Table of Content

15 February 2009, Volume 11 Issue 1

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Progress of HarvestPlus China Program

FAN Yun-liu1, WAN Jian-min2, LEI Xin-gen3,4

2009, 11(1):  1-6. 

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HarvestPlus-China(HPC) is a sister or parallel program of HarvestPlus. Its goal is to use plant breeding (biofortification) to reduce or prevent global deficiencies of micronutrients (iron, zinc, and vitamin A) in humans, in particular in developing countries.  The research progress made and the activities  taken by HPC during the past 3 years are summarized, and the problems and prospects of HPC are addressed.

Opportunities Facing Peanut Industry in China and Strategies for its Science and Technology Development

WAN Shu-bo

2009, 11(1):  7-12. 

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This paper analyzes opportunities and challenges facing the  peanut industry development in China; expounds the role played by plant oil safety in promoting peanut production in China, and the trend and characteristics of peanut science and technology development at home and abroad. Combining the scientific and technological development orientation of modern peanut industry, starting from the building of innovative technological system and the key development disciplines for peanut industry, this paper analyzes the trend and respects for science and technology development of peanut industry in the future 20 years. This study will be of important and strategic significance for enhancing the innovative level of China's peanut science and technology and achieving the goal of peanut production with high yield, good quality, high efficiency, ecological balance and food security.

Overseas Research Progress on Energy Crops and Some Focus Issues

YANG Shi-qi, WANG Dao-long,YANG Zheng-li

2009, 11(1):  13-18. 

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Through reviewing and analyzing the development course and present status of energy crops (EC) in certain countries and regions, this paper deems that EC are well developed in America, middle developed in Europe and Australia, and less developed in Asia. On the basis of summarizing the research progress in EC adaptability, ecological effect, cultivation technique, heat energy transformation, research methods and models, this paper puts forward four key issues to develop EC: showing concern on the main body of EC, EC industrialization, environment protection and grain security.

The Application of QDs and its Development Prospect in Plant

PANG Jun-feng1,2| YU Zhuo1, LIANG Zhe2, YIN Cuicui2,3, LIU Hao4,

2009, 11(1):  19-26. 

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Compared with the conventional organic fluorescent dye, quantum dots (QDs) is a new fluorescence material and has the advantages of abundant color, stable photochemical properties, little fluorescence scattering and photobleaching, and low biological toxicity. Consequently, it is widely applied in the fields of  biological tag, human pathology,material science,plant cell detachment and mark,genomics,proteomics,microorganism, biological imaging,biochip and so on. We reviewed the characteristics and research progress of QDs,and its  application in animal,plant and medicine. Meanwhile, we analyzed its essentiality, feasibility and application value in plant research. We also prospected the QDs application in plant, and the detailed research direction.

Research Progress on Hairy Root and its Application

JIANG Yi-na, WU Tian-long

2009, 11(1):  27-32. 

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As a natural genetically engineered strain, Agrobacterium rhizogenes can infect more than 160 plant species, which can lead to special morphological changes of plant and induce a lot of hairy roots production. During the past several decades, people pay more and more attention to research work on hairy roots system, because hairy roots can produce a lot of important secondary metabolites. Mechanization of hairy roots production induced by Agrobacterium rhizogenes was briefly described in this paper. And the recent progress of hairy roots cultivation and application of secondary metabolites were also emphatically introduced.

Biological Function of DNA Methylation

ZHENG Xiao-mei, WU Ning-feng

2009, 11(1):  33-39. 

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DNA methylation, as one kind of modification way of DNA sequences, is an important epigenetic mechanism. It can regulate the expression of many vital genes while the base compositions of these genes do not change, therefore DNA methylation is very important to the growth and development of organism. In this paper, we summarized the rules of DNA methylation into four aspects, as follows: maintaining the stability of genetic information in genomes; regulating the expression of genes; establishing epigenetic patterns and participating in the development of embryos.

Research Progress in the Production of Transgenic Animals Using Spermatogonial Stem Cells

HANG Xin1,2, MIAO Xiangyang1, YIN Xunhe2, MA Yan-fang2,

2009, 11(1):  40-44. 

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Spermatogonial stem cells (SSCs) are kinds of adult stem cells in male mammalia. They are not only the ones capable of selfrenewing and differentiating in testis niche,but also the only ones in normal postnatal body that undergo selfrenewal throughout life and transmit gene to offspring. SSCs have significant value in producing transgenic animals. The origin, formation, type and differentiation of SSCs were reviewed. The application of SSCs in transgenic animals was also summarized.


Review of Water Quality of Miyun Reservoir

WANG Qing-suo, MEI Xu-rong, ZHANG Yan-qing, SUN Dong-bao, YU Yi-lei

2009, 11(1):  45-50. 

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The reservoirs pollution came from soil erosion, waste of animals such as livestock and fish, rubbish and waste water of villages and towns, loss of fertilizer, and nutrient release of the reservoirs deposit. The inorganic pollutants such as total nitrogen and phosphorus in Miyun Reservoir were mainly from Chaohe River basin while organic pollutants such as chemical oxygen demand (COD) and biochemical oxygen demand (BOD) mainly from Baihe River basin. Most of the pollution load came into Miyun Reservoir during the flood season. The concentration of total nitrogen in Miyun Reservoir was higher while that of total phosphorus was lower from 1980 to 2002. Total nitrogen, COD and chlorophyll had an increasing trend year by year. Dissolved oxygen (DO) and transparency (SD) were high, and bacterial, colon bacilli and microcystins were low. Water quality of Miyun Reservoir was in mesotrophic condition before 1990s, and reached mesoeutrophic condition after 1990. The first outbreak of blue green algae in Miyun Reservoir happened during the warm season of 2002.

Thoughts about Agricultural Sci&Tech Park Jointly Constructed by the Academy and Local Government

WANG Xiao-ju1,2, ZHANG Yin-ding2

2009, 11(1):  51-54. 

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This paper probes into the theoretical issues in the fields of cooperation mechanism, development opportunity for jointly constructing agricultural science and technology park by the Academy and local government, its special roles in serving agriculture by sci&tech. It also introduces the practices in developing agricultural high and new technology park of Chinese Academy of Agricultural Sciences. It points out that the joint construction of the park is beneficial to both parties, and in more favor for promoting the construction of modern agriculture and new rural areas.

 |Function Analysis of PhoR/PhoP Twocomponent Regulatory System for Lecithinsolubilizing Ability in Bacillus subtilis Strain NCD-2

LI Jing1,2, GUO Qing-gang2, LI She-zeng2, MA Ping2

2009, 11(1):  55-61. 

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 Bacillus subtilis strain NCD-2 can effectively control cotton verticillium wilt, it also has lecithin (or phosphatidylchoine)solubilizing ability. Plasmid pHV1249 carrying transposon miniTn10 was transferred into strain NCD-2 by electrotransformation, and the transposon insertional mutagenesis was induced at 51℃. miniTn10 insertion mutants were assayed for the lecithinsolubilizing ability, 3 mutants losing lecithinsolubilizing ability were obtained. The flanking sequence of insertion site was cloned from mutant M216 by chromosome walking technique. Sequence analysis showed that the inserted gene was of 98% identity with PhoR of B.subtilis strain 168. Southern blot hybridization proved that strain NCD-2 was inserted by a singlecopy transposon. Genetic complementation was conducted by cloning PhoR into plasmid pHY300PLK and the recombinant plasmid was transferred into mutant M216. The lecithinsolubilizing ability of mutant M216 was restored. The results suggested that the lecithinsolubilizing ability of NCD-2 was related with PhoR gene.

Screening of High Temperature Tolerant Coronatine Producing
Strains by Tn5 Transposon Mutation and Studies
on its Fermentation Conditions

WU Hui-ling, JIAO Rui, ZHANG Jia-chang, LI Zhao-hu

2009, 11(1):  62-67. 

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The plasmid pRL1063a containing Tn5 transposon was transferred into Pseudomonas syingae pv. glycinea PG4180 by conjunction and associators were screened on MG plates with kanamycin and strepmycin. Out of 874 insertion mutants, two mutants which could produce coronatine at high temperature were obtained. They were named as MFB132 and MFB141, respectively. The optimal fermentation conditions for MFB141 were as follows: 2% glucose as carbon source, 0.1% NH4Cl as nitrogen source, the initial pH was 6.8, medium cubage was 50 mL/500mL, and inoculation size was 2%. The fermentation temperature rose from original 18℃ to 28℃ and fermentation time was shortened from 168 hours to 108 hours. The coronatine yield of MFB141 under optimized condition reached to 37.66 mg/L at 28℃, which was 21 times of that of wild type strain (1.8 mg/mL).

Cloning and Expression of Gene Encoding Maltoologosyl Trehalose
Synthase from Corynebacterium glutamicum

ZHANG Wen-de, QIAO Yu, DING Hong-biao

2009, 11(1):  68-72. 

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A 2.4 kb DNA fragment encoding maltooligosyl trehalose synthase was cloned from

Corynebacterium glutamicum by PCR. It was inserted into prokaryotic expression vector

pET-30a and then was introduced into the host Escherichia coli BL21 (DE3) pLysS. A high

yield of the recombinant enzyme at a molecular weight of 90 kDa was obtained by IPTG

induction. The soluble recombinant enzyme accounted for 45.3% of the total cell proteins

in supernatant. This recombinant enzyme was capable of decreasing the DE value of dextrin

and producing a maltooligosyl trehalose mixture.

Effects of Three Different DNA Extraction Methods on the Analysis of
Bacteria Community from Different Micro-ecological Environments
in a Farming Pond by PCR-DGGE

HE Su-xu1, ZHOU Zhi-gang1, YAO Bin1, BAI Dong-qing2

2009, 11(1):  73-79. 

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This study compareed the effects of three different DNA extraction methods (lysozyme digestion, CTAB method and bead mill) on analyzing bacteria from different micro-ecological environments in an farming pond (pond sludge, feed, intestinal content and intestinal wall of grass carp) by PCR-DGGE. The results showed that 16S rDNA V3 fragments were successfully amplified from samples extracted by three DNA extraction methods in all micro-ecological environments; but different DNA extraction methods had remarkable influence on DGGE fingerprints of bacteria under different micro-ecological environments. The cluster analysis of DGGE fingerprints showed that the bacterial community of intestinal content (lysozyme digestion) was almost identical to that of intestinal wall (lysozyme digestion) in grass carp, while the bacterial community of pond sludge (CTAB method) or feed (lysozyme digestion) showed obvious difference to the intestinal bacterial community of grass carp. However, the bacterial community of pond sludge was more similar to intestinal bacterial community compared to that of feed. This study indicated that DNA extraction method should be optimized before analyzing the structure of bacterial community under different micro-ecological environments in an farming pond, and most of the intestinal microbiota in grass carp were derived from the pond sludge in the feed-fed farm pond.

Design and Realization of Nuisanceless Agro-food Information
Publishing System Based on ArcIMS

GAO Tian1,2, LONG Huai-yu2, WANG Jing-kuan1, LEI Qiu-liang2,

2009, 11(1):  80-85. 

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Improving the share of nuisanceless agro-food certification information is an important task at present for Chinas agro-food certification, and taking full advantage of modern information technology to improve the share of agro-food certification information is an important means. According to the status and the characteristic of Chinese nuisanceless agro-food certification, a publishing system based on ArcIMS was designed. This system implemented some functions, including map manipulation, query of agro-food area and agro-food production information, stat. charts creation, short message issuing. With internet as main media, the system can issue the nuisanceless agro-food certification, producing area spatial location, product criterion, and provide a highly effective way to get the nuisanceless agro-food information for the community.

 

Analysis on Cloning and Expression of OsCtBP-A, a C Terminal
Binding Protein (CtBPs) Family Gene of Rice

LI Lei, WU Mao-sen, LI Guang-xu, HE Chen-yang

2009, 11(1):  86-91. 

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A full-length cDNA encoding the C terminal binding protein gene OsCtBP-A, which is down-

regulated during interaction between rice and Xanthomonas oryzae pv. oryzae (Xoo), was

cloned by 5′-RACE. Bioinformatics was employed to elucidate the structure of this gene

and its encoded protein. OsCtBP-A consists of 424 amino acids, with a 2-Hacid_dh_C domain

in N terminal and NLS, PEST motif which are peculiar for plant CtBPs family in C

terminal. Sequence alignment showed OsCtBP-A protein shares a 63% identity with

homologues of Arabidopsis and Ipomoea nil. Furthermore, ABA and SA response elements were

identified within the promoter of OsCtBP-A gene. Real-time quantitative PCR (RTQ-PCR)

showed that OsCtBP-A was induced by salicylic acid (SA), ethane (ETH), methyl jasmonate

(MeJA) and abscisic acid (ABA) treatments with various expression levels respectively.

OsCtBP-A transcript increased drastically with ABA treatment.

Micromorphology and Molecular Marker Identification of
Two Different Types of Lespedeza hedysaroides

YANG Xiu-fang1, SUN Qi-zhong2

2009, 11(1):  92-95. 

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Two different types of L. hedysaroides with yellow-green leaf or dark-green leaf showed

obvious differences in micromorphological features of pod-coat and seed-coat by scanning

electron microscope (SEM) observation. These micromorphological features can be used for

identification of two different types of L. hedysaroides. ISSR-PCR molecular marker was

also used to identify two different types of L. hedysaroides. Among eight primers for

experiments, the primer UBC-880 was able to distinguish the examined two different types

of L. hedysaroides. L. hedysaroides with dark-green leaf has a polymorphic band in

500~600 bp. ISSR-PCR molecular marker can be used to identify two different types of L.

hedysaroides on DNA level.

 

Studies on the Establishment of Genetic Transformation System of
 White Clover（Trifolium repens L.）by Soaking Seeds
 with Agrobacterium tumefaciens

WANG Dan1, WU Yan-min2, LIU Shui3, LV Hui1, LEI Jiang-li1

2009, 11(1):  96-101. 

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The germinating seeds of white clover were used as transformation receptor, and npt-Ⅱ

gene was chosen as a selection gene. External factors that affect the efficiency of

genetic transformation were studied and optimized. A high-frequency transformation system

of white clover had been established by adopting Agrobacterium tumefaciens soaked seeds

transformation technology. The seeds of white clover germinated for 1 hour, then adding

Agrobacterium with concentration of 0.9(OD600) under 5 min+30 s vacuum treatment. After

24 hours infection in oscillating culture and 1 day co-cultivation, the transferred

plants were screened by 50 mg/L kanamycin. This transformation system was so effective,

that positive transformation rate reached 45.5%. The transgenic plants were also examined

by PCR analysis and GUS staining. It was proved that npt-Ⅱ and GUS gene had been

integrated into the genome.

 

Cloning and Analysis of the Terminal Sequence of Barley
Yellow Dwarf Virus GPV

ZHANG Wen-wei, CHENG Zhuo-min

2009, 11(1):  102-107. 

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The authentic 5′ and 3′ terminal sequences of genome of barley yellow dwarf virus

(BYDV) GPV  were obtained by 5′ and 3′ RACE and RT-PCR techniques. The analysis showed

that 5′ terminal sequence in the genome of GPV was 302 nt long, which included the start

codon ATG and 5′ un-translated region (UTR) of 100 nt. Compared with other virus of

Polerovirus, the length of 5′ UTR was variable and un-conservative. While 3′ terminal

sequence in the genome of GPV was 328 nt long, including the terminator codon TGA and 3′

UTR of 93 nt. 3′UTR was 74 nt shorter than that of RPV. The homology of 3′UTR between

GPV and RPV was 40%, but the terminal of 3′UTR was conservative. The homology was 84.34%

between GPV and RPV.

Effects of Soil Type on Seedling Emergence and Growth
of Allium mongolicum Regel

LI Yin-ke, LIU Shi-zeng, HE Fang-yin, ZHANG De-kui

2009, 11(1):  108-112. 

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A pot experiment was carried out to study the effects of 5 soil types including sandy (S), loam (L), sandy loam (SL), upper sandy and lower loam (US+LL), and upper loam and lower sandy (UL+LS) on the seedling emergence and growth of Allium mongolicum Regel. The result showed that the seedling rate of Allium mongolicum Regel had significant positive correlation with soil permeability, seedling rate was the highest in sandy soil (S), due to its good permeability. The seedling growth of Allium mongolicum Regel not only depended on soil permeability, but also related to soil nutrient content. The leaf thickness, root number, fresh weight of shoot and dry weight of shoot were significantly affected by soil types that Allium mongolicum Regel grown in. Growth status of Allium mongolicum Regel in sandy loam (SL) was the best, that in upper sandy and lower loam (US+LL) took the second place.

Effect of Continuous Cotton Cropping on the Microbes and
Enzyme Activities in Soil

SHAN Hong-bin1, LIANG Zhi2, WANG Chun-li1, JIA Hong-tao1, WANG Li1

2009, 11(1):  113-117. 

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Soil microbial qualities and soil enzyme activities were studied in cotton field continuous cropped for 5 years, 10 years, 15 years and 20 years. The results showed that the microorganism amount decreased with the increase of cropping years. Bacterial percentage decreased, while fungi percentage increased from 0.45% to 1.88%. The ratio of B and F decreased. Variation coefficient for actinomycetes, fungi and bacteria were 31.67, 30.8 and 56.12 respectively. Actinomycetes was positive correlation with available K and fungi was negative correlation with available N. Sucrase, urease and phosphatase activities rose in steps with the increase of cropping years. Catalase had no significant differences during the changing process. Sucrase, urease and phosphatase had significant correlation with available K and total N.

Studies on Available Sulfur Content in Soil and its Distribution
Characteristics in Ledong Li Autonomous Country of Hainan Province

ZHAO Xiong, ZHANG Dong-ming| LI Fu-yan| QI Zhi-ping

2009, 11(1):  118-123. 

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787 soil samples of 5 soil types were taken from 11 towns in Ledong county of Hainan province and their available sulfur contents were analyzed. The results showed that the average available sulfur content was 45.24 mg/kg, with the minimum of 3.26 mg/kg and the maximum of 1 296.85 mg/kg. The available sulfur content of most soil samples from all over the county was sufficient, the average available sulfur content was greater than 16 mg/kg, but in some areas of Ledong county the sulfur content was insufficient. The serious lacking of soil available sulfur (content <8 mg/kg) were mostly distributed in Qianjia town and Baoyou town, each accounted for 9.1% and 7.9%. The sulfur content was varied in different soil types, the consequent succession is as followed: coastal sandy soil＞fluvo-aquic soils＞torrid red soils＞paddy soils＞lateritic soils. Sulfur lacking ratio of lateritic soil was 34.46%. The research showed that there was obvious spatial variability of available soil sulfur content in this region.

Initial Studies on Isolation of a Thermophilic Cellulose-degrading
Bacteria and its Enzymology Characteristics

QU Xiao-shuang1,2, DUN Bao-qing1, GUO Ming-ming1,2, SONG Li-li1,3, XU Xiu-hong2,

2009, 11(1):  124-128. 

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WQ-50 was isolated from decayed wood and soil near hot spring in Hainan. It was identified as Bacilluscirculans by preliminary physiological and biochemical characteristics and genetic analysis. When fermented by CMC-Na culture medium, strain WQ-50 strain could grow well under PH 6 and  55℃. Under this condition, when fermented by zymolytic culture medium with alkalized pretreated corn stalk powder as the sole carbon source, enzyme production reached maximum after 5 days, the CMCase activity of WQ-50 is 6.8 IU/mL and FPA activiy of WQ-50 is 3.5 IU/mL.

Studies on Vitamin E in Foxtail Millet Determined by HPLC Method

LI Guo-ying1,2, FAN Zhi-ying3, LIU Fang1,2, ZHANG Ping3, LI Wei-xi1,2,

2009, 11(1):  129-133. 

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Foxtail millet is a food crop rich in vitamin E content. A technology detecting vitamin E content in foxtail millet by applying high performance liquid chromatography（HPLC）was established. The precision, accuracy and reproducibility of this technology were evaluated comprehensively by statistical theory. Afterwards, vitamin E contents of 400 accessions of foxtail millet germplasm were evaluated, and a group of foxtail millets with high vitamin E content was screened.