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Table of Content

    15 July 2013, Volume 46 Issue 14
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Research Progresses in Ac/Ds Transposon Activation Tagging System
    WANG Wen-Jing, MA Hao-Ran, LI Jia-Na, ZHANG Hong-Bo
    Scientia Agricultura Sinica. 2013, 46(14):  2845-2855.  doi:10.3864/j.issn.0578-1752.2013.14.001
    Abstract ( 700 )   PDF (564KB) ( 1514 )   Save
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    Along with the development of plant functional genomics, creation of an informative mutant population is becoming more and more important. The Ac/Ds transposon system is an effective tool for mutant creation, especially for plant species with difficulties in transformation. And, the development of Ac/Ds transposon activation tagging system makes it feasible to create dominant mutations. In this paper, the progresses in the development of Ac/Ds transposon activation tagging system with versatile activation tagging were introduced and the future perspectives of this system in creation of plant mutant populations were discussed.
    Cloning and Expression Analysis of Sugarcane Phenylalanin Ammonia-lyase (PAL) Gene
    SONG Xiu-Peng-1, HUANG Xing-2, MO Feng-Lian-1, TIAN Dan-Dan-3, YANG Li-Tao-12, LI Yang-Rui-12, CHEN Bao-Shan-1
    Scientia Agricultura Sinica. 2013, 46(14):  2856-2868.  doi:10.3864/j.issn.0578-1752.2013.14.002
    Abstract ( 679 )   PDF (1371KB) ( 1373 )   Save
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    【Objective】The aim of this study was to clone full-length cDNA of sugarcane phenylalanin ammonia-lyase (PAL) gene (ScPAL), a key enzyme gene related to phenylpropanoid metabolism in sugarcane, investigate its sequence characteristics, analyze its expressions in different organs and under five different stress conditions, thus providing a theoretical support for using this gene in sugarcane stress tolerance breeding. 【Method】 The differentially expressed protein was identified by MALDI TOF/TOF and analyzed using Applied Biosystems GPS Explorer software with Mascot analysis against the NCBI and Uniprot database using a combined peptide mass fingerprint and MS/MS search, then the full sequence of ScPAL was cloned from sugarcane variety ROC22 using RT-PCR and RACE techniques. The bioinformatics method was used to analyze the putative amino acid sequence, and real-time PCR method was used to analyze the expression of ScPAL in different tissues and under different stresses. 【Result】 The full-length cDNA of ScPAL (GenBank accession number: KC172559) in sugarcane was cloned. The sequence consists of 2590 bp with an intact open reading frame of 2115 bp, encoding a polypeptide of 704 amino acids. Sequence analysis showed that it contains the typical PAL enzyme active site sequence (GTITASGDLVPLSYIA). Homology analysis showed that the deduced ScPAL protein was highly homologous to other PAL proteins from different species. Phylogenetic tree analysis indicated that ScPAL was very closely related to PAL of sorghum. Real-time PCR results showed that the ScPAL expressed in root, stalk and leaf, respectively, and its expression was different among three organs. The mRNA of ScPAL in root was the highest among three organs and was about sixty-six times higher than that in leaf. Furthermore, ScPAL transcription level was induced under the treatment of low temperature, PEG, NaCl and H2O2 stresses, but the expression patterns were different. 【Conclusion】 The gene ScPAL which was firstly cloned and characterized from sugarcane (ROC22) is a member of PAL family typically. The results of study indicated that it participated in sugarcane resistance to smut, also played a role in the sugarcane resistance to chilling, drought and salt stress processes.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Effects of High Temperature at Different Times During the Heading and Filling Periods on Rice Quality
    ZHANG Gui-Lian, ZHANG Shun-Tang, WANG Li, XIAO Ying-Hui, TANG Wen-Bang, CHEN Guang-Hui, CHEN Li-Yun
    Scientia Agricultura Sinica. 2013, 46(14):  2869-2879.  doi:10.3864/j.issn.0578-1752.2013.14.003
    Abstract ( 528 )   PDF (1103KB) ( 985 )   Save
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    【Objective】The effects of high temperature treatment at different times during heading and filling periods on rice quality were studied to ascertain the mechanism of the effect of high temperature on rice quality and its time effect. 【Method】The experiment was conducted with two rice lines, the heat-tolerant line 996 and heat-sensitive line 4628, in controlled environment chambers to investigate the effect of high temperature stress at different times during heading and filling stages on rice appearance quality, milling quality, amylose content, gel consistency, protein content, RVA characteristcs, Mg/K and starch granule structure of grain.【Result】The ratio of grain length to width, the brown rice rate, milled rice rate, head rice rate and gel consistency of both lines decreased under high temperature stress during heading and filling stages, while chalkness rate, chalkness, amylose content, protein content and gelatinization temperature increased. Under high temperature treatments, gelatinization temperature, final viscosity, set back and peak time increased, break dowm decreased, Mg content and K content increased, Mg/K decreased. The observation of starch structure with scanning electron microscope showed that the starch granules arranged loosely, most of the starch granules existed in the form of single starch granule, refractive index decreased, endosperm transparency decreased, and chalkiness was formed. Under the same treatment, the extent of rice quality of heat-tolerant line 996 affected by high temperature was lower than that of heat-sensitive line 4628. Rice qualities had different responses to different high temperature treatments, rice qualities were affected by high temperature treatment 8-14 d after heading mostly, moderatedly by high temperature treatment 15-21 d and 1-7 d after heading, while least by high temperature treatment 22-28 d after heading. 【Conclusion】Eight to twenty-one days after heading is the vital stage for whole rice quality in response to higher temperature, which affects rice qualities decisively; while the effect of temperature at the later stage(beyond 22 days from heading) is slight.
    Study on Maize Yield Estimation and Accuracy Assessment Based on PyWOFOST Crop Model in Northeast China
    CHEN Si-Ning-123, ZHAO Yan-Xia-2, SHEN Shuang-He-3, LI Zhen-Fa-1
    Scientia Agricultura Sinica. 2013, 46(14):  2880-2893.  doi:10.3864/j.issn.0578-1752.2013.14.004
    Abstract ( 723 )   PDF (1036KB) ( 812 )   Save
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    【Objective】 The objective of this study is to construct a reasonable crop yield estimation scheme and improve the accuracy of crop yield estimation. 【Method】Based on the PyWOFOST crop model developed according to Ensemble Kalman Filter(EnKF) which combined remote sensing information with crop model, the PyWOFOST model was modified and improved to take LAI as the joint point of the crop model(WOFOST) and remote sensing information and be applicable to the maize-growing area in Northeast China. MODIS LAI data were used as external assimilation data to simulate maize LAI, yield and development stage with the PyWOFOST model at agro-meteorological stations. The impact of uncertainty(i.e. random error) of remote sensing observations(MODIS LAI) and model parameter(thermal time from emergence to anthesis, TSUM1) on the assimilation simulation results was analyzed deeply. Finally, the PyWOFOST model was used to estimate maize yield on a regional scale. 【Result】The result shows that, the modeled maize yield after assimilating MODIS LAI was closer to the observed values by comparing with the results modeled by WOFOST, the errors of maize production before and after assimilation at different uncertainty levels of TSUM1(0, 10, 20, 30℃) of 20 agro-meteorological stations without the impact of meteorological disasters were14.04%, 12.71%, 11.91%, 10.44% and 10.48%, respectively. LAI modeled by PyWOFOST after assimilation which more in line with the change trend of maize LAI was generally closer to the observed LAI than LAI modeled by WOFOST. The mean error of development stage between modeled value of WOFOST and observed value was 3.4 d; the mean errors of development stage between modeled value of PyWOFOST at different uncertainty levels of TSUM1(0, 10, 20, 30℃) and observed value were 3.5, 4.3, 5.0 and 5.5 d, respectively. The model results on the regional scale showed that, the errors of maize yield with assimilation were less than 15% in 58.82% of the study area; the coefficient of determination between modeled yield with assimilation and statistical yield was 0.806. 【Conclusion】Overall, the simulation results on the stations and regional scales both revealed the advantages of the crop yield estimation based on EnKF assimilated remote sensing information into crop models.
    PLANT PROTECTION
    Mechanisms of Rice Resistance (Susceptibility) Manipulated by Diverse TALEs of Xanthomonas oryzae pv. oryzae and pv. oryzicola and Potential Utilization in Rice Breeding
    LI Zheng, XIONG Li, JI Zhi-Yuan, ZOU Li-Fang, ZOU Hua-Song, CHEN Gong-You
    Scientia Agricultura Sinica. 2013, 46(14):  2894-2901.  doi:10.3864/j.issn.0578-1752.2013.14.005
    Abstract ( 518 )   PDF (786KB) ( 1081 )   Save
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    Breeding and planting resistant rice varieties is one of the most effective and economic ways to control bacterial leaf blight and bacterial leaf streak diseases caused by Xanthomonas oryzae pv. oryzae (Xoo) and pv. oryzicola (Xoc), respectively. Recent land-marked advances in understanding the mechanism of rice resistance (R) or susceptibility(S) manipulated by transcriptional activator-like effector (TALEs) of Xoo and Xoc shall definitely alter present strategies in rice breeding for disease resistance. Hereby, the authors reviewed the corresponding relationships between known TALEs of a Xoo model strain PXO99A and known and unknown R or S genes in rice, forwarded the prospects regarding to co-evolution of new tale genes in the pathogens and to mine R or S genes in rice, and introduced a TALEN technology, developed based on DNA-binding specificity of TALEs, that has been used to genetically transfer a susceptible rice line into a resistant one. The TALEN technology is tremendously a powerful tool that will realize such a goal in altering susceptible rice varieties (materials) with high yield and high quality into resistant ones with high yield and high quality.
    Advances in Research of Genetic Diversity and Pathogenome of Ralstonia solanacearum Species Complex
    XU Jin, FENG Jie
    Scientia Agricultura Sinica. 2013, 46(14):  2902-2909.  doi:10.3864/j.issn.0578-1752.2013.14.006
    Abstract ( 590 )   PDF (616KB) ( 1252 )   Save
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    Bacterial wilt, caused by Ralstonia solanacearum, is one of the most devastating plant diseases worldwide. As a species complex, R. solanacearum strains are highly variable and adaptable as attested by worldwide distribution and a wide and expanding host range. A new phylotype classification scheme was proposed by Fegan and Prior to distinguish the genetic diversity within the R. solanacearum species complex. High genome complexity and plasticity of R. solanacearum is reflected by its great adaptation potential to different hosts and environments. This review includes recent advances in research of genetic diversity and pathogenome of R. solanacearm species complex.
    DSF Signal-Dependent Quorum Sensing in Plant Pathogenic Bacteria Xanthomonas
    ZHOU Lian, WANG Xing-Yu, HE Ya-Wen
    Scientia Agricultura Sinica. 2013, 46(14):  2910-2922.  doi:10.3864/j.issn.0578-1752.2013.14.007
    Abstract ( 559 )   PDF (823KB) ( 1446 )   Save
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    It is now clear that quorum sensing (QS) is one of the most important cell-cell communications in bacteria and this community-wide genetic regulatory mechanism is involved in the regulation of many important biological activities. Since the 1980s, several types of QS signals have been identified. The diffusible signal factor (DSF),which was originally identified in Xanthomonas campestris pv. campestris (Xcc), represents another interesting type of QS signals found in Gram-negative bacteria. The rapid progress over the last 10 years has identified the chemical structures of the QS signal DSF, established the DSF regulon, and unveiled the general signaling pathways and mechanisms. RpfF is a key enzyme for DSF biosynthesis, which is modulated by a novel autoinduction mechanism involving protein-protein interaction between the DSF synthase RpfF and the sensor RpfC. DSF signal is sensed by RpfC/RpfG two component system and the signal sensing is coupled to intracellular regulatory networks through a second messenger cyclic di-GMP and a global regulator Clp. Genomic and genetic analyses show that the DSF signal regulates diverse biological functions including virulence, biofilm dispersal, and ecological competence. Evidence is emerging that the DSF QS system is conserved in a range of plant and human bacterial pathogens.
    Advances in Research of Rice Bacterial Foot Rot
    LIU Qiong-Guang, ZHANG Qing, WEI Chu-Dan
    Scientia Agricultura Sinica. 2013, 46(14):  2923-2931.  doi:10.3864/j.issn.0578-1752.2013.14.008
    Abstract ( 622 )   PDF (561KB) ( 1277 )   Save
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    Rice bacterial foot rot caused by Dickeya zeae is an important bacterial disease in China and Southeast Asian countries. It occurs severely in different years in local areas. This paper reviews the research progress about the disease occurrence situation in China, the classification of D. zeae, and its infection characteristics, host range, pathogenic factors, virulence and its regulation. In addition, the investigations of some problems involving in the disease occurrence and control were presented.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Simulation of Long-Term and Regional Environmental Effects of Different N Applications in the Winter Wheat/Summer Maize System
    WANG Li-Gang, LI Hu, YANG Li, DI Zhen, QIU Jian-Jun
    Scientia Agricultura Sinica. 2013, 46(14):  2932-2941.  doi:10.3864/j.issn.0578-1752.2013.14.009
    Abstract ( 488 )   PDF (668KB) ( 1135 )   Save
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    【Objective】 As the overuse of N application in the winter wheat/summer maize rotation system and the enhanced environmental effects, this research aims at the long-term and regional environmental effects of different N applications, and to definite the regional environmental effects and the potential of N reduction in the winter wheat/summer maize system.【Method】The simulation of different N application in winter wheat/summer maize was conducted for 15 years by using the research method which combined the location tests and the biogeochemical model together. 【Result】 The result indicated that crop yield increased by increasing N application. The increase of crop yield was limited when the N application rate ranged from 240 kg•hm-2 to 300 kg•hm-2, and the N leaching enhanced, the residual inorganic N in the soil increased. The loss of total N reduced under the two regulations, 30% i.e. the regional N application rate reduced by 30% and the optimized N application, which lead to 67.23% and 79.93% reduction of N leaching, respectively, thus decreasing the N environmental risk.【Conclusion】The N environmental effects of the winter wheat/summer maize rotation system increased by increasing N application in the area around Bohai Bay. The planting pattern of winter wheat/summer maize has the potential of N reduction, and reduce 30% of regional N application has the obvious reduction of environmental effects. The middle of Hebei Province and the east and southwest of Shandong Province have the most obvious potential of saving N.
    Effects and Competitive Nodulation Ability of Inoculation with Rhizobium leguminosarum Strains on a Semi-Leafless Pea
    GUO Li-Mei-12, YANG Yong-2, ZHANG Shi-Chen-2, HU Yue-Gao-2, SUI Xin-Hua-3, ZENG Zhao-Hai-2
    Scientia Agricultura Sinica. 2013, 46(14):  2942-2952.  doi:10.3864/j.issn.0578-1752.2013.14.010
    Abstract ( 472 )   PDF (728KB) ( 541 )   Save
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    【Objective】The objective of this study is to determine the effects of inoculating pea with different Rhizobium leguminosarum strains on pea’s growth and development, yield, grain quality, the competitive nodulation ability of the inoculated strains, and rhizobium strains best matching with the pea variety. 【Method】 A pot experiment was conducted by inoculating a Canadian semi-leafless pea variety 'MISER' with 9 R. leguminosarum strains (CCBAU 33109, CCBAU 43228, CCBAU 43227, CCBAU 43232, ACCC 16058, CCBAU 41242, CCBAU 43242 and 2 commercial rhizobia F98-1, F98-4) in low nitrogen soil from a cotton field in Huarong County, Hunan Province. Growing stages of pea were investigated. Pea’s main growth characteristics, nodulation and nitrogen fixation performance were measured at full-flowering stage, and meanwhile, the nodule occupancy of each inoculated strain was assayed using BOX-PCR fingerprinting technology. Pea’s main growth characteristics, yield, yield components, crude protein and crude fiber content of dry grains were determined at maturing stage. 【Result】 Compared with not-inoculated treatment (CK), the podding and pod ripening periods of pea inoculated with CCBAU 33109 were significantly increased by 4.1 and 7 d (P<0.05), respectively. At pod ripening period, the numbers of main stem nodes of pea inoculated with ACCC 16058 were significantly increased by 2.8, 3.8 and 2.9, compared with CK and treatments inoculated with F98-1 and F98-4, respectively. The height of pea inoculated with ACCC 16058 increased significantly by 15.67% compared with F98-4. Compared with CK and the treatment inoculated with F98-1, inoculating with ACCC 16058 significantly increased MISER’s crude protein content by 9.88% and 10.63%, respectively. The grain fresh weight per plant of peas inoculated with CCBAU 41242 was also significantly 40.46% heavier than that of peas inoculated with F98-1. The nodule occupancy of CCBAU 43228 was 57.50%, higher than that of any other strain. 【Conclusion】Based on the analysis above, rhizobia CCBAU 33109, ACCC 16058, CCBAU 41242 and CCBAU 43228 matched better with the pea variety ‘MISER’ than other strains.
    HORTICULTURE
    Cloning and Expression Analysis of A Male Sterility-Related Gene VvMS2 from Grapes
    ZHENG Huan-1, ZHANG Ji-Yu-12, WANG Xin-Wei-1, ZHANG Zhen-1, JI Chen-Fei-1, TAO Jian-Min-1
    Scientia Agricultura Sinica. 2013, 46(14):  2953-2962.  doi:10.3864/j.issn.0578-1752.2013.14.011
    Abstract ( 510 )   PDF (1111KB) ( 1234 )   Save
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    【Objective】 This paper aims to isolate the VvMS2 gene from the Vitis vinifera L. cv. Wink and Zhongshanhong, in order to further study the molecular mechanism of grape male sterility and grape molecular breeding. 【Method】 Specific primers were designed based on the grape sequence (CBI29968) in NCBI which is highest homology with Arabidopsis male sterility gene MS2, and the cDNA and genomic DNA of VvMS2 were cloned by PCR. VvMS2 gene structure characteristics were analyzed using the bioinformatics software. Quantitative real-time PCR (qRT-PCR) was performed to determine the expression pattern of VvMS2 in different tissues of ‘Wink’ and at different development stages in bud between ‘Zhongshanhong’ and ‘Wink’.【Result】The full length ORF cDNA sequence of VvMS2 with 1 755 bp were cloned. Bioinformatics analysis showed that this gene encodes 584 amino acids, containing NAD-binding-4 structure domains and sterile structure domains. This gene contains 9 exons and 8 introns. The protein molecular weight is 64.74 kD, isoelectric point is 8.84. Compared with other sequences of MS2 in GenBank, the deduced amino acid sequences of VvMS2 shared 40%-71% in homology. VvMS2 only specially expressed in floral buds and at the stages of tetrad to dinucleate. The expression level in button at uninucleate stage in ‘Wink’ is remarkably higher than that in ‘Zhongshanhong’.【Conclusion】The abnormal gene expression is related with male sterility in ‘Zhongshanhong’.
    Effect of Grafting on Secondary Metabolism and Its Relationship with Bacterial Wilt Resistance in Pepper
    LIU Ye-Xia-1, FU Ling-1, AI Xi-Zhen-1, WANG Hong-Tao-2, JI De-Gang-2
    Scientia Agricultura Sinica. 2013, 46(14):  2963-2969.  doi:10.3864/j.issn.0578-1752.2013.14.012
    Abstract ( 485 )   PDF (563KB) ( 538 )   Save
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    【Objective】 The purpose of this study is to elucidate the phenolic acids metabolism in grafted pepper and its function in improvement of bacterial wilt resistance. 【Method】 Changes of the electrolyte leakage (EL), hydrongen peroxide (H2O2) content, phenolic acids and activities of the related enzymes in grafted (‘Xinfeng 2’ scion grafted onto ‘Weishi’) and own-root plants (‘Xinfeng 2’, control) of pepper were investigated by manual inoculation. 【Result】The results showed that the EL in leaves of grafted and own-root peppers all increased gradually after inoculation, whereas the increasing range in grafted plants was less than that in control plants. Grafted pepper showed an increase in H2O2 contents in leaves and roots at early stage, but a decrease after 6-8 days of inoculation. Compared with the control plants, grafted plants showed higher contents of vanillin, salicylic acid and lignin, and activities of phenylalanine ammonia lyase (PAL) and polyphenoloxidase (PPO). 【Conclusion】The increased phenolic acids synthesis and secretion in grafted plants is one of the important reasons for enhancement in disease resistance. The salicylic acid and vanillin played the major role in improving disease resistance of grafted pepper plants.
    Research on Soil and Leaf Nutrient Diagnosis of ‘Red Fuji’ Apple Orchard in Two-Dominant Producing Areas
    WANG Fu-Lin, MEN Yong-Ge, GE Shun-Feng, CHEN Ru, DING Ning, PENG Fu-Tian, WEI Shao-Chong, JIANG Yuan-Mao
    Scientia Agricultura Sinica. 2013, 46(14):  2970-2978.  doi:10.3864/j.issn.0578-1752.2013.14.013
    Abstract ( 625 )   PDF (811KB) ( 689 )   Save
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    【Objective】Soil available nutrients and leaf mineral elements and nutritional status in ‘Red Fuji’ apple orchards in two dominant producing areas of Circum-Bohai and Loess Plateau in China were determined in order to conduct the analysis and diagnosis for ‘Red Fuji’ apple orchard nutritional status for guiding scientific fertilization.【Method】Based on soil available nutrient classification standards and DRIS nutrient diagnosis, soil and leaf samples from high and low yield orchards of 15-20-year old trees of two apple producting areas were analyzed. Differences among treatments were separated by the least significant difference (LSD) test, and One-Way Anova and difference analysis were conducted by DPS V7.05.【Result】The organic matter, alkaline hydrolysis N, available P, available K of soil available nutrient content average values were 10.9 g•kg-1, 73.21 mg•kg-1, 70.22 mg•kg-1, and 169.2 mg•kg-1 in Circum-Bohai, respectively, while the soil available nutrient content average values were 11.7g•kg-1, 56.46 mg•kg-1, 14.91 mg.kg-1, and 135.78 mg•kg-1 in Loess Plateau, respectively. The alkaline hydrolysis N, available P, available K in Circum-Bohai were higher than those in Loess Plateau, except organic matter. The N, P, K, Ca, Mg, Fe, Mn, Cu, Zn, B, Mo content of average values were 2.73%, 0.18%, 1.15%, 9.63 g•kg-1, 2.86 g•kg-1, 87.97 mg•kg-1, 124.02 mg•kg-1, 13.36 mg•kg-1, 15.64 mg•kg-1, 35.09 mg•kg-1, and 0.51 mg•kg-1 in Circum-Bohai, respectively. The leaf average values were 2.72%, 0.18%, 1.17%, 10.23 g•kg-1, 2.99 g•kg-1, 88.69 mg•kg-1, 116.39 mg•kg-1, 10.85 mg•kg-1, 15.94 mg•kg-1, 39.55 mg•kg-1, and 0.51 mg•kg-1 in Loess Plateau, respectively. The DRIS diagnostic results in two-dominant producing areas indicated that the most deficient leaves elements were Ca and K in low-yielding orchard of Circum-Bohai, followed by Fe, N, and Zn, and the most wealthy elements were Cu and Mo. The most lacking elements were P and K in Loess Plateau, followed by N, Zn, and Cu, and the most wealthy elements were B and Ca.【Conclusion】It was concluded that the organic contents in two regions were slightly low, the soils are deficient in N. P is rich and K is deficient in soils in Circum-Bohai region, while P is poor and K is deficient in soils in Loess Plateau. Leaf diagnosis of low-yield orchards showed that the nutrition at status of trees in Loess Plateau was better than that of the Circum-Bohai , and both areas had a great potential of production. It was suggested that fertilizer of K, N, Fe, Ca, and Zn in Circum-Bohai should be increased, and fertilizers of K, P, Zn, Cu, and Fe should be increased in Loess Plateau, meanwhile organic manure should be attached to the along with chemical fertilizer.
    Spatial and Temporal Accumulation of Flavanols, Activity and Tissue Localization of Leucoanthocyanidin Reductase Induced by Soil Drought in Developing Grape Berries
    WEN Peng-Fei, NIU Xing-Yan, XING Yan-Fu, GAO Mei-Ying, NIU Tie-Quan
    Scientia Agricultura Sinica. 2013, 46(14):  2979-2989.  doi:10.3864/j.issn.0578-1752.2013.14.014
    Abstract ( 534 )   PDF (968KB) ( 617 )   Save
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    【Objective】The effects of soil drought on the spatial and temporal accumulation of flavanols, enzyme activity and tissue localization of its biosynthetic key enzyme, leucoanthocyanidin reductase (LAR), during the grape berry development were studied.【Method】The 5-year old grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) was used as materials, and the grapevine was subjected to soil drought stress simulated by controlling irrigation through the rain-preventing shelter and root-cutting groove during grape berry development. The spatial and temporal accumulation of flavanols and the enzyme activity of LAR in different parts of the grape berry were analyzed by spectrophotometer method, and the tissue localization of LAR was detected by the immunohistochemical localization. 【Result】 Results indicated that the growth of grape berry was significantly inhibited by soil drought during its development, and an obvious accumulation of total phenol in berry was induced by soil drought, especially in the young berry period. The accumulation pattern of flavanols and total flavan-3-ols in skin and flesh during grape berry development were not changed by soil drought, whereas an obvious accumulation of flavanols and total flavan-3-ols induced by soil drought was observed, and this effect was organ- and development-dependent. The LAR enzyme activity in skin and flesh was induced by soil drought, especially in the young berry period. There were no obvious changes of tissue localization of LAR1 and LAR2 in the skin and flesh under soil drought, but an increasing signal was detected, especially in the vascular bundle.【Conclusion】All the results of this study suggested that soil drought could induce the accumulation of LAR1 and LAR2 proteins in different parts of berry at different development stages, resulted in the increase of LAR enzyme activity in corresponding part and developing stage, and induced the accumulation of flavanols and total flavan-3-ols during grape berry development.
    STORAGE·FRESH-KEEPING·PROCESSING
    Cytotoxicity and Anti-Proliferation Effect on HepG2 Cells and Cellular Antioxidant Activity(CAA) of Adlay Polyphenols
    WANG Li-Feng-1, CHEN Jing-Yi-1, XIE Hui-Hui-1, WANG Yu-Mei-1, SHI Jia-Yi-1, LIANG Zheng-Chang-2, LIU Rui-Hai-2, JU Xing-Rong-1, YUAN Jian-1
    Scientia Agricultura Sinica. 2013, 46(14):  2990-3002.  doi:10.3864/j.issn.0578-1752.2013.14.015
    Abstract ( 504 )   PDF (827KB) ( 1275 )   Save
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    【Objective】The objective of this study is to learn and analyze the antioxidant capacity of adlay extracts, especially that of the bound part , and to determine the contribution of the bound part to the total antioxidant capacity. The toxicity and anti-proliferative effect of adlay polyphenols on liver cancer cells were also studied. In the meanwhile, the functions of adlay extracts were evaluated and analyzed at cell level. 【Method】 Guizhou heigu adlay, Liaoning 5 adlay and Longyi 1 adlay were used as raw materials, the intracellular ORAC was evaluated by building cell models of free or bound polyphenols of adlay extracts. With human hepatoma cells HepG2 as the research object, CC50 for cytotoxicity and EC50 for anti-proliferative effect were determined and analyzed.【Result】The antioxidant capacity of bound polyphenol is significantly higher than that of the free part. The content of totel polyphenol and the antioxidant capacity are obviously different among different species. Liaoning 5 adlay and Longyi 1 adlay are significantly better than Guizhou Heigu adlay. The adlay extracts have an obvious effect on inhibition of proliferation of human liver cancer cells, and substantially in the experimental concentration range, the adlay sample itself has no cytotoxicity.【Conclusion】As a traditional Chinese medical and edible whole grain foods, adlay has abundant polyphenols and strong antioxidant capacity. The antioxidant value significantly improved after considering the existence of bound polyphenols. Besides, polyphenol extracts of adlay have an obvious anti-proliferative effects on tumor cells.
    Potential Sources of β-Carbolines Harman and Norharman in Braised Sauce Meat
    PAN Han, WANG Zhen-Yu, GUO Hai-Tao, NI Na, CHEN Li, ZHANG De-Quan
    Scientia Agricultura Sinica. 2013, 46(14):  3003-3009.  doi:10.3864/j.issn.0578-1752.2013.14.016
    Abstract ( 629 )   PDF (552KB) ( 670 )   Save
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    【Objective】The potential sources of co-mutagens β-carbolines 1-methyl-9H-pyrido[3,4-b]indole (harman) and 9H-pyrido[3,4-b]indole (norharman) in braised sauce meat were investigated. 【Method】 The occurrence of the bioactive β-carbolines in six braised sauce meats and five soy sauces were studied by solid-phase extraction and HPLC-DAD. Furthermore, the potential sources of β-carbolines in braised sauce mutton boiled with one of the soy sauces were determined.【Result】β-carbolines were detected in all of the six braised sauce meats and five kinds of soy sauces. The concentrations of harman in braised sauce meat were 3.71-42.32 ng•g-1, and norharman were 2.95-63.97 ng•g-1. The levels of harman and norharman in soy sauces were 111.47-301.30 ng•g-1 and 74.71-199.27 ng•g-1, respectively. Raw mutton contained undetectable amounts of harman and norharman. However, blank mutton samples, which were boiled only with water, contained harman and norharman for 3.39 and 4.05 ng•g-1. In addition, there were 17.30 ng•g-1 harman and 12.18 ng•g-1 norharman in the control samples containing soy sauce, salt and other seasonings. The proportion of harman and norharman in boiled mutton was 15% and 23%. The percentage of harman and norharman by boiling soy sauce and seasonings was 58% and 46%, respectively. Mutton reacted with soy sauce generated 27% harman and 31% norharman.【Conclusion】Both harman and norharman appeared in braised sauce meat and soy sauce, and soy sauce contained higher content of β-carbolines. Almost 50% of β-carbolines in braised sauce mutton was due to the addition of soy sauce. The pyrolysis of amino acids in mutton and the chemical reaction of soy sauce and mutton were important sources of harman and norharman.
    ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT
    Dynamics of PPARγ, C/EBPα and Myogenin Gene Promoter Methylation During Myoblasts Intracellular Lipid Accumulation
    JIANG Mei-Hua, JU Ting-Ting, LIU Yang, XU Ling, SUN Wen-Juan, ZHAO Pan-Feng, YIN Jing-Dong
    Scientia Agricultura Sinica. 2013, 46(14):  3010-3021.  doi:10.3864/j.issn.0578-1752.2013.14.017
    Abstract ( 600 )   PDF (1163KB) ( 1755 )   Save
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    【Objective】This experiment was designed to investigate whether DNA methylation of adipogenic promoters (peroxisome proliferator-activated receptor gamma (PPARγ) and CCAAT/enhancer binding protein alpha (C/EBPα)) and myogenic promoter (myogenin) was involved in C2C12 myoblasts intracellular lipid accumulation.【Method】C2C12 cells were treated, respectively, by 2% horse serum and an adipogenesis cocktail which typically contains 3-isobutyl-methylxanthine, dexamethasone and insulin (MDI). In order to observe the formation of myotubes, cells were incubated for 0 d, 1 d, 3 d, and 5 d by 2% horse serum and then were collected to perform Giemsa staining. To confirm the lipid accumulation, cells treated by adipogenesis program for 0 d, 2 d, 4 d, 6 d and 10 d were fixed to conduct Oil Red O staining. Genome DNA and total RNA were extracted from C2C12 cells during the myogenesis differentiation and intracellular lipid accumulation. Real-time quantitative reverse transcription PCR was used to detect the mRNA levels of myogenic and adipogenic related genes. The bisulfite sequencing was adopted to explore the sequential DNA methylation changes in PPARγ, C/EBPα and myogenin promoter regions, and the relationship between DNA methylation and gene expression was analyzed.【Result】After horse serum treatment, the morphology of multinucleated myotubes and up-regulation of myogenic genes were displayed in C2C12 cells, but the adipocyte-specific genes showed no significant changes. The adipogenesis cocktail induced the intracellular lipid accumulation during automatical differentiation into myotubes of C2C12 cells, and both the myogenic and adipigenic genes were up-regulated. The bisulfite sequencing showed that the methylation level of PPARγ gene in undifferentiated myoblasts was 61%, and progressively demethylated upon adipogenic differentiation with methylation levels of 49%, 39% and 42% on d 2, 4 and 6, which was accompanied by an increase of mRNA level of PPARγ. But during the horse serum treatment, there were no significant changes in PPARγ methylation levels (56% and 48% on d 3 and 5) and mRNA levels. In the case of myogenin, it showed decreased methylation levels after horse serum inducement with methylation levels of 49%, 42%, 35% and 34% on d 0, 1, 3 and 5, respectively, while the expression of myogenin increased rapidly. During the adipogenic inducement, myogenin gene also demethylated (49%, 37%, 41% and 38% on d 0, 2, 4 and 6), but the degree was weaker than that in horse serum treatment, which agreed with the reduced up-regulation of myogenin mRNA level. C/EBPα showed hypomethylated status with only 1.6% in undifferentiated C2C12 cells and maintained the hypomethylated status regardless the myogenesis or lipid accumulation, so the methylation of C/EBPα promoter seemed unrelated to its transcription.【Conclusion】The dynamics of DNA methylation of PPARγ and myogenin promoter are closely correlated with their gene expression, which indicates that DNA methylation may play a role in regulating the myogenesis and lipid accumulation in C2C12 cells.
    Association of Single Nucleotide Polymorphism of H-FABP and HSL Genes with Growth Traits in Maiwa Yak
    CHAI Zhi-Xin-1, WANG Yong-1, LUO Xiao-Lin-2, WU Zhi-Juan-1, LIU Zhong-Na-1, ZHONG Jin-Cheng-1
    Scientia Agricultura Sinica. 2013, 46(14):  3022-3031.  doi:10.3864/j.issn.0578-1752.2013.14.018
    Abstract ( 476 )   PDF (745KB) ( 785 )   Save
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    【Objective】Correlations between SNP of Maiwa yak heart fatty acid-binding protein (H-FABP), hormone-sensitive lipase (HSL) genes and growth traits were studied in order to provide a theoretical basis for the two candidate genes in expression regulation in the yak. 【Method】Maiwa yak and partial sequences of H-FABP and HSL genes were used as experimental samples and candidate genes. The genetic diversity of Maiwa yak was studied by PCR-SSCP, and the alleles frequency, genotype frequency, equilibrium analysis of Hardy-Weinberg, heterozygosity (H), effective numbers of alleles (Ne) and polymorphism information content (PIC) by means of statistics method. The correlation between the candidate genes in different genotypes and the circumference of growth traits was analyzed.【Result】Polymorphism of H-FABP and HSL genes of some exon portions in Maiwa yak were found. There are three genotypes. The fitness test showed that only HSL gene exon 8 on polymorphic loci in Hardy-Weinberg equilibrium state and the rest of the polymorphic loci were deviation. H-FABP gene exon7 loci existed T→C mutation of 7 339 bp which didn't result in change of amino acid. HSL gene exon7 loci existed in G→C mutation of 6 883 bp, A→G mutation of 6 816bp which led to amino acid from D to G. Analysis indicated that interaction effect of the polymorphism loci genotype and growth traits was significantly (P<0.01). And A→G mutation of 10 183 bp in exon 8 which led to amino acid from C to G.【Conclusion】H-FABP and HSL genes in Maiwa yak have genetic polymorphism. A6816G genotype can be used as molecular marker.
    Construction of MyoD1 Expressed Goat Embryonic Fibroblast Line and the Myogenesis of This Cell
    LI Wei, ZHENG Meng-Meng, ZHANG Ya-Ni, ZHU Cai-Ye, QIU Feng-Long, WEI Guang-Hui, LI Bi-Chun
    Scientia Agricultura Sinica. 2013, 46(14):  3032-3039.  doi:10.3864/j.issn.0578-1752.2013.14.019
    Abstract ( 461 )   PDF (884KB) ( 683 )   Save
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    【Objective】 MyoD1 ectopic expression goat fibroblast line was constructed to explore the bio-functions of MyoD1 in myogenesis. 【Method】 MyoD1 was cloned from the skeletal satellite cell via RT-PCR method. To visualize the MyoD1 protein, MyoD1 was reconstructed to pEGFP-N1 with the mutation of TGA (termination codon) into GGA (glycine) and cloned into pEGFP-N1, a fusion protein expression vector. The MyoD1 expressed goat embryonic fibroblast (GEF) was founded to explore the function of MyoD1 during the myogenesis, via transfecting the pEGFP-MyoD vector into GEF with the help of LipofectiminTM LTX. MyoD1 expressed GEF was induced to undergo the myogenesis by the differentiation media. The myogenesis of this cell line was ditermined by morphology and indirect immunofluorescence assay (IFA). 【Result】 The sequencing results indicated that MyoD1 was successfully reconstructed into pEGFP-N1 with mutation of TGA into GGA, and MyoD1 was fused with eGFP. MyoD1 expressed GEF line was obtained through two weeks of continuous G418 resistance screening and expressed Myf5 (detected by indirect immunofluorescence assay (IFA)). A few signs of myotube were observed 2 days after differentiation treatment, which expressed the differentiation marker of myoblast (MyoG, Desmin, MyHC), while lots of myotube were obtained with contractility. 【Conclusion】MyoD1 gene was cloned and reconstructed with pEGFG-N1 to obtain pEGFP-MyoD1 expressed GEF line, successfully. The expression of MyoD1 would drive the GEF into myoblast with differentiation ability of myogenesis.
    Protection of Musovy Duck Reovirus by Immunization with the Recombinant σC and σB Protein Expressed in E. coli
    WU Xiao-Ping, WU Yi-Jian, HAN Dian-Lin, WU Bao-Cheng, HUANG Yi-Fan
    Scientia Agricultura Sinica. 2013, 46(14):  3040-3045.  doi:10.3864/j.issn.0578-1752.2013.14.020
    Abstract ( 494 )   PDF (622KB) ( 370 )   Save
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    【Objective】 The immunogenicity of the recombinant Musovy Duck Reovirus (DRV) capsid proteins (σC and σB) expressed in E. coli was investigated and thus the base of the research of effective subunit vaccine for DRV was established. 【Method】The stability of the recombinant E.coli pET-30a-S3/BL21 and pET-30a-S4 ORF2/BL21 were detected, while the recombinant proteins were emulsified after purification and renaturation. Fifty young musovy ducks were randomly divided into 5 groups including 3 treatment groups, the PBS control group and the infection control group. The treatment groups were injected subcutaneously with a dosage of 500 μg per injection, on 1 d and 7 d respectively. The recombinant DRV σB protein, the recombinant DRV σC protein, as well as both σB and σC proteins were used. On the 14d, the animals except the PBS control group, were challenged with YB isolate of DRV. The titer of the antibody raised by the recombinant proteins and the protection rate were detected. 【Result】The recombination E.coli was stable. The titers of the antibody of the immunity groups were all raised on the 7th day after the second injection. It was found that the titer of antibody in the group injected with both σB and σC proteins as well as the group injected with recombinant σC was 1﹕200, but in the meanwhile, that in the group injected with recombinant σB protein was just 1﹕100. The protection test showed that the injection with both σB and σC proteins could provide best protection for DRV infection with protection index 70, and then the second was the group injected with DRV σC protein with 60, while the last was the group with DRV σB protein with 40. 【Conclusion】The injection with both recombinant σB and σC protein expressed in E.coli and emulsified could provide protection to young musovy ducks resisting DRV infection.
    RESEARCH NOTES
    Differentially Expressed Protein Profile of Maize Seedling Leaves Between Hybrid and Its Parental Lines
    GUO Bao-Jian, SUI Zhi-Peng, LI Yang-Yang, FENG Wan-Jun, YAN Wen-Wen, LI Hui-Min, SUN Qi-Xin, NI Zhong-Fu
    Scientia Agricultura Sinica. 2013, 46(14):  3046-3054.  doi:10.3864/j.issn.0578-1752.2013.14.021
    Abstract ( 512 )   PDF (675KB) ( 799 )   Save
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    【Objective】To construct protein expression profile of seedling leaves between maize hybrid and its parental lines, with the purpose to give an insight into the molecular basis of leaf size heterosis. 【Method】 Differentially expressed protein profile of the third leaf on 5th day after germination between maize hybrid and its parental lines used for 2-DE analysis and differentially expressed proteins were identified by using MALDI TOF MS method. 【Result】 A total of 630 protein spots were detected, among which 52 protein spots were found to be differentially expressed between maize hybrid and its parental lines. Differentially expressed protein spots could be grouped into six models, that is UPF1 (expression in hybrid and uniparent but not in another parent), HDH(hybrid is equal to the highly expressed parent), LDH (hybrid is equal to the lowly expressed parent), URH (up-regulated in hybrid), DRH (down-regulated in hybrid), F1nBP (hybrid-specific expressed protein spots), and the numbers were 15, 13, 8, 6, 7 and 3, respectively. Moreover, 28 of the 52 differentially expressed protein spots were identified by using MALDI TOF MS, which were grouped into nine functional categories, including metabolism, development, stress, glycolysis, transcription regulation, protein folding and degradation, tricarboxylic acid cycle, cytoskeleton and unknown protein. 【Conclusion】 Significant alterations in protein expression occurred in seedling leaves of hybrids and parental lines and differentially expressed proteins implicated in some functional categories might contribute to heterosis related to leaf size heterosis.
    Effects of Deep Vertically Rotary Tillage on Grain Filling and Yield of Spring Maize in North Huang-Huai-Hai Region
    LI Yi-Bing-1, PANG Huan-Cheng-1, LI Hua-1, LI Yu-Yi-1, YANG Xue-1, DONG Guo-Hao-2, GUO Liang-Hai-2, WANG Xiang-Jun-2
    Scientia Agricultura Sinica. 2013, 46(14):  3055-3064.  doi:10.3864/j.issn.0578-1752.2013.14.022
    Abstract ( 488 )   PDF (2810KB) ( 726 )   Save
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    【Objective】Annual rotary tillage often can create a compacted plough pan in shallow soil layers, and has become a key factor hampering high crop yield in some main grain producing areas of China. In this study, therefore, a brand-new deep soil tillage called deep vertically rotary tillage was implemented to test its effects on grain filling and spring maize yield in north Huang-Huai-Hai region. 【Method】Tillage treatments including deep vertically rotary tillage of 30 cm (F30), and 50 cm (F50), deep vertically rotary tillage of 30 cm with film mulching (F30M) and 50 cm with film mulching (F50M) were conducted. Meanwhile, rotary tillage (XG) and subsoiling (SS) were used as experimental control. 【Result】Results showed that compared with rotary tillage and subsoiling, water infiltration under deep vertically rotary tillage got more easily, soil water storage was increased, soil water supply was improved, thus ear kernel number and kernel moisture were increased. Although the treatments of deep vertically rotary tillage had no advantage during the first stage of grain filling called slight-increase period (t1), while both initial grain-filling potential (R0) and mean grain-filling rate were decreased with the increase of tillage depth. Mean grain-filling rate of fast-increase period (the second stage, t2) and slow-increase period (the last stage, t3), on the other hand, was improved with the increase of tillage depth, i.e., F50 (50cm)>SS (35cm) >F30 (30cm) >XG (15cm). Finally, 100-kernel weight and yield of F30 and F50 were significantly increased compared with the rotary tillage and subsoiling. Due to moisture preservation of plastic film, ear kernel number was significantly increased compared with the treatments of F30 and F50. Furthermore, the grain filling of F30M and F50M showed an obvious superiority over rotary tillage and subsoiling from the beginning of t1. R0 and mean grain-filling rate of t1 of F30M and F50M were improved with the increase of tillage depth. Plus longer grain-filling period, 100-kernel weight and yield of F30M and F50M increased significantly compared with F30 and F50. 【Conclusion】The results showed that deep vertically rotary tillage with film mulching can provide a new and effective way to break up the compacted plough pan, solve the shallow tillage layer problem and increase crop yield.
    Regulatory Effects of Exogenous Nitric Oxide on Physiological Growth of Cotton Seedlings Under Nitrogen Stress
    CHEN Jing, LIU Lian-Tao, SUN Hong-Chun, ZHANG Yong-Jiang, WANG Zhan-Biao, LI Cun-Dong
    Scientia Agricultura Sinica. 2013, 46(14):  3065-3071.  doi:10.3864/j.issn.0578-1752.2013.14.023
    Abstract ( 396 )   PDF (506KB) ( 611 )   Save
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    【Objective】In order to provide a theoretical basis for regulation of SNP in cotton, the effects of NO donor sodium nitroprusside(SNP) on physiological growth of cotton seedlings were investigated under nitrogen stress. 【Method】 Two control groups were carried out in hydroponic experiment. The first control group cotton seedlings (CK1) were grown with Hoagland's solution. The second control group cotton seedlings (CK2) were grown with Hoagland's solution without nitrogen. Five treatment groups were designed under different SNP concentrations using as the basis. The concentrations of SNP, were 50, 100, 200, 500 and 1 000 μmol•L-1, respectively. The growth and leaves physiological characteristics of cotton seedlings under nitrogen stress were measured.【Result】Nitrogen stress inhibited the growth of cotton seedlings. Nitrogen stress changed the photosynthetic system and material content. Low concentration exogenous nitric oxide could increase cotton seedlings growth, increase the content of chlorophyll, proline and soluble protein significantly. The effect of nitrogen stress was relieved by lower concentration of NO (SNP: 100 μmol•L-1). The effect of nitrogen stress was enhanced by high concentration of NO. 【Conclusion】 Lower concentration of NO (SNP: 50-100 μmol•L-1) has a best effect on mitigating cotton seedlings’ nitrogen stress and can increase the tolerance of cotton nitrogen stress.
    The Optimization of Activated Carbon Decolorization Process of Preserved Ginger Syrup-Juice by Response Surface Methodology
    ZHANG Lu, QIAO Xu-Guang, LIU Xiao-Yu, YU Hua-Yang, DIAO Shun-Ze, TANG Xiao-Zhen
    Scientia Agricultura Sinica. 2013, 46(14):  3072-3078.  doi:10.3864/j.issn.0578-1752.2013.14.024
    Abstract ( 598 )   PDF (587KB) ( 573 )   Save
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    【Objective】The problem of preserved ginger syrup-juice became darker in color was solved to make sure the syrup-juice can be reused in preserved ginger production.【Method】Decolorization rate was used as an indicator in the experiment, UV scan was used to determine the max absorption wavelength of preserved ginger syrup-juice, single factor text and response surface methodology was used to get the optimal conditions, the single factor experiment was carried out with the activated carbon added at 1%, 2%, 3%, 4%, and 5%, temperature at 60℃, 70℃, 80℃, 90℃, and 100℃, and time at 10, 20, 30, 40, and 50 min, respectively.【Result】The max absorption wavelength of preserved ginger syrup-juice was 306nm, the optimal conditions of syrup-juice clarification using activated carbon were: the activated carbon added at 5%, the temperature at 79℃ and time was 35 min, and the decolorization rate could reach 45.1%.【Conclusion】The results of the experiment can be used as a technological support for reusing syrup-juice in the production of preserved ginger.