Scientia Agricultura Sinica

The Source of Genes Related to Rice Grain Starch Synthesis Among Cultivated Varieties and Its Contribution to Quality

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Scientia Agricultura Sinica. 2006, 39(05): 865-871 .

Abstract ( 1543 )

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【Objective】The property of starch in rice grain endosperm is a very important determinant for rice quality, and it is essential to understand genetic effect of the genes related to starch synthesis in high-yielding rice varieties for rice quality improvement. 【Method】The physico-chemical properties (e.g. amylose content, gel consistency and RVA profile) were assessed on 53 rice varieties, including some typical indica/japonica landraces and some high-yielding modern varieties. And molecular markers for Sbe1, Sbe3 developed based on sequence diversities between the rice subspecies indica and japonica, together with PCR-AccⅠ marker for Wx gene were used to investigate the genotypes of 53 rice cultivars. 【Results】The result showed that the developed molecular markers for Wx, Sbe1, Sbe3 could distinguish indica or japonica alleles at three loci. Among all the 53 rice cultivars, six genotype combinations were observed at Sbe1, Sbe3 and Wx loci, while WxiSbe1jSbe3i and WxiSbe1jSbe3j genotype were absent. In order to explore the genetic effects of three genes, especially for starch branching enzyme genes, ANOVA and multiple comparison analysis were conducted. It showed that rice cultivars with different genotypes exhibited different phenotypes, including amylose content, gel consistency and some of RVA characteristics, and the significant differences were observed. 【Conclusion】It was concluded that these three genes had randomly recombined during the process of the rice variety development. And the genetic effects of indica and japonica alleles at three gene loci were different, of which, Wx gene plays a major role in determining the starch properties, followed by Sbe1 and Sbe3, and the genetic effects of Sbe1 and Sbe3 in different backgrounds (Wxi, Wxj) are different. The results have provided a clue for rice good quality variety development, and the molecular markers will benefit the rice quality improvement.

The Methodology for Maize Inbred Line with High Yield Selection of and High Combining Ability

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Scientia Agricultura Sinica. 2006, 39(05): 872-878 .

Abstract ( 1796 )

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Based on the inbreeding depression of maize inbred lines, this review addressed the additive effects of gene on heterosis and the correlation between yield and yield components, and analyzed the variation of yields among test crosses of lines in different selfing generations. A methodology that combining the pedigree selection and recurrent selection within the families initiated as early as in S3 was proposed to develop maize inbred lines with high yield and high combining ability.

Genetics, Development and Application of Cytoplasmic Herbicide Resistance in Foxtail Millet

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Scientia Agricultura Sinica. 2006, 39(05): 879-885 .

Abstract ( 1248 )

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【Objective】The effect of cytoplasmic herbicide resistant gene in millet plants was studied. 【Method】The heterozygous populations and isogenic lines with homocaryotic alloplasmic genes were obtained by crossing and reciprocal crossing of cytoplasmic herbicide resistant plants with susceptive plants of foxtail millet. The characters of F1, F2, backcross and composite cross groups and the growth and development of isogenic lines were compared. 【Result】The cytoplasmic herbicide resistant gene slowed the development of seedling, delayed heading and shortened milking stage in foxtail millet plant. Yield capacity and main agronomic characters were all affected by the cytoplasmic herbicide resistant gene in most of the backcross, composite cross and F2 populations. However, there were stronger hybrid vigor in F1. The backcrosses, composite crosses and F2 populations were widely separated and some of them had good characters as that of susceptive groups. 【Conclusion】 The plant characters and development of foxtail millet were negatively affected by the cytoplasmic herbicide resistant gene. The authors proposed a method of using hybrid vigor to obtain high yield and avoid the negative effects of herbicide resistance cytoplasm in plant growth. The expected results could be obtained by selecting individuals in separated populations of fast developed seedlings, well developed roots, and with capacities of early heading and fast milking. Guided by the principal mentioned above, many high yield lines and hybrid crosses of foxtail millet with herbicide resistant cytoplasm were obtained.

Cloning Full-Length cDNA of GbNPR1 Gene from Gossypium barbadense and Its Expression in Transgenic Tobacco

XuJing Wang,,,

Scientia Agricultura Sinica. 2006, 39(05): 886-894 .

Abstract ( 1547 )

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【Objective】The purpose of this study is to find a new gene resource for cotton Verticillium wilt-resistance genetic engineering. 【Method】G. barbadense var. 7124 was used as a starting material to clone the full-length cDNA of NPR1 (none expresser of PR gene)- GbNPR1 which is a key regulator in SA (salicylic acid)-mediated systemic acquired resistance (SAR) by homologous cloning and RACE techniques. 【Result】In the GbNPR1 molecule, there are two "W- boxes" in the upstream of ATG that are necessary for both induction of NPR1 transcription and NPR1-mediated activation of plant defense responses. The deduced amino acid sequence of GbNPR1 has low homology (39%-57%) to the other known NPR1 proteins; however, they have higher homology (79.2%) in the functional domain. GbNPR1 contains the BTB and ankyrin repeat domain that is known to be the molecular basis for NPR1 function in Arabidopsis. Plant expression vector harboring GbNPR1 gene was constructed and transferred into Nicotiana tabacum var. NC89 via Agrobaterium-mediated gene transfer. PCR and Southern-blot analysis indicated that the gene has been integrated into tobacco genome. In vitro leaf-disease challenge test of transgenic plants by inoculation of Alternaria alternata demonstrated that the transgenic plants over expressing GbNPR1 shown enhanced resistance compared with the non-transgenic plants. Genetic analysis of transgenic T1 indicated that inserted gene is one copy. 【Conclusion】All of the results indicated that the obtained gene is the homologous gene of NPR1 gene in G. barbadense.

An Interpretation of the Intellectual Property Aspects of the Golden Rice and Its Implications

Scientia Agricultura Sinica. 2006, 39(05): 895-901 .

Abstract ( 1720 )

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As a world famous transgenic crop developed for a humanitarian target by public institutions, Golden Rice has been incurring, encountering and sustaining various glories, queries, and even blames as well upon its several aspects including the intellectual property rights (IPR). Based on a brief introduction to its technological background, this article presents an interpretation of the IPR issues of the Golden Rice, and manifests that the originally portrayed "giant" IPR obstacle has actually not been so unsurmountable, especially in the target developing countries such as China in which temporarily no relevant patents may exist. In addition, the tragedy of the anticommons in biotechnology has been discussed. Finally, by referring to the actual situation of China, the article gives suggestions for the Chinese scientists and administrators on how to deal with the IPR issues in the biotech research and concludes that China has now been just at a stage of "tragedy of the commons" and not that of the "anticommons".

Structure Characteristics of Stomata in Leaves and Vascular Bundles in Culms of Transgenic Rice Expressing C4 Photosynthesis Enzymes

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Scientia Agricultura Sinica. 2006, 39(05): 902-909 .

Abstract ( 2323 )

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【Objective】The aim of this study is to discover the biological structure basis of transgenic rice expressing C4 photosynthesis enzymes and photosynthesizing highly and efficiently. 【Method】Using four transgenic rice lines over-expressing maize photosynthesis enzymes of phosphoenolpyruvate carboxylase (PEPC), pyruvate, othophosphate dikinase (PPDK), PEPC + PPDK, and rice nicotinamide adenine dinucleotide phosphate-malic enzyme (NADP-ME) as materials, and an untransformed wild type Kitaake as the control, the structures of stomata in leaves and vascular bundles in culms of the seedlings were observed with a scanning electron microscope and mesophyll cells were observed with a transmission electron microscope. 【Result】 Compared with the control, the density and the area of the stomata in leaves of the transgenic rice lines were greater, with the PPDK line greatest among the four lines. The density of stomata in the leaves of transgenic rice with two polymerizing genes (PEPC + PPDK) was increased, but the area of stomata was reduced. The stacking degree of chloroplast thylakoids in mesophyll cells was denser, and some thylakoids were arranged more orderly along the long axis of chloroplast for the transgenic rice lines than those for the WT. The transgenic rice lines exhibited thicker and stronger culms as compared to WT. Except for PPDK, all the transgenic rice lines had larger areas of the tissue structure performing substance-transporting function, such as the outer and inner vascular bundles and their catheter and sieve tubes in culms, as compared with WT. 【Conclusion】 More number and larger area of stomata in leaves and more developed vascular bundles in culms of C4 transgenic rice lines, all these structure characteristics were the basis of its, photosynthesizing highly and efficiently, and closely associated with the greater dry matter accumulation of seedlings.

A Simulation Analysis on Geometrical Parameters of Rice Leaf Blade

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Scientia Agricultura Sinica. 2006, 39(05): 910-915 .

Abstract ( 1317 )

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【Objective】In order to realize more accurate virtual growth of rice, a dynamic model was developed to analyze geometrical parameters of leaf blade based on outputs of rice growth model. 【Method】 Based on the experimental observations, the relationships between the tiller leaf blade length and its synchronously emerging leaf blade length on main stem and between leaf blade length and its width were quantified. Then, coupled with simulation models of leaf age, leaf area and tiller number of rice, a general model for predicting geometrical parameters of rice leaf blade was developed. 【Result】 The ratio of tiller leaf blade length to its synchronously emerging leaf blade length on main stem changed with tiller leaf number in quadratic equation and the relationship between leaf blade length and its width could be expressed in power function. The validation of the model with independent experimental data indicated that the model could simulate the geometrical characteristics of rice leaf blade under different growth conditions. 【Conclusion】 This study has provided a reliable and general method for modeling the geometrical parameters of rice leaf blade and reflected the morpho-physiological patterns of geometrical parameters of crop organs as affected by environmental conditions and variety types.

Relationship Between Leaf Color and Nitrogen Uptake of Rice

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Scientia Agricultura Sinica. 2006, 39(05): 916-921 .

Abstract ( 1425 )

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【Objective】 In order to reveal the relations of leaf color and nitrogen uptake of rice. 【Method】 The dynamic change of upper-most three leaves color and nitrogen uptake rate of rice was studied in solution experiments. 【Result】 There were significant changes of leaves colors and nitrogen uptake during growing period. The three peaks emerged at the days after transplanting, 20 days before heading, and heading and anthesis, low vallies emerged at jointing stage and 10 days before heading. The rate of nitrogen uptake correlated closely with SPAD values of rice leaf after two weeks. 【Conclusion】 The dynamic change of leaf color of rice is mainly caused by nitrogen uptake of root, the changes of leaf color and nitrogen uptake rate are limited by inherent rhythm of its oneself, especially with constant nitrogen concentrations of solution.

The Utility of LAI-2000 Canopy Analyzer Studying the Sunlight Distribution Characteristics in Different Plant Colonies

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Scientia Agricultura Sinica. 2006, 39(05): 922-927 .

Abstract ( 1348 )

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【Objective】 To seek a fast and accurate method for determining the light transmission rate and extinction coefficient.【Method】The LAI, foliar obliquity, canopy openness and light transmission rate in different plant colonies were measured using canopy analyzer LAI-2000 and the solar radiation instruments including the data logger LI-1400, solar radiation sensor LI-190 and LI-191.【Result】 (1) the canopy openness measured by using LAI-2000 had a very close relationship with the sunlight transmission rate of the colonies. The correlation coefficient was 0.9308, the intercept of the regression line was 0.015, which was almost equal to 0, and the slope was 1.0668, which was almost equal to 1. So the canopy openness could be used to estimate the light transmission rate, and the cloudy weather was more suitable. (2) Using the measured LAI and colony light transmission rate, the extinction coefficient of different canopies were calculated accurately. Under the corn canopy, the extinction coefficient changed clearly during the daytime. It had two maximum values and one minimum value. The minimum value happened in the noon.【Conclusion】LAI-2000 canopy analyzer can provide the sky openness of different zenith angles, so it can describe the colony structure in detail; The canopy openness under the canopy is almost equal to the light transmission rate, therefore the measurement of light transmission rate no longer needs the light measurements of over and under the canopy. The LAI of agricultural and forestry canopy measured by LAI-2000 without damaging the colony can be used to calculate the extinction coefficient, and the extinction coefficient calculated in this paper is close to those reported in the references.

Germplasm Development and Mapping of Resistance to Sharp Eyespot (Rhizoctonia cerealis) in Wheat

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Scientia Agricultura Sinica. 2006, 39(05): 928-934 .

Abstract ( 1321 )

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【Objective】Sharp eyespot (Rhizoctonia cerealis) is one of the important diseases in wheat production in China. Due to lack of knowledge on the inheritance of resistance, little progress has been made in wheat breeding for sharp eyespot resistance. 【Method】In this study, the newly evaluated ARz and Niavt 14 were used as sharp eyespot resistant donors, and crossed with widely released cultivars such as Yangmai 158, respectively. At the meantime, ARz/Yangmai 158 genetic population was developed using SSD method with 137 F6 RILs. The 137 RILs and the parents were inoculated with the fungi culture R-46 in F6 using cultured toothpick method and seedbed method, respectively, in 2003.【Result】Novel germplasms, such as 02P12 and 02P315, with multiple resistance and promising agronomic traits were developed. The RIL population showed normal distribution of disease index, with disease index 29.8%-64.4% in cultured toothpick method and 25%-75% in seedbed method, indicating the resistance is a quantitative trait. By using marker regression analysis, 11 SSR markers associated with resistant QTL were found based on the disease index data, explaining 5.0%-13% of phenotypic variance. Among them, Xgwm247, Xgdm67, Xwmc94, and Xgwm437 were detectable in both inoculating methods. QTL mapping of resistance to sharp eyespot was conducted using MapManager QTXb17, 14 linkage groups were established. Two QTLs on chromosome 7D related to the resistance were found by composite interval mapping analysis. One QTL between Xgdm67 and Xbarc172 could been detected, explaining 14.36% and 12.3% of the phenotypic variance, in cultured toothpick and seedbed inoculation conditions respectively. Whereas the another QTL between Xwmc94 and Xwmc273.2 was only been found under the toothpick inoculation, explaining 14.68% of the phenotypic variance. 【Conclusion】 It was deduced that there is a major QTL conferring to resistance to sharp eyespot on 7D in wheat.

The Relationship Between BTH-Induced Resistance to Downy Mildew in Cucumber Seedlings and Content of HRGP and Lignin in Cell Wall

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Scientia Agricultura Sinica. 2006, 39(05): 935-940 .

Abstract ( 1181 )

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【Objective】 The relationship between the activity of phenylalanine ammonia lyase (PAL) or proxidase (POD) and the content of hydroxyproline-rich glycoprotein (HRGP) or lignin and benzothiadiazole (BTH) induced resistance to downy mildew in cucumber seedlings was studied. 【Method】Cucumber seedlings at two-leaf stage were inoculated with spores of Pseudoperonospora cubensis f.sp.cucumerinum before or after treated with BTH. The activities of PAL and POD and the contents of HRGP and Lignin were measured with colorimetric method in the following days. 【Result】BTH treatment not only enhanced the resistance of seedlings to downy mildew, but also increased both enzyme activities and the contents of HRGP and lignin. The accumulation of HRGP and the deposition of lignin synchronized in cucumber leaves after infection of Psedoperonospora cubensis. 【Conclusion】The results suggest that the accumulation of HRGP and the deposition of lignin in cell wall are related to resistant reaction of cucumber to downy mildew, and it is considered a biochemical response of cucumber to downy mildew.

Analysis of Triazophos Residue in Rice by Immunoaffinity Chromatography-High Performance Liquid Chromatography

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Scientia Agricultura Sinica. 2006, 39(05): 941-946 .

Abstract ( 1480 )

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【Objective】The purpose of the study is to enhance the efficiency of triazophos residue monitoring.【Method】The purified anti-triazophos antibody was conjugated to the CDI-activated sepharose CL-4B to synthesize the immunosorbent and prepare the immunoaffinity chromatographic (IAC) column which is specific to triazophos. The conditions of IAC were optimized, pH7.2 phosphate buffer was used as equilibrium and absorbent medium, 60% methanol was used as eluent.【Result】Under the experimental conditions, the dynamic column capacity was up to 1.91 ?g·ml-1 bed volume, the efficiency of enrichment by IAC was nearly 250 times when the initial concentration of triazophos was 2 ng·ml-1 in standard sample solution. The extracts of spiked rice were cleaned up and concentrated by IAC, the triazophos in eluates was determined by high performance liquid chromatography (HPLC), and the average recovery of triazophos was 102.5% with relative standard deviation of 4.44% at the spiked level of 0.1 ?g·g-1. 【Conclusion】The IAC-HPLC technology is developed successfully for the analysis of triazophos residue in rice.

The Connotation and Extension of Agricultural Water Resources Security

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Scientia Agricultura Sinica. 2006, 39(05): 947-951 .

Abstract ( 1365 )

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The objective of this paper is defining the term of agricultural water resources security, its connotation and extension. The definitions of water security, water resources security and water environment security were summarized, and their relations were differentiated and analyzed. Based on these, the elements of the conception of agricultural water resources security were hashed and the conception was defined. Agricultural water resources security should be that water resource provision ensures agriculture against threat, hazard, harm and losing. Moreover, the connotation and extension of agricultural water resources security were ascertained. In detail, the connotation of the definition has natural attributes, socioeconomic attributes and human cultural attributes. The extensions include broad and narrow sense, as well as, food security, agro-environmental security, agro-economic security, rural society security, etc due to agricultural water resources security. The definition will be the frame of reference for developing the researches, limiting the frame of the theory, and founding a appraising system for agricultural water resources security.

Effects of Molybdenum on Antioxidative Enzymes in Winter Wheat Under Low Temperature Stress

Xuecheng Sun,,,,

Scientia Agricultura Sinica. 2006, 39(05): 952-959 .

Abstract ( 1310 )

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【Objective】 Molybdenum is one of the essential micronutrients for plants. The present study was designated to explore the physiological mechanism of cold resistance enhanced by molybdenum in winter wheat.【Method】A pot experiment was conducted to investigate the effects of molybdenum on antioxidative enzymes in winter wheat (Mo efficient cv. 97003 and Mo inefficient cv. 97014) under low temperature stress. 【Result】The activities of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase (APX) increased and the production rates of free radicals decreased in Mo-treated winter wheat under low temperature stress. In Mo-treated winter wheat, with the prolongation of low temperature stress, the activities of the four antioxidative enzymes increased at first 2 or 4 days of low temperature stress, and then decreased or dropped drastically. The tendency in the activities of these antioxidative enzymes indicated that a valid cold acclimation experienced in the Mo-treated winter wheat and that molybdenum helped to maintain the higher level of cold resistance under the following durative low temperature stress. Genotypic differences in response of activities of molybdenum on antioxidative enzymes were found in Mo efficient cv. 97003 and Mo inefficient cv. 97014. The decrease rates of the SOD, CAT, POD and APX activities and the accumulative rate of free radicals for -Mo treatment in Mo inefficient cv. 97014 were higher than those in Mo efficient cv. 97003, which maybe one of the reasons why the cold injury happened frequently in Mo inefficient cultivar for the Mo deficient treatment. 【Conclusion】Molybdenum regulates reactive oxygen species (ROS) metabolism to affect the cold resistance of winter wheat.

Study on Release Characteristics of Different Forms of Nitrogen Nutrients of Slow/Controlled Release Compound Fertilizer

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Scientia Agricultura Sinica. 2006, 39(05): 960-967 .

Abstract ( 1545 )

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【Objective】This paper was to explore the release characteristics of different forms of N nutrients (NH4+-N, NO3--N, urea-N, DON and total N) of uncoated slow/controlled-release compound fertilizer (CRF) and N uptake and use efficiency of rice.【Method】Water dissolution, soil leaching and pot experiment methods were started. 【Result】Results showed that the dynamics of N release of CRF could be quantitatively described by three equations: the first-order kinetics equation (Nt =N0 (1-e-kt)), Elovich equation (Nt＝a+blnt) and parabola equation (Nt＝a＋bt0.5). However, the imitation of the first-order kinetics equation was the best for different forms of N nutrient（r＝0.9569**～0.9999**）. By Nt =N0 (1-e-kt) equation, the release potentials of different forms N (N0 values) in the CRF were in accord with the accumulated nitrogen release amount and in the order of Total N＞DON＞Urea-N＞NH4+-N＞NO3--N in water, and Total N＞NH4+-N＞DON＞Urea-N＞NO3--N in soil. The rate constants of N release (k values and b values) of different forms of N was in the order of Total N＞DON＞NH4+-N＞NO3--N in water. However, the k values was in the order of Urea-N＞DON＞NH4+-N＞Total N＞NO3－-N; and the b values was Total N＞NH4+-N＞DON＞NO3--N＞Urea-N in soil. Compared to the common compound fertilizer (CCF), the N use efficiency, N agronomic efficiency and N physiological efficiency of CRF increased by 11.4%, 8.32 kg·kg-1 and 5.17 kg·kg-1, respectively. The correlation analysis indicated that the ratios (between different forms of N and Total N in CRF) and N uptake by rice were positively significant of very significant. 【Conclusion】The imitationof the first-order kinetics equation (Nt =N0 (1-e－kt)) is the most practical for the release characteristics of different forms of N nutrients. Compared to the only Urea-N of common compound fertilizer, different forms of N nutrients of CRF is in favor of rice.

Studies on Technique of Reducing Methane-Emission in a Rice-Duck Ecological System and the Evaluation of Its Economic Significance

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Scientia Agricultura Sinica. 2006, 39(05): 968-975 .

Abstract ( 1455 )

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【Objective】The rice-duck ecological system is one of the cream practices from the traditional Chinese agriculture. Study on the effect of reducing methane-emission of this practice could provide theoretical and practical basis for further development and utilization of this classical agricultural technique.【Method】The effect of reducing methane-emission and the economic benefits of rice-duck ecological system were studied by adopting a field experiment and economic methodology. 【Result】The daily variation of CH4 emission in late rice paddy field was basically consistent with the daily variation of air temperature. The highest emission occurred at the full tillering stage of late rice with a rate of 24.1 or 32.2 or 40.5 mg·m-2·h-1 in no-tillage area with duck and no-tillage area without duck and conventional-tillage area without duck, respectively. The inhibition of methane-emission was apparently effective in the rice-duck ecological system during the initial tillering stage and the full tillering stage. Compared to the no-tillage area without duck, methane-emission decreased by 2.333 g·m-2. While compared to the conventional-tillage area without duck, methane-emission decreased by 4.723 g·m-2 growth. During the production period of late rice, the amount of methane-emission in no-tillage area with duck was 3.373 g·m-2 less than that of no-tillage area without duck, and 5.59 g·m-2 less than that of conventional-tillage without duck area. The economic significance was analysed. Farmers adopting the rice-duck ecological system obtained 2 166 and 4 207 yuan/ha more income than those who adopted a no-tillage without duck technique or conventional-tillage without duck technique, respectively. In addition to the reduction of the environmental pollution by methane emission, the farmers who adopted the rice-duck ecological system achieved economic benefits of 5 000 yuan/ha, which is 2 206 and 4 274 yuan/ha more than those who adopted a no-tillage without duck technique and a conventional-tillage without duck technique, respectively. 【Conclusion】The rice-duck ecological system not only increased the economic benefits of farmers, but also reduced methane emission in rice paddy field A sustainable agricultural production mode is formed.

Cloning and Expression Analysis of the Self-Incompatibility S9-Haplotype Specific F-box Genes in Cherry (Prunus avium)

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Scientia Agricultura Sinica. 2006, 39(05): 976-983 .

Abstract ( 1401 )

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【Objective】The purpose of this study is to isolate the pollen S-determiant genes involving self-incompatibility in cherry and settle some basic theories for the researches of self-incompatibility mechanism. 【Method】A pair of consensus primers were designed according to the cDNA sequences of 16 S-locus F-box genes published in GenBank. With the RT-PCR and RACE methods, two full length genes coding a kind of polypeptides containing 376 amino acids were cloned in pollen cDNA of cherry cultivar Hongdeng. 【Result】GenBank blast showed that one of the genes coded the same amino acid sequence with the self-incompatibility S3-haplotype-specific SFB (S-locus F-box) gene, SFB3 (AB096857). The other gene coded a novel F-box protein showed no identical sequence in NCBI database and shared 76%-82% amino acid identity with PaSFB1-6. The experimental results showed that the novel gene expressed specifically in pollen and showed the S9-haplotype-specific linkage signal. 【Conclusion】These characters suggested that the novel gene was a new member of pollen S-determinant gene family, PaSFB, in cherry, and was named PaSFB9 (GenBank accession number: DQ422809). The S-genotype of Hongdeng was confirmed as S3S9.

Induction and Genetic Identification of Embryogenic Calli from Hybrids of Shatian Pummelo

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Scientia Agricultura Sinica. 2006, 39(05): 984-989 .

Abstract ( 1092 )

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【Objective】Shatian pummelo (Citrus grandis [L.] Osbeck cv. Shatian) is a peculiar variety in China and it is difficult to regenerate embryogenic callus. Other varieties were used to cross with it in order to gain the callus for further research on cell engineering. 【Method】Shatian pummelo was used as the female parent and crossed with the somatic hybrid NS (Nova Tangelo + Succari Sweet orange, [C. reticulata Blanco × C. paradisi Macf.] cv. Nova + C. sinensis [L.] Osbeck cv. Succari). About 90 days after pollination, the embryos obtained from crosses were cultured on the solid media of MT + ME (malt extraction, 500 mg·L-1) and MT + GA3 (1 mg·L-1). 【Result】The embryogenic callus was induced from the embryoids and plantlets' hypocotyls and can be subcultured. Flow cytometry and SSR analysis verified that the callus was from the triploid hybrid. The callus showed strong capability of embryogenesis and was able to produce a large number of embryoids on MT+ Lactose (50 g·L-1）medium after subcultured for two years.【Conclusion】It is comparatively easy to gain embryogenic callus when Shatian pummelo is crossed with somatic hybrid. The callus is valuable for the conservation and utilization of Shatian pummelo germplasm.

Detection of Staphylococcus aureus in Dairy Products by Polymerase Chain Reaction Assay

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Scientia Agricultura Sinica. 2006, 39(05): 990-996 .

Abstract ( 1236 )

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【Objective】A uniplex polymerase chain reaction(PCR) assay was developed for direct detection of Staphylococcus aureus without enrichment in dairy products. 【Method】A solvent extraction procedure was successfully modified for extraction of Staphylococcus aureus DNA from artifically contaminated whole milk, skim milk and cheese. Primer targeting the thermostable nuclease gene (nuc) was used in the uniplex PCR. A DNA fragment of 279 bp was amplified. PCR product was confirmed by DNA sequencing. In this study, the uniplex PCR, GB 4789.10-94, Perifilm RSA. Count Plate, and Baird-parker + RPF Agar were compared. 【Result】The detection limit of the uniplex PCR is 10 CFU/ml of whole milk, skim milk and 55 CFU/g cheese. The developed methodology allows detection of Staphylococcus aureus in dairy products in less than 6 h, the time of this developed PCR assay is 12-24 h less than the time of general PCR assay with enrichment method, and the coincidence rates of PCR is 94.3%, the sensitivity of PCR is 100%.【Conclusion】The solvent extraction procedure is an effective method for PCR assay of Staphylococcus aureus in milk and milk products.

Research on the Changing Regulation of Ingredients in Cortex Mootan Harvested at Different Stages in Chongqing

Scientia Agricultura Sinica. 2006, 39(05): 997-1003 .

Abstract ( 1431 )

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【Objective】The dynamic changes of multi-ingredients group in Cortex Moutan in Chongqing were studied in order to determine the best growing stage and harvesting time, and to establish a quality control method for Cortex Moutan which is more accordant with herbalist doctor theory. 【Method】 The contents of paeonol in all samples were measured and Contex Moutan samples' chromatographic fingerprints were set up at 273 nm by HPLC. The relative quantity of other ingredients were calculated using paeonol as reference substance in order to understand the dynamic process of the main ingredients in growing circle of Paeonia suffruticosa Andr. by chart analyzing.【Result】(1) The average content of paeonol is 1.25% and the content of paeonol is the highest in samples of Cortex Moutan in Chongqing harvested in Oct. and has grown for 5 years. (2) The contents of other ingredients are the highest in Nov. in the fifth year. Their dynamic changes were positively correlated with paeonol's.【Conclusion】 TCM's multi-ingredients chromatographic fingerprints technology is a modern instrument analysis method which can be used extensively for TCM's quality control. And it can be widely used in producting and circulating field.

Effects of Photoperiod and Melatonin on Nitrogen Partitioning and Cashmere Growth in Inner Mongolia White Cashmere Goats

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Scientia Agricultura Sinica. 2006, 39(05): 1004-1010 .

Abstract ( 1573 )

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【Objective】The effects of photoperiod and melatonin on nitrogen partitioning in Inner Mongolia White Cashmere goats in telogen were investigated. 【Method】Eighteen castrated mature goats, 23-25 kg of liveweight, were divided into three groups randomly, which were treated with different photoperiods, in each of the groups half of the goats were implanted melatonin. Total deposited nitrogen (TN) was tested by general digestive and metabolism method. Body nitrogen deposited (BN) was measured by isotope dilution technique of tritiated water at the beginning and the end of the experiment, fur nitrogen deposited (FN) were calculated by FN= TN- BN. 【Result】Results showed that there was a significant difference in BN and FN partitioning. Hormones relative to nitrogen partitioning and body composition also varied with different treatments. As a result, there was a competitive nitrogen partitioning between body and fur regulated by hormones induced by photoperiod and implanted melatonin. Corresponding to increasing concentration of melatonin in goats blood, there was an additive cashmere in SDPP and implanted groups. Average cashmere production was 338.83±72 g in SDPP and implanted groups, increased by 73.86%. The length of the new grown cashmere was 5.95±0.26-6.32±0.22 cm, the diameter of new cashmere was 14.43±0.28-15.03±0.51 ?m, which was decreased by 0.1-0.71 ?m compared with the previous cashmere. Although cashmere break strength was reduced by 0.94-1.62 cN, it was still in the range of textile standard. Overall, the textile value of new cashmere was up to grade A, except only one group fall in grade B. 【Conclusion】This study has provided evidences that melatonin and photoperiod can be used to improve the cashmere production in practice, the technique can be extended in Inner Mongolia and other areas in north China.

Construction and Immune Responses of the Suicidal DNA Vaccine Co-Expressing GP5 and M of Porcine Reproductive and Respiratory Syndrome Virus

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Scientia Agricultura Sinica. 2006, 39(05): 1011-1017 .

Abstract ( 1465 )

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【Objective】To investigate the immune efficiency of the suicidal DNA vaccines of porcine reproductive and respiratory syndrome virus (PRRSV). 【Method】The ORF5 and ORF6 genes were subcloned into the downstream of two independent subgenomic promoter 26S of a suicidal DNA vaccine expression vector pCSA2 resulting in the plasmid pSFV-56 co-expressing GP5 and M proteins.【Result】The result of Western blot showed that the GP5 and M proteins were expressed and formed disulfided-linked heterodimer. The suicidal DNA vaccine pSFV-56 was injected Balb/c mice and piglets to evaluate the induced immunological responses in vivo. The specific detectable anti-PRRSV neutralizing antibodies were produced in the vaccinated mice at 4 weeks post primary vaccination (PPV) and reached a peak 1:32 at 8 weeks PPV. The specific stronger cell-immunity responses were also observed. In addition, it was observed that the 1:8-1:16 neutralizing antibodies of the vaccinated piglets and the specific cell-immunity responses were produced.【Conclusion】The results of the present study indicated that the suicidal DNA vaccine pSFV-56 has well immunity and the capability inducing higher immune responses in the animals.

Bacteriostatic Action and Mechanism of Sophora flavescens Ait on Escherichia coli 01 C84010

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Scientia Agricultura Sinica. 2006, 39(05): 1018-1024 .

Abstract ( 1422 )

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【Objective】An experiment was carried out to study the bacteriostatic action and mechanism of S.flavescens on E.coli. 01c84010. 【Method】Cylinder-plate and liquid doubling dilution, drafted curves, electron microscope, SDS-PAGE and flow cytometry, were ued in the study.【Results】The result showed that the extracts had obvious bacteriostatic action upon Gram-positive and Gram-negative bacteria and the bacteristatic spectrum was wide. Though restraining the expression of functional protein, the cell cycle was effected and the count of bacteria in phase I increased and that in phase R reduced. Finally, the E.coli 01 C84010 went into decline phase straightly instead of the normal logarithmic phase. Observation under electron microscope revealed that the E.coli 01 C84010, which depressed on center had been constricted into shoe-shaped gold ingots. The cytoplasm was concentrated, the cell was plasmolysed, then the cell wall was damaged, the contents were leaked out and the cell was killed. 【Conclusion】It is suggested that the bacteriostatic mechanism is that the matrine inhibited the synthesis of proteins associated with growth and cleavage of bacteria, then blocked their division and development. Furthermore, the drug combined with the protein, made the cytoplasm pyknosis and killed the bacteria finally.

Effects of Pulmonary Hypertension and Lipid Peroxidation on Pulmonary Vascular Remodeling in Broiler

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Scientia Agricultura Sinica. 2006, 39(05): 1025-1030 .

Abstract ( 1540 )

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【Objective】The changes of pulmonary pressure, lipid peroxidation and microstructure of pulmonary resistance vessel of low or normal temperature treated broilers were investigated to study the mechanism of pulmonary vascular remodeling and its role in the pathogenesis of pulmonary hypertension syndrome (PHS). 【Method】180 Avian commercial broilers were randomly allocated to two groups at 14-day-old age. The group reared under normal ambient temperature served as control group. Another group was exposed to low ambient temperature to induce PHS. The ratio of right ventricular weight to total ventricular weight (RV/TV) and mean pulmonary artery pressure (mPAP) were continuously surveyed. Blood samples were taken to measure the plasma malonilaldehyde (MDA), superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) levels. Lung samples were taken and stained for elastin. The vessel wall area to vessel total area ratio (WA/TA) and mean media thickness in pulmonary arterioles (mMTPA) were measured by using microscopic image analysis software. 【Result】The mPAP, RV/TV ratio and MDA concentration were significantly higher in low temperature treatment group compared with control group. The WA/TA and mMTPA in pulmonary arterioles were significantly increased in low temperature treatment group. Correlation analysis showed that the plasma MDA concentration and mPAP were significantly correlated with WA/TA and mMTPA. 【Conclusion】The results indicated that the promoted lipid peroxidation and pulmonary hypertension in low temperature treated broiler had led to pulmonary vascular remodeling and in turn higher pulmonary artery pressure. These changes seem to be possessed of great importance in the pathologenesis of PHS in broilers induced by low ambient temperature.

Cloning and Bioinformational Analysis of Full-Length cDNA Sequence of Bombyx mori Eukaryotic Translation Initiation Factor 2 Alpha subunit

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Scientia Agricultura Sinica. 2006, 39(05): 1031-1037 .

Abstract ( 1871 )

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【Objective】Clone full cDNA sequence of Bombyx mori eIF2α, analyse cite mutation between susceptible and resistant strain infected by Bombyx mori densovirus-Z (BmDNV-Z). 【Method】Using fluorescent differential display (FDD) technique, a special band named G13-1b650 was obtained in highly susceptible strain HUABA infected by Bombyx mori densovirus-Z (BmDNV-Z). Then electrically extended and further confirmed by using specific primers, a full cDNA sequence of Bombyx mori eIF2α（BmeIF2α）gene was cloned. 【Result】The full length of the obtained cDNA sequence was 1 100 bp (GenBank accession number: DQ073458). Its open reading frame (ORF) includes 999 bp and encodes 332 amino acid residues. The deduced amino acid sequence showed 94.3% identity to eIF2α of Spodoptera frugiperda. It contains one putative conservative phosphorylation site (S（51）R（52）R（52）R（54）R（56）K（60）R（63）), three phosphorylation sites of Protein kinase C, five phosphorylation sites of Tyrosinase, nine phosphorylation sites of Tyrosinase Ⅱ, three phosphorylation sites of protein kinase depended on cAMP or cGMP, two tyrosine sulphation sites and one glycosylation site. 【Conclusion】BmeIF2α in highly susceptible strain contains one tyrosine sulphation site by analyzing sequences from 113-127, H V A E L L H Y E T S E Q S E. While in resistant strains, a C372 to T mutation which leads Serine126 mutaton to Leucine results in the absence of the tyrosine sulphation site in the sequences from 113-127.

Genome Polymorphisms Between Indica and Japonica Revealed by RFLP

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Scientia Agricultura Sinica. 2006, 39(05): 1038-1043 .

Abstract ( 1323 )

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Variation of Indica and Japonica rice were analyzed by using 28 RFLP probers and 6 SSR markers. The result indicated that Indica and Japonica rice can be classified by these markers. The publication of the genome sequences of rice facilitated the development of markers of sub-species and analysis of genome polymorphisms. RZ906 and RM405 markers were analyzed as examples.

An Innovation in New Germplasm of Iron-Enriched Nutritive Health Care Type Super Prematurity Millet

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Scientia Agricultura Sinica. 2006, 39(05): 1044-1048 .

Abstract ( 1174 )

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【Objective】The objective of this study was to solve the difficult problem that adapted crops is single and inefficient in high altitude and cold areas of China.【Method】Using "target character gene bank breeding method", a new millet germplasm Super Prematurity No.2 was developed.【Result】It has some prominent characters: (1) It can normally mature in Bashang of Hebei where the altitude is above 1400 meters. It needs effective accumulated temperature 1650℃ in growing period. (2) The millet is rich in iron and the content of iron is 54.10 mg·kg-1. The average content of iron is 62.0% higher than other varieties in China. (3) The content of crude fat is 6.24%, and the average content of fat is 54.1% higher than other millet varieties. (4) The average content of crude protein of the millet varieties in China is 12.71%, among these varieties, whose crude protein content is higher than 14% is only 5%. The protein content of Super Prematurity No.2 is 14.36%. (5) The synthetic characters are better. 【Conclusion】 It has important significance that super prematurity No.2 has been bred and extended in high altitude and cold areas, thus promoting the development of Super Prematurity millet breeding and the adjusting the structure of crop production in the wide areas with high altitude and low temperature. The practice of super prematurity millet variety breeding indicates that the "targeted-character-gene bank-breeding method" has obvious effects in innovating super prematurity millet and other special idioplasm.

Genetic Diversity of Yunjie (Eruca Mill.) Accessions Revealed by RAPD Marker

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Scientia Agricultura Sinica. 2006, 39(05): 1049-1057 .

Abstract ( 1420 )

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【Objective】The studies of genetic diversity of Yunjie (E. Mill.) accessions and relationships among them are essential for the utilization of them. 【Method】Polymorphism of entire genome of thirty cultivars from Europe, Africa and Asia representing different Yunjie-growing ecotypes was assessed by analyses of RAPD makers. 【Result】Yunjie have abundant polymorphism. Forty-seven out of seventy-four tested random decamer primers were able to identify 486 stable RAPD bands from these Yunjie cultivas. Of them 432 bands, or 89.0% of the total were polymophic. Every group of Yunjie accessions had their characteristic DNA bands. Cluster analysis by unweighted pair group method of arithmetic means（UPGMA）suggested that the 30 Yunjie genotypes could be divided into seven groups, Chinese Yunjie become a group independently and Spain Yunjie belongs to other six different groups. 【Conclusion】The study shows that Spain is richer in Yunjie germplasm diversity than other regions, which is in good agreement with the opinion that west Mediterranean is the origin center and diversity center of Yunjie. The result suggests that China is the second evaluation center of Yunjie.

Cloning Full Length of Ramie Caffeoyl-CoA 3-O-Methyltransferase cDNA and Sequence Analysis

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Scientia Agricultura Sinica. 2006, 39(05): 1058-1063 .

Abstract ( 1731 )

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【Objective】 The full length of ramie(Boehmeria nivea) caffeoyl-CoA 3-O-methyltransferase cDNA was cloned. 【Method】 RACE (Rapid Amplification of cDNA Ends) method was used to analyze the sequence by ClustaW 1.82 software on line.【Result】 The full length of ramie caffeoyl-CoA 3-O-methyltransferase cDNA was cloned for the first time (GenBank accession number: AY 651026). The result of domain analyze showed that it is a member of O-methyltransferases family. Compared ramie mRNA sequence, mRNA CDS sequence and putative amino acid sequence CCoAOMT to other higher plants, phylogenetic tree of CCoAOMT was generated by multiple alignment. The result displayed that mRNA level of ramie can not be clustered into any other plant, but the result showed that ramie and Zea mays (ZMA242980, ZMA242981) have higher homology in mRNA CDS than other higher plants, while the amino acid sequence of ramie and populus (AJ224894, AJ224896, PTU27116) has higher homology than other plants.【Conclusion】Ramie (Boehmeria nivea) caffeoyl-CoA 3-O-methyltransferase cDNA has homology with that of other plants.

Analysis on Fertility Transformation Reaction to Photo-Thermo Treatments of Peiai64S from Different Sources

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Scientia Agricultura Sinica. 2006, 39(05): 1064-1068 .

Abstract ( 1218 )

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【Objective】The characteristics of photo-thermo reaction of Peiai64S lines collected from different sources and different years were analyzed. The change reasons of critical temperature of fertility transformation were discussed.【Method】 The self-fertility and pollen fertility of five Peiai64S lines were analyzed by different photo-thermo treatments in phytotrons.【Result】 Five lines were almost completely sterile under long day light and high temperature (14.5h/28℃), however, under the condition of 14.5h/24℃, except for two lines which kept sterile, the rests were partly fertile, especially the line 4 whose self-fertility rate and pollen fertility was as high as 9.05% and 27.83%, respectively, that was very significant different from the others. The self-fertility of Peiai64S lines collected from the same source but different years were compared and the difference was also found. 【Conclusion】The results indicated that there was a significant difference among Peiai64S lines collected from different regions and different years. The critical temperature of fertility transformation in the line 4 raised and was out of the identification standard of photoperiod and thermoperiod genic sensitive sterile line.

Analysis and Comparison of the Effects of Plastic Film Mulching and Straw Mulching on Soil Fertility

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Scientia Agricultura Sinica. 2006, 39(05): 1069-1075 .

Abstract ( 1538 )

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【Objective】The characteristics of effects of plastic film mulching (PM) and straw mulching (SM) on soil fertility were compared for their selective utilization.【 Method】The effects of PM and SM on soil moisture, temperature, nutrients, and microorganisms were studied in pot and field experiments.【Result】Both of SM and PM had significant moisture-keeping capacity. SM increased soil moisture from the surface to the deep layers, but PM only increased soil moisture in the surface layer and the soil moisture decreased significantly in the deeper layers. PM also increased soil temperature significantly, and the soil temperature in SM treatment was lower than that in CK in the corn growing period, but the soil temperature in PM treatment was lower than that in SM treatment in winter season. SM increased soil organic matter and available N, P, and K to varying degrees and PM didn't, while soil organic matter in the surface layer decreased in PM treatment. SM and PM increased the quantities of bacteria, fungus, and actinomyces in soil, and the increases of soil microorganisms were bigger in SM treatment than in PM treatment. 【Conclusion】Plastic film should be selected when the aim of mulching is to increase temperature. The effect of SM is better than that of PM when the aim of mulching is to keep soil moisture. SM also has the roles of increasing soil nutrients and bio-activity.

Forecast of Heterosis of Introduced Meat Sheep by Genetic Polymorphism of Microsatellite DNA

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Scientia Agricultura Sinica. 2006, 39(05): 1076-1082 .

Abstract ( 1353 )

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【Objective】 Forecast of the heterosis of Small Tail Han Sheep crossed with imported meat sheep by genetic polymorphism of microsatellite DNA was made in different sheep breeds. 【Method】Gene frequency, polymorphism information contents, number of effective alleles, heterozygosity and genetic distances were studied in four imported meat sheep and Small Tail Han Sheep using five microsatellite loci. The crossing effects on Small Tail Han Sheep with four imported meat sheep were tested.【Result】The results indicate that there are genetic polymorphisms at five microsatellite loci in five sheep breeds. Five microsatellite loci can be used for genetic diversity evaluation in sheep breeds. The genetic variability of Dorset is the highest, and Small Tail Han Sheep is the lowest in five sheep breeds. The order of heterosis from large to small in four imported meat sheep by the analysis of genetic relationship is White-Suffolk, Black-Suffolk, Dorset, Texel. It accords with testing results on actual heterosis. 【Conclusion】It is feasible to forecast the heterosis of Small Tail Han Sheep crossed with imported meat sheep by genetic polymorphism of microsatellite DNA and it will have an important value for sheep breeding in future.

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