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    15 April 2013, Volume 15 Issue 2
    The Progress and Perspective of Rice Functional Genomics Research
    XIAO Jinghua, WU Changyin, ZHANG Qifa
    2013, 15(2):  1-7.  DOI: doi:10.3969/j.issn.10080864.2013.02.01
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    Rice is a staple food crop and model system for genomic research among cereal plants. There has been rapid advances in rice funciotnal genomic research in the last decade including development of technological and resource platforms and the isolation of functional genes. Highthroughput genomic technologies have been used in rice breeding. This review gave a glimpse on the progress made in rice functional genomics research, and the perspective of rice development direction in the future around a goal referred to as “Rice 2020”: a call for an international coordinated effort in rice functional genomics.

    A Draft Sequence of Wheat D Genome and Its Perspectives in Application
    JIA Ji-zeng, KONG Xiu-ying, ZHAO Guang-yao, GAO Li-feng
    2013, 15(2):  8-13.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.02
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    Wheat is now the first food crop all over the world, and its wide spread adaptation and improved quality are mainly due to the integration of D genome into tetraploid wheat. Aegilops tauschii is the donor of D genome in common wheat. Here, we introduced briefly the structure of D genome, then primarily focused on the application of wheat genome sequencing on wheat studies, such as gene discovery, germplasm diversity and haplotyping, variety improvement, comparative genomics, genome evolution and polyploidization. We also gave some suggestions for future researches on wheat genome in China.

    Progress and Prospect of Genomics Technologies in Horticultural Crops
    WANG Xiao-wu
    2013, 15(2):  14-19.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.03
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    China is the largest growing and consuming country of horticultural crops in the world. The second generation of sequencing technologies achieved significant breakthrough and greatly influenced improvement of horticultural crops. A number of important horticultural crops, including Chinese cabbage, cucumber, tomato, melon and water melon et al. have recently being sequenced. After analyzing the progress of the genomics technology and policies adopted by the major contrived in the world, the paper listed some major tasks for promoting genomics of horticultural crops in China and proposed some policy recommendations.

    Study Progress of Gossypium Polyploidization
    WANG Kun-bo1, LIU Xu2
    2013, 15(2):  20-27.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.04
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    Polypoidization plays a key role in plant speciations and their evolution, the genus Gossypium, as a good example. Gossypum is a big genus in crops. There might exist polyploidy processes in most cotton species and many diploid cotton specie are a kind of polyploid which kept diploid chromosomes but existed polploidization. Gossypium bickii is a typical one and G. gossypioides is very special in the genus. The polyploidy indued species include G. aridum  and three ones of B genome. In Gossypium allotetraploid speies is a typically polyploid origination. G. raimondii was generally considered as the closest donor of allotraploid cotton D-subgenome s but was not taken so at least to the G. hirsutum D-subgenome verified with gDNA-FISH. The allotraploid cotton sepeciation took place perhaps in Mid-Pleistocene or a little later. Recent FISH studies on cotton contributed much to speciations, interspecific relationships, and evolutions in Gossypium. There will be great advantages of FISH and genomics including cp-genome in studies on cotton polyploidization.

    The Applications of Metabolomics in Plant Biology——Current Status and Prospective
    ZHANG Feng-xia, WANG Guo-dong*
    2013, 15(2):  28-32.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.05
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    Plants collectively produce more than 200 000 organic chemicals (molecular weight <1 000), which exhibit diverse physiological functions during the life cycle of plants. Thus, plant metabolomics now plays a fundamental role in plant biology and applied biotechnology. The association analysis of metabolomics and transcriptomics data (or other omics data) is a powerful tool for elucidating the function of genes involved in plant metabolic network. In recent years, plant metabolomics exhibits new potentials to new research fields such as fluxome and cofactome analysis. We briefly discussed the current status and the applications of metabolomics as a plant functional genomics and system biology tool in this review, which was expected to provide a referance for understanding and studing meatbolomics.

    Design and Construction of Problem-solving Agricultural Science and Technology Knowledge Service System
    WAN Min1,2, MENG Xian-xue1*
    2013, 15(2):  33-38.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.06
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    With facilitating of modernization construction of agriculture in China, farmers needs on agricultural information services has gradually changed to needs on professional and individualized science and technology (S&T) agricultural knowledge services, which aims at problem-resolving. Based on farmers needs, and aimed at the main issues of existing agricultural S&T knowledge service practices, the study develops a design and construction proposal of the problem-solving agricultural S&T knowledge service system for farm households. With “knowledge warehouse + service platform” and “diagnosis and service-push entity” being the core, the system strengthens information exchange and knowledge conversion among all components, through resources integration and mechanism innovation. The proposed system would provide convenient and efficient one-stop problem solving services for farmer householders, and would provide a new resolution for the agricultural S&T knowledge services as well.

    Agricultural Information Technology in Sichuan Province Based on SWOT Model Development Status Analysis and Counter Measures Studies
    TANG Jiang-yun, LI Xiao*, XIANG Ping, CAO Yan
    2013, 15(2):  39-44.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.07
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    Information technology in agriculture is an important means to change the traditional agriculture. It is an important symbol of a country or regions agricultural modernization and comprehensive strength. Facing intense domestic competition, Sichuan has to clear her own advantages and disadvantages in development of agricultural information technology, and try hard to synchronize with the advanced and spiry information technology. Only thus it can realize leap-forward development of agricultural economy. In this article, we used SWTO model to analyze agricultural information technology development in Sichuan under new situation. Sichuan agricultural information technology has advantages in infrastructure construction, service system and expert decision. It had main obstacles in precision agriculture and integrated personnel development. Before ending this paper put forward counter measures to promote healthy and sustainable development of Sichuan agricultural information technology.

    Analysis of Influential Factors on Grain Production in Gansu Province
    HAO Zhen-hua, YE De-ming*
    2013, 15(2):  45-53.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.08
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    It is of great significance for regional grain security to make clear the grain production capacity. Taking relevant grain production data from 1990 to 2011 in Gansu Province as sample, this article selected 9 indicators which had greater influence on grain production, and established a multiple linear regression model,and then analyzed the influential factors of grain production in Gansu Province by SPSS software. The results showed that those major influential factors were sowing areas of grain, applying quantity of chemical fertilizer, electric power consumption in rural areas, and areas affected by disasters. Their influence degree were 92.0%、24.8%、16.9%、-22.2%. According to the actual situation of Gansu Province, this paper put forward basic countermeasures to maintain the stability of grain production as following: in the premise of guaranteeing stable grain sowing areas, to further improve chemical fertilizer utilization efficiency; to strengthen power resources management and infrastructure construction of farm land, especially irrigation and water conservancy construction; and to speed up the innovation of agricultural science and technology.

    Analysis of Output and International Academic Influence of Academic Papers of Chinese Academy of Agricultural Sciences ——Based on Web of Science Database
    WANG Ting, YAN Yun, XU Yu-hong, MENG Xian-xue*
    2013, 15(2):  54-63.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.09
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    Empirical analysis and evaluation of scientific paper output and academic influence are not only the basic indicator to evaluate the level of research capability for agricultural research institutions, but also an important reference for decision-making of agricultural administrative department. In this paper, analysis was implemented through the amount of published papers, the distribution of partner countries or institutions, the cited frequency, the distribution of cited countries or institutions, and the subject structure of papers that published by Chinese Academy of Agricultural Sciences(CAAS) by using informetrics and scientific metrology. The results showed that the amount of SCI papers had grown rapidly since 2002 and development of research activities had shown a gradually active trend. The average annual volume of cited papers grew rapidly. It has been over 5.4 percent higher than the growth rate of paper output during the same period since 2004. The papers published by CAAS have being concerned by researchers from over 7 000 universities and research institutions, which were from more than 200 countries and regions. The international academic influence of CAAS is constantly expanding.

    Progress on Technology for Detection and Evaluation of Unintended Effects of Genetically Modified Plants
    ZHAO Jie, LIU Zhi, PENG Yu-fa*, QI Fang-jun*
    2013, 15(2):  64-69.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.10
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    Detection and evaluation of the unintended effects for genetically modified (GM) plants is one of the most important contents of safety assessment of GM plants. The definition, causes and progresses on the technologies for detecting and evaluating the unintended effects of GM plants were introduced. And the technical difficulties and existing problems about detection and evaluation of GM plants unintended effects were also pointed out. The potential application about the DNA microarray analysis technology used in detecting and evaluating the unintended effects of GM plants was specially emphasized. Besides, we had found out a non-uniform distribution pattern for differentially expressed genes of GM plants at different developmental stages and environments. This safety assessment mode may provide an experimental and theoretical basis for establishment of efficient, comprehensive, objective, impartial technologies, which can be applied to detecting and evaluating the unintended effects of GM plants.

    Ethylene and Plant Resistance to Adversity
    YU Yan-wen, HUANG Rong-feng*
    2013, 15(2):  70-75.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.11
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    Ethylene is an important plant hormone and plays important roles in plant tolerance to biotic and abiotic stress. This paper reviewed the recent research progress made on ethylene and plant resistance, the emphasis is introducing the significant role of ethylene in plant responses to drought, high-salt, low temperature, diseases, etc. stress, so as to provide basis for further understanding the regulating mode under adversity condition.

    VirA/VirG Two-component System Induced Infection of Agrobacterium tumefaciens
    YAO Yuan, YAO Yao, MO Ri-gen, FAN Li-fei*
    2013, 15(2):  76-82.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.12
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    Agrobacterium tumefaciens is a kind of gram-negative bacteria and present in soil. It can infect the wound parts of many varieties of dicotyledons, monocotyledons and gymnosperms, and induce the plant crown gall nodule by the product of vir genes located in the Ti plasmid. Thus, A. tumefaciens is an agricultural engineering bacterium, and has been widely used in plant genetic engineering. It has been shown that the expression of vir gene is regulated by VirA/VirG 2 component system. To summarize the molecular mechanism after A. tumefaciens infection, the paper expounds the structure and function of the VirA and VirG protein, and explains how the VirA/VirG two component system senses the environmental factors and how the expression of vir gene is regulated by these environmental factors through the VirA/VirG.

    Research of High Resistance to Turnip Mosaic Virus Mediated by Coat Protein Gene Segment
    ZENG Gang1,2, YE Yan-ying1, CAO Ming-qing1, MA Rong-cai1, TANG Le-chen2, YAO Lei
    2013, 15(2):  83-88.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.13
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    In order to get stable high resistance to turnip mosaic virus (TuMV), a 377-nucleotide (nt) segment near 3′ conserved region of TuMV coat protein (CP) gene was chosen to construct hairpin RNAi vector, and transformed into Arabidopsis thaliana. The transgenic plants were inoculated with TuMV isolate BJ-C4, which was of high pathogenicity and mainly epidemic in Beijing area. Three of the 8 transgenic lines showed hyper-resistance to TuMV. Quantitative PCR results revealed that the accumulation of virus was hardly to detect in highly resistant transgenic plants.

    Assessment of Genetic Diversity in Watermelon Based on SRAP Analysis
    LI Peng-fei, HUO Xiu-ai, CHENG Yong-qiang, DAI Liang, YANG Bing-yan, DUAN Hui-ju
    2013, 15(2):  89-96.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.14
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    Eighty watermelon materials were analyzed by SRAP (sequence-related amplified porlymorphism) technique. The results showed that 308 bands were amplified using 28 primer pairs, and 68 bands were polymorphic. Among the primer pairs, the minimum polymorphism frequency is 9.1%, the maximum is 45.5%, and the average is 22.1%. Based on DNA bands, the similarity coefficients were calculated and their value were ranged from 0.94~1.00, which indicated that their genetic base was narrow. Clustering analysis revealed that 80 watermelon materials were classified into 3 groups. The values of similarity coefficient among them were high. Materials from different districts were clustered together, which also showed that their genetic background was narrow. In order to get breakthrough and progress in watermelon breeding, we must strengthen the introduction and innovation of watermelon germplasm resources.

    Progress in the Study of Rhizobium Associated with Non-leguminous Plants
    LI Ting, HE Lai, LIANG Quan-feng*
    2013, 15(2):  97-102.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.15
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    In recent years, with the study progressing, and genetic engineering constantly matured, the scope of Rhizobiums host plant has been expanded and the growth-promoting effect on non-leguminous plants has been growing continuously. From the point of view of different growth-promoting mechanisms, this review summarizes the current knowledge and application on the Rhizobium associated with non-leguminous plants especially with gramineae.

    Gene Cloning and Characterization of a β-1,3(4)-Glucanase from Penicillium sp. C6
    DU Yan-long1,2, SHI Peng-jun2, ZHANG Xiu1*, YAO Bin2
    2013, 15(2):  103-109.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.16
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    Bglc6, a glycoside hydrolase with 16 families glucanase gene, was cloned from Penicillium sp. C6. The full-length of cDNA was 1 044 bp, including a putative signal peptide of 20 residues and a catalytic domain with glycosyl hydrolase of 16 families. The deduced amino acid sequence showed the highest identity (64%) with the putative endo-1,3(4)-β-glucanase from Grosmannia clavigera kw1407. The mature albumen BglC6 was successfully expressed in Pichia pastoris. The enzymatic characteristics analysis showed that the optimum pH and temperature for the recombinant glucanase BglC6 was pH 4.5 and 50℃. It exhibited good stability within acid pH range. When using barley β-glucan as substrate, the specific activity, Km and Vmax values were 253.8 U/mg, 8.42 mg/mL and 478.4 μmol/min·mg, respectively. BglC6 had better resistance to majority metal ions. Therefore, this glucanase possess potential application in animal feed industry.

    Research Progress of Tandem Repetitive Sequence in the Centromere of Plant
    HAO Wei-wei1,2, ZHOU Yan2*
    2013, 15(2):  110-114.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.17
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    Centromeres are the important domains of chromosomes that are responsible for sister chromatid cohesion, kinetochore assembly and spindle attachment, and are essential for proper chromosome segregation during mitosis and meiosis. Satellite DNA and retrotransposons are the most abundant DNA elements found in plant centromere regions. Centromeric tandem repeat play an important role in the centromere function and genome evolution. The study of centromeric tandem repeats got great progress for the development of sequencing technology. This paper introduces the development of centromeric tandem repeat of plants.

    Design of Monitoring System for Corn Ear Drying Based on Kingview
    ZHANG Da-long, ZHAO Wu-yun*, YANG Wan-xia, JIANG Wu-yang
    2013, 15(2):  115-119.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.18
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    In order to fully guarantee the seeds drying quality and germination rate, to avoid low seed drying efficiency and hand operation japse, we used Advantech ADAMN4000 series date acquisition modules, and designed a monitoring system for corn ear drying based on Kingview 6.53. Thus, the collection, treatment and monitoring of temperature and pressure data for corn ear drying was realized. Kingview was used to analyze, handle date and report fault alarm, so as to provide technical support to improve corn ear drying quality. Application shows that running of the control system is stable and reliable, man-machine interface is amicable. This system is able to meet the requirement of monitoring system for real-time and reliability.

    Studies on Harvesting Ship of Water Chestnuts Based on Soil Liquefaction Principle
    CHEN Hai-feng§, ZHANG Shi-xi§, YAN Yi-ye, CHEN Hao, XIE Qiang
    2013, 15(2):  120-124.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.19
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    Water chestnut grows in a special environment,and its harvest still relies on the traditional manual mining. The labor efficiency is low, and labor intensity is big. It is difficult to dig net. When as, it is difficult for potato harvest machinery to adapt to the rice field to harvest water chestnut. By analyzing the composition of soil where water chestnut grows in, a small experimental platform was built to carry out a test simulation of the mechanism of water chestnut digging according to the principle of soil liquefaction. A set of process suitable for rice paddy crop water chestnut mechanical mining and screening was put forward, and the theory of digging was explored. According to the analysis of experimental data, the main factors affecting water chestnut harvest have been obtained. The research showed that sandy loam can be liquefied by using the principle of soil liquefaction. The study provided a basis for the principle used for water chestnut harvest and provided design reference for water chestnut excavator prototype trial.

    Studies on the Rule of Airing Yak Beef Quality Forming and Microorganism Changing
    GAO Yuan1, YI Wen-zheng1, ZHANG Rong2, ZHOU Nan3, LIU Xuan1, SUN Bao-zhong1*
    2013, 15(2):  125-130.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.20
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    By detecting the quality and microorganism indicator of airing yak beef samples in different stages, we studied the rule of quality forming and changes of microorganism. The results showed that the appearance characteristics of the product change significantly and the weight loss rate was 76%; TBA value increased gradually in the processing process, and the final product was 0.48 mg/kg; the water content and water activity decreased gradually. Except the beginning day to the 5th day, change of moisture content was not significant (P>0.05), changes in the rest stages were significant (P<0.05). The 10thd to the 15th d was the major dehydration stage; water activity changes during the 20th d and 25th d were significant (P<0.05). In the airing period, the pH increased in a smaller changing range; change in the total number of colonies on the 5th d and 20th d were significant (P<0.05). The coliform reduced gradually in the process and no Listeria monocytogenes and Staphylococcus aureus were detected. In the drying process, the reduction of water activity was the main reason for the decline of total colonies number, and the lower water activity ensured the safety of products. The dehydration speed was the most important factor affecting air-dry duration and product crisp.

    Effect of Exogenous NO on H2O2 Metabolism in Postharvest Mango Fruit under Low Temperature Stress
    YANG Yang1,2,3, SHEN Lin2, SHENG Ji-ping2,4, WANG Feng-zhong1, WANG Feng1, FAN B
    2013, 15(2):  131-136.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.21
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    Nitric oxide (NO) is an important signal molecule in plants. In order to discuss the effect of NO on H2O2 level and metabolism in postharvest mango fruit under low temperature stress, 0.08 mmol/L NO donor SNP and 30 μL/L NO gas were used to treat green mango fruit before cold storage. H2O2 content and the activities of superoxide dismutase (SOD), catalase (CAT), peroxidase(POD) and ascorbate peroxidase (APX) were measured 12 d after cold storage at 3℃. The results showed that both SNP and NO gas treatments had significantly reduced H2O2 level in postharvest mango fruit pericarp, and increased the activities of SOD, CAT and POD. The effect of SNP was more significant than NO gas. The main effect site of exogenous NO was pericarp.

    Research Progress on Disinfection Technology of Wastewater from Animal Production
    LI Tong, DONG Hong-min*, TAO Xiu-ping
    2013, 15(2):  137-143.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.22
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    Livestock wastewater reuse is one of the essential measures to control environment pollution from the livestock production of our country in the future. Disinfection is critical to ensure the safety of wastewater reuse. The mechanism of disinfection technology including chlorine dioxide, ozone, electrolyzed water, ultraviolet, and ultrasonic for wastewater reuse, technical feature, and application status were reviewed in this paper. Further exploration of appropriate technology for livestock wastewater disinfection will provide theoretical basis for research and application of livestock wastewater disinfection and reuse technology.

    Studies on Sink-source Characteristics of Mid-season Hybrid Rice for High Yield in Winter Water Logged Field
    XU Fu-xian1, YAN Yun-de2, XIONG Hong1, ZHANG Lin1, GUO Xiao-yi1, ZHU Yong-chuan1
    2013, 15(2):  144-152.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.23
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    Taking 30 mid-season hybrid rice combinations as material, the relationship between the grain yield and the sink-source characteristics of hybrid rice were researched in 3 types of winter water logged field (drought field in summer, ordinary field and store paddy), respectively. The results indicated that the yield of mid-season hybrid rice was greatly influenced by the types of paddy field. There was the same variation trend of grain yield for 30 mid-season hybrid rice combinations in 3 different types of paddy field. 7 hybrid rice combinations could get high yield in 3 types of winter water logged field. The high-yield goals in winter water logged field were to further increase seed setting percentage and 1 000-grain weight, on the basis of securing bigger number of spikelets per unit area. There were parabolas relationship between the theoratical grain yield and production potential, and spikelets per panicle, sink plumpness index, respectively. The high-yield combinations for winter water logged field had the following characteristics: 160~200 spikelets per panicle, maximum tillers 332.98×104~360.98×104 per hm2, productive panicles 200.14×104~225.24×104 per hm2, seed setting percentage 77.56%~82.38%, 1000-grain weight 2906~30.32 g, spikelets number 3.61×104~3.98×104 per/m2, leaf area index 5.69~6.59, spikelet number to leaf area ratio 0.55~0.70 spikelet/cm2, single ear weight 4.00~4.51 g. These results provide a scientific basis for rice breeders to identify high yield combination, and production sections to screen rice varieties suitable to grow in different types of paddy fields in winter water logged field areas.

    Research Progress on Transfer and Distribution of Heavy Metal in Tobacco and Cigarette
    LIU Hai-wei, SHI Yi*, LIANG Hong-bo
    2013, 15(2):  153-158.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.24
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    Heavy metal damage to environment has drew wide attention of the public in recent years. The security of tobacco products has also been closely concerned by the people. This article summarized the following 3 aspects: sources of heavy metal in tobacco products, the distribution and accumulation of heavy metal in different organs of tobacco, and the transfer of heavy metal in cigarette and smoke. It pointed out the key links of controlling heavy metal are absorption, distribution, procession, filtration and absorbability by human body. This article also gave some suggestions about the control methods of heavy metal in tobacco products, the differences between regions and breeds, the mechanisms and the affecting factors of absorption and distribution of heavy metal in tobacco plant, and the formulation of standards for limited quantity of heavy metal in tobacco products, which was excepted to provide reference for heavy metal control.

    Full-length cDNA Cloning and Expression Analysis of Profilin Gene from Sea Cucumber (Apostichopus japonicus)
    DONG Ying, GAO Shan, CHEN Zhong, YANG Ai-fu, JIANG Bei, GUAN Xiao-yan,
    2013, 15(2):  159-167.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.25
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    Sea cucumber (Apostichopus japonicus) was one of the important aquaculture species in the north coast of China. The studies on function and mechanism of immune-related genes can provide a reference to the prevention of diseases in the cultured A. japonicus. In this study,a full-length cDNA sequence of profilin gene from A. japonicus was obtained by constructing cDNA libraries and sequencing analysis, which was 787 bp and includes a 205 bp 5′- untranslated region (UTR), 378 bp ORF encoding 125 amino acids with a conserved PROF domain, and a 204 bp 3′-UTR; and the predicted molecular weight was 13.4 kDa. The conserved PROF domain included acting interaction sites, poly-L-proline binding sites, and PIP2-interaction sites. The deduced amino acid sequence showed 46% amino acid sequence identity to the Profilin of Trichoplax adhaerens and Saccoglossus kowalevskii. The profilin mRNA levels were detected by using quantitative real-time PCR at different developmental stages of sea cucumber, including unfertilized egg, fertilized egg, cellulous stages, blastula, gastrula, early auricularia, auricularia, late auricularia, doliolaria, pentactula and juvenile. The expression patterns were also detected in different tissues of young sea cucumbers including intestines, coelomocytes, respiratory trees and body walls. The profilin expression levels were low from unfertilized eggs to gastrula, however, an increase in transcript levels was observed from early auricularia to juvenile. The highest expression level was found in the coelomocytes of young sea cucumbers. LPS was a potent activator of the echinoderm immune system,and after LPS challenge, the highest expression of profilin mRNA was observed in the coelomocytes.

    PCR Detection Method of Shewanella aquimarina
    WANG Yi-nan, MU Xiao-hu, FENG Ni-sha, DING Jun, CHANG Ya-qing*
    2013, 15(2):  168-172.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.26
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    Shewanella aquimarina is one of the pathogenic bacteria of sea urchin Strongylocentrotus intermedius. To establish PCR detection process of S. aquimarina, 3 pairs of primer were designed according to the specified region of gyrB genes of S. aquimarina, and the specificity and sensitivity were analyzed. The results showed that the amplified fragments of PCR applied with primer SA-9 was agreed with the expected sizes. SA-9 could detect the minimum bacteria concentration of 4.6×103 cfu/mL, and the minimum detectable concentration of bacteria DNA was 0.315 pg/μL. These experiments showed that primer SA-9 had high specificity and sensitivity to S. aquimarina. S. aquimarina in coelomic fluid of sea urchins and the environmental samples were detected by the method. The results of healthy sea urchins, breeding seawater and feeding kelp were negative, while the sample of seawater from natural sea area had a certain amount of S. aquimarina.

    Comparative Analysis on Microchip Electrophoresis Technology and Polyacrylamide Gel Electrophoresis
    ZHANG Sha-sha1, GUO Xin-mei2, ZHAO Mei-ai1, PEI Yu-he2, SONG Xi-yun2*
    2013, 15(2):  173-178.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.27
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    Polyacrylamide gel electrophoresis (PAGE) is usually applied in separation of oligonucleotides. Microchip electrophoresis is a new electrophoretic technology to detect the molecular size of DNA and RNA samples. In order to identify the repeatability and precision of microchip electrophoresis, PCR products of Black Glutinous Maize and self intersect line 8112 etc. were tested using PAGE and microchip electrophoresis, respectively. The results showed that compared with PAGE, the latter had no toxin, and could be operated easily, with high sensitivity and accuracy, and had better repeatability. So it might present broader prospects in scientific research in future.

    Studies on Fermentation of Bacillus mucilaginosus WY120 from Corn Stalk Enzymatic Hydrolysate
    DU Ying1,2, WANG Zi-qiang2,3, XU Xiao-juan2,3, ZHANG Li-ping1, WANG Yun-shan2*,
    2013, 15(2):  179-184.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.28
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    Bacillus mucilaginosus is an usual strain of microbial fertilizer. Main raw materials such as starch, sucrose, glucose, etc. are used in traditional fermentation of biological potassium fertilizer. With corn stalk as raw material, this paper compared fermentation of Bacillus mucilaginosus in starch and sucrose medium and corn stalk enzymatic hydrolysate medium; discussed the feasibility of taking corn stalk enzymatic hydrolysate as carbon source. The results showed that the viable number of 3.52×108 cfu/mL was obtained using corn stalk enzymatic hydrolysate as carbon source, which was 1.63 times of the starch and sucrose culture medium. The result of optimization experiment of corn stalk enzymatic hydrolysate medium showed that the best glucose concentration was 15 g/L and the viable number reached 3.95×108 cfu/mL. The result of 5 L fermenter experiment indicated that viable number reached 6.76×108 cfu/mL under the condition of rotation speed 250 r/min and pH 7.0~7.2, which was 1.71 times of shake flask culture. The results indicated that it was possible that instead of starch or sucrose, corn stalk hydrolysate can ferments Bacillus mucilaginosus, which provides a new way for utilization of corn stalk.

    Medium Optimization for Phosphate-solubilizing Bacteria Bacillus Megaterium in Submerged Fermentation
    WANG Jin-ling1, LIU Xiao-ping2, GAO Wei-hua1, ZHAO Feng-yan3, LV Chang-shan3
    2013, 15(2):  185-192.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.29
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    Response surface optimized experiment was carried out to gain a best culture medium with high density zymic. This experiment tool a strain of phosphate-solubilizing bacteria Bacillus megaterium isolated from soil as research objective, and OD600 of the broth as a standard to judge the number of viable cells. Firstly, single element experiment was conducted to screen the carbon source, nitrogen source, carbon source concentration, nitrogen source concentration, inorganic salts and its concentration from the medium, and achieve the best value of single element. Then 3 steps experiments namely fractional factorial design (FFD), steepest ascent experiment and central composite design (CCD) were conducted to optimize the culture medium by using response surface design. The results showed that the optimal culture medium composition included: lactose 6 g/L, peptone 7.45 g/L, NaCl of 5 g/L, MgSO4·7H2O 2.46 g/L, CaCl2 1.22 g/L, and K2SO4 0.087 g/L. Under this optimal culture medium, the number of viable cells of Bacillus megaterium in ferment fluid could reach over 2.0×109 cfu/mL. Fermentation of medium by this formulation could provide high density phosphate-solubilizing bacterial fertilizer for agriculture production.

    Progress on Ligation-independent PCR-mediated Approaches for Genomic Walking
    LI Xu-juan1, CHEN Yang-ling1, WANG Hai-bo1,2, GONG Ming1, ZOU Zhu-rong1*
    2013, 15(2):  193-199.  DOI: doi:10.3969/j.issn.1008-0864.2013.02.30
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    PCR-mediated approaches for genomic walking (or chromosome walking) are routinely used for cloning flanking sequences of the known DNA regions, which is an important technology for genomic research. According to the principles, they can be categorized into two types of ligation-dependence and ligation-independence. At present, numerous approaches of the later, each with distinct features, have been rapidly developed. Nevertheless, regarding the binding patterns of walking primers on DNA templates, they can be overall grouped into the degenerate primer-based or specific sequence-dependent subsets. Following this viewpoint, this paper summarized the major or representative techniques and the recent developments of ligation-independent, PCR-mediated genomic walking, so as to provide a methodological reference for the relevant researches.