Scientia Agricultura Sinica

Diversity of Endophytic Bacterial Communities in Seeds of Hybrid Maize (Zea mays L., Nongda108) and Their Parental Lines

LIU Yang, ZUO Shan, ZOU Yuan-Yuan, WANG Jian-Hua, SONG Wei

Scientia Agricultura Sinica. 2011, 44(23): 4763-4771. doi:10.3864/j.issn.0578-1752.2011.23.001

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【Objective】 The objective of this study is to investigate the influence of genotypes of maize on seeds endophytic bacterial communities. 【Method】 Using the 16S rDNA clone library technique, this paper conducted a priliminary study on the community diversity of endophytic bacteria in seeds of reciprocal cross hybrid maize (Zea mays L., Nongda108) and their parental lines. 【Result】 The endophytic bacterial communities in 108A and 108B included 46 and 42 OTUs respectively, and the abundance of the first dominant bacterium Pseudomonas in 108A and 108B was 34.29% and 34.00%. The parental lines Huang C and 178 included 17 and 11 OTUs respectively, the abundance of Enterobacter and Burkholderia as the first dominant bacterium in which respectively was 49.13% and 89.72%. Roseateles, Enterobacter, Pseudomonas and Sphingomonas found in the seeds of 108A were also existed in its female parent Huang C. Pseudomonas, Ralstonia, Paenibacillus and Bacillus were also existed in its male parent 178. Acinetobacter, Pseudomonas, Ralstonia, Paenibacillus and Bacillus found in the seeds of 108B were also existed in its female parent 178. Roseateles, Citrobacter, Pantoea, Pseudomonas, Stenotrophomonas and Escherichia were also existed in its male parent Huang C. 【Conclusion】 In the seeds of reciprocal cross hybrid maize, Nongda 108 and their parental lines which are genetically related, the endophytic bacterial community structure has certain relevance.

Effect of Exogenous L-Phentermine and UV-C on the Accumulation of Rutin Compounds and the Expression of Rutin Biosynthesis Genes in Fagopyrum tartaricum

SUN Chao-Xia, HOU Si-Yu, YANG Wu-De

Scientia Agricultura Sinica. 2011, 44(23): 4772-4780. doi:10.3864/j.issn.0578-1752.2011.23.002

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【Objective】 The objective of this study is to investigate the content of rutin and the expression of the related genes in response to L-phentermine (Phe) and UV-C treatment, and to provide molecular information for high rutin content buckwheat breeding. 【Method】 Rutin biosynthesis-related genes were cloned from the leaves of F. tataricum by 3′RACE. Rutin content in leaves under Phe treatment (1, 2 and 4 mg?L-1, 1 to 5 d) was compared with that of untreated buckwheat by high performance liquid chromatography (HPLC). The gene expression level was analyzed by qRT-PCR. 【Result】 Three partial CDS of rutin biosynthesis-related genes were cloned. They were chalcone synthase (FtCHS), flavonol synthase-like (FtFLS-like) and flavone 3-hydroxylase (FtF3H). Seedlings were cultured in solution containing Phe (4 mg×L-1) for 1 d, rutin content was two times more than the control (12.55 mg×gFW-1), reached 26.15 mg×gFW-1. The higher rutin content (17.36 mg×gFW-1) was found in leaves at 3 h under UV-C treatment. Differential expression patterns of FtCHS, FtFLS-like and FtF3H were observed in different Phe concentration treatments in leaves of F. tartaricum. Seedlings were cultured in solution containing Phe (1 mg×L-1) for 4 d, FtCHS expression level was 4.84 fold higher than the control. The higher expression levels of FtF3H and FtFLS-like were observed after 4 and 2 mg×L-1 Phe treatment for 5 d, they were 1.06 and 3.90 fold higher than the control, respectively. UV-C radiation for 1-3 h, the expression of FtCHS, FtFLS-like and FtF3H was higher than control. In particular, the expression level of FtFLS-like at 1 h under UV-C radiation was 127.71 fold higher than the control. 【Conclusion】 Rutin content could be enhanced to a great extent by Phe treatment and UV-C radiation, and gene expression patterns of rutin biosysthesis-related genes were regulated by UV-C radiation. It can be used to analyze the pathway of rutin biosynthesis. Further study should be focused on the influential factors of rutin metabolic process.

Effects of Different Cultivation Patterns on Grain Yield and Nutrient Absorption and Utilization Efficiency of Japonica Hybrid Rice Changyou 3

XUE Ya-Guang, WANG Kang-Jun, YAN Xiao-Yuan, YIN Bin, LIU Li-Jun, YANG Jian-Chang

Scientia Agricultura Sinica. 2011, 44(23): 4781-4792. doi:10.3864/j.issn.0578-1752.2011.23.003

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【Objective】Rice is one of the most important food crops in China. Continuous realization of high yield has a great significance in ensuring food security and society stability. Meanwhile, low nutrient use efficiency is a serious problem in rice production in China. The objective of this study was to investigate if a cultivation technique could coordinately increase both grain yield and nutrient use efficiency of rice. 【Method】 The japonica hybrid rice cultivar Changyou 3 was used and six cultivation patterns were conducted. No nitrogen application (0N)，local high-yielding cultivation (control), high-yielding and high efficiency cultivation (HHC), super high-yielding cultivation (SHC), super high efficiency cultivation (SHE) and stable yield and high efficiency cultivation (SYHE) were designed. The characteristics of grain yield formation under different cultivation patterns and their nutrient absorption and utilization efficiencies were analyzed. 【Result】 The results showed that the average yields of HHC, SHC,SHE and SYHE were 9.5, 11.5, 10.7 and 9.0 t?hm-2, respectively, and 14.5%, 38.6%, 28.9% and 8.4% higher than the control. Nitrogen recovery efficiency was increased by 39.5%, 93.9%, 86.1%, and 31.0%, and nitrogen agronomic efficiency was increased by 66.5%, 84.4%, 98.2% and 70.1% under HHC, SHC, SHE and SYHE, respectively, when compared with the control. The accumulations of nitrogen (N), phosphorus (P) and potassium (K) from panicle initiation to heading were very significantly and positively correlated with the yields. 【Conclusion】 The results indicated that higher grain yield and higher nutrient use efficiency can be coordinately achieved through integrating and optimizing cultivation techniques in rice production.

Dynamics of Physicochemical Characteristics in Two Waxy Maize Varieties During Grain Filling

LU Da-Lei, YAN Fa-Bao, LU Wei-Ping

Scientia Agricultura Sinica. 2011, 44(23): 4793-4800. doi:10.3864/j.issn.0578-1752.2011.23.004

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【Objective】 The objective of the experiment is to study the dynamics of physicochemical characteristics for waxy maize flour during grain filling. 【Method】 The grain weight, grain volume, water content, starch content, protein content, starch granule size, crystalline type, iodine binding capacity, thermal and pasting characteristics at 10, 20, 30, and 40 days after pollination (DAP) during grain filling were studied using two waxy maize varieties (NQ01 and NQ11) as materials. 【Result】 With the proceeding of grain filling, grain dry weight and starch content gradually increased, water content, protein content and soluble sugar content gradually decreased, while grain fresh weight and volume increased first and fell later. Starch granule size rapidly enlarged from 10 to 20 DAP, while crystalline type and iodine binding capacity presented typical waxy character during grain filling. The results from differential scanning calorimetry (DSC) and rapid viscosity analyzer (RVA) showed that the pasting temperature and transition temperature (onset, peak and conclusion temperature) were gradually decreased, enthalpy of gelatinization was similar from 20 to 40 DAP and lowest at 10 DAP, while the changing tendency of viscosity characteristics and the percentage of retrogradation were different for the 2 varieties. 【Conclusion】Iodine binding capacity, crystalline type and setback presented typical waxy character. The physicochemical characteristics were different during grain filling and the changing of starch granule size and flour quality components content induced the changing of pasting and thermal characteristics, and the dynamics were different between the two varieties.

Coding for Agricultural Product Trace

YU Hua, WU Zhen-Hua

Scientia Agricultura Sinica. 2011, 44(23): 4801-4806. doi:10.3864/j.issn.0578-1752.2011.23.005

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【Objective】An agricultural product trace coding was designed, which can ensure the public to get the enterprise information by the trace coding without professional database.【Method】As each area position code is associated with only one character, the enterprise information is consisted of area position code. Encryption algorithm was applied according to the increasing of code length.【Result】 The trace coding is composed of product code, producing area code, production date code and attestation type code while the producing area code is composed of administrative division code and enterprise name. Producing area code and production date code is converted by decimal-quaternary conversion and dividing section. The validation code is generated by permutation and combination of the identifiers in producing area code and production date code, and attestation type code. The trace coding with thirty-four bits is converted to that with twenty bits.【Conclusion】The design of this twenty trace coding has the characteristics of short code extent and strong encryption. The key information about production enterprise and agricultural products can be traced to locate the production enterprise quickly and accurately without the database and can take emergency measures when the question concerning quality safety of agricultural products happened.

The Complete Genome Sequence of the Gram-Positive Bacterium Bacillus subtils Bs-916

WANG Xiao-Yu, LUO Chu-Ping, CHEN Zhi-Yi, LIU Yong-Feng, LIU You-Zhou, NIE Ya-Feng, YU Jun-Jie, YIN Xiao-Le

Scientia Agricultura Sinica. 2011, 44(23): 4807-4814. doi:10.3864/j.issn.0578-1752.2011.23.006

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【Objective】The objective of this study is to sequence the whole genome of Bacillus subtils Bs-916 and provide more information about molecular biology for future mining and utilizing the potential of this strain. 【Method】 Application of comparative genomics softwares, the whole genome sequence analysis was carried out with Bs168 strains. 【Result】 Bs-916 strains genome is 3 925 958 base pairs comprises 4 056 protein-coding genes. The average GC ratio is 46.4%, the highest compared with other whole-genome sequencing of Bacillus spp.. It contains 152 tandem repeat region, 103 transposons, 37 IS (Insert sequence), 46 tRNA, 39 rRNA. Through comparative genomics analysis, Bs-916 harbors eight giant gene clusters directing synthesis of bioactive peptides and polyketides by modularly organized mega-enzymes named non-ribosomal peptide synthetases, NRPS and polyketide synthases, PKS. Macrolactin, difficidin, and bacillomycin L are absent in Bs168 strains. Bs-916 also contains phytase gene, comAPQX and sfp related with biocontrol mechanism.【Conclusion】Bs-916 strain genome contains many gene cluster encoding a variety of antimicrobial substances, it can be considered a paradigm for an own group of plant-associated gram-positive bacteria with a huge potential for biocontrol and plant growth promotion. The complete genome sequence along with its amenability to genetic manipulation, should facilitate exploitation of the hitherto unappreciated potential of strain Bs-916 to produce secondary metabolites for developing agrobiological engineering preparations.

Behavioral and Lethal Effects of α-terthienyl on the Red Imported Fire Ant (RIFA)

LIU Na, CHENG Dong-Mei, XU Han-Hong, ZHANG Zhi-Xiang

Scientia Agricultura Sinica. 2011, 44(23): 4815-4822. doi:10.3864/j.issn.0578-1752.2011.23.007

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【Objective】 The objective of this study is to define the effects of α-terthienyl on the red imported fire ant (Solenopsis invicta Buren) behavior and lethality.【Method】The behaviors including aggregation, cling ability and walking ability, lethal effect and knock-down activity of α-terthienyl exposed in UVA were researched by potter spray, and the scanning electron microscope was used to probe into the light activity mechanism of α-terthienyl to the red imported fire ant.【Result】When the illumination times were 90, 120 and 180 min, the knock-down rates of the red imported fire ant treated with 100 μg•mL-1 α-terthienyl were 93.08%, 98.29% and 100%, which were higher than those treated without irradiation. After the red imported fire ants were treated with 100 μg•mL-1 α-terthienyl in dark for 90, 120 and 180 min, the knock-down rates of the red imported fire ant were 7.41%, 18.52% and 25.93%, respectively. After they were treated with 100 μg•mL-1 α-terthienyl in irradiation for 30 min, then set into dark for 1, 30, 60, 90, 120, 150 and 180 min, the mortalities of the red imported fire ant were 6.25%, 10.42%, 18.75%, 29.17%, 37.50%, 43.75% and 52.08%, respectively, the walking rates of the red imported fire ant were 25.26%, 18.75%, 10.67%, 6.58%, 2.83%, 2.08% and 0, respectively, the cling rates of the imported fire ant were 20.83%, 12.50%, 6.17%, 4.17%, 2.01%, 1.96% and 0, respectively, and the aggregation rates of the red imported fire ant were 20.78%, 17.19%, 10.94%, 15.63%, 14.06%, 27.08% and 37.50%, respectively. After the red imported fire ants were treated with 100 μg•mL-1 α-terthienyl in irradiation for 30 min and placed in dark for 30 min, the antennal sensilla shape of the red imported fire ant changed significantly. Most of the antennal sensillum presented disordered, lodging and bending phenomen.【Conclusion】 The α-terthienyl possesses the lethal effect and knock-down ability against the red imported fire ant, and can significantlly decrease the aggregation, cling ability and normal walking ability, and also change the shape of the antennal sensilla of the red imported fire ant. The results suggest that the method of using photoactivation composition is expected to become a safe and environment-friendly method for the control of the red imported fire ant.

Dry Matter Accumulation and N, P, K Absorbtion and Utilization in Direct Seeding Winter Oilseed (Brassica napus L.)

LIU Xiao-Wei, LU Jian-Wei, LI Xiao-Kun, BU Rong-Yan, LIU Bo, CI Dan

Scientia Agricultura Sinica. 2011, 44(23): 4823-4832. doi:10.3864/j.issn.0578-1752.2011.23.008

Abstract ( 918 )

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【Objective】An experiment was conducted to study the law of nutrients absorption and accumulation for high yield direct seeded winter oilseed rape (Brassica napus L.).【Method】In order to know N, P and K absorption and utilization of winter oilseed rape which produced 4 500 kg•hm-2 seeds under high yield cultivation patterns, dry matter accumulation and nutrient content and nutrient accumulation in different organs were tested at regular intervals.【Result】Results showed that dry matter accumulation appeared S-type curve and showed as blooming period＞seedling period＞pod period, and roots, stems, green leaves and defoliation reached maximum at 185, 200, 130 and 230 days after seeding and get 2 286, 5 450, 2 306 and 2 162 kg•hm-2 respectively. N content kept stable in seedling period and decreased sharply after flowing period in all organs except seeds. P content increased slowly in seedling period and decreased quickly after flowing in both stems and leaves and decreased constantly in roots, defoliation and shells. K content in both roots and stems changed significantly in seedling period and decreased after then. K content of green leaves decreased in seedling period and kept steadily during other periods. K content in defoliation changed significantly and irregularly. Both N and K accumulation reached maximum in blooming period and decreased little after then. Accumulation of N and K showed as seedling period＞bud period＞blooming period. P accumulation increased constantly in the whole growth period and showed as pod period＞seedling period＞blooming period. The maximum requirement of N, P and K under high yield cultivation were 217.6, 39.9, and 219.8 kg•hm-2, respectively, and the ratio of 1.0﹕0.18﹕1.01.【Conclusion】Dry matter accumulation and nutrient content and nutrient accumulation showed as a falling after rising in each organs except defoliation and reproductive organ. Accumulation of N, P and K in roots and stems and leaves can be recycled in different degrees. Sufficient fertilizer supply before blooming period in winter oilseed rape is the guarantee of high yield.

Applied Research of Combinatorial Algorithm of Clustering，Rough Set and Decision Tree Method in Productivity Evaluation

CHEN Gui-Fen, MA Li, DONG Wei, XIN Min-Gang

Scientia Agricultura Sinica. 2011, 44(23): 4833-4840. doi:10.3864/j.issn.0578-1752.2011.23.009

Abstract ( 968 )

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【Objective】 Fertility evaluation method has a certain subjective and less considers the dependence relation among soil attributes. This paper is aimed to seek a new method of productivity evaluation by data mining method. 【Method】 Based on Nong’an cultivated land survey data, the paper used optimization algorithm of K-means clustering method, Johnson rough set attribute reduction algorithm and C4.5 decision tree algorithm to evaluate the productivity grade. 【Result】 The best learning samples are obtained by using K-means clustering method. Rough sets are used in soil attribute reduction, and 7 soil redundant attributes are removed. The decision tree model has 317 nodes and 159 leaf nodes, extracts 159 rules, model accuracy is 82.08%. The decision tree node number decreased by 41.62% compared with no-clustering and no-reduction approaches. 【Conclusion】 Using the combination algorithm, while the accuracy of the model is ensured, the algorithm time and space complexity are reduced and the mining efficiency is improved.

The Absorption Capacity and Partitioning of 15N to the Major Organs of ‘Fuji’ Apple Trees on M26 Interstock Using 15N-Labelled Urea in Early Summer

HAN Ming-Yu, ZHANG Fang-Fang, ZHANG Li-Xin, LIU Chang-Hong, ZHAO Cai-Ping

Scientia Agricultura Sinica. 2011, 44(23): 4841-4847. doi:10.3864/j.issn.0578-1752.2011.23.010

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【Objective】 The aim of the study is to provide a theoretical basis for N fertilizer rational application for ‘Fuji’ apple under field conditions. 【Method】 The absorption capacity and partitioning of 15N urea of major organs at different phenological stages were conducted using five-year-old field-cultivated ‘Fuji’ apple tree on M26 interstock treated with 15N -urea in early summer. 【Result】 The 15N -urea fertilizer was absorbed and transported by root firstly during the first month. The highest amount of 15N uptaken by root occurred at young fruit stage. The 15N absorption mainly contributed to new organs building with the development of plants in a month. The absorption capacity of 15N urea of new organs was higher in shoot, and then in leaf and fruit. The 15N absorption capacity and partitioning of roots was increased again at harvesting stage. The Ndff% of the major organs was higher in the next year than in the first year in fertilizer. At harvesting stage, the total N content and the amounts of 15N transportation to perennial stem were higher than those to new organs and roots. The 15N distribution ratio in new organs, perennial stem and roots were 23.5%, 40.4%, 36.1% in the first year and 27.7%, 34.9%, 37.4% in the second year, respectively. 【Conclusion】 The N fertilizer application in early summer mainly contented with the requirement to shoots development in the later season and was beneficial for accumulation of nutrition stored in apple trees. The N utilization efficiency of apple tree at fruit harvesting stage was 56.19% in the current year in this study.

Molecular Cloning，Sequence Analysis of Phytoene Synthase Gene from Cerasus humilis (Bge) Sok. and Its Functional Expression in E.coli

ZHANG Jian-Cheng, DU Jun-Jie, LIU He , WANG Peng-Fei, XUE Xiao-Fang, MU Xiao-Peng, CHEN Jun-Qi

Scientia Agricultura Sinica. 2011, 44(23): 4848-4857. doi:10.3864/j.issn.0578-1752.2011.23.011

Abstract ( 636 )

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【Objective】The present study aimed at cloning and analyzing the phytoene synthase (PSY) gene from Cerasus humilis (Bge) Sok. and verifying its catalytic function by heterologous expression in E. coli containing a β-carotene producing plasmid. 【Method】Using the total RNA from the fruit of C. humilis (Bge) Sok. as the template, the cDNA specific fragment of PSY gene was amplified by reverse transcription polymerase chain reaction (RT-PCR), and then the full-length cDNA sequence was obtained through rapid amplification of cDNA ends (RACE) techniques and its sequence was analyzed. The full coding sequence of PSY cDNA was amplified using PCR and further subcloned into vector pET-28a (+). The recombinant plasmid pET-ChPSY was expressed in a prokaryotic expression system after its transformation into BL21 (DE3). The catalytic activity of the fusion protein (6×His-PSY) was investigated in E. coli strain engineered to accumulate β-carotene. 【Result】 Sequence analysis indicated that the full-length cDNA was 1 559 bp, with an open reading frame of 1 194 bp and encoded a protein of 398 amino acids. The cloned cDNA exhibited a homology of the nucleotides and amino acid sequences of over 70% and 65%, respectively, aligned with PSY genes from other plants. The deduced protein has a signal transit peptide consists of 55 amino acid residues in the N-terminal region and two predicted transmembrane domain in the sites of 37-58 and 219-240 amino acids. The analysis of SDS-PAGE demonstrated that the recombinant fusion proteins (6×His-PSY) was produced at a higher level by prokaryotic expression and migrated at a size of about 45.8 kD. The heterogenous expression in E. coli system confirmed that ChPSY could encode a functional phytoene synthase which could enhance β-carotene content of E. coli engineered to produce β-carotene. 【Conclusion】 The PSY gene from Cerasus humilis (Bge) Sok. was successfully cloned and fucntioally expressed in E. coli, which has established a basis for studying the protein characterization and biosynthetic mechanism of carotenoids in fruits of Cerasus humilis (Bge) Sok.

Effects of Phomopsis longanae Chi Infection on Browning and Active Oxygen Metabolism in Pericarp of Harvested Longan Fruits

CHEN Yi-Hui, LIN He-Tong, LIN Yi-Fen, ZHANG Ju-Nian, ZHAO Yun-Feng

Scientia Agricultura Sinica. 2011, 44(23): 4858-4866. doi:10.3864/j.issn.0578-1752.2011.23.012

Abstract ( 765 )

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【Objective】 The effects of Phomopsis longanae Chi infection on browning and active oxygen metabolism in pericarp of harvested longan fruits were investigated in this paper. 【Method】 The harvested longan fruits were inoculated with Phomopsis longanae Chi for 5 min (the concentration was 104 spores per milliliter), while the control fruits were dipped in sterile water. The treated fruits were stored at (28?1)℃ and 90% relative humidity. During fruit storage, fruit disease index, pericarp browning index, malondialdehyde (MDA) content, production rate, activities of active oxygen scavenging enzymes such as superoxide dismutase (SOD), catalase (CAT) and ascorbate peroxidase (APX), contents of endogenous antioxidant substances such as ascorbic acid (AsA) and glutathione (GSH) were determined. 【Result】 As compared to the control fruits there were higher fruit disease index and pericarp browning index of harvested longans infected with Phomopsis longanae Chi. The longan fruits infected with Phomopsis longanae Chi resulted in an increase of production rate and kept higher production rate in pericarp during the whole infection. There were obvious decreases of GSH content and APX activity, whereas, there were sharp increases of activities of SOD and CAT in pericarp of longans during infection from day 0 to day 2. During infection from day 2 to day 5, APX activity of pericarp rapidly increased, while the activities of SOD and CAT declined rapidly. After infection for 5 days, the rapid decreases of activities of SOD and APX in pericarp were observed. The amount of AsA in pericarp significantly decreased while MDA content increased during the whole infection. 【Conclusion】 The results suggest that Phomopsis longanae Chi-induced browning of longan pericarp may be due to a decrease of active-oxygen-scavenging capacity and accumulating large amounts of reactive oxygen species, which may destroy cellular membrane structure, in turn, may cause cellular de-compartmentation, resulted in polyphenol oxidase (PPO) to come into contact with phenolic substrates and oxidation phenolics to form brown polymers.

Effect of Trehalose in Resistance of Wine Yeast to Copper Stress

ZHAO Yu, LI Hai-Lan, DU Jun, ZHAN Ji-Cheng

Scientia Agricultura Sinica. 2011, 44(23): 4867-4873. doi:10.3864/j.issn.0578-1752.2011.23.013

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【Objective】 The accumulation circumstances of trehalose in wine yeast under copper stress and the protection for trehalose to wine yeast were studied in order to provide a theoretical basis for the research on defense mechanism of wine yeast under copper stress. 【Method】 In the present experiment, the modified model synthetic medium (MMSM) was used as yeast culture medium. Two treatment methods, direct-copper stress treatment (1.00 mmol•L-1 Cu2+) and heat shock pre-treatment (37℃, 1 h) with subsequent copper stress treatment were used to treat the S. cerevisia AWRI, S. cerevisia BH8 and S. cerevisia Freddo, respectively. Non-Cu2+ treatment was used as control. The tests determine the survival percentage of wine yeast in different treatment, the accumulation of trehalose and the enzyme activity of trehalose-6-phosphate synthase (TPS1). 【Result】 Compared with the control, 1.00 mmol•L-1 Cu2+ stress treatment increased the contents of trehalose and enhanced the TPS1 enzyme activity. Heat pre-treatment induced the accumulation of trehalose and improved the wine yeast survival percentage significantly more than direct-copper stress treatment. Among the three strains in the same treatment, S. cerevisia Freddo had the most abundant trehalose and highest wine yeast survival percentage. 【Conclusion】 Copper stress treatment could induce the accumulation of trehalose in wine yeast, which consequently serves as a protection for wine yeast under copper stress.

Effects of Diets Supplemented with Nano-CuO and High Level of Zinc Sulfate on Copper and Zinc Contents of Visceral Tissues of Chickens

PAN Na, ZHU Feng-Hua, WANG You-Ling, CHEN Fu, ZHAO Cai-Bing, ZHU Lian-Qin

Scientia Agricultura Sinica. 2011, 44(23): 4874-4881. doi:10.3864/j.issn.0578-1752.2011.23.014

Abstract ( 914 )

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【Objective】 In order to discover the interaction between Zn2+ and nano-CuO in the digestive tract of chickens, an experiment was conducted to determine the effect of high zinc (ZnSO4•7H2O) and normal copper (nano-CuO) diets on copper and zinc contents of visceral tissues in chickens.【Method】One hundred and twenty one-day-old White Leghorn SPF chickens were divided into 4 groups randomly, and five replicates for each group. The basal diet was supplemented with nano-CuO to supply the copper level ats 8 mg•kg-1. The chickens of control group were fed with basal diet. The chickens of trial groups were fed with diets supplemented with ZnSO4•7H2O to supply the zinc level at 250 mg•kg-1, 500 mg•kg-1 and 1 000 mg•kg-1, respectively.【Result】The results showed that when the chickens were fed with diets containing zinc at 250 mg•kg-1, 500 mg•kg-1 and 1 000 mg•kg-1, the zinc and metallotionein contents of liver, proventriculus, duodenum and jejunum were significantly higher than the control group (P＜0.01 or P＜0.05), and the copper contents of duodenum were significantly higher than the control group (P＜0.01). When the diets containing zinc at 1 000 mg•kg-1 were fed to chickens, the copper contents of jejunum were significantly higher than the control group and when the diets containing zinc at 250 mg•mg-1 and 500 mg•mg-1 were fed to chickens (P＜0.01). On 42nd day, when the chickens were fed with diets containing zinc at 500 mg•kg-1 and 1 000 mg•kg-1, the copper contents of liver were significantly higher than the control group and when the chickens were fed with diets containing zinc at 250 mg•mg-1 (P＜0.01). The zinc levels of the diets had a remarkable positive correlation with zinc contents of liver, proventriculus, duodenum and jejunum (P＜0.05), had a remarkable positive correlation with copper contents of duodenum and jejunum (P＜0.05), and had a positive correlation with copper contents of liver (P＞0.05). There was a positive correlation between zinc levels of the diets and the metallothionein contents of duodenum, jejunum and proventriculus (P＞0.05), a significant positive correlation between metallothionein and copper contents of the liver (P＜0.01), and a positive correlation between metallothionein and copper content of duodenum and jejunum (P＞0.05).【Conclusion】It is concluded that the absorption of nano-CuO is not antagonized by high level of zinc of diets supplemented with ZnSO4•7H2O in the intestines of chickens, the nano-Cuo can be absorbed in the form of nanoparticle.

Effects of Dietary Synbiotics on Intestinal Microbial Ecology in Broiler Chickens

YANG Hui-Ling, GAO Yu-Peng, ZHOU Li-Yong, XU Yao-Xing, LI Yan-Li, XU Shao-Chun

Scientia Agricultura Sinica. 2011, 44(23): 4882-4891. doi:10.3864/j.issn.0578-1752.2011.23.015

Abstract ( 915 )

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【Objective】 This study was carried out to investigate the effects of dietary supplementation of synbiotics on intestinal microbial flora ecology in broilers.【Method】Four hundred and fifty 1-d-old Cobb 48 broiler chicks were randomly assigned into 5 dietary treatments, each treatment allocated to 3 replicates of 30 broilers. The dietary treatments were control, basal diets supplemented with antibiotics, basal diets supplemented with probiotics, basal diets supplemented with prebiotics, and basal diets supplemented with synbiotics. On day 21 and 42, four birds from each replicate were randomly selected and killed, cecal contents were aseptically collected, and the total genomic DNA was extracted, then the intestinal bacterial community was analyzed by denaturing gradient gel electrophoresis (DGGE) analysis of universal 16S rDNA after amplication with PCR, and the populations of total bacteria, Lactobacillus, Bifidobacterium and E.coil were detected by real-time PCR. 【Result】 On day 21 and 42, the supplementation of synbiotics increased average daily gain and feed conversion ratio, compared with prebiotics, probiotics or antibiotics alone (P＜0.05), and the number of PCR-denaturing gradient gel electrophoresis bands of synbiotics-fed group was higher than those in the control, antibiotics, probiotics and prebiotics groups. Gene sequences analysis indicated that the specific bands were mainly uncultured bacteria in cecum of broilers. Dietary synbiotics increased the populations of total bacteria, Lactobacillus, Bifidobacterium and E.coil.【Conclusion】The effects of synbiotics on growth performance was consistent with that on intestinal microbial ecology, and the effects of synbiotics was better than probiotics, prebiotics or antibiotics alone.

Effects of Exogenetic Trans-10, Cis-12 CLA on Genes Expression and Protein Synthesis of SREBP-1 Gene in Bovine Mammary Epithelial Cells

WANG Hong-Fang, LIU Hong-Yun, YANG Wei-Ren, LIU Jian-Xin, YANG Zai-Bin

Scientia Agricultura Sinica. 2011, 44(23): 4892-4901. doi:10.3864/j.issn.0578-1752.2011.23.016

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【Objective】The purpose of the study is to explore the effects of exogenetic t10c12 CLA on genes expression and protein synthesis of SREBP-1 in bovine mammary epithelial cells (BMECs). 【Method】 BMECs used in experiment were achieved by tissue culture methods. Lipids in cytoplasm were stained using oil red. The total RNA was extracted from BMECs treated with different gradients of t10c12 CLA. And then the expression levels of various genes were quantified by quantitative reverse transcription PCR (QRT-PCR) analysis. Total proteins extracted with protein extraction kit were used in SDS-PAGE and western blotting. 【Result】Concentration of cytosolic triacylglycerols (TAG) decreased linearly (P=0.0003) with the increasing t10c12 CLA concentration. The addition of t10c12 CLA had no effects on the expression of SREBP-1 gene and other genes related to post-translation process of SREBP-1 and the protein synthesis of precursor SREBP-1(pSREBP-1), maybe, it inhibited the synthesis or promoted the degradation of bio-active SREBP-1(bSREBP-1).【Conclusion】T10c12 CLA didn’t affect the gene expression and protein translation of SREBP-1 and inhibited cytosolic TAG accumulation. According to the results, it was speculated that t10c12 CLA maybe have some effects on activation of pSREB-1 and degradation of bSREBP-1.

Sequence Analysis of mtDNA COⅢ of Tibetan Yaks

ZHAO Shang-Juan, CHEN Zhi-Hua, JI Qiu-Mei, CHAI Zhi-Xin, ZHANG Cheng-Fu, XIN Jin-Wei, ZHONG Jin-Cheng

Scientia Agricultura Sinica. 2011, 44(23): 4902-4910. doi:10.3864/j.issn.0578-1752.2011.23.017

Abstract ( 734 )

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【Objective】The objective of this study was to analyze the genetic diversity and the phylogenetic relationship of different groups of Tibetan Yaks to provide a theoretical basis for protecting genetic diversity of Tibetan Yak and utilization of resources in Tibetan Yak.【Method】The mtDNA sequence of 111 individuals′ COIII from 11 groups of Tibetan Yak was determined by using the Gel Extraction Kit recycling, then the genetic diversity of Tibetan Yak, the affiliation of 11 groups of Tibetan Yak were analyzed. 【Result】 The length of 11 Tibetan Yak groups′ COIII complete sequences were all 781 bp. There were no introns between genes. These genes could encode 260 amino acids. As a start codon, AUG (ATG) had a free base. The average ratios of four nucleotides (T, C, A, G) were 29.2%, 29.4%, 26.1% and 15.2%, respectively. It showed that the base had a bias. It was found that Tibetan Yak’s mtDNA COIII showed 18 haplotypes, the index of haplotype diversity was from 0.378 to 0.844. It showed that Tibetan Yak has rich genetic diversity in mtDNA COIII. These amino acids encoded by COIII, the highest average content is Leu (11.92%), and the lowest one is Lys and Cys (0.77%). The contents of basic amino acid, acidic amino acid, hydrophilic amino acid and hydrophobic amino acid were 11.15%, 4.62%, 29.61% and 54.61%. By phyletic evdution analysis, Tibetan Yak could be divided into three categories, which are Pali (PL) line, Baqing (BQ) line and Sibu (SB) line.【Conclusion】Through cluster analysis，the Tibetan Yak has rich genetic diversity and it could be divided into three major categories. This result supports the theory that Tibetan Yak belongs to a separate genus of bovinae.

Polymorphism Analysis of csd in Five Populations of Chinese Honeybee

LIU Zhi-Yong, WANG Zi-Long, WANG Huan, ZENG Zhi-Jiang

Scientia Agricultura Sinica. 2011, 44(23): 4911-4917. doi:10.3864/j.issn.0578-1752.2011.23.018

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【Objective】 The objective of this study is to analyze the polymorphism of csd in different geographical populations of Chinese honeybee (Apis cerana cerana) (Hymenoptera: Apidae). 【Method】 Worker bees collected from Changbaishan (Jilin), Haikou (Hainan), Nanning (Guangxi), Shennongjia (Hubei) and Jing’an (Jiangxi) were used as materials, and genome DNA was extracted from each sample for PCR amplification of the csd region 3, PCR products were cloned and sequenced, polymorphism of csd in different geographical populations were analyzed by using the obtained sequences. 【Result】 The genomic region 3 of csd in Apis cerana cerana from 5 geographical populations were cloned and 32 csd haplotypes were obtained. The polymorphism of csd in Changbaishan (Jilin) and Jing’an (Jiangxi) populations were significantly higher than those in Nanning, Haikou and Shennongjia populations, while the polymorphism of csd between Changbaishan (Jilin) and Jing’an (Jiangxi) populations, and that between Nanning (Guangxi), Haikou (Hainan) and Shennongjia (Hubei) populations showed no significant difference. Meanwhile, population analysis based on csd haplotypes showed that the nucleotide divergence and genetic distance between Changbaishan (Jilin) and Jing’an (Jiangxi) populations were maximal, while those between Shennongjia (Hubei) and Nanning (Guangxi) populations were minimal. The phylogenetic analysis showed that all the haplotypes from these five geographical populations were mixed on the phylogenetic tree, not from 5 clade according to different geographical origins. 【Conclusion】 csd has a high level of polymorphism in all these populations, moreover, the polymorphism level showed some difference among populations.

Construction of Infectious Clone for Different Genotype Strains of Porcine Circovirus Type 2 and Characterization of the Rescued Viruses in vitro

GUO Long-Jun, LU Yue-Hua, HUANG Li-Ping, WEI Yan-Wu, LIU Chang-Ming

Scientia Agricultura Sinica. 2011, 44(23): 4918-4925. doi:10.3864/j.issn.0578-1752.2011.23.019

Abstract ( 769 )

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【Objective】 Different genotypes (PCV2a, PCV2b and PCV2d) of porcine circovirus type 2 (PCV2) are present in clinical PCV2 infections in China, and it is necessary to elucidate the pathogenic difference among different genotypes of PCV2. 【Method】Representative strains of different genotypes of PCV2 were constructed by infectious molecular clone and biologic characterization of the rescued viruses were identified in vitro. 【Result】 Four infectious clones for different genotypes of PCV2 were constructed and four viruses of different genotypes were rescued after transfection into PK15 cells. The rescued viruses were PCV2 after identified by nucleotide sequence analysis, morphology of the viruses and immunoperoxidase monolayer assay (IPMA). The rescued viruses propagated stably after consecutive incubation for 10 passages, and virus propagation reached up to peak at 72 h post infection (PI), and the virus titers were up to 105.0TCID50/mL. By using neutralizing 1D2 monoclonal antibody (mAb) of PCV2, the antigen capture ELISA showed that only rescued PCV2a/rCL had reactivity with 1D2 mAb, and however, another three rescued strains (PCV2b/rYJ, PCV2b/rJF and PCV2d/rBDH) did not, which indicated the antigenic difference among the rescued viruses of different genotypes.【Conclusion】Conclusion could be draw from this study that PCV2 prevailing has differences in genomic and ORF2 gene length and antigen in swine herds. Four representative clones for different genotypes were constructed and rescued, which will facilitate further studies on the pathogenic difference resulted from different genotypes of PCV2.

Antioxidative Function of Lacbacillus rhamnosus to Caco-2 Cells

CUI Zhi-Wen, HUANG Qin, HUANG Yi, WU Hong-Zhao, WEN Jing, LI Wei-Fen

Scientia Agricultura Sinica. 2011, 44(23): 4926-4932. doi:10.3864/j.issn.0578-1752.2011.23.020

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【Objective】This experiment was conducted to study the antioxidative function of Lactobacillus rhamnosus to Caco-2 cells under oxidative stress. 【Method】 Caco-2 cells were randomly divided into 4 groups, control group and oxidative stress group (100 µmol•L-1 H2O2), treatment groupsⅠandⅡ were added with 1 mL Lactobacillus rhamnosus (about 108 CFU•mL-1) and tertiary butylhydroquinone (TBHQ) (2.75 µg•mL-1 ) under oxidative stress, respectively. Antioxidative activities in culture supernatants and lysates of Caco-2 cells at 12 h and 48 h were measured. 【Result】 Compared to oxidative stress group, the total antioxidation capacity (T-AOC), the glutathione peroxidase (GSH-Px) (P＜0.01), catalase (CAT) (P＜0.01) activities in the cultured supernatants, and the superoxide dismutase (SOD) activity (P＜0.01), and the glutathione (GSH) content (P＜0.05) in the cells lysates at 12 h in treatment groupⅠincreased, and the increases of the antisuperoxide anion radical (O2-) (P＜0.01), GSH-Px (P＜0.01), CAT (P＜0.01), SOD (P＜0.01) activities in the cells culture supernatants and the peroxidase (POD) activity (P＜0.01) and the decreases of malondialdehyde (MDA) content (P＜0.01) in the cells lysates at 48 h were also observed. In treatment groupⅡ, the T-AOC (P＜0.01), CAT activity (P＜0.01) in the cells culture supernatants and the glutathione (GSH) content (P＜0.01) in the cells lysates at 12 h increased significantly, but these values and the MDA content at 48 h decreased significantly (P＜0.01). The anti-superoxide anion radical (O2-), SOD, GSH-Px, POD activities in the cells culture supernatants and the POD activity in the cells lysates at 48 h increased (P＜0.05).【Conclusion】These results indicated that adding Lactobacillus rhamnosus could increase the antioxidative function of Caco-2 cells under oxidative stress under in vitro conditions.

Improving the Independent Innovation Ability in Agricultural Science and Technology

XIN Nai-Quan

Scientia Agricultura Sinica. 2011, 44(23): 4933-4938. doi:10.3864/j.issn.0578-1752.2011.23.021

Abstract ( 654 )

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Under the guidance of scientific development concept, this paper deeply analyzed the current status of agricultural science and technology innovation, for further study and propose the main tasks during the 12th Five-Year-Plan period, and then, discussed the innovation-related theories and problems, i.e., the theories and problems of independent innovation and introduction of advanced technologies, the protection of independent innovation and intellectual properties, the original innovation, integrated innovation, and introduction-utilization-re-innovation, as well as the problems of independent innovation and deepened reform of the systems of science and technologies, independent innovation and fostering the talents and teams, independent innovation and sci-tech input, independent innovation and reform of management systems,etc. with the aims to help start a new ground in agricultural science and technology.

Screening of the Interactive Proteins of Avirulent Proteins AvrPto or AvrPtoB in Susceptible Tomato Cultivar Zhongshusihao by Yeast Two-Hybrid System

ZHANG Yue-Juan, ZHAO Ting-Chang, YANG Yu-Wen

Scientia Agricultura Sinica. 2011, 44(23): 4939-4944. doi:10.3864/j.issn.0578-1752.2011.23.022

Abstract ( 687 )

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【Objective】 The objective of this study is to research the function of AvrPto and AvrPtoB as virulence factor in the susceptible tomato by yeast two-hybrid system. 【Method】 The total RNA of Zhongshusihao was extracted, and dscDNA was amplified by long distance PCR. pGBKT7-AvrPto and pGBKT7-AvrPtoB were constructed. The AvrPto and AvrPtoB interactive proteins were screened by two-hybrid system. 【Result】 In the susceptible tomato cultivar Zhongshusihao cDNA library, 7 proteins interacting with AvrPto were identified, 2 proteins interacting with AvrPtoB were identified. Ribulose bisphosphate carboxylase small chain which can interact with both AvrPto and AvrPtoB showed that AvrPto and AvrPtoB may participate in destroying host photosynthesis. Other proteins such as chloroplastic protoporphyrinogen IX oxidase, chloroplast aldolase which participate in host photosynthesis, and chloroplast elongation factor TufA, transcription elongation factor were also participated in protein synthesis. 【Conclusion】According to the results, it was presumed that AvrPto, AvrPtoB interacting with photosynthesis proteins affected Calvin cycle, the synthesis of chlorophyl related protein. By this way, mesophyll cell of host invation site died.

Sequence Analysis of ITS Gene in Several Amblyseiinae (Acari: Phytoseiidae)

ZOU Zhi-Wen, CHEN Fen, XIA Bin, WU Yu, ZHANG Yu

Scientia Agricultura Sinica. 2011, 44(23): 4945-4951. doi:10.3864/j.issn.0578-1752.2011.23.023

Abstract ( 955 )

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【Objective】The objective of this study is to analyze the phylogenetic relationships among phytoseiid mites in the citrus orchards by comparing their ITS gene fragments, finally improve the classification system of phytoseiid mites. 【Method】The fragment sequences of ITS from 4 phytoseiid mites, Euseius nicholsi, Amblyseius eharai, A. tsugawai and A. orientalis, which collected from citrus orchards in Jiangxi province were analyzed by using softwares.【Result】Length of ITS from the 4 species are from 644-655 bp. The average A+T content is 58.2%. There are 204 variable sites and 32 insertion/deletion in these sequences. Results demonstrated that ITS2 regions are much more conserved than ITS1. Aligned with the ITS sequence of other 5 phytoseiid species of citrus orchard from GenBank, data analysis shows that the relationship between E. nicholsi and E. ovalis is closer than others, their genetic distance is 0.013. Four species of Neoseiulus and 3 species of Amblyseius are clustered into the one branch in the phylogenetic tree, and the nearest genetic distance, which between N. cucumeris and A. orientali, is 0.083.【Conclusion】The results support the taxonomic status of E. nicholsi and E. ovalis, and show sufficient evidence that Neoseiulus and Amblyseius should not be assigned to two genera, which the taxonomic system needs to be further researched.

Effect of Tomato GMPase Overexpression on Tolerance of Potato Plants to Temperature Stress

LI Chao-Han, LI Qing-Zhu, SHI Qing-Hua, BAI Long-Qiang, GUO Xiao-Qing, LI Xia, YU Xian-Chang

Scientia Agricultura Sinica. 2011, 44(23): 4952-4961. doi:10.3864/j.issn.0578-1752.2011.23.024

Abstract ( 703 )

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【Objective】 The aim of the study was to clarify the function of the GDP-D-mannose pyrophosphorylase (GMPase) in the L-galactose pathway of ascorbic acid (AsA) synthesis in plants, and to make sure the relation of GMPase overexpression with ascorbic acid content and temperature-stress tolerance ability. 【Method】 Two transgenic potato strains of overexpressing tomato GMPase were obtained by agrobacterium-mediated transformation. The transgenic plants and wild-type plants were treated in temperature-stress conditions (10/5℃ as low-temperature stress, 35/30℃ as high-temperature stress) for 3 days then in the recover-condition (25/20℃) for 3 days. Real time RT-PCR was used to analyze the expression of GMPase gene. And ascorbic acid content and relative index to temperature stress were also determined. 【Result】Compared with wild-type plants, with the overexpression of GMPase gene in transgenic plants, the GMPase activity and contents of AsA and dehydroascorbate (DHA) in normal and temperature-stress conditions were increased as well as the activities of dehydroascorbate reductase (DHAR), monodehydroascorbate reductase (MDHAR) and ascorbate peroxidase (APX). Meanwhile, the malondialdehyde (MDA) and H2O2 contents and permeability were decreased in the transgenic plants. 【Conclusion】 The overexpression of GMPase may increase potato tolerance to temperature stress by increasing AsA level and lowering reactive oxygen species (ROS).

Analysis on Farmers’ Adoptive Behavior of Soil Testing for Formulated Fertilization: Empirical Evidence from the Xuecheng District of Zaozhuang City in Shandong Province

HAN Hong-Yun, YANG Zeng-Xu

Scientia Agricultura Sinica. 2011, 44(23): 4962-4970. doi:10.3864/j.issn.0578-1752.2011.23.025

Abstract ( 1001 )

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【Objective】 This research was conducted to study the technical results and affecting factors of sequential farmer’s adoption of soil testing for formulated fertilization.【Method】The student’s T test and Bivariate Probit model were employed on the survey data of the sampling area. 【Result】 There was a significant difference in technical results among no-, partial-, and full-adoption of the technology. The area of farmer’s land and technical guidance showed a positive correlation with the farmer’s partial-adoption. The age of farmers had a negative correlation with the farmer’s full-adoption, while the proportion of agricultural income , the area of farmers’ land, concept of fertilization, understanding of the technology, technical guidance, and technical availability card indicated a positive correlation with the farmer’s full-adoption. 【Conclusion】 The farmer’s adoption of soil testing for formulated fertilization had a significant income effect. The most important reasons for no-adoption included equal subdivision of farmer’s land and inconvenience to contact with agricultural technology manager. And the crucial factors in partial-adoption were due to the higher age of farmers, low proportion of agricultural income to personal earning, relatively small arable land, old-fashioned concept of fertilization, incomplete understanding the concept of this novel technology and missing fertilization guidance card.

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