 1 839 down-regulated, were identified under CDT for 72 h. GO enrichment analysis revealed that the DEGs involved in three GO categories, i.e., cellular component, molecular function and biological process. The proteins coded by these genes were distributed on organelle/membrane in cells and participated in some metabolic processes, signaling transduction, response to stimulus and death process, etc. They would have binding, catalytic activity, and antioxidant activity, etc. There were 2 470 annotated DEGs that participated in 288 KEGG pathways in which 16 pathways were significantly enriched. Among these pathways, there were 113 genes involved in energy metabolism, i.e, 59 genes in glycolysis /gluconeogenesis, 50 genes in pyruvate metabolism, and 31 genes in pentose phosphate metabolism, respectively. The genes encoding enolase and glyceraldehyde 3-phosphate dehydrogenase in glycolysis /gluconeogenesis，pyruvate kinase in pyruvate metabolism，and alpha-L-fucosidase in pentose phosphate metabolism were up-regulated at highest levels. There were 25 genes that regulate the metabolism of NADH (9 up-regulated and 16 down-regulated genes) and 10 genes regulate the metabolism of NADPH (4 up-regulated and 6 down-regulated genes) were detected. They may regulate reactive oxygen species (ROS) production and accumulation. 【Conclusion】DGE provided an innovative and powerful tool for investigating the molecular mechanism of seed deterioration or vigor loss during aging. CDT could affect DEGs expression in dry maize embryos and then energy metabolism in cells. They would inhibit glycolytic pathway and promote ROS production and accumulation, then, accelerate cells aging or death in seed embryos, and ultimately lead to seed deterioration and vigor loss. DEGs might play a critical role in the process.

【Objective】The objective of this study is to fill the gaps in miRNA-related fields of Nicotiana tomentosiformis and N. sylvestris research as quickly as possible, and to reveal the growth and development regulation mechanism in N. tobacum, 【Method】the microRNAs and their target genes of N. tomentosiformis and Nicotiana sylvestris were genome-wide predicted and analyzed. miRNAs were predicted by the method of homologous alignment and secondary structure characteristics of pre-miRNA: reference sequence in sequence alignment of N. tomentosiformis and N. sylvestris were allowed 1-2 mispairings; secondary structure of miRNA was classic stem loop structure, maximum value of MEF was -25, minimum value of MEFI was 0.85, the predicted miRNA and the same family miRNA located on the same arm of hairpin structure; E-values of encoding protein sequence less than or equal to 1e-6 were eliminated. 【Result】 A total 162 miRNAs belonging to 39 families were identified in N. tomentosiformis, including 14 pairs of sense and antisense strand miRNAs and 5 gene clusters. A total 169 miRNAs belonging to 40 families were identified in N. sylvestris, including 13 pairs of sense and antisense strand miRNAs and 3 gene clusters. In high degree of miRNA conservative families, members of the distribution and membership were near in 2 wild tobaccos. While in a relatively low degree of conservative families, members of 2 wild tobaccos differed obviously. Nine families like miR5021, miR5203 and so on, got members in N. tomentosiformis. Ten families like miR1446, miR1509 and so on, got members in N. sylvestris. Antisense miRNA and their sense partners from 2 wild tobaccos differed from 1 to 4 bases, these differences location presented preferences in different families, and the preferences were similar in 2 wild tobaccos: 9th, 12th, 13th base in miR164 family, 1st, 21st base in miR172 family, 2nd, 17th base in miR396 family, 15th, 20th base in miR399 family. Gene cluster of 2 wild tobaccos consisted of miR156 family and miR169 family, distance of pre-clusters was less than 350nt. miR6019/miR6020 gene clusters were found in N. tomentosiformis for the first time. Unigene of N. tabacum was used as target genes. In N. tomentosiformis, 749 target genes of 122 miRNAs were identified. With duplicate genes eliminated, 206 non-redundant target genes were identified, in which, 89 target genes (43%) got GO annotations. In N. sylvestris, 117 target genes of 650 miRNAs were identified. With duplicate genes eliminated, 169 non-redundant target genes were identified, in which, 78 target genes (46%) got GO annotations. In terms of molecular function, most of the target genes have binding activity. In the process of biology, target genes mainly involved in the development process, reproductive process, multicellular organ development process, stress response and so on. 【Conclusion】 In N. sylvestris, there are more target genes control development and multicellular development, while in N. tomentosiformis, there are more target genes control stimulus and press reply.

【Objective】In order to seek the impacts of climate warming on growth process and yield of spring maize in recent 20 years in Northeast China, a study was conducted for providing a theoretical basis for food security.【Method】Three provinces (Heilongjiang, Jilin and Liaoning province) in Northeast China were selected for conducting regional research. Making use of the meteorological observation data and the long-term observation data of spring maize in recent 20 years in Northeast China, the relationships between climate factors (temperature and precipitation) and growth process and historical production data during growing season were analyzed by correlation and regression analysis and other statistical methods.【Result】It was found that the diurnal maximum temperature, minimum temperature and average temperature in Northeast China during spring maize growth season in 1989-2009 were rising significantly. Climate trend rates were 0.050, 0.045 and 0.044 ℃?a-1. During spring maize growth warming amplitude during the daytime was larger than the night. No significant change in the precipitation was found. During 20 years, the change tendency of spring maize average sowing date was 0.10 d in advance every year in experiment stations of Heilongjiang province, but the change tendencies were postponed in experiment stations of Jilin and Liaoning provinces, respectively, 0.18 and 0.21 d per year. The change tendencies of spring maize average maturity date were postponed in experiment stations of Heilongjiang, Liaoning and Jilin provinces, respectively, 0.39, 0.35 and 0.55 d per year. The change tendencies of spring maize average total growth days in Heilongjiang, Jilin and Liaoning provinces were an annual increase of 0.49, 0.17 and 0.34 d, respectively. The amplitude of the delayed maturity date was larger than the delayed sowing date, eventually resulting in increased number of spring maize growth days. The national varieties data and meteorological data correlation analysis in 1991-2006 showed that the rise of the diurnal maximum temperature (Tmax) in Heilongjiang extended national varieties growth period. The correlation between the state-approved varieties growth period and the diurnal maximum temperature (Tmax), minimum temperature (Tmin), average temperature (Tavg) in Jilin and Liaoning provinces showed a positive correlation. Analysis of the relationship between the growth period of the state-approved varieties and experiment station varieties by using T test showed that the growth days of them had a consistent increasing trend in Heilongjiang and Liaoning provinces, and the increasing trend had no significant difference. By adopting linear partial regression test method the importance of varieties and climatic factors on spring maize growth period was analyzed and the results showed that the extension of varieties growth period was the main factor causing experiment station varieties growth period prolonged. The average yield of spring maize showed an increasing trend, and the ascending order of the trend was Liaoning, Jilin and Heilongjiang provinces in recent 20 years. The farther north the yield increasing trend was greater, and the yield increase was more obvious. Panel data analysis showed that changes of the Tavg, Tmax and Tmin affected the yield of spring maize in prefecture-level city of Northeast China in 1989-2009. When the temperature rose, the yield in northeast significantly increased, especially in the Sanjiang plain area of the eastern Heilongjiang province, but the yield in southwest cities was obvious reduced in Northeast China.【Conclusion】Above evidences indicate that spring maize production is affected by climate warming in Northeast China. Spring maize varieties are gradually adapted to climate warming. Varieties improvement and sowing date adjustment for adapting climate warming can improve the yield of spring maize.

【Objective】 Spring corn was the staple crop in Liaoxi area, but the shortage of precipitation and the big precipitation variation and its uneven distribution of time and space are the key limiting factor in the area. The limited rainfall can not meet the demand of stable and high yield of spring corn. Ridge film mulching and furrow seeding can collect natural precipitation effectively and improve rainfall resource availability. The purpose of this study is to make full use of the limited natural rainfall, improve water use efficiency of dry farming area and promote high and stable yield of spring corn.【Method】 Micro- rainwater-collecting planting mode by ridge film mulching and furrow seeding for spring corn was carried out for the sake of studying the effect of different planting patterns on soil water, yield and water use efficiency in 2007-2013. Four treatments were made in this experiment: ridge film mulching and furrow seeding (furrow was uncovered, T1), ridge film mulching and furrow straw mulching (T2), ridge and furrow film mulching (T3) and the traditional planting (CK). The experiment was designed by random grouping and repeated three times. The traditional planting was sowing in furrows, line spacing was 50 cm. Ridges and furrows of ridge film mulching and furrow seeding were in opposite arrangement, furrow width was 80 cm and ridge width was 40 cm, ridge height was 15 cm. The ridge was the rain collecting area. The furrow was the planting area and two lines of corn were planted. Planting density of different treatments was 52 500 plants/hm2. Seed manure was diammonium phosphate（375 kg•hm-2, N 18%, P2O5 46%）and topdressing was urea (375 kg•hm-2, N 46%).【Result】 The results showed that the ridge film mulching and furrow seeding could collect natural rainwater effectively, raise emergence rate of spring corn by 13.0% and 14.9% and emergence time was 1-2 d early in 2009-2010. Under the condition of no effective rainfall for a long time in early days, the order of increase of soil water storage in different treatments was T2＞T1＞CK＞T3 after an effective rainfall. T1 and T2 enhanced water infiltration depth to 60㎝ in planting area compared with the control, which was 40 cm. The runoff efficiency was 61% and the rate of increase of soil moisture storage of T1 and T2 was 72% and 88%. The average of soil water of T1, T2, T3 and the control was 135.27 mm, 140.93 mm, 127.85 mm and 118.98 mm during growth stages of spring corn, increased by 13.69%, 18.45% and 7.46%, respectively, compared with the control. The yield of T1-T3 increased by 24.31%-32.58%, 9.95%-17.81%, 32.12%-37.16%, 16.58%-27.96%, 2.50%-9.40%, 10.85%-29.33% and 4.14%-17.95% in 2007-2013, the average production of T1, T2 and T3 increased by 14.52%, 20.01% and 23.44% compared with the control. The water use efficiency of T1-T3 increased by 24.66%-36.07%, 14.12%-23.73%, 38.34%-53.89%, 29.07%-35.68%, 1.20%-19.60% and 9.02%-32.55%, the average of water use efficiency of T1, T2 and T3 increased by 20.39%, 27.94% and 28.02% compared with the control. It was also found that the effect of increase of yield and water use efficiency was significant under lower rainfall.【Conclusion】 The effect of Micro- rainwater-collecting planting mode by ridge film mulching and furrow seeding in dry farmland on rainwater harvesting, water storage and soil moisture conservation was definite after 7 years of study in semi-arid region in Liaoxi area. The technique could mitigate the negative impact of drought on spring sowing and seedlings growth in spring and enrich the theoretical basis of the rain-harvesting agriculture of dry farming in the area. By application and dissemination of the technique, it could improve rainfall resources utilization and water use efficiency and promote stable and high yield of spring corn and healthy and sustainable development of dry farming in the area. The study will provide important references for development of the rain-harvesting agriculture of dry farming in the north of China.

【Objective】 The response of N2 fixation and yield to N fertilizer, even with same N application rate, gives varying and contradicting results. The contradicting results depend on the perception of the system of soybean nitrogen fixation and root to the soil inorganic N concentration. The nodulation, fixation and yield of soybean with different soil inorganic N concentrations were studied to find the reasonable soil inorganic N concentration for promoting soybean yield, nodulation and fixation. i.e. to master the quantified relationship between soil inorganic N concentration, nodulation and fixation. Then it will have a significant theory and production meaning for adjusting N application rate, timing, and forecasting the effect of N fertilizer on the fixation and yield. 【Method】A pot experiment was conducted to study the effects of different soil inorganic nitrogen concentrations, which derived from total fertilizer N (as urea) applied at vegetative stage (V2), at early blooming stage (R1), at early podding stage (R3) and at grain filling stage (R5), on the ability of N2 fixation and yield, by measuring the nodule number, biomass and nitrogenase activity and yield components. Through the research, the response of soybean nodulation and yield to soil inorganic N concentration cleared, and thus mastering the quantity relationship between soil inorganic N concentration and N application, nodulation and yield, and then providing a theoretical basis for agriculture production and scientific research. 【Result】 The nodule number, biomass and nitrogenase activity of soybean reached peak at R4 stage, while decreased at R6 stage, and the nodule number, biomass and nitrogenase activity were in order of V2＞R5＞R3＞R1. The nodule biomass decreased by 15%, 18%, 17% and 32% compared to that of CK. The nodule number decreased by 13%, 18%, 19% and 20%. The nitrogenase activity decreased by 19%, 22%, 23% and 32%. Meanwhile, there were significant negative linear correlations between nodule number, biomass and nitrogenase activity at R6 stage and the concentration of soil inorganic nitrogen. Moreover, The trends of dry matter accumulation and yield were expressed as an order of R1＞R3＞V2＞R5. Except the treatments at R5 had no significant correlations with the treatment of CK, the other treatments had a significant promoted effect on dry matter accumulation and yield. The treatment of N3 had the highest dry matter accumulation, plant height, pod number, pod dry weight and yield than the other treatments at different growth stages. The soil inorganic N concentration at V2 stage had a significant effect on nodulation and the ability of nitrogen fixation, and that at R1 stage had a significant effect on growth and yield. There were different threshold values of soil inorganic nitrogen concentration at different growth stages. The soil inorganic N concentration should be reached 135.8 mg•kg -1 at V2 stage, 58-91 mg•kg -1 at R1 stage; 29.4-62.8 mg•kg -1 at R3 stage; 102.3 mg•kg -1 at R5 stage to promote soybean nitrogen fixation and yield. 【Conclusion】 The response of N2 fixation and yield to N fertilizer mainly depended on the soil inorganic N concentration which was affected by N fertilizer application rate and timing. The soil inorganic N concentration was adjusted by agriculture production and scientific experiment. The nodule number, biomass and nitrogenase activity were significantly affected by the soil inorganic N concentration at V2 stage than that at the other growth stages, while the biomass and yield were significantly affected by the soil inorganic N concentration at R1 stage than that at other growth stages.

【Objective】 Endo polygalacturonases (endoPGs) is considered to be one of the important pathogenic factors. The objectives of this study are to clone and compare endoPGs from the strong pathogenic strain YC-9 and the weak pathogenic strain LF-2 from Rhizoctonia solani, analyze the sequence features, evolutionary relationships and expression characteristics of endoPG1 and endoPG2, and to provide a theoretical foundation for clarifying molecular mechanism of the pathogenicity. 【Method】 Degenerate primers were developed based on the sequence of different endoPGs from plant pathogens in GenBank, and partial cDNA fragments of the strains YC-9 and LF-2 were firstly acquired, then the full-length cDNA sequences were cloned through RACE techniques, and the conserved domains and sequence features of the genes were analyzed by bioinformatics’ methods. A phylogenetic tree was constructed using the MEGA 4.0 software, and the relative expression characters of endoPG1 and endoPG2 were also analyzed by real-time RT-PCR. 【Result】 Two endo polygalacturonases genes were cloned and named as endoPG1 and endoPG2. The genes possessed the conserved domains of PLNO3003 gene superfamily. The open reading frame (ORF) of the full length cDNA was 1 086 bp, encoding a protein of 361 amino acid residues. Differences between endoPG1 and endoPG2 were indicated by comparing the full-length cDNA sequences and transmembrane regions of prediction protein. The PLNO3003 subunit phylogenetic tree was constructed, which showed that the corresponding amino acid sequences of endoPGs from R. solani formed an independent branch, and that of endoPG1 and endoPG2 had the highest homology. Real-time RT-PCR demonstrated that the expression of endoPG1 and endoPG2 were more obviously up-regulated in hyphae of inoculated tobacco leaves than in hyphae of non-inoculated ones, and the expression quantity of endoPG1 was higher than that of endoPG2. 【Conclusion】 The full cDNA sequences of endoPG1 and endoPG2 were successfully cloned from tobacco target spot disease of R. solani, and both have conserved domains of PLNO3003 gene superfamily, and it has obviously differences between transmembrane regions of prediction protein of endoPG1 and endoPG2. Phylogenetic analysis showed that the corresponding amino acid sequences of endoPG1 and endoPG2 have a highest homology relationship. Finally the endoPG1 and endoPG2 gene expression can be obviously induced by interaction with tobacco and the strong and weak pathogenicity strains in YC-9 and LF-2 have obvious differences.

【Objective】Based on the cloning of a carboxylesterase gene of Bactrocera dorsalis, the expressions of this gene among different developmental stages, tissues, and under insecticide stimulation were analyzed. Combined with previous data of enzyme activity assay and synergist analysis, the objectives of this study are to clarify the reaction of carboxylesterase when stimulated by β-cypermethrin, and lay a foundation for the future study of the mechanism that how carboxylesterase works in the detoxification of pyrethroid insecticides.【Method】From the transcriptome data of B. dorsalis, a partial sequence of carboxylesterase gene was screened out by using homologous blast, and the full sequence was cloned by using RACE technology. The ORF, deduced amino acid sequence, and molecular weight were predicted, and a phylogenetic tree with carboxylesterase genes from other insects was constructed by using maximum likelihood method to clarify its molecular characterization. RNA was extracted from different developmental stages and the midgut, Malpighian tubules, fat body, trachea of the 3rd instar larvae. Based on the reference evaluation, qPCR was used for the expression analysis of different developmental stages, tissues, and stimulated by insecticide. 【Result】A partial sequence of 1 028 bp was found in the transcriptome data. The results of 5′ and 3′ amplification were 1 073 and 625 bp, respectively. After sequence assemble and verification, the full-length of this gene was 1 872 bp, contained a complete ORF of 1 710 bp, and encoding a protein of 569 amino acids with a predicted molecular mass of 63.3 kD. It was named as BdCAREB1 and submitted to GenBank with an accession number of KF539980. The phylogenetic tree of BdCAREB1 showed high homology with carboxylesterase of Diptera. The qPCR result of different developmental stages of the 3rd instar larvae showed BdCAREB1 was highly expressed in the 3rd instar larvae, followed by the adults, and it was the lowest in the eggs. Among different tissues, the BdCAREB1 was highly expressed in fat body. The expressions between midgut and Malpighian tubules were quite the same, and in trachea, the expression was the lowest. After the insects were fed on a diet containing 0.33 μg?g-1 (insecticide/diet) β-cypermethrin, the expression of BdCAREB1 was significantly up-regulated in larvae and fat body. 【Conclusion】The specific expressions of BdCAREB1 were identified in different developmental stages and tissues, which were highly enriched in the larvae and fat body. It showed inducibility by the stimulation of β-cypermethrin. Combined with previous data of enzyme activity assay and synergist analysis, BdCAREB1 may be involved in the detoxification of β-cypermethrin.

【Objective】The objective of this study is to elucidate the molecular function of the three cytochrome P450 genes of CYP4 family from the locust (Locusta migratoria). 【Method】According to the predicted P450 sequences from ESTs database of locust, the primers were designed, the full-length sequences of three novel genes were amplified and named CYP4C69, CYP4C73 and CYP4DH1 (GenBank accession numbers: KF857162, KF857163, KF857164), respectively. The mRNA expression levels of CYP4C69, CYP4C73 and CYP4DH1 were analyzed in different developmental stages and tissues of locusts by real-time quantitative PCR. The RNAi ef?ciency of dsCYP4C69, dsCYP4C73 and dsCYP4DH1 was evaluated by real-time quantitative PCR. The toxic effects of malathlon, chlorpyrlfos and deltamethrin on 2nd-instar nymphs were analyzed and the LC30 doses of the three insecticides were calculated by SPSS software. As CYP4C69, CYP4C73 and CYP4DH1 transcripts were signi?cantly repressed in locusts by RNAi, the susceptibility of the dsRNA injected locusts to different insecticides, such as malathion, chlorpyrifos and deltamethrin were assessed.【Result】The full-length cDNA sequences of CYP450 genes including CYP4C69, CYP4C73 and CYP4DH1 were obtained by RT-PCR. The cDNA sequences of CYP4C69, CYP4C73 and CYP4DH1 had open reading frames of 1 518, 1 521 and 1 512 nucleotides which encoded proteins of 506, 507 and 504 amino acid residues, respectively. Moreover, the expression level of CYP4C69 was higher in 5th-instar nymph than in the adult stage. However, CYP4C73 and CYP4DH1 expressed during the whole developmental stages and were relatively higher in 1st-, 4th- and 5th-instar nymphs and adult stage. The results of CYP4 genes expression from different tissues indicated that all of the three genes had a high expression level in gastric caeca, CYP4C69 and CYP4DH1 highly expressed in fat body. It was found that CYP4C69, CYP4C73 and CYP4DH1 transcripts were signi?cantly repressed in the locusts after dsRNA injection. The toxic effects of malathion, chlorpyrifos and deltamethrin on 2nd-instar nymphs indicated that the LC30 doses of the three insecticides were 59.438, 8.215, 0.759 μg•mL-1, respectively. The susceptibility of locusts to malathlon, chlorpyrlfos and deltamethrin showed that there was no marked difference between locusts’ injection of dsCYP4C69, dsCYP4C73, dsCYP4DH1 and dsGFP. 【Conclusion】The full-length cDNA sequences of CYP4C69, CYP4C73 and CYP4DH1 were obtained. And these genes could be silenced by dsCYP4C69, dsCYP4C73 and dsCYP4DH1. However, treatment with dsRNA of CYP4C73, CYP4C69, CYP4DH1 and dsGFP did not show signi?cant effects on the susceptibility of the locusts to malathion, chlorpyrifos and deltamethrin.

【Objective】On the issues of drip irrigation water consumption and rational application of water and fertilization in the winter wheat belt region of Huang-Huai-Hai Plain, no-tillage fields were chosen as experimental sites located in Huantai, Shandong Province to carry out a systematic study on the effects of drip fertigation on soil moisture vertical migration, winter wheat grain yield and its components and water use efficiency (WUE).【Method】The study adopted the methods of irrigation recharge by measuring soil moisture and drip fertigation in the main growth period compared with conventional flood irrigation and fertilization treatment. Five drip irrigation level treatments including 65 mm (W1), 98mm (W2), 130 mm (W3), 195 mm (W4) and 260 mm (W5) were designed. The corresponding NPK ratio was set up at tillering stage, joining stage, booting stage, young flowering and filling stages of winter wheat under the 130mm of irrigation treatment level and 3 factors of NPK and 4 levels quadratic saturation D-optimization design were adopted for field experiment. The 4 levels of N, P, K were that “0 level” with 0, 0, 0, “1 level” with 94.5, 42.4 and 59.2 kg•hm-2, “2 level” with 189, 84.7 and 118.3 kg•hm-2, “3 level” with 270, 121 and 169 kg•hm-2.【Result】The results of experiment showed that the more drip irrigation amount is, the deeper soil moisture vertical migration will be, the soil moisture vertical migration of W1, W3 and W5 treatments moved to 60, 80 and 100 cm, and could get risk of water percolation while drip irrigation level achieved 260 mm. Soil moisture content of W1 treatment was obviously lower than others, 130 mm and above drip irrigation treatment made soil moisture content at 0-80 cm soil layers over 75%-80% of the field moisture capacity during the growth period. Compared with conventional flood irrigation and fertilization treatment, the drip fertigation significantly increased the effective panicles of winter wheat. The drip irrigation amount had a positive correlation with grain number per spike and a negative correlation with 1000-grain weight in different drip irrigation treatments. The grain yield was the highest when irrigation amount was 130 mm. Drip irrigation apparently increased water use efficiency (WUE) and with the highest of W3 treatment that was 2.28 kg•m-3. Experimental fitting results indicated that the optimal fertilizer amount of N, P2O5 and K2O in pilot area was 206.63, 86.72 and 88.07 kg•hm-2. 【Conclusion】It was concluded that the winter wheat in no-tillage fields by irrigation recharge by measuring soil moisture and drip fertigation significantly increased, WUE and yield by 57.46% and 21.13% in Huang-Huai-Hai Plain, respectively, compared with conventional treatment. The main reason was that the water moved to crop roots area after drip irrigation that could reduce the risk of water percolation and promote fertilizer-absorbing with water. The rational ratio of drip fertigation could totally make water saving by 51.85%, N, P, K fertilizer saving, respectively, by 23.47%, 28.33% and 47.89%.

【Objective】 This study aims at comprehensively evaluate different winter green manures on rice growth dynamics and paddy field weed community in southern rice region for further optimization of rice cropping patterns, and development of winter crop planting and development. 【Method】 From winter in 2011 in double cropping area in southern Jiangxi Agricultural University Science Park, winter green manure species including legumes, grasses, and cruciferous were selected to make comparison of milk vetch, rapeseed, ryegrass and mixed of green manure (vetch × rapeseed × fertile turnip) on rice yield and its components, dry matter production characteristics and weed communities. 【Result】 The results showed that the dry matter weight of population of rice in the treatment of MV-R-R(milk vetch-rice-rice) are all higher than others at tillering stage, booting stage, heading stage and maturity. The rate of increase was 14.46%, 10.99%, 11.83% and 7.23%, respectively. Compared with RG-R-R (ryegrass-rice-rice), RP-R-R (rapeseed-rice-rice), MS-R-R (mixed green manure-rice-rice), rice yield of MV-R-R was higher, increased by 6.61%, 3.29%, and 0.78%, respectively. Among the 4 different winter green manures, rice yield and yield components (effective panicles, grain number per panicle, seed-setting rate and 1000-grain weight) in treatment of MV-R-R were higher than other treatments. Dry matter accumulation in treatment of MV-R-R was higher than others in main growth periods of rice except for heading to maturity. Compared with RG-R-R, RP-R-R, MS-R-R (mixed green manure-rice-rice), the increasing rate were 11.38%-17.72%, 4.50%-28.00%, and 1.85%-35.25%, respectively. The dry matter accumulation in the periods of sowing to tillering and booting to heading were the biggest, which is the stage of rapid accumulation of substances. In the four treatments, dry matter accumulation at the two growth stages reached 70.43%, 60.50%, 69.87%, and 69.08% of that at maturity. While the proportion of dry matter accumulation from heading to maturity was the lowest. Analysis of dry weight ratio of stem-sheath, leaf, and panicle to total plant at middle and late stages of rice, the dry matter ratio of stem-sheath was the biggest at booting stage. But they continued to decrease with the growth, reached a minimum at maturity, and in each treatment decreased from 58.30%-64.20% to 27.95%-30.47%. Dry matter ratio of leaf was the biggest at booting stage, reduced from 35.80%-41.70% to 13.07%-14.62%. According to the survey, weeds in paddy field belong to seven families, eleven species. The weed densities of Echinochloa crusgalli, Monochoia vaginalis, Sagittaria pygmaea and Eleocharis yokoscensis were greater than others, accounting for 80.84% of field weeds. The treatment of MV-R-R had a significant effect on paddy weed density and the effect on inhibiting weeds was obvious better than other treatments. The treatment of RG-R-R was the lowest. By analyzing the relative abundance of weed species, the following conclusions could be drawn. The basic weed community compositions of the MV-R-R and RG-R-R were Monochoia vaginalis+ Echinochloa crusgalli+Eleocharis yokoscensis, but that of the RP-R-R and MS-R-R were Monochoia vaginalis+ Echinochloa crusgalli+ Sagittaria pygmaea. The species richness of the RG-R-R were maximum, significantly higher than other treatments. For the Shannon index, the species diversity of the treatment of winter planting-ryegrass was the highest, while the MV-R-R was the lowest. From the point of view of Simpson index, the weed dominance of MV-R-R was the highest, while the RG-R-R was the lowest. From the point of view Pielou evenness index, the weed evenness of the MV-R-R was higher than other treatments, but the difference was not significant. While the weed evenness of the other winter planting-green manure pattern decreased, of which the RG-R-R was the lowest.【Conclusion】In conclusion, the treatment of milk vetch-rice-rice can ensure higher dry matter accumulation and yield potential, which have a significant impact on weed species and density. Also it can help to improve weed uniformity and decrease the harm of dominant species of weeds in paddy field.

【Objective】 The objective of this study is to identify the cucumber auxin response factor (ARF), forecast the small RNAs and verify the relationship between ARF with small RNAs and auxin. The expression pattern of ARF during seed germination was analyzed and the effect of ARF on the process of parthenocarpy and seed germination was deduced. 【Method】 The Arabidopsis and rice ARF proteins were used to search the cucumber genome database. Then, the retrieved cucumber ARF family structure was analyzed and the small RNAs were predicted. The predicted gma-MIR160o precursur was built into the pCAMBIA2301. By Agrobacterium-mediated method, it was imported into parthenocarpy cucumber varieties. Transgenic plants were validated by RT-PCR. Using real-time RT-PCR method, the expression patterns of ARF were analyzed in auxin treatment, flowing and seed germination. 【Result】 By comparison with the Arabidopsis and rice ARF protein sequences, there are 18 cucumber ARF protein sequences. ARF proteins were divided into four classes. The number of exons varied from 2 to 18, and the structure was similar among the same class. Phylogenetic tree showed that the similarity of 18 genes was not high. Small RNAs corresponds to ARF genes in cucumber were found. Csa010564, Csa011935, Csa015176, Csa020560 and Csa022361 were miR160 targets. Transgenic test further exhibited mRNA abundance of Csa010564, Csa011935 and Csa015176 decreased, while Csa020560 and Csa022361 rose slightly, indicating that Csa010564, Csa011935 and Csa015176 were miR160 targets. The results of real-time RT-PCR exhibited that mRNA abundance of Csa007296, Csa011935 and Csa015176 in roots, stems and leaves were higher than wild type, indicating the expressions of these genes were positively regulated by auxin. Meanwhile, the expressions in leaves and female flowers were decreased on the second flowering day, while the expression of the ovary was the opposite, especially Csa011935 and Csa015176, indicating that ARF genes play a crucial role in the development of the ovary. In the experiment of the auxin treatment, the expressions in different tissues were up-regulated. The expression of ARF genes in the seed germination was analyzed. The results showed that the expression peak of most genes were 12 h and 48 h in imbibition. 【Conclusion】 The ARF genes were regulated by auxin and the corresponding small RNAs. ARF genes may play a key role in the parthenocarpy and seed germination of cucumber .

【Objective】 This study was conducted to evaluate the effects of aeration cultivation on root architecture and nitrogen metabolism of young peach trees in order to lay a foundation for finding out the role of oxygen in the peach tree root growth and development.【Method】Two-year-old peach tree ‘Chunmei’ was selected to study the effects of aeration cultivation on soil O2 content, root activity, root architecture and nitrogen metabolism of young peach trees. Plastic box with uniform small holes at the bottom was used as the aerobic cultivation device. There were two treatments including the control (non-aeration) and aeration cultivation. The change of soil O2 concentration, root activity, root total nitrogen and root transaminase activity was determined in the peach tree growing season. The professional edition of root analysis system WinRHIZO was used to determine the parameter of root architecture after the new shoots slightly stop growing.【Result】Aeration cultivation improved soil O2 content. After aeration cultivation, the total root length, total root surface area, number of first lateral roots and second lateral roots, number of tips, total root volume, number of forks, number of cross increased by 50.53%, 42.66%, 13.64%, 18.51%, 44.94%, 5.48%, 37.01% and 54.41%, respectively compared with the control. The average length of first lateral roots and root average diameter decreased. The angle between first lateral roots and vertically downward direction was decreased, the number of small angle increased, and root tended to vertically distribution under aeration cultivation. The dry matter of roots which the diameter was less than 2 mm was significantly higher than control and the dry matter of roots which the diameter was greater than 5 mm was decreased under aeration cultivation. The activity of root system, the activities of NR, GOT, GPT and the content of root total nitrogen were higher than the control under aeration cultivation and they increased 19.04%, 29.80%, 6.56%, 19.91% and 18.90%, respectively. After aeration cultivation, the dry matter accumulation and stem of young peach trees were higher than the control. 【Conclusion】 Aeration cultivation can increase the soil O2 concentration and promote the occurrence and growth of fine roots. It can induce root straight down growth and improve the level of the root nitrogen metabolism. It is advantageous to accumulate the dry matter and promote the growth and development of young peach trees.

【Objective】The relationship between myofibril characteristics and meat quality of breast of different chicken varieties was investigated. The purpose of the present study was to provide a scientific basis for selection of special varieties and understanding of the effect of raw meat quality on the quality of meat products. 【Method】The microstructures and ultrastructures of breast muscle myofibril of eight Chinese native chickens were observed by scanning electron micrograph (SEM) and transmission electron microscope (TEM) , the quantitative analysis of myofibril characteristics was conducted by Image-Pro Plus 6.0, at the same time cooking loss, shear force, texture profile analysis (TPA) and myofibril fragmentation index (MFI) were measured. The correlation between myofibril characteristics and processing features was established by correlation analysis. The Statistic Analysis System 9.2 (SAS9.2) was used to analyze the variance, multiple comparison and correlation.【Result】Eight Chinese native chickens showed a significant difference in basic chemical compositions, colour, pH, tenderness, water holding capacity, texture properties, fibre diameter, fibre density, sarcomere length and MFI (P＜0.05). The water content of Silkie was significantly higher than the other varieties (P＜0.05) , and the Chai hens’ was the lowest. The protein contents of eight Chinese native chickens were ranging from 22.03% to 24.53%, and the extent of variation was minor. The protein content of Princess chicken was the lowest, but its fat content was the highest. The lightness and redness of Bantam were significantly higher than the other varieties (P＜0.05). The pH of Silkie was the highest. The shear forces of Princess chicken, Tsingyuan’s chicken, Silkie and Spring chicken were lower than the other varieties and the cooking loss of Silkie was the lowest. The fibre diameters of Princess chicken, Tsingyuan’s chicken, Silkie and Spring chicken were thinner, the fibre densities were denser; the sarcomere length of Tsingyuan’s chicken was the longest. MFI of Sanhuang chicken and Princess chicken were higher than the other varieties. The correlation analysis showed that the fibre diameter was positively correlated (P＜0.05) with shear force, but the fibre density was negatively correlated (P＜0.05) with shear force, the sarcomere length was negatively correlated (P＜0.05) with cooking loss.【Conclusion】There was a significant correlation between myofibril characteristics and meat quality, the myofibril characteristics had an important effect on meat quality.

【Objective】This study was designed to investigate the influence of heating on myofibrillar proteins(MP) secondary structure and gel properties, and to reveal the relationship between MP secondary structure and gel properties.【Method】Forty-day-old commercial AA broilers were slaughtered. The breast muscle was stored at -18℃ before MP was extracted. The MP secondary structure was measured using a circular dichroism spectra to determine the content of α-helix, β-sheet, β-turn and random coil during heating. The values of G' and Tanδ were continuously measured using a rheometer during heating. The influence of heating temperature on textural properties of MP gel prepared under different temperatures was measured using a textural analyzer. Spin–spin relaxation time (T2) of the gels prepared under different temperatures was measured using a NMR Analyzer in order to investigate the water distribution of gels. SPSS17.0 software was used to analyze the data such as correlation analysis so as to illustrate the relationship between the heating temperature and protein structure and gel properties. 【Result】 Heating temperature influenced significantly MP secondary structure. The α-helix content declined from 95.77% to 45.05% as temperature increased from 30℃ to 80℃. The α-helix content declined slightly as temperature increased from 30℃ to 40℃ and from 70℃ to 80℃, declined abruptly between 40℃ and 70℃ (P＜0.05). The β-sheet content increased from 0.20% to 12.65% as temperature increased from 30℃ to 80℃. The decline in α-helix content indicates the unfolding of a protein molecule. The increase in β-sheet content indicates the aggregation of unfolding protein molecules. Heating temperature influenced rheological properties, textural properties and water distribution of MP. G' values began to increase at about 42℃ indicating the starting of protein gelling. G’ values showed a sharp increase between 42℃ and 50℃ (177 Pa) with a subsequent decrease between 50℃ and 55℃ and a final increase between 55℃ and 75℃. Hardness values of MP gel increased as the temperature of the gel prepared rose from 40℃ to 75℃ and reached the maximum value of 51.4 g at 75℃. Springiness values reached the maximum value of 0.754 at 55℃. T2 curves of MP gel usually contained 3 peaks and T22 means immobile water. T22 values of the gels decreased from 403.7 ms to 265.6 ms as the gel preparing temperature rose from 40℃ to 60℃, which indicated that water mobility decreased as temperature rose from 40℃ to 60℃. Heating temperature and β-sheet content showed a significant positive relationship to G' and hardness of gel (P＜0.01), and the correlation coefficients were all over 0.849. The correlation analysis indicated that heating caused MP molecules unfolding, aggregating, gelling, and led to significant change of G' and hardness of MP gel. α-Helix and β-sheet, which didn't show a significant relationship to springiness and T22 of the MP gel. It was found that heating led to MP molecules unfolding, aggregating, gelling simultaneously at temperature over 40℃ by analyzing the influence of heating temperature on α-helix, β-sheet content and G’. The unfolding MP molecules rearranging to β-sheet was the key factor for the increase of G’ values. The unfolding MP molecules rearranging to β-sheet was also the key factor for the increase of hardness values by analyzing the influence of heating temperature on β-sheet content and hardness values of MP gel.【Conclusion】The α-helix content declined and β-sheet content increased as temperature increased from 30℃ to 80℃. Heating led to significant changes of protein secondary structure. G' values began to increase and the values of Tanδ began to decrease at about 42℃, which indicated the starting of protein gelling. Gel hardness values reached the maximum 51.4 g at 75℃. Heating temperature and β-sheet content showed a significant positive relationship to G' and hardness of the gel. The increase of β-sheet content was the key factor for the increase of G’ and hardness values of the MP gel.

【Objective】 The morphological, anatomical, cytogenetical and histopathological study in a true hermaphrodite pig was conducted, which aimed at elucidating the basis of genetics and reproductive development, optimizing germplasm resources for pig production. 【Method】 A large white hermaphrodite pig at five months age was used as the experimental group, versus three littermate normal male and female pigs were used as control group. The morphology of external and internal reproductive organs was examined. SRY gene was tested by PCR. Karyotype analysis was performed on metaphases obtained from conventional peripheral blood lymphocyte cultures, and was G banded and C banded. Venous sex hormones including testosterone (T), estradiol (E2), luteinizing hormone (LH), follicle-stimulating hormone (FSH), progesterone (PRG) and prolactin (PRL) were tested using radioimmunoassay. Gonads (2 cm×2 cm) were fixed with 10% neutral formaldehyde, routinely prepared as conventional paraffine sections, and then stained with haematoxylin and eosin (HE stain). The histological sections were observed and documented in light microscope. Hypothalamus and pituitary (2 mm×2 mm) were fixed with 2.5% glutaraldehyde, and ultra structure analysis was observed under the electron microscopic sections. 【Result】 The external genitals had female phenotype with an enlarged clitoris, the anus was joined with the vulva and a testis with well-developed scrotum was on the left in the hermaphrodite pig. In the hermaphrodite pig and female pigs, the diploid chromosome number was 38, XX, karyotype was 10sm+4st+12m+12t, and SRY gene was negative. No differences were found in G-banding pattern and most C-bandings with round pattern were occurred on chromosomes 1, 3, 4, 6, 8, 9, 10, 13, 14 and 16. The chromosome relative length and the arm ratio showed no significant difference (P＞0.05) between the experimental group and the female pigs. The values of T and E2 in hermaphrodite pig were at middle level compared with that in the normal male and the female pigs, but LH, FSH, PRG and PRL were relatively higher, and the ratio of LH/FSH was obviously lower than that in the female pigs. After dissection the hermaphrodite pig, the gonads were shown both ovotestes, the left one with an epididymis, showing the regressive uterine body, uterine horns and vagina. A phallus was hidden in clitoris. Ovotestis was mixed with testis and ovary, showing regressive columnar epithelial cells, normal leydig cells, no spermatogonium and its debris. Epididymis was observed with normal pseudostratified ciliated columnar epithelium cells and no spermatozoal content or cell debris in the lumen. Endometrium was shown degeneration of endometrial lining combination with some of the normal endometrial glands. Secretory cells of hypothalamus and pituitary gland were more developed in the experiment group than that in the control group. Gonadotropin-releasing hormone (GnRH) neures were shown large nucleus, increased and fused organelle. Pituitary was observed mainly gonadotroph, of which quantity was increased and secretory granules were with different sizes in the hermaphrodite pig than that in the normal pigs. The quantity of lactotrophs and the electron density of secretory granules was larger in hermaphrodite pig than that in the female pigs. 【Conclusion】 These results indicated that the intersex pig belongs to a true hermaphrodite pig with no chromosome translocation, sex hormone disturbance and no breeding value.

【Objective】 The objectives of this study are to examine microbial communities from the digestive tract of Bombus lantschouensis reared in the laboratory, and to analyze the specific bacteria spatio-temporally in different developmental stages of B. lantschouensis, which is the important foundation and basis for further studying the function of symbiotic bacteria in bumblebee. 【Method】The fragment of the bacterial 16S rDNA gene was PCR-amplified with the universal eubacterial primers 774F and 1391R and a 16S rDNA clone library of gut bacteria was constructed. Each single clone was picked and sequenced. The sequences were checked for chimeras, sequences of chimeric origin were removed from further analysis. Sequences obtained were analyzed by BLASTn and matched with valid reference sequences in the NCBI (National Center for Biotechnology Information) to determine the bacterial species types. The 16S rDNA gene primers of the specific bacteria Gilliamella apicola and Snodgrassella alvi were designed based on sequencing of bacterial taxonomic groups obtained from the clone library, then constructed recombinant plasmid using the specific bacteria primers and diluted the plasmid to 10 folds series for 5 concentration gradients after concentration determination. Total genomic DNA was extracted from egg, larva, pupa, and 0, 5th, 10th, 15th, 20th days worker bees. The bumblebee β-actin gene was selected as the reference gene, the quantitative real-time PCR (qPCR) was used to explore the quantity variation of symbionts during the different developmental stages of B. lantschouensis.【Result】A total of 213 clones were picked and sequenced, after Chimeras checking, 202 sequences were obtained and then divided into 16 OTUs. The sequence similarity with GenBank sequences was between 93%-99%. The phylogenetic analysis found that the sequences fell into 6 major lineages within the domain bacteria. Among them, 45% and 30% were respectively G. apicola and S. alvi, 10% was Bifidobacterium, 5% was Fructobacillus fructose, 2% was Lactobacillus, 2% was Flavobacterium aciduliphilum, and 6% was mixture of uncommon bacteria types. The qPCR result showed that G. apicola and S. alvi were detected in all development stages, the two bacterial species were similar in the quantity variation patterns with decreasing after increasing, finally reached the steady state. The quantity of G. apicola and S. alvi in egg, larva and pupa was very low, in 5th day significantly higher than those in other development stages, and then gradually reduced, stabilized after 15th day, the bacteria quantity in 20th day showed no difference with 15th day.【Conclusion】Four common symbiotic bacteria including G. apicola, S. alvi, F. fructose and Bifidobacterium were identified in the digestive tract of B. lantschouensis reared in laboratory. G. apicola and S. alvi are dominant bacterium among them. G. apicola and S. alvi both have horizontal and vertical transmission. The quantity of G. apicola and S. alvi in egg, larva and pupa was very low, and increased rapidly after worker bees emergence, stabilized after 15 days. The temporal variation of two dominant bacteria in B. lantschouensis may be related to the function of symbiotic bacteria in bumblebee.

【Objective】Salt tolerance of 278 naked oat accessions from different ecological zones at germination stage were comprehensively evaluated in order to provide a guide for identification and evaluation of salt tolerance of naked oat, and supply abundant materials of salt tolerance for breeding of salt tolerance as breeding parents.【Method】Identification for salt tolerant germplasm of naked oat was conducted in an artificial climate incubator with germination method on paper in petri dish under 1.2% NaCl aqueous solution and distilled water, respectively. The culture condition was constant temperature 25℃, relative humidity (70±5)%, 12 h light (6:00-18:00), light intensity 150 μmol?m-2?s-1 in the incubator. When the length of radicle was equal to that of kernel at least, and the shoot height was not less than 1/2 of seed length, the seed was considered as germinated. Germination potential of tested accessions was counted after 96 hours cultivation. Final germination rate, length of the longest primary root and seedling height were measured after 168 hours cultivation. These four traits were used to identify salt tolerance of tested accessions. Based on salt tolerance coefficients derived from four identification indices, the weight of each index was allocated according to variation coefficient of membership function value of four salt tolerance coefficients, and 278 naked oat accessions were evaluated for salt tolerance with the weighted membership function method and classified through cluster analysis.【Result】Compared with distilled water control, germination potential and germination rate of all accessions were declined except SHX88 and NM47, and the root length and seedling height of all accessions were suppressed by salt stress. There were differences among tested accessions based on the four kinds of identification indices. With the weighted membership function method and cluster analysis, salt tolerance of 278 naked oat accessions were comprehensively evaluated and categorized into five grades. Seventeen accessions were highly salt tolerant, 114 accessions were salt tolerant, 106 accessions were middle tolerant, 25 accessions were susceptible and 16 accessions were highly susceptible. Germination potential and germination rate were closer with salt tolerance at germination stage among the four identification indices, but germination rate was more variable than germination potential among tested accessions.【Conclusion】Germination rate is an efficient identification index for rapid identifying and preliminary screening of salt tolerant accessions. The weighted membership function method has good application value in the comprehensive evaluation of tolerance of naked oat to salt stress. SHX75 and other sixteen naked oat accessions which are highly tolerant to salt stress have provided a basis for breeding, cultivation and mechanism research of salt tolerance.

【Objective】In order to provide a scientific basis for high water-fertilizer use efficiency of rice, the objective of this experiment was to study the effects of water and nitrogen (N) management patterns on population quality and yield of hybrid rice.【Method】Gangyou 527, an elite indica threeline hybrid rice cultivar widely planted in rice production in China, was used, which has high grain yield, extensive adaptability, and pest-resistance with growth duration of 145-150 days from sowing to maturity. The experiment was conducted to optimize N application, understand how different combinations of water and N management affect population quality of hybrid rice and yield, and study the correlations of population quality index and grain yield. Three irrigation regimes including submerged irrigation (W1), controlled alternate irrigation (W2), and dry cultivation (W3) combined with four N application managements at 180 kg?hm-2 of total N conditions were designed in the study. The four N application ratios were applied on different splits: (a) 2 splits: 70% basal and 30% 7 d after transplanting (DAT) (N1), (b) 3 splits: 50% basal, 30% 7 DAT, and 20% panicle N-fertilizer according to different leaf ages when 4th leaves emerged from the top (N2), (c) 4 splits: 30% basal, 30% 7 DAT, 40% panicle N-fertilizer equally when 4th and 2nd leaves emerged from the top (N3), (d) 4 splits: 20% basal, 20% 7 DAT, 60% panicle N-fertilizer equally when 4th and 2nd leaves emerged from the top (N4), in addition, no N application (N0) was designed as control. 【Result】 The results showed that there was an obvious interaction between irrigation regime and N application on dry matter accumulation and leaf area index (LAI) at the mainly growth stages, grain-leaf ratio at heading stage, net photosynthetic rate and yield. Decomposition analysis of water and nitrogen interaction effects showed that appropriate increase of N application ratio of later stage or W2 treatment had a positive effect on yield, the N application treatments had significant differences, and ranked as N3＞N2＞N1, and N4 treatment or W3 treatment resulted in a negative effect of water-nitrogen interaction. Compared to irrigation regimes, the results of water and nitrogen interaction showed that W2 treatment promoted the efficiency of N fertilizer, and achieved the purpose of promoted fertilizer by water. W2 treatment with suitable treatment of N3 application (30% basal, 30% tillering, 40% panicle N-fertilizer equally at 4th and 2nd leaves emerged from the top) was the best model in this paper and referred as the water-nitrogen coupling model, which could control the number of rice tillering, improve the rate of tiller panicle, guarantee the appropriate LAI and grain-leaf ratio at filling stage, increase the efficient leaf area ratio and population light transmission rate, and thus improving photosynthetic capacity at filling stage and the accumulation of photosynthetic products, and on the premise of effective panicle and seed-setting rate, the yield increased significantly. However, the other combinations of irrigation regimes and N application managements resulted in weakened advantages, or even resulted in the negative effects of water-nitrogen interaction and decreased of grain yield. Especially, either W3 treatment or high N application ratio of later stage (N4 treatment) resulted in yield reduction, and aggravated negative effect of water-nitrogen interaction. According to yield performance under W3, the proper decrement of panicle N-fertilize ratios could ease the negative effect of water-nitrogen interaction to some extents, and suitable panicle N-fertilizer should account for 20%-40%. Under W1, however, on the basis of 40%-60% base tillering N fertilizer, panicle N-fertilizer should account for 40%-60%. Correlation analysis indicated that there existed significantly or highly significantly positive correlations (r=0.589*-0.978**) between index of population quality and yield under water-nitrogen interaction, especially had positive correlations between yield and increasing amount of dry matter at filling stage and decrement of population light transmission rate from full-heading stage to 20 days full-heading stage.【Conclusion】The results suggest that rice plant growth could be improved and high grain yield could be achieved through integrating and optimizing water-fertilizer regulating techniques in rice production. W2 and suitable N3 combined application was considered to be optimum under the experimental conditions, optimum N management pattern was also N3 for W1. However, under W3 treatment, the ratio of panicle N-fertilizer application should be decreased, and N management patterns (50% basal, 30% tillering, and 20% panicle N-fertilizer at 4th leaves emerged from the top) were considered to be optimum for W3.