Scientia Agricultura Sinica

Advances in Study on Food Legumes Resistance Breeding in China

DUAN Can-Xing, ZHU Zhen-Dong, SUN Su-Li, WANG Xiao-Ming

Scientia Agricultura Sinica. 2013, 46(22): 4633-4645. doi:10.3864/j.issn.0578-1752.2013.22.001

Abstract ( 438 )

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As a kind of traditional food resources of human, food legumes are important crops in adjustment of planting structure, foreign exchange-earning and increasing the income of mountain farmers in China. Diseases and insect pests often lead to a serious decline in yield and quality of food legumes. The cultivation and utilization of resistant cultivars is the most economical, safe and effective method for controlling pests. So for, more than ten thousand food legumes germplasm have been identified and evaluated for resistance to diseases and pests in China, and some resistant accessions were screened out. Resistance inheritance and gene mapping were conducted. Some resistant varieties were cultivated and deployed. However, compared with the major crops, weak basal and lagged researches on the food legumes resistance breeding result in lack of cultivars with high-multi resistance and high yield and quality. In this paper, studies on food legume germplasm screening and evaluation for resistance, resistance inheritance, discovery and mapping of resistance genes and breeding resistant cultivar were reviewed. Several important directions for the future researches were prospected. The main aim is to supply useful information so as to promote the development of breeding for resistant food legume cultivars in China.

Identification of Soybean Maturity SSSLs and QTL Mapping Based on Meta-Analysis

CHE Jing-Yu-12, LIU Chun-Yan-3, JIANG Hong-Wei-13, HAN Xue-3, MAO Yan-Zhi-12, XIN Da-Wei-1, CHEN Qing-Shan-1, HU Guo-Hua-3

Scientia Agricultura Sinica. 2013, 46(22): 4646-4656. doi:10.3864/j.issn.0578-1752.2013.22.002

Abstract ( 388 )

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【Objective】Soybean maturity is a quantitative trait controlled by multiple genes, which is a key trait affecting soybean yield and adaptability. The heredity of soybean maturity with single segment substitution lines (SSSLs) was studied and the main effect QTLs of soybean maturity were identified. 【Method】 Fifteen sets of backcross introgression lines were constructed by using Hongfeng 11 as recurrent parent and 15 soybean varieties as the donor parent, respectively. Based on the results of meta-analysis of soybean maturity (R8), the SSR markers from real QTL interval were used to identify SSSLs. Using the method of figure genotype calculated the length of substitution fragment, using the statistical software SPSS analyzed the main effect QTLs of soybean maturity. 【Result】Sixteen substitution fragments were detected on linkage group C2 and L. Among them, 7 substitution fragments were detected on linkage group C2, substitution fragment length was 9.8cM in total. Nine substitution fragments were detected on linkage group L, and the substitution fragment length was 37.212 cM in total. Previous researches show that Sat_010 and Satt156 are the specific markers for E4/e4 on linkage group L, and 5 QTLs（Sat_238, Satt460, Sct_010, Satt166, and Sat_113 ） were identified on the maturity in one-way. Single fragment Satt460 could shorten the growth period of soybean, and single fragments Sat_238, Sct_010 and Sat_113 could delay soybean growth stages. Sat_238 Satt460, Sct_010, Satt166, and Sat_113 were identified to be the important SSR markers on soybean maturity. 【Conclusion】Five markers from meta-analysis were identified as the important markers on soybean maturity. The single fragment Satt460 could shorten the soybean growth period, single segments Sat_238, Sct_010, and Sat_113 could delay soybean growth period.

Molecular Mechanism Underlying Photoperiodic-Induced Potato Tuber Formation

XIE Ting-Ting, LIU Jun

Scientia Agricultura Sinica. 2013, 46(22): 4657-4664. doi:10.3864/j.issn.0578-1752.2013.22.003

Abstract ( 508 )

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Research on potato tuber formation is not only an important aspect of plant developmental biology, but also critical to improve potato yield and quality. It has been known for a long time that photoperiod plays a vital role in potato tuberization. Molecular mechanism underlying it, however, had made less progress until this century. More and more data support that potato tuber formation and Arabidopsis flowering share a similar pathway and a large number of common genes, such as Phytochrome, CONSTANS (CO), FLOWERING LOCUS T (FT), LOV blue light receptors and transcription factor CDF. In addition, a homeobox gene POTH1 and its interacting gene StBEL5, which were first studied in potato tuber formation, are identified to be affected by light. In this paper, recent progress in molecular mechanism underlying photoperiodic-induced potato tuber formation is described in the aspects of light signal perception, photoperiodic signal transduction and tuber induction by mobile signal molecules.

Effect of Post-Anthesis Alternate Wetting and Drying Irrigation on Protein Expressions in Superior and Inferior Spikelets of Rice

CHEN Ting-Ting, CHU Guang, HUA Xiao-Long, PAN Yan, WANG Zhi-Qin, YANG Jian-Chang

Scientia Agricultura Sinica. 2013, 46(22): 4665-4678. doi:10.3864/j.issn.0578-1752.2013.22.004

Abstract ( 428 )

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【Objective】This study aimed to understand the mechanism underlying the effect of alternate wetting and drying irrigation on grain filling of rice. 【Method】Two rice cultivars, Liangyoupeijiu (two-line indica hybrids) and Yangjing 4038 (japonica), were grown in cement tanks. Three irrigation regimes, conventional irrigation (CI), alternate wetting and moderate drying irrigation (WMD) and alternate wetting and severe drying irrigation (WSD), were imposed from full heading to maturity. The expressions of the proteins in superior and inferior spikelets were investigated using the method of two-dimensional gel electrophoresis.【Result】No significant differences in grain-filling rate and grain weight were observed for the earlier-flowering superior spikelets among the three irrigation regimes. For the later-flowering inferior spikelets, however, their grain-filling rate and grain weight were significantly increased under the WMD, but significantly reduced under the WSD when compared with those under the CI. Both WMD and WSD showed no significant impact on the protein expressions in superior spikelets. However, the WMD enhanced expressions of many proteins, such as orthophosphate dikinase, glyceraldehyde-3-phosphatedehydrogenase, 5-methyltetrahydropteroyltriglutamate-homocysteine methyltransferase, S-adenosylmethionine synthetase, glyoxalase I and manganese superoxide dismutase in inferior spikelets, whereas the WSD inhibited expressions of these proteins. The WSD also up-regulated expressions of some proteins, such as the proteins related to the inhibition of signal transduction and energy metabolism. The two cultivars exhibited the same tendencies. 【Conclusion】 Expression differences of the proteins related to grain filling in inferior spikelets account for their increases or decreases in grain-filling rate and grain weight under the WMD or WSD.

Morphology and Physiological Characteristics of Cultivars with Different Levels of Cold-Resistance in Winter Rapeseed (Brassica campestris L.) During Cold Acclimation

LIU Zi-Gang, SUN Wan-Cang, YANG Ning-Ning, WANG Yue, HE Li, ZHAO Cai-Xia, SHI Peng-Fei, YANG Gang, LI Xue-Cai, WU Jun-Yan, FANG Yan, ZENG Xiu-Cun

Scientia Agricultura Sinica. 2013, 46(22): 4679-4687. doi:10.3864/j.issn.0578-1752.2013.22.005

Abstract ( 550 )

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【Objective】The objective of this study was to provide an evaluation method for the development of cold tolerant Brassica campestris germplasm. 【Method】 In this study, the plant morphological characteristics and physiological parameters were measured, and the molecular mechanism of cold-resistance of several cultivars was discussed. 【Result】 Under low temperature, the majority of the stomata in cold sensitive cultivars were closed or semi-closed. But the stomata in strong cold-tolerant cultivars (Longyou 6) maintained opening. The photosynthetic rate and root/shoot ratio were higher, dry matter accumulation in root of Longyou 6 was higher than those of other cultivars. Simple correlation analysis indicated that the soluble protein content was positively correlated with cold-tolerance, but no significant correlation was observed between cold-tolerance and SOD activity, POD activity, CAT activity, free proline content or MDA content. At low temperature, in comparison with cold tolerant variety Tianyou 2, the ice crystals in apoplast proteins extracting solution of ultra strong cold resistant variety Longyou 7 were slight and symmetrical. The shape of ice crystal was pentagon, elliptocytosis or bipyramidal. 【Conclusion】 During the pre-winter cold acclimation stage, the soluble protein content in leaves of strong cold-tolerant winter rapeseeds was high, and the majority of stomata in leaves remained opening, the photosynthetic rate was high, too. Photosynthates might be preferentially transported and stored in the roots. The expression of the antifreeze proteins (AFPs) was up-regulated in roots, which might ease the damage to cells in roots caused by restraining extracellular frozen.

The Relationship Between the Contents of Nutrients and Tannins in Different Cotton Varieties and Their Resistance to Apolygus lucorum

YANG Yu-Hui, ZHANG Qing-Wen, LIU Xiao-Xia

Scientia Agricultura Sinica. 2013, 46(22): 4688-4697. doi:10.3864/j.issn.0578-1752.2013.22.006

Abstract ( 438 )

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【Objective】 The objective of this study is to clarify the relationship between the contents of protein, soluble sugar and tannin in different tissues along developmental stage of various cotton varieties and their resistance to Apolygus lucorum.【Method】 By using the 12 cotton varieties as materials, with the method of resistance appraising in the field for two consecutive years and with the Bradford method, Anthrone-sulfuric acid method and Vanillin method, tests were made in laboratory to make clear the relationship between the contents of protein, soluble sugar and tannin in different tissues along developmental stage of various cotton varieties and their resistance to A. lucorum.【Result】The resistant level of different cotton varieties was obviously different, Yunnanhuangrongtumian and Meizhongmian were high resistant varieties (integrate resistance index＜1), AH-510, Sulianmian, Jing 7516-1 and Chizhou red leaf were low resistant varieties (integrate resistance index＞3). The content of protein in bud in budding period and boll of bolling period decreased with the increase of resistance of the cotton varieties to A. lucorum. The content of soluble sugar in leaf in seeding period increased with the increase of resistance of the cotton varieties to A. lucorum, but in bud in budding period decreased with the increase of resistance of the cotton varieties to A. lucorum. The content of tannin in bud in budding period decreased with the increase of resistance of the cotton varieties to A. lucorum, but in boll in bolling period increased with the increase of resistance of the varieties to A. lucorum. 【Conclusion】Yunnanhuangrongtumian had the highest resistance levels to A. lucorum, and Chizhou red leaf had the lowest resistance levels to A. lucorum. The content of protein in bud in budding period and boll in bolling period indicated a clear negative linear correlation with the resistance to A. lucorum. The content of soluble sugar in leaf in seeding period indicated a clear positive linear correlation with the resistance to A. lucorum, but in bud in budding period indicated a clear negative linear correlation with the resistance to A. lucorum. The content of tannin in bud in budding period indicated a clear negative linear correlation with the resistance to A. lucorum, but in boll in bolling period indicated a clear positive linear correlation with the resistance to A. lucorum.

Effects of Biofumigation with Rapeseed Meal on Disease Control of Phytophthora Blight of Chilli Pepper

MA Yan, HU An-Yi, YANG Hao, CHANG Zhi-Zhou, XU Yue-Ding, ZHANG Jian-Ying

Scientia Agricultura Sinica. 2013, 46(22): 4698-4706. doi:10.3864/j.issn.0578-1752.2013.22.007

Abstract ( 478 )

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【Objective】The objective of this study is to research the effects of hydrolysis product of rapeseed meal (RSM) on growth inhibition of Phytophthora capsici and on disease control of Phytophthora blight of chilli pepper cultivation in greenhouse. 【Method】 Inhibition effect of hydrolysis product of RSM on mycelium and zoospore was studied by in vitro test in dishes and imitated biofumigation in container, and disease control effects of biofumigation with RSM2 on Phytophthora blight of chilli pepper in pot and field were also carried out. 【Result】 Volatiles and non-volatiles released from RSM hydrolysis inhibited mycelium growth and zoospore germination in various degrees, and the inhibition effect of RSM2 was better than that of RSM1. The inhibition effect of volatiles produced from RSM hydrolysis on zoospore was stronger than on mycelium of P. capsici while non-volatiles displayed stronger inhibition effect on mycelium than on zoospore. The inhibition effect of volatiles and non-volatiles showed no significant difference on mycelium but on zoospore of P. capsici. Soil fumigation in sealed container was carried out in laboratory at different application rates of RSM and the results showed that volatiles released from RSM hydrolysis killed mycelium of P. capsici when RSM was added to soil at the rate of 0.2% (w/w). In a pot experiment, real-time PCR was adopted to assess the population of P. capsici. The results showed that the efficacy of disease control of RSM treatment for Phytophthora blight was 100%, the same as dazomet treatment, although P. capsici exsisted in soil at density of 36 zoospore/g soil. RSM2 treatment not only increased the population of fungi and actinomycetes but also increased the total number and diversity of soil microbe. The plot experiment conducted in greenhouse showed that the average value of control efficacy for Phytophthora blight was 82% and the yield of chilli pepper increased with 16.4% in two continuous cropping seasons. The effect of RSM treatment in chilli pepper production in protected field was better than dazomet treatment. 【Conclusion】RSM2 showed great inhibition effect on growth of P. capsici. The treatment of biofumigation with RSM2 suppressed disease incidence of Phytophthora blight in field efficiently and increased yield of chilli pepper.

Application of Pseudo-Ternary Phase Diagram for Screening the Formula of 25% Fluopicolide•Pyraclostrobin EW

ZHANG Peng-12, HUANG Qi-Liang-1, WANG Wen-Qiao-2, CAO Li-Dong-1, LI Feng-Min-1

Scientia Agricultura Sinica. 2013, 46(22): 4707-4715. doi:10.3864/j.issn.0578-1752.2013.22.008

Abstract ( 455 )

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【Objective】The objective of this study is to explore the application of pseudo-ternary phase diagram for screening the formula of emulsion in water (EW). 【Method】 Blank and drug-containing pseudo-ternary phase diagrams were drew and corresponding samples were prepared and their quality indexes were determined on the basis of the mixing ratio of fluopicolide and pyraclostrobin was confirmed. The effect of the types of emulsifiers and technical materials on the size of the emulsion area in phase diagram and the effect of the size of the emulsion area in phase diagram on the stability of the EW samples were explored. The range of dosage of emulsifiers to processing EW were defined by the way of observing the change of phase states and determining the quality indexes of the samples which were processed by screening different points from the pseudo-ternary phase diagram.【Result】The results demonstrated that though the emulsifiers of EW couldn’t be screened only according to the area of the emulsion area in pseudo-ternary phase diagram, the emulsifiers which formed bigger emulsion area in pseudo-ternary phase diagram were more suitable for EW processing. The content of SAA phase in EW had relations with the content of oil phase in practical formula and usually the ratios of SAA phase and oil phase ranged from 0.11 to 0.25 were more likely to processing out of stable EW. Suitable emulsifier dosage was helpful to reduce particle size of emulsion and improve stability of EW samples. In alliance with Xanthan gum, the use of organic bentonite could enhance the physical stability of the fluopicolide•pyraclostrobin EW. 【Conclusion】The method of pseudo-ternary phase diagram can define the types and dosages of emulsifiers which is suitable for EW processing quickly and conveniently and has guiding function to the formula screening of EW.

Comparison of Spatial Prediction of Soil Properties Under Different Sampling Sizes

CHAI Xu-Rong-1, HUANG Yuan-Fang-2

Scientia Agricultura Sinica. 2013, 46(22): 4716-4725. doi:10.3864/j.issn.0578-1752.2013.22.009

Abstract ( 432 )

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【Objective】The objective of this study was to investigate the impact of the sample size on the accuracy of the spatial prediction of soil properties.【Method】Soil organic matter(SOM), mass water content of soil, soil available potassium and available manganese were used as research objects, the methods of moments (MoM) and the residual maximum likelihood (REML) were used to compute the variogram of soil properties. Predictions based on the MoM and REML variograms were compared under different sampling sizes.【Result】Whether the MoM or REML was used, there was a significantly improved prediction accuracy for each soil variables with the increase of the sampling size from 50 to 70. As the number of samples gradually increased from 70 to 150, the prediction accuracy was not significantly improved. Predictions based on REML variograms were not more accurate than those from MoM variograms with the inreasion of sampling size from 50 to 150.【Conclusion】The number of samples has a significant impact on the accuracy of prediction of soil properties. When the number of samples is less than 70, the predictions are not reliable regardless of which variogram is used.

Contribution of Nitrification and Denitrification to the Nitrous Oxide Emission from Forest and Grassland Soils

LI Ping-1, LANG Man-2

Scientia Agricultura Sinica. 2013, 46(22): 4726-4732. doi:10.3864/j.issn.0578-1752.2013.22.010

Abstract ( 502 )

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【Objective】It is of great significance to explore the contribution of nitrification and denitrification to the N2O emission from forest and grassland soils under aerobic condition, and to compare the response of the contribution to temperature change.【Method】An laboratory aerobic (60% water holding capacity) incubation experiment was conducted at 10℃ and 15℃ to measure the atom 15N excess of ammonium, nitrate, and nitrous oxide using 15N isotope tracers, the contribution of nitrification and denitrification to the N2O emission was calculated. 【Result】 Under aerobic condition, both processes occurred simultaneously and were involved in N2O production, 53.1%–72.0% of N-N2O came from nitrification, which is the main source of N2O from soils. The average contribution of denitrification to N2O emission was 44.9% for forest soil, which is significantly higher than grassland soil (28.9%), while nitrification of forest soil contributed 55.1% to N2O emission, which is significantly lower than grassland soil (71.1%). Increasing temperature from 10℃ to 15℃ increased N2O emission significantly for both two soils, but had no effects on the relative contribution of nitrification and denitrification to N2O emissions.【Conclusion】 Microbial transformation of nitrification is the main source of N2O from soils under aerobic condition, however, the proportion of N2O emitted from denitrification should not be neglected.

Dispersal Limitation Versus Environment Filtering in the Assembly of Plant Communities in the Ziwu Mountains

WANG Shi-Xiong, GUO Hua, WANG Xiao-An, FAN Wei-Yi

Scientia Agricultura Sinica. 2013, 46(22): 4733-4744. doi:10.3864/j.issn.0578-1752.2013.22.011

Abstract ( 445 )

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【Objective】Improved regional eco-environment is critical to forming ecologically sustainable land-use policies and accelerating agricultural production. Restoring natural vegetation is one of the effective approaches to improve the ecological conditions, and understanding what governs community assembly is the first step, but has been a continuing challenge for ecologists.【Method】In the Ziwu Mountains of the Loess Plateau, how the community assembly is affected by environmental filtering and dispersal limitation was examined by using canonical variation partitioning and Mantel tests, i.e. partition the variation of community composition/dissimilarity distance between environmental and spatial factors/distance. Three species groups (herb layer, shrub layer and tree layer) that differ in traits of likely importance for environmental filtering and dispersal limitation (height and growth form) were analyzed. 【Result】 All forest types were simultaneously governed by environmental control and spatial processes; together, these processes explain 29% to 65% of the species composition variation (P<0.01). However, the effects of these two processes were varied among species groups; shrub layer and tree layer species were dominated by spatial process while there was similar importance between two processes for herb layer. Mantel and partial Mantel tests showed significant correlations between community dissimilarity and environmental dissimilarity/geographical distance (P<0.05). Herb layer showed a high correlation with environmental dissimilarity while shrub and tree layer displayed high correlations with geographical distance. 【Conclusion】 The assembly of plant communities in the Loess Plateau appears to be simultaneously driven by environmental filtering and dispersal limitation. This case study shows the importance of the dispersal limitation in understanding the forests assembly in the Loess Plateau. Consequently, it is important that management planning for restorations of natural vegetations take into account both habitat heterogeneity and geographical differences.

Control of Exogenous H2O2 on Dry Rot of Potato Tuber and Possible Mechanism of Action

HU Lin-Gang, LI Jian-Peng, LI Yong-Cai, BI Yang, GE Yong-Hong, WANG Yi

Scientia Agricultura Sinica. 2013, 46(22): 4745-4752. doi:10.3864/j.issn.0578-1752.2013.22.012

Abstract ( 509 )

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【Objective】The inhibitory effect of exogenous H2O2 against dry rot of potato and its mechanism of action were studied in this paper. 【Method】Mycelium growth, colonial morphology and hyphae ultrastructure of Fusarium sulphureum after treated with H2O2 were observed by scanning electron microscopy (SEM) and transmission electron microscopy (TEM) in vitro test, and the curative and preventative effect of exogenous H2O2 on dry rot of potato tuber (Longshu No.3) inoculated with F. sulphureum pre- and post-etreatment were also evaluated.【Result】Spore germination and mycelium growth of F. sulphureum were strongly inhibited by exogenous H2O2 in a concentration-dependent manner. Besides, exogenous H2O2 could dramatically increase the membrane permability of F. sulphureum. Morphological changes such as asymmetrical, distorted, broken and wizened hyphae were observed by scanning electron microscopy (SEM) observation. Transmission electron microscopy (TEM) observation further documented these changes, irregularly thickened hypha cell walls and cavity could be observed. Results of in vivo test showed that exogenous H2O2 was effective in the control of dry rot of potato tuber inoculated with F. sulphureum pre- and post-etreatment.【Conclusion】 H2O2 can directly suppress the pathogen, and effectively increase the disease resistance of potato tuber tissue, thus it represent a potential fungicides for the integrated control of postharvest diseases in potato tuber.

A Study of the Flavonoids in Different Tissues of the Basic Citrus Types Native to China

LI Li-Gai-1, XI Wan-Peng-13, ZHANG Yuan-Mei-1, JIAO Bi-Ning-2, ZHOU Zhi-Qin-13

Scientia Agricultura Sinica. 2013, 46(22): 4753-4762. doi:10.3864/j.issn.0578-1752.2013.22.013

Abstract ( 544 )

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【Objective】The aim of this study was to evaluate the flavonoid composition and content in flower, peel and leaf of the basic genotypes of genus citrus in an attempt to provide information for the utilization of resource.【Method】Eleven flavonoids in different organs (flower, peel, leaf) of 10 typical genotypes representing five basic types of the genus citrus were determined by UPLC-PDA.【Result】Naringin is the major flavonoid in Papeda and Cephalocitrus, and hesperidin is the major flavonoid in Citrophorum, Aurantium and Sinocitrus．The highest naringin content was detected in the flowers of Guangxishatianyou (11 987.1 mg•kg-1FW), whlie the highest hesperidin content was detected in the peel of Edanhongju (14 794. mg•kg-1FW). Significant differences were observed in falvoniod contents of different Citrus plant organs. The highest total contents of 11 flavonoids of flower, peel and leaf were Guangxishatianyou (12 198.5 mg•kg-1FW), Edanhongju (16 744.1 mg•kg-1FW）and Taiwanpenggan (13 396.3 mg•kg-1FW）, respectively. The flavonoids content of flowers, peel, leaves in different genus citrus showed different variation patterns. However, there was only an abundance difference among different organizations of the same genus citrus, the orders of total flavonoids content variation in Papeda, Citrophorum, Aurantium, Cephalocitrus, Sinocitrus were flower＞leaf ＞peel, flower＞peel＞leaf, peel＞flower＞leaf，flower＞leaf＞peel, peel＞leaf＞flower, respectively.【Conclusion】The citrus genotypes native to china are rich in flavonoids, and obvious variations were observed between different types and tissues. The utilization of citrus resources should be on the basis of their respective characteristics.

A Review of the Antioxidant Peptides Derived from Animal Protein

HUANG Ming, WANG Lu-Sha

Scientia Agricultura Sinica. 2013, 46(22): 4763-4773. doi:10.3864/j.issn.0578-1752.2013.22.014

Abstract ( 492 )

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Oxidation both influences the quality of food and human being’s health, so antioxidation has been the focus of life sciences for years. Nowadays, the antioxidant peptides derived from animal protein hydrolysates have drawn more and more attention. Because these peptides are being considered as potential sources to effectually control various oxidative processes(radical scavenging, inhibition of lipid peroxidation and metal ion chelation) in food as well as in the living body, and these peptides also can promote consumer health when they are used as functional ingredients. This review summarized the literature on the antioxidant peptides derived from animal protein hydrolysates. Furthermore, the factors which influence the antioxidative capacity of the antioxidant peptides, methods used for assessing antioxidant capacity and examples of prospective applications and challenges were also discussed.

Peptide Linkers Optimized Recombinant Enzyme Gene of XynB-ManA and Its Co-expression in pK15 Cells

ZHANG Xian-Wei, ZHANG Guan-Guan, WU Zhen-Fang, MENG Fan-Ming, LIU De-Wu, ZHANG Mao, XU Wei-Hua, ZHENG En-Qin, HE Xiao-Yan, LI Zhen, LI Zi-Cong

Scientia Agricultura Sinica. 2013, 46(22): 4774-4783. doi:10.3864/j.issn.0578-1752.2013.22.015

Abstract ( 421 )

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【Objective】This study was aimed at constructing a fusion enzyme of xylanase and mannase which functioned well in mammalian expression system for the research of transgenic animals and potential utilization in the industry of feed enzyme.【Method】Eleven chimeric genes of xylanase and mannase differed in linkers were constructed by SOE-PCR, and then subcloned into pcDNA3.1 vector. The cell free culture supernatants were analyzed by the dinitrosalicylic (DNS) acid method after the recombinant plasmids were transiently transformed into PK15 cells .【Result】Compared with the parental enzymes，the fusion enzymes with Linkers a3, pS3, and “self-cleavage”T2A displayed significantly higher catalytic efficiency. Specifically, the XynB-a3-ManA was increased by 54.06% for the xylanase and of 104.40% for the mannase in enzyme activity, and worked well at pH3.0-7.0 and was partially resistant in pH3.0-8.0.【Conclusion】The results demonstrated that the firstly obtained enhanced bifunctional fusion XynB-ManA enzyme which could be expressed and secreted in mammalian expression system by optimizing the peptide Linkers between two parental genes.

Characterization and Expression Analysis of Goat Ovarian chi-miR-100f-5p

NA Ri-Su-12, LIU Guang-1, XIANG Yang-1, ZHAO Hai-Lin-1, WU Ting-1, ZHAO Jin-Hong-2, LI Fan-1, HUANG Xuan-Yang-1, SUN Xiao-Wei-1, ZHENG Jiao-1, ZHANG Jia-Hua-1

Scientia Agricultura Sinica. 2013, 46(22): 4784-4790. doi:10.3864/j.issn.0578-1752.2013.22.016

Abstract ( 447 )

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【Objective】 The relationship between the goat ovarian expressed miR-100 with its predicted target genes and goat ovarian function or follicular development was studied in order to provide data for goat breeding mechanism.【Method】 Goat ovarian expressed miR-100 was obtained by cloning and sequencing a small RNA library with pooled RNA from goat ovarian tissue. The expression profiling was detected by RT-qPCR. miR-100 target genes and functional classification were predicted by bioinformatics method.【Result】The chi-miR-100f-5p, at a length of 21 nt , was isolated from goat ovaries. The sequence is highly homologous with dan-miR-100, and has one loci in the genome negative (-) chain, which located in goats sequence CM000899.1: 33997894 to 33997914; The chi-miR-100f-5p was detected in goat ovarian, pituitary, heart and other tissues, showing chi-miR- 100f-5p was not an ovarian-specific miRNA. It was found that chi-miR-100f-5p expressed at higher level in heart and pituitary, secondly in ovary, and the expression of chi-miR-100f-5p in the liver, spleen, lung, and kidney was significantly lower than in ovary (P＜0.01). It was alsofound that chi-miR-100f-5p expressed at significantly higher level in single lambing Inner Mongolia cashmere goat ovary than the high fecundity Dazu black goat ovary (P＜0.01); The chi-miR-100f-5p was mainly targeted for integrin β1 subunit and pituitary-specific transcription factor 1 gene, and might be involved in the integrin signaling pathway, promoting the growth and differentiation of pituitary endocrine cells, and regulating the hypothalamic-pituitary-gonadal axis hormonessecretion.【Conclusion】The newly discovered chi-miR-100f-5p in goat ovarian may be an important candidate miRNA in regulation of breeding.

Polymorphism of the 60149273th Loci on X Chromosome Among Fat Tail and Thin Tail Breeds and Its Gene Mapping

GAN Shang-Quan, SHEN Min, LI Huan, LIANG Yao-Wei, YANG Jing-Quan, GAO Lei, LIU Shou-Ren, WANG Xin-Hua

Scientia Agricultura Sinica. 2013, 46(22): 4791-4799. doi:10.3864/j.issn.0578-1752.2013.22.017

Abstract ( 444 )

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【Objective】 The objective of this paper is to study the polymorphism of the SNP located at the position of 60149247 on X chromosome between fat tail (rump) and thin tail breeds, and to provide the basic data for analysis of the mechanism of tail fat deposition in sheep.【Method】 Firstly, PCR-SSCP genotyping method was used to analyze genotypes among different tail types of sheep breeds, and definite allelic genes were cleared by PCR-direct sequencing, and the chi-square test was employed to detect the balance of genotype distribution in different breeds. Secondly, the SNP was mapped on sheep androgen receptor (AR) and the whole CDS sequences of AR were obtained by comparative genome and silicon cloning methods. Taking cDNA of Altay tail fat as DNA template, RT-PCR method was used to clone sheep AR spinning six exons, and sequence homology and phylogenetic tree were analyzed by bioinformatics methods. 【Result】The results of PCR-SSCP showed that there is no polymorphism in fat-tail sheep breeds (Hu sheep and Altay sheep), and the genotypes of the two breeds are all GG genotype. And there are three genotypes (GG, GA and AA) in thin-tail sheep populations (Chinese Merino sheep and Suffolk sheep), and the frequency of GG genotype dropped to 10% and 21%, respectively. The whole CDS sequences of sheep AR were cloned by silicon cloning, and sheep AR containing the fragment from the 3rd exon to 8th exon of AR was successfully cloned in total RNA of Altay tail fat for the first time, and the sequences are completely the same to the sequences predicted by silicon cloning. The sequence alignment also showed that AR is highly conserved in mammalian animals, and the homology was up to 90% among different mammalian animals, and the phylogenetic tree analysis showed that sheep is relatively close to bovine, pigs and dolphin. 【Conclusion】This study was the first to identify a novel SNP in the 3rd intron of the sheep AR associated with the trait of fat tail, and it hinted that the SNP will be used as a practical application of molecular markers in low-fat sheep breeding. At the same time, AR will also serve as an important candidate gene in studying tail fat deposition in Sheep.

Effects of Immune Stress on the Bacterial Community in the Broiler Chickens Gastrointestinal Tract Analyzed by Real-Time PCR

FENG Yan-12, YANG Xiao-Jun-2, LI Wu-Lin-2, YANG Ning-1, YAO Jun-Hu-2

Scientia Agricultura Sinica. 2013, 46(22): 4800-4807. doi:10.3864/j.issn.0578-1752.2013.22.018

Abstract ( 496 )

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【Objective】The purpose of this experiment was to investigate the effects of immune stress on intestinal microflora and common bacteria changes of broiler chickens. 【Method】 One hundred and eighty 1-day old AA broilers were allocated into 4 groups. Each treatment had 5 replicate cages of 9 broilers per replicate pen. Different immune stress models, including the control group (no vaccines, NV), conventional vaccines group (CV), immune hyperthyroidism group (LPS) and immune suppression group (CYP) were set in this study. On d 21, 28, 35 and 42, each treatment group from each of 6 chickens, after slaughter, jejunum, ileum and duodenum and cecum intestinal contents were collected under sterile operation. 【Result】Gastrointestinal tract Enterobacteriaceae, Lactobacillus, Bifidobacterium and Enterococcus in broilers under immunological stress were analyzed by RT-PCR. The dynamic distribution and changes of the four bacterial species in the broiler's digestive tract were also analyzed systematically. Among the 4 genera of bacteria, changes mainly concentrated in the ileum and cecum, and the Enterobacteriaceae content was 5.03-7.88 copies/mg, Lactobacillus content was 5.48-10.34 copies/mg, Bifidobacterium content was 4.11-7.98 copies/mg, Enterococcus content was 5.61-8.79 copies/mg. The results showed that immune stress makes the four bacterial species in broiler's digestive tract change conspicuously in their genic copy when compared with CV. In which, the changes of the four bacterial species mainly represent in ileum and cecum, while the changes of days by duodenum species is not clear. 【Conclusion】 Immune stress results in the changes of the intestinal flora composition, Lactobacillus and Bifidobacterium are relatively high in NV case and LPS and CYP did not affect intestinal flora changes of chicken in late stage.

Identification and Subcelluar Localization of Bmintegrin αPS3 from Silkworm (Bombyx mori)

TAN Juan, ZHANG Kui, XU Man, CHEN Si-Yuan, CUI Hong-Juan

Scientia Agricultura Sinica. 2013, 46(22): 4808-4815. doi:10.3864/j.issn.0578-1752.2013.22.019

Abstract ( 390 )

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【Objective】Bmintegrin αPS3 was cloned, analyzed, and overexpressed either in Escherichia coli or silkworm cell line to provide a theoretical basis for further studying the function of Bmintegrin αPS3 in silkworm.【Method】The Bmintegrin αPS3 cDNA including the complete ORF sequence was cloned by RACE method. The spatial-temporal expression profile of Bmintegrin αPS3 was investigated by RT-PCR. Bmintegrin αPS3 protein was overexpressed using prokaryotic expression. In addition, Bmintegrin αPS3 was constructed into a transit expression vector, and transfected into silkworm embryonic cell line to analyze the subcellular location of Bmintegrin αPS3.【Result】Bmintegrin αPS3 cDNA consisted of 2 895 bp, and encoded 965 amino acids containing the conserved integrin alpha domains, which was predicted to be a transmembrane protein. Bmintegrin αPS3 protein from E. coli was overexpressed and purified using affinity chromatography, and the purity reached more than 99%. In addition, the overexpressed Bmintegrin αPS3 protein in embryonic cell line was located in cytoplasm and membrane. RT-PCR results showed that Bmintegrin αPS3 mRNA was specifically and continually expressed in silkworm hemocytes.【Conclusion】The Bmintegrin αPS3 cDNA was cloned and its expression patterns in silkworm were analyzed. The subcelluar localization of recombinant Bmintegrin αPS3 protein was investigated in silkworm embryonic cell line. The Bmintegrin αPS3 protein was overexpressed in E. coli and purified for further antibody preparation.

Analysis on the Spatial-Temporal Features and Developing Trend and Effects of Agricultural Geographic Agglomeration in China

DENG Zong-Bing-12, FENG Yong-Gang-2, ZHANG Jun-Liang-2, WANG Ju-2, TIAN Wei-Bo-2

Scientia Agricultura Sinica. 2013, 46(22): 4816-4828. doi:10.3864/j.issn.0578-1752.2013.22.020

Abstract ( 450 )

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【Objective】 This paper is to acquaint the spatial-temporal features, the developing trend of geographic agglomeration and its effects of crop farming. 【Method】 The degree of geographic agglomeration of crop farming was analyzed by comprehensive application of regional Gini index, industry agglomeration rate, regional average agglomeration rate and space correlation index. The agglomeration effects were analyzed by introducing the degree of agglomeration into the C-D production function model. 【Result】 It is found that crop farming generally has significant geographic agglomeration features which differ a lot among different industries. Geographic agglomeration of crop farming tends to be enhanced continuously, which has obvious stage characteristics. The geographic agglomeration features of crop farming are more and more specialized and flaked. The geographic agglomeration of crop farming has a significant positive effect on the industry growth. 【Conclusion】 China should strive to enhance and improve building infrastructure, ameliorate regional management and services, create a favorable external environment for the development of agriculture agglomeration; promoting agricultural specialization and regionalization process should be based on comparative advantages.

Competitive or Complementary? The Adoption Trend of Starch-Based Syrups and Sugar in Food and Beverage Manufacturing

SI Wei-1, ZHU Hai-Yan-12

Scientia Agricultura Sinica. 2013, 46(22): 4829-4836. doi:10.3864/j.issn.0578-1752.2013.22.021

Abstract ( 422 )

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【Objective】 The objective of this paper is to investigate the likely adoption rate of starch-based syrups (SBS) in food and beverage manufacturing, project the demand growing trend of SBS and assess its economic implications for major sweetener industry participants. 【Method】 The logistic growth model that is based on the theory of innovation adoption was emploged to investigate the time pattern of adoption for SBS, and the demand model was used to project the demand for sugar and SBS. 【Result】 China’s sugar and sweetener demand are still at an expanding phase. The market shares of SBS would continue to increase rapidly about ten years with a lower growth rate compared to the previous decade. The growing SBS consumption will decrease the growth rate of demand for sugar, and change the caloric sweetener market structure which is dominated by sugar. The results also show that the decreased manufacturing costs due to the use of SBS should be passed to consumer in lower retail prices. This would make sugar demand more elasticity. The impact account for increasing adoption SBS in food and beverage manufacturing on corn market will be small. 【Conclusion】 The consumption growth of SBS ultimately depends on decreased manufacturing costs due to the use of SBS. Under current market conditions, SBS and sugar are complementary to some extent, rather than simply competitive.

Interspecific Competition Between Encarsia sophia and E. formosa and Their Impacts on Suppression of Trialeurodes vaporariorum

LIU Lin-Zhou-1, DAI Peng-1, 吕Bing-1 , ZANG Lian-Sheng-1, DU Wen-Mei-1, WAN Fang-Hao-2

Scientia Agricultura Sinica. 2013, 46(22): 4837-4841. doi:10.3864/j.issn.0578-1752.2013.22.022

Abstract ( 460 )

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【Objective】The objective of this study is to determine the potential of Encarsia sophia, a heteronomous hyperparasitoid, replacing primary parasitoids that share the same hosts and disrupting whitefly suppression in combination with primary parasitoids E. formosa. 【Method】 In this study, E. sophia and the primary parasitoid, E. formosa were released at ratios of 10﹕0, 8﹕2 , 5﹕5, 2﹕8 and 0﹕10 for controlling the greenhouse whitefly, Trialeurodes vaporariorum, on tomato. After releasing, the population dynamics of E. sophia, E. formosa and T. vaporariorum were investigated every 20 d.【Result】The percentage of E. formosa in total parasitoids was always kept about 50% sixty days after both parasitoids were released at different ratios. Releases of any one parasitoid species only or both of the parasitoids at different ratios provided low levels of whitefly population. Especially, the population of T. vaporariorum was always effectively suppressed with more than 83% whiteflies killed by parasitism and host feeding after E. sophia and E. formosa were released at a ratio of 5﹕5.【Conclusion】The heteronomous hyperparasitoid (E. sophia) can’t replace the primary parasitoid (E. formosa) on T. vaporariorum. The competitive interactions between the two parasitoid species do not reduce the efficiency of whitefly suppression.

Surveys and Analysis of Competition and Displacement Between Two Invasive Species of Leafminer Fly in Hainan Province

WANG Kai-Ge-1, YI Hao-12, LEI Zhong-Ren-1, XIANG Jun-Cheng-1, LIAN Zhen-Min-2

Scientia Agricultura Sinica. 2013, 46(22): 4842-4848. doi:10.3864/j.issn.0578-1752.2013.22.023

Abstract ( 429 )

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【Objective】The objective of this study is to understand the process and the mechanisms of the competition and displacement between the two invasive species of leafminer flies (Liriomyza sativae and L. trifolii) which ecological niches are extremely similar through successive surveys and analysis of their population dynamics in Hainan Province. 【Method】Combining inductive analysis of the surveys of the population dynamics for 8 years in Hainan and the experiments of insecticide resistance, temperature and reproductive interference, the mechanisms of the population replacement between the two species were analyzed.【Result】 The range of the host plants of L. trifolii was broader than that of L. sativae. L. trifolii had distributed across Hainan since it was found in four areas in 2006. The population proportion of L. trifolii had been increased by years since it was firstly found in 2006 in Sanya, Hainan. The surveyed results from 2012 and 2013 indicated that L. trifolii had already become the predominant species in Sanya, Ledong and Lingshui and their population proportions were about 95%.【Conclusion】The competitive advantages of L. trifolii were more significant than that of L. sativae. The stronger ability of reproductive interference and insecticide resistance and the broader range of suitable temperatures and host plants than that of L. sativae were the main factors and the potential suitable establishment areas of L. trifolii was huge in China.

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