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Table of Content

    15 February 2010, Volume 12 Issue 1
    Research Progress on Cytoplasmic Male Sterility and Fertility Restoration in Gossypium hirsutum L.
    HOU Si-yu, ZHANG Rui, ZHANG Xiao, GUO San-dui
    2010, 12(1):  1-7.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Cytoplasmtic male sterile (CMS) has plays an important role in heterosis usage in cotton. CMS and utilization of fertility restorer system have made a breakthrough in conquering breeding bottleneck by artificial emasculation and unfolded a bright outlook for promoting commercial production of hybrid seed. This paper expounds the research and utilization of cytoplasmic male sterility and fertility genetics; the development of molecular markers linked with CMS-related mitochondrial genes and fertility restoring gene; fine mapping of fertility restoring genes and cloning of fertility-related genes. The paper also points out existing problems and prospected researched direction in the future.

    Application and Prospect of RNA Interference in Cotton
    YIN Guo, ZHANG Chao-jun, LI Fu-guang
    2010, 12(1):  8-15.  DOI: 10.3969/j.issn.1008-0864.2010.
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    RNA interference (RNAi) technology, with the characteristics of simplicity, high efficiency and high-throughout, has been paid more and more attention, and become a research hotspot in the field of biotechnology, such as identification of gene function. This paper briefly introduces the characteristics and latest achievements of this technique in plants, and expounds its application in cotton research field.

    Roles of Plant miRNAs in Adaptive Responses to Soil Stresses
    YANG Cun-yi, HUANG Lan-lan, ZHAO Mo-ran, NIAN Hai
    2010, 12(1):  16-22.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Soil with deficient basic nutrient or toxic elements is widespread globally and thus seriously affects crop production. To reduce the damage caused by soil stress, plants have evolved their adaptive mechanisms. Recent studies have found that miRNAs possess important roles in plant response to soil stress. This paper summarizes the roles of miRNAs in responding to soil stress of nutrients deficiency and toxicity elements, lays emphasis on integration of miRNAs in soil stress regulatory networks, and puts forward thoughts for improving plant adaptability to problem soils.

    Research Progress on Genetics and Molecular Mechanism of Starch Gelatinization Temperature of Rice Grain
    XIAO Peng, SHAO Ya-fang, BAO Jin-song
    2010, 12(1):  23-30.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Gelatinization temperature (GT) is an important quality predictor that determines the cooking quality of rice. Low GT rice needs less energy input during cooking than high GT rice. The structure of amylopectin, especially its side-chain length distribution, has close relation to GT. The more amount of short chains, the lower starch GT. The quantitative trait locus (QTL) mapping has identified the alkali degenerate (alk) gene located on chromosome 6 as a major gene underlying the GT, which encodes the starch synthase IIa (SSⅡa). Functional single nucleotide polymorphisms (SNPs) found in SSⅡa are closely associated with GT and amylopectin structure. These SNPs are being used for marker assisted selection to improve rice quality. A molecular mechanism to control the variation in gelatinization temperature of rice starch by SSⅡa has also been put forward.

    Application Prospects of Proteomics in Detection and Study of Genetically Modified Organisms
    PAN Ying-hong
    2010, 12(1):  31-34.  DOI: 10.3969/j.issn.1008-0864.2010.
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    The existing conventional methods for detection and study can not meet the requirements of rapid, accurate, sensitive and complete analysis of genetically modified organisms. It is quite necessary to apply proteomics to improve these methods. This paper briefly introduces the existing conventional technology for analyzing genetically modified organisms and  discusses about the application prospects of some important proteomics technologies in detecting and studying genetically modified organisms.

    Implementing Good Agricultural Practice Management to Improve Milk Quality and Safety
    ZHANG Jun-min1, ZHANG Shu-yi2, DENG Li-qing1, ZHAO Qing-yu1,WANG Jia-qi1
    2010, 12(1):  35-39.  DOI: j.issn.1008-0864.2010.05
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    Milk quality and safety have aroused extensive attention by our government, enterprises and consumers. Implementing good agricultural practice (GAP) management is one of the important measures ensuring the raw milk quality and safety. This article expounds the present status of dairy industry, dairy GAP application, and significance of implementing GAP certification in China. The paper also provides suggestions according to problems existing in GAP implementation.

    Research Progress and Thoughts about Proper Exploitation of Field Water Potential Productivity in Farming-pastoral Ecotone of Northern China
    PAN Zhi-hua1,2, ZHANG Lu-yang1,2| CHEN Dong-dong1,2, ZHOU Meng-meng1,2
    2010, 12(1):  40-45.  DOI: 10.3969/j.issn.1008-0864.2010.
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    The important reason for ecological degradation in northern farming-pastoral ecotone is unreasonable and too intensive utilization of field water. Proper exploitation and utilization of field water is the key to develop agricultural production and restore ecosystem. This paper analyzed the research progresses in field water situation, the production potential, and new tasks facing the exploitation of field water potential. It also pointed out that the development and utilization must follow the principle for proper and rational usage. The basic connotation of proper exploitation of field water potential was identified preliminarily. The research framework and key scientific topics were put forward for the farming-pastoral ecotone of northern China. At present, we should guide the local agricultural production by the above mentioned principles, to restore ecosystem and to promote the sustainable development of northern farming-pastoral ecotone.

    Research Progress and Prospect on Alternative Irrigation Technology of Divided Root
    CHAI Qiang
    2010, 12(1):  46-51.  DOI: 10.3969/j.issn.1008-0864.2010.
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    This paper expounds the alternative irrigation technology about its emerging background, operation modes, application efficiency and water-saving mechanism, and prospects the key research fields of this technology in the future. Alternative irrigation technology can significantly increase the water use efficiency (WUE) of crops, improve the quality of products, create a special root system favorable for improving crop WUE, and remarkably reduce evaporation speed under minor decrease of photosynthesis speed. This technology can also optimize the biochemical adjusting organism to drought resistance and improve WUE, and is favorable for improving moisture compensation and nutrients utilization efficiency, and decrease useless evaporation and leakage. The main research orientation in the future will include the adjusting role and mechanism of alternative irrigation to crop procreating process, the  integrated application and mechanism of this technology in intercropping and high density planting.

    Suggestion and Discussion on Construction of South China Crop Breeding Base
    DONG Zhao-hui, ZHANG Ying-lu, LIU Ji-fang, WANG Fei-jie
    2010, 12(1):  52-55.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Being an unique crop breeding method firstly initiated by China, “Nan Fan” has made special contributions to crop breeding for shortening breeding process, and accelerating generation reproduction. So far, about 80% of over 5 000 varieties all over the country have been bred by  “Nan Fan” crop breeding method. On the basis of summing up present status of “Nan Fan”, in order to take advantages of “Nan Fan” to speed up the cultivation of new crop varieties, and to promote stable development of grain and agriculture production, this paper analyzed the existing problems limiting its development  in planning, management, infrastructure construction and land use conflicts for “Nan Fan”. It also put forward recommendations accordingly.

    Thoughts about De-concentrated Spatial Layout of National Agricultural Science and Technology Park
    LI Xiu-bin
    2010, 12(1):  56-59.  DOI: 10.3969/j.issn.1008-0864.2010.
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    The functioning modules spatial layout of certain national agricultural science and technology parks are too dispersive. This phenomenon has resulted in far distances between bases of technology innovation, experiment, technic service, production, processing and logistics. Thus, the integrated innovation can not be put in to full play. Arising of these problems mainly dues to the lacking of knowledge as the Park is a “space measure” for integrated innovation and regional agricultural development. The agricultural science and technology park is a model for developing modern agriculture, rather than conventional agriculture. While modern agriculture is characterized by its spatial concentration, which is one of the basic requirements for the Park as an agricultural integrated innovation model at present stage. During the construction of the Park, we should clarify its position as a form of modern agriculture. The spatial layout of those core functioning modules within the Park should be properly concentrated.

    Studies on Genetic Relationship Analysis of 16 Phalaenopsis hybrid Cultivars by ISSR Molecular Marker Technology
    LI Min, WANG Yao-feng, MING Feng
    2010, 12(1):  60-65.  DOI: 10.3969/j.issn.1008-0864.2010.
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    By optimizing DNA extraction method, annealing temperature, amplification of cycle number and Mg2+ concentration, we established an optimal ISSR-PCR reaction system for Phalaenopsis in this study. 8 primers were used to generate 193 brands of DNA fragments, the average percentage of polymorphic bands was 92.2%, and the range of amplified fragments was 300~2 500 bp.  This result indicated that the tested species possessed abundant genetic polymorphism. The 16 species of Phalaenopsis can be divided into 2 groups at L=0.222 5, one stand for Japanese/Taiwan species, and the other is for species newly cultured by the Company. The potential hybrid combinations of different species were suggested according to the affinity distance determined by molecular marker.  The establishment of ISSR molecular marker system in Phalaenopsis has provided theoretical basis for analyzing genetic relationship of Orchidaceae ornamental plants.

    Studies on Edible Membrane Property of Soy Protein Isolate and Cellulase Modified Carrot Fiber
    LIU Huan, WANG Xin-wei, SUN Xiu-xiu, LIAN Zi-xuan, LV Hong-ju, LI Ming, ZHONG Ti
    2010, 12(1):  66-71.  DOI: 10.3969/j.issn.1008-0864.2010.
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    In this paper, edible membranes made from soy protein isolate (SPI) and cellulase modified carrot fiber (CF) were taken as material to study the effect of three factors: CF content, cellulase concentration, enzyme reaction time, on edible membrane property; to analyze the composite reaction of SPI and CF and the mechanism of celluase modified CF. The result showed that the 3 factors had significant impact on edible membrane properties (P<0.05). With the increase of CF content (0~25%), the tensile strength (TS) increased and elongation (E) decreased, while permeability index (WVP) firstly decreased and then increased. When CF content reached 16.7%, WVP reduced to minimum. With the augmentation of cellulase concentration or prolongation of cellulase reaction time, TS and E firstly increased, then decreased, and WVP decreased. When cellulase concentration reached 0.2 IU/g, or cellulase reaction time was 1 h, TS and E were maximum and the change of WVP became stable.

    Studies on Polymorphisms Within Intron 1 and Exon 2 of MBL1 Gene in Three Cattle Breeds
    LIU Mei1,2, JU Zhi-hua1, LI Qiu-ling1, WANG Hong-mei1, HUANG Jin-ming1,LI Jian-b
    2010, 12(1):  72-78.  DOI: 10.3969/j.issn.1008-0864.2010.
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    The single nucleotide polymorphisms (SNPs) within intron 1 and exon 2 of MBL1 gene in Chinese Holstein, Luxi cattle and Bohai cattle were investigated by using nested-PCR, DNA sequencing technology and CRS-PCR method. Sequencing results showed that three new SNPs were identified. They were 855(G/A)、2651(G/A) and 2686(T/C). 855(G/A)SNP was located in intron 1; 2651(G/A)caused amino acids alteration in Val24Ile; 2686(T/C)was synonymous mutation. Predominant allele were identical at three SNP sites in three cattle populations, which were G, G, C and the allelic frequencies were 0.87/0.58/0.57, 1/0.75/0.74, 1/0.76/0.63, respectively. By χ2 tests, we found that 855(G/A) of Chinese Holstein, 855(G/A) and 2651(G/A) of Luxi cattle and all sites of Bohai cattle were in accordance with Hardy-Weinberg equilibrium (P>0.05). The value of polymorphism information indicated that 855(G/A) was slight polymorphism; 2651(G/A)and 2686(T/C)were moderate polymorphism in the three cattle breeds(0.25<PIC<0.5).

    Stability Comparison of cytb and β-actin Genes Expression in Sea Cucumber (Apostichopus japonicus)
    YANG Ai-fu, ZHOU Zun-chun, DONG Ying, JIANG Bei, WANG Xiao-yu, CHEN Zhong, GUAN
    2010, 12(1):  79-84.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Analysis of gene expression commonly uses internal control gene for normalization. The mRNA levels for cytb and β-actin genes were detected by using RT-PCR at 11 larval development stages of sea cucumber(Apostichopus japonicus)including unfertilized egg, fertilized egg, cellulous stages, blastula, gastrula, early auricularia, auricularia, late auricularia, doliolaria, pentactula, juvenile and in different tissues of young sea cucumbers including body walls, coelomocytes, intestines and respiratory trees. The results showed that cytb gene expression was stable at all development stages and in the four tissues. The mRNA levels for β-actin gene showed significant differences at different development stages, but were stable in the tissues of body walls, intestines and respiratory trees. Moreover, cytb mRNA showed no significant changes in expression after a LPS challenge at the stages of gastrula, early auricularia, auricularia, late auricularia, doliolaria and juvenile or in the four tissues of young sea cucumbers. The expression of β-actin gene was also stable in the tissues of coelomocytes, intestines and respiratory trees after a LPS challenge. This study provides information for the selection of cytb gene and β-actin gene as suitable internal control gene in target genes expressions research of sea cucumber.

    Index Screening and Comprehensive Evaluation for Drought Resistance of Peanut Varieties at Germination Stage
    ZHANG Zhi-meng1, WAN Shu-bo2, DAI Liang-xiang1, SONG Wen-wu1, CHEN Jing1, MIAO H
    2010, 12(1):  85-91.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Under PEG solution simulated soil drought condition, the experiments were conducted to study on the response to water stress and estimate the drought resistance of twenty-seven varieties at germination stage. The results showed that the characters of peanut response to water stress were different among every variety at germination stage. The response of root length, rooting ratio, root dry weight and germinating rate of these varieties to water stress treatments were obviously different. These indexes were significantly or very significantly related to colligate drought resistance. We found 8130, Jihua No.2, Jihua No.4, Huayu No.27, Huayu No.25 and Luhua No.14 had better drought resistance. These results came from field observation and inside of laboratory the bourgeon method with high PEG concentration solution is a quick, simple and accurate method for drought-resistance identification.

    AFLP Genetic Map Construction and QTLs Identification from F2 Reciprocal Populations in Common Carp, Cyprinus carpio L.
    WANG Jun, WANG Cheng-hui, LIU Hao, MA Yu-qing, YANG Xin-xin
    2010, 12(1):  92-99. 
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    Taking F2 reciprocal populations derived from a diallel cross of Purse red common carp and Xingguo red common carp as material, we constructed genetic maps in the common carp (Cyprinus carpio L.) using AFLP marker and identified the growth-related quantitative trait loci (QTL). In positive mating population, 542 polymorphic markers were produced from 14 pairs of AFLP primers, in which 325 markers were accord with the ratio of 3∶1. A genetic map containing 50 linkage groups were constructed with genome length of 3 676.1 cM, and 17 QTLs related to total length, standard length, pre-dorsal height, caudal peduncle length and caudal peduncle height were achieved, which could explain 1.66% to 70.49% of the total variation. In negative mating population, 605 polymorphic markers were produced with 14 pairs of AFLP primers, in which 333 markers were accord with the ratio of 3∶1. Another genetic map containing 50 linkage groups were also constructed with genome length of 3 943.1 cM, and 15 QTLs related to body weight, total length, standard length, pre-dorsal height, caudal peduncle length were also gained, which could explain 1.58% to 63.89% of the total variation. Furthermore, this paper carried out initial analysis and discussion on genetic maps constructed from reciprocal population, the differences between QTL identification and their results.

    Applications of Pi-z、Pi-ta2 Genes in Japonica Rice Breeding with Blast Resistance |in Yunnan
    ZHANG Xue-tang, LIAO Xin-hua, ZHU Zhen-hua, SU Zhen-xi, SHI Rong, ZHAO Guo-zhen,
    2010, 12(1):  100-105.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Japan strains was used as standard varieties and resistance identification method was synchronized with breeding, the performance of Pi-z, Pi-ta2 genes was analyzed  in blast resistant rice breeding in Yunnan from 2002 to 2007. The result showed that till 2007 Pi-ta2, Pi-z and Pi-zt genes were still of utilization value in rice producing reigns of Yunan, and resistance of Pi-ta2 had distinct inconformity between leaf resistance and spike resistance to blast, while the other two had the same performance. Pi-z had more obvious effect in improving resistance of breeding materials in different resistant breeding combinations.

    Effects of Overexpression and RNAi-mediated Silence of OsCtBP-A Gene on Root Growth and Stress Resistance in Rice
    WU Mao-sen, LI Lei, WU Jing, CHEN Hua-min, HE Chen-yang
    2010, 12(1):  106-110.  DOI: 10.3969/j.issn.1008-0864.2010.
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    In order to expound the functions of cloned C-terminal binding protein gene OsCtBP-A, we established its overexpression and RNAi constructs and transformed them into calli of rice (cv. Nipponbare) via Agrobacterium-mediated transformation. Forty-five transgenic plants with overexpression and 75 with RNAi were generated and identified by PCR and GUS staining analysis. In T1 progeny of transgenic rice lines with overexpression, OsCtBP-A gene expression was remarkably increased, and root development was enhanced during seedling stage, while its resistance to rice leaf blight was no much changed. In contrast, OsCtBP-A gene expression was significantly reduced, and the root development at seedling stage was slowed down and its resistance to drought was weakened in RNAi silenced lines. The results suggested that OsCtBP-A gene might be involved in regulation of root development at seedling stage and resistances to drought and diseases.

    Cloning and Sequence Analysis of Heat Shock Protein 70 Gene from Ageratina adenophora(Eupatorium adenophorum)
    XIAO Yan-ping1,2,3, GONG Wei-na2, WAN Fang-hao2, LI Zheng-yue1, XIE Bing-yan3
    2010, 12(1):  111-117.  DOI: 10.3969/j.issn.10080864.2010.
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    Heat shock protein 70 (HSP70) plays an essential role in plant stress resistance. According to the conserved regions of hsp70 from other plant species, a pair of specific primers were designed. A fulllength cDNA of hsp70 gene from Ageratina adenophora was cloned by the technique of RTPCR. Its ORF (open reading frame) (GenBank locus: FJ598132) named hsp70.58 was 1 938 bp. It encoded a polypeptide of 645 amino acids with an estimated molecular mass of 70.58 kDa and contained the three highly conserved HSP70 family signatures and Cterminal pentapeptide EEVD. Relative realtime RTPCR was used to analyze the expression of hsp70.58 gene after heat shock and cold shock, respectively. The results showed that mRNA expression of hsp70.58 was significantly upregulated after temperature stress. It indicated that this gene belonged to hsp70 inducible type.

    Expression of ChIFN-&alpha|in Transgenic Tobacco Causes Resistance to Tobacco Mosaic Virus
    SONG Li1, ZHAO De-gang 1,2, TIAN Xiao-e1, WU Yong-jun2
    2010, 12(1):  118-122.  DOI: 10.3969/j.issn.1008-0864.2010.
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    Alpha interferon is a pleiotropic animal cytokine that possess powerful and wide-range of antiviral properties. The expression vector carrying chicken alpha interferon gene (ChIFN-α) driven by 35S promoter has been introduced into Nicotiana tabacum L. Xanthin by Agrobacterium transformation in our work. The transgenic plants were detected by Southern blot and Western blot. Recombinant ChIFN-α protein in tobacco leaf was analyzed by enzyme-linked immunosorbent assay (ELISA). The results of tobacco mosaic virus (TMV) challenge test showed that transgenic tobacco possessed certain resistance to TMV infection. And this might be related to the up-regulated expression of PR-la、N and MAPK genes in transgenic plants after TMV infection.

    Effects of Hyper-gravity on Physiological and Biochemical Indices of Tomato Seedlings
    DUAN Zhi-ying, YANG Zhi-fen
    2010, 12(1):  123-127.  DOI: j.issn.1008-0864.2010.01.20
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    Taking tomato (Lycopersicon esculentum Mill) seeds as plant material, this experiment determined the seed germination rate and index, chlorophyll content, root activity, catalase activity (CAT), peroxidase activity (POD), malondialdehyde (MDA) and proline content, and compared the results with those of the control, after they were treated with 1 000×g, 1 500×g, 2 000×g for 1, 2, 3 hours. The results showed that both germination rate and germination index decreased after hyper-gravity treatment, while the root activity, CAT and POD all rose, but the MDA content decreased. And the chlorophyll content increased after tomato leaf blades were treated with 1 000×g, 1 500×g for 1 hour and 1500×g for 2 hours. The free proline content of tomato seedlings increased, except they were treatment with 2 000×g for 3 hours. The above results indicated that different hyper-gravity treatments could enhance stress resistance of tomato seedling.

    Establishment of an Efficient Agrobacterium-mediated Transformation System for Genetic Improvement of Alfalfa (Medicago sativa L.)
    XIE Xin-xing1, LU Yang2, LIANG Jing2, MA Cui1, HE Yi-kun2
    2010, 12(1):  128-134.  DOI: 10.3969/j.issn.1008-0864.2010.
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    By comparative analysis on the different explant tissues, concentrations of bialaphos for selection, pre-culture and inoculation time, etc, an efficient Agrobacterium-mediated transformation system for an elite alfalfa (Medicago sativa L.) cultivar Debao was established. The results showed that cotyledon was the best explant for callus induction and transformation. The optimal bialaphos concentration for selection of transgenic calli and plants was 2.5 mg/L in the selection medium. The pre-culture of cotyledon before inoculation with Agrobacteria cells was a key factor affecting transformation efficiency, ranging from less than 5% without pre-culture to about 30% with 3 days of pre-culture. Three days of inoculation was optimal for transformation. More than one hundred independent transgenic plants were obtained, 30 were randomly selected for PCR amplification of bar gene and all the 30 plants were confirmed to be transgenic. The PCR positive plants survived and grew well after spray of 5‰ Basta solution. The Agrobacterium-mediated transformation system described in this study was proved to be reliable and efficient.

    Method for Rapid Measuring Cis-epoxysuccinate Hydrolase Activity
    PENG Liang-liang1,2, WANG Zi-qiang2, WANG Yun-shan2, ZHANG Li-ping1, SU Zhi-guo2
    2010, 12(1):  135-138.  DOI: 10.3969/j.issn.1008-0864.2010.
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    On the basis of cis-epoxysuccinate hydrolase (ESH) producing cell ultrasonication, orthogonal experiments were conducted to analyze the effect of substrate concentration, the enzyme concentration and the time of biotransformation on the activity of ESH in the process of L(+)-tartaric acid production with cis-epoxysuecinate as the substrate for fermentation. An extraordinarily faster method for measuring ESH activity was constructed, which was to put 1 g wet cells into 30 mL 0.2 mol/L ES aqueous solution under the condition of 37℃ for 10 min to biotransform and then to measure the concentration of L(+)-tartaric acid with HPLC. This method possesses the advantages of fast speed, high accuracy and small quantities of cells which were suitable for strain selection, inspection of fermentation parameter, optimization of fermentation conditions and industrial production.