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    01 January 2012, Volume 45 Issue 1
    SPECIAL TOPICS
    Agriculture and Low-Carbon Development Strategy in China
    QI Ye, LI Hui-Min, WANG Xiao
    Scientia Agricultura Sinica. 2012, 45(1):  1-6.  doi:10.3864/j.issn.0578-1752.2012.01.001
    Abstract ( 742 )   PDF (340KB) ( 1226 )   Save
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    Carbon emission is the major cause of global climate change and a seriously constraint for sustainable development. Agriculture, characterized by its low emission, high carbon productivity and high sequestration capacity, is uniquely positioned with tremendous competitive advantage for low carbon development and thus should be reconsidered as a strategic industry in national development. At the national level, agriculture plays an important role in low carbon development as a low-carbon, ecological, and technological intensive industry. Low-carbon agriculture would not only help improve environmental quality but should also enhance food productivity and quality. The renewable energy production and carbon capture, use and storage (CCUS) application in agriculture can make a great contribution to the objective of national low-carbon development and the benefit of rural economy.
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Specific Binding of Wheat WPBF and Prolamin-Likebox in Upstream of HMW-GS Genes
    WEI Xiao-Bin, WANG Ya-Nan, ZHANG Chao, DAN Li-Wei, TANG Ru-Chun, FAN San-Hong
    Scientia Agricultura Sinica. 2012, 45(1):  7-15.  doi:10.3864/j.issn.0578-1752.2012.01.002
    Abstract ( 764 )   PDF (738KB) ( 1255 )   Save
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    【Objective】 The endosperm-specific transcription factor gene WPBF was cloned from wheat (Triticum aestivum L.), an optimized prokaryotic expression system was established, and the specific binding between WPBF and Prolamin-Like boxes in upstream of HMW-GS genes was verified. These will be useful for further elucidating the regulation mechanism of HMW-GS. 【Method】 The coding region of WPBF was obtained by RT-PCR and inserted into the vector pET-21a-MBP. The recombinant vector was expressed in E. coli strains T7 Express and Origami B(DE3), the recombinant protein was purified by amylose and Ni-NTA affinity chromatography, and its DNA binding activity was analyzed by electrophoretic mobility shift assay (EMSA).【Result】 Recombinant WPBF existed mainly in soluble form when it was expressed at 33℃ in Origami B (DE3) strain, and the fusion protein accounted for 48% of the total cell protein. The purified recombinant protein could bind specifically to two kinds of Prolamin-Like boxes,however,the binding strength between WPBF and Prolamin-Like box “TGCAAAG” was higher than WPBF and Prolamin-Like box“TGCAAG”. 【Conclusion】 An optimized prokaryotic expression system was established and the functional recombinant protein was obtained. The recombinant WPBF can bind specifically to two kinds of Prolamin-Like boxes in upstream of HMW-GS, and there is somewhat different in binding strength.
    Origin and Evolution of Jute Analysed by SRAP and ISSR Methods
    TAO Ai-Fen, QI Jian-Min, LI Mu-Lan, FANG Ping-Ping, LIN Li-Hui, XU Jian-Tang
    Scientia Agricultura Sinica. 2012, 45(1):  16-25.  doi:10.3864/j.issn.0578-1752.2012.01.003
    Abstract ( 955 )   PDF (372KB) ( 732 )   Save
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    【Objective】The objective of this study is to make clear the origin and evolution of Corchorus with two molecular methods (SRAP and ISSR). 【Method】Ninety-six jute germplasms were analysed with sequence related amplified polymorphism (SRAP) combining with inter-simple sequence related (ISSR) method. The phylogenetic trees of Corchorus were constructed by MEGA and DPS software, and the divergence time of jute germplasm was calculated. 【Result】 The relative wild species located at the basic position of the dendrogram, and the divergence time of which was the longest, which indicated the relative wild species originated earliest and was the ancestors of cultivated jute. Africa was the center of origin of Corchorus, while China was the second original center of Corchorus. Among all the olitorius species, the divergence time of wild and cultivated species from Africa was the longest, so Africa was the center of origin for wild and cultivated olitorius species, while the areas which border on India, Burma and China was the second center of origin for cultivated olitorius species. South China, countries of South Asia and Southeast Asia which border on China were the origin center of wild capsularis species, and South region of China was the origin center of cultivated capsularis species. Taken together, the divergence time of the cultivated capsularis species was shorter than olitorius species, indicating this biotype originated later than olitorius species. 【Conclusion】 Africa has an important position in origin of Corchorus, which was the primary origin center of wild species, wild and cultivated olitorius species. South region of China was the origin center of cultivated capsularis species. The divergence time and phylogenetic trees of Corchorus were calculated more comprehensively by SRAP markers combining with ISSR molecular markers, and the scientific conclusion on origin and evolution of Corchorus was got with above methods.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Soil Macro-structure and Precision Tillage
    DING Qi-Shuo, DING Wei-Min, PAN Gen-Xing, JI Chang-Ying
    Scientia Agricultura Sinica. 2012, 45(1):  26-33.  doi:10.3864/j.issn.0578-1752.2012.01.004
    Abstract ( 805 )   PDF (266KB) ( 619 )   Save
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    Soil macro-structure, an extrapolation from soil structure, is 1-2 magnitudes higher in scale than soil aggregates and  more spatially and temporally dynamic in nature. Soil physical composition, mechanical indices, mechanical governing mechanisms and potential mechanical applications in agricultural production comprised the key focus of soil macro-structure study, formulated a theoretical basis for precision tillage research. Indices for soil macro-structure quantification are comprised of shear strength, cone index, plate sinkage and fracture strength, which are governed by elasto-plastic, fracture and fragmentation, and rheological mechanics of soil macro-structural dynamics. Precision tillage is the effort of testing, evaluating and modeling the soil macro-structure. This quantification of the relationship between soil macro-structure dynamics and the mode and magnitude of the externally applied mechanical forces will finally lead to technical breakthroughs for precise placement or management of soil structure, crop residue, seed, water and fertilizer. The result of which will facilitate upgrading soil workability, enhancing soil structural stability, improving trafficability and soil bearing capacity, and optimizing soil-plant-environment or terra-machinery systems.
    Transfer Characteristics of Canopy Photo-Synthetically Active Radiation in Different Rice Cultivars Under Different Cultural Conditions
    ZHU Xiang-Cheng, TANG Liang, ZHANG Wen-Yu, CAO Meng-Ying, CAO Wei-Xing, ZHU Yan
    Scientia Agricultura Sinica. 2012, 45(1):  34-43.  doi:10.3864/j.issn.0578-1752.2012.01.005
    Abstract ( 761 )   PDF (558KB) ( 872 )   Save
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    【Objective】 The objective of this study was to explore the dynamic transfer characteristics of canopy photosynthetically active radiation (PAR) in different rice cultivars and under different cultural conditions.【Method】Field experiments were conducted in Nanjing in 2009 and in Yizheng in 2010, involving two cultivars with different plant types, three planting densities and five nitrogen rates. During rice growing periods, the dynamics of canopy structure and the daily changes in PAR transfer parameters were measured, and the effects of proportion of diffuse radiation on canopy PAR transfer characteristics were analyzed. 【Result】 The results indicated that canopy leaf area index (LAI), average leaf inclination and plant height of rice were affected significantly by planting densities and nitrogen rates. Dynamics of canopy PAR transmittance and reflectance firstly decreased and then increased over growing progress of rice, with the minimum value appeared during booting to heading stages. Canopy PAR reflectance decreased with enhancing nitrogen rates. Extinction coefficient (K) in canopy increased over growing progress of rice and with increasing planting densities and nitrogen rates. The dynamics of K with the days after transplanting could be described with the upward exponential equation. The daily change patterns of canopy PAR reflectance and interceptance, and K values could be described with downward parabolic curves, reaching the minimum values at noon, while opposite patterns were seen /in the PAR transmittance. The ranges of daily variation in canopy PAR transmittance and interceptance, and K values at grain filling stage were obviously lower than those at tillering stage. The dynamics of K with the solar angle could be described with the Doseresp curve, which was affected by the canopy structure and cultivar genetic characteristics. Canopy PAR transmittance decreased and K value enhanced with reduced proportion of diffuse radiation.【Conclusion】The dynamic characteristics of canopy PAR transfer in rice were regulated by planting densities and nitrogen rates, exhibiting marked changes over daily times and growth stages, and affected by the proportion of diffuse radiation. These results will provide experimental supports for precision simulation of canopy photosynthesis and productivity in rice.
    Effect of Cytokinins on the Growth of Rice Tiller Buds and the Expression of the Genes Regulating Rice Tillering
    LIU Yang, GU Dan-Dan, XU Jun-Xu, DING Yan-Feng, WANG Qiang-Sheng, LI Gang-Hua, LIU Zheng-Hui, WANG Shao-Hua
    Scientia Agricultura Sinica. 2012, 45(1):  44-51.  doi:10.3864/j.issn.0578-1752.2012.01.006
    Abstract ( 1102 )   PDF (396KB) ( 1258 )   Save
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    【Objective】 The objective of the present study was to investigate the relationship of cytokinins (CTK) and the expression of the genes regulating rice tillering during the regulating the growth of rice tiller buds growth and the underlying molecular mechanisms. 【Method】 In the present study, a cultivar Nanjing 44 was used. At tillering stage, 40 mg?L-1 nitrogen (N) was used to stimulate the germination of tiller buds. At the same time, external IAA was used to inhibit the germination of tiller buds. The changes of CTK contents in tiller buds and tiller nodes and the expression of OsTB1, OsD3, OsD10 and OsD27 in tiller buds were measured. 【Result】 The external 40 mg?L-1 N increased the CTK contents in tiller nodes and tiller buds, inhibited the expression of OsTB1, OsD3, OsD10 and OsD27 in tiller buds, and then promoted the growth of rice tiller buds. However, the external application of IAA reversed the effect of N. Besides this, the expression of OsTB1 in tiller buds was regulated by CTK, however, the expression of OsD3, OsD10 and OsD27 in tiller buds might not be regulated by CTK. 【Conclusion】 The results showed that external N and IAA might have regulated tiller bud growth by at least two means: one is regulating the CTK content and the expression level of OsTB1 in tiller buds, another is regulating the expression level of OsD3, OsD10 and OsD27 in tiller buds and by this to regulate the tiller bud growth.
    PLANT PROTECTION
    SSR Marker Development and Analysis of Genetic Diversity of Fusarium verticillioides Isolated from Maize in China
    REN Xu, ZHU Zhen-Dong, LI Hong-Jie, DUAN Can-Xing, WANG Xiao-Ming
    Scientia Agricultura Sinica. 2012, 45(1):  52-66.  doi:10.3864/j.issn.0578-1752.2012.01.007
    Abstract ( 1096 )   PDF (774KB) ( 1304 )   Save
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    【Objective】 The objective of this study is to analyze genetic diversity of Fusarium verticillioides and to develop SSR markers for this fungus. 【Method】 Fastpcr was used for searching SSRs in genomic sequence of F. verticillioides. The sequences including suitable SSRs were selected to design primers using Primer5.0. The NTSYS and Popgene programs were used to analyze genetic diversity of maize isolates of F. verticillioides. 【Result】 Among the 158 primer pairs designed, 109 pairs (69.0%) were able to amplify 2 or more alleles against discretionary F. verticillioides isolates originating from single conidium. Of them 55 pairs (34.8%) amplified 3 or more alleles. In 11 chromosomes assembled 22 primer pairs, two pairs for one chromosome, were selected for genetic diversity analysis of 66 isolates of F. verticillioides from maize. In 22 SSR loci, a total of 125 alleles (2 to 11 alleles amplyfied by per primer pair) were detected with an average of 5.68 alleles. The Nei’s gene diversity, ranged from 0.1139 to 0.8687 with an average mean of 0.6199, indicated high genetic diversity in F. verticillioides isolates from various geographic regions.【Conclusion】Based on genome information of F. verticillioides, 109 SSR markers were developed and showed high polymorphism and high diversity among F. verticillioides isolates. By amplyfying with 22 SSR primer pairs, 66 isolates of F. verticillioides from maize were seperated into 3 groups on 0.3 coefficient. The relationship between genetic diversity and geographical distribution did not show correlation in isolates of F. verticillioides from maize in China.
    Study on Ecological Niches of Main Predatory Arthropods in Integrated Rice-Duck Farming System
    QIN Zhong, ZHANG Jia-en, ZHANG Jin, LUO Shi-Ming
    Scientia Agricultura Sinica. 2012, 45(1):  67-76.  doi:10.3864/j.issn.0578-1752.2012.01.008
    Abstract ( 702 )   PDF (328KB) ( 538 )   Save
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    【Objective】The objective of this study is to quantify resource utilization of main predatory arthropods in integrated rice-duck farming system and to explore their inner relationship in resource allocation. 【Method】Through field investigations on main predatory arthropods in the integrated rice-duck farming system and conventional rice cultivation system, the structure of the predatory arthropods communities and their niches were explored. Meanwhile, the quantitative distribution characters of each group in resource sequence and the condition of resource utilization were analyzed. 【Result】Dominant species of predatory arthropods in the integrated rice-duck farming system and conventional rice cultivation system were Erigonidae, Araneidae, Lycosldae and Tetragnathidae, in which the dominance index of Araneidae was the greatest, accounted for 0.4999 and 0.6665, respectively, in two rice cultivation systems. Foraging ducks in rice cropping field directly or indirectly influenced the number and distribution pattern of main predatory arthropods, thus leading to various degrees of changes in temporal-spatial dimensional niche breadth, niche overlap index and the niche proportional similarity. Temporal-spatial dimensions niche breadth of Erigonidae, Araneidae and Tetragnathidae were lower than those in the conventional rice cultivation system. The niche overlap indices of Erigonidae and Theridlidae, Tetragnathidae and Salticidae, Salticidae and Clubionidae, Clubionidae and Coccinellidae in the integrated rice-duck farming system were 70.65%-75.57% higher than those in the conventional rice cultivation system. 【Conclusion】Although the total individual numbers of predatory arthropods in the rice-duck farming system was 19.56% lower than those in the conventional rice cultivation system, group competition in the same resource sequence among main predatory arthropods became more intensified and time synchrony in resource utilization was more pronounced, which meant that predatory arthropods could make the best use of more potential resources in a more uniformed distribution and have a stronger controlling effect on pest population by longer occurrence time and continuous controls.
    Insecticidal Activities and Active Ingredients of Pronephrium megacuspe
    HUANG Su-Qing, XU Han-Hong, TIAN Yong-Qing, WEI Xiao-Yi
    Scientia Agricultura Sinica. 2012, 45(1):  77-84.  doi:10.3864/j.issn.0578-1752.2012.01.009
    Abstract ( 703 )   PDF (358KB) ( 531 )   Save
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    【Objective】 The objective of this study is to research the active ingredients of Pronephrium megacuspe.【Method】Insecticidal activities and action mode were determined by bioassay. The active ingredients were isolated from the extract by activity-guided fractionation with column chromatography, thin layer chromatography (TLC) and recrystallization, and the structures were identified predominantly on the basis of MS and NMR data.【Result】The results showed that the methanol extract of P. megacuspe possessed toxicity against the adult of Musca domestica, the 4th instar larva of Aedes albopictus and Myzus persicae, and had antifeedant activity to the 3rd instar lavae of Plutella xylostella, Ostrinia furnacalis and Pieris rapae. 7-hydroxy-5-methoxy -6,8-dimethyl-flavanone, 2′,4′-dihydroxy-3′-methyl-6′-methoxychalcone, 5,7-dihydroxy-6,8-dimethylflavanone and 5-hydroxy-6,7- dimethoxyflavone were isolated and identified from the methanol extract of P. megacuspe. The results of bioassay showed that all compounds possessed antifeedant activity to the 3rd instar lavae of O. furnacalis with the average antifeedant rate above 50% at a concentration of 50 mg•L-1 at 24 h after treatment. Four compounds exhibited no contact activity against the 3rd instar lavae of O. furnacalis at a dosage of 2 ?g/pest. 5,7-dihydroxy-6,8-dimethylflavanone possessed weak antifeedant activity against the 3rd instar lavae of P. xylostella with an average antifeedant rate of 40.51% at 24 h after treatment, and the mortality was 33.76% at a dosage of 1 ?g/pest at 48 h after treatment. The other three compounds had no contact toxicity to the 3rd instar lavae of P. xylostella. 【Conclusion】P. megacuspe has insecticidal activities against various insect pests. Four compounds were isolated and identified from the methanol extract of P. megacuspe. They have the similar mother structure, but different on the number of replaced methylium and its place, and have different activities. Therefore the other compounds and the structure-activity relationship of P. megacuspe need to be further investigated as a biopesticide.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Influence of Water Supply and Phosphorus Application in Different Depth on Growth and Water Use Efficiency of Winter Wheat
    KANG Li-Yun, LI Shi-Qing
    Scientia Agricultura Sinica. 2012, 45(1):  85-92.  doi:10.3864/j.issn.0578-1752.2012.01.010
    Abstract ( 747 )   PDF (297KB) ( 746 )   Save
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    【Objective】Experiments on effects of water supply and phosphorus application in different depths on root distribution and grain yield of winter wheat are of theoretical and practical importance to phosphorus and growth in dryland farming. 【Method】A cylindrical pot experiment was conducted with Eum-Ortllic Anthrosols (Cinnamon soils) to study the effects of water supply and phosphorus application in different depths on root distribution, grain yield and water use efficiency (WUE) of winter wheat. There were three phosphorus levels including no phosphorus (CK) and P was placed either in the topsoil at 0-30 cm (UP) or in the subsoil at 30-60 cm (LP). Water treatments were introduced to provide ample water supply with both topsoil and subsoil wet (W) and dry topsoil but ample water supply in the subsoil (D). 【Result】 The results showed that there were large difference in root distribution, yield and WUE of wheat among different treatments. Compared with W treatment, D treatment reduced 0-30 cm root biomass by 19.6%, but the 30-60 cm root biomass increased by 18.8%. The phosphorus treatments affected the distribution of roots more than total root biomass. Root biomass was increased at the site of placement where phosphorus was applied, and placement of phosphorus at 30-60 cm increased with deep placement compared to placement at 0-30 cm under water treatment of D condition. The different water treatments significantly affected yield and WUE of wheat (P<0.01). Phosphorus application caused higher production and WUE, increased by 10.0% and 47.4%, respectively. The placement of fertilizer P on them played different roles under two water treatments. Under D condition, placement of phosphorus at 30-60 cm produced higher yields and WUE than placement at 0-30 cm, increased by 11.2% and 28.6%; on the contrary, placement of phosphorus at 30-60 cm decreased yields and WUE by 41.1% and 37.9% under ample water supply with both topsoil and subsoil wet. 【Conclusion】 The simulated study suggested that deep placement of fertilizer phosphorus caused greater root growth, better water use efficiency and higher production under two water conditions, especially in dry topsoil but ample water supply in the subsoil .
    Study on Index System of Optimal Fertilizer Recommendation for Spring Corn in Hetao Irrigation Area of Inner Mongolia
    LI Wen-Biao, LIU Rong-Le, ZHENG Hai-Chun, LI Shu-Tian, GAO Fan-Shen, LIU Ji-Pei
    Scientia Agricultura Sinica. 2012, 45(1):  93-101.  doi:10.3864/j.issn.0578-1752.2012.01.011
    Abstract ( 776 )   PDF (381KB) ( 642 )   Save
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    【Objective】 The index system of optimal fertilizer recommendation for scientific fertilization of spring corn in hetao irrigation area was established in this study. 【Method】 “3414” fertilizer efficiency Experiments on spring corn were designed based on high, midle and low soil fertility in Hetao irrigation area. 【Result】The results showed that agronomic efficiency of N, P2O5 and K2O on spring corn was 10.6 kg•kg-1, 9.9 kg•kg-1 and 6.7 kg•kg-1, respectively, with the role of yield increasing effect of N>P2O5>K2O. The rate of yeild increasing by fertilizers increased with the declining of soil fertility, while yeild increase per unit nutrient decreased. The indices of abundance and deficiency of soil nutrients in Hetao irrigation area were showed as follow: total N with five levels of lower (<0.42 g•kg-1), low (0.42-0.67 g•kg-1), moderate (0.67-1.08 g•kg-1), high (1.08-1.72 g•kg-1), and higher (>1.72 g•kg-1); Olsen-P with five levels of lower (<3.6 mg•kg-1), low (3.6-7.6 mg•kg-1), moderate (7.6-16.0 mg•kg-1), high (16.0-33.7 mg•kg-1), and higher (>33.7 mg•kg-1); Solubale K with five levels of lower (<70 mg•kg-1), low (70-103 mg•kg-1), moderate (103-151 mg•kg-1), high (151-222 mg•kg-1), and higher (>222 mg•kg-1). The fertilization model of spring corn in Hetao irrigation area was F(N)=-78.13×lnS(N)+155.85, F(P2O5)=-54.621×lnS(P)+250.2, F(K2O)=-42.4071×lnS(K)+272.91. 【Conclusion】 Agronomic efficiency, fertilizer efficiency, abundance and deficiency indices of soil nutrients and fertilization model were important bases for optimal fertilizer recommendation.
    HORTICULTURE
    Cloning and Expression of the Sorbitol Dehydrogenase Gene Family in Apple
    LIANG Dong, WU Shan, WANG Su-Fang, MA Feng-Wang
    Scientia Agricultura Sinica. 2012, 45(1):  102-110.  doi:10.3864/j.issn.0578-1752.2012.01.012
    Abstract ( 978 )   PDF (400KB) ( 863 )   Save
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    【Objective】 The objective of this experiment was to further investigate the molecular characteristics of sorbitol dehydrogenase (SDH) in apple. 【Method】 The cDNAs and gDNAs of SDH gene family were isolated from the fruit of Malus domestica cv. Gala, and the mRNA expression levels of SDH gene family during leaf and fruit development were studied. 【Result】Twelve cDNAs of SDH were cloned from apple fruit, in turn being named MdSDH7-MdSDH18. Intron/exon structure analysis indicated that MdSDH18 contained 6 extons and 5 introns, whereas others contained 2 extons and 1 intron. The mRNA expression levels and enzyme activities of SDH were detected during leaf and fruit developments. According to the amino acid sequence homology, these SDHs were divided into two groups, A and B. During the fruit development, the mRNA expression levels of all SDHs at middle phase and late phase were higher than that of early phase, however, the changes of SDH enzyme activity were opposite to gene expression patterns. During leaf development, mRNA expression levels of A-SDH were in the order of young leaf > senescence leaf > mature leaf, with activities showing similar patterns to gene expression. However, mRNA expression   levels of B-SDH was in the order of senescence leaf >mature leaf > young leaf. 【Conclusion】 These results further suggest that SDH is a gene family in apple, and the expression of gene family members shows different patterns during leaf and fruit developments.
    Changes of Water Transportation in Berry Vascular Bundle at Different Developmental Phases of Kyoho Grape Berry
    XIE Zhao-Sen, CAO Hong-Mei, LI Bo, LI Wei-Fu, XU Wen-Ping, WANG Shi-Ping
    Scientia Agricultura Sinica. 2012, 45(1):  111-117.  doi:10.3864/j.issn.0578-1752.2012.01.013
    Abstract ( 824 )   PDF (567KB) ( 1417 )   Save
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    【Objective】The objective of the experiment is to understand the relation ship between water transportation in berry vascular and berry growth.【Method】The berry growth, berry firmness and turgor, the structure of vascular bundle and water transportation in vascular bundle were studied. Apoplastic dye stuff was used as the tracer to study the water transportation in berry vascular.【Result】The results showed that during phase I, the dorsal and central vascular bundles were colored mostly, the speeds of dye stuff transport in dorsal vascular bundles were the highest among three phases of berry development, the speed was 0.97 cm·h-1. After phase II, the distribution of dye stuff decreased, the speeds of dye stuff transport decreased, the speeds in dorsal and central vascular bundles were 0.08 cm·h-1 and 0.72 cm·h-1, respectively. During phase III, the distribution of dye stuff was still lower than phase I. After phase II, the walls of xylem vessels were indistinct, some of them were broken.【Conclusion】After phase III, although the collapse of partial xylem structure and non-functional xylem led to the decrease in efficiency of water transportation in vascular, sugar accumulation in grape berry increased, osmoregulation in berry also increased, water entered into grape berry via phloem. These adjustments promoted the second rapid growth of grape berry.
    Molecular Cloning of Longan Caffeoyl-CoA O-methyltransferase (DLCCoAOMT) and Its Expression Analysis
    CHEN Hu, HE Xin-Hua, LUO Cong, YANG Li-Tao, ZHANG Bao-Qing, SONG Xiu-Peng
    Scientia Agricultura Sinica. 2012, 45(1):  118-126.  doi:10.3864/j.issn.0578-1752.2012.01.014
    Abstract ( 852 )   PDF (863KB) ( 834 )   Save
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    【Objective】The aim of this study was to clone full-length cDNA of a key enzyme gene DLCCoAOMT related to lignin metabolism in longan, investigate its sequence characteristics and analyze its expression in different organs under low temperature stress, as well as expression in the prokaryote. 【Method】 The DLCCoAOMT gene cDNA sequence was cloned from longan leaf using RT-PCR and RACE techniques. The bioinformatics methods were used to analyze putative amino acid sequence and real-time PCR method were used to study the expression of DLCCoAOMT gene in different tissues. 【Result】 The full-length cDNA of DLCCoAOMT (GenBank accession number: JN093023) in longan was cloned. The sequence consists of 993 bp with an open reading frame of 744 bp, encoding a polypeptide of 247 amino acids. Homology analysis showed that the deduced DLCCoAOMT protein was highly homologous to other CCoAOMT proteins from different species. Phylogenetic analysis also indicated that DLCCoAOMT was very closely related to DLCCoAOMT of Betula. Real-time PCR results showed that the DLCCoAOMT expressed in root, stem and leaf, and its expression was different among three organs. The mRNA of DLCCoAOMT was abundant in root and stem, but less in leaf. Furthermore, DLCCoAOMT transcription level was significantly different among root, stem and leaf with time courses of chilling treatment. Prokaryotic expression showed that recombinant plasmid was efficiently expressed in Escherichia coli BL21. 【Conclusion】 Gene DLCCoAOMT was firstly isolated and characterized from longan, which may be involved in chilling stress.
    Changes of Cell Wall Degrading Enzymes in Potato Tuber Tissue Slices Infected by Fusarium sulphureum
    YANG Zhi-Min, BI Yang, LI Yong-Cai, KOU Zong-Hong, BAO Gai-Hong, LIU Cheng-Kun, WANG Yi, WANG Di
    Scientia Agricultura Sinica. 2012, 45(1):  127-134.  doi:10.3864/j.issn.0578-1752.2012.01.015
    Abstract ( 1010 )   PDF (398KB) ( 974 )   Save
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    【Objective】The dynamic changes of main cell wall degrading enzymes (CWDEs) activities in potato tuber tissue slices infected by Fusarium sulphureum were investigated.【Method】Major CWDEs activities of potato tuber slices tissue (Longshu No.3) inoculated with F. sulphureum were detected and comparatively analyzed during different sampling period. 【Result】 A series of CWDEs including polygalacturonase (PG), polymethyl-galacturonase (PMG), carboxymethyl cellulose (Cx), β-glucosidase, polygalacturonic acid trans-eliminase (PGTE), pectin methyl-trans-eliminase (PMTE), pectin methylesterase (PE) and pectate lyase were detected in infected tissue. Whereas PG, PMG, Cx, and β-glucosidase showed higher activities as compared with others. The activity of PMG and Cx kept higher during early infection period (1-3 days after infection), but higher PG activity appeared during later infection period (4-6 days after inoculation), but β-glucosidase showed a rising trend during the enzyme dynamics analysis.【Conclusion】 These findings suggested that CWDEs were the primary pathogenic factors, and different CWDEs might take effect at different infecting periods.
    STORAGE·FRESH-KEEPING·PROCESSING
    EPR Spectroscopy Studies on DPPH Free Radical Scavenging Activity of Dry Red Wines and Their Relationship with Total Polyphenols Content
    HU Bo-Ran, WEN Wen, YIN Shu-Zhen, ZHU Yong, ZHANG Feng-Min
    Scientia Agricultura Sinica. 2012, 45(1):  135-141.  doi:10.3864/j.issn.0578-1752.2012.01.016
    Abstract ( 1011 )   PDF (461KB) ( 720 )   Save
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    【Objective】 The purpose of this study was to compare the antioxidant capacities of dry red wines and analyze their relationship with total polyphenol content. 【Method】 The antioxidant properties of five monoverietal dry red wine samples (from the Shacheng viticultural region of China,) were investigated by Electron Paramagnetic Resonance (EPR) spectroscopy using 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical as oxidants, and their total polyphenols content were analyzed by FC method. 【Result】Generally, the dry red wines which were diluted 200 folds showed high antioxidant in 5 min reaction time, the DPPH scavenging rate of Cabernet was 72.31% which was the highest, Ruby Cabernet 72.23%, Merlot 68.31%, Shiraz 59.03% and Zinfandel 56.34%;the total polyphenols content of Merlot was the highest at 3 044.43 mg•L-1, Ruby Cabernet 3 026.92 mg•L-1, Cabernet 2 886.81 mg•L-1, Shiraz 2 085.51 mg•L-1 and Zinfandel 1 647.64 mg•L-1. 【Conclusion】 It is an efficient method in detecting the antioxidant capacities of wines by EPR method. The DPPH scanvenging activity from dry red wines showed obviously difference. There was a linear correlation between the antioxidant activities and the total polyphenols content, y=0.01116x+ 37.327665, rGAE/DPPHequiv=0.91753.
    Regression Analysis of Sensory Characteristics and Volatile Compounds in Pork Product During Cold-Storage
    LI Hong-Jun, HUANG Ye-Chuan, HE Zhi-Fei, LI Feng
    Scientia Agricultura Sinica. 2012, 45(1):  142-152.  doi:10.3864/j.issn.0578-1752.2012.01.017
    Abstract ( 789 )   PDF (610KB) ( 593 )   Save
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    【Objective】 The objective of this study was to investigate the changes in volatile compounds and sensory characteristics, and the relationship between them in pork product during cold-storage. 【Method】 The longissimus muscles from Rongchang (RC) and PIC pigs were cooked at different temperatures respectively, and after keeping in the dark at 4℃ in oxygen permeable bags for 0, 3 or 6 days, the sensory characteristics, TBARS values and volatile compounds of all samples were analyzed. 【Result】During cold-storage, the concentration of some lipid-derived products, TBARS value and warmed-over flavor (WOF-F) significantly increased while the meat-like flavor (Meat-F) markedly decreased in samples. Analysis of partial least squares regression (PLS) showed that TBARS value, Meat-F and WOF-F score were significantly related to 8, 31 and 23 kinds of volatile compounds respectively, and regression models established on the basis of these volatile compounds could well predict TBARS value, Meat-F and WOF-F score in each sample. 【Conclusion】 The changes of sensory characteristics in pork products during cold-storage were mainly due to the concentration variation of some volatile compounds.
    ANIMAL SCIENCE·RESOURCE INSECT
    Advances in Research on Biodegradation of Mycotoxins in Feed
    JI Cheng
    Scientia Agricultura Sinica. 2012, 45(1):  153-158.  doi:10.3864/j.issn.0578-1752.2012.01.018
    Abstract ( 921 )   PDF (283KB) ( 1851 )   Save
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    Mycotoxins are extremely toxic and carcinogenic. They pose severe threat to both livestock productivity and human health and thus bring huge worldwide economic losses in animal husbandry and food industry annually. A safe, effective and environment friendly detoxification method is urgently needed for controlling these toxins because of many application defects of physical and chemical detoxification. Toxicological mechanism and biodegradation research of mycotoxins are comprehensively reviewed, and some strategies and advices on existing difficulties of biodegradation research are also proposed in this paper.
    Effect of Dietary Metabolizable Energy and Crude Protein Levels on Performance of Lingnan Yellow-Feathered Broiler Breeders
    ZHU Cui, JIANG Zong-Yong, JIANG Shou-Qun, ZHOU Gui-Lian, LIN Ying-Cai, CHEN Fang, ZHENG Chun-Tian, HONG Ping
    Scientia Agricultura Sinica. 2012, 45(1):  159-169.  doi:10.3864/j.issn.0578-1752.2012.01.019
    Abstract ( 806 )   PDF (418KB) ( 615 )   Save
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    【Objective】 This experiment was conducted to investigate different dietary levels of metabolizable energy and crude protein on performance of Lingnan Yellow-Feathered Broiler Breeders. 【Method】 A total of 1 134 30-week-old Lingnan Yellow-Feathered Broiler Breeders were randomly assigned into 3×3 factorial dietary treatments with 3 dietary metabolizable energy (ME) levels at 11.09, 11.51, and 11.92 MJ•kg-1 and 3 crude protein (CP) levels at 15.5%, 16.5%, and 17.5%. Each dietary treatment had 6 replicates with 21 birds per replicate. Experimental birds were fed 130 g of diet per bird every day. Growth performance, laying performance, and egg incubation indicators were determined. The trial lasted for 10 weeks.【Result】Broiler breeders fed       11.92 MJ•kg-1 ME diet had significantly greater weight gain and lower FCR (feed intake/egg mass ratio) when compared with birds of other ME groups (P<0.01). However, CP levels had no significant influence on weight gain and FCR of broiler breeders (P>0.05). An increase in dietary ME from 11.09 to 11.92 MJ•kg-1 of diet significantly improved egg production, average egg weight, and daily egg mass (P<0.05). Diet containing 16.5% CP or 17.5% CP significantly increased the average egg weight and reduced the egg breakage of broiler breeders (P<0.05) compared to 15.5% CP treatment. There was a significant interaction between ME and CP levels on unqualified egg percentage and hatchability (P<0.05). Chick birth weight was significantly improved by dietary 11.92 MJ•kg-1 ME and 16.5% CP treatments when compared to 11.09 MJ•kg-1 ME treatment and 15.5% CP treatment, repectively (P<0.05). 【Conclusion】The results indicated that the optimal dietary ME and CP level for Yellow-Feathered Broiler Breeders at 30-39-week of age was 11.92 MJ•kg-1 and 16.5%, respectively, whose ME and CP requirements as units per hen per day was estimated to be 1.55 MJ and 21.45 g, respectively.
    Effect of Controlled Freezing and Open-Pulled Straw (OPS) Vitrification on ATP Content and Reactive Oxygen Species (ROS) Level of Bovine In Vitro Produced Blastocysts
    ZHAO Xue-Ming, DU Wei-Hua, WANG Dong, HAO Hai-Sheng, ZHU Hua-Bin
    Scientia Agricultura Sinica. 2012, 45(1):  170-177.  doi:10.3864/j.issn.0578-1752.2012.01.020
    Abstract ( 914 )   PDF (348KB) ( 830 )   Save
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    【Objective】 The experiment was designed to investigate the effect of cryopreservation on ATP content and ROS level of bovine blastocysts produced by in vitro fertilization (IVF) and somatic cell nuclear transfer (SCNT). 【Method】 Bovine in vitro produced blastocysts were cryopreserved by controlled freezing and OPS vitrification. ATP Bioluminescence Assay Kit HS II and GENMED ROS Assay Kit were used to analyze ATP content and ROS level in cryopreserved blastocysts. 【Result】 (1) For each kind of blastocysts (IVF or SCNT), significant difference was observed between survival rates of controlled freezing group ((81.25±4.98)% or (49.41±2.24)%) and OPS vitrification group ((93.25±5.17)% or (77.56±3.52)%) (P<0.05). (2) For IVF blastocysts, ATP content was significantly decreased from (0.76±0.04) pmol (fresh group) to (0.45±0.03) pmol (controlled freezing group) or (0.63±0.05) pmol (vitrification group) (P<0.05), and the same was found for SCNT blastocysts ((0.40±0.02) pmol to (0.22±0.01) pmol or (0.33±0.02) pmol) (P<0.05). ATP content of each kind of blastocysts (IVF or SCNT) in controlled freezing group ((0.45±0.03) pmol or (0.22±0.01) pmol) was significantly lower than that in OPS vitrification group ((0.63±0.05) pmol or (0.33±0.02) pmol) (P<0.05). (3) ROS level of fresh IVF blastocysts ((48.52±2.65) cps) or SCNT blastocysts ((27.36±2.23) cps) was significantly lower than that of OPS vitrification group ((74.34±4.24) cps or (43.21±3.35) cps) (P<0.05), but higher than that of controlled freezing group ((35.61±4.32) cps or (16.56±2.52) cps) (P<0.05). 【Conclusion】 The present work indicated that vitrification was more efficient in the cryopreservation of bovine blastocysts derived from IVF or SCNT with comparison of controlled freezing. Meanwhile, both vitrification and controlled freezing significantly altered ATP content and ROS level in those blastocysts.
    VETERINARY SCIENCE
    Preparation of Polyclonal Anti-Sox2 Antibody in Capra hircus
    LIU Ping, ZHANG Yun, ZHENG Xi-Bang, LI Gong-He, CEN Xiao-Mei, YUE Lei-Lei, ZONG Zi-Jie, LU Sheng-Sheng, LU Ke-Huan, ZHANG Ming
    Scientia Agricultura Sinica. 2012, 45(1):  178-183.  doi:10.3864/j.issn.0578-1752.2012.01.021
    Abstract ( 826 )   PDF (467KB) ( 1203 )   Save
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    【Objective】 The present study was to construct a prokaryotic expression vector of Capra hircus Sox2 gene, pRSET-Sox2, to induce expression and purification of His-Sox2 fusion protein, which was used to immunize New Zeland white rabbits to prepare polyclonal anti-Sox2 antibody. 【Method】 Removed from plasmid pMD18-Sox2 by double digestion of BamH I and Xho I, Sox2 fragment was subcloned to pRSET-A vector to construct recombinant plasmid pRSET-Sox2. The plasmid was transformed into E. coli BL 21 (DE3), and His-Sox2 fusion protein was induced to expess with 1 mmol•L-1 IPTG at 37℃ for 4 h, which was identified with SDS-PAGE analysis and Western blotting. In the same way, large volume of expressing culture was prepared to purify His-Sox2 fusion protein with NI-NTA argrose under denaturing condition. The refolded fusion protein in vitro was injected subcutaneously into New Zeland white rabbits for four times at intervals of 2-3 weeks. Seven days after the last injection, blood samples were collected, serum was isolated, and specificity of polyclonal anti-Sox2 antibody was determined by Western blotting assay.【Result】 The prokaryotic expression vector pRSET-Sox2 was expressed efficiently in E. coli. BL21. The purified His-Sox2 was qualified for preparation of polyclonal antibody. The polyclonal anti-Sox2 antibody was prepared, and it could bind His-Sox2 fusion protein specifically, which was illustrated by Western blotting assay. 【Conclusion】 The polyclonal anti-Sox2 antibody with strong specificity was prepared, which will lay a solid biological foundation for study of Sox2, and for its application in detection of Capra hircus iPS cells (induced pluripotent stem cells).
    Expression of TRPV6 in Different Reproductive Organs in Laying Hens
    YANG Jun-Hua, HOU Jia-Fa, DENG Yi-Feng, ZHOU Zhen-Lei, WANG Yong-Mei, SHI Lu-Juan
    Scientia Agricultura Sinica. 2012, 45(1):  184-191.  doi:10.3864/j.issn.0578-1752.2012.01.022
    Abstract ( 811 )   PDF (594KB) ( 655 )   Save
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    【Objective】 The aim of this study was to investigate the localization and expression of transient receptor potential vanilloid receptor 6 (TRPV6) in the ovary, magnum, isthmus and uterus parts of the oviduct in laying hens. 【Method】Immunohistochemical technology and RT-PCR were used to detect the localization of TRPV6 in different reproductive organs. In addition, quantitation of TRPV6 mRNA level and protein concentration in these tissues were carried out by real-time PCR and Western blotting analysis, respectively. 【Result】 Positive expression of the TRPV6 protein and mRNA was observed in ovarium, magnum, isthmus and uterus parts of the oviduct in laying hens, respectively. TRPV6 was localized in the primary follicle and secondary follicle in the ovary. Furthermore, intense staining was observed in villus epithelial cell (face to lumen) of mucous epithelium of the magnum, isthmus and uterus parts of the oviduct. And the weaker staining was also found in the basilar layer of endometrium in the uterus parts of the oviduct. Moreover, the mRNA levels of TRPV6 in the magnum and isthmus were significantly decreased compared with that in the ovary (P<0.05). The expression of TRPV6 in the magnum was also lower than that in the ovary (P<0.05). However, the expression of TRPV6 mRNA in the uterus parts of the oviduct was lower than that in the ovary, but the change of protein concentration was in contrast, and the difference was not significant (P>0.05).【Conclusion】 The expression of TRPV6 was detected in ovarium, magnum, isthmus and uterus parts of the oviduct in laying hens. And the higher concentration of TRPV6 in ovary and uterus parts implied that the TRPV6 may be not only involved in the development and maturation, but also may play an important role in Ca2+ transport in ESG (eggshell gland) during eggshell calcification.
    RESEARCH NOTES
    Effect of Paclobutrazol on Starch Accumulation and Related Enzyme Activity of Storage Root in Edible Sweet Potato cv. Beijing 553
    CHEN Xiao-Guang, SHI Chun-Yu, WANG Zhen-Lin, ZHANG Li-Ming, ZHANG Xiao-Dong
    Scientia Agricultura Sinica. 2012, 45(1):  192-198.  doi:10.3864/j.issn.0578-1752.2012.01.023
    Abstract ( 920 )   PDF (353KB) ( 627 )   Save
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    【Objective】 The objective of this study was to discuss the effects of paclobutrazol on the physiological mechanism of promotion of the storage root growth rate and increase of storage root yield in edible sweet potato cv. Beijing 553.【Method】Field plot experiments with paclobutrazol at M0, 100 mg•kg-1 (M1), and 200 mg•kg-1 (M2) were conducted in a field at agricultural experiment station of Shandong Agricultural University in summer crop seasons of 2008 and 2009, using the typical varieties of edible sweet potato Beijing 553. There were three replicates for each treatment. The changes of starch accumulating characteristics and activities of enzymes involved in starch synthesis were determined by using periodic sampling during storage root development.【Result】Paclobutrazol significantly improved the activities of sucrose synthesis (SS) and adenosine diphosphorate glucose pyrophosphorylase (ADPGase), reduced the sucrose content in storage root. As a result, the starch accumulating rate and starch content were improved, and the starch production increased. Increase of the concentrations of paclobutrazol significantly increased the fresh weight per lump and fresh yield. Correlation analysis showed that ADPGase activity was positively correlated with starch content and starch accumulation rate and the correlation coefficients were 0.688 and 0.714, respectively (P<0.05). The correlation analysis also showed that SS activity was significantly positively correlated with starch content as well as the starch accumulating quantity and the correlated coefficients were 0.58 and 0.81 respectively (P<0.05).【Conclusion】The results suggested that spraying paclobutrazol increased yield of storage root, which resulted from the increased starch content and the improved starch accumulating rate. The higher activities of SS and ADPGase were found in resulting in higher yield of storage root in sweet potato.