Polyphenolic Compound and the Degree of Browning in Processing Apple Varieties

Polyphenolic compound in processing apple （Malus domestica Borkh.） varieties and the relationship between polyphenol content and enzymatic browning were studied to provide reference for raw material selection and processing method optimization. The content of polyphenol compound in 10 processing apple varieties （4 cider and 6 juice varieties） were analyzed using the Folin-Ciocalteu method and HPLC. The degree of browning and the activities of polyphenol oxidase were also studied. The content and proportion of the polyphenol varied depending on the variety. Bitter varieties globally showed a higher polyphenol concentration than sweet or acid varieties. Proanthocyanidins, chlorogenic acid, （＋）-catechin, （-）-epicatechin were high-concentrated polyphenols in apple fruits. Phloridzin, the unique polyphenol of apple, was abundant in the bitter variety Frequin rouge fruit. Total polyphenols, proanthocyanidins, （＋）-catechin, and phloridzin had higher correlations with browning. The correlation was low between chlorogenic acid and browning. The polyphenolic profiles were correlated with the apple types. Cider apples contained more polyphenol than juice apple varieties. The content of flavan-3-ol has a close relationship with fruit browning.     

Overexpression of a Cytosolic Ascorbate Peroxidase Gene,OsAPX2, Increases Salt Tolerance in Transgenic Alfalfa

Alfalfa （Medicago sativa L.） is an important forage crop in the world and it is of great signiifcance for the improvement of its salt tolerance. To improve salt tolerance in alfalfa, a rice ascorbate peroxidase gene （OsAPX2） was introduced into alfalfa using Agrobacterium tumefaciens-mediated transformation with marker gene bar. The different T-DNA insertions in T1 transgenic alfalfa were identiifed by Southern hybridization. Three independent T2 transgenic lines were selected for stress analysis and the results showed that all of them were salt tolerant compared with wild-type plants. The transgenic plants had low levels of H2O2, malondialdehyde and relative electrical conductivity under salt and drought stresses. Moreover, the contents of chlorophyll and proline, and APX activity were high in transgenic plants under salt and drought stresses. Taken together, the overexpression of OsAPX2 enhances salt tolerance in alfalfa through scavenging reactive oxygen species.     

Genome-wide identification,molecular evolution,and expression divergence of the hexokinase gene family in apple

Hexokinase(HXK) is the first irreversible catalytic enzyme in the glycolytic pathway, which not only provides energy for plant growth and development but also serves as a signaling molecule in response to environmental changes. However, the evolutionary pattern of the HXK gene family in apple remains unknown. In this study, a total of nine HXK genes were identified in the Malus×domestica genome GDDH13 v1.1. The physiological and biochemical properties, exonintron structures, conserved motifs, and cis-elements of the MdHXK genes were determined. Predicted subcellular localization indicated that the MdHXK genes were mainly distributed in the mitochondria, cytoplasm, and nucleus. Gene duplication revealed that whole-genome duplication(WGD) and segmental duplication played vital roles in MdHXK gene family expansion. The ω values of pairwise MdHXK genes indicated that this family was subjected to strong purifying selection during apple domestication. Additionally, five subfamilies were classified, and recent/old duplication events were identified based on phylogenetic tree analysis. Different evolutionary rates were estimated among the various HXK subfamilies. Moreover, divergent expression patterns of the Md HXK genes in four source-sink tissues and at five different apple fruit developmental stages indicated that they play vital roles in apple fruit development and sugar accumulation. Our study provides a theoretical basis for future elucidation of the biological functions of the MdHXK genes during apple fruit development.     

Transformation of TrxS Gene into Barley by Particle Bombardment

The production of malting barley in China can＇t meet the demand of beer industries because of poor quality and it becomes a bottleneck problem in beer manufacture industry. In this paper, TrxS gene cloned from Phalaris coerulescens was transferred into barley cultivar Yupi 1 （YP1） via biolistic bombardment. 1206 immature embryos were bombarded and seven transgenic plants carrying TrxS gene were confirmed by PCR and PCR-Southern blotting analysis. TrxS gene was expressed in transgenic plants by RT-PCR analysis. The activity of Trxh and α-amylase of transgenic line were higher than that of non-transgenic line, which is helpful to improve malting quality of barley.     

Effect of Transgenic Corn Cultivation and Sampling Location on Feeding Habits of Collembola

Transgenic corn was modified to combat lepidopteran pests, herbicide and other deteriorate environmental factors. However, fewer studies concerned the effects of transgenic corn cultivation on the soil Collembola in black soil region in China. In Jilin black soil region, a six treatments study including three corn varieties （transgenic corn, control and local variety） and two sampling locations （in rizhosphere and in between plants） were deployed to study the effect of transgenic corn cultivation and sampling location on densities, species richness and community structure of Collembola. We found that root biomass was marginally decreased in transgenic corn and control variety when compared to local variety. Collembola significantly affected by sampling location treatments. Densities and species richness of collembolans significantly decreased in ＂between plants＂ than in ＂rizhosphere＂. Densities of Collembola were impacted by corn varieties. In the whole growing season, densities of Collembola significantly decreased in control than in local variety. Collembolans in transgenic variety only slightly decreased in ＂jointing stage＂ than local variety. Collembolans adversely affected in ＂between plants＂ when compared to ＂in rhizosphere＂ in ＂seeding stage＂ but not other stages. These variations of collembolans were in lines with the trend of corn root biomass. It is indicated that Collembola variations could be caused by food resources （root biomass） but not corn varieties （transgenic, control and local）.     

Silencing of OsXDH reveals the role of purine metabolism in dark tolerance in rice seedlings

Xanthine dehydrogenase(XDH) is a crucial enzyme involved in purine metabolism. To evaluate the effect of XDH deficiency on rice growth during dark treatment, wild type(WT) Nipponbare(Oryza sativa L.) and two independent transgenic lines with severe RNAi suppression(xdh3 and xdh4) were used in the present experiment. Under normal growth conditions, chlorophyll levels and biomass were indistinguishable between WT and the two RNAi transgenic lines, but XDH enzyme activity and ureide levels were suppressed in XDH RNAi transgenic lines. When XDH RNAi transgenic lines were subjected to dark treatment, chlorophyll content and biomass were significantly decreased, while O_2~–· production rate and malonaldehyde(MDA) were significantly increased compared to WT. The spraying test of exogenous allantoin raised chlorophyll content and biomass and reduced O_2~–· production rate and MDA in WT and both transgenic lines, and it also simultaneously reduced differences between RNAi and WT plants caused by XDH deficiency in growth potential and anti-oxidative capacity under dark treatment. These results suggested that fully functional purine metabolism plays an important role in reducing the sensitivity of rice seedlings to dark stress.     

Purification of Aminopeptidase N Protein and Differences in cDNAs Encoding APN1 Between Susceptible and Resistant Helicoverpa armigera Strains to Bacillus thuringiensis Toxins

The brush border membrane vesicles (BBMVs) in midgut of Helicoverpa armigera were successfully separated, and most of the Aminopeptidase N (APN) activities in BBMV were preserved. The 3-[(3-chlor-amidopropyl) dimethylammonio]-l-propane-sulphonate (CHAPS) can enhance the dissolution of BBMV, and phosphatidylinositol-specific phosopholipase C (PI-PLC) can cleave the APN from midgut membrane. The APN was primarily purified using a Mono-Q column. The results of immunoblotting showed that the 120 and 170 kDa proteins in the BBMV could bind CrylAc, and 120 kDa APN was a glycosylphosphalidylinositol(GPI)-anchored protein. Two Bt-resistant strains (Bt-P, Bt-M) were obtained after being selected for more than five years in laboratory using Bt insecticides and Bt transgenic cotton incorporated into diet separately. The resistance of Bt-P and Bt-M were 1 083.3 and 48.7 times that of susceptible strain. The genes encoding APNI in midgut of susceptible and resistant H.armigera were cloned by PCR and RACE techniques. The inferred amino acid sequences of APNI possessed the common character of APN family in insects. In comparison with APNI in susceptible strain, three nucleotide mutations were observed in the APNI of Bt-M strain and resulted in two amino acid replace in the putative protein sequences, and eight nucleotide mutations were observed in Bt-P strain and resulted in five amino acid replace.     

Performance of autoregulatory Senescence-inhibition Gene in Rice

The Performance of autoregulatory senescence - inhibition gene PSAG12 - IPT in rice has been investigated in the study. 422 transgenic plants from 134 independent resistant calli were ohmined from 4 rice varieties through Agrobacterium -mediated transformation. Among them, 233 were positive PSAG12 - IPT transgenic plants identified by GUS histochemical assay and PCR analysis.Southern analysis shorted the transgene was randomly integrated into rice genome, of which 42.29% was single copy. Investigations on photmynthesis function and agronomic characters R1 generation shorted that chlorophy Ⅱ content and photmynthesis rate of flag leaves in transgenic plants, were 41.23% and 50.24% higher than the control wild-type rice, respectively. The growth duration and plant height of the transgenic plants were similar to the control. Variations of other characters were dependent on the varieties. For the variety Millin with significant aging phenomenon in China, its total grains per hill, its seed setting rate and 1000-grain wdght were increased by 40.44%, 8.05% and 8.32% respectively. The results indicated that after leaf senesomce of varieties liable to age was delayed, the seed setting rate and the filling degree of seeds were improved, which finally resulted in significantly increased seed yield and biomass per hill. The new variety Wuyujing 2 without serious aging problem, was also increased in the panicles per hill, the totslgrains per hill, the seed yidd per hill and biomass in different degrees.     

Correlation of production constraints with the yield gap of apple cropping systems in Luochuan County,China

Apple occupies a dominant position in fruit production globally, and has become the main income source of local smallholder farmers in Luochuan County in the Loess Plateau area, one of the largest apple production areas in China. However, the annual productivity of apple orchards in this region remains low and has gradually declined over the years. The distinction and correlation of production constraints can contribute to the promotion of apple orchard productivity and the development of a sustainable orchard system. In the present study, survey data from 71 smallholder farmers were analyzed using a yield gap model to distinguish the production constraints and determine their correlation with the yield gap based on the structural equation model(SEM). The results indicated that the average apple yield in Luochuan County was 29.9 t ha~(–1) yr~(–1), while the attainable yield(Y_(att); the highest yield obtained from the on-farm surveys) was 58.1 t ha~(–1) yr~(–1). The average explained and unexplainable yield gaps were 26.3 and 1.87 t ha~(–1) yr~(–1). According to the boundary line analysis, crop load,number of sprayings and base fertilizer N were the top three constraints on apple production in 9.8, 7.8 and 7.8% of the plots, respectively. Among the production constraints, crop load and fruit weight affected apple yield through direct pathways,whereas other constraints influenced apple yield through an indirect pathway based on the SEM, explaining 51% of the yield variance by all the main production constraints. These results can improve the current understanding of production constraints and contribute to the development of management strategies and policies for improving apple yield.     

Molecular Identification and Cultivar Fingerprints of Prunus persica （L.） Batsch Germplasms

[Objective] The aim was to study the molecular identification and cultivar fingerprints of Prunus persica （L.） Batsch germplasms.[Method] Sixty peach genotypes,representing China common local cultivars and European samples were screened by microsatellites （simple sequence repeats,SSRs） and Inter-Simple Sequence Repeat （ISSR） markers.[Result] 26 reproducible bands were amplified by Nine SSR primers,and 24 of which were polymorphic; 236 bands were amplified by 30 ISSR primers,and 113 of which were polymorphic.31 genotypes were discriminated with 1-3 distinct polymorphic bands generated from the primers ISSR and SSR.Seven cultivar-specific ISSR fragments and two SSR unique alleles obtained from this study were available to be converted into Sequence Characterized Amplified Region （SCAR） markers.The genetic similarity coefficient （GS） estimated from these molecular data averaged were 0.939 （ranged from 0.856 to 0.983） for ISSR and 0.646 （ranged from 0.240 to 1.000） for SSR,respectively.The combined grouping association indicated that most local Chinese peach cultivars and exotic accessions were clustered together.This could be related to the mode of introduction and maintenance of the peach cultivars involving limited foundation germplasm,exchange of cultivars between plantations,and periodic development of new recombinant cultivars following sexual reproduction.[Conclusion] The results obtained in this work would help to improve the conservation,molecular identification and management of peach germplasm in breeding.     

Genetic Parameters Evaluation of Main Reproductive Traits for the New Yorkshire Dam Line with High Prolificacy

The genetic parameters of main reproductive traits for the new Yorkshire line with high prolificacy were evaluated by multiple animal model DFREML The re- suits showed that the heritability evaluates of the total number of born （TNB）, alive number of born （ANB）, litter weigh at birth （LWB） and litter weight at age of 21 days were 0.10, 0.15, 0.09 and 0.14, respectively. The genetic correlations between these traits, for example, the correlations between TNB/ANB, NB/LWB, TNB/LW21, ANB/LWB, ANB/LW21 and LWB/LW21 were 0.87, 0.74, 0.43, 0.69, 0.59 and 0.51, respectively.     

Levels of Germinable Seed in Topsoil and Yak Dung on an Alpine Meadow on the North-East Qinghai-Tibetan Plateau

In order to clarify the interactive mechanism between grazing yak and alpine meadow on the Qinghai-Tibetan Plateau,our study assessed seed density（by species） in the topsoil of alpine meadow with different grazing intensities in the Tianzhu area,north-eastern margins of the Qinghai-Tibetan Plateau and their rates of occurrence in yak dung.Seed density in the topsoil of the lightly grazed,moderately grazed,heavily grazed and extremely grazed alpine meadows in November,2010 were 1 551,1 692,2 660 and 1 830 grains m-2,while in the same meadows in April,2011 densities were 1 530,2 404,2 530 and 2 692 grains m-2,respectively.In the cold season pasture,mean seed density in yak dung from November to April in the lightly grazed,moderately grazed,heavily grazed and extremely grazed sites were 121,127,187,and 120 grains kg-1of dry yak dung.The proportion of total seed numbers in yak dung to soil seed bank in lightly grazed,moderately grazed,heavily grazed and extremely grazed alpine meadow was 1.40,2.62,0.69,and 0.90%.12 species out of the 47 were not found in topsoil but were found in yak dung,10 species out of 45 were not found in yak dung but were found in the topsoil.Endozoochorous dispersal by yaks is therefore very important for soil seed bank and plant biodiversity and population dynamics in alpine meadows.     

Strategies and Suggestions for the Development of Mechanized Rapeseed Production in Sichuan

Mechanization is the future direction of rapeseed production in Sichuan. A series of technology tests for rapeseed mechanized production have been carried out in terms of variety selection, key agronomic measures and machinery in the main production area of Sichuan in recent years. In the research, the agronomic techniques, including variety selection for mechanical production, key agronomic measures and mechanical harvesting were analyzed. Based on the present condi- tions, strategy and suggestions for the development of mechanized rapeseed produc- tion in Sichuan also were presented.     

Yeast-Derived β-1,3-Glucan Substrate Significantly Increased the Diversity of Methanogens During In vitro Fermentation of Porcine Colonic Digesta

The β-1,3-glucan from yeast has been extensively examined for its immuno-enhancing effects in animals.However,investigation on the relationship among β-glucan,gut microbiota and immune-modulating effects remains limited particularly in pigs.Considering the critical roles of gut methanogens in the microbial fermentation,energy metabolism and disease resistance,we investigated the phylogenetic diversity of methanogens from fermented cultures of porcine colonic digesta with（G） or without（N） yeast β-glucan based on sequences of the archaeal 16S rRNA gene.A total of 145 sequences in the G library were assigned into 8 operational taxonomic units（OTUs） with the majority of sequences（114/145） related to strains Methanobrevibacter millerae or Methanobrevibacter gottschalkii with high identities ranging from 97.9 to 98.6%,followed by 23 sequences to Methanobrevibacter ruminantium,2 sequences to Methanobrevibacter smithii and one sequence to Methanobrevibacter wolinii.The 142 sequences in the N library were assigned to 2 OTUs with most sequences（127/142） related to strains M.millerae or M.gottschalkii with sequence identities ranging from 97.9 to 98.5%,and 15 sequences related to M.gottschalkii with 97.9% identity.Shannon diversity index showed that the G library exhibited significantly higher archaeal diversity（P0.05） and Libshuff analysis indicated the differences in the community structure between the two libraries were significant（P0.0001）.In conclusion,the current study provides evidence that addition of yeast β-glucan significantly increased the diversity of methanogens in in vitro fermented porcine colonic digesta.     

Induced in vitro Expression of Human Lactoferrin in Goat Mammary Gland Epithelial Cell

[Objective]The aim of this study was to explore the technical system of induced expression in vitro of goat mammary gland epithelial cell,and evaluate expression efficiency of mammary gland specific vector and foreign protein at the cell level.[Method]Goat mammary gland epithelial cell transfected by human lactoferrin gene was inducted by culturing in DMEM/F12 medium supplemented with 5 mg/L insulin,5 mg/L prolactin and 1 mg/L hydrocortisone.Supernatant was collected per 6 hours and concentrated.Expression situation of foreign protein were detected by SDS-PAGE and Western blotting.[Result]There was target protein expression in the induced culture medium,which molecular weight was about 42 kD.[Conclusion]The method used in this study can induce goat mammary gland epithelial cell to express foreign gene,it lays a foundation for researching heterologous expression of foreign gene and producing mammary gland bioreactor.     

Influences of Cold Storage at 5℃ for Different Durations on Oviposition of Female Silkworm Moths

In order to investigate the resistance of female silkworm moths to cold storage and their copulation and oviposition after cold storage, copulation and ovipo- sition experiments of Liangguang 2 silkworms and Guican 1 female silkworm moths were conduced under cold storage at 5 ~C for different durations. The results indi- cated that, after cold storage at 5 ~C with relative humidity of 75%, the resistance of female silkworm moths to cold storage varied with different varieties; to be specific, 9-Fu and 7.Xiang, parent silkworm varieties of Liangguang 2 with ancestry of mul- tivoltinism, were relatively resistant to cold storage, and their copulation capability was still at a high level after cold storage for 72 h. Oviposition capability also varied with different varieties and durations; to be specific, under 5 ~C conditions, between two parent silkworm varieties of Liangguang 2, female silkworm moths of 9-Fu achieved the best results within cold storage duration of 24 h, female silkworm moths of 7.Xiang achieved the best results within cold storage duration of 72 h; be- tween two parent silkworm varieties of Guican 1, female silkworm moths of Xinhang achieved the best results without cold storage, female silkworm moths of 826 achieved the best results within cold storage duration of 48 h.     

Statistical Analysis of Regularity of Pesticide Residues in Vegetables Produced in Inner Mongoli

[Objective] The paper was to study regularity of pesticide residues in veg- etables produced in Inner Mongolia. [Method] Mathematical statistic analysis was carried out on 6 800 samples of veggies, fruits, leguminous vegetables, nuts and seeds produced in Inner Mongolia. [Result] The dominated vegetables in Inner Mon- golia were green leafy vegetables, solanaceous vegetables and melon vegetables, and their yields accounted for 70% of the total vegetables production. Since 2003, with the rapid increase of detected vegetable numbers, the status of vegetable qual- ity safety had entered into a new stage of sustainable steady after a rapid decline or periodic fluctuation, and the differences in safety levels were obvious, of which the range of exceeding standard rate of pesticide residues in solanaceous vegeta- bles, melon vegetables and leguminous vegetables （about 55% of the total vegeta- bles） was under 2%, with average values of 1.1%, 1.6% and 3.1%, respectively. They belonged to stable type. The exceeding standard rate of pesticide residues in green leafy vegetables and Chinese cabbage group （about 30% of total vegetables） presented a decreasing trend year by year, wondering in the range of 7%-10%. They belonged to main risk type. The time period of exceeding standard of pesticide residues in root vegetables and cole vegetables was under predictable, and its ex- ceeding standard rate in some years was over 5% （amplitude variation is over 15 percentage points）. They belonged to random risk type. The kinds of pesticides, which exceeding standard rate in vegetables were relatively intensive, presented vari- ance in different vegetable species. 70% of the pesticides belonged to intermittent over-limits and the probability was below 5%. About 20% tradition pesticides often exceeded standard and their probabilities were over 30%. The exceeding standard of organophosphorus and carbamates pesticides in vegetables presented a decreasing tend, while the risk of some new pesticides containing nitrogen element and high effective pyrethroids showed an increasing tendency. [Conclusion] The paper provid- ed decision making services for agricultural management departments.     

Genetic Analysis and Molecular Mapping of an All-Stage Stripe Rust Resistance Gene in Triticum aestivum-Haynaldia villosa Translocation Line V3

Triticum aestivum-Hayaldia villosa translocation line V3 has shown effective all-stage resistance to the seven dominant pathotypes of Puccinia striiforms f.sp.tritici prevalent in China.To elucidate the genetic basis of the resistance,the segregating populations were developed from the cross between V3 and susceptible genotype Mingxian 169,seedlings of the parents and F 2 progeny were tested with six prevalent pathotypes,including CYR29,CYR31,CYR32-6,CYR33,Sun11-4,and Sun11-11,F 1 plants and F 3 lines were also inoculated with Sun11-11 to confirm the result further.The genetic studied results showed that the resistance of V3 against CYR29 was conferred by two dominant genes,independently,one dominant gene and one recessive gene conferring independently or a single dominant gene to confer resistance to CYR31,two complementary dominant genes conferring resistance to both CYR32-6 and Sun11-4,two independently dominant genes or three dominant genes（two of the genes show cumulative effect） conferring resistance to CYR33,a single dominant gene for resistance to Sun11-11.Resistance gene analog polymorphism（RGAP） and simple-sequence repeat（SSR） techniques were used to identify molecular markers linked to the single dominant gene（temporarily designated as YrV3） for resistance to Sun11-11.A linkage map of 2 RGAP and 7 SSR markers was constructed for the dominant gene using data from 221 F 2 plants and their derived F 2：3 lines tested with Sun11-11 in the greenhouse.Amplification of the complete set of nulli-tetrasomic lines of Chinese Spring with a RGAP marker RG1 mapped the gene on the chromosome 1B,and then the linked 7 SSR markers located this gene on the long arm of chromosome 1B.The linkage map spanned a genetic distance of 25.0 cM,the SSR markers Xgwm124 and Xcfa2147 closely linked to YrV3 with genetic distances of 3.0 and 3.8 cM,respectively.Based on the linkage map,it concluded that the resistance gene YrV3 was located on chromosome arm 1BL.Given chromosomal location,the reaction patterns and pedigree analysis,YrV3 should be a novel gene for resistance to stripe rust in wheat.These closely linked markers should be useful in stacking genes from different sources for wheat breeding and diversification of resistance genes against stripe rust.     

Effects of Grafting on Root Exudates of Cucumber and Rhizosphere Environment under Copper Stress

[Objective] This study to aimed to investigate the effects of Cu stress on root exudates and microbial activities in rhizosphere of grafted and ungrafted cucum-ber seedlings, and therefore to elucidate the microbial mechanism of grafting for in-creasing cucumber plants tolerance to Cu stress [Method] Four treatments： （1） un-grafted seedlings ＋ test soil （U0）; （2） ungrafted seedlings ＋ test soil ＋ CuSO4·5H2O （U1）; （3） grafted seedlings ＋ test soil （G0）; （4） grafted seedlings ＋ test soil ＋ Cu-SO4·5H2O （G1） were set in the pot culture experiment. The contents of free amino acids, organic acids, phenolic acid and sugars, microbial population and enzyme ac-tivity in the four treatment were measured, respectively. [Result] The secretion of amino acids and organic acids were increased under Cu stress. The amino acids secretions of grafted seedlings roots were obviously higher than ungrafted seedlings except for Phe and Val. At the same time, the secretion of oxalic acid, malic acid, acetic acid, citric acid, cinnamic acid, ρ-hydroxybenzoic acid and benzoic acid of grafted seedlings were significantly higher than ungrafted seedlings as wel . There-fore, more Cu2＋ were restricted in soil by chelating, complexing and precipitation with root exudates, and its toxicity was decreased. The soil microbial biomass C and N in grafted cucumber rhizosphere were significantly higher than those in ungrafted cu-cumber rhizosphere, whereas basal respiration and metabolic quotient were signifi-cantly lower. Under Cu stress, the numbers of actinomyces and nitrogen fixing bac-teria decreased and the number of fungi increased significantly, whereas there was no significant difference in amounts of bacteria. The numbers of bacteria, actino-myces, and nitrogen fixing bacteria in grafted cucumber rhizosphere were significant-ly higher than those in ungrafted cucumber rhizosphere, but the number of fungi was opposite. The activities of soil urease, phosphatase, sucrase and catalase in grafted cucumber rhizosphere were significantly higher than those in ungrafted cu-cumber rhizosphere. [Conclusion] These indicated that the soil microbial environment and soil enzymes activities were improved by grafting under Cu stress, and as a re-sult, the adaptability of cucumber to Cu stress was improved.