Table of Content

15 June 2013, Volume 15 Issue 3

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Development Process, Problem and Countermeasure of Maize Production in China

ZHAO Jiu-ran, WANG Rong-huan

2013, 15(3):  1-6.  DOI: 10.3969/j.issn.1008-0864.2013.03.01

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Maize has become the 1st major crop with the largest planting area and total yield in China, and it is strategically significant to promote the maize production for ensuring the national food security. In this paper, the development process of maize production in China since the reform and opening up and the typical features for each phases were introduced systematically, and the main problems and factors limiting the maize production such as the distribution of natural resource, agronomic infrastructure, maize variety and seed quality, cultivation technology, the production and management mode, the storage and circulation conditions were also analyzed. At last, some policy suggestions including increasing and strengthening the scientific research input, basic infrastructure, supporting policy, technical services, high-yield construction, storage and circulation conditions, and technical measures such as maize new variety breeding, water-saving agricultural technology, conservation tillage, improving the sowing quality and population uniformity, mechanized cultivation, diseases & insects and weeds protection etc. for the further increasing of maize yield level in the future were purposed, respectively.

Research Progress on Structure and Function of Maize Transcription Factors

MO Xiao-ting, ZHAO Jun*, FAN Yun-liu, WANG Lei*

2013, 15(3):  7-17.  DOI: 10.3969/j.issn.1008-0864.2013.03.02

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Transcription factors play important roles in controlling diverse processes of plant growth and development, and responses to environment. So far studies on transcription factors are mainly focased on Arabidopsis and rice, while the structure and function of maize transcription factors are less studied. According to the Plant TFDB database, maize transcription factors are divided into 56 families. We summarized the classification of maize transcription factors and their research progress on structure and function in plants, aming to promote the relevant studies on maize transcription factors.

Progress on Major Gene/QTL for Disease Resistance in Maize

XU Ling§, ZUO Wei-liang§, LIU Yong-jie§, LIU Qing-qing§, TAO Yong-fu§,

2013, 15(3):  18-29.  DOI: 10.3969/j.issn.1008-0864.2013.03.03

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A wide range of pathogenic microorganisms, including fungi, bacteria, viruses and oomycetes, induce diseases on leaf, florescence, stalk and root of maize, and pose a grave threat on maize production. Maize is a cross-pollinated crop and rich in genetic resource. Maize has highly diverse in genome architecture and contains a huge number of qualitative and quantitative resistance loci. During the past few years, a great progress has been made towards the gene-mapping and exploration of major disease resistance gene/QTL in maize. Until now, at least 4 major resistance genes were cloned and a number of candidate genes were obtained, along with large-scale mapping of numerous resistance gene/QTL. These researches greatly accelerated marker-assisted introgression or polymerization of resistance genes/QTLs into high-yield potential but susceptible maize lines, and this is proved to be very efficiency in resistance breeding program. In this paper, we summarized the recent progress on maize resistance to major diseases caused by fungi, viruses, bacteria and oomycetes.

Improvement of Maize Hybrids and the Analysis of Basal Germplasm in China

LI Yong-xiang, SHI Yun-su, SONG Yan-chun, LI Yu| WANG Tian-yu

2013, 15(3):  30-35.  DOI: 10.3969/j.issn.1008-0864.2013.03.04

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Improvement of maize hybrids was one of the key factors for the continuous increasing of

corn production in China. Under the premise of durative decreasing of cultivated area, the

increasing of yield potential by breeding would be crucial for maize production. In this

paper, we summarized the contributions of variety improvement to the increasing of yield per

unit area in China, the characters of improvement of Chinese maize hybrids and future

requirement, as well as the enhancement and status quo of breeding germplasm and potential

development in China. Also, we discussed the high density and high nutrient-efficiency

breeding strategy in China, the method of germplasm enhancement, and establishment of high

efficiency germplasm evaluation & enhancement platform, which were exprected to provide some

basic information for maize breeding.

Process on Glyphosate-resistant Transgenic Maize

XIE Shu-zhang1, YANG Xiao-yan1, LIN Qing1, WENG Jian-feng2, JIANG Xiao-ying1, LI

2013, 15(3):  36-41.  DOI: 10.3969/j.issn.1008-0864.2013.03.05

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Glyphosate is a broad-spectrum, inner-absorption conducting herbicide that has been widely used in maize production. With development of plant genetic engineering technology and continuously expansion of genetically modified crops planting area, the research and development of glyphosate-resistant transgenic maize have become an important part of maize breeding. In this paper, we introduced the function mechanism of glyphosate, analyzed in detail the strategy of glyphosate-resistant plant genetic engineering. The paper also systematically reviewed the internal and external progress made in research and development of glyphosate-resistant transgenic maize, and its commercial application. It put forward strategy for studying and developing glyphosate-resistant transgenic maize in China, so as to provide relevant references for enhancing glyphosate-resistant transgenic maize research in China.

Preliminary Studies on Corn Commercial Breeding and Seed Industrialization in China

YANG Jin-sheng1, WANG Gui1, BAI Na2, LI Xu-hua1, JIA Chun-lan1,

2013, 15(3):  42-47.  DOI: 10.3969/j.issn.1008-0864.2013.03.06

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In 2012, corn production in China surpassed rice and became the No. 1 grain crop. Corn plays an increasingly important role in grain production and national economy. Exploring the establishment of a new system for commercialization of corn breeding and industrialization of seed production has become an important issue facing Chinas corn seed industry. This paper compared the seed industry development at home and abroad, analyzed the existing problems in the commercial corn breeding and gaps between China and other countries. And pointed out the opportunities, challenges and development trend for Chinas seed production industry.

Synthetic Research and Utilization on Industrial Development of Barley in Southwestern China

ZENG Ya-wen1*, PU Xiao-ying1, ZHANG Jing2, GUO Gang-gang2, DU Juan1, YANG Tao1,

2013, 15(3):  48-56.  DOI: 10.3969/j.issn.1008-0864.2013.03.07

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In order to promote the rapid development of barley industry in Southwestern China and solve the problems of declining barley production in China, this paper summarized the successful experience in coordinating the barley production and biodiversity, malt barley and beer industry, feed barley and animal husbandry, functional barley and functional food, and existing problems facing these four industries. This paper put forward the thoughts and countermeasures for sustainable and stable development of barley industry, especially put forward for the first time that human chronic disease outbreak was rooted from taking brown rice and barley as major food in ancient time into taking white rice and refined wheat flour as staple food in modern days. These research results had provided scientific basis for the sustainable development of barley industry in China, and also supported the decision made by the government.

Planting Mode of Modern Genuine Traditional Chinese Medicinal Material and its Production Base Innovation Exploration

SUN Jun-she1, ZHENG Zhi-an2, ZHANG Xiu-qing2, LUO Jiang-qing3, YAO Song-jun3, ZH

2013, 15(3):  57-63.  DOI: 10.3969/j.issn.1008-0864.2013.03.08

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Under the background of modern agriculture, the planting and management mode of genuine traditional Chinese medicinal material is the headstone for developing large scale traditional Chinese Medicine industry in China. From the perspective of modern agriculture, this paper discussed the innovation both in the modes of planting genuine traditional Chinese medicinal materials and co-building of their planting bases. Based on the planting characteristics, a new intensive and industrialized planting mode was explored, which needs the innovations in technology, organization, mechanism and procedure, and closed management and risk sharing would be realized. Based on the theory of cooperative game, a new mode suitable for planting base cooperation was constructed, i.e. 2 platforms, 3 mechanisms and 4 communities. At last Zhejiang Longquan Keda Agricultural Byproducts Co. Ltd. tested and verified the implementation effect.

Research and Development of Livestock Reproductive Biotechnology and its Industrialization and Popularization

LI Xi-he1,2

2013, 15(3):  64-71.  DOI: 10.3969/j.issn.1008-0864.2013.03.09

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Live stock reproductive biotechnology includes artificial insemination,in vitro fertilization, embryo transfer, animal sex control, transgenic animal, animal cloning and stem cell technology etc. Early studies on reproductive biotechnology mainly began in the twenty century and has experienced about 100 years from study to industrialization, extension and application. At present, this technology has been widely applied both in live stock industry, including animal cell sex control, gene transfer-Colone technology and stem cell technology, and in human medical treatment.

Needs Analysis of Problem-solving Agricultural Scientific and Technological Knowledge Service

WAN Min1,2, MENG Xian-xue1*

2013, 15(3):  72-76.  DOI: 10.3969/j.issn.1008-0864.2013.03.10

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Farmers needs analysis is the groundwork for agricultural science and technology (S&T) knowledge service. This paper reviewed the history of farmers needs research on information and knowledge in China. In view of the differences between knowledge service and traditional information service, this paper put forward the thoughts of taking the farmer households difficult problems in agricultural production and management as starting point to study their needs for knowledge service, and also analyzed the most concerned problems by the traditional small farmer householders, specialized farming / animal husbandry householders, and farmer specialized cooperation during agriculture production and management. So the paper explored new research thoughts and lays solid foundation for the studies on problem-solving agricultural S&T knowledge service system.

Studies on Biological Characterization of Rice Brittle Culm Mutants and Their Biomass Degradation Efficiency

FENG Yong-qing1, ZOU Wei-hua1, LI Feng-cheng1, ZHANG Jing1, ZHANG Hui1,

2013, 15(3):  77-83.  DOI: 10.3969/j.issn.1008-0864.2013.03.11

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Brittle culm mutants are considered as valuable germplasm resources for energy crop breeding in rice. This study analyzed biological characters and biomass digestibilities of 10 distinct rice brittle culm mutants. Compared with the wild type japonical variety NPB (Nipponbare), all mutants displayed normal growth and development, and shorter plant height and lower tension stress. In terms of cell wall composition in the mature culm tissues, the mutants showed lower cellulose and higher hemicelluloses levels than that of the NPB and no significant increase is identified in lignin level, which were consistent with the RNA transcript levels of their major wall polymer synthases. Due to cell wall alternation, the rice mutants appeared to have a stronger resistance to lodging than NPB did. However, all mutants and NPB were determined with unchanged single plant spike weight and higher number of effective panicles, suggesting that the mutants were not much affected with their seed yield. Meanwhile, the mutants had higher biomass yield. With respect to the brittle characters, all mutants exhibited a much higher biomass enzymatic digestibility upon various acid and alkali pretreatments. The data indicated that the mutants could be applied for energy crop breeding towards high yields of seeds and biomass in rice.

Cloning and Expression Pattern Analysis of Gossypium hirsutum Lipid Transfer Protein Gene Family

HAN Hui-chao1, HUANG Yi-qun1, WANG Jin2, ZUO Kai-jing1*

2013, 15(3):  84-90.  DOI: 10.3969/j.issn.1008-0864.2013.03.12

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Long-chain fatty acids could activate cotton fiber initiation and elongation by regulating intracellular ethylene signal transduction in cotton ovule. In order to fully understand how fatty acids are transferred to the surface of fiber cells and how many genes are involved in this process, we cloned 11 LTP genes after genome-wide search and analyzed their expression patterns during fibers development by quantitative RT-PCR. The results showed that there are differences between expressions of LTP6, LTP7, LTP9 and LTP11, and these 4 genes might be closely related to cotton fiber development at the original stage. We also cloned Gh-LTP6 promoter by Genome Walking technique and analyzed its motif, results showed there are mutiple MYB binding site. These results might lay a solid foundation for intensive studying the role of LTP genes in the process of cotton fiber development.

Construction of High-performance Bivalent (Bt+cpti) Expression Vector and its Expression Analysis

XUE Ji-xiong1,2, ZHANG Rui1, ZHANG Hua2, SUN Guo-qing1,

2013, 15(3):  91-97.  DOI: 10.3969/j.issn.1008-0864.2013.03.13

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In this study, the Bt and cpti gene were modified on the base of existing vector and expression vector. Soybean trypsin inhibitor SKTI signal peptide sequence was added at 5′ end of cpti, KDEL signal peptide sequence was added at its 3′ end, and at Bt 5′ end chloroplast targeting peptides was added, and then a bivalent expression vector PGBIC(K).B.4A with signal peptides and a control vector PGBIC.B.4A without signal peptides were constructed. Through pollen tube pathway mediated by agro-bacterium Gossypium hirsutum Y18 was transformed. The result of southern blot showed that the exogenous gene had been integrated into the receptor with genome different copies. By ELISA, it was found that the protein expression was increase by 1~8 times in 5 positive strains. In the insecticidal tests, the modified binary vector transgenic plants1/4-1showed higher resistance to the bollworm because of the accumulation of protein. This study provided a new method to obtain the higher resistance of bivalent transgenic cotton, and basis to improve the accumulation of other foreign protein in genetic engineering.

Initial Exploration of Tomato RING BOX Protein 1 (RBX1) Gene Function

TIAN Xue-fen, TANG Xiao-feng, LI Di, LIU Yong-sheng*

2013, 15(3):  98-107.  DOI: 10.3969/j.issn.1008-0864.2013.03.14

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E3 ubiquitin ligases are very important regulatory components for the ubiquitin proteasome pathway. Most of E3 ligases comprise multiple groups and contain scaffolding proteins known as cullins and a common catalytic subunit, RBX1. RBX1 has a function to bind the ubiquitin-conjugating enzyme E2 and makes it into close proximity with the Cullin-based E3 substrate. In this study, yeast two-hybrid showed the tomato RBX1 can be interacted with CUL4 directly. We examined the expression level of RBX1 genes in different tomato tissues of wild type, and the result showed that RBX1 gene was expressed constitutively, while higher expression level was detected in the flower and fruit, comparing with that of leaf and root. In addition, an over-expression vector composing of aimed gene fragment and PBI121 with CaMV35S promoter, named PBI121-35S-RBX1 was constructed. We introduced the constructed vector into tomato cv. Ailsa Craig via Agrobacterium-mediated transformation, and acquired 3 transgenic plants of co-suppression and 3 transgenic plants of over-expressing, respectively. An increase in leaf expansion, a reduction in apical dominance comparing with wild type and sterility were observed in transgenic plants of co-suppression, which indicated that RBX1 played an important role in plant growth and development. We speculated that plant RBX1 gene may involve with auxin and brassinosteroid response and proliferation of cells, which provided clues for further studies on RBX1 in plants.

Research Progress of microRNA on Animal Phenotypic Regulation

WANG Chen, ZHAO Wei-min, MOU Yu-lian*

2013, 15(3):  108-112.  DOI: 10.3969/j.issn.1008-0864.2013.03.15

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microRNA (miRNA) is a class of endogenous gene which encoding non-coding single stranded RNA molecules with length about 21~22 nucleotides. They regulate the expression of target genes through gene silencing. With the further research of large-scale transcriptome, a large number of microRNAs were found be related to animal phenotypes. This article briefly introduced the basic features of microRNAs, and summarized the rescent progress on determining traits and in animal. A growing number of miRNA is associated with a variety of animals important phenotypes. It probably provided a new way for studying of Animal phenotypic regulation.

Progress on General Nitrogen Regulation Protein NtrC

2013, 15(3):  113-122.  DOI: 10.3969/j.issn.1008-0864.2013.03.16

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NtrBC is a two-component system involved in bacteria nitrogen metabolism regulation, and NtrC is a transcription activator protein in response to environmental signal and activates the transcription of target genes. The past studies have indicated that bacterial NtrC plays important roles in regulating diverse processes including nitrogenous compounds utilization, biological nitrogen fixation, the biosynthesis of biopolymer and maintenance on the carbon-nitrogen balance. Studies on NtrCs overall regulating function are one of the hotspot of microbe metabolic regulatory networks. This paper reviewed the research progress on NtrC in recent years, and described in detail the biological functions of bacterial NtrC, so as to better understand microbian environment adaptation, metabolic diversity, and the molecular mechanism of C-N coupling regulation.

The Effect of AmrB on Aeromonas hydrophila Aminoglycosides Resistance

ZHANG Dan-feng1, CHEN Guo-ping2, ZHANG Guo-guang1, HUANG Jia-fu1, HUA Xiu-ting1,

2013, 15(3):  123-128.  DOI: 10.3969/j.issn.1008-0864.2013.03.17

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AmrB is a crucial member of drug efflux pump. Investigating the relationship between AmrB and the aminoglycosides resistance of Aeromonas hydrophila is supposed to be important for understanding the mechanism of its drug resistance. amrB gene was cloned to pET-28a vector and confirmed by double digestion and sequencing, thus obtained amrB-pET-28a. amrB-pET-28a/BL21 was induced by IPTG to obtain AmrB protein then purified by Ni resin. New Zealand rabbit was immunized by AmrB to get AmrB serum with a titer of 1∶6 400. Kanamycin, streptomycin and gentamycin resistant strains of Aeromonas hydrophila AH1 were screened by 10 serial culture with 1/2 MIC of corresponding antibiotic. Western blotting was performed to analyze the expression of AmrB in drug resistant strains. After analysis of Western blotting by Phoretix 1D software and SPSS software, the expression of AmrB in kanamycin, streptomycin and gentamycin resistant strains was significantly 2.16,2.65 and 2.13 folds higher than control strain, respectively. AmrB plays an important role in the aminoglycosides resistance of Aeromonas hydrophila AH1, which may offer an opportunity to further explore the molecular mechanisms within.

Studies on Service System of Intelligent Mobile Phone Based on Agriculture

WU Jian-wei1, LIU Wei2, LU Bing-you3, YANG Bao-zhu1,2*

2013, 15(3):  129-134.  DOI: 10.3969/j.issn.1008-0864.2013.03.18

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Agricultural intelligent mobile phone is an equipment, which can provide portable, simple operation with rich professional information for modern agricultural development. With modern agricultural production management as main object, combined with the development status in mobile phone industry, we studied thoroughly the  embedded technology and intelligent decision technology, etc. key technologies, including collection, processing and integration 25 series of agricultural knowledge of crop, vegetable, fruit tree, flowers, animal husbandry, aquatic product 6 aspects. We also studied on corn, wheat, cotton, cucumber, peach, grape, pig, dairy cow, China rose, lawn, freshwater fish, etc., so as to realize data collection and management, intelligent decision, SMS push, voice prompts, 3G video services and other functions, thus to effectively accelerate the pace for innovation of agricultural informationization of science and technology.

Regulation Responses on Rice Root Formation and Elongation by Low pH and Aluminum Stress in South China

FU Xue-lin, CHEN Wei-dong, ZHONG Sheng-zhi, ZENG Huan-huan,

2013, 15(3):  135-142.  DOI: 10.3969/j.issn.1008-0864.2013.03.19

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Soil in southern China is acid and red, P deficiency and Al toxicity commonly coexist on this kind of soil. It is one of the important limiting factors on rice production in southern China. In order to elucidate Al-tolerance of Yuexiangzhan, an elite indica cultivar which was widely planted in South China, the experiment of aluminum and pH on root formation and growth in germination and seedling stage was undertaken in simple CaCl2 solution by using Al-tolerant japonica cultivar, Nipponbare, as a control. In the process of seed germination, Al inhibited seed root formation and growth significantly (F=123.84,P<0.000 1). However, the differences in sensitivity to Al stress at the stage of primary root formation between Yuexiangzhan and Nipponbare were not significant. Seedling root elongation was affected by solution pH both in Nipponbare and Yuexiangzhan after 24 h cultured with 0.5 cm root in 0.5 mmol/L CaCl2 solution with different pH (pH 3.5, 4.5, 5.5, 6.5, 7.5). Root length (RE) was significantly affected by low pH value of the solution (F=70.01,P<0.000 1). The lower the solution pH value was, the shorter the seedling roots would be. In dose-response experiment, Al tolerance of the two cultivars was significantly different (F=25.71, P=0.000 2), Nipponbare showed higher tolerance at various Al concentrations than Yuexiangzhan. 50 μmol/L Al was thought to be the key concentration to discriminate tolerance of rice genotypes to Al stress in simple CaCl2 solutions. In time-course experiment, control root elongation (CRE) and stress root elongation (SRE) all increased along with the culture time prolonged from 6 h to 36 h. But differences of relative root elongation (RRE) between cultivars were significant (F=21.79, P<0.000 1), also in Al stress times (F=5.38,P=0001 3). RRE of Nipponbare was significantly higher than that of Yuexiangzhan. Analysis results of Al content in root tips or apexes were consistent in qualitatively analysis by hematoxylin staining, and quantitatively determination by method of graphite furnace atomic absorption spectrophotometer. There is an increasing trend of Al content in seedling root tips with the increasing of Al concentrations in culture solutions, while more Al was accumulated in seedling root tips of Yuexiangzhan than that in Nipponbare. Based on these results Yuexiangzhan was considered as an Al-sensitive cultivar.

Effects of Banning Grazing and Delaying Grazing on Species Diversity and Biomass of Alpine Meadow in Northern Tibet

ZHANG Wei-na1, Ganjurjav1, LI Ya-wei2, GAO Qing-zhu1*, WAN Yun-fan1,

2013, 15(3):  143-149.  DOI: 10.3969/j.issn.1008-0864.2013.03.20

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Banning grazing and delaying grazing are reasonable grassland management measures for restoring the degraded alpine grassland. In this study, 5 treatment plots including banning grazing for 3 years (JM3), for 5 years (JM5), for 7 years (JM7), delaying grazing for 5 years (XM5) and continuous grazing (FM) plots were devised in Nagqu to investigate the effects of banning grazing and delaying grazing on alpine meadow in Northern Tibet. Based on the quadrant method, community species diversity, height, coverage, aboveground biomass and underground biomass within areas of 0.5 m×0.5 m quadrant of alpine meadow in Northern Tibet were recorded. The result showed that the richness index and diversity index in banning grazing plots were significantly higher than those in XM5 and FM. JM3 and JM5 were significantly higher than JM7. But the evenness index in JM3 was significantly lower than that in JM5 and JM7. The important values of the grasses and sedges in JM3 were higher than other plots. The aboveground biomass of JM5 was 84.2 g/m2 and higher than other plots. The ratio of underground biomass and aboveground biomass of JM5 was the least in all plots. Thus, binning grazing for 5 years could maintain higher species diversity and significantly increase the productivity of alpine meadow, but binning grazing for over 5 years would go against maintaining higher species diversity and meadow available biomass.

Research Status and Prospects of Megastigmatrienone in Flue-cured Tobacco

WU Yan-hui1, XUE Li-xin2, XU Zi-cheng1, ZHU Jin-feng3, XIE Xiao-hui2,

2013, 15(3):  150-156.  DOI: 10.3969/j.issn.1008-0864.2013.03.21

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Megastigmatrienone is an important natural aroma component of flue-cured tobacco, which has a great effect on tobacco flavor and taste. The formation and gene regulation of megastigmatrienone in tobacco leaves were summarized based on the function of megastigmatrienone. And the influences of genotype, ecological factors and cultivation measures on its content were expounded. This paper suggested that more emphasis should be put on studying the influences of various factors on syhthesis enzyme of megastigmatrienone, and put forth effort on mousing out the ordor recceptor of megastigmatrienone in nasal cavity, so as to improve tobacco quality purposefully, and provide theoretical basis for developing flue-cured tobacco of superior quality.

Studies on Antioxidant Activity of Products from Fermentation of Hypsizigus marmoreus in Kelp Waste

ZHANG Bing-zhao1, YAN Pei-sheng1,2,*

2013, 15(3):  157-162.  DOI: 10.3969/j.issn.1008-0864.2013.03.22

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In order to develop a new pathway to utilize kelp waste, Hypsizigus marmoreus was incubated in medium mainly composed of kelp waste and products from fermentation were investigated with antioxidant assays. The results showed that mushroom could grow in this medium and produce abundant mycelia. Antioxidant assays indicated that the total antioxidant capacity (T-AOC) of the supernatant and extract of mycelia were 0.23±0.02 U/mg and 0.77±0.01 U/mg, which were 43.8% and 18.5% higher than those of the medium; and also significantly higher than that of the chemical antioxidant BHT. Furthermore, the supernatant of fermentation broth exhibited potent capacity for hydroxyl radical scavenging and superoxide anion radical scavenging, as their EC50 values were 0.43 mg/mL and 26.02 mg/mL. The study showed that the compounds with potent antioxidant activity produced during the fermentation mainly distribute inside of the mycelia, and the compounds with potent capacity for hydroxyl radical scavenging mainly distribute in the supernatant.

Effect of Salinity Stress on Immune Enzyme Activity of Sea Cucumber (Apostichopus japonicus)

WANG Chong1, TIAN Yi2*, CHANG Ya-qing2, CHEN Bai-yao3,4,

2013, 15(3):  163-168.  DOI: 10.3969/j.issn.1008-0864.2013.03.23

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The behavior of sea cummber and the activities of enzyme would change under salt stress to adapt the environment. The sea cucumbers were temporarily cultured at different salinity levels 18, 23, 33, 36 and 40. The enzyme activity of alkaline phosphatase (AKP), acid phosphatase (ACP), lysozyme (LSZ) and superoxide dismutase (SOD) of sea cucumber were detected under different salinity stress. Survival was examined after the larvae were exposed to the salinity levels. The results indicated that salinity stress could affect sea cucumber activity and the lower salinity had more effect than the higher one. The activity of acid phosphatase (ACP) and alkaline phosphatase (AKP) increased firstly then decreased. The activity of SOD at 18 and 23 were significantly lower than the control group. The activities of superoxide dismutase at 36 and 40 were significantly higher than that in the control group. The results would provide certain foundation on salinity regulation mechanism of sea cucumber.

Application of Loop-mediated Isothermal Amplification in Detecting Plant Pathogens

QIN Wen-tao, WANG Zhong-yue*, ZHANG Hao*

2013, 15(3):  169-174.  DOI: 10.3969/j.issn.1008-0864.2013.03.24

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Loop-mediated isothermal amplification (LAMP) was a novel method for nucleic acid amplification with the advantages of high specificity, high sensitivity, simple operation, and fitness for detection of varieties environments, and had been widely used in many fields as a molecular biological detection technology. This paper briefly introduced the principle of LAMP, reviewed its application in detecting plant pathogens, discussed the problems occurring in its application and provided relavent solutions according to these problems, and finally outlooked its application prospect.

Establishment and Application of two Rapid ELISA Methods for Detecting Edwardsiella ictaluri

ZHANG Xian-yu1, LI Qiang1*, LI Ming1,2, HUANG Hua1, YE Shi-gen1, LI Hua1

2013, 15(3):  175-182.  DOI: 10.3969/j.issn.1008-0864.2013.03.25

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Two methods including indirect ELISA and competitive ELISA were developed to rapidly detect a pathogen of Edwardsiella ictaluri in the early stage. A monoclonal antibody 5D11 was used as the detection antibody. In indirect ELISA when antigen was coaed at 37℃ overnight, the plate had good adsorption effect on bacteria. The optimal dilution of mAb and AP-goat-anti-mouse IgG were 1∶80 and 1∶1 000, respectively. The lowest concentration of E.ictaluri that could be detected was 5×106 cells/mL. Cross-reactivity test proved that the method had strong specificity. In competitive ELISA, the optimum working combination of coating antigen-mAb was 5×108~1∶80. The proportion of mAb and competitive antigen was 3∶7. A standard curve with R2=0.990 1 was established. This method had strong specificity, and the detection limit was 106 cells/mL. This method can qualitatively and quantitatively detect E.ictaluri within limits, which makes up the deficiency of indirect ELISA.