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Table of Content

    15 August 2010, Volume 43 Issue 16
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Improving Resistance of Japonica Varieties Shengdao13 and Shengdao14 to Rice Stripe Virus Disease by Molecular Marker-Assisted Selection
    CHEN Feng,ZHANG Shi-yong,ZHU Wen-yin,ZENG Sheng-yuan,YANG Ya-chun,YUAN Shou-jiang,YANG Lian-qun
    Scientia Agricultura Sinica. 2010, 43(16):  3271-3279 .  doi:10.3864/j.issn.0578-1752.2010.16.001
    Abstract ( 885 )   PDF (335KB) ( 1126 )   Save
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    【Objective】 To develop new varieties and germplasms resistance to rice stripe disease (RSD), the molecular marker-assisted selection (MAS) strategy was used to improve the resistance of susceptible varieties Shengdao 13 and Shengdao14 .【Method】 Molecular markers closely linked to Stv-bi were used for MAS. Shengdao 13 and Shengdao14 were used as receptor parents, resistant variety Zhendao 88 and the high generation of disease-resistant rice strain Shengdao519 were used as the donor parents, applied self-improvement and backcross breeding methods, using MAS, combined with field resistance to RSD identification and agronomic traits analysis, Stv-bi (or allele gene) was successfully transferred to Shengdao13 and Shengdao14. 【Result】 A number of strains carrying the disease-resistance gene had been obtained through two inbred compound breeding populations, and six backcross stable lines carrying Stv-bi gene E01, E13, R107, etc. were obtained. 【Conclusion】 MAS can significantly improve the selection efficiency and accuracy in resistance breeding, MAS breeding system for stripe disease was established. The results have provided theories and materials for the future resistance breeding.

    Isolation and Expression Analysis of a Drought-Induced Gene ZmBTF3b in Maize (Zea mays L.)
    ZHANG Zhong-bao,ZHANG Deng-feng,LI Hui-yong,LIU Ying-hui,SHI Yun-su,SONG Yan-chun,WANG Tian-yu,LI Yu
    Scientia Agricultura Sinica. 2010, 43(16):  3280-3287 .  doi:10.3864/j.issn.0578-1752.2010.16.002
    Abstract ( 1214 )   PDF (822KB) ( 1283 )   Save
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    【Objective】 ZmBTF3b gene was cloned and used to analyze the expression profiles under abiotic stresses, providing a foundation for investigating mechanisms of molecular regulation in stress tolerance in maize. 【Method】 The promoter of ZmBTF3b was analyzed via bioinformatics. Transcriptional activation activity of ZmBTF3b was conducted using yeast one-hybrid system. Expression profiles of ZmBTF3b in different tissues of maize and in response to abiotic stresses were assayed via real-time quantitative PCR (qRT-PCR). 【Result】 The cloned ZmBTF3b gene encoded a putative transcription factor of 169 amino acids with a conserved NAC (nascent polypeptide-associated complex) domain. Transcriptional activity analysis showed that ZmBTF3b might be a functional transcription factor. Real-time quantitative PCR (qRT-PCR) analysis revealed that ZmBTF3b was highly expressed in silks, ears and immature embryos and was up-regulated under dehydration or PEG treatment while was down-regulated under cold, NaCl, ABA or SA. 【Conclusion】 ZmBTF3b might have the transcriptional activation activity and play an essential role in response to abiotic stresses.

    Identification and Genetic Analysis of Orchardgrass (Dactylis glomerata L.) Hybrids by SRAP Molecular Markers
    XIE Wen-gang,ZHANG Bao-yi,ZHANG Xin-quan,LIU Wei,CHEN Yong-xia,ZHOU He
    Scientia Agricultura Sinica. 2010, 43(16):  3288-3295 .  doi:10.3864/j.issn.0578-1752.2010.16.003
    Abstract ( 851 )   PDF (756KB) ( 1184 )   Save
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    【Objective】 Molecular markers were used to identify orchardgrass hybrids and analyze genetic diversity for forming orchardgrass hybrid identification system, detecting genetic information and conservation and utilization of orchardgrass germplasm resources. 【Method】 The sequence-related amplified polymorphism (SRAP) were assessed to analyze 140 orchardgrass hybrid individuals and their parental lines (01996, YA02-103) for detecting true or false hybrids according to polymorphism, and the genetic variation among hybrid individuals were detected using molecular markers and morphological markers. 【Result】 Sixty-four pairs of primers that generated clear, reproducible and polymorphic bands were selected from 192 pairs of SRAP primers with an average of 33.33%. 106 progenies with the special male bands compared with the female bands were identified from 140 progenies, which were therefore recognized as true hybrids. Sixteen primer pairs produced a total of 151 bands, of which 71 bands were polymorphic bands and the average polymorphic bands was four per primer pair. The average percentage of polymorphic bands was 47.24%. 【Conclusion】 These results suggested that SRAP technique has wide prospective in hybrid identification and diversity analysis of orchardgrass.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Relationship of PAR Interception of Canopy to Leaf Area and Yield in Rice
    LI Yan-da,TANG Liang,ZHANG Yu-ping,ZHU Xiang-cheng,CAO Wei-xing,ZHU Yan
    Scientia Agricultura Sinica. 2010, 43(16):  3296-3305 .  doi:10.3864/j.issn.0578-1752.2010.16.004
    Abstract ( 1271 )   PDF (633KB) ( 2325 )   Save
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    【Objective】 The amount of photosynthetically active radiation (PAR) interception and distribution within canopy affects crop photosynthesis and biomass production. The primary objective of this study was to elucidate the relationship of PAR interception of canopy to leaf area and yield of rice (Oryza sativa L.) under different nitrogen rates for different cultivar types.【Method】 Two field experiments were conducted at Nanjing in 2007 and 2008 rice growing seasons, involving two rice cultivars and three nitrogen rates. The amount of PAR interception and its distribution at different canopy heights, leaf area and yield of two cultivars were measured from jointing to maturity. 【Result】 The results indicated that the vertical distribution of upward cumulative leaf area index (LAI) followed a sigmoid curve, which could be described well with Logistic equation (R2>0.99). The maximum leaf area density in heading, 17 days after heading and maturity stages appeared at 0.53, 0.56 and 0.60 of relative canopy height, respectively. The relative leaf area density of upper and middle canopy increased with rice development progress, but the relative leaf area density of basal canopy decreased with rice development progress. The relationship between fraction of PAR interception (FIPAR) and downward cumulative LAI could be described with a negative exponential equation as FIPAR=α×(1-e-K×LAI) (R2>0.86). Diurnal variation of extinction coefficient (K) at noon was smaller than in the morning and afternoon, and the value of K decreased with rice development progress. The amount of PAR interception (AIPAR) showed multi-peaks distribution during rice development period, and the maximum peak appeared at 58-70 d after transplanting, around booting to heading stages. The AIPAR increased with increasing nitrogen application rate. Diurnal variation of AIPAR showed one peak distribution on the clear days, and the maximum AIPAR appeared at 11:00-13:00 in rice canopy. 【Conclusion】 The vertical distribution of leaf area affects PAR interception within rice canopy. The grain yield of rice was positively related to the PAR use efficiency, while PAR conversion efficiency first increased and then decreased with increasing PAR interception, thus it is benefit to have some penetration loss of PAR in rice canopy for high grain yield.

    HJ-1 Remotely Sensed Data and Sampling Method for Wheat Area Estimation
    ZHANG Jin-shui,SHEN Ke-jian,PAN Yao-zhong,LI Ling-ling,HOU Dong
    Scientia Agricultura Sinica. 2010, 43(16):  3306-3315 .  doi:10.3864/j.issn.0578-1752.2010.16.005
    Abstract ( 854 )   PDF (807KB) ( 974 )   Save
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    【Objective】 The method combing the advantages of remotely sensed data and sampling has showed the better actual value in practice for solving the problems about mis-registration error and regional difference. 【Method】 Taking Beijing as a study area, a combing method with multi-temporal HJ-1 remotely sensed data was used for wheat planting area estimation. Firstly, the multi-temporal remote sensing images were adopted for the wheat area, then integrating the cultivated parcel for building sample populations. Secondly, with wheat area in the parcel as the stratified flag, the stratified random sample method was applied. Thirdly, after ground survey for every sample, the overall regional wheat area was derived. The results showed that MAE and bias was 0.17 and -0.05 respectively. 【Result】 The sample method increased by 5% for regional wheat planting area, which corrected some remote sensing errors. 【Conclusion】 It was concluded that the method can guarantee the regional and parcel scale accuracy, and can be broadly used in crop area estimation.

    Biomass Yield and Quality of Three Kinds of Bioenergy Grasses in Beijing of China

    FAN Xi-feng,HOU Xin-cun,ZUO Hai-tao,WU Ju-ying,DUAN Liu-sheng
    Scientia Agricultura Sinica. 2010, 43(16):  3316-3322 .  doi:10.3864/j.issn.0578-1752.2010.16.006
    Abstract ( 867 )   PDF (327KB) ( 1553 )   Save
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    【Objective】 The objective of this experiment was to evaluate the yield and biomass characteristics of three herbaceous plants, namely switchgrass (Panicum virgatum L.), silverreed (Triarrhena sacchariflora (Maxim.) Nakai) and giantreed (Arundo donax L.), in Beijing of China for their rational utilization as bioenergy feedstocks. 【Method】 Field trials were conducted at experimental station of Beijing Research & Development Center for Grasses and Environment from 2006 to 2009 and the biomass yield and quality parameters were investigated by field observations and laboratory analysis. 【Result】 The dry biomass yield was 28.33, 29.67, and 34.46 t?hm-2 for switchgrass, silver reed, and giant reed, respectively. Among the three grasses, switchgrass and silverreed would be perfect for cellulosic ethanol conversion due to higher biomass content of cellulose and hemicellulose and lower lignin content, for direct combustion, co-combustion with coal, pyrolysis and gasification due to higher calorific value, ash melting temperature, and contents of C and H elements and lower content of N, ash, and moisture, but disadvantaged for methane fermentation because of higher ratio of C to N. Giantreed had higher biomass content of water, lignin, and ash than those of switchgrass and silverreed and would be disadvantaged for transport, storage, ethanol conversion, and combustion. 【Conclusion】Switchgrass, silverreed, and giantreed, were appropriate for extended planting in Beijing and surrounding areas as excellent herbaceous energy plants. But reasonable species choice should be employed according to the conversion approach and the growth characteristics, productivity levels, and biomass quality characteristics of the three grasses.

    PLANT PROTECTION
    Cloning and Expression Analysis of the PVY Resistance-Related Gene NtPsaN in Tobacoo (Nicotiana tabacum)
    ZHOU Jia,LI Feng-xia,CHEN Shuai,LUO Cheng-gang,LIU Guan-shan,JIANG Cai-hong,YANG Ai-guo,SU Zhen-gang,WANG Yuan-ying
    Scientia Agricultura Sinica. 2010, 43(16):  3323-3330 .  doi:10.3864/j.issn.0578-1752.2010.16.007
    Abstract ( 1023 )   PDF (642KB) ( 1280 )   Save
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    【Objective】 The common tobacco (Nicotiana tabacum) variety VAM was highly resistant to PVY. A PVY resistance-related gene NtPsaN was cloned from VAM. The sequence features, evolutionary relationships and expression characteristics of NtPsaN were analyzed and the molecular mechanism of tobacco resistance response was studied, and thus provide a theoretical foundation for PVY-resistant tobacco breeding. 【Method】 A virus resistance-related gene fragment was screened through Suppression Subtractive Hybridization (SSH) and cDNA chip technologies, the full-length cDNA sequence was cloned through RACE, the conserved domains and sequence features of the gene were analyzed by bioinformatic methods, a neighbor-joining phylogenetic tree was constructed using the MEGA4.0 program and its relative expression characters were explored by real-time PCR. 【Result】 The tobacco resistance-related gene, named NtPsaN, was cloned. The gene possesses conserved domains of PsaN superfamily. The open reading frame (ORF) of the full length cDNA was 507 bp in length, encoding 168 amino acids. The PsaN subunit phylogenetic tree was constructed. The results of real-time PCR showed that the expression of NtPsaN was obviously up-regulated in leaves infected with PVY. 【Conclusion】 A PVY resistance-related gene NtPsaN was cloned from tobacco, and it was induced by PVY infection. The NtPsaN gene may play a critical role in the PVY-resistance of tobacco.

    Cloning and Analysis of an Aspartic Protease Gene Ntasp Induced by PVY in Tobacco (Nicotiana tabacum)
    CHEN Shuai,LIU Guan-shan,ZHOU Jia,YANG Ai-guo,WANG Yuan-ying,SUN Yu-he
    Scientia Agricultura Sinica. 2010, 43(16):  3331-3339 .  doi:10.3864/j.issn.0578-1752.2010.16.008
    Abstract ( 862 )   PDF (884KB) ( 950 )   Save
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    【Objective】 The objective of this study was to screen and clone PVY infected resistance-related genes, and to analyze their relative expression quantity, explore the molecular mechanism of PVY resistance and provide a theoretical foundation for PVY-resistance tobacco (Nicotiana tabacum) breeding. 【Method】 A middle gene fragment of up-regulated expression (Ratio>2) was screened from the tobacco leaf suppression subtractive hybridization (SSH) library induced by PVY using SSH and cDNA chip technologies, its full-length cDNA sequence was cloned through rapid amplification of cDNA ends (RACE), and its relative expression quantity in different infection times was analyzed by real-time quantitative PCR. 【Result】 A middle gene fragment of 595 bp was screened from tobacco leaves induced by PVY, and a full-length tobacco aspartic protease cDNA of 1 770 bp was cloned by RACE and named Ntasp (GenBank accession no.GU144571), which encoded a protein of 506 amino acids. Sequence alignment indicated that Ntasp revealed a high degree of similarity with other members of plant aspartic proteases and had a typical domain characteristic. The results of real-time quantitative PCR showed that expressions of Ntasp transcripts were up-regulated in tobacco leaves infected by PVY in early stage. 【Conclusion】 A tobacco aspartic protease gene Ntasp was cloned and induced by PVY infection.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Impact of Long-Term Fertilization on the Microbial Biomass Carbon and Soil Microbial Communities in Paddy Red Soil
    BU Hong-zhen,WANG Li-hong,YOU Jin-cheng,XIAO Xiao-ping,YANG Guang-li,HU Yue-gao,ZENG Zhao-hai
    Scientia Agricultura Sinica. 2010, 43(16):  3340-3347 .  doi:10.3864/j.issn.0578-1752.2010.16.009
    Abstract ( 832 )   PDF (347KB) ( 1594 )   Save
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    【Objective】 Soil microorganisms are involved in various biochemical processes and play a vital role in maintaining soil fertility and plant yields. The impacts of different long-term fertilization treatments on soil microbial community structure were studied to clarify the relationship between them. 【Method】 Soil samples were collected from long-term fertilization experimental plots receiving no fertilization, chemical fertilization, chemical fertilization as normally applied by local farmers, harvested residue return, 60% organic manure plus chemical fertilization and 30% organic manure plus chemical fertilizer. Total microbial biomass carbon was determined by chloroform- fumigation-extraction. Phospholipid fatty acid (PLFA) profiles for each treatment were used to characterize the microbial community structure and were analyzed by principle component analysis. 【Result】 Compared with the unfertilized control, all fertilization treatments showed significant increases in total microbial biomass carbon and PLFAs. The iC15:0 fatty acids significantly increased under long-term application of chemical fertilizer but decreased under both organic manure and residue return fertilization. In contrast, under organic manure and residue return, the aC15:0 fatty acids increased. Principal component analyses of the 21 PLFAs detected in the six treatments showed that 87.6 % of the total variance of the individual PFLAs could be explained by two principal components. All unsaturated and cyclopropyl PLFAs except C12:0 and C15:0 were highly weighted on the first principal component. The first and the second principal components could also explain 89.8% of the total variance of fertilization treatment. There was a very little different between the chemical fertilizer treatment and the treatment receiving chemical fertilizer as typically applied by local farmers. There was also almost no difference between the treatment that received residue return and the treatment that received 60% organic manure plus chemical fertilizer. 【Conclusion】 Long-term fertilization can significantly impact soil microbial biomass carbon and influence microbial community structure. PLFAs reflect soil microbial community structure and can be used to detect shifts in response to fertilization.

    Soil Desiccation of Alfalfa Fields and Effects of Alfalfa-Grain Crop Rotation on Soil Water Restoration of Desiccated Alfalfa Fields in Semi-Humid Areas of the Loess Plateau
    FANG Xin-yu,LI Jun,WANG Xue-chun,REN Jing-jing
    Scientia Agricultura Sinica. 2010, 43(16):  3348-3356 .  doi:10.3864/j.issn.0578-1752.2010.16.010
    Abstract ( 987 )   PDF (337KB) ( 835 )   Save
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    【Objective】 Soil desiccation in alfalfa fields and the effects of alfalfa-grain crop rotation on soil water restoration of desiccated alfalfa fields in semi-humid areas of the Loess Plateau were studied. 【Method】 Soil moisture of alfalfa fields with different alfalfa-growing years and various alfalfa-grain crop rotation fields were observed at Zhenyuan Station of Gansu province, and the characteristics of desiccated soil layers of alfalfa fields and the effects of soil water restoration by alfalfa-grain rotation were analyzed.【Result】 The average soil moisture in the 0-1 000 cm soil layers of 15-28-year alfalfa fields was 10.20%, soil desiccation rate was 34.2 mm per year and the maximum distribution depth of desiccated soil layer was over 1 400 cm. Soil moisture in desiccated soil layer of alfalfa fields could be restored gradually after alfalfa plowed out and rotated with grain crops for 3-25 years, the average thickness of moisture restored soil layers was 583 cm, and the average rate of soil water resumption was 77.3 mm per year, and the average soil water restoration index (SWRI) reached 83.3%. To restore soil water to local soil stable moisture (15.5%) in 15 alfalfa-growing years field by alfalfa-grain crops rotation at least 8 years are needed. 【Conclusion】 The suitable alfalfa-grain crops rotation pattern for soil water restoration is “10 year alfalfa rotated with grain crops for at least 8 years”.

    Effects of Soil Tillage on CH4 Emission During Paddy Season in a Rice-Wheat Double Cropping System
    ZHANG Yue-fang,ZHENG Jian-chu,CHEN Liu-gen,WANG Zi-chen,ZHU Pu-ping,SHENG Jing,WANG Ya-lei
    Scientia Agricultura Sinica. 2010, 43(16):  3357-3366 .  doi:10.3864/j.issn.0578-1752.2010.16.011
    Abstract ( 1001 )   PDF (384KB) ( 1029 )   Save
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    【Objective】 CH4 emission during paddy season under different soil tillages in a rice-wheat double cropping system were investigated for providing scientific instructions to develop rational measures of reducing greenhouse gas emission in the lower reaches of the Yangtze River. 【Method】 A split plot experiment was conducted to study the effects of soil tillage before wheat sowing and soil tillage before rice transplanting on CH4 emission during paddy season by using the method of static chamber-gas chromatographic techniques. Soil tillage before wheat sowing was designed as no tillage before wheat sowing (NTW), rotary tillage before wheat sowing (RTW) and plow tillage before wheat sowing (PTW). Soil tillage before rice transplanting was designed as rotary tillage before rice transplanting (RTR) and plow tillage before rice transplanting (PTR) for each soil tillage before wheat sowing just before rice transplanting. 【Result】 The seasonal variation characteristics of CH4 emission under different soil tillages in a rice-wheat double cropping system were defined, and represented an increasing at the beginning and declining later on. CH4 accumulative emissions from transplanting stage to critical stage of productive tillering accounted for 64.73%-86.84% of the total emission during paddy season. Under NTW, average CH4 emission during paddy season was 53% and 24% higher than under RTW and PTW, respectively, and they were significantly different. Average CH4 emission during paddy season under PTR was significantly higher than under RTR. Total CH4 emissions and total CH4 emissions per unit area yield under different soil tillage followed the same order: NTW+PTR>NTW+RTR>PTW+PTR>PTW+RTR>RTW+PTR>RTW+RTR. Correlation analysis showed that CH4 emission flux under different soil tillages was negatively related to soil Eh during paddy season, and a significantly positive correlation between total CH4 emissions and soil organic matter content at 0-5 cm soil layer was also found.【Conclusion】 Total CH4 emissions during paddy season were significantly influenced by soil tillage before wheat sowing and soil tillage before rice transplanting. Annual rotary tillage is one of the effective measures to reduce total CH4 emissions during paddy season from rice-wheat double cropping system in the lower reaches of the Yangtze River.

    HORTICULTURE
    Study on Relationship Among Microorganism, Enzymes’ Activity in Rhizosphere Soil and Root Rot Resistance of Grafted Pepper
    JIANG Fei,LIU Ye-xia,AI Xi-zhen,ZHENG Nan,WANG Hong-tao
    Scientia Agricultura Sinica. 2010, 43(16):  3367-3374 .  doi:10.3864/j.issn.0578-1752.2010.16.012
    Abstract ( 1186 )   PDF (379KB) ( 1394 )   Save
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    【Objective】 The purpose of this study is to elucidate the mechanism of root rot resistance in grafted pepper (Capsicum annuum L.), and offer a theoretical basis for regulating root environment and alleviating root rot damage. 【Method】 Changes of disease incidence and disease index of root rot in rootstock (‘Weishi’), grafted (‘Xinfeng 2’ scion grafted onto ‘Weishi’) and own-root plants (‘Xinfeng 2’, control) of pepper, root absorption area, root respiration intensity, population of microoganism and enzymes’ activity in rhizosphere soil were investigated by manual inoculation. 【Result】 The results showed that the root absorption area, actinomyces population and ratio, activities of dehydrogenase, peroxidase and catalase in rhizosphere soil of rootstock and grafted peppers increased significantly compared with those of control before inoculation. After been inoculated, grafted plants showed remarkable lower disease incidence and disease index than control plants, but higher than rootstock plants. Rootstock and grafted peppers had significant higher root absorption area and active absorption area, quantities of bacterium, fungus and actinomyces, actinomyces ratio, and soil enzymes’ activity, in comparison with control. The root respiration rate was lower in rootstock and grafted plants than in control plants at earlier stage, but just the opposite at later stage. 【Conclusion】 The observed increase in root absorption area, quantity of microoganism, ratio of actinomyces, and enzymes’ activity in rhizosphere soil of grafted peppers are important reasons of enhancement of root rot resistance.

    Function Analysis of MbNramp1 Gene from Malus baccata (L.) Borkh Through Yeast Complementation Experiments
    XIAO Hai-hua,YIN Li-ping,HAN Zhen-hai
    Scientia Agricultura Sinica. 2010, 43(16):  3375-3380 .  doi:10.3864/j.issn.0578-1752.2010.16.013
    Abstract ( 853 )   PDF (476KB) ( 1354 )   Save
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    【Objective】 The objective of the study is to investigate the function of MbNramp1 gene. 【Method】 Heterogenous complementation experiments were employed to confirm the iron- transporting ability of MbNramp1,and study subcellular localization of the MbNRAMP1 protein 【Result】 MbNramp1 was expressed in yeast mutant DDY4, which could restore growth in the media without iron. At tested concentrations of BPDS, the yeast cells DDY4 mutant after MbNramp1 being transformed grew better than mutant strain transformed with empty vector. When the concentration of BPDS in the media reached 15 μmol?L-1, the growth of yeast cells with MbNramp1 was comparable with that of wild type. Although the growth of MbNramp1 transformed cells in the media with 30 μmol?L-1 BPDS was slower than those in the media with low concentration of BPDS, they were significantly faster that DDY4 mutant transformed with empty vector. In addition, wild type grew slower with 30 μmol?L-1 BPDS. The result of subcellular localization showed that MbNRAMP1 proteins were mainly positioned in the part of plasma membrane not in the whole membrane. 【Conclusion】 Initial results suggested that MbNRAMP1 protein has the ability of transferring Fe and also help yeast mutant to grow. In addition, MbNRAMP1 proteins were mainly positioned in the part of plasma membrane

    Organic Acids in Apple Trees and Their Effects on Zinc Uptake and Distribution Under Zinc Deficiency
    LIU Di,LIU Ai-hong,WANG Jin-hua,ZHANG Yuan-zhen,WANG Yan-an,ZHANG Fu-suo,SHU Huai-rui
    Scientia Agricultura Sinica. 2010, 43(16):  3381-3391 .  doi:10.3864/j.issn.0578-1752.2010.16.014
    Abstract ( 807 )   PDF (646KB) ( 1033 )   Save
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    【Objective】 To explore the functional mechanism of the effect of organic acids on Zn uptake and distribution, organic acids in apple cultivar ‘Fuji’ in the field, two-year old ‘Fuji’ and rootstock Malus hupehensis (Pamp.) Rehd. seedlings were detected, and their effects on Zn uptake and distribution under Zn deficiency was studied. 【Method】 ‘Fuji’ was sampled at bud, florescence and physiological fruit dropping stages in the field, two-year old ‘Fuji’ in sand culture, Malus hupehensis (Pamp.) Rehd. seedlings were grown hydroponically and two treatments (pZn2+ 11.3, pZn2+ 10.7) were conducted for 1 d and 20 d before determination of organic acids. 【Result】 In R0-1.5, R1.5-3, R3-5 and R5-15, Zn and organic acids concentrations were mostly significantly higher in rosette tree than that in normal tree at bud stage while mostly lower at florescence stage, however, at physiological fruit dropping stage, Zn concentration of root was higher in rosette tree than that in normal tree, organic acids concentrations decreased in R0-1.5 and R1.5-3 while mostly increased in R3-5 and R5-15 of rosette tree. The rhythm of Zn absorption and organic acid metabolism were changed, organic acid levels were highly related to the total Zn levels in root of ‘Fuji’ in field and of hydroponical Malus hupehensis (Pamp.) Rehd. The organic acids concentrations in absorbing roots of two-year old ‘Fuji’ rose under Zn deficiency. The hydroponical Malus hupehensis (Pamp.) Rehd experiment showed that compared with pZn2+ 10.7, pZn2+ 11.3 treatment for 1 d, Zn concentration in root decreased significantly, while oxalic acid and malic acid concentrations in stem and root increased by 1.09-1.35 fold, the distribution proportion of root organic acid elevated, Zn uptake rate significantly increased. While treatment for 20 d, Zn and organic acids concentrations, organic acid distribution proportion of leaf significantly decreased.【Conclusion】 The rhythm of Zn absorption and utilization and organic acid metabolism and transportation were changed by Zn deficiency, which promoted organic acids transport from shoot to root. Organic acids play an important role in promoting Zn uptake and distribution.

    Cloning of Mitochondrial F1-ATPase Beta Subunit Gene from Embryogenic Callus and Its Expression Analysis by qRT-PCR During Somatic Embryogenesis in Longan

    LAI Cheng-chun,LAI Zhong-xiong,FANG Zhi-zhen,LIN Yu-ling,JIANG Shun-ri
    Scientia Agricultura Sinica. 2010, 43(16):  3392-3401 .  doi:10.3864/j.issn.0578-1752.2010.16.015
    Abstract ( 915 )   PDF (523KB) ( 853 )   Save
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    【Objective】 The mitochondrial F1-ATPase beta subunit gene from embryogenic callus was cloned and its expression during somatic embryogenesis in longan (Dimocarpus longan Lour.) was analyzed in this experiment. 【Method】 The complete cDNA sequence of the mitochondrial F1-ATPase beta subunit gene of longan embryogenic callus was obtained by reverse transcription polymerase chain reaction (RT-PCR) with rapid amplification of cDNA ends (RACE). And the mRNA transcription level of this gene was surveyed by real-time reverse transcription PCR (qRT-PCR) during somatic embryogenesis in longan. 【Result】 The full-length cDNA sequence of longan embryogenic callus mitochondrial F1-ATPase beta subunit gene was 2 099 bp (GenBank accession number FJ222749). It contained a 1 677-nucleotides-long open reading frame (ORF) which encoded a protein of 588 amino acid residues. Homology searches with the deduced amino acid residues indicated that longan mitochondrial F1-ATPase beta subunit gene had a high similarity to other plant mitochondrial F1-ATPase beta subunit gene. According to the 28 mitochondrial F1-ATPase beta subunit gene sequences of animals and plants, the phylogenetic tree showed that it indeed reflected the evolution of animals and plants. It had a single branch because longan mitochondrial F1-ATPase beta subunit gene was firstly cloned. The results showed that the mRNA transcription levels of longan mitochondrial F1-ATPase beta subunit gene were different at the different stages during longan somatic embryogenesis. The highest level occurred at the globular embryo stage and the lowest at the torpedo embryo stage. 【Conclusion】 Longan mitochondrial F1-ATPase beta subunit gene had a high similarity to other plant mitochondrial F1-ATPase beta subunit gene. The highest mRNA transcription level of longan mitochondrial F1-ATPase beta subunit gene occurred at the globular embryo stage.

    STORAGE·FRESH-KEEPING·PROCESSING

    Effects of CaCl2 and AsA Treatments on Quality and Browning in Fresh-Cut Apple

    YANG Wei,LIU Jing,Lü Chun-jing,WANG Jie,ZHANG Su-min
    Scientia Agricultura Sinica. 2010, 43(16):  3402-3410 .  doi:10.3864/j.issn.0578-1752.2010.16.016
    Abstract ( 1015 )   PDF (348KB) ( 1504 )   Save
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    【Objective】 The changes of quality and browning of the fresh-cut apples, the physiological roles of exogenous ascorbic acid (AsA) and calcium chloride (CaCl2) were studied for screening of an ideal preservative. 【Method】 Ripening ‘Fuji’ apple fruits were used as materials and peeled, cored, and then cut into slices about 1mm; after soaked for 5 min in distilled water, 1.0% AsA and 1.0% CaCl2 aqueous solution, wiped the surface water, then put in culture vessel under 25 ℃, and at 0, 0.25, 0.5, 1, 2, 4, 6, 8, 10, 12 h, the browning index and the rate of weigh loss were surveyed, the contents of total soluble sugars, total titratable acids, ascorbic acid (AsA), total phenols and malondialdehyde (MDA) were analyzed, and the polyphenol oxidase (PPO) and peroxidase (POD) activities were detected. 【Result】 The quality and physiological characteristics of the fruit slices soaked in distilled water were changed dramatically, flesh browning was rapid and serious, water and other volatile material released largely, the content of total soluble sugars, total titratable acids, AsA and total polyphenols decreased, the ratio of sugar and acid increased, the activities of PPO and POD rose rapidly. The content of MDA increased first and then decreased. 1.0% AsA and 1.0% CaCl2 treatment could effectively delay the process of flesh browning, reduced browning degree, inhibited decreasing of water, volatile substances, total soluble sugars, total titratable acids, ascorbic acid and total polyphenols, delayed sugar/acid ratio increase, as well as PPO, POD activities in the peak arrival, the content of MDA increased. In general, the treatment of 1.0% CaCl2 was better. In addition, a quality evaluation system of fresh-cut product was initially established. Varieties with high content of soluble solids, total soluble sugar, total titratable acids and phenolic substances, low volatile matter loss rate, and browning-resistance are good apple varieties for fresh-cut apples processing.【Conclusion】 1.0% CaCl2 is a better preservative reagent and browning inhibitor in fresh-cut apples.

    Plasma Membrane H+-ATPase Involved in Mechanical Wounding-Induced Defense Response in Pea Leaves
    LIU Yan,HUANG Wei-dong,CHEN Gui-lin,YUE Xin
    Scientia Agricultura Sinica. 2010, 43(16):  3411-3417 .  doi:10.3864/j.issn.0578-1752.2010.16.017
    Abstract ( 1015 )   PDF (419KB) ( 673 )   Save
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    【Objective】 Effects of plasma membrane H+-ATPase (PM H+-ATPase) on defense response to mechanical wounding were investigated. 【Method】 Using pea (Pisum sativum L.)seedlings as materials, treated by mechanical wounding, and using methods of enzymes activity measures, western blot detection and electron microscope cytochemistry, the effects of mechanical wounding on PM H+-ATPase activities were observed. Furthermore, the relationship between PM H+-ATPase and accumulation of active oxygen in inducing wounding defense response was studied. 【Result】 Mechanical wounding induced the increasing of PM H+-ATPase activity, but did not change its content; suppressed PM H+-ATPase activity, inhibited H2O2 accumulation which has been induced by wounding and also inhibited the phenylalanine ammonia lyase activity's increasing, while application of active oxygen scavengers had no effect on the wounding -induced PM H+-ATPase activity increasing. 【Conclusion】It is thus suggested that PM H+-ATPase participated in signal transduction of defense response to mechanical wounding, which acted on the upstream of reactive oxygen signal and induced the formation of defense response on the downstream by protein phosphorylation regulation.

    ANIMAL SCIENCE·RESOURCE INSECT
    Effect of Nitrate Addition Level on in vitro Ruminal Fermentation Characteristics and Microbial Efficiency
    DAI Jun-fang,MENG Qing-xiang,ZHOU Zhen-ming,CUI Zhen-liang,REN Li-ping
    Scientia Agricultura Sinica. 2010, 43(16):  3418-3424 .  doi:10.3864/j.issn.0578-1752.2010.16.018
    Abstract ( 883 )   PDF (310KB) ( 965 )   Save
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    【Objective】 The study was conducted to evaluate the effect of dietary nitrate N addition level on the rumen fermentation and microbial efficiency. 【Method】 The experimental diets were formulated with six dietary nitrate N addition levels including 0, 6%, 12%, 18%, 24% and 30%, respectively. The fermentation in vitro was carried out in dual-flow continuous culture system lasting for 144 h. 【Result】 As dietary nitrate N addition level increased, ruminal pH and total VFA of fermenter contents were unchanged (P>0.05), while the molar percentages of propionic acid (Q, P<0.01) increased, and acetic acid (Q, P<0.01) and ammonia concentration decreased (Q, P<0.01). At the same time, there were quadratic increases (Q, P<0.01) in the digestibility of DM, OM and CP. Microbial efficiency (MOEEF) increased quadratically (Q, P<0.01) as dietary nitrate N addition level increased.【Conclusion】 Theses results indicated that dietary nitrate N could be used by ruminal microorganisms. As dietary nitrate N addition level increased, the ruminal fermentation parameters, digestibilities and MOEEF changed in varying degrees. Besides, ruminal MOEEF and fermentability were greater at the dietary nitrate N addition range of 12% to 18%.

    Polymorphic Analysis on Microsatellite Marker and Its Growth Index Association of Bashbay Sheep
    ANIWASHI J K,HAN Ye-dong,LI Qi-fa,YIDULA P N E,XIE Zhuang
    Scientia Agricultura Sinica. 2010, 43(16):  3425-3432 .  doi:10.3864/j.issn.0578-1752.2010.16.019
    Abstract ( 812 )   PDF (405KB) ( 886 )   Save
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    【Objective】 Taking Bashbay sheep as research material, this paper aimed at seeking the microsatellite marker which influenced the growth property of Bashbay sheep through analysis on the genotype of microsatellite DNA locus and growth property, and thus providing a basis for the molecular marker assisted selection. 【Method】 Inquiried about the microsatellite locus of sheep and goat from GenBank database, 10 microsatellite markers that meet the requirements of the microsatellite standard were selected, 189 individual genome DNA of sheep were detected, and the correlated effects of microsatellite locus and growth indexes were analyzed by adopting least-squares method. 【Result】 There were 110 allelic genes were detected by 10 microsatellite markers, with an average of 11 allelic genes for each, the average polymorphic information content was 0.7914, the average heterozygosity was 0.8149. There were 8 of 10 microsatellite locus had significant and very significant relation with 5 growth indexes including weight and size of Bashbay sheep, especially the significant correlation of gene locus BM415 and weight and size (P<0.01). Multiple comparative results showed that 8 microsatellite locus had genotype advantage.【Conclusion】 Significant effect was found in the growh indexes of Bashbay sheep group, and the higher microsatellite locus offered valuable genetic markers for developing the molecular marker assisted breeding of growth indexes of Bashbay sheep.

    Effect of De-oiled DDGS on Production Performance, Egg Quality and Serum Biochemical Indexes in Laying Hens
    LI Yu,ZHANG Jun-min,MENG Yan-li,ZHAO Qing-yu,DENG Li-kang
    Scientia Agricultura Sinica. 2010, 43(16):  3433-3439 .  doi:10.3864/j.issn.0578-1752.2010.16.020
    Abstract ( 1198 )   PDF (305KB) ( 1063 )   Save
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    【Objective】 An experiment was conducted to study the effects of dietary content of de-oiled distillers dried grains with solubles (DDGS) on production performance, egg quality and serum biochemical indexes in laying hens. 【Method】Nine hundred and sixty Hy-line brown laying hens of 67 weeks old were randomly devided into 4 groups, each of which was replicated four times with sixty hens per replicate. The control group was fed with basal diet, and test I,test II and III groups were fed with diet containing 12%, 18% and 24% DDGS, respectively, for 8 weeks. 【Result】 In the whole experimental period, no significant differences (P>0.05) in feed intake, egg shape index, and eggshell thickness were observed among the four groups. However, the test groups had lower egg weights and deeper yolk color than control group (P<0.05). Compared with control group, there were no significant differences in feed-egg ratio, laying rate and Haugh unit in test I and test II (P>0.05), but test III had higher feed-egg ratio and Haugh unit,and lower laying rate (P<0.05). No significant differences (P>0.05) in serum biochemical indexes (Ca, P, TG, HDL and BUN) were observed among the four dietary treatments groups. 【Conclusion】 If dietary content of DDGS is less than 18%, there is no negative effects on feed-egg ratio, laying rate,egg quality and serum biochemical indexes in laying hens,but egg weight will be decreased. However, the feed-egg ratio will be increased and laying rate and egg weight will be decreased when the contens is up to 24%.

    VETERINARY SCIENCE
    Effects of Glucocorticoid and Tryptophan Perfusion on Hypothalamic Metabolites of Broilers
    KONG Xue,SONG Zhi-gang,JIAO Hong-chao,LIN Hai
    Scientia Agricultura Sinica. 2010, 43(16):  3440-3446 .  doi:10.3864/j.issn.0578-1752.2010.16.021
    Abstract ( 835 )   PDF (424KB) ( 958 )   Save
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    【Objective】 The effects of glucocorticoid and tryptophan on the hypothalamic metabolites concentration of broiler chickens were investigated by nuclear magnetic resonance (NMR) in the present study. The aim of the present study was to provide evidences for the understanding of hypothalamic regulation mechanism on appetite and expenditure at stress status. 【Method】 Thirty-two 10-day-old chickens were divided into 4 groups randomly, dexamethasone (DEX, 2.6 mg?kg-1 BW) was injected subcutaneously and tryptophan (500 mg?kg-1 BW) was oral administrated after 3 h fasting, while the control chickens were subjected to saline administration, respectively. Blood and hypothalamus were obtained and hypothalamus extraction was used for NMR analysis. 【Result】 The 1H NMR spectra showed that the relative concentrations of lactate, glucose, ATP, myo-inositol and 3-Hydroxybutyrate were changed by DEX treatment, while tryptophan signal was not significantly altered. 【Conclusion】 The result indicates that NMR could detect the changes in hypothalamic metabolism under stress status. The result suggests that the signals of energy substrates such as ATP and glucose may play a role in the central regulation of appetite of chickens.

    Detection of Plasmid-Mediated Quinolone Resistance in Clinical Isolates of Escherichia coli from Pet Animals
    ZHU Heng-qian,LIAO Xiao-ping,CHEN Chao-xi,WANG Xiu-mei,SUN Jian,SUN ying,LI Liang,ZHANG Mei-jun,LIU Ya-hong
    Scientia Agricultura Sinica. 2010, 43(16):  3447-3454 .  doi:10.3864/j.issn.0578-1752.2010.16.022
    Abstract ( 785 )   PDF (575KB) ( 797 )   Save
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    【Objective】 The plasmid-mediated quinolone resistance PMQR genes of qnr, qepA and aac (6′)-Ib-cr in clinical isolates of Escherichia coli from pet animals (dogs and cats) in Guangzhou city were detected. 【Method】 Susceptibility of 164 strains to 15 antimicrobial agents were determined by agar disc dilution method. All of the 164 strains were screened for the qnr, qepA, aac (6′)-Ib-cr genes by PCR. Genetic fingerprints of the clinical isolates were established and analyzed. 【Results】 The isolates showed a high rate of resistance to antimicrobial agents widely used in veterinary clinical medicine and most of the them showed multiple-resistance(5 to 14 drugs). Three of these 164 specimens were detected containing qnrA, qnrB and qnrS genes, and two of which also posses aac(6′)-Ib-cr gene. Fingerprints could be amplifled from all the positive strains, and strains with more than two PMQR genes were distributed mainly in the B and C gene types. 【Conclusion】 Escherichia coli isolated from pet animals in Guangzhou city showed a high level multiple-resistance to widely used drugs. The result of the research showed a more widespread dissemination of resistance to quinolones in pet animal veterinary clinic of Guangzhou city, and the ability to generate PMQR gene of isolates is closely to their gene-type.

    Construction of VP1 Recombinant Lentivirus Vector and Establishment of BHK-21 Cell Lines Stably Expressing VP1 Gene of FMDV
    DAI Wen-jun,WANG Hong-mei,LIU Xiao,GAO Yun-dong,YU Li,WANG Li-qun,ZHONG Ji-feng,HE Hong-bin
    Scientia Agricultura Sinica. 2010, 43(16):  3455-3460 .  doi:10.3864/j.issn.0578-1752.2010.16.023
    Abstract ( 1114 )   PDF (368KB) ( 714 )   Save
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    【Objective】 To construct the lentivirus vector containing VP1 gene and to establish the cell line with stable expression of VP1 gene of foot-and-mouth disease virus (FMDV). 【Method】 The full-length VP1 gene was amplified by RT-PCR, and VP1 gene was cloned into FG9 vector, then recombinant vector was confirmed by restricting enzyme digestion and DNA sequence. The recombinant plasmid was transfected into BHK-21 cells through LipofectamineTM 2000, and the GFP positive cells were screened via FACS after 96h. The expressions of VP1 gene were confirmed by Western-blot. 【Result】 FG9-VP1 was constructed successfully and the VP1 gene could be stably expressed in BHK-21 cel1s. 【Conclusion】 The recombinant lentivirus vector containing VP1 gene was cloned successfully and the stable cell line expressing the VP1 gene was established.

    RESEARCH NOTES

    Research on Aluminum Tolerance of Citrate Synthase Transgenic Alfalfa

    GAN Zhi-cai,CHEN Dong-ying,ZHANG Li,YU Yong-xiong
    Scientia Agricultura Sinica. 2010, 43(16):  3461-3466 .  doi:10.3864/j.issn.0578-1752.2010.16.024
    Abstract ( 890 )   PDF (249KB) ( 796 )   Save
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    【Objective】 An experiment on root tip tolerance of non-transgenic control (CK) and citrate synthase (CS) transgenic plants from Medicago sativa L. cv. Yumu No.1 was conducted to find out the suitable conditions for Al-tolerance and to examine the Al-tolerance of the transgenic plants relative to the control plants. 【Method】 Using CS transgenic alfalfa No.4 and the control plant as materials, five AlCl3 concentration gradients were designed which were 0, 5, 15, 30, and 50 μmol?L-1, each with three replicates, the root tip elongation in the first three days was measured. 【Result】 The root tip elongation of transgenic and control plants on the first day was significantly higher than on the second and the third day in all five AlCl3 concentrations. The root tip elongation of transgenic plants at 15, 30, and 50 μmol?L-1 AlCl3 concentrations on second day was significantly higher than on the third day, while the root tip of control plant almost stopped elongating on the second and third day. With increased Al concentrations, the root tip elongation was inhibited seriously. The root tip of control plants in 15 μmol?L-1 Al treatment could elongate normally on the first day, while the elongation was seriously suppressed even if in 5 μmol?L-1 Al treatment on second day, indicating that the Al concentration the control plants could be tolerant of was less than 15 μmol?L-1. When the AlCl3 concentration increased to 30 μmol?L-1, the root tip elongation of transgenic plants was significantly suppressed (P<0.05). 【Conclusion】 Al-tolerance conditions of transgenic alfalfa in Medicago sativa L. cv. Yumu No.1 were 15-30 μmol?L-1 AlCl3 concentrations for 2 days. Compared to the control, the transgenic alfalfa showed a significant resistance to Al.

    Multi-Residues Determination of Abamectin, Emamectin Benzoate and Ivermectin in Vegetables by HPLC-FD
    ZHAO Li,XIE Xian-chuan,ZHAN Xiu-ping
    Scientia Agricultura Sinica. 2010, 43(16):  3467-3472 .  doi:10.3864/j.issn.0578-1752.2010.16.025
    Abstract ( 966 )   PDF (257KB) ( 1287 )   Save
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    【Objective】 The aim of the study is to build a sensitive high-performance liquid chromatography with fluorescence detection (HPLC-FD) method for determinating multi-residues of abamectin(ABA), emamectin benzoate (EMA) and ivermectin (IVM) in vegetables. 【Method】 After being extracted by acetonitrile, the avermectins residues in vegetables were directly derivatised by N-methylimidazole (N-NMIM) and trifluoroacetic anhydride (TFAA) and then analyzed on HPLC-FLD using gradient elution program with acetonitrile/methanol/water. 【Result】 Results showed that the developed method had good sensitivity and selectivity for determining the ABA, EMA and IVM residues in vegetables within 12 min. When spiked at 10-1 000 μg?L-1, the fortified recoveries of ABA, EMA and IVM, respectively, were 90.5%-114.1%, 96.7%-118%, and 92.8%-108.1%, and their variation coefficients (C.V. %) were 0.85%-12.11%, 1.38%-9.36%, and 0.95%-11.52%, respectively. The limited detection levels of ABA, EMA and IVM residues in vegetables were 1.2, 2.1, and 1.7 μg?kg-1, respectively. 【Conclusion】 The HPLC-FD method is simple, fast and easy to perform, all validated parameters of this method are sufficient for analysis of pesticide residue.

    Analysis on Microstructure Difference of Ventral and Dorsal Side of Rice Grain

    LI Yi-nian,DING Wei-min
    Scientia Agricultura Sinica. 2010, 43(16):  3473-3480 .  doi:10.3864/j.issn.0578-1752.2010.16.026
    Abstract ( 1043 )   PDF (2125KB) ( 823 )   Save
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    【Objective】 Microstructure difference of rice between ventral side and dorsal side was researched, and the mechanical property difference with three-point breaking force between ventral side and dorsal side were also researched. Correlation between mechanical property and microstructure of rice was established, thus providing a theoretical basis for rice fissure appearance from dorsal side of rice grain. 【Method】 Ventral and dorsal side of two rice varieties at starch granule, cell and aleurone stratum levels were investigated under scanning electron microscope. Mechanical property of rice was tested by three-point breaking force. 【Result】 The microstructure of rice on ventral and dorsal side is different at starch granule, cell and aleurone stratum levels. Microstructure difference of rice on ventral and dorsal side affects structure strength and mechanical properties of rice between ventral and dorsal side. And the structure strength in ventral side is lower than that in dorsal side. These accord with mechanical property. Fracture form in microstructure is also different on ventral and dorsal side. 【Conclusion】 Rice fissure that started from the ventral side is related with structure strength of ventral side will be lower than that of dorsal side under desorption and absorption condition.