Scientia Agricultura Sinica

Cloning, Prokaryotic Expression and Quality Effective Analysis of avenin-like Genes from Wheat Cultivars

WEI Hui, DONG Jian, CHEN Qi-Jiao, GAO Xiang, WANG Lei, LI Xiao-Yan, ZHAO Wan-Chun, WU Dan

Scientia Agricultura Sinica. 2012, 45(4): 607-616. doi:10.3864/j.issn.0578-1752.2012.04.001

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【Objective】The present study aimed at cloning, prokaryotic expression and functional analysis of avenin-like genes from common wheat.【Method】The avenin-like genes were cloned by gene specific primers, which designed according to conserved domains of published genes, followed by sequencing and bioinformatic anayzing. Hereafter, JN542444 was cloned into the expression vector, and transformed into the host bacteria Escherichia coli Rosetta-gami B(DE3). After induced by IPTG, the fusion protein was purified by His-Trap affinity chromatography, and then integrated into the control flour to determine the quality effect utilization of 10 g Farinograph. 【Result】 Twenty-two novel avenin-like genes (GenBank accession No. were from JN542443 to JN542464) with 855 bp were isolated by gene-specific primers from 12 wheat cultivars, and all of them encoded 284 amino acid residues. The number and the positions of Cys residues were strictly conservative in 21 genes, which possibly could form 7 intra- and 4 inter-disulfide bonds, except JN542452 with 17 Cys residues. Three pseudogenes, including JN542448, JN542455 and JN542456, were resulted from a single base substitution. An unique type of the avenin-like gene, which was identical to 7 different cultivars in this study and event the other cultivars in GenBank, was named “avenin-like I”. Phylogenetic and evolutionary analysis revealed that avenin-like subunits were closely related to LMW-GS and least homology with ω-gliadin in wheat storage proteins. SDS-PAGE resuslts showed that the gene was successfully expressed in E. coli system. The farinograph data of purified protein revealed that avenin-like I has a significant positive effect on wheat dough quality.【Conclusion】Although lack of polymorphism in the tested cultivars, the Avenin-like I subunit could improve wheat flour processing quality.

Inheritance Analysis of Oleic Acid and Linoleic Acid Content of Arachis hypogaea L.

HUANG Bing-Yan, ZHANG Xin-You, MIAO Li-Juan, LIU Hua, QIN Li, XU Jing, ZHANG Zhong-Xin, TANG Feng-Shou, DONG Wen-Zhao, HAN Suo-Yi, LIU Zhi-Yong

Scientia Agricultura Sinica. 2012, 45(4): 617-624. doi:10.3864/j.issn.0578-1752.2012.04.002

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【Objective】In order to understand the genetic basis of oleic acid content in peanut, the genetic models of oleic acid and linoleic acid was proposed by using F2 populations derived from crosses between normal-oleic and high-oleic genotypes. 【Method】Four cross combinations were made between 4 genotypes with normal oleic acid content and 2 high-oleic genotypes, wt08-0932 and wt08-0934, respectively. The number of major genes controlling oleic acid and linoleic acid contents, their additive and/or dominant effects and heritabilities were estimated using genetic parameters based on genetic models. 【Result】The results indicated that oleic acid and linoleic acid contents were controlled by two major genes with additive-dominant-epistatic effects. The heritabilities of the two major genes were 66%-89% for oleic acid and 70%-85% for linoleic acid, respectively, implying the existence of polygenic effects. Negative dominant effects were observed for both major genes controlling oleic acid content, while positive dominant effects were detected for linoleic acid content. 【Conclusion】The oleic and linoleic acid contents in peanut were controlled by two major genes together with gene interactions and polygene effects. Both of the additive and dominant effects of the first major gene were much higher than that of the second one. The high oleic acid content resulted from the mutation effects of the two major genes and the medium-low and medium-high oleic acid content was the result of the single gene mutation depending on the additive and dominant effects of the individual gene.

Cloning of MaACT3 Promoter from Mulberry (Morus alba) and Agrobacterium-Mediated Transient Expression in Leaves

LI Jun, ZHAO Ai-Chun, WANG Qian-Ling, ZHU Juan-Juan, ZHANG Qiong-Yu, JIN Xiao-Yun, LI Zhen-Gang, LU Cheng, WU Cun-Rong, YU Mao-De

Scientia Agricultura Sinica. 2012, 45(4): 625-632. doi:10.3864/j.issn.0578-1752.2012.04.003

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【Objective】 The objective of this study is to clone the promoter of MaACT3 from mulberry (Morus alba). 【Method】The promoter of MaACT3 was obtained by suppression PCR method and the activity of promoter was valued by agrobacterium-mediated transient expression. 【Result】 The sequence analysis indicated that exon I, intron I and partial exon II were included and the partial exon II was the same as mRNA of MaACT3 reported before. Mature leaves were used for agrobacterium-mediated transient expression. Green fluorescence was measured by fluorescence microscope and mRNA was also detected.【Conclusion】 The promoter of MaACT3 from mulberry expressed promoter activity and could be used in transgenic research of mulberry.

Effects of Sowing Date on Growth Stage and Utilization of Temperature and Illumination of Direct Seeding Rice in Different Ecological Regions

YAO Yi, HUO Zhong-Yang, ZHANG Hong-Cheng, XIA Yan, NI Xiao-Cheng, DAI Qi-Gen, XU Ke, WEI Hai-Yan

Scientia Agricultura Sinica. 2012, 45(4): 633-647. doi:10.3864/j.issn.0578-1752.2012.04.004

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【Objective】 The objective of the study is to elucidate the effect of sowing date on growth stage and utilization of temperature and illumination of direct seeding rice of different varieties in different ecological regions, and to provide a preliminary theoretical basis for the scientific division and distribution of different varieties and its possible sowing date.【Method】Taking late-maturing medium japonica rice, early-maturing late japonica rice and medium-maturing late japonica rice in Taihu rice-growing district in Changshu (Sunan, N31.4°), medium-maturing medium japonica rice, late-maturing medium japonica rice and early-maturing late japonica rice in Lixiahe rice-growing district in Jiangyan (Suzhong, N 32.3°),early-maturing medium japonica rice, medium-maturing medium japonica rice and late-maturing medium japonica rice in Huaibei rice-growing district in Donghai (Subei, N 34.5°) as experimental materials, the effect of sowing date on growth stage and utilization of temperature and illumination of direct seeding rice was comparatively Studied.【Result】The results showed that along with the delayed sowing date, the date of jointing, heading and maturity of all varieties were correspondingly delayed, growth stages significantly reduced that the growth stages reduced by 0.6 days when the sowing date delayed one day. For the same type varieties, the date of jointing, heading and maturity gradually delayed and the growth stages gradually increased with latitude increasing. In the same site, the number of days of growth stages of late japonica rice reduced slightly more than the medium japonica rice with the delayed sowing date. Growth stages reduced with the delayed sowing date because of the vegetative growth period and the vegetative and reproductive growth period reduced which in a range of 2 to 24 days, the reproductive growth period was relatively stable which in a range of -2 to 1 days. The accumulated temperature and illumination hours at growth stages of all varieties reduced significantly or very significantly, which because of the accumulated temperature and illumination hours before heading, especially before jointing, the difference was significant or very significant.In the same site, the accumulated temperature and illumination hours at growth stages of late japonica rice were more than the medium japonica rice. For the same type varieties, with latitude increased, the accumulated temperature reduced, illumination hours reduced except for japonica hybrid rice. The utilization of temperature and illumination of all varieties reduced significantly or very significantly with the delayed sowing date.The number of days from sowing to heading and growth stages increased linearly with the increase of the accumulated temperature and decreased linearly with the increase of illumination hours. The number of days from heading to maturity was different between different treatments with the increase of the accumulated temperature or illumination hours. 【Conclusion】 Sowing date had great effects on growth stage and utilization of temperature and illumination of direct seeding rice. Accordingly, a preliminary division for the direct seeding rice of different variety types was made for providing a reference for the production in the large area.

Study on Spatial Interpolation of Compositional Data Based on Log-Ratio Transformation

LI Chun-Xuan, LUO Yi, BAO An-Ming, ZHANG Yan, YANG Chuan-Jie, CUI Lin-Lin

Scientia Agricultura Sinica. 2012, 45(4): 648-655. doi:10.3864/j.issn.0578-1752.2012.04.005

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【Objective】Spatial interpolation of compositional data needs to meet the four requirements including non-negativity, constant sum, error minimization and unbiased estimation. Soil texture is one of the compositional data. Taking soil texture data of Manas River oasis in Xinjiang as an example, the paper studied the log-ratio transformation approaches on spatial interpolation of compositional data.【Method】First, soil particles content of the soil samples were transformed by using the additive, centered and isometric log-ratio transformation approaches, respectively, then the ordinary kriging method was employed to perform the spatial interpolation. A zero replacement method was introduced to avoid the log-transformation of zero in soil particle composition.【Result】The constant sum of soil particles conent did not change after zero replacement. The kriging based on log-ratio transformation fulfilled the four requirements of compositional data interpolation while the kriging to soil particles separately did not. The kriging based on isometric log-ratio achieved the best results among the three log-ratio approaches, while no obvious differences were found among these approaches.【Conclusion】Zero replacement avoided the log-transformation of zero on the premise of unchanging the requirement of constant sum. The non-negativity, constant sum, error minimization and unbiased estimation of the four requirements of compositional data interpolation could be satisfied by ordinary kriging based on log-ratio transformation.

Effect of Vegetation and Soil on Oviposition of Locusta migratoria tibetensis Chen

LI Qing, LIAO Zhi-Chang, YANG Gang, FENG Chuan-Hong, YANG Qun-Fang, LUO Huai-Hai, JIANG Chun-Xian, WANG Hai-Jian

Scientia Agricultura Sinica. 2012, 45(4): 656-665. doi:10.3864/j.issn.0578-1752.2012.04.006

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【Objective】 The objective of this study is to explore the effect of vegetation and soil physical and chemical properties on fecundity and oviposition distribution of Locusta migratoria tibetensis Chen.【Method】Three-factor quadratic regression general rotary combination experimental design was used to construct oviposition preference model of L. migratoria tibetensis by using the caging simulation method in laboratory and field. 【Result】 L. migratoria tibetensis had oviposition preference on different land soils, and had the highest fecundity in the grassland and barley field (36.03% and 21.20%, respectively). The soil physical and chemical properties had significant effects on oviposition preference. The salt content, water content and pH value had a great effect on oviposition preference. Single factor analysis showed that the fecundity decreased rapidly as the salt content increased in salt content intervals, but the water content and pH value had the optimum value. Double factor analysis showed that the salt content, water content and pH value had interactive effects, and the effect of salt content was the highest. The optimum oviposition soil integrated indictors were found with salt content 0.08%-0.12%, water content 2.91%-3.75% and pH value 7.35-7.65.【Conclusion】L. migratoria tibetensis had a significant difference in fecundity on different vegetations. The salt content, water content and pH value in soil had obvious effects on the oviposition preference of L. migratoria tibetensi.

Effects of Droplet Density and Droplet Size on Control Efficiency of Chlorantraniliprole Against Cnaphalocrocis medinalis (Guenée)

XU De-Jin, GU Zhong-Yan, XU Guang-Chun, XU Xiao-Long, DONG Yu-Xuan

Scientia Agricultura Sinica. 2012, 45(4): 666-674. doi:10.3864/j.issn.0578-1752.2012.04.007

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【Objective】 The objective of this study is to analyze the effect of spray droplet density and droplet size on the control efficiency of chlorantraniliprole against Cnaphalocrocis medinalis under the conditions of different doses, and to provide a scientific basis for efficient use of pesticides in rice field. 【Method】Volumetric application rate was simulated using an automatic drive spraying tower. The surfactant was added to ensure that the chlorantraniliprole solution at different concentrations could provide equivalent wetting and spreading capacity on the rice leaf. The droplet density collected on the water sensitive paper was measured using the image processing method. 【Result】The control efficiency could be improved significantly with the increase of droplet density when the chlorantraniliprole dose was about 2.00 mg•m-2. When the dose was increased to 4.00 mg•m-2 and the droplet density of VMD 200 μm and VMD 75 μm were 26.06 and 66.96 droplets/cm2 respectively, the control efficiency was quite with the higher droplet density treatments. When the droplet density of VMD 200 μm reached 82.09 droplets/cm2, the control efficiency was not reduced with the dose from 4 mg•m-2 to 2 mg•m-2. Likewise, when the droplet density of VMD 75 μm reached 140.06 droplets/cm2, the control efficiency was not reduced with the dose decreased from 4.00 mg•m-2 to 2.50 mg•m-2. Spraying of chlorantraniliprole solution with the same carrier volume and concentration, VMD 75 μm nozzle increased the droplets density and improved the control effects. 【Conclusion】 Droplet density was closely related to the control efficiency of chlorantraniliprole against Cnaphalocrocis medinalis when the effective dose was relatively low. When the droplet density reached a certain quantity, the desired effect could still be obtained when the effective dose was decreased. During low spray volume, the control efficiency could be improved by decreasing droplet size which increased droplet density.

Impact of Spraying Glyphosate on Growth and Yield Component of Glyphosate-Tolerant Soybean at Different Growth Stages

LIU Wen-Juan, LIU Yong, HUANG Xiao-Qin, ZHOU Xi-Quan, SONG Jun, YIN Quan, WANG Dong, TAO Li, ZHANG Fu-Li, CHANG Li-Juan, ZHANG Lei, LEI Shao-Rong

Scientia Agricultura Sinica. 2012, 45(4): 675-684. doi:10.3864/j.issn.0578-1752.2012.04.008

Abstract ( 804 )

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【Objective】 The objective of this study is to research the impact of glyphosate on growth and fecundity of glyphosate-tolerant soybean and to provide statistics and a theoretical basis of time and concentration for weeding control by glyphosate.【Method】A random complete block design was used and the influence of spraying 41% glyphosate isopropylammonium AS at different rates on growth and yield component of transgenic soybean cultivar GTS-40-3-2 at V1-V5 stages was studied in the field. Also, the effect of glyphosate on weeding control was investigated. 【Result】 It was found that glyphosate was effective at 1.23-12.30 kg•ai•hm-2 to control weed. However, the glyphosate more than recommended rate of 1.23-2.46 kg•ai•hm-2 depressed the number of stem nods and compound leaves, and reduced the number of seeds and seeds weight per plant of GTS-40-3-2. There were no obviously negative influence of spraying glyphosate on number of effective pods per plant and 100-seed-weight of GTS-40-3-2, and the main agronomic traits related to yield including plant height, pod height and number of effective embranchments even increased after dealing with glyphosate at lower concentration. There were different effects of spraying glyphosate on growth and fecundity of GTS-40-3-2 at different growth stages. Glyphosate of 4.92-7.38 kg•ai•hm-2 restrained distinctly soybean growth and yield on V1 and V2 stages, but did not injure yield component at V3-V5 stages. The stem nodes and leaves of GTS-40-3-2 were decreased in 10-20 days after treatment (DAT) with glyphosate at V3-V5 stages, then returned to control in 30 d. Glyphosate at 9.84-12.30 kg•ai•hm-2 reduced the number of seeds and seed weight per plant of GTS-40-3-2 at V4 and V5 stages, but did not affect them at V3 stage.【Conclusion】 Spraying glyphosate at 1.23-2.46 kg•ai•hm-2 not only was available for weeding control but also maintained the growth and yield component of GTS-40-3-2, which could be safe to use at V1-V5 growth stages of the glyphosate-tolerant soybean. Application of gyphosate beyond 1.23-2.46 kg•ai•hm-2 depressed the seed number but did not affect the seed mass of GTS-40-3-2. There were differences in growth and yield component depression by glyphosate between growth stages of GTS-40-3-2. Soybean at V1 and V2 stages was more sensitive to glyphosate than V3-V5. It was most safe to spray glyphosate at V3 stage compared with other stages in consideration of soybean growth and fecundity.

Effects of Long-Term Fertilization on Stability of Double Cropping Rice Yield Based on AMMI Model

JI Jian-Hua, LIU Guang-Rong, LI Zu-Zhang, LIU Yi-Ren, HOU Hong-Qian, LIU Xiu-Mei, LI Xu-Hua, LUO Qi-Xiang

Scientia Agricultura Sinica. 2012, 45(4): 685-696. doi:10.3864/j.issn.0578-1752.2012.04.009

Abstract ( 1099 )

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【Objective】 Effects of long-term fertilization on rice yield and its stability of double cropping rice system was studed in this paper.【Method】 Fertilization, environment and their interactions were analyzed by AMMI (additive main effects and multiplicative interaction) model based on a 25-year long-term fertilizer experiment conducted in yellow clayey soil in Nanchang of Jiangxi Province. At this site, 8 treatments were established in 1984. They were in a early rice late rice rotation receiving no fertilizer(CK)，nitrogen fertilizers deficiency (PK), potassium fertilizers deficiency (NP), phosphorus fertilizers deficiency (NK), balanced chemical fertilizers application (NPK), substitution of 70% inorganic N by 30% organic N in balanced chemical fertilizers application (70F+30M), substitution of 50% inorganic N by 50% organic N in balanced chemical fertilizers application (50F+50M)，substitution of 30% inorganic N by 70% organic N in balanced chemical fertilizers application (70F+30M).【Result】Two main results obtained from this analysis showed as follows. Firstly, the sum of squares of fertilization treatments by environment interactions (F×E) was 9.78% of total sum of squares, and achieved a significant difference at 1% level from AMMI model analysis, from which IPCA1, IPCA2, IPCA3 explained 88.46% sum of squares of the interactions. Coefficients of correlations which were significant correlations between the stability parameters and Shukla coefficient of variation or coefficient of variation were 0.97 and 0.83. Secondly, balanced chemical fertilizers application (NPK) treatment could be good for improving the rice yield and its stability of double cropping rice system. Partial substitution of chemical fertilizers by organic manure made further improvement in rice yield and its stability under the equal nutrient amount application condition, compared with NPK treatment. The yields of substitution application of 30%, 50% and 70% of chemical fertilizer by organic fertilizer treatments increased by 6.15%, 3.88%, and 7.75%, respectively, and rice yield stability increased by 25.91%, 59.78%, and 29.31%, respectively. 【Conclusion】 The AMMI model clearly explained the interactive effects of fertilization treatments across environment (F×E). It was a good method for evaluating stability of double cropping rice yield under long-term fertilization. It was concluded that combined application of organic manure and chemical fertilizers was the best fertilization on achieving high and stable yield of double cropping rice in the region. Stability of the treatment combined application of 50% organic manure combined with 50% chemical fertilizers was the best under relatively high yield conditions. Treatments combined application of 30% or 70% organic manure with 70% or 30% chemical fertilizers obtained the highest yield and the best adaptability under the more stable circumstances.

Assessment of Diffusion Gradients in Thin Films Technique for Measurement of the Arsenic Bioavailability in Soils

WANG Jin-Jin, BAI Ling-Yu, ZENG Xi-Bai, SUN Yuan-Yuan

Scientia Agricultura Sinica. 2012, 45(4): 697-705. doi:10.3864/j.issn.0578-1752.2012.04.010

Abstract ( 822 )

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【Objective】This study was conducted to evaluate the feasibility and accuracy of bioavailable arsenic(As) measurement in soils using DGT (Diffusive Gradients in Thin Films Technique) and in further to shed light on the risk evaluation of As-contaminated soils. 【Method】 A pot experiment was used to study the relationship between rape As in rape (Brassica campestris) and the effective concentration(CE) by DGT, soil solution As, NaHCO3-extracted As, NH4Cl-extracted As, and total soil As. Furthermore, the kinetic process of As transporting in soils was simulated using DIFS (DGT Induced Fluxes in Sediments and Soils) model and DGT data.【Result】A significant correlation between As contants in rape (B. campestris) and CE in soils was obtained with the coefficient determination (R2) of 0.990, and the results of DIFS model indicated that the shorter response time (Tc) and a higher desorption rate constants (kb) were both observed in the soils with a greater As bioavailibility. 【Conclusion】 DGT was a reliable and an effective method for pre-concentration of bioavailable As and helped to elucidate the bioavailable process of As in soils on conjugation with the resupply parameters from DIFS model.

Fast Determination of Nitrogen Status in Pear Leaf Using Mid-Infrared Photoacoustic Spectroscopy

ZHAO Hua-Bing, LI Yan-Li, XIE Kai, SONG Xiao-Hui, DONG Cai-Xia, XU Yang-Chun

Scientia Agricultura Sinica. 2012, 45(4): 706-713. doi:10.3864/j.issn.0578-1752.2012.04.011

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【Objective】 The aim was to find a fast and convenient method for prediction of the nitrogen status in pear leaves by technique of spectroscopy.【Method】Pear leaves were collected from 200 dominant pear orchards in four pear experimental stations scattered in three provinces (Shandong, Hebei and Liaoning) around the Bohai Gulf which is affiliated to National Pear Industrial Technology System. Fourier transform mid-infrared photoacoustic spectra (FTIR-PAS) of 200 pear leaf samples were recorded, and Partial Least Squares Regression (PLSR) was applied in the modelings of nitrogen content in pear leaf. 【Result】 The effects of the quantities of Principal Components (PCs) in PLSR and the scope of nitrogen concentrations in samples on the quantitative predicting capacity of PLSR were analyzed. It was found that the established model was of highest accuracy when the quantities of major ingredients in the respective experimental stations, which was referred to Yantai, Tai'an, Changli, Yingkou, and the total samples, as a whole were 6, 6, 6, 4 and 8, respectively. It was also found that when predicting the nitrogen contents in samples by the five established models, Root Mean Square Error of Prediction (RMSEP) was ranging from 1.26 g•kg-1 and 2.18 g•kg-1 and the correlation coefficient of Prediction(Rp) was between 0.644 and 0.806.【Conclusion】The results indicated that it is a new fast and convenient technique for predicting the leaf N status of fruit tree by using the Mid-Infrared Photoacoustic Spectroscopy.

Analysis of Genetic Diversity and Construction of Fingerprint of Vriesea by ISSR

GE Ya-Ying, ZHANG , FEI , SHEN Xiao-Lan, LIU Jian-Xin, WANG Wei-Yong, TIAN Dan-Qing, ZHANG , ZHI

Scientia Agricultura Sinica. 2012, 45(4): 726-733. doi:10.3864/j.issn.0578-1752.2012.04.013

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【Objective】 The purpose of this experiment is to study the genetic diversity and phylogenetic relationship of Vriesea cultivars and establish their DNA fingerprints, which will provide some guidance for crossbreeding and cultivar identification of ornamental bromeliads in Vriesea. 【Method】 Inter-simple sequence repeat（ISSR）was conducted to assess the genetic diversity of 41 Vriesea cultivars and the DNA fingerprints of the cultivars were established.【Result】Among the 41 Vriesea cultivars, 12 ISSR primers produced 132 loci, of which 125 were polymorphisms, and the percentage of polymorphic loci was up to 94.70%. The average value of the effective number of alleles, Nei’s genetic diversity and Shannon’s information index was 1.5311, 0.3101 and 0.4668, respectively. The similarity coefficient between the 41 Vriesea cultivars ranged from 0.3561 to 0.9394. Based on the unweighted pari group method with arithmetic average (UPGMA) and ISSR molecular markers, the 41 Vriesea cultivars were divided into two groups at the similarity coefficient of 0.601. In addition, the DNA fingerprints for the Vriesea cultivars were established by 4 polymorphic ISSR primers.【Conclusion】There existed abundant genetic diversity among 41 Vriesea cultivars and their relationships were significantly correlated with leaf stripe. ISSR molecular markers could be effectively used in genetic diversity and fingerprint analysis for Vriesea cultivars.

Comprehensive Appraisal of the Ornamental Value and Drought and Flooding Resistance of Guoqing Chrysanthemum with Small Inflorescences

DI Li-Li, FANG Wei-Min, CHEN Fa-Di, CHEN Su-Mei, TENG Nian-Jun, GUAN Zhi-Yong, HAN Yong

Scientia Agricultura Sinica. 2012, 45(4): 734-742. doi:10.3864/j.issn.0578-1752.2012.04.014

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【Objective】 The present study was to establish a synthetic system for assessing the ornamental values and drought and flooding resistance of Guoqing chrysanthemum with small inflorescences, and to offer a theoretical support for selection, extension and application. 【Method】 Field and pot-culture experiments were conducted to observe and count 22 indexes of the ornamental values and drought and flooding resistance of 20 Guoqing chrysanthemum varieties with small inflorescences. A synthetic system was established by using analytic hiberarchy process and the K-Means clustering method was applied to differentiate the rank of cultivars.【Result】Twenty-two evaluating factors closely related to ornamental values and drought and flooding resistance were selected and the weight of each affecting factor was calculated. According to the rating criteria, 20 cultivars were endowed with different scores and then multiplied the weight of each factor to obtain the comprehensive score of each cultivars. Finally by K-Means cluster method, using minimum distance method and the clustering center value of synthetical scores of Guoqing chrysanthemum with small inflorescences of various ranks, 20 cultivars were clustered into 3 grades: excellent, good, and general. There were 5, 11 and 4 cultivars in the three grades, and amounted for 25%, 55%, and 20%, respectively. 【Conclusion】 The system constructed by adopting analytic hiberarchy process and cluster method can be used to evaluate and classify small chrysanthemum cultivars. Finally 5 excellent and 11 good cultivars were selected and can be extended in lager areas.

Effects of the Residues of Agricultural Chemicals in Grapes on Winemaking

LI Ji-Ming, SI He-Yun, YU Ying, DUAN Hui, LIANG Dong-Mei, JIANG Wen-Guang, LI Xue-Hui

Scientia Agricultura Sinica. 2012, 45(4): 743-751. doi:10.3864/j.issn.0578-1752.2012.04.015

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【Objective】The residues of agricultural chemicals in grapes and wines from three grapevine varieties in Yantai region were monitored, with the aim to investigate the influence of agricultural chemicals on grape fermentation and wine quality.【Method】The organophosphorous and organochlorine pesticide residues in grape and wine were determined by using gas chromatography-flame photometric detection (GC-FPD) and gas chromatography-electron capture detector (GC-ECD), respectively. 【Result】The striazolone was detected in all grape samples of Cabernet Sauvignon and Chardonnay from four areas in Yantai. Its average content was 71.09 µg•kg-1. Benzene hexachloride was found in Chardonnay from plot 1 and in Cabernet Sauvignon from plot 3, with the concentrations of 2.23 µg•kg-1 and 4.86 µg•kg-1, respectively. Cyhalothrin was found in two grape cultivars from plot 2 and 3, and its concentration ranged from 2.36 µg•kg-1 to 7.51 µg•kg-1 with a mean content of 4.28 µg•kg-1. Among the four agricultural chemicals tested, miconazole nitrate had the strongest inhibitory effect, because the alcoholic fermentation ceased when the amount of this compound exceeded 0.20 g•kg-1. After wine-making, the content of residues of agricultural chemicals in wines were quantified, and benzene hexachloride presented the highest transfer rate from grape to wine (32.14%), whereas chlorothalonil, cyhalothrin and triadimefon showed the values never higher than 0.7%. 【Conclusion】 The residues of agricultural chemicals in wine grapes in Yantai region fulfilled the related requirements. The residues of agricultural chemicals had some effects on fermentation process and generate off-flavors that will negatively affect the sensory of wines. However, the content of residues were reduced evidently during winemaking, and varied significantly in the final products depending on the characteristics of agricultural chemicals.

Hot-Air Treatment Induced Chilling Tolerance of Cold-Stored Chinese Olive Fruits and Its Relation to the Metabolism of Membrane Lipids

KONG Xiang-Jia, LIN He-Tong, ZHENG Jun-Feng, LIN Yi-Fen, CHEN Yi-Hui

Scientia Agricultura Sinica. 2012, 45(4): 752-760. doi:10.3864/j.issn.0578-1752.2012.04.016

Abstract ( 1057 )

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【Objective】 The effects of hot-air treatment on chilling tolerance of cold-stored Chinese olive fruits in relation to the changes of lipoxygenas (LOX) activity and fatty acid constituents of membrane lipids were investigated. 【Method】 The harvested ‘Tanxiang’ Chinese olive [Canarium album (Lour.) Raeusch cv. Tanxiang] fruits were pre-treated with hot-air at 38℃ for 30 minutes and then packed into sealed polyethylene bags (0.015 mm thickness) and stored at (2?1)℃. Chilling injury index and cellular membrane permeability of fruit, LOX activity and fatty acid constituents of membrane lipids in pericarp of Chinese olive fruits during cold storage were determined.【Result】As compared to the control fruit, there were lower fruit chilling injury index, cellular membrane permeability and pericarp LOX activity in cold-stored Chinese olive fruits pre-treated with hot-air at 38℃ for 30 minutes. The contents of unsaturated fatty acids in pericarp, such as palmitoleic acid (C16:1), linoleic acid (C18:2) and linolenic acid (C18:3) increased, while the contents of saturated fatty acids in pericarp, such as myristic acid (C14:0), palmitic acid (C16:0) and stearic acid (C18:0) decreased, and index of unsaturated fatty acids (IUFA) and unsaturated degree of fatty acids in pericarp increased. 【Conclusion】It is suggested that the enhancement of chilling tolerance and the alleviation of chilling injury in cold-stored Chinese olive fruits by hot-air treatment may be due to the decrease of LOX activity in pericarp, which may reduce the degradation of unsaturated fatty acids of membrane lipids and maintain higher unsaturated degree of fatty acids.

Effect of Dietary Urea Supplementation Level on Growth Performance and Blood Biochemical Indices in Growing-Finishing Cattle

JIANG Lan, MENG Qing-Xiang, REN Li-Ping, HUO Yun-Long, WANG Li-Wen, DING Jian, ZHAO Jin-Wei

Scientia Agricultura Sinica. 2012, 45(4): 761-767. doi:10.3864/j.issn.0578-1752.2012.04.017

Abstract ( 799 )

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【Objective】A feeding trial was designed to investigate the effect of dietary urea levels on growth performance and blood biochemical indexes of growth-finishing cattle for providing a theoretical basis for the optimum level of urea supplementation in beef cattle diet.【Method】 Sixty growing-finishing Limousin×Fuzhou crossbred bulls were randomly selected and divided into six groups with 10 bulls in each treatment group. The treatment diets were six supplemental urea levels (0, 0.4, 0.8, 1.2, 1.6 and 2.0%) in the complete diet and contained similar CP and ME concentrations (14%CP, 11.3 MJ•kg-1 on DM basis). The trail was lasted for a total of 14 wks including 1 wk for adaptation and 13 wks for data collection. 【Result】 As dietary urea level increased, daily DM intake was not different significantly (P＞0.10), but ADG was linearly decreased (L, P＜0.05), and feed conversion efficiency (DMI/ADG) was linearly increased (L; P＜0.05). There was no significant response in ADG and feed conversion efficiency (DMI/ADG) within 0.8% of dietary urea level. Increasing dietary urea-N level resulted in remarkable increases in plasma ammonia (L, P＜0.001), total proteins (Q, P＜0.001) and urea-N (Q; P＜0.05) with break points of total plasma proteins and plasma urea-N appearing at 1.50% and 0.64% of dietary urea supplementation level, respectively. When dietary urea level increased, blood alanine aminotransferase (ALT) activity was unchanged significantly (P＞0.1), but aspartate aminotransferase (AST) activity linearly increased (L, P＜0.001). Increasing dietary urea-N level quadratically (Q, P＜0.05) affected the concentrations of blood biochemical indices in relation to stress reaction, such as the activity of serum creatine kinase, concentrations of norepinephrine (NE), corticosteroid (COR) corticosterone and aldosterone (ALD) except epinephrine (E), with altered trends obtained between 0.8%-1.2% of dietary urea supplementation level. 【Conclusion】 Considering the result of the growth performance and blood biochemical indices, urea inclusion level of lower than 0.8% in the growing-finishing diet DM was be recommended as an optimal and safe level, which is equivalent to 16% of urea-N of total dietary N.

Genetic Variation of ESR Gene in Different Generations of Sujiang Pigs and the Impact on Reproductive Traits of Pigs

WANG Xiao-Yan, HE Qing-Ling, JING Rong-Bin, SONG Cheng-Yi

Scientia Agricultura Sinica. 2012, 45(4): 768-773. doi:10.3864/j.issn.0578-1752.2012.04.018

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【Objective】This study was aimed to analyze PvuⅡ polymorphism of ESR gene in different generations of Sujiang pigs and the correlation between the polymorphism and the reproductive traits of Sujiang pigs. 【Method】 PCR-RFLP was used to diagnose polymorphism of ESR gene in 906 Sujiang pigs of 2-6 generations. Least square method and one-way anova were performed to evaluate the association of this polymorphism site with reproductive traits of pigs. 【Result】 The frequency of allele B of ESR gene in Sujiang pigs of 2-6 generations showed an increasing trend. Pigs with BB genotypes appeared in the third generations and then presented an upward trend in the frequency. There was no significant relationship between the polymorphism of ESR gene and litter size of sows mated to different genotypes of boars. But they had significant impacts on litter size that different genotypes of boars mated with different genotypes of sows (P＜0.05). 【Conclusion】 The selection of boars for reproductive traits of Sujiang pig must be taken into consideration.

Analysis of Sequence Characters of ADSL Gene and Correlation between Gene Expression and IMP Content in Duck

XU Shan-Jin, YU De-Bing, WANG Feng, HE Zong-Liang, ZHANG Jian-Feng, YU Hai-Long, DU Wen-Xing

Scientia Agricultura Sinica. 2012, 45(4): 774-785. doi:10.3864/j.issn.0578-1752.2012.04.019

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【Objective】The CDS (coding sequences) of duck ADSL gene was cloned. The study was aimed to analyze gene sequences, protein structure, relative expression levels of ADSL gene in muscle and the correlation between gene expression and inosine monophosphate (IMP) content in ducks. This research revealed the ADSL gene structure of excellent native ducks of China, established a foundation for further research of its biological function and relationship with flavor substances. 【Method】 Reverse transcriptase PCR was employed to clone the cDNA of ADSL gene. The coding region sequences, protein structure and phylogenetic relationships were analyzed by bioinformatics software. real-time-PCR was employed to examine the relative expression levels of ADSL gene in muscle of ducks. IMP content in breast and leg muscle was detected by HPLC. 【Result】 The CDS of ADSL gene in ducks had 1380 bp in length, encoded 459 amino acid residues and showed 94.77% sequence identity with chicken. The physical and chemical properties of duck ADSL indicated that ADSL is a basic protein. And the protein has more strongly signal peptide and transmembrane segments in cells. The secondary structure of duck ADSL was mainly made by α helixes. Phylogenetic analysis showed that duck and chicken were in the same clustering, and then clustered with human, mice and other mammals, both of which had a distant relationship with fish and amphibian. The expression levels of ADSL gene among different varieties were extremely different (P＜0.01), that between female and male were extremely different (P＜0.01), and that among different locations were different (P＜0.05) except those in cherry valley ducks (P＞0.05). IMP content among different varieties were extremely different (P＜0.01), and among different sexes and locations were very different (P＜0.05). 【Conclusion】 The CDS of ADSL gene in ducks showed a high sequence identity with chicken. The expression levels of ADSL gene and the IMP content of relative locations were extremely different (P＜0.01) or different (P＜0.05) among different varieties, sexes and locations, respectively. ADSL gene relative expression and IMP content were significantly correlated.

Differential Proteome Analysis of the MARC-145 Cell Infected with Highly Pathogenic PRRSV

ZHOU Lun-Jiang, WANG Long-Bai, FANG Qin-Mei, WU Xue-Min, CHE Yong-Liang, CHEN Ru-Jing, WEI Hong, ZHUANG Xiang-Sheng

Scientia Agricultura Sinica. 2012, 45(4): 786-793. doi:10.3864/j.issn.0578-1752.2012.04.020

Abstract ( 808 )

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【Objective】 The objective of this study is to analyze the protein expression changes of the MARC-145 which was infected with highly pathogenic PRRSV. Proteomic techniques were used to establish the protein expression profiles. The study provides the global information of proteomic alteration of MARC-145 cells in the presence of intact PRRSV and the clues for further understanding of the mechanism of interaction. 【Method】 The differentially expressed proteins between the MARC-145 cells that were infected with highly pathogenic PRRSV and the normal MARC-145 cells were profiled and identified by two dimensional electrophoresis and Western Blotting. Differential proteinaceous information was identified by mass chromatographic analysis and retrieval. 【Result】The results showed that there were seven differentially expressed proteins. Five proteins were identified successfully which were heat-shock 70-kD protein, thioredoxin, S100 calcium-binding protein A6, glyceraldehyde-3-phosphate dehydrogenase, and beta-actin, respectively. Among them, the heat-shock 70-kD protein and S100 calcium-binding protein A6 were down-regulated proteins; thioredoxin, glyceraldehyde-3-phosphate dehydrogenase, and beta-actin were up-regulated proteins. The function of the five proteins were closely associated with the growth, apoptosis, santioxidant and ignal transduction of MARC-145 cells. 【Conclusion】The quantity of functional proteins of the MARC-145 was changed which were infected by highly pathogenic PRRSV.

Expression, Sequence Analysis and Immunohistochemical Localization of Mitf-M Transcription Factor in Alpaca Skin

ZHU Zhi-Wei, HE Jun-Ping, YU Xiu-Ju, CHENG Zhi-Xue, DONG Chang-Sheng

Scientia Agricultura Sinica. 2012, 45(4): 794-800. doi:10.3864/j.issn.0578-1752.2012.04.021

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【Objective】 Cloning, sequence analysis and tissue location of the Mitf-M CDS domain sequence of alpaca skin could provide a foundation for studying coat color expression mechanism.【Method】The CDS domain sequence of Mitf-M was amplified by RT-PCR, multiple sequence alignment of the amino acid sequence was analyzed by Clustal X and BioEdit software. The distribution of Mitf-M protein in alpaca skin was studied by immunohistochemistry.【Result】Mitf-M CDS domain of alpaca is composed by 419 amino acids, with bHLH-zip conserved domain of transcription factor family. Phylogenetic analysis showed that Mitf-M had the highest homology with cattle (Bos Taurus) and dogs (Canis familiaris). The Mitf-M protein was mainly located in melancytes during the basal cells of hair bulb and around dermis papilla. 【Conclusion】 Compared with other species, Mitf-M in alpaca skin are highly conservative, indicating that Mitf-M plays an important role in the transcriptional regulation and pigmentation.

Characteristics of Dry Matter Accumulation and Translocation During the Wheat Genetic Improvement and Their Relationship to Grain Yield

TIAN Zhong-Wei, WANG Fang-Rui, DAI Ting-Bo, CAI Jian, JIANG Dong, CAO Wei-Xing

Scientia Agricultura Sinica. 2012, 45(4): 801-808. doi:10.3864/j.issn.0578-1752.2012.04.022

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【Objective】To verify the characteristics of dry matter accumulation, translocation and their relationships to grain yield during wheat genetic improvement will facilitate development of new cultivars for stable and high yielding wheat production.【Method】 Thirty two wheat cultivars bred or widely planted in China from 1950 to 2005 were grown in field experiments from 2007 to 2009. Dry matter accumulation and growth rate at different stages, leaf area index (LAI) and net photosynthetic rate (Pn) were measured, and dry matter translocation and their relationship to grain yield formation were analyzed. 【Result】 Grain yield and harvest index increased linearly with the progress of wheat cultivar evolution, whereas the biomass of the cultivars from 1950 to 1960 reduced and thereafter maintained stable. Leaf area, LAI and flag leaf Pn at anthesis increased linearly with the cultivars evolution, supplying the sources of dry matter and energy for yield formation after anthesis. Genetic improvement increased the dry matter accumulation and growth rate at the growth stages of emergence to jointing and anthesis to maturity whereas reduced the contribution of post-anthesis accumulated dry matter to grains (CPA). Grain yield was significantly and positively associated with the contribution of pre-anthesis translocation to grains (CPT), dry matter accumulation and growth rate from emergence to jointing and anthesis to maturity, whereas significantly and negatively correlated with CPA, dry matter accumulation and growth rate from jointing to anthesis.【Conclusion】Dry matter production capacity and production efficiency were improved during wheat cultivar evolution, yet dry matter accumulation at different growth stages were coordinated and the contribution of pre-anthesis and post-anthesis accumulation to grains became more balance. Improving the pre-jointing vegetative growth post-anthesis accumulation and CPT are important physiological basis for wheat grain yield enhancement, which will be a key improvement target for high yield wheat breeding in the future.

Comparison of Profiling Rice Volatiles by Means of Solid Phase Microextraction

SU Yi-Juan, WANG Rui-Long, YE Mao, SONG Yuan-Yuan, ZENG Ren-Sen

Scientia Agricultura Sinica. 2012, 45(4): 809-814. doi:10.3864/j.issn.0578-1752.2012.04.023

Abstract ( 1010 )

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【Objective】The objective of the study is to provide appropriate volatile collection methods of solid phase microextraction (SPME) for analyzing induced volatiles from plants through comparison of difference in rice volatile profiles collected by different fibres. 【Method】 Three SPME fibres including 100 µm polydimethylsiloxane (PDMS), 7 µm PDMS and 85 µm polyacrylate（PA) were used to extract volatiles emitted from rice plants that were treated with methyl jasmonate. Gas chromatography (GC) and gas chromatography-mass spectrometry (GC-MS) were used to compare the difference in rice induced volatiles in terms of their amount, profile and composition of extracted volatiles. The difference between SPME method and dynamic headspace sampling (DHS) technique was investigated. 【Result】 Under the same condition, the maximum amount and variety of volatiles were found in rice volatiles extracted by 100 µm PDMS, the minimum amount and variety by 7 µm PDMS. The composition of volatiles obtained by SPME and DHS was similar, but profile of volatile compounds differed from each other. 【Conclusion】 Three fibres showed varied efficiency and selectivity in extracting volatile compounds. SPME fibres should be chosen appropriately according to the property of target compounds in plant volatiles. Both SPME and DHS have their own advantages and disadvantages in plant volatiles extraction and analysis.

Cloning and Analysis of Three Ethylene Response Factor Genes in Alfalfa（Medicago sativa L.）

CHEN Ting-Ting, YANG Qing-Chuan, KANG Jun-Mei, DING Wang, ZHANG Tie-Jun, ZHANG Xin-Quan

Scientia Agricultura Sinica. 2012, 45(4): 815-822. doi:10.3864/j.issn.0578-1752.2012.04.024

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【Objective】 Cloning and characterization of three ethylene response factor genes in alfalfa to provide a foundation for further study of their function.【Method】Three ethylene response factor genes, MsERF5, MsERF8, and MsERF11 were isolated from alfalfa by PCR, and their expression patterns under different treatments and in different tissues were analyzed using semi-quantitative RT-PCR. The expression vectors of these three genes, was constructed, and then were introduced into tobacco by Agrobacterium mediated transformation system and the salt-tolerance of trangenic plants was analyzed.【Result】 MsERF5, MsERF8, and MsERF11 had no intron. The expression patterns of these three genes were significantly different in different tissues, the expression level of MsERF5 in roots, leaves and flower buds was higher than in other tissues. The expression of MsERF8 was higher in roots and leaves, and the expression of MsERF11 in leaves was higher than in other tissues, the expression of MsERF5, MsERF8, and MsERF11 were both induced by salt, drought, Al3+ and diverse hormones. The expression vectors of these three genes were constructed, and they were introduced into tobacco by Agrobacterium mediated transformation system. The calluses transformed MsERF5, MsERF8, and MsERF11 were changed into adventitious bud differ from the non-transformed calluses. The chlorophyll and soluble sugars contents of the transgenic plants were significantly higher than wild-type plants. The electrolyte leakage was significantly lower than wild-type plants. 【Conclusion】MsERF5, MsERF8, and MsERF11 had no intron, the expression patterns of these three genes were significantly different in different tissues, and both of them could be induced by abiotic stresses and diverse hormones treatments. All of them could change the calluses into adventitious buds on the saline medium and produce transgenic plants, and the salt-tolerance of transgenic plants was higher than wild-type plants.

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