A comprehensive analysis of a series of metabolites that produced by the organisms in the life processes will help to reveal the biological links between genotype and phenotype. Metabolomics is the method to analyze the metabolites by “omics”, which includes genomics, transcriptomics, and proteomics. To understand the biological response of plants to environmental stimuli or genetic interference, the application of the advanced analytical techniques combined with subsequent generation sequencing and the statistical methods were made for the extraction of information, data interpretation, identification and quantification of cellular metabolites. With the accumulation of high-throughput sequence information, emerged newly metabolomics database can be used not only to store, manage and analyze metabolomics data, but also to provide the chemical structure, physical and chemical properties and pharmacological properties, spectroscopy, experimental process, as well as to interpret the biological function of the metabolites. This paper described briefly several rather special metabolomics database resources (such as KEGG, METLIN, TOMATOMA, ReSpect, ECOMICS, PRIMe). Today, the effect of abiotic stress on plant metabolic spectrum is a focal point of metabolomics research. We reviewed the latest progress on plant responses to multiple environmental stresses such as hypoxia, UV-B radiation, high temperature, strong freezing, light and temperature environment, drought, and lack of sulfur and nitrogen. Future research will be mainly focus on the “key” variable site that decides the single biosynthetic step. This strategy can be used to identify genes and specific ways of crops and medicinal plants. Effects of mutations and transgenic event on plant metabolites have been attracted extensive attention. Glucosinolate (GSL) metabolic pathways provide a useful model system for the research of genome structure of the quantitative characters, while the genome-wide association studies (GWAS) fast become the preferred method for analyzing complex quantitative traits. Transgenic plant overexpressing will lead to the alteration of metabolites and proteins that could help to enhance the multiple stress tolerances. However, the safety assessment of genetically modified crops for food will still be questioned. Instead of relying only on a single analytical technique, identification of the metabolites needs integrating multiple “omics”. In the long-term evolutionary process, the plants will synthesize new metabolites due to the mutation from the metabolic pathways of the plants. The previous study showed genes that encoded the biosynthetic enzymes in plant metabolic pathways were co-regulated through the transcription factors. Thus, the chemical diversity of the plants is closely related to evolution of the transcription factors.
A comprehensive analysis of a series of metabolites that produced by the organisms in the life processes will help to reveal the biological links between genotype and phenotype. Metabolomics is the method to analyze the metabolites by ‘omics’, which includes genomics, transcriptomics, and proteomics. To understand the biological response of plants to environmental stimuli or genetic interference, the application of the advanced analytical techniques combined with next generation sequencing and the statistical methods were made for the extraction of information, data interpretation, identification and quantification of cellular metabolites. With the accumulation of high-throughput sequence information, emerged newly metabolomics database can be used not only to store, manage and analyze metabolomics data, but also to provide the chemical structure, physical and chemical properties and pharmacological properties, spectroscopy, experimental process, as well as to interpret the biological function of the metabolites. This paper describes briefly a few rather special

Chalcone synthase (CHS) is the first enzyme in the flavonoid biosynthetic pathway in plants. In order to clone CHS gene in Cryptomeria plants, the amplified consensus genetic markers (ACGM) were designed and combined with RACE technique, three CHS genes in Cryptomeria japonica var. sinensis，C. japonica cv. Araucarioides and C. japonica were obtained, respectively. Sequence analysis showed that the coding sequence (CDS) of each CHS gene was 1176 bp in length and encoded 391 amino acids. The GFGPG tag of CHS conservative active sites were found in their proteins. Three CHS proteins showed 99% homology with each other, but more than 79% homology with other plants, which indicated that CHS were relatively conserved during evolution.

In past two decades, abundant researches have been published on transformation, regeneration, and genetic enhancement of cotton, especially Gossypium hirsutum. Generally, genes conferring agronomic advantages have been introduced into plants via Agrobacteriummediation or particle bombardment, and then the transgenic plants are regenerated through somatic embryogenesis from callus. However, the embryogenic callusbased regeneration is difficult and timeconsuming in cotton. Therefore, it is necessary to establish an effective system for the Agrobacteriummediated genetic transformation of shoot tip in cotton. Triacylglycerols (TAG) are a heterogeneous group of molecules with a glycerol backbone and three fatty acids attached by ester bones.
Diacylglycerol acyltransferase (DGAT; EC3.2.1.20) catalyzes the last step of TAG biosynthesis and it is the only key enzyme evolved in. DGAT1 and DGAT2, as two types of DGAT in eukaryotes, belong to different gene families. And previous studies have reported that the expression of DGAT1 increased seed oil content and mass. In order to get new cotton germplasm with high oil content, we used the shoot tips of ‘Zhongmiansuo 49’ as explants and introduced an improved vector carrying a selection marker HptⅡ gene and a target VgDGAT1a gene into cotton via Agrobacteriummediated transformation. This improved Agrobacteriummediated transformation and regeneration system were established by optimizing different parameters such as preculture period of seeds, concentration of Agrobacterium in solution, immersing time, cocultivation period and components of MSB ［Murashige and Skoog (MS) medium + vitamins of Gamborg’s (B5) medium］. Cotton seeds (Gossypium hirsutum L. cv. ‘Zhongmiansuo 49’) were decorticated manually and surface sterilized in 0.1% HgCl for 8 min, followed by rinsing with sterile distilled water for five times. The cotyledons were removed from 3dayold to 5dayold in vitro germinated seedlings and the shoot tips were cut in lengthwise to decrease the damage of meristematic cells. The strain EHA105 was grown overnight on a shaker at 200 r/min and at 28 ℃ until the A600 value of bacterial concentration reached 0.60.9. The suspension cells were centrifuged at 5000 r/min for 10 min and the pellets were resuspended in an equivalent volume of liquid cocultivation medium ［MSB + 200 μmol/L acetosyringone （AS）］. The treated explants were immediately immersed into prepared Agrobacterium suspension containing pCAMBIA1301, a binary vector carrying the VgDGAT1a gene, for 4060 minutes. The tips were blotted dry on sterile filter paper and transferred into the cocultivation solid medium at 28 ℃ in dark. After cocultivation for 1 day, shoot tips were transferred into root induction medium containing MSB, 1 g/L activated charcoal, 400 mg/L cephalothin (Cef) and solidified with 0.2% (W/V) phytagel, then grew in a growth chamber with stringent light, temperature and humidity control. In the following 34 weeks, 5leafplants were transferred to plastic pots containing soil matrixes (1∶1 of turf and vermiculite). Besides the optimized genetic transformation protocol of cotton shoot tip as stated above, it is found that 50 mg/L hygromycin (Hyg) could accurately distinguish the resistant plants. In addition, reduplicated selections improved accuracy. In conclusion, this study describes an optimized transformation protocol for shoot tip of ‘Zhongmiansuo 49’ with an A. tumefaciens strain EHA105 harboring DGAT1 gene, and proves that it is an efficient and economical method to obtain transgenic plants based on the results of different and important parameters influencing the transformation efficiency.

Determination of chlorophyll concentration has significance for photosynthesis and stress physiology since it is the main photosynthetic pigment in higher plant leaves. Previous researches have shown high correlation between leaf chlorophyll concentration and SPAD values, obtained by SPAD-502Plus chlorophyll meter. Path analysis also has been used for analyzing the direct effect and indirect effect among various independent variables and dependent variable. There exist large differences in leaf characteristics among three species of Berberidaceae i.e Mahonia bealei (Fort.) Carr., Mahonia fortunei (Lindl.) Fedde, and Nandina domestica. In order to study the correlation of SPAD values and chlorophyll concentration in these three species, the chlorophyll concentration was measured by SPAD-502Plus chlorophyll meter and ultraviolet spectrophotometer. The chlorophyll concentration was expressed as per unit leaf area or per unit leaf fresh mass respectively; a parameter SLW (ratio of leaf fresh mass and leaf area), was introduced to describe the characteristic of leaf thickness. The effects of both the leaf apparent characteristics and chlorophyll parameters on SPAD values were further investigated by path analysis. Moreover, the empirical relationships between the SPAD values and the concentration of chlorophyll a (Chla), chlorophyll b (Chlb) and total chlorophyll (TChl) were determined by a linear function, a second-order polynomials function, an exponential function and a power function to calculate the R2 decision value based optimal fitting curve. For all three species, when chlorophyll concentration was expressed as per unit leaf area, it showed significant correlation with SPAD values. The SPAD values showed higher correlation with the concentration of Chla than Chlb in M. bealei and M. fortunei, and the result was
opposite in N. domestica. The positive correlation between parameter SLW and SPAD values was found among the three species and was significant in M. bealei and M. fortunei, which indicated that thick leaves contained higher chlorophyll concentration. Furthermore, SLW showed higher positive correlation with carotenoid (Car) than Chla, Chlb, TChl in all three species, which could be related to the stress resistance of Car; The path analysis led to two fold discoveries: 1) The concentration of Chla had the greatest impact on SPAD values; 2) The SPAD-502Plus chlorophyll meter could effectively eliminate the impact factors (e.g., leaf thickness). A non-linear relationship between SPAD values and chlorophyll concentration was indicated by the empirical equations; nevertheless, the optimal equations of Chla, Chlb, and TChl in M. bealei. were exponential function, while the power function of Chla, TChl and the second-order polynomials function of Chlb in M. fortunei and N. domestica were observed. The correlation between SPAD values and chlorophyll concentration in three different species of Berberidaceae indicats that the SPAD values can accurately reflect the chlorophyll concentration in higher plant leaves and the SPAD-502Plus chlorophyll meter can effectively eliminate the impact factors, so we propose that the SPAD-502Plus chlorophyll meter can be referred as a fast and nondestructive method to evaluate the chlorophyll concentration, and SPAD calibration curves
shall commonly be parameterized as non-linear relationship.

The structure of rice spikelet, an important determinant of yield and rice quality, is a hot topic in rice research areas. The structure of spikelet is somewhat complex, and its development is influenced by multiple factors, including genetic factors and environmental factors. Two major technical routes in studying molecular mechanism of the structure of spikelet are used: one is homologous sequencing of the dicotyledonous plant to screen the rice cDNA library and obtain the related genes, and the other is creating some spikelet character mutants with locating and cloning the mutant genes to grasp the genes function included. In order to further study rice spikelet development and its mechanism of regulation, we used a spikelet degradation mutant (spd-hp73), which was obtained from the offspring of hp73 mutated by 60Co-γ radiation. In this study, the characteristic and genetic stability of rice spikelet of spd-hp73 mutant in different environments were investigated. And three crosses were made between the spikelet degradation mutant (female parent) and the wild-types Zhe7954，9311，Minghui63 (male parents) to investigate the inheritance pattern of the mutation. And 519 SSR molecular markers were used for mutant genetic analysis. The mapping populations were derived from a cross between spd-hp73 and Zhe7954.
Compared with the wild-type (hp73), the spd-hp73 mutant showed several abnormal agronomic characteristics, including weak growth, early flowering, low germination, scrubby plant, few tillers, thin stems, short leaves and clear closure of spikelet. Furthermore, the branches were twisted at different degrees, and the secondary branch was longer than hp73. Moreover, spd-hp73 exhibited abnormal inflorescence architecture, including the abundant degradation of spikelet, significant reduction of the grains per spikelet, low grain density and decreased seed setting rate. The traits of mutant were expressed stably in different growth environments. Spikelet degradation mutant resulted in a remarkable negative effect on rice normal growth and reproductive growth. Genetic analysis indicated that the special phenotype was controlled by a single recessive gene, named spd-hp73. The mutation site was preliminarily mapped to the long arm of chromosome 4 between SSR markers RM471 and RM273 with 12.2 cM and 9.4 cM intervals, respectively. These results of preliminary genetic analysis lay a foundation for fine mapping and cloning of spikelet degradation gene. Probably, spd-hp73 plays an important role in further understanding the regulation mechanism of entire spikelet development.
In order to further study of rice spikelet development and its mechanism of regulation, we used a spikelet degradation mutant (spd-hp73), which was obtained from the offspring of hp73 mutated by 60Coγ radiation. In this study, the characteristic and genetic stability of rice spikelet in different environments of spd-hp73 mutant were investigated. And three crosses were made between the spikelet degradation mutant (female parent) and the wild-type Zhe7954，9311，Minghui63 (male parents) to investigate the inheritance pattern of the mutation. And 519 SSR molecular markers were used for mutant genetic analysis. The mapping populations were derived from a cross between spd-hp73 and Zhe7954. Compared with the wild-type (hp73), the spd-hp73 mutant showed several abnormal agronomic characteristics, including weak growth, early flowering, low germination, scrubby plant, few tillers, thin stems, short leaves and clear closure of spikelet. Furthermore, the branches were twisted as different degrees, and the secondary branch was longer than the hp73. Moreover, the spd-hp73 exhibited abnormal inflorescence architecture, including the abundant degradation of spikelet, significant reduction of the grains per spikelet, low grain density and decreased seed setting rate. The traits of mutant were expressed stably in different growth environments. Spi

Rice sheath blight caused by Rhizoctonia solani Kühn is one of the most important diseases on cultivated rice worldwide. Unlike most other fungal pathogens, R. solani forms heterokaryotic vegetative mycelia with multiple nuclei per hyphal cell and is unable to produce haploid asexual spores under normal conditions. These characteristics of R. solani may make it difficult to perform genetic transformation and functional analysis of genes. Production of large amount of protoplasts from this fungus is a prerequisite for the studies of molecular genetics, such as protoplast fusion and fungal transformation. Previously, some lytic enzymes and conditions for releasing R. solani protoplasts have been tested and optimized and several protocols for the preparation and regeneration of protoplasts from R. solani mycelium have been developed by some researchers. However, the efficiency of R. solani protoplasts releasing by these protocols is sometimes unstable due to different strains of R. solani or experimental conditions. Therefore, it is necessary to develop an efficient method for preparing protoplasts of rice sheath blight fungus. The objectives of the present study were to evaluate various cell wall degradation enzymes and their combinations for releasing protoplasts from R. solani mycelium, and to develop an efficient protocol for yielding protoplasts. Using 07 mol/L NaCl as stabilizer solution, seven different cell wall degradation enzymes, including Glucanex, lywallzyme, cellulase-R-10, macerozyme-R-10, snailase, driselase and lysing enzyme, and their combinations were evaluated for releasing protoplasts from R. solani GD-118 mycelium which was harvested from potato dextrose liquid medium cultured at 28 ℃ for 36 h. The number of released protoplasts was counted by using haemocytometer under microscopy. The optimal concentration of lytic enzymes for the generation of protoplasts was determinated and the conditions to obtain and regenerate protoplasts of the fungus were also optimized. Among the seven tested lytic enzymes, Glucanex was the most suitable enzyme for the digestion of R. solani GD-118 cell wall. The protoplast yield in the treatment with 20 mg/mL Glucanex for 4 h was 237×104 cell/mL (e.g. 1185×104 cells per gram mycelium). Moreover, the results showed that the optimal mixture enzymes of 15 mg/mL Glucanex and 10 mg/mL lywallzyme were effective in releasing protoplasts with the production of 309×107 protoplasts from per gram R. solani GD-118 mycelium, and the obtained 58% protoplasts could be regenerated. In addition, the combination had similar effects on digesting cell wall of different strains of rice sheath blight fungus. Taken together, the mixture lytic enzymes of 15 mg/mL Glucanex and 10 mg/mL lywallzyme can effectively digest the cell wall of rice sheath blight fungus and produce abundant protoplasts.

With the increasing demand for vegetables, continuous vegetable cropping was more popular and common in China, which will result in seriously continuous cropping obstacle in fields. Imbalance of soil microbial community structure, decline in bio-diversity, reduction in quantity of beneficial microorganism and accumulation of pathogenic microorganism were major factors of continuous cropping obstacles. Soil microorganism plays an important role in maintaining balance, health and high quality of soil ecosystem. Changing of microbial community structure directly influenced soil function, thereby produced negative effect on plant growth. Using artificial regulation measures such as adjusting microbial community structure and increasing bio-diversity, was an effective method to overcome continuous cropping obstacles. Calcium cyanamide and dicyandiamide (DCD) were cyanamide fertilizers, which had significant inhibition effect on harmful organisms, especially on pathogenic fungi in soil. However, effects of cyanamide fertilizers on microbial community structure and bio-diversity of continuous cropping soil were unclear. The aim of this research was to study the effects of calcium cyanamide and dicyandiamide application on microbial community structure in continuous vegetable cropping soil. Results of the study might reveal the soil micro-ecological mechanism of calcium cyanamide and dicyandiamide to control eggplant verticillium wilt, and provide theoretical foundation and technical guidance for cyanamide fertilizers application in agricultural production.
The experimental soil was silt paddy soil and was collected from farmland in Quzhou, Zhejiang Province. In the farmland, the planting years of eggplant were more than four years, and verticillium wilt of eggplant was severe and out of control in the last year. Six treatments were designed in experiments: 1) control, 2) dicyandiamide, 3) calcium cyanamide (lime nitrogen), 4) straw, 5) dicyandiamide + straw, 6) calcium cyanamide + straw. Soils of different treatments were incubated at 25 ℃. Through conventional artificial culture method, quantities of bacteria, fungi and actinomycetes were recorded. Community structure and bio-diversity of bacteria and fungi were analyzed by PCR-DGGE.
Results showed that, calcium cyanamide application effectively increased the number of bacteria, decreased the amount of fungi, improved the ratio of bacteria to fungi and actinomycetes to fungi in the continuous cropping soil. But dicyandiamide showed no distinct influence on the number of bacteria. Dicyandiamide and dicyandiamide plus rice straw application increased the ratio of bacteria to fungi with the high value of 2.48 and 9.96 times in the 30 days. Calcium cyanamide and calcium cyanamide plus rice straw application obviously increased the ratio of bacteria to fungi with the high value 8.42 and 58.39 times in the 30 days. So, application with calcium cyanamide in the continuous cropping soil could change the microbial community structure, restore the imbalance microbial community structure into a healthy status for vegetable growth by changing the dominant microbial community structure from fungi to bacteria, and then controled the soil-borne disease in the continuous vegetables cropping soil. The results of PCR-DGGE also indicated that, cyanamide fertilizer application altered the community structure of bacteria and fungi, induced the appearance of new species and biotic population, and inhibited some other population. In the continuous vegetables cropping soil, the diversity index and richness of bacteria were decreased with the calcium cyanamide application in the early stage (8 days after application), but the bacteria community diversity and richness were increased in the later stage (20 days after application). Dicyandiamide application obviously increased the diversity index and richness of bacteria than calcium cyanamide application. The calciu

Gynoecy plays an important role in cucumber (Cucumis sativus L.) heterosis breeding and identification of the markers linked to this character will facilitate selection of gynoecious cucumber line in breeding program. Traditional selection for cucumber cultivars with gynoecious line has required evaluation in complicated environments over several years, which is long period, time and labor consuming.
Molecular markers offer a faster and more accurate way for breeding, as selection can be based on genotype rather than phenotype. The use of molecular markers for indirect selection of important agronomic characters, or marker-assisted selection (MAS) can improve the efficiency of traditional breeding. Many studies developed a lot of SSR markers, which had greatly facilitated MAS in cucumber breeding. Now some studies showed that some markers were connected with gynoecious gene but the distances were not compact, so few were availably applied to breeding. The aim of this study was genetic analysis of gynoecy and identification of molecular marker associated with gynoecious gene using gynoecious line, monoecious line, and SSR marker. The genetic analysis of cucumber gynoecious was evaluated with a gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2 and their F1, F2, BC1P1, BC1P2 populations in the present study. Total DNA of parents and F2 were isolated from freeze-dried leaf tissue by the CTAB method. SSR markers were analyzed with
gynoecious line 240-1-2-2-3-1, monoecious line 3-5-1-3-2-1-1-1-1-2, F2, and 699 pairs of SSR primers. SSR analysis was performed with the primers. PCR was performed in 20 μL reaction containing 2 μL genomic DNA (20 ng), 10 pmol/μL primers 0.4 μL, respectively, 2.5 mmol/L Mg2+ 2 μL，2 mmol/L dNTP 1 μL，5 U/μL Taq DNA polymerase enzyme 0.1 μL，10×buffer 2 μL and double distilled water. The amplification profiles were 5 min at 94 ℃, followed by 35 cycles of 30 s at 94 ℃; 1 min at 55 ℃, 1 min at 72 ℃; then 10 min at 72 ℃. After amplification, the PCR products were mixed with loading buffer (2.5 mg/mL bromophenol blue, 2.5 mg/mL diphenylamine blue, 10 mmol/L EDTA, 95% (V/V) formamide), denatured for 5 min at 94 ℃ and put on ice for 5 min. The denatured PCR products were separated on 6% (W/V) denaturing polyacrylamide gel at 100 W power and visualized by silver straining. Polymorphic fragments of primers were cloned and sequenced. Linkage analysis used the software of Mapmaker V3.0. During analysing the separated rate of F1 and F2, the results showed that the gynoecy in 240-1-2-2-3-1 was controlled by oligogene with some background genes modified. Inheritance of gynoecy was accord with the additive-dominant-epistatic model. From 699 pairs of SSR primer，two pair of stable SSR markers (CSWCT25 and SSR18956)，331 bp and 145 bp in bands size were obtained respectively during PCR products of two SSR markers cloned and sequenced, and linkage analysis indicated that its genetic distance to the gynoecious loci was 7.7 cM and 6.8 cM, respectively.
Two SSR markers are tightly linked to gynoecious loci on the chromosome 6. In sum, knowledge of location of gynoecious gene in cucumber and its
related traits in crosses will be helpful to the design of more effective selection schemes to develop gynoecious cucumber genotypes. Progress in breeding gynoecious lines is still slow because of the complex inheritance of this character. However, gynoecious cultivars and the markers identified in this study can contribute to improving gynoecious line. Two SSR markers could be used effectively for molecular marker-assisted selection in breeding programs to develop cucumber gynoecious line breeding.

Soil respiration is the primary way by which CO2 absorbed by terrestrial plants returns to the atmosphere. And it may have distinctly dynamic patterns at different temporal scales since it is affected by diverse abiotic and biotic factors. Increasing deposition of nitrogen from the traditional cultivation of sympodial bamboos may lead to the sequestration of carbon in vegetation and soil. And the rising temperature and water content may increase the flux of CO2 from the soil, but the response of the ecosystem to simultaneous changes in all of these factors is still unknown. Meanwhile, to provide abundant supply of bamboo timber, afforestation of bamboo species such as Oxytenanthera braunii Pilger ap. Engler, Dendrocalamus brandisii Kurz and D. giganteus Munro is encouraged by the government but without scientific directions. And chemical fertilizers are usually applied into fields unscientifically and blindly in the villages of southwest China. Subsequently, what will happen to the soil structure and how to balance soil nutrient environment in the situation of chemical fertilizer abuse?
In the context of climate change, the amount of nitrogen allocated to the soil is predicted to increase with the productivity of terrestrial ecosystem, and may alter soil carbon storage capacities. To provide the proof of soil respiration responding to the nitrogen input for sympodial bamboo afforestation at the beginning period, we set up four nitrogen fertilization (CO(NH2)2) levels in mid-high mountain of southeast China, i.e. N content of 0, 40, 80, 160 kg/hm2
(expressed on N0,
N40, N80, N160, respectively), using the two-year old stump of wine bamboo which were planted every five meters. The soil respiration rate is measured by using trenching method and infrared gas analyzer. The responding mechanism is discussed through analyzing the change of soil temperature at 10 cm depth (T10), as well as changes of soil water-soluble organic carbon content (WSOC) and soil water content (SW).
Results showed that soil respiration rate was quite different between rainy and dry seasons. The soil respiration rate increased at the end of April or in the beginning of May when the rainy season arrived. Its wave crest arrived in July, Aug. and Sept., and then the rate decreased along with the dry season in Nov., Dec., Feb. and Mar., then the trough of soil respiration rate appeared. The variation rule of T10, WSOC and SW was similar as this way. Exponential function could be used to describe the relationship between T10 and respiration rate. Meanwhile, WSOC and SW showed a linear relationship with the respiration rate respectively, and the regression test
indicted that it was significant. And the temperature sensitivity value Q10 of a whole year was 2.45-2.78 nearby. In rainy season, Q10 decreased to 1.66-1.89, which indicated that the sensitivity of respiration rate responding to temperature decreased. On the contrary, Q10 ascended to 4.85-9.54 in dry season. The yearly data of WSOC were unstable, and the nitrogen input could not enhance T10 and SW, but N80 and N160 could increase WSOC relatively. The changes of SW and T10 explained 96.10%, 94.30%, 94.48% and 92.99% of the variation of soil respiration rate in the treatment of N0, N40, N80 and N160, which contributed most of the information. The main factor affecting the soil respiration in rainy and dry seasons was quite different, which was SW and T10 respectively.
As a consequence, the increase in ecosystem productivity may lead to an increase in carbon turnover in the soil, via an increase in the amount of biomass. But its process and mechanism involving different carbon pools are very complex, and to measure the soil respiration rate alone can not totally reflect the whole change of carbon cycle. Experiments of further

Aroma is an important factor to assess the quality of tea. The same variety of tea plants in different regions may generate different types of aroma. Thus, choosing the appropriate region is the basic principle to gain high quality of tea. Earlier studies showed that several soil elements play an important role in taste of tea. However, few studies have been reported about their effects on tea aroma. The goal of the present study is to further investigate the relationship between soil conditions and tea aroma, in order to better manage the tea plantation. For this purpose, the soil of tea garden in three different producing regions and corresponding tea aroma were analyzed. The three regions were named region 1 （Wufeng, Changnan）
, region 2 (Anfeng, Wuyi) and region 3 (Panban, Yuhang) respectively. Potash fertilizer test of three levels, nonpotassium chloride (control), 200 kg/hm2 potassium chloride and 400 kg/hm2 potassium chloride were carried out in the experimental base of Zhejiang University. Tea volatile compounds were collected by headspace solidphase microextraction and were analyzed by gas chromatographymass spectrometry. Volatile compounds were identified on mass spectra library and retention index. The quality of tea was also judged by sensory evaluation. Soil effective mineral elements were extracted by Mehlich 3 extractant and measured by ICPOES. All of the data were analyzed by oneway ANOVA followed by Tukey’s test. Sensory evaluations showed that tea aroma characteristics of region 1 and region 2 were much better than that of region 3. In addition, results of volatile compound analysis showed that in region 3, the total contents of alkenes and alcohols in tea were as little as 21.09%, of which favorable alkenes and alcohols were only 17.46%. Nevertheless, its content of grass compounds was as high as 8.53%. Compared to region 3, the total contents of alkenes and alcohols in region 1 and region 2 increased by 154%, 172% respectively. Meanwhile, the favorable alkenes and alcohols increased by 101%, 130% respectively, and their contents of grass compounds were both lower than 3%. Ten volatile compounds with favorable odour
(e.g., nonanal, benzyl alcohol) were detected merely in region 1 and region 2. The result illustrated that the content of favorable alkenes and alcohols, as well as grass compounds had an important influence on aroma of tea. On the other hand, results of soil element analysis showed that it exhibited significant increase of effective potassium, magnesium and phosphorus in region 1 (120%, 159% and 30% respectively) and region 2 (170%, 59% and 70% respectively), compared to region 3. Furthermore, potash fertilizer experiment indicated that potassium could improve aroma of tea. As shown in ANOVA analysis, alkenes, alcohols, ketones and aldehydes increased significantly in potash fertilizer treatments compared to the controls （CK）. With increasing level of potash fertilizer, Favorable alkenes and alcohols increased by 25% and 48% respectively, while grass compounds decreased by 31% and 40% respectively. In conclusion, a complementary supply of magnesium, phosphorus and especially potassium for poor tea soil should be taken into consideration, based on current tea plantations.

Substrate filler is one of the important equipments for mechanization growing seedlings. Mechanized breeding has been developed as a new horticulture technology in recent ten years worldwide. It has become a professional commercial seedling production way in many countries, particularly in Europe and America. Development of substrate filler machine and seedling production line can promote the seedling production in protected horticulture. With the rapid development of modern horticultural production, the demand for substrate filler is increasing dramatically. However, the composition of substrate filler is very complicated in structure. It is difficult to develop a single type of substrate filler to meet the demand of all crop growth. Furthermore, automatic feeding device is complex and expensive in the fierce competition in the growing market. Therefore, people are looking for more convenient and effective seedling medium automatic feeding devices, which can not only fill substrate in plugs, but also mix matrix uniformity according to various specifications.
Now, the design, performance testing and filled result of TLZ-400 type substrate filler
were reported. This filler was composed of material tank, lifting device, matrix tray conveying agency, vibration, rotating scraper, bodies brush, and electrical control components. The working principle is that the mixing substrate is transported to plug over and fall into the tray on a conveyor belt after promotion agencies raise, followed by scraper shaving, vibration shaking, and brush cleaning, so as to achieve the purpose of smooth and uniform mixing of substrate in plugs. The purpose was to investigate and confirm the main technical parameters of TLZ-400 type substrate filler.The technical characteristic parameters，productivity, and power consumption of TLZ-400 type substrate filler were measured by using selected perlite (50%), organic fertilizer and black earth (30%), and sawdust (20%) as mixed substrate. Substrate uniformity, sample standard deviation, and sample coefficient variation were also analyzed by using substrate sampling methods of 18-36 holes in plugs.
Results showed that supporting power of this substrate filler was 1180 W and the production capacity ranged from 50 to 380 tray/h. When the vibration status was off or on, the total standard deviation was 1.485 or 1.247, and the total coefficient variation was 7.94% or 6.70%, respectively. We conclude that the TLZ-400 type substrate filler is suitable for matrix filler, vibration, cleaning, and recycling. It is ready for seeder sowing after the substrate is uniformly filled in nursery plug.

The pollution of heavy metal is increasingly concerned, especially the soil heavy metals pollution around the contaminated sites. Many researchers have reported a significant increase of heavy metal in the surface soil. However, few reports have been found in the literature about soil heavy metal spatial distribution and source identification around contaminated sites. Therefore, this study has the aim of elucidating the spatial distribution and source identification of soil heavy metals in croplands near two contaminated sites. Our specific objectives were to examine the spatial dependency and the variation mechanism of heavy metals in soils, to map the spatial distribution and risk assessment of soil heavy metals, and to identify the main sources of soil heavy metal pollution.
The surface soil (0-20 cm) and subsurface soil (20-40 cm) were collected from nearby fields of a battery plant (F1) and a thermal power plant (F2) in the north of Zhejiang Province. The concentrations of soil mercury (Hg), cadmium (Cd), lead (Pb), zinc (Zn), copper (Cu), and nickel (Ni) in these two typical contaminated sites were determined and the spatial distribution patterns and pollution source of the heavy metals were analyzed using Geographic Information System (GIS) and geostatistical methods. Based on the type II limit of “Environmental Quality Standard for Soils” (GB15618—1995), the single factor method and Nemerow index method were used to assess the comprehensive pollution risks of the soil heavy metals around the contaminated sites. Inverse distance weighted interpolation (IDW) was used to simulate the pollution risk and spatial distribution and the source identification of these heavy metal pollution.
Results showed there was a serious soil heavy metal pollution problem around contaminated sites, with Cd concentrations in 80.77% of the surface soil samples exceeded the type II limit (Cd>0.3 mg/kg) of GB15618—1995. Hg concentrations in 3.85% of the surface soil samples exceeded the type II limit. Nemerow index method assessment results showed that there were 25 soil samples exceeded the standard, with overproof rate of 48.08%. Based on the Technical Specification for Soil Environmental Monitoring, the maximum index value was 3.13, which exhibited heavy pollution. Particularly in nearby contaminated site F1, soil Cd and Pb pollution were serious problems in the soils, with Cd concentration in 91% of the surface soil samples exceeded the type II limit (Cd>0.3 mg/kg) of GB15618—1995, and the maximum surface soil Cd concentration was 3.3 mg/kg. The soil Pb pollution was also the potential problem even then its concentration was not exceeded the type II limit. The maximum surface soil Pb concentration was 123.67 mg/kg, and the mean surface soil Pb concentration was 58.69 mg/kg. While the surface soil Cd and Pb concentrations were significantly higher than the subsurface soil. At the nearby contaminated site F2, soil Cd and Hg pollution were serious problems in the soils. Cd concentrations in 63% of the surface soil samples, and Hg concentrations in 11% of the surface soil samples were higher than the type II limit (Cd>0.3 mg/kg, Hg>0.3 mg/kg) of GB15618—1995. The maximum surface soil Cd concentration was 0.71 mg/kg. The mean surface soil Hg concentration was 0.214 mg/kg. Cd and Pb concentrations in the surface soil samples from F1 had similar spatial distribution patterns and the concentrations reduced with farther distance away from F1, and they were significantly higher than those in the corresponding subsurface soils. These results indicated that pollution of soil heavy metals (Cd and Pb) was mainly caused by external pollution, and F1 was the most probable pollution source. The battery plant can discharge Cd and Pb into environment. At the contaminated site F2, the heavy metal concentrations in some soil samples were higher than the type II lim

Presently, obesity is not only becoming a critical medical problem worldwide but also will cause lots of obesity-related chronic diseases as well, like hypertension, cardiovascular diseases, diabetes and so on. It becomes more and more important for current scientists to explore the effective pathway to solve the puzzle. Many researchers from different countries put much more emphasis on the anti-obesity effect of natural products. There are many studies showing that the extracts of certain parts of varied kinds of natural plants are able to inhibit the obesity through the influences on lipids metabolism, adipocytes secretion and show prevention effects on obesity through animal tests. Yellow capsicum includes functional component capsaicin which has multifunctional pharmaceutical effects. In the present study, the anti-obesity action of yellow capsicum extract (YCE) was investigated through their intervention effect on lipids metabolism in C57BL/6J mice. The total thirty-two mice were divided into four groups, including control group, obesity model group and YCE treatment groups at doses of 100 and 25 mg/kg body mass. The mice in control group were fed with normal diet and those in other groups were fed with hyperlipidic diet and YCE at different doses once a day for 40 consecutive days. All mice were weighed every five days and their body mass were recorded. In the end, the mice were killed and dissected, body fat percentage, contents of total cholesterol (TC), triglyceride (TG), high-density lipoprotein (HDL), low-density lipoprotein (LDL) and leptin in serum, expression levels of TC, TG, aspartate aminotransferase (AST) and alanine aminotransferase (ALT) in the liver, were all detected and analyzed. Besides, the pancrelipase (PL) level of pancreatic tissue was also detected through ELISA method, as it is an important lipase in lipids metabolism.
The results showed that, after 40 days’ treatment through hyperlipidic diet, the obesity model group was successfully built up, as they were overweighed (P<0.01) and showed a higher mass of adipose tissue (P<0.05), TC (P<0.05), TG (P<0.01) contents than the normal diet control group. Compared with obesity model group, YCE exhibited an inhibitory effect on body mass to some extent (P<0.01), fat tissues, body fat percentage (P<0.01) and Lee’s index declined (P<0.05). The contents of TC, TG and LDL of the serum in YCE treatment groups were declined (P<0.01), while there was no difference in HDL level. The liver mass (P<0.01) and its TG contents (P<0.05) in YCE treatment groups were lower than that of obesity model group. Meanwhile, TC content exhibited no significant change, on the other hand, the AST and ALT levels in the liver were normal, it showed that the YCE treatment had no side-effect on liver tissues. The PL level in YCE treatment groups was decreased (P<0.05) and it indicated that YCE might act on PL to affect lipidolysis.
From the above tests we can conclude that YCE has a significant anti-obesity effect. The changes in blood and liver indices, like TC, TG and so on, reflects its intervention on lipids metabolism. Moreover, YCE has no side-effect on liver tissues. Also YCE has a potential inhibitory effect on Pancrelipase (PL), which can be a potential target for anti-obesity therapy. Our results provide a further research evidence and basis for YCE’s deep exploration and development. Presently, obesity is not only becoming a worldwide critical medical problem but also causes lots of obesity-related chronic diseases like hypertension, cardiovascular diseases, diabetes and so on. It becomes more and more important for current scientists, exploring the effective pathway to solve the puzzle. Many researchers from different countries are putting much more emphasis on the anti-obesity effect of natural products. There are many studies showing that the extracts of certain parts of various kinds of natura

Soy isoflavones (SIF) are a research focus due to their potential health effects. Some studies have shown that it could modulate immune function and prevention of chronic diseases, but until now it remains controversial. Thus, the aim of the present study is to investigate the effect of high-fat diet induced obesity on male immune parameters. Numerous studies have reported that immunomodulatory activity of intestinal mucosal barrier was related to intraepithelial lymphocytes (IEL) and goblet cells (GC). Leptin is secreted almost exclusively by adipocytes. Previous researches demonstrated that plasma leptin levels in ovariectomized rats was lower than that in estradiol-treated animals. Moreover, there was a tissue-specific regulation of the leptin receptor (OB-Rb), with less sensitivity to leptin in adipose tissue when the time of treatment or the dose of estradiol administered increased. The metabolites of soy isoflavones showed structural and functional similarities to estradiol. Available data indicated that soy isoflavones inhibited the leptin secretion, and thus influenced the expression of OB-Rb. The study was divided into two steps. In the first experiment, male Sprague-Dawley rats were fed with a high sugar and fat diet (HSF) or basal diet for eight weeks. Food intake and animal mass were recorded weekly. In the second experiment, 40 selected obese rats were randomly divided into four groups with 10 rats in each group and were fed with soy isoflavones (group Ⅰ 0 mg/kg, group Ⅱ 50 mg/kg, group Ⅲ 150 mg/kg and group IV 450 mg/kg) and 10 control rats (group V) were fed with basal diet for four weeks. Soy isoflavones was gavaged with 0.5% sodium carboxymethyl cellulose solutions respectively.
At the end of the second experiment, rats were slightly anesthetized with sodium pentobarbital ip (40 mg/kg), and intestinal was taken and fixed in 10% neutral buffered formalin, processed and trimmed, embedded in paraffin. IEL and GC in the villus epithelium were evaluated in these rats, as well as the OB-Rb expression by immunohistochemistry staining. In the first experiment, the body mass of the HSF-fed animals were significantly increased after eight weeks and the obesity module was succeeded. In the second experiment, the results showed that the structure of mucous epithelium, intraepithelial lymphocyte and the goblet cells were normal in the intestinal. The intraepithelial lymphocytes of rats were mainly small lymphocytes and located around the epithelial basement membrane. Goblet cells distributed in the intestinal mucous epithelial lamina, and OB-Rb positive cells distributed in the intestinal mucous epithelial lamina and lamina propria, respectively. Compared with 0 mg/kg-SIF group, there was a tendency that IEL in the villus epithelium moved inward intestinal entoderm in SIF-treated rats. Quantitative analysis of IEL and GC in the villus epithelium showed the number of GC in duodenum, jejunum, ileum, colon had a increasing tendency from the forepart to the back of intestinal.The number of IEL and GC in the epithelium in the 450 mg/kg-SIF group were increased significantly (P＜0.05), and there was significant difference in numbers of GC between the control and other groups. The changes of OB-Rb were relatively irregular in each part of the intestinal. The OB-Rb expression also increased significantly (P＜0.05) in intestinal mucous membrane in the 450 mg/kg-SIF group. IEL and GC may play a very important role in the local mucosa immunity. The low number of IEL and GC showed that obesity may impair the immune function of rats. The results showed that SIF can affect mucosal immunity by increasing the number of IEL and GC. The OB-Rb expression of SIF-treated rats was significantly increased compared with untreated ones. We deduced that SIF contains a number of biologically active components with potential immunomodulatory activity, and SIF could mod

Freeze-drying has been widely used in the production of pharmaceuticals, vaccines and high-valued foods. Foam freeze-drying is a new type of drying technology, in which the sample is foamed and frozen before drying. Compared with traditional freeze-drying method, the new method can reduce operation time and cost significantly. Some foreign researchers have used this method to produce heat-resistant vaccines which can endure higher temperature and remain effective after a longer storage in refrigerator. Up to now, little research about foam freeze-drying has been reported domestically. In this study,
newcastle disease vaccine was used as an example to illustrate the procedure of foam freeze-drying method and to test its capability of producing heat-resistant vaccine. The ingredient of the foam freeze drying (FFD) sample included w(trehalose)=10%, w(hydrolyzed gelatin)=5%, w(pluronic F-108)=3%,
w(arginine)=2%, 25 mmol/L phosphate buffer (pH=7.2). The sample was prepared through the following procedure: 1) One milliliter sample was placed on the plate of the freeze dryer and cooled to 15 ℃; 2) then the pressure was lowered to 5 Pa and foaming began; 3) after one hour the plate temperature was increased up to 31 ℃ at a velocity of 0.7 ℃/min and kept for another 48 hours. The samples made by foam freeze drying were analyzed with thermal analysis and aging test, and were compared with live attenuated samples and heat-resistant samples prepared by traditional freeze drying. The latter two kinds of sample were bought from market (Tianbang Biotechnology Corporation, Nanjing, China). Thermal analysis was done with a differential scanning calorimetry (DSC 200 F3 Maia, NETZSCH, Germany). The procedure was as follows: 1) the sample was cooled to -20 ℃ at a rate of 10 ℃/min and held for 3 minutes; 2) the sample was heated to 80 ℃ at the rate of 10 ℃/min and held for 3 minutes; 3) the sample was cooled to room temperature at the rate of 10 ℃/min. Aging test was carried out by placing the sample at 37 ℃ for 10 days and thereafter measuring its EID50. EID50 was determined by vaccination with chick embryo and Reed-Muench calculation method. The appearance of the FFD sample was like sponge, totally different from the flat biscuit-like sample made by traditional freeze drying method. The average glass transition temperature of FFD vaccine was 33.9 ℃, higher than 30.2 ℃ of live attenuated vaccine, but lower than 40.1 ℃ of heat-resistant vaccine. This means that the heat resistance ability of FFD vaccine is physically better than that of live attenuated vaccine. After aging test the titers decreased for FFD vaccine was 1.1, which was a little higher than 0.8 for heat-resistant vaccine. The latter result was in consistent with the former result. It is clear that FFD vaccine has its own superiority on heat resistance ability, although the formula and operation parameters still need to be improved. Considering its obvious economic prospect, it is worth to carry out further work on the study of FFD vaccine.