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Table of Content

    15 February 2011, Volume 44 Issue 4
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Plant Artificial Chromosome: The Vector for the Next Generation of Genetic Engineering
    LI Chen,YAN Xiao-hong,YANG Jie,YANG Qing,WEI Wen-hui
    Scientia Agricultura Sinica. 2011, 44(4):  657-663 . 
    Abstract ( 924 )   PDF (288KB) ( 1206 )   Save
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    The simultaneous expression of multiple genes and genetically modified (GM) food security research have been considered as the research focus for genetic engineering in plants presently. Unlike conventional gene transformation technologies, plant artificial chromosomes provide one solution to the stable expression of multiple transgenes. As plant artificial chromosome segregation is independent of host chromosomes, they provide a platform for accelerating plant breeding and for studying the specific chromatin domains inserted into them. The generation of artificial plant chromosomes and their applications were reviewed in this article.

    Improvement of Insect-Resistance of 9311 and Development of Insect-Resistant Rice Hybrids
    TIAN Yu,XU Jun-ying,LI Chun-hai,LIN Yong-jun,MOU Tong-min
    Scientia Agricultura Sinica. 2011, 44(4):  664-672 . 
    Abstract ( 822 )   PDF (364KB) ( 1392 )   Save
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    【Objective】To introgress insect-resistance gene cry1C* into 9311, an elite indica rice variety, for developing new rice lines and hybrids which are resistant against rice stem borers and leaffolders.【Method】T1C-19 harbouing cry1C*, a transgenic rice material, was used as donor of insect-resistance in the study. cry1C* was introgressed into the genetic background of 9311 by hybridizing, backcrossing and MAS. Improved new lines and hybrids were carried out for identifying insect-resistance in field, testing protein content of CRY1C*, evaluating yield, main agronomic traits, growth duration and rice quality in improved lines and hybrids.【Result】Eight near-isogenic lines (improved lines) with genetic background of 9311 and insect-resistance gene cry1C* were developed and 56 crosses were made. Identification in field showed that the levels of stem borer- and leaffolder-resistance were the same as that of the donor parent T1C-19. Improved new breeding lines and T1C-19 were not damaged by leaffolders, compared to the twenty percents in damage in 9311 in the same field. The expressing protein contents of CRY1C* in improved lines were the same as T1C-19. The expressing protein contents of CRY1C* of hybrids were lower than that of their male lines at tillering stage, but higher at elongation stage. The growth duration, main agronomic traits and rice quality of improved lines were the same as 9311, but the grain yield per plant was higher. Three insect-resistant combinations with long growth duration were screened for one cropping with higher yield over check hybrid Yangliangyou 6. Five insect-resistant combinations with higher yield and quality over check hybrid Q-you 6 were selected for one cropping with medium growth duration. One insect-resistant combination with the same yield and growth duration as check hybrid Jinyou 207 was selected for second cropping. 【Conclusion】 The insect-resistance gene cry1C* has been introgressed into the genetic background of 9311 by hybridization, backcrossing and MAS. The developed lines with insect-resistance have the same characters as 9311 in growth duration, yield, main agronomic traits, rice quality and combining ability. Several insect-resistant hybrids were selected for one or second cropping.

    Analysis on Genetic Diversity and Trait Association of Different Types of Azuki Bean (Vigna angularisi) by SSR Markers
    ZHAO Bo,YE Jian,JIN Wen-lin,ZENG Chao-wu,WU Bao-mei,PU Shao-jing,PAN Jin-bao,WAN Ping
    Scientia Agricultura Sinica. 2011, 44(4):  673-682 . 
    Abstract ( 820 )   PDF (716KB) ( 1144 )   Save
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    【Objective】 It is very important to make studies on the genetic diversity of wild, semi-wild and cultivated azuki beans for further detecting the origin, evolution and diffusion of azuki bean and effectively utilizing these germplasms and serving for azuki bean improvement. 【Method】 Eleven higher polymorphic SSR primers screened from 69 SSR primer pairs of azuki bean and black gram were used in amplification of 558 wild, semi-wild and cultivated azuki bean accessions from China, Japan, South Korea, Bhutan and Burma. The analysis of genetic diversity and trait association was done.【Result】 Eighty-six polymorphic bands were generated. An average of 7.28 polymorphic bands were detected from 1 pair SSR primers. The number of alleles changed from 6 to 10. There were specific bands were observed in wild, semi-wild and cultivated azuki beans. Specific bands of cultivated azuki bean were predominantly produced by Chinese cultivars and landraces. Specific bands of wild azuki bean were detected in accessions from China, Bhutan and South Japan. The order of genetic dispersion was wild>semi-wild>cultivated azuki beans. Semi-wild accessions has closer genetic relationship with wild azuki bean. The 558 accessions were divided into five main groups by cluster analysis according to genetic distance. The result of cluster appeared that the genetic types of azuki beans have obviously associated with different geographical regions. Japanese cultivated azuki beans have closer genetic relationship with wild and semi-wild accessions from Japan and South Korea. Wild accessions from Southwest China are closely related to wild accessions from Southeast Asia and landraces of Jiangsu Province. Trait association analysis showed that black gram BG111 marker located on linkage group 7 of azuki bean explained 49%, 44%, 31%, and 18% traits of number of nods on main stem, diameter of stem, top stem twining and branching numbers in wild and weedy types. BG48 of linkage group 1 and BG20 markers of linkage group 5 from black gram and azuki bean AZ24 marker of linkage group 9 accounted for 9%, 7%, 5%, and 6% traits of maturing days, plant height and seed number per pod, number of nods on main stem in cultivated azuki bean respectively.【Conclusion】 A higher genetic variation was detected in wild types than in cultivated accessions. Chinese cultivated azuki bean is originated from China and has the most diversely genetic variation. Number of nods, diameter of stem, top stem twining and branching numbers in wild and semi-wild types are related with black gram BG111 marker. Maturing days, plant height and the number of seeds per pod, number of nods on main stem are associated with black gram BG48 and BG20, azuki bean AZ24 markers in cultivated azuli bean, respectively.

    Construction and EST Analysis of the Normalized Full-Length Latex cDNA Library of Hevea brasiliensis
    ZENG Ri-zhong,DUAN Cui-fang,NIE Zhi-yi,DAI Long-jun,LI Yu
    Scientia Agricultura Sinica. 2011, 44(4):  683-690 . 
    Abstract ( 684 )   PDF (442KB) ( 895 )   Save
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    【Objective】Construction and normalization of the latex full-length cDNA library would facilitate the identification and analysis of the laticifer-specially expressed genes so as to gain insights into the molecular and regulatory mechanisms of the latex metabolism and the rubber biosynthesis that occur in the laticifers of rubber tree.【Method】Firstly, the primary full-length cDNA library was constructed with the latex of H. brasiliensis (clone Reyan 7-33-97) using recombination of the full-length latex cDNAs with the Gateway donor vector of pDONR222. Secondly, the plasmids of the primary cDNA library were normalized through self-subtraction with the Hevea genome DNA, and the normalized latex full-length cDNA library was constructed using the subtracted plasmids. 【Result】The titer of the unamplified primary cDNA library was about 6.0×106 cfu/mL. the average size of inserted cDNAs was 1.5 kb with a recombination of about 100%,and the ratio of full-length cDNAs was around 76%. The normalized latex full-length cDNA library had a total CFU of 1.87×105 with a recombination ratio of 95%. Quantity RT-PCR results demonstrated that normalization produced about 1 000 fold average reduction of two high abundant latex genes, i.e., small rubber particle protein (SRPP) and rubber elongation factor (REF). 【Conclusion】A high-quality latex normalized full-length cDNA library was successfully constructed, and 12 000 of random clones in this cDNA library were then sequenced. A total of 11 951 ESTs (expressed sequence tag) were attained, and could be assembled into 3 394 unigenes, including 2 061 contigs and 1 333 singlets. The latex cDNA library can be well used for Hevea’s functional genomics, novel gene screening, high-through EST sequencing and the preparation of the latex cDNA microarray of H. brasiliensis.

    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY·AGRICULTURE INFORMATION TECHNOLOGY
    Nitrogen Nutrition Diagnosis of Early Rice with NDVI and Its Application for Nitrogen Topdressing Recommendation at Yingtan, Jiangxi Province
    QIN Xia,WANG Shao-hua,XUE Li-hong
    Scientia Agricultura Sinica. 2011, 44(4):  691-698 . 
    Abstract ( 1044 )   PDF (259KB) ( 688 )   Save
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    【Objective】Establishment of the nitrogen uptake diagnosis model for early rice based on NDVI, and recommend- ation of the N topdressing rate. 【Method】 The quantitative relationship between canopy NDVI and plant nitrogen uptake (PNU) was studied using the data of field experiment with different N treatments, and the models were tested using independent experiment data. The optimum diagnosis model was established to recommend the N topdressing rate with canopy NDVI. Then the recommended N rate, grain yield and nitrogen use efficiency (NUE) were compared with those of farmer’s N management treatment.【Result】 Rice canopy NDVI was well positive correlated with PNU at key development stage, the correlation for single stage was better than whole stage, and exponential curve better fitted the relation than linear function, the same for the test results with independent experiment data. The prediction precision of PNU diagnosis model at tillering and jointing stages was 0.907 and 0.941, respectively. Compared with the farmer’s N management, equivalent grain yield was obtained with reduced N rate while NUE was improved. The error between the N rate of spectral diagnosis fertilization and the optimum N rate calculated with the N-yield curve was 2.12% at high transplanting density and -2.22% at low transplanting density, respectively, while the error for grain yield was -0.75% and -5.04%, respectively. 【Conclusion】 Spectral diagnosis fertilization mode can reduce nitrogen application rate and raise nitrogen use efficiency on premise of grain yield, and has a potential to be widely applied in rice N management.

    Comparison of Root Characteristics and Nitrogen Uptake and Use Efficiency in Different Corn Genotypes
    WANG Jing-feng,LIU Peng,ZHAO Bing-qiang,DONG Shu-ting,ZHANG Ji-wang,ZHAO Ming,YANG Ji-shun,LI Geng
    Scientia Agricultura Sinica. 2011, 44(4):  699-707 . 
    Abstract ( 835 )   PDF (303KB) ( 981 )   Save
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    【Objective】The aim of this study is to investigate the physiological mechanism of high nitrogen use efficiency of corn by comparing the differences in uptake and utilization of nitrogen, spatial and temporal characteristics of roots of summer corns with different nitrogen use efficiency (NUE). 【Method】Using different NUE corn varieties as materials, one is high NUE variety, LY13, and another is low NUE variety, LD981. The plant sample was separated to two parts, the aboveground (stem, leaf, grain) and underground (0-20, 20-40, 40-60, 60-100 cm root) part. At two nitrogen levels (0 and 4.29 g?plant-1), samples were collected at male tetrad, flowering, filling, milky and maturity stages to study their root spatial and temporal distribution, the difference in nitrogen uptake and utilization. 【Result】Compared with LD981, the root weight and root/shoot ratio of LY13 were maintained at significantly higher levels (P<0.05) post-anthesis. The domination in root spacing distribution, number of roots in deeper layer, root activity at later stage, and root functional period promote more absorption of nitrogen in LY13. The total nitrogen accumulation amount of LY13 overweighed LD981 significantly (P<0.05), which were more distinct at the high nitrogen level. This tendency could be found in nitrogen transportation efficiency, nitrogen contribution rate and nitrogen utilization efficiency (P<0.05). All these result that LY13 had superiority to LD981 (P<0.05) in grain yield, biological yield, 1000-kernel weight, seed set and harvest index at either nitrogen level. 【Conclusion】As for LY13, the higher nitrogen accumulation was mainly attributed to its more total root weight, more root in deeper layer, more rational root distribution, and longer duration of strong root activity; and the primary cause of its higher grain yield and nitrogen use efficiency were bigger seed set and stronger seed set adjustment ability promoting nitrogen translocation efficiency, leading to more nitrogen distributed to grain.

    Effects of Phenolic Compounds and Antioxidant Ability in Soybean Leaves Under O3 Stress
    ZHAO Tian-hong,JIN Dong-yan,WANG Yan,CAO Ying
    Scientia Agricultura Sinica. 2011, 44(4):  708-715 . 
    Abstract ( 869 )   PDF (444KB) ( 802 )   Save
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    【Objective】The objective of this experiment is to reveal the effects of phenolic substance contents and PAL, LOX activities in soybean leaves under elevated O3 concentrations, which will provide a basis at the molecular level for the adaptive mechanism of secondary metabolism in plant. 【Method】 Open-top chambers (OTCs) were utilized to investigate the effects of phenolic compounds and antioxidant ability in soybean leaves under different O3 concentration at (80±10 nmol?mol-1 and 110±10 nmol?mol-1) . 【Result】 The results indicated that, compared with control, the O3 concentration at 80±10 nmol?mol-1 induced an increase (P<0.05) of flavonoids content, polyphenols content and the activities of phenylalanineammonialyase (PAL), lipoxygenase (LOX), polyphenol oxidase (PPO). But the O3 concentration at 110±10 nmol?mol-1 induced an decrease (P<0.05) of flavonoids content and PAL activity. Then superoxide anion ( ) production rate, hydrogen peroxide (H2O2) content, relative electrical conductivity and malondialdehyde (MDA) content in soybean leaves increased under ozone stress. 【Conclusion】 The activities of PAL and LOX can be activated under certain extent elevated O3 concentration, which are beneficial to the accumulation of phenolic substances as to protect the normal growth and development of soybeans. But antioxidant system can not tolerate oxidative damage caused by elevated O3 concentration, thus increasing membrane lipid peroxidation, which represents injured effects on soybean.

    PLANT PROTECTION
    Establishment of REMI Mutagenesis of Setosphaeria turcica and Characterization of Mutants
    LI Po,GU Shou-qin,LIU Li-hua,NIU An-li,WU Min,DONG Jin-gao
    Scientia Agricultura Sinica. 2011, 44(4):  716-722 . 
    Abstract ( 727 )   PDF (341KB) ( 782 )   Save
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    【Objective】 The objective of this study is to establish high efficient REMI mutagenesis system, which is able to construct mutants that can be used to clone genes related to growth, development and pathogenicity of S. turcica.【Method】 REMI mutagenesis system was established through seeking the optimal different ages of this fungus, cell wall digest enzymes and its condition and collection method which affect protoplast yield, and seeking hygromycin B concentration for transformants screening, and seeking plasmid for transformation, which was used to achieve transformants. 【Result】 When lyallzyme, snailase, and drislase were mixed and at concentration of 1.25%, respectively, and when mycelium was cultured for 20 h, and 3 000 r/min for collection, the protoplast production was the maximum. Hygromycin B at 50 μg?mL-1 was determined as the optimal concentration for transformants screening, in which conidial germination and colony growth of wild type was completely inhibited. The plasmid pAN7-1 had higher transformation efficiency. The optimal system was used to induce mutagenesis and lots of transformants were obtained, from which some conidia production-changed and pathogencity-changed transformants were found. 【Conclusion】 The REMI mutagenesis of S. turcica was established, which had a great significance in construction of mutant library and in cloning the pathogencity-related genes.

    Screening of Laccase-Producing Isolates Among Plant Pathogenic Fungi
    ZHAN Xu,CAO Zhi-yan,XING Ji-hong,DONG Jin-gao
    Scientia Agricultura Sinica. 2011, 44(4):  723-729 . 
    Abstract ( 965 )   PDF (441KB) ( 1271 )   Save
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    【Objective】 Screening isolates from plant pathogenic fungi which be able to produce laccase and determining the laccase activity could give a deep insight into functional research for the pathogenicity of laccase. 【Method】 Using 2,2’-azino-bis[3-ethylbenzthiazoline-6-sulfonic acid] (ABTS) as substrate, isolates with laccase were screened from plant pathogenic fungi through the plate-color reaction and confirmed that these isolates had a role in lignin degradation using aniline blue (Azure-B). Laccase activity of six different isolates was compared using spectrophotometer at 420 nm. The effect of pathogenicity from laccase was analyzed using the histopathology. 【Result】 Ten isolates with a role in lignin degradation were screened from twenty plant pathogenic fungi. Setosphaeria turcica and Pestalotiopsis microspora contained the highest intracellular and extracellular laccase activity, which were 18.984 and 0.919 U?mL-1, respectively. The analysis of histopathology in S. turcica showed that maize leaves were oxidated by laccase which could accelerate the extention of pathogen against the host. 【Conclusion】 Most of the laccase in the plant pathogenic fungi were intracellular laccase which had a role in lignin degradation. In this study, laccase of S. turcica was found that could contribute to the expansion of pathogen in the host tissue.

    Cloning, Expression and Activity Analysis of Carboxylesterase Gene from Helicoverpa armigera (Hübner)
    LIU Xiao-min,LI Jie,GUO Wei,ZHANG Xia,LI Xin-na
    Scientia Agricultura Sinica. 2011, 44(4):  730-737 . 
    Abstract ( 714 )   PDF (944KB) ( 1051 )   Save
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    【Objective】 The objective of the study is to clone carboxylesterase gene and provide a molecular basis for insecticide resistance of organophosphates (OPs). 【Method】 A full-length cDNA clone was identified to code for a carboxylesterase by immuno-screening of the Helicoverpa armigera midgut cDNA expression library and expressed into Escherichia coli. The carboxylesterase activity of HC3 was characterized with the α-NA as a substrate and its enzyme activity of HC3 was analyzed. 【Result】 The hc3 is 2 896 bp in length, which encodes 795 amino acids (GenBank accession number is GU119888). The predicted isoelectric point of HC3 is 3.94. The HC3 possesses a catalytic triad, consisting of a Ser, a His and a Glu residue (Ser204, Glu331 and His444). The activity for HC3 is 0.32 mmol/100 μL enzyme buffer. 【Conclusion】 A cDNA encoding carboxylesterase was cloned and expressed from Helicoverpa armigera. A noval GDE hydrophilic region was found in HC3. All the above understanding will further explore the three-dimensional structure, hydrolysis of carboxylesterase and provide possibility for the design of new types of pesticides.

    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Effects of Tillage Systems on Fields in Chengdu Plain —II. The Evaluation of Soil Quality
    LIANG Shu-min,XIE Rui-zhi,LI Chao-su,YANG Jin-zhong,TANG Yong-lu,WU Chun,WANG Ling-bian,LI Shao-kun
    Scientia Agricultura Sinica. 2011, 44(4):  738-744 . 
    Abstract ( 661 )   PDF (246KB) ( 1068 )   Save
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    【Objective】 This paper evaluated the effects of the tillage systems on soil quality in Chengdu Plain. The soil quality evaluation system which is suitable to the local production conditions was explored and established. In order to provide reference for enhancing the local soil productivity levels and improving soil quality, the suitable tillage systems were selected.【Method】The soil properties of different tillage systems and their effects on the crop yield index were acquired by using long-term field tests. Using principal component analysis, the evaluation index and the soil quality were evaluated and screened by the factor analysis method.【Result】A significant difference was found in comprehensive evaluation of soil quality indicators in different soil layers. The main factors in 0-10 cm are capillary porosity, soil specific gravity, seepage rate of soil saturation, soil compaction and soil organic matter. The main factors in 10-20 cm are total porosity, non-capillary porosity, soil hydroscopic water and soil compaction. The main factors in 0-20 cm are non-capillary porosity, soil specific gravity, seepage rate of soil saturation and soil compaction. The same factors in different soil layers are seepage rate of soil saturation and soil compaction, which showed that they are the local limiting factors of soil quality currently. The results of soil quality assessment showed that wheat and rice no-tillage is the best tillage system in 0-10cm, wheat no-tillage and rice conventional tillage are the best in 10-20 cm and wheat no-tillage and rice conventional tillage are the best in 0-20 cm. 【Conclusion】 Under this experimental conditions and local conditions, the good efficiency tillage system for the soil quality in the Chengdu Plain is wheat no-tillage and rice conventional tillage system.

    Study on Yield Increasing and Nutrient Uptake Effect by Nitrogen Application and Nitrogen Use Efficiency for Winter Rapeseed
    ZOU Juan,LU Jian-wei,CHEN Fang,LI Yin-shui,LI Xiao-kun
    Scientia Agricultura Sinica. 2011, 44(4):  745-752 . 
    Abstract ( 684 )   PDF (358KB) ( 856 )   Save
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    【Objective】Field experiments were carried out to study the effect of nitrogen application on winter rapeseed (Brassica napus L.) in the Yangtze River Valley and further to explore approaches of increasing rapeseed yield and improving nitrogen use efficiency. 【Method】The indices, such as yield increment, increase rate, partial factor productivity (PFP), agronomic efficiency (AE), physiological efficiency (PE) and apparent recovery efficiency (RE) were calculated based on the data from 73 field experiments in 10 main rapeseed production provinces in the valley during 2004-2006, and the rapeseed yield increase from nitrogen application and nitrogen use efficiency were analyzed in the valley. 【Result】Application of N fertilizer increased winter rapeseed yield significantly, the average grain yield increment resulting from 180 kg N?hm-2 application was 1 109 kg?hm-2, and the increase rate was 71.7%. Although N application had little effect on harvest index, the shoot biomass of rapeseed was increased by 72.7%. Compared to the CK (no N) treatment, nitrogen, phosphorus and potassium uptake of +N treatment at harvest were increased by 90.0%, 55.4% and 59.8%, respectively. Nitrogen harvest index declined significantly and P and K harvest index were on the rise with N application. The results also showed that PFPN, AEN, PEN and REN of winter rapeseed were 14.7 kg?kg-1, 6.2 kg?kg-1, 18.5 kg?kg-1 and 34.6%, respectively. 【Conclusion】The combination of nitrogen with phosphorus, potassium and boron fertilizers could obviously improve the shoot biomass and nitrogen, phosphorus and potassium uptake, and thus increase the grain yield of winter rapeseed significantly. Nitrogen use efficiency for rapeseed production is still quite low, and it has a great potential for growth by improving the technology.

    HORTICULTURE
    Effects of Exogenous GABA on Reactive Oxygen Species Metabolism and Chlorophyll Fluorescence Parameters in Tomato Under NaCl Stress
    LUO Huang-ying,GAO Hong-bo,XIA Qing-ping,GONG Bin-bin,WU Xiao-lei
    Scientia Agricultura Sinica. 2011, 44(4):  753-761 . 
    Abstract ( 1083 )   PDF (322KB) ( 1008 )   Save
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    【Objective】The objective of this study was to investigate the effects of exogenous γ-aminobutyric acid GABA application on the alleviation of tomato injury of salt stress. 【Method】 Hydroponic experiments were performed to investigate the effects of exogenous GABA application (5 mmol?L-1) on growth, reactive oxygen species metabolism, chlorophyll contents, Pn and chlorophyll fluorescence parameters in seedlings of tomato cultivar ‘Jinpeng No.1’ under NaCl stress (50 mmol?L-1). 【Result】 It was found that exogenous GABA significantly increased activities of SOD, POD, CAT, GR and productive rate of , but slightly affected the growth, APX activity, contents of AsA, H2O2, MDA in tomato leaves under normal nutrient solution. On the other hand, exogenous GABA application to normal nutrient solution had no significant effects on chlorophyll contents, Pn, maximal photochemical efficiency of photosystem II (Fv/Fm) and non-photochemical quenching coefficient (NPQ), but Chl a/b, electron transport rate (ETR), actual photochemical efficiency (ΦPSⅡ) and photochemical quenching coefficient (qP) significant increased in tomato leaves. In comparison to NaCl stress, exogenous GABA application led to a marked increase in leaves growth, activities of SOD, POD, APX and GR, contents of AsA and GSH, as well as decrease in accumulation of , H2O2 and content of final product of membrane lipid peroxidation MDA, but little influence on CAT activity in leaves. Meanwhile, the addition of GABA to the salinized nutrient solution could maintain higher PSⅡ activity through significantly improving chlorophyll content, Pn, Fv/Fm, ETR, ΦPSⅡ and qP, as well as decreasing NPQ. 【Conclusion】Exogenous GABA application enhanced salt tolerance in tomato seedlings and alleviated the salt stress damage to PSII center via enhancing the antioxidant enzymes activities and the contents of antioxidants, reducing the accumulation of H2O2 and protecting the stabilization of membrane from peroxidation.

    cDNA Cloning and Expression Analysis of L-galactose-1-phosphate Phosphatase in Apple
    GUO Chun-miao,LI Ming-jun,MA Feng-wang,LIANG dong,ZHANG min
    Scientia Agricultura Sinica. 2011, 44(4):  762-770 . 
    Abstract ( 669 )   PDF (497KB) ( 728 )   Save
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    【Objective】This study is to understand the characteristics of apple L-galactose-1-phosphate phosphatase (GPP) and the relationship between its expression pattern and ascorbic acid (AsA) in apple. 【Method】 GPP cDNA was cloned from young fruit of Malus domestica Borkch cv. Gala, expressed in E. coli, and the specific antibody was prepared, then the relationship between AsA level and the expression level among different tissues of apple were analyzed.【Result】cDNA of GPP from young fruit of apple has an 813 bp open reading frame(ORF) and encodes a protein of 270 amino acid residues with a putative molecular mass of 29 kD. Its accession No. in Genbank is FJ752240. Both of its cDNA and amino acid sequence showed high similarity with GPP genes reported in other plants, but not with myo-inositol-1-phosphate phosphatase. After GPP was heterologously expressed in E. coli, the fusion protein (GPP-His, His was about 21 kD,and GPP was about 29 kD) was about 50 kD, and mainly showed as inclusion body. Then a polyclonal antibody was prepared in rabbit. By western blotting, the antibody could have a specific reaction with recombined protein of apple GPP. However,the size of GPP protein in apple was about 33 KD with protein extracted from apple tissues. Additionally, both of GPP mRNA and protein expression levels showed a good consistency with AsA content among different tissues of apple.【Conclusion】These results suggest that GPP protein in apple, as a monomer protein, has a character of post-translational modification, and its expression might play an important role in regulating AsA synthesis.

    Analysis of Ripening Characteristics, Aroma Components and Aroma-Related Enzymes in Oriental Sweet Melons (Cucumis melo var. makuwa Makino)
    QI Hong-yan,LI Yan,GUAN Xiao-chuan,LI Jin-ran,QIU Li-yan,HAO Jing
    Scientia Agricultura Sinica. 2011, 44(4):  771-780 . 
    Abstract ( 847 )   PDF (317KB) ( 1015 )   Save
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    【Objective】Comparison on ripening characteristics, main aroma compounds and aroma-related enzymes activities between two cultivars of oriental sweet melons were conducted.【Method】Gas chromatograph and oxygen electrode method were used to detect ethylene production and respiration rate, respectively. Varieties and content of aroma compounds were measured by gas chromatograph-mass spectrometer(GC-MS), whilst aroma-related enzymes activities were also determined.【Result】 Climacterics of ethylene and respiration of ‘Caihong7’ were 3 days earlier than ‘Japanese Tianbao’. The soluble solids contents of mature ‘Japanese Tianbao’ were significantly higher than that of ‘Caihong7’. The contents of alcohols and aldehydes were higher of ripening ‘Japanese Tianbao’ were higher than that of ‘Caihong7’ markedly, while the differences on content of total aroma volatiles and esters were not significant. The content of total and acetate esters reached maximum value on day 33 and 35 after anthesis in ‘Caihong7’ and ‘Japanese Tianbao’, respectively. Activities of LOX in flesh and peel of ‘Japanese Tianbao’ were significantly higher than in ‘Caihong7’, whereas AAT activities followed the opposite trend and no significant differences in ADH activities were observed in both cultivars. The content of aroma compounds and enzyme activities were higher in peel than in flesh of both cultivars. 【Conclusion】Higher contents of alcohols and aldehydes in mature ‘Japanese Tianbao’ considered to be important contributors to its cucumber-like flavor. The different components proportion of aroma compounds might be the reason for causing different types of fragrance between both cultivars. LOX, ADH and AAT had interaction effects on fragrance types.

    A SON-PCR Technique Suitable to Amplificating the Flanking Regions of Gene of Chinese Jujube
    XIE Nan-nan,QIAO Yong,ZHAO Jin,LIU Meng-jun
    Scientia Agricultura Sinica. 2011, 44(4):  781-788 . 
    Abstract ( 599 )   PDF (814KB) ( 588 )   Save
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    【Objective】In this experiment, Junzao was used as materials based on previous studies, to establish a SON-PCR technique for isolation unknown flanking sequence of Chinese jujube. 【Method】 The original two-stage second round of the SON-PCR reaction was improved to two-step: firstly, the PCR results of primary reaction were used as template, single nested primer was used to drive the 5′selected linear amplification; then, a small amount of the primer designed for primary reaction and primer designed for secondary reaction was added to specially amplificating secondary reaction. The specificity and efficiency were further enhanced, and the dilution of the original secondary reaction PCR template was adjusted to 10ng?μL-1 of the primary reaction result. 【Result】 Finally, the test data using the system to a 626bp fragment of 3 'flanking sequences were amplified to obtain a 934bp encoding leucine-rich repeat sequence related fragment. The universality of this technique was proved by using two other fragments.【Conclusion】The method can efficiently amplify unknown sequence flanking sequence.

    Reaserch of Protein Expression of Style In Self-Incompatibility Cultivar Prunus armeniaca L. cv. Xinshiji by 2D-DIGE Technique

    CAO Xiao-yan,FENG Jian-rong,WANG Da-jiang,BAI Ru,LIU Yue-xia
    Scientia Agricultura Sinica. 2011, 44(4):  789-797 . 
    Abstract ( 885 )   PDF (627KB) ( 785 )   Save
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    【Objective】 The objective of the experiment is to explore self-incompatibility (SI) molecular mechanism of apricot at proteome level.【Method】Different proteomics of style at the stage of the small bud, big bud, 24 h after self-pollination (ASP) and 24 h after cross-pollination (ACP) (‘Xinshiji’בBadanshui’) in the SI apricot ‘Xinshiji’ by two dimension difference gel electrophoresis (2D-DIGE) and mass spectrum (MS). 【Result】About 1500 protein points were detected, and 66 different proteins were expressed at four different development stage of style in ‘Xinshiji’. Self and non-self pollen had triggered different effects on style at proteomic level. There was one protein expressed special in cross-pollinated style, and four up-expressed and twenty-nine down-expressed in cross-pollinated style, respectively, compared with self-pollinated style. Thirteen protein spots were identified unambiguously.【Conclusion】 Different reactions on style at proteomic level had been obviously triggered in ‘Xinshiji’ by self and non-self pollen.

    Mechanism of Starch-Sucrose Metabolism Regulated by IBA as Well as GA3 During Scale Cutting Propagation in Lilium
    SUN Hong-mei,HE Ling,WANG Wei-wei,JIA Zi-kun,LI Tian-lai
    Scientia Agricultura Sinica. 2011, 44(4):  798-806 . 
    Abstract ( 941 )   PDF (360KB) ( 726 )   Save
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    【Objective】 The effects of IBA and GA3 on starch-sucrose metabolism during scale cutting propagation were studied with the aim to provide a theoretical and practical basis for regulating the development of bulblets and explore the physiological mechanism of scale propagation.【Method】The scales of Lilium Asiatic hybrids ‘Elite’ were immersed in IBA solution at concentration 200 mg?L-1 for 2 h and GA3 solution at concentration 200 mg?L-1 for 8 h, respectively. And the control was designed with water in same volume 2 h. The starch content, sucrose content and the activities of related enzymes in various positions of mother scales and bulblets were determined. 【Result】 Results showed that IBA and GA3 accelerated the starch degradation, sucrose synthesis and the transfer of carbohydrate from apical, central regions to basal region in mother scales. Furthermore, these substantial changes speeded up the bulblet morphopoiesis. Starch degradation was partly regulated by SP in parent scales, and the sucrose content was significantly positively correlated with the SS as well as SPS activity. The efficiency of IBA on carbohydrates metabolism was higher than that of GA3. Besides, IBA was conducive to synthesize sucrose earlier in the mother scales. By comparison, GA3 restrained the degradation of starch in the mother scales during the initial culture period and promoted more sucrose synthesis. 【Conclusion】 IBA and GA3 promoted the metabolism of starch as well as sucrose, and increased the activities of related enzymes in the parent scales, and then improved the propagation coefficient or produced the optimum bulblets.

    ANIMAL SCIENCE·RESOURCE INSECT
    Effects of Different F2 Schemes on QTL Mapping in Pigs
    GUO Yuan-mei,ZHANG Qin,REN Jun,ZHOU Li-hua
    Scientia Agricultura Sinica. 2011, 44(4):  807-813 . 
    Abstract ( 843 )   PDF (194KB) ( 549 )   Save
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    【Objective】To optimize the QTL mapping design in pigs, the effects of different F2 schemes on QTL mapping were investigated using a simulation method. 【Method】 Using a simulation method, 18 population structures were simulated to evaluate the effects of the numbers of individuals in F0, F1 and F2 generations, the ratios of males to females in F1 and F2 generations and the QTL frequencies in Fo generation on QTL mapping. 【Result】With the increasing number of F2 individuals, the power of QTL detection increased, and the 95% confidence interval narrowed, while the estimate of the addictive effect of the QTL was downward. The reason for the downward biased estimator of addictive effect was that the QTL alleles were not alternatively fixed in the two founder breeds. As the proportion of F0 females increasing, the 95% confidence interval reduced. The power and the estimates were improved with the difference of QTL frequencies increasing in the founder populations. 【Conclusion】The number of F2 animals, the proportion of F0 females and the QTL allele frequencies in Fo generation had significant effects on QTL detection, but the numbers of F1 and F0 individuals and the proportion of F1 females had no significant effect on the QTL mapping.

    Effects of α-Ketoglutarate on Muscle Energy Metabolism in Weanling Pigs Chronically Challenged with Lipopolysaccharide

    FU Da-bo,WANG You-wei,HOU Yong-qing,DING Bin-ying,WANG Lei,LIU Yu-lan,ZHU Hui-ling
    Scientia Agricultura Sinica. 2011, 44(4):  814-822 . 
    Abstract ( 830 )   PDF (418KB) ( 952 )   Save
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    【Objective】 The aim of the current study was to evaluate the effects of dietary supplementation with α-ketoglutarate (AKG) on muscle energy metabolism with reference to its molecular mechanism in weaned pigs after repetitive stimulation with lipopolysaccharide (LPS). 【Method】 Twenty-four piglets were assigned to a 2×2 factorial arrangements with LPS challenge (or without) and dietary supplementation of AKG at 0 and 1% as the main factors, total four treatments. The 24 weaned piglets (Duroc×Landrace×Yorkshire, 6.79±0.32 kg BW) were randomly allocated to one of the 4 treatment groups (6 replicates for each group). LPS challenged piglets were given a single intraperitoneal injection of LPS at 10, 13, 15 d and non-challenged piglets were injected intraperitoneally with physiological saline at the same doses. On 16 d, all pigs were killed and anesthesia and gastrocnemius tissues were excised. Muscle adenine nucleotide was assayed by HPLC, western blot assays were performed for muscle AMP-activated protein kinase (AMPK) and acetyl-CoA carboxylase (ACC) expression and their phosphorylation. 【Result】 There were significantly interaction effects of AKG supplementation and LPS challenge on AMP level and AMP/ATP ratio (P<0.05). Dietary supplementation of 1% AKG increased ADP (P<0.05), attenuated the increase of AMP level and AMP/ATP and the decrease of ADP and energy charge (EC) induced by LPS challenge (P<0.05). AKG supplementation and LPS challenge had no effects on the expression and phosphorylation of AMPK. Dietary supplementation of 1% AKG significantly inhibited ACC phosphorylation. LPS challenge significantly increased the expression and phosphorylation of ACC (P<0.05). 【Conclusion】These results demonstrated that, under the present experimental conditions, LPS challenge resulted in muscle energy depletion. Dietary supplementation of 1% AKG could maintain muscle energy metabolism. Dietary supplementation of 1% AKG might affect muscle energy metabolism via ACC signaling. AKG might affect muscle energy metabolism via ACC signaling, therefore improve body energy status after LPS challenge.

    VETERINARY SCIENCE
    Pharmacokinetic and Bioavailability Study of Tulathromycin in Swine
    ZHAO Yong-da,LIU Shuai-shuai,JIAO Xiao-jun,NI Heng-jia,YU Hui-min,HE Xiang-ren,WANG Si-fan,ZENG Zhen-ling,HUANG Xian-hui
    Scientia Agricultura Sinica. 2011, 44(4):  823-828 . 
    Abstract ( 1103 )   PDF (276KB) ( 825 )   Save
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    【Objective】The pharmacokinetics and bioavailability of the triamilide antimicrobial, tulathromycin, were investigated in swine. 【Method】Thirty pigs received 2.5 mg?kg-1 of tulathromycin injection by either intravenous (i.v.) or intramuscular (i.m.) route in three studies: study A (0 pigs, i.v.) ,study B (0 pigs, i.m, Huizhong, China.) and study C (0 pigs, i.m, Pfizer,USA.). Blood samples were collected and analysed by high-performance liquid chromatography (HPLC) with tandem mass pectrometry detection (LC-MS/MS) using ESI. Pharmacokinetic parameters were estimated using the WinNonlinTM software package and SPSS 16.0 analysis of the time and concentration data. 【Result】The tulathromycin concentration time data were fitted to noncompartment model. After i.v. injection plasma clearance (Cl) was 182.7 mL?h-1?kg-1, the volume of distribution at steady-state (Vss) was 17.2 L?kg-1 and the elimination t1/2 was 64.8 h. Statistical analysis on plasma showed that there were no signi?cant differences between pharmacokinetic parameters (P>0.05).Compared with tulathromycin (Pfizer,USA.), the relative bioavailability (Huizhong, China) following i.m. administration was 117%,the absolute bioavailability (Huizhong, China) following i.v. administration was 112%. It appeared that tulathromycin was rapidly absorbed , eliminated slowly and highly bioavailable . 【Conclusion】The results indicate that the two formulations are bioequivalent in both the rate and extent of absorption.

    Construction of Subtracted cDNA Library of the Early Growth Plate in Broiler Chickens with Thiram-induced Tibial Dyschondroplasia
    NING Guan-bao,TIAN Wen-xia,WANG Rui,QIN Ping,QIAO Jian-gang,LI Hong-quan,LI Jia-kui,BI Ding-ren,PAN Si-yi,GUO Ding-zong
    Scientia Agricultura Sinica. 2011, 44(4):  829-834 . 
    Abstract ( 722 )   PDF (570KB) ( 642 )   Save
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    【Objective】An experiment was conducted to construct subtracted cDNA library for selecting time series genes differentially expressed in the TD growth plate of broiler chickens at the early stage with cDNA microarray. 【Method】 AVIAN (AV) broiler chicks at 7 days of age were randomly divided into two groups. After fasting overnight, they were fed with regular diet (control) or the same diet containing 100 mg/kg thiram for 48h to induce TD (thiram diet-fed). Forward and reverse-subtracted cDNA libraries were generated by suppression subtractive hybridization technology (SSH). Identification of the inserted cDNA fragments in subtractive library was done using PCR. One hundred clones were randomly selected for further DNA sequencing, blast homology analysis and function prediction. 【Result】 A total of 2 227 positive clones were obtained and the size of inserts was between 200 bp and 1 000 bp. There were 97 homologous gene sequences shared more than 99% identity with genes known in chicken (Gallus gallus). Non-redundancy of sequenced genes was 68.7%. Meanwhile, 3 clones were found to be novel EST as no functional clues were associated with them by bioinformatic analysis. Most of these genes are involved in matrix formation, endochondral ossification and remodelling, developmental regulation, signal transduction, electron transport in mitochondrial respiratory chain and vascularization. 【Conclusion】Successfully produced cDNA library would make a good foundation for further printing cDNA microarray, screening differential expression genes of TD growth plates at different stages, and also may provide new insights into understanding the pathogenesis of TD.

    AGRICULTURAL ECONOMY & MANAGMENT
    Comparison Between Liquid Culture and Solid Culture Technologies for Edible Mushroom Spawn Production and Analysis on Their Economic Benefits
    GUO Jing-li,LIU Xu
    Scientia Agricultura Sinica. 2011, 44(4):  835-841 . 
    Abstract ( 1451 )   PDF (301KB) ( 837 )   Save
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    【Objective】Through comparing the profit ratio of cost of liquid culture and solid culture technologies for spawn production, the advantages of liquid spawn production were explored for providing strategies for facilitating the application of this technology to promote healthy and rapid development of edible mushroom industry.【Method】 In this paper, an edible fungus research laboratory and a large-scale production facility for edible mushroom were used as examples. Detailed records and statistics were kept for the production of Bai-Ling mushroom (pleurotus nebrodensis) spawn using liquid culture and solid culture technologies in the years of 2009 and 2010. Statistical analyses were carried out on the cost and the profit.【Result】Liquid culture technology for spawn production has significant profit advantages. For example, the time for spawn production is reduced by 24 days. The time for mycelia to fill a fruiting bag is reduced to 10 days. The contamination rate is reduced by 1.5%. The cost of one fruiting bag is reduced by 0.732 yuan. The profit ratio of cost is increased by 42.08%. However, many problems need to be solved before this technology can be broadly used. 【Conclusion】 Comparing with solid culture method, liquid culture technology for spawn production is obviously more advantageous, which is reflected in better spawn quality, reduction of contamination, increase of efficiency, reduction of cost and increase of production stability. For this technology to be broadly used, it is key to improve incubator shakers, fermentation tanks and inoculation equipments. Efforts to facilitate researches according to the demand of a dynamic market would promote the application of this technology in the fast-growing mushroom spawn production industry.

    RESEARCH NOTES
    Effects of Fertilization Measures of Nitrogen (N) on Grain Yield, Grain Quality and N-Use Efficiency of Midseason Rice in Hubei Province
    HAN Bao-ji,ZENG Xiang-ming,ZHUO Guang-yi,XU Fang-sen,YAO Zhong-qing,XIAO Xi-ming,SHI Lei
    Scientia Agricultura Sinica. 2011, 44(4):  842-850 . 
    Abstract ( 1006 )   PDF (247KB) ( 1056 )   Save
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    【Objective】 Four popular midseason varieties of rice (Luoyou8, Guohaozayou1, Fengyou22 and Jufengyou72) were used to study the effects of application technology of nitrogen (N) on grain yield, grain quality and N-use efficiency in Hubei province. 【Method】Field trials with five N treatments (CK, FFP, modified FFP, SHY and modified SHY) and four replications were conducted synchronously at both Chibi County and Jingmen County, Hubei province in 2008 and 2009, respectively. 【Result】The results showed that grain yield of the control with no N application at Chibi was significantly lower than that at Jingmen both in 2008 and in 2009, which implied that soil N fertility in Jingmen was higher than that in Chibi. Grain yield of modified farmers’ fertilizer practice(FFP), super high yield (SHY) and modified SHY were more than that of FFP at Chibi with yield increase rates 10.0%, 2.3% and 23.2%, respectively, in 2008; and 16.6%, 11.8% and 22.6%, respectively in 2009. However, at Jingmen, grain yield of modified FFP, SHY and modified SHY were lower than that of FFP in 2008 and slightly more than that of FFP in 2009 with yield increase rates 2.0%, 6.7% and 1.7%, respectively. Nitrogen agronomic efficiency (AE) and nitrogen partial factor productivity (PFP) of modified FFP and modified SHY were higher than that of FFP in Chibi. PFP of N of above two treatments was significantly higher, while AE slightly higher than that of FFP at Jingmen. N-use efficiencies (NUE) of SHY at two field trial spots in two years were far lower than that of other treatments. 【Conclusion】 Both modified FFP and modified SHY treatments achieved higher grain yield and higher NUE, which suggested that 20% reduction of nitrogen of FFP could increase grain yield as well as NUE. On the contrary, too much N application would not increase, even reduce grain yield. In addition, the optimized nitrogen management could also improve the grain quality of rice.

    Analysis of Glucoraphanin and Sulforaphane Contents in Different Organs of Broccoli
    YAO Xue-qin,XIE Zhu-jie,LI Guang-qing,QIU Hai-rong,LI Yuan
    Scientia Agricultura Sinica. 2011, 44(4):  851-858 . 
    Abstract ( 1063 )   PDF (216KB) ( 782 )   Save
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    【Objective】The anticarcinogenic activity of broccoli is most likely to be due to the activity of the isothiocyanates sulforaphane derived from glucoraphanin. In order to provide information in broccoli breeding for high nutrition quality, the goal of this study is to determine the contents of glucoraphanin and sulforaphane and analyze the correlation between them which is in the florets, stems and leaves of different genotypes of broccoli.【Result】Both glucoraphanin and sulforapane content difference was extremely significant in the florets, stems and leaves of different broccoli genotypes. The highest contents of them were both in the florets, while the lowest contents were in the leaves. The mean value of glucoraphanin content in the florets was 4.4- and 13.97-fold, respectively, higher than that in the stems and leaves, and the mean value of sulforaphane content in the florets was 4- and 8.7-fold, respectively, higher than that in the stems and leaves. However, compared to most materials, there were higher glucoraphanin contents in the stems and leaves of some broccoli materials about 50% of which in the florets. The correlation coefficient between the glucoraphanin and sulforaphane contents in the florets, stems and leaves of the ten different genotype broccoli materials were 0.9886**, 0.9994**, 0.9935**, respectively.【Conclusion】There exist some genotypic differences which can significant affect the glucoraphanin content in different genotype broccoli materials and organs. In the development and application of sulforaphane, the glucoraphanin contents can be the screening index. Broccoli materials which have higher glucoraphanin contents in the stems and leaves can be selected as the raw materials for extraction of the sulforaphane from the wastes.

    Polymorphism Analysis of H-FABP Gene and Correlation with IMF Content in Wei Pig
    ZHANG Chen-hua,WANG Yang,DING Yue-yun ,YIN Zong-jun
    Scientia Agricultura Sinica. 2011, 44(4):  859-866 . 
    Abstract ( 575 )   PDF (566KB) ( 823 )   Save
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    【Objective】 Polymorphism of heart fatty acid-binding protein (H-FABP) gene and its correlation with IMF content in Wei pig were studied. 【Method】 PCR-RFLP (HinfⅠ, HaeⅢ, MspⅠ3 kinds of restriction enzymes) and molecular marker techniques were used in the study and the genetic variation of H-FABP gene 5'-upstream region and second intron in 123 Wei pigs was analyzed. 【Result】 In 5'-upstream region, there was polymorphism in loci of the HinfⅠand the frequency of allele H was 0.6545, showing moderate polymorphism (PIC = 0.3500). In the second intron, polymorphism was detected in polymorphic loci HinfⅠ*, the frequency of allele B was 0.7195, showing moderate polymorphism (PIC = 0.3222). Loci of HaeⅢ also existed, allele D frequency was 0.6301, showing moderate polymorphism (PIC = 0.3575 ). While no polymorphism was detected in the loci of MspⅠ, which the genotype was AA genotype. The gene frequencies and genotype frequencies of pigs tested have reached the Hardy-Weinberg equilibrium (P>0.05). The detection of the genotype of the IMF content had the trend of HH>Hh>hh, dd>Dd>DD, BB>Bb>bb, genetic effect values were as following: 4.6918, 4.2665, 3.7712, 4.6124, 4.3167, 3.8173, 4.3042, 4.2358, and 4.1970. Cloning and sequencing analysis on polymorphic fragments of three PCR-RFLP in H-FABP gene showed that HinfⅠ-RFLP is due to T→C mutation in 1 324 bp, HaeⅢ-RFLP is due to C→G mutation in 1 811 bp, HinfⅠ*- RFLP is due to T→C mutation in 1 970 bp. Sequencing results had 97.9% and 98.3% homology with the sequence X98558 and Y16180 published by GenBank. 【Conclusion】Polymorphisms in 5'-upstream region and the second intron of H-FABP gene had certain effect on IMF content in Wei pig. However, whether these polymorphisms are important genetic markers, which are related with H-FABP gene and IMF content in Wei pig, are needs to be further studied.