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Table of Content

    01 December 2016, Volume 49 Issue 23
    CONTENTS
    Scientia Agricultura Sinica. 2016, 49(23):  0.  doi:10.3864/j.issn.0578-1752.2016.23.000
    Abstract ( 236 )   PDF (330KB) ( 325 )   Save
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    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Field Identification of Drought Tolerance of LOS5 Transgenic Maize
    LIU Cheng, YANG Bing-peng, SUN Bao-cheng, ZHANG Jia-chang, TANG Huai-jun, WANG Tian-yu, ZHANG Deng-feng, XIE Xiao-qing, SHI Yun-su, SONG Yan-chun, YANG Xiao-hong, LI Yu, LI Jian-sheng
    Scientia Agricultura Sinica. 2016, 49(23):  4469-4479.  doi:10.3864/j.issn.0578-1752.2016.23.001
    Abstract ( 598 )   HTML ( 2 )   PDF (451KB) ( 519 )   Save
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    ObjectiveDevelopment of transgenic drought tolerant varieties of maize is an effective way to solve the production problems such as drought disaster and water shortage. The objective of this study was to identify and analyze the effects LOS5 on drought tolerance of maize which were transferred with Arabidopsis thaliana LOS5 gene. And, the transgenic LOS5 maize lines with strongest drought tolerance were screened out for the maize breeding. Method Under the gradient stress of 150, 225, 300, 375, 450 and 600 mm (CK) irrigation amount, the yield and drought tolerance of eight LOS5 transgenic maize lines and their receptor Zheng58 were identified in field. Yield components and other agronomic traits were measured and compared for determining what were the characters most correlated with the enhanced drought tolerance. The transgenic maize lines with significant drought tolerance were screened out for the transgenic breeding application.ResultThe yields and drought tolerances of all 9 maize lines decreased gradually with the decreasing of irrigation amount from 600 to 150 mm. The yields of 8 transgenic maize lines were all significantly higher than that of the receptor Zheng58 in the stress range of 225 mm to 450 mm, and the maximum difference was in the half irrigation (300 mm) of the normal irrigation (600 mm). In the irrigation amount of 300 mm, the drought resistant coefficients of 8 transgenic lines were 0.56 to 0.70, which were significantly higher than that of the receptor Zheng58(0.5), and the drought tolerance then increased by 12% to 40%. The materials in descending order of the drought resistant coefficient were T8920B6, T8920B2, T8920B7, T8920B5, T8920B1, T8920B4, T89B3, B8920B8 and the receptor Zheng58. In the respect of vegetative development, only the leaf color SPAD value of transgenic lines (38.4 to 42.4) were significantly higher than that of the receptor Zheng58 (28.7 to 37.5) in the irrigation of 150 to 225 mm, but the biomass above the ground, plant height and ear site height had no significance among the 9 maize lines. In the development of ear, the ear weight and the ear length of 8 transgenic lines were 42.3 to 61.6 g and 10.9 to 13.1 cm, which were significantly higher than 36.4 to 40.7 g and 8.5 to 11.8 cm of the receptor, respectively, in the irrigation treatments of 150 to 225 mm, but the cob weight and ear diameter were not with significance among the transgenic lines and their receptor. In the grain development, the grain numbers per ear and the grain number per row of the 8 transgenic lines were 86.6 to 182.6 and 8.4 to 15.6, which were significantly higher than 57.3 to 83.2 and 4.9 to 7.1 of the receptor, respectively, but no significant difference was found among the 8 transgenic maize lines and their receptor in the respect of grain rows per ear and 100-grains weight.ConclusionLOS5 had little effect on the vegetable growth of transgenic maize before flowering, but it played a positive role in maintaining of ear length, grain number per row and leaf color of late-filling stage after flowering. And, the ear weight, grain number per ear and yield of transgenic maize were therefore maintained at higher levels when stressed. Drought tolerance of LOS5 transgenic lines was enhanced with different degrees, so field identification and screening is necessary. Stress with a half of normal irrigation can maximize the difference of maize drought tolerance, and was satisfactory for the identification and screening.

    QTL Analysis of Test Weight Dynamic Change in Maize
    XU Meng-meng, QIN Yong-tian, CHEN Yong-qiang, ZHANG Zhan-hui, TANG Ji-hua, FU Zhi-yuan
    Scientia Agricultura Sinica. 2016, 49(23):  4480-4487.  doi:10.3864/j.issn.0578-1752.2016.23.002
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    【Objective】Identifying the QTLs controlling test weight dynamic change during maize grain filling will benefit for gene cloning of the related key QTLs. 【Method】In 2009 and 2010, a set of RIL population with 166 inbred lines derived from hybrid Nongda108 (Xu178 crossed with Huang C) was planted in Zhengzhou and Anyang with randomized block design. The ear samples of recombinant inbred lines with the same silking date were hand-harvested performed by time-course harvesting on 15, 22, 29, 36, 43, and 50 days after pollination for test weight evaluation. Phenotypic analysis was performed using SPSS18.0 software. The 822 pairs of SSR covering the whole maize genome were used to detect the polymorphisms between the two parents. And, 216 SSR markers were selected for linkage map construction. With the linkage map, QTLs responsible for test weight were evaluated by composite interval mapping program in the WinCartQTL 2.5 software. 【Result】Test weight of Nongda108 and its two parents tended to be a slow-fast-slow pattern along with the grain filling process. Genetic factor showed predominant factor on test weight, while environmental factors mainly did their effects during the early and the late stages rather than the peak stage. Between two years, the test weight of DAP22-DAP43 showed a significant difference, but no significant difference at the last stage. The test weight of hybrid Nongda108 was among the two parents exhibiting typical additive effects within different developmental periods. Via time-related QTL analysis, 31 QTLs were detected for test weight of six sampling stages under four environments. These QTLs were located on all the 10 chromosomes of maize genome with exception of chromosomes 4 and 10. And, the 2, 4, 5, 3, 4, 5, 3, and 3 QTLs were distributed on chromosomes 1, 2, 3, 5, 6, 7, 8 and 9, respectively. Among them, additive effect of 13 QTLs was contributed by Huang C (+), while additive effect of another 18 QTLs was contributed by Xu178 (-). The contribution of single QTL varied from 5.9% to 29.7%. qTW2c was detected at DAP22 and DAP36 with a contribution of 11.5% and 14.7% to test weight in Anyang in 2010. qTW3c was detected on DAP29 and DAP36 with a contribution of 22.2% and 14.7% to test weight in Zhengzhou in 2009. 【Conclusion】Two conserved QTLs, qTW2c and qTW3c, were repeatedly detected at two sampling stages with more than 10% contribution to test weight. These two major QTLs are very important for cloning gene controlling test weight dynamic change.
    Genetic Linkage Map Construction Based on EST-SSR and Analysis of QTLs for Starch Content in Sweetpotato (Ipomoea batatas (L.) lam.)
    TANG Dao-bin, ZHANG Kai, Lü Chang-wen, XIE De-bin, FU Ti-hua, WANG Ji-chun
    Scientia Agricultura Sinica. 2016, 49(23):  4488-4506.  doi:10.3864/j.issn.0578-1752.2016.23.003
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    ObjectiveTo provide a theoretical and practical basis for utilization of germplasm resources of sweetpotato with high starch content and molecular marker-assisted selection in sweetpotato, a genetic linkage map was constructed by using molecular markers, and quantitative trait loci (QTLs) associated with starch content were identified. Method A F1 hybrid population was developed from a cross between Wanshu 5, a sweetpotato cultivar with high starch content, as the female parent, and Shangqiu 52-7, a cultivar with low starch content, as the male parent. By using EST-SSR markers and the software of JoinMap4.0, the molecular genetic linkage maps of both parents were constructed based on the “double pseudo testcross” strategy, respectively. And QTLs for starch content trait were identified using composite interval mapping method based on the phenotypic data of F1 population in two years (2012 and 2013).ResultAmong the 1 679 EST-SSR primer pairs primarily tested in F1 lines, 1 045 polymorphic primer pairs were selected to screen in the population lines, and 1 418 polymorphic loci were obtained. The genetic linkage relationship of polymorphic loci were analyzed with the LOD5.0 as a threshold, and the genetic linkage map of female and male parents were constructed respectively. A total of 74 linkage groups for female parent were constructed based on 642 polymorphic loci, of which 215 (33.5%) loci were placed on the genetic linkage map. The number of markers in each linkage group ranged from 2 to 11. The length of linkage groups ranged from 2.0 to 156.8 cM, and the linkage map covered a total length of  3 826.07 cM, with an average distance between markers of 17.80 cM. A genetic linkage map for male Shangqiu52-7 was constructed by using 776 polymorphic loci, of which 250 (32.5%) loci distributed on 80 linkage groups. The number of markers in each linkage group ranged from 2 to 24. The length of linkage groups ranged from 2.0 to 156.8 cM, and the linkage map covered a whole length of 3 955.0 cM with an average interval of 15.7 cM between markers. Using QTL analysis software Map QTL 5.0 and Interval Mapping method, QTLs for starch content were identified based on the starch content measured in two years (2012 and 2013) and two environments. Seventeen QTLs for starch content were detected, explaining 8.4%-40.5% of the phenotypic variation. Three QTLs qWsc-1, qWsc-2, and qWsc-3 mapped on the linkage group of female Wanshu 5 were detected under two environments. Fourteen QTLs were detected on the linkage group of male Shangqiu52-7, of which qSsc-1, qSsc-2, qSsc-3, qSsc-4, qSsc-8, qSsc-10, qSsc-11, qSsc-12 were detected under two environments, and qSsc-5, qSsc-6, qSsc-7, qSsc-9, qSsc-13, qSsc-14 were only detected under one environment. Mark GDAAS 0603 was detected simultaneously on the linkage groups of both parents and two environments. Those QTLs for starch content could be used in molecular marker-assisted selection of sweetpotato. ConclusionTwo genetic linkage map were constructed based on EST-SSR markers, and 17 QTLs for starch content of sweetpotato were identified. Results of the study has enriched the types of molecular markers used for genetic linkage map construction in sweetpotato.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Water Use Efficiency of Winter Wheat Based on AquaCrop Model in Beijing
    XING Hui-min, XU Xin-gang, FENG Hai-kuan, LI Zhen-hai, YANG Fu-qin, YANG Gui-jun, HE Peng, CHEN Zhao-xia
    Scientia Agricultura Sinica. 2016, 49(23):  4507-4519.  doi:10.3864/j.issn.0578-1752.2016.23.004
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    【Objective】 Water use efficiency (WUE) is an important index of agricultural water management and decision. Water shortage is serious and winter wheat is the main water consuming plant, so this study analysed the winter wheat WUE to provide information and support for decision makers for winter wheat water saving irrigation and yield balance in Beijing.【Method】 The response characteristics of yield and WUE of winter wheat under different rainfall year types and irrigation treatments were analyzed using the experimental data collected in 2011-2012, 2012-2013 and 2013-2014 to simulate and validate the model under different years and irrigations and then calibrate the AquaCrop model in National Precision Agriculture Research and Demonstration base. The daily precipitations of winter wheat growing period were collected from 2004 to 2014 in Beijing, and then using the Pearson-III distribution to divide three precipitation types, wet year (2012-2013), normal year (2009-2010) and dry year (2005-2006). The response characteristics of grain yield and WUE were simulated under three precipitation types and 14 irrigation scenarios using the AquaCrop model. 【Result】 The correlation coefficient (R2), root mean square error (RMSE) and consistency (d ) of simulated and measured values of yield were 0.99, 0.3 t·hm-2 and 0.99, respectively. The simulated and measured values of winter wheat grain WUE were 1.72 kg·m-3 in 2011-2012 growth period, 1.67 kg·m-3 in 2012-2013 growth period and 1.27, 1.74 and 1.64 kg·m-3 under rainfall, normal irrigation and over-irrigation in 2013-2014. The highest WUE was under normal irrigation, the second was under over-irrigation and the lowest was under rainfall. The response characters of winter wheat grain yield and WUE under three precipitation types were simulated by AquaCrop model. The irrigations were 35 and 50 mm when yield and WUE were the highest in wet year, 35 and 40 mm in normal year and 65 mm in dry year, respectively.【Conclusion】 The results of the study show that the AquaCrop model well predicted the winter wheat grain yield and WUE under different years and irrigations in Beijing. The grain yield and WUE increased when irrigations increased, began to decrease when reached the maximum values. The plants could improve the WUE through their adaptation when water is in shortage, the WUE decreased with rainfall or irrigation increasing, therefore the ranking of the WUE was dry year, normal year and wet year under three precipitation types. Therefore both the yield and WUE must be considered in development of the irrigation strategies for winter wheat. The AquaCrop model can be used to provide guidance for the field irrigation and decision making of winter wheat in Beijing region. In this study, the grain yield and WUE were only simulated under wet year, normal year and dry year in over-wintering period, regreen period, stem elongation period, blooming period and grain filing period, the effects of different irrigations on grain yield and WUE were not considered in different periods, which need to be further studied and verified.
    Temporal and Spatial Variations of Leaf Shape Coefficients of Summer Maize
    ZHOU Yuan-gang, LI Hua-long, JIANG Teng-cong, DOU Zi-he, LIU Jian, WU Shu-fang, FENG Hao, ZHANG Ti-bin, HE Jian-qiang
    Scientia Agricultura Sinica. 2016, 49(23):  4520-4530.  doi:10.3864/j.issn.0578-1752.2016.23.005
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    【Objective】Leaf shape coefficient (α) provides a simple and fast way for the measurement of crop leaf area and LAI (leaf area index) in field. However, the selection of values of this coefficient was very arbitrary and there was no unique standard to follow. In addition, this coefficient was usually considered as a constant regardless of its variations through the whole lifetime of a given crop. 【Method】 In this study, the temporal and spatial variations of α values of summer maize were investigated through a field experiment conducted from June to October in 2015. A total of six maize cultivars with different properties of ripening were involved. The whole growth season of maize was divided into six different stages, i.e. trefoil, jointing, heading, flowering, silking, and maturity. Maize plants were randomly sampled every six days and all leaves were cut off and measured for their length, width, and area with a digital leaf area scanner. Then, α value was calculated for each leaf. The variations of α values were analyzed for different growth stages and among different leaf positions within a single maize plant. Finally, five different models of leave area estimation, which belong to the linear, quadratic, and logarithmic types, were established to estimate the area of each maize leaf. Three different statistics of RMSE (root mean square error), RRMSE (relative root mean square error), and ARE (absolute relative error) were used to represent the estimation accuracy. 【Result】 Based on linear regression analysis between leaf areas and products of leaf length and width of 760 leaf samples of six different maize cultivars, the general average value of &alpha was about 0.78. Then, when estimating maize leaf areas with the model of LA=0.78×L×W, the relative root mean square error (RRMSE) and absolute relative error (ARE) were 9.50% and 6.96%, respectively. The accuracy was the highest among the five different models investigated for the estimation of maize leaf area. The results showed that α value ranged from 0.72 to 0.87 and varied at different growth stages. It increased with fluctuations from trefoil to flowering stage, and then decreased. At flowering stage, the α value was significantly different from other before-flowering stages, while showed no significant difference at silking and maturity stages. For different ripening cultivars, α value only showed a significant difference at flowering and silking stages The α value varied for different leaf shapes in the whole growth season. From the trefoil to before-jointing stage, α value of wide-short leaves was higher than that of thin-long leaves. From then on, α value of wide-short leaves became lower than that of thin-long ones. Within a single maize plant, α values varied remarkably for leaves at different positions. At the flowering, silking, and maturity stages, α values were higher at top and bottom than in the middle. The average value of three-ear-leaves was 0.78 and the standard deviation was less than 0.05. However, α value was about 0.87 for leaves at the top and bottom of maize canopy, with standard deviations from 0.03 to 0.10. The α values at different leaf positions only showed significant difference at flowering, silking, and maturity stages. 【Conclusion】 When estimating maize leaf areas with the model of LA=0.78×L×W, the accuracy was the highest among the five different models investigated. The leaf shape coefficient of 0.78 improved the estimation accuracy of maize leaf area by about 3.86%, when compared with the estimation resulted from a coefficient of 0.75. In general, it is better to adopt various α values at different growth stages and different leaf positions so as to improve the accuracy of simulation and prediction of leaf area of summer maize.
    Effects of Super-High-Yield Soybean Cultivars as Rootstock on Some Physiological and Yield Traits of Cultivars Released in Different Decades
    TENG Fei, LI Sheng-you, RAO De-min, YAO Xing-dong, ZHANG Hui-jun, AO Xue, WANG Hai-ying, Steven St.Martin, XIE Fu-ti
    Scientia Agricultura Sinica. 2016, 49(23):  4531-4543.  doi:10.3864/j.issn.0578-1752.2016.23.006
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    【Objective】 The objective of this study is to determine the effects of super-high-yield soybean cultivars as rootstocks on some physiological traits of cultivars released in different decades, in order to reveal the important role of root improvement in soybean breeding. 【Method】 The 11 cultivars released in different decades by Liaoning, China and Ohio, USA, having the common ancestors Amsoy and Williams, were used to graft onto the rootstocks of super-high-yield cultivars Liaodou 14 and Zhonghuang 35 by cleft grafting method at cotyledon stage, respectively. The 11 cultivars released in different decades treated with self-grafting and non-grafting were used as the control. Under the conditions of pot experiment, the photosynthetic rate, stomatal conductance, transpiration rate and leaf greenness values were determined at R1, R2, R4, R5, and R6 stages, the seeds per plant, 100-seed weight, seed weight per plant were determined at mature stage and after harvest to test the plants. 【Result】 The root vigor of soybean cultivars at different growth stages were characterized by Liaodou 14 > Zhonghuang 35 > 11 cultivars released in different decades, at R5 and R6 were at significant or extremely significant level. The photosynthetic rate, stomatal conductance, transpiration rate and leaf greenness values showed a single peak curve in the whole growth stages. The photosynthetic rate, stomatal conductance, transpiration rate reached peak value at R2 stage, and the leaf greenness values reached peak value at R4 stage, and then began to decline. Cultivars released in different decades had similar curves between non-grafted and self-grafted plants. However, the decline of the curve became slower after being grafted onto the rootstocks of super-high-yield cultivars Liaodou 14 and Zhonghuang 35 than that of self-grafted plants. At R5 to R6 stages, the daily decrease rates of non-grafted and self-grafted plants for photosynthetic rate, stomatal conductance, transpiration rate and leaf greenness were by 1.43%, 3.67%, 3.38% and 0.82%, respectively, but those of cultivars grafted onto Liaodou 14 and Zhonghuang 35 rootstocks were by 1.11%, 3.26%, 2.05% and 0.48%, respectively. The seeds per plant, 100-seed weight, seed weight per plant of cultivars released in different decades had no significant difference between self-grafted and non-grafted plants. At mature stage, the seeds per plant, 100-seed weight and yield per plant of cultivars released in different decades were increased significantly as grafted them onto super-high-yield cultivars Liaodou 14 or Zhonghuang 35 as rootstocks, by 2.82%, 9.36% and 10.60%, respectively, and the increase of current cultivars was greater than middle cultivars and common parents. 【Conclusion】 The roots of super-high-yield cultivars Liaodou 14 and Zhonghuang 35 have higher physiological activity at reproductive stage. Cultivars grafted onto the rootstock of super-high-yield cultivar could reduce significantly the rate of decline in photosynthetic rate, stomatal conductance, transpiration rate and leaf greenness value, and improve the photosynthetic ability, and thus increasing grain yield. Therefore, more attention should be pay to the genetic improvement of root in the future breeding work.
    PLANT PROTECTION
    Cloning and Expression Analysis of Cytochrome C Oxidase Subunit Ⅲ from Sitophilus zeamais
    HOU Chang-liang, WANG Jing-bo, LI Yong-qiang, WU Hua, MA Zhi-qing, FENG Jun-tao, ZHANG Xing
    Scientia Agricultura Sinica. 2016, 49(23):  4544-4554.  doi:10.3864/j.issn.0578-1752.2016.23.007
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    【Objective】The objective of this study is to clone full-length cDNA of cytochrome C oxidase subunit Ⅲ (COXⅢ), which is a possible action site of allyl isothiocyanate (AITC), then analyze the sequence and prokaryotic expression.【Method】Firstly, finding the known part of the gene of COXⅢ in the NCBI, the full sequence of Sitophilus zeamais COXⅢ was cloned by RT-PCR and RACE techniques. The sequences of COXⅢ, phylogenetic relationship and properties of S. zeamais COXⅢ, the three-dimensional structure were analyzed by a variety of bioinformatics softwares such as DNAMAN8.0, MEGA5.1, GENEDOC, ProtParam and TargetP 1.1 Server et al. Using pEASY-Blunt E1, pET28a, pET30a, PET32a and pET42a as a fused expression vector, a recombinant plasmid pEASY-Blunt E1-COXⅢ, pET-28a-COXⅢ, pET30a-COXⅢ, pET32a-COXⅢ, pET42a-COXⅢ were constructed. Then inducing its expression in Escherichia coli BL21 (DE3) with IPTG at different concentrations, times and temperatures. SDS-PAGE was used to detect the fusion protein expression and the protein expression was verified by Western blot.【Result】The full-length sequences of COXⅢ from S. zeamais was obtained, and its ORF is 792 bp encoding 263 amino acid residues, the predicted molecular weight is 30 938.1 Da, isoelectric point pI is 6.51, the prediction formula is C1492H2154N338O362S10, the unstable factor is 34.49, and the total average hydrophilic coefficient is 0.492. It is a stable hydrophobic protein. Phylogenetic tree showed that S. zeamais had the closest evolutionary relationship with S. oryzae at the amino acid level. Prokaryotic expression results showed that efficient expression of COXⅢ protein could be realized after induced with 1 mmol·L-1 IPTG in E. coil BL21 (DE3) for 24 h at 18℃. Solubility analysis showed that the fused protein mainly existed as inclusion bodies. Western blot confirmed that the molecular weight of the recombinant COXⅢ is about 62 kD, consistent with the predicted result.【Conclusion】The full-length sequences of COXⅢ from S. zeamais was successfully obtained, and the prokaryotic expression of its encoding protein was achieved, which will lay a foundation for further function research of S. zeamais COXⅢ, and provide a theoretical basis for the research of the mechanism of the action on controlling S. zeamais by using isothiocyanates compound.
    Expression and Molecular Simulation of Alkaline Phosphatase Receptor of Plutella xyllostella
    ZHANG Xiao, HU Xiao-dan, ZHONG Jian-feng, WU Ai-hua, XU Chong-xin, LIU Yuan, ZHANG Cun-zheng, XIE Ya-jing, LIU Xian-jin
    Scientia Agricultura Sinica. 2016, 49(23):  4555-4565.  doi:10.3864/j.issn.0578-1752.2016.23.008
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    【Objective】The objective of this study is to confirm the binding ability of membrane-bound alkaline phosphatase (mALP) of Plutella xyllostella with Cry1Ac toxin using prokaryotic expression and Ligand blot, and to predict toxin-receptor binding region and key amino acid binding sites (hot-spots) employed by homology modeling and docking study of Cry1Ac-mALP binding mode. It will provide a basis for the study of toxin-receptor interaction mechanism and molecular modification to enhance the activity of Cry toxin. 【Method】 The mALP of P. xyllostella full-length primers were designed and amplified by PCR. The restricted products of mALP and pET-26b (+) were ligated by T4 DNA ligase after the dual-enzyme digestion procedures. The recombinant pET-26b-mALP vectors were transferred into the Trans1-T1 phage resistant chemically competent cells, then picked clones were analyzed by PCR amplification, dual-enzyme digestion and sanger sequencing. The positive recombinant vectors (anchoring the corrected mALP gene) were transferred into the E. coli BL21 (DE3) competent cells for prokaryotic expression. The inducible expression products of mALP were transformed onto PVDF membrane. Preparation of mALP and binding activity of Cry1Ac with mALP were verified through Western blot and Ligand blot, respectively. Three-dimensional structure of mALP was predicted by homology modeling, molecular dynamics simulation and model evaluation. The toxin-receptor docking complexes were generated by using the PatchDocK and FireDock web-servers with molecular dynamics simulations. The toxin-receptor complex was analyzed to determine the interaction region and the amino acid binding sites, key amino acid residues involved in Cry toxin and ALP receptor by computer-aided alanine mutation scanning tests. 【Result】P. xyllostella mALP gene was successfully amplified, followed with the prokaryotic expression of mALP receptor protein. Binding of Cry1Ac toxin with prepared mALP protein was verified. The three-dimensional structure of mALP was successfully obtained by homology modeling, then the Cry toxin-ALP complex was determined. By the changed solvent accessible surface areas calculation and Ligplot analysis, the results showed that the domain II and domain III of Cry toxin were involved in binding to receptor, and Cry toxin and ALP were interacted mainly depending on hydrophobic and hydrogen bonding patterns. Finally, through the computer-aided alanine mutation scanning hot residues analysis, there were three key amino acid residues (376ASN, 443SER and 486SER) from Cry toxin and four key amino acid residues (452ARG, 499THR, 502TYR and 513TYR) from ALP were participated in the interaction of toxin-receptor complex, respectively. 【Conclusion】It can be determined that the mALP receptor also has the ability to bind Cry1Ac toxin by prokaryotic expression, the three-dimensional structure of mALP was predicted and the toxin-receptor binding model was studied using molecular simulation.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Assessment of Soil Fertility and Its Spatial Variability Based on Small Scale in the Gravel Mulched Field of Ningxia
    WANG You-qi, BAI Yi-ru, ZHAO Yun-peng
    Scientia Agricultura Sinica. 2016, 49(23):  4566-4575.  doi:10.3864/j.issn.0578-1752.2016.23.009
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    【Objective】The gravel mulched field is a unique method of tillage in arid and semi-arid areas of northwest under the natural condition with severe weather, poor soil and terrain condition, through the long-term production practice, continuous summarization and innovation. Preservation and improvement of soil fertility quality is a foundation for maintaining land productivity, preventing land degradation in the gravel mulched field. 【Method】A total of 110 soil samples (0-20 cm) were collected in a grid of 10 m×10 m. Soil indices were selected as evaluating factors including soil organic carbon, total N, total P, total K, available P, available K, pH and electrical conductivity. Correlation coefficient method was used to determine the weight value of soil fertility index. The curve type and turning point value of membership function were defined by soil characteristics and crop varieties. Then membership degree and weight value were combined to calculate the soil integrated fertility index (IFI). The geostatistics method was used for building the best semi variance function models, and analyzing spatial variation and distribution pattern of soil fertility. The evaluation of classification of soil fertility could be studied by the soil IFI and its spatial distributions. 【Result】The classical statistics indicated that the mean value of pH was 9.15 showed alkalinity, and the coefficient of variation was 0.01 showed medium variation. The coefficient of variations for other fertility indexes ranged from 0.10 to 0.72 showed moderate variation. The range of soil characteristics showed an obvious difference at field scale. The soil IFI in the study area was 0.18-0.59 with the average value of 0.34. The coefficients of nugget for fertility indexes were less than 25%, which demonstrated strong spatial dependence. The ordinary Kriging interpolation map indicated that the soil organic carbon distributed evenly with the mean value of 1.5-2.5 g·kg-1, which had island high levels in the southern study area. The electrical conductivity also distributed evenly with the mean value of 100-300 μs·cm-1, which had high levels in the northern study area. Total N, total P and total K had low levels with plaque distribution. The available P and available K were higher in the northern study area. The pH value also distributed patchily. The land of Grade Ⅲ indicated medium fertility condition and distributed most widely in the study area, covered 56% of the total area. The land of Grade Ⅱ indicated fecund fertility condition and represented the main type in the south of the land, covered 25% of the total area. The land of Grade Ⅳ accounted for 18% of the total area, which indicated poor fertility condition. The soil property indexes and IFI value demonstrated that soil fertility level was relatively low in the gravel mulched field. 【Conclusion】The spatial distribution patterns of soil index and IFI were complicated and had no regularity with patch distribution at field scale. According to the soil index and IFI, the levels of soil fertility were low in the gravel mulched field. The soil organic carbon and total N were the main limiting factors restraining soil fertility. The methods of increasing organic fertilizers, crop rotation and fallow were practicable and necessary for fertilizing soil, increasing yields and improving crop quality.
    Mitigative Effect of Biochar on Aluminum Toxicity of Acid Soil and the Potential Mechanism
    YING Jie-guan, LIN Qing-yi, ZHANG Meng-yang, HUANG Yi, PENG Shu-ang, JIANG Cun-cang
    Scientia Agricultura Sinica. 2016, 49(23):  4576-4583.  doi:10.3864/j.issn.0578-1752.2016.23.010
    Abstract ( 524 )   HTML ( 5 )   PDF (641KB) ( 502 )   Save
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    【Objective】Aluminum toxicity of acid soil is one of the hot topics in soil science research, and biochar with some bio-characters give a new method to solve this problem. The objective of this experiment is mainly to clarify the mitigative effect of biochar on aluminum toxicity of acid soil and the potential mechanism. 【Method】 An experiment including CK (0C+0Al), C (2%C), Al (1 mmol·L-1Al), and C+Al (2%C+1 mmol·L-1Al) treatments indoor planting pakchoi in pots added with biochar and exogenous aluminum was conducted. The effects of biochar on different active aluminum and the growth of crop of aluminum enrichment red soil were analyzed. 【Result】 The results showed that aluminum toxicity significantly decreased the emergence rate of pakchoi. The growth of pakchoi restrained significantly and the biomass of pakchoi decreased under aluminum toxicity, meanwhile, the Al content of the pakchoi increased significantly under aluminum toxicity. However, the growth of pakchoi improved significantly and the Al content reduced under biochar addition, and the aluminum content of C+Al treatment pakchoi reduced by 89.4% compared with Al treatment. Aluminum toxicity decreased the pH of the red soil, the pH of Al treatment decreased by 0.36 compared with CK treatment, but the pH of the red soil increased significantly under biochar addition, the pH of C+Al treatment increased by 0.62 compared with Al treatment. The active aluminum of Al treatment was 276.4 mg·kg-1 more than CK treatment. Application of biochar decreased the active aluminum of red soil, and the active aluminum in C+Al treatment decreased by 14.9% compared with Al treatment. Besides, the exchangeable Al3+ of Al treatment increased by 23.1% compared with CK treatment, but the exchangeable Al3+ of C+Al treatment decreased by 46.5% compared with Al treatment. The main active aluminum form of CK and Al treatment soil was the exchangeable Al3+ that had biological toxicity, the main active aluminum form of C and C+Al treatment soil was dimer hydroxy aluminum ion and colloid Al(OH)30. 【Conclusion】 Exogenous aluminum had a strong stimulating effect on the addition of active aluminum of red soil. Exchangeable Al3+ which has biological toxicity increased significantly under aluminum toxicity. Besides, adding exogenous aluminum can decrease the pH and stimulate the poison, restrain the growth of the crops. Although application of biochar can increase the pH and ease the aluminum toxicity of red soil, its effect on different 4 forms of active aluminum has a quite large difference. Application of biochar can reduce the poison by decreasing the exchangeable Al3+ which has biological toxicity, and thus improve the growth of crop.
    HORTICULTURE
    Isolation and Expression Analysis of a Vacuolar Glucose Transporter Gene MdVGT1 in Apple
    XU Hai-feng, LIU Jing-xuan, WANG Yi-cheng, ZUO Wei-fang, QU Chang-zhi, WANG De-yun, ZHANG Jing, JIANG Sheng-hui, WANG Nan, CHEN Xue-sen
    Scientia Agricultura Sinica. 2016, 49(23):  4584-4592.  doi:10.3864/j.issn.0578-1752.2016.23.011
    Abstract ( 546 )   HTML ( 9 )   PDF (825KB) ( 850 )   Save
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    【Objective】 In order to develop the theory and breeding technology of functional apple, the bioinformatics, the expression level and the function in the sugar metabolism several aspects of vacuolar glucose transporter gene MdVGT1 in ‘Hongcui No.1’ of Malus sieversii f. neidzwetzkyana F1 population were studied. 【Method】 The MdVGT1 in ‘Hongcui No.1’ was cloned and the bioinformation of it was analyzed. The expression level of MdVGT1 in different tissues and different development stages was studied by the qRT-PCR and the expression of MdVGT1 induced by glucose in tissue culture seedlings of ‘gala’ was also analyzed. Meanwhile, the interaction between MdVGT1 and MdTMT1 was verified by yeast two-hybrid system and the recombinant protein by the prokaryotic induction technology was obtained. 【Result】 The full length of MdVGT1 in ‘Hongcui No.1’ was cloned, and the gene was 1 506 bp which encoded 501 amino acids. It was predicted that the molecular mass of this protein was 53.16 kD, and pI was 5.92. Furthermore, it was inferred that the gene includes 14 exons and 13 introns, and is located on the chromosome 1 of the apple genome. A phylogenetic tree indicated that MdVGT1, AtVGT1, VvVGT1 and CsVGT1 are located in the same evolutionary branch. Analysis of functional domain showed that MdVGT1 contains 12 transmembrane regions. MdVGT1 has the higher expression level in flowers, leaves and young fruits of apples, and its expression has a significant correlation with the content of glucose during the development stage of fruits. The promoter of MdVGT1 contains several typical cis-acting elements, including defense responsive elements, sugar signaling responsive elements and phytohormone responsive elements, and the expression of MdVGT1 in tissue culture seedlings of ‘gala’ was significantly increased after glucose treatment a week later. The yeast two hybrid experiments showed that the MdVGT1 could interact with MdTMT1 in vitro. In addition, the recombinant protein of MdVGT1 was obtained by the prokaryotic induction technology. 【Conclusion】The vacuolar glucose transporter gene MdVGT1 in ‘Hongcui No.1’ apple was cloned, and it was found that it could interact with MdTMT1 to transport glucose into the vacuole membrane.
    Cloning and Expression Analysis of Ten MADS-box Genes in Peach (Prunus persica var. nectarina ‘Luxing’)
    LI Hui-feng, JIA Hou-zhen, DONG Qing-long, RAN Kun, WANG Hong-wei
    Scientia Agricultura Sinica. 2016, 49(23):  4593-4605.  doi:10.3864/j.issn.0578-1752.2016.23.012
    Abstract ( 424 )   HTML ( 4 )   PDF (940KB) ( 609 )   Save
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    【Objective】The aim of this study is to characterize the novel peach (Prunus persica var. nectarina ‘Luxing’) MADS-box genes (PpMADSs) involved in regulation of vegetative and reproductive growth. The transcriptional levels of PpMADSs in different tissues were determined to provide a basis for studying the related function of PpMADSs in flower development, fruit development and ripening.【Method】The full-length cDNA sequences of PpMADSs form ‘Luxing’ peach were isolated by homologous alignment and RT-PCR confirmation, the obtained cDNA sequences and the deduced amino acid sequences were analyzed with bioinformatics methods; the expression levels of PpMADSs were detected in stem, leaf, sepal, ovary, stamen, petal, 7 stages of flower development and 5 stages of fruit development using RT-PCR.【Result】The sequencing results showed that ten cDNAs (designated as PpMADS11, 12, 19, 20, 21, 22, 28, 29, 30 and 31; GenBank accession No. KU559577, KU559578, KU559585, KU559586, KU559587, KU559588, KU559594, KU559595, KU559596 and KU559597) contained open reading frame (ORF) of 522, 279, 1 065, 828, 723, 600, 636, 534, 750 and 480 bp, respectively. The results of phylogenetic analysis revealed that PpMADS11, 12, and 19 belong to AP3, AGL17 and MIKC* subgroups, respectively; and PpMADS20, 21 and 22 belong to Mα group; PpMADS28, 29, 30 and 31 belong to Mγ group. The results of subcellular localization prediction showed that all PpMADS proteins were located in the nucleus. The results of promoter analysis indicated that there were multiple putative cis-acting elements involved in light responsiveness, defense and stress responsiveness, MYB binding site was involved in drought-inducibility, heat stress responsiveness, low-temperature responsiveness, fungal elicitor responsive element, wound-responsive element, anaerobic induction element, gibberellin-responsive element, auxin-responsive element, MeJA-responsiveness, abscisic acid responsiveness, salicylic acid responsiveness and ethylene-responsive element. Semi RT-PCR and qRT-PCR results showed that PpMADS11was expressed in stem, leaf, sepal, ovary, stamen, petal and during flower and fruit development. PpMADS12 was expressed in stem, leaf, sepal, ovary, stamen, petal and during flower development. PpMADS19 was expressed in sepal, stamen, petal and during flower development ( except bud stage). All members in Mα and Mγ groups were expressed in stem, leaf, sepal, ovary, stamen, petal and during flower development, some members were expressed during fruit development.【Conclusion】These results indicated that ten PpMADS genes have crucial regulatory roles in ‘Luxing’ peach vegetative growth, flower and fruit development processes.
    STORAGE·FRESH-KEEPING·PROCESSING
    Effect of Extrusion on Digestion Properties and Volatile Compounds in Germinated Brown Rice Compounded of Flammulina velutipes Flour
    FANG Yong, WANG Hong-pan, PEI Fei, MA Ning, TANG Xiao-zhi, YANG Wen-jian, HU Qiu-hui
    Scientia Agricultura Sinica. 2016, 49(23):  4606-4618.  doi:10.3864/j.issn.0578-1752.2016.23.013
    Abstract ( 425 )   HTML ( 1 )   PDF (4413KB) ( 587 )   Save
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    【Objective】The objective of this study was to investigate the effect of extrusion process on digestion properties and volatile compounds in germinated brown rice compounded of Flammulina velutipes flour through the changes of starch, protein and volatile compounds, thus providing references for the analysis of extrudant digestion properties and flavor.【Method】 In order to study the effects of extrusion on digestion properties and flavor compounds, germinated brown rice compounded of Flammulina velutipes was extruded to investigate the particle microstructure of starch and protein, the protein molecular weight and protein secondary structure, the digestibility of starch and protein, and the volatile compounds using scanning electron microscope, SDS-PAGE electrophoresis, infrared spectroscopy, in vitro digestion method and gas chromatography-mass spectrography.【Result】The results showed that the granule shapes of extruded starch and protein were changed into irregular shape compared with the un-extruded compounded powder. Moreover, the starch content and protein content in germinated brown rice compounded of F. velutipes extrudant decreased by 8.26% and 1.00%, respectively. However, the reduced sugar content, the soluble sugar content, in vitro starch digestibility and in vitro protein digestibility increased by 1.35 times, 18.45%, 30.68% and 25.57%, respectively. The protein molecular weight mainly distributed in 50, 36, 33, 22, 19 and 13 kD in un-extruded compounded powder. However, the 50 kD molecular weight disappeared and the 36 kD molecular weight had no change in germinated brown rice compounded of F. velutipes extrudant. The infrared spectra of protein in extruded were similar to that of un-extruded powder, but the absorption peak intensity had significant difference between 1 645 and 1 544 cm-1. The contents of aldehydes, alcohols, pyrazines and ketones compounds in germinated brown rice compounded of F. velutipes extrudant were increased by 18.93%, 44.17%, 77.64% and 1.56 times, respectively. The aldehydes and alcohols compounds in extruded products of germinated brown rice showed a decline trend compared with un-extruded germinated brown rice. However, the increasing trend to that of the extruded products of germinated brown rice compounded of F. velutipes was exhibited, indicating the flavor enhancement of F. velutipes.【Conclusion】Extrusion technology could improve the digestion characteristics of germinated brown rice compounded of F. velutipes extrudant. Meanwhile, the mixture of F. velutipes could enhance the flavor of the product.
    Multiplex Enrichment Quantitative PCR Assays for the Detection of Vibrio alginolyticus, Vibrio parahaemolyticus, Vibrio vulnificus and Vibrio cholerae
    WEI Shuang, WANG Tian-jie, LONG Yang, ZHOU Guang-biao, LIN Chun-gui, HUANG Shuai, WU Xi-yang
    Scientia Agricultura Sinica. 2016, 49(23):  4619-4627.  doi:10.3864/j.issn.0578-1752.2016.23.014
    Abstract ( 517 )   HTML ( 2 )   PDF (1388KB) ( 362 )   Save
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    【Objective】Vibrio parahaemolyticus, V. alginolyticus, V. cholerae and V. vulnificus have been recognized as the important foodborne pathogens causing human disease. It is important to establish a detection method to identify these 4 foodborne pathogenic Vibrio species in order to ensure food safety. To develop a multiplex enrichment quantitative PCR (ME-qPCR) assay that can simultaneously detect Vibrio parahaemolyticus, V. alginolyticus, V. cholerae and V. vulnificus in the presence of an internal amplification control (IAC), to make it possible for researchers and technical staff achieve simple detection of these 4 foodborne pathogens. 【Method】 Inner and outer species-specific PCR primers were designed based on gyrB gene for V. alginolyticus, collagenase gene for V. parahaemolyticus, vvhA gene for V. vulnificus and ompW gene for V. cholerae, 16S rRNA gene of bacteria as IAC primers was used to indicate false-negative results. All inner and outer primers were used in first round multiplex enrichment PCR with a small number of cycles so as to avoid competition between amplicons. Each gene has 4 types of probable products which can be the templates for the next nested real-time PCR. This can enrich the target genes from the genomic DNA template successfully. The reaction product was then diluted and analyzed individual real-time PCRs using inner primers. The results were analysed by amplification curve and melting curve. ME-qPCR method was developed after optimization of the first round multiplex enrichment PCR cycle numbers (10, 15 and 20 cycles). The specificity of ME-qPCR was validated by 14 bacteria standard strains. The sensitivity and quantitative capability of ME-qPCR were tested by using 10-fold serially diluted genomic DNA of 4 Vibrio strains. Strains as templates, and then, the ME-qPCR was used to detect 69 suspicious Vibrio strains and the results were compared with physiological and biochemical experiments. 【Result】 Fifteen cycles were determined to use in first round multiplex enrichment PCR in this study finally. The results showed that the ME-qPCR assay was rapid, high-throughput, sensitive and specificand the existence of IAC could successfully eliminate false-negative results. The sensitivity of ME-qPCR was 0.001ng per reaction, about 10 times higher than real-time PCR. The ME-qPCR was validated with 69 suspicious Vibrio strains. The result showed that 27 green bacteria colonies were V. parahaemolyticus, 22 yellow bacteria colonies were V. alginolyticus, 1 yellow bacteria colony was V. vulnificus, and V. cholerae was not detected. The results are consistent with physiological and biochemical experiments. 【Conclusion】 The ME-qPCR assay is specific, stable and reliable for the detection of V. parahaemolyticus, V. alginolyticus, V. cholerae and V. vulnificus. It’s sensitivity is high, and can effectively indicate the false negative of PCR reaction, the results without gel electrophoresis, and is suitable for the rapid screening of 4 common pathogenic vibrio. in food
    ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT
    Breeding of Disease Resistant Dwarf Chickens by Phagocytic Ability
    MA Hui, HAN Hong-bing, NING Zhong-hua, LIAN Zheng-xing
    Scientia Agricultura Sinica. 2016, 49(23):  4628-4637.  doi:10.3864/j.issn.0578-1752.2016.23.015
    Abstract ( 327 )   HTML ( 1 )   PDF (940KB) ( 339 )   Save
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    【Objective】 Monocytes-macrophages play an important role in the immune system. The effect of selection for monocytes-macrophages phagocytosis on infectious bronchitis virus (IBV) resistance in generation 1 (G1) of dwarf chickens was studied. 【Method】 The phagocytic index (PI) of 500 dwarf chickens (400 hens and 100 cocks) of generation 0 (G0) was tested at 290 d of age, and then the chickens were divided into high and low PI groups (HPIG and LPIG). 2 ´ 2 mating combinations were conducted: HPIG ♂ ´ HPIG ♀, LPIG ♂ ´ HPIG ♀, HPIG ♂ ´ LPIG ♀, LPIG ♂ ´ LPIG ♀. Four hundred G1 chickens (half in sex) equally from 4 mating groups were selected to IBV challenge at 1 d, 360 of which were artificially inoculated with allantoic fluid containing IBV M41 virus, while 40 as a control. Chickens were observed for 14 d and deaths were recorded. Paraffin sections were made and stained by hematoxylin-eosin (H. E.). Antibody titers of the survival chickens at 15 d were measured by the red cell agglutination inhibition test (HI). Twelve G1 chickens at 20 w equally from high and low PI groups were selected. Monocytes- macrophages were isolated and cultured, then challenged with Poly I:C. Expression of mRNA of cytokines and major histocompatibility complex (MHC) were tested by quantitative real-time PCR. 【Result】 Phagocytic ability of heterologous erythrocytes were different significantly in G0. G1 chickens were incubated according to mating groups according to PI of G0. Results of challenge in G1 showed that the mortality rate (33.3±0.05)% of progeny from HPIG ♂ ´ HPIG ♀ were significantly lower than that of progeny from LPIG ♂ ´ LPIG ♀(55.6±0.05)%. Mortality rates of progeny from LPIG ♂ ´ HPIG ♀ and HPIG ♂´ LPIG ♀were (43.3±0.05)% and (47.8±0.05)% respectively. Effect of hens on the offspring was greater than cocks. The mortality rate of progeny from HPIG ♀ was (38.3±0.04)%, which was significantly lower than that of progeny from LPIG ♀(51.7±0.04)%. Chickens showed clinical symptoms of cough, shortness of breath, loss of appetite and depression after challenged with IBV M41 for 3 d. Typical damages on the trachea and kidney of dead and sick chickens could be seen through H. E. staining. Epithelial cells appeared necrosis and empty bubble degeneration in tracheal, respectively. The control group showed no clinical symptoms and pathological changes. The antibody titers of 198 surviving challenged individuals showed that antibody titer of progeny from HPIG ♀ (8.45±0.07) was significantly higher than that of progeny from LPIG ♀(8.10±0.08). Expressions of IFN γ and IL-1β of high phagocytic chickens (progeny of HPIG ♀) was 5.14 times (P<0.05) and 2.41 times (P<0.05) higher than the low phagocytic chickens (progeny of LPIG ♀). Expression of MHC Ⅰ of high phagocytic chickens was significantly higher than that of low phagocytic chickens, while expression of MHCⅡ was not significant. 【Conclusion】 The experiment was performed with four mating combinations according to phagocytic index of monocytes-macrophages in vitro. Mortality rate of progeny of HPIG ♀ was significantly lower than that of progeny of LPIG ♀ after challenge, while the antibody titer and the expression of cytokines (IFN γ and IL-1β) and MHC Ⅰ were significantly higher. The results showed that the progeny of HPIG ♀ was more resistant to IBV than progeny of LPIG ♀. Therefore, the phagocytic ability of monocytes-macrophages could be an indicator for breeding IBV resistant lines.
    Effects of Lighting Regimes on Production Performance, Carcass Performance and Anti-Oxidant Capacity of the Blood in Peking Ducks
    XIN Hai-rui, PAN Xiao-hua, BI Ye, XIONG Ben-hai, JIANG Lin-shu
    Scientia Agricultura Sinica. 2016, 49(23):  4638-4645.  doi:10.3864/j.issn.0578-1752.2016.23.016
    Abstract ( 393 )   HTML ( 3 )   PDF (390KB) ( 410 )   Save
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    【Objective】As a typical meat duck variety in China, the amount of Peking Duck is increasing year by year, but there is no standardized lighting parameters for Peking Ducks at present. This trial was conducted to investigate the effects of lighting regimes on production performance, carcass performance and anti-oxidant capacity of the blood in Peking Ducks, aiming to provide a reliable theoretical basis for practical production of Peking Ducks. 【Method】Through single factor randomized block design in this experiment, three hundred and twenty 1-day-old Peking Ducks were randomly divided into 5 treatments, each treatment included 4 replicate groups with 16 ducks per group (half were male). All ducks were exposed to 5 different lighting regimes treatments: intermittent lighting (INT L), increasing lighting (INC L), short-time lighting (ST L), decreasing lighting (DEC L), and continuous lighting (CON L). The lighting intensity was 5 lx, and incandescent lamp was used as lighting source conformably. Feed consumption and body weight gain of 1-2 w, 3-5 w, 6 w and 1-6 w were recorded, and the average feed intake (AFI), average body weight gain (ABWG), feed-weight ratio (F/B) of different stages were calculated. At the end of 6 weeks, six Peking Ducks (half were male) were selected randomly from replicate groups, and those ducks were weighed and slaughtered to determine carcass traits, including dressing percentage, eviscerated percentage, breast muscle percentage, thigh meat percentage and abdominal fat percentage; Blood samples were collected from the bottom wing of four Peking Ducks (half were male) in every replicate groups. Blood samples were centrifuged promptly to determine the levels of Mel, MDA, GSH-Px and the activity of T-SOD. 【Result】The results were shown as follows: For the whole growth stage (1-6 w), lighting regimes had no significant effect on AFI, ABWG and F/B (P>0.05). During brooding period (1-2 w), the AFI and ABWG of ST L groups were significantly lower than INT L, DEC L and CON L groups. The FI of INC L group was obviously lower than INT L and DEC L groups, while the F/B was significantly higher than other groups. Lighting regimes had no significant effects on Peking Ducks’ performance during the growth (3-5 w) and fattening period (1-2 w) (P>0.05). Carcass traits (P>0.05), including dressing percentage, breast muscle percentage, thigh meat percentage and abdominal fat percentage weren’t affected by lighting regimes. However, the eviscerated percentage of CON L groups was significantly lower than other groups. There were no obvious difference in the levels of Mel, MDA and GSH-Px. The activity of T-SOD of ST L group was much lower than INT L, DEC L and CON L groups, and INC L group had lower T-SOD activity than DEC L group. 【Conclusion】 Although short-time lighting has no beneficial effect on AFI and ABWG in brooding period, it doesn’t affect the final slaughter weight of Peking ducks. The Increasing Lighting can improve the AFI in brooding period. Short-time lighting and increasing lighting decreased the activity of T-SOD and were adverse to improve anti-oxidant capacity of Peking Ducks. Besides, 24 h continuous lighting decreased the eviscerated percentage and meat production rate.
    Identification and Biosynthetic Pathway of a Hunger Pheromone in Honeybee Queen and Drone Larvae
    HE Xu-jiang, JIANG Wu-jun, YAN Wei-yu, ZENG Zhi-jiang
    Scientia Agricultura Sinica. 2016, 49(23):  4646-4655.  doi:10.3864/j.issn.0578-1752.2016.23.017
    Abstract ( 417 )   HTML ( 2 )   PDF (1403KB) ( 654 )   Save
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    【Objective】 The objective of this study is to identify the hunger pheromone of honeybee queen and drone larvae and its biosynthetic pathway, which would enormously contribute to understanding of the mechanism of communication between adults and larvae in honey bees Apis mellifera. 【Method】Two- and four-day-old queen and drone larvae and their food were collected and divided into three groups: fed larvae, starving larvae and food. For the fed larvae they were lain on their relative foods prepared in advance, and for the starving larvae their foods were totally deprived. All samples were immediately put into 20 mL sealed glass bottles and were kept in an incubator under 35 for 45 min. Afterward, a needle trap system was employed to extract 10 mL gas from those bottles and the volatile chemicals were enriched in needles. The needles were injected into a gas chromatography-mass spectrometry system and the chemicals were dissociated by a high temperature of 250 for identifying the hunger pheromone of honeybee queen and drone larvae. RNA-Seq was used for identifying the biosynthetic pathway of their hunger pheromone and the expression of related genes. 【Result】Nine and ten chemicals were identified in drone larvae and queen larvae, respectively, in which queen larvae had one more chemical (2-heptanone) that was the highest royal jelly. E-β-ocimene was identified as the hunger-signal pheromone of queen and drone larvae, since their starving larvae had significantly more E-β-ocimene than related fed larvae. The E-β-ocimene released from queen and drone starving larvae was not significantly different at 2-day-old, but queen larvae released significantly less E-β-ocimene than drone larvae at 4-day-old. Other eight chemicals were myristic acid, palmitic acid, methyl palmitic ester, stearic acid, palmitoleic acid, pentadecanoic acid, acetic acid and ethyl acetate, but did not show a clear pattern that significantly more amount of these chemicals were detected in starving larvae released than fed larvae and food groups. Acetic acid and ethyl acetate were detected higher in food and fed larvae groups compared to starving larvae groups, indicating that these two chemicals may be from their food rather themselves. RNA-Seq analysis showed that there was a de novo E-β-ocimene biosynthetic pathway in queen and drone larvae via a mevalonate pathway. Nine genes such as Geranylgeranyl pyrophosphate synthase-like and Farnesyl pyrophosphate synthase were involved in this biosynthetic pathway, but all these genes were not significantly differentially expressed between starving larvae and their relative fed larvae. 【Conclusion】Honeybee queen and drone larvae both use E-β-ocimene as their hunger pheromone for food begging, and they have a de novo E-β-ocimene biosynthetic pathway. Further, nurse honeybees specifically use 2-heptanone to mark queen larvae for guiding other nurses. This study will enrich our understanding of the biological characteristics of honeybees.
    RESEARCH NOTES
    Bioinformatics and Expression Analysis of the Cellulose Synthase Supergene Family in Flax
    YUAN Hong-mei, GUO Wen-dong, ZHAO Li-juan, YU Ying, WU Jian-zhong, ZHANG Li-guo, CHENG Li-li, ZHAO Dong-sheng, WU Guang-wen, GUAN Feng-zhi
    Scientia Agricultura Sinica. 2016, 49(23):  4656-4668.  doi:10.3864/j.issn.0578-1752.2016.23.018
    Abstract ( 445 )   HTML ( 5 )   PDF (4513KB) ( 491 )   Save
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    【Objective】 The CesA/Csls genes were identified in flax. Then the phylogeny, gene structure and tissue expression pattern were analyzed in order to provide a theoretical basis for studying the mechanism of flax fiber development.【Method】Based on flax genome database and bioinformatics method, CesA/Csls genes were identified and the physico-chemical characteristics were analyzed. The phylogenetic tree was constructed by MEGA 5.0 software. The gene structure and conservative motifs were analyzed by the bioinformatics softwares GSDS and MEME. Finally, the expression of CesA/Csl genes was analyzed by using the RNA-seq data. 【Result】 A total of 45 CesA/Csl genes were systematically identified in flax. The genes appeared to be dispersed within the chromosome and were not clustered. The CesA/Csl proteins were mainly located on the plasma membrane. The number of amino acid of the proteins ranged from 409 to 1 167. The isoelectric point distributed from 5.43 to 9.08. All of the 45 CesA/Csl proteins possessed the transmembrane domains, the number of which was from 2 to 8. The genes were classified into 2 classes (CesA and Csl) and seven groups (CesA, CslA, CslB, CslC, CslD, CslE, CslG) according to the phylogetic relationship. Gene structure prediction indicated that CesA/Csls genes ranged from 2.1 to 6.8 kb in size and most of them consist of 2 to 14 exons.The gene structure was conserved within a group. Obvious differences were observed in motif composition in genes from different groups. Motif 1 to motif 4, motif 12 were observed in most of CesA, CslB, CslD, CslE, and CslG group proteins,and motif 18, and motif 20 were observed in most of CslA, CslC group proteins. Motif 13, motif 14, motif 15, and motif 19 as the spectific motifs were observed in different groups. Futhermore, some CesA/Csl genes could be upregulated or downregulated by BR, Brz and NaCl stress. The results of digital gene expression profile showed that CesA/Csl were expressed differently at different development stages. It indicated that CesA/Csls had various functions and played different roles in the process of plant development. 【Conclusion】A total of 45 CesA/Csl genes in flax were systematically identified by genome-wide screening. The genes were classified into 2 classes and seven groups. The genes appeared to be dispersed within the scaffolds. The analysis of gene structure and conservative motifs indicated that there were obvious differences between groups, but the genes within a group varied slightly. RNA-seq expression profiling of the CesA/Csl genes revealed unique, homoeolog-specific expression patterns at different development stages. Some of them could be induced by BR, Brz or NaCl stress.
    Functional Response and Numerical Response of Two Geographic Populations of Trichogramma dendrolimi to the Eggs of Rice Moth, Corcyra cephalonica
    DU Wen-mei, ZANG Lian-sheng, ZHANG Jun-jie, QI Ying-hui, RUAN Chang-chun
    Scientia Agricultura Sinica. 2016, 49(23):  4669-4676.  doi:10.3864/j.issn.0578-1752.2016.23.019
    Abstract ( 359 )   HTML ( 1 )   PDF (394KB) ( 355 )   Save
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    【Objective】 Trichogramma dendrolimi is a widely used parasitoid in biological control. There are some differences in different geographic populations of T. dendrolimi and its biological characteristics. This study aimed at understanding the parasitic functional response and numerical response of two typical geographic populations to eggs of the rice moth, Corcyra cephalonica so that provided a reference for the application of different geographical populations of the parasitic wasp species.【Method】 C. cephalonica eggs were used as the host, and the functional response and numerical response of two geographic populations of T. dendrolimi from Heilongjiang and Jiangsu parasitizing were investigated. The species flocks of trend index (I) and index of population control (IIPC) of the two different geographical populations were compared and analyzed.【Result】 The parasitic functional responses of Heilongjiang population (TD-HLJ) and Jiangsu population (TD-JS) of T. dendrolimi fitted in with HollingⅡ equation. The daily maximum parasitic number of TD-HLJ was 186.78 eggs, more 75.68 eggs than that of TD-JS. The density of 1 grain of C. cephalonica egg was the critical value of TD-HLJ and TD-JS populations of eclosion. The density of C. cephalonica eggs had significant impact on parasitism, development and reproduction of both populations. 5 and 10 eggs of C. cephalonica were the critical values of TD-HLJ and TD-JS population reproduction. The population trend index (I) of both populations increased with the increase of host egg density. Generally, both of populations had the values of I under the control density with 68.70 for TD-HLJ and 49.20 for TD-JS, which indicated that the number of offsprings of TD-HLJ was 19.5 times of that of TD-JS. Similarly, interference of index of population control (IIPC) of both populations increased with the increase of host egg density, indicating that the biological control effectiveness of T. dendrolimi against hosts decreased with egg density increasing.【Conclusion】 Based on the comprehensive analysis on the results of functional response and numerical response, TD-HLJ exhibits better biological control potential than TD-JS in the insect pest management.