Scientia Agricultura Sinica

Studies on Mapping and Inheritance of Resistance Genes to SMV Strain SC10 in Soybean

LI Chun-Yan, YANG Yong-Qing, WANG Da-Gang, LI Hua-Wei, ZHENG Gui-Jie, WANG Tao, ZHI Hai-Jian

Scientia Agricultura Sinica. 2012, 45(21): 4335-4342. doi:10.3864/j.issn.0578-1752.2012.21.001

Abstract ( 635 )

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【Objective】The inheritance of resistance and allelism of resistance genes to SMV strain SC10 was studied and the resistance genes to SC10 in Kefeng No.1 were mapped.【Method】The resistant soybean cultivars to SMV strain SC10(Kefeng No.1, Jinda74, Dabaima, Fendou56, Zhongzuo229, Xudou No.1, Pixianchadou, Kwanggyo and Yuejin No.4)were crossed with susceptible (S) cultivars (Nannong 1138-2 or 8101), respectively, to determine the inheritance of resistance to SC10. Each R parents were also crossed with each other to test the allelism of the resistance genes. The bulked segregant analysis (BSA) of SSR markers and F2 population from Kefeng No.1 × Nannong 1138-2 were used in mapping the resistance gene.【Result】The F1 of crosses from resistant cultivars (Kefeng No.1, Jinda74, Dabaima, Fendou56, Zhongzuo229, Xudou No.1) and susceptible (S) cultivars (Nannong 1138-2 or 8101) were resistant, F2 segregated in 3 resistant﹕1 susceptible ratio, and 1R﹕2Seg﹕1S in F2:3 families. The F1 and F2 populations from Jinda74×Fendou56 and Xudou No.1×Pixianchadou exhibited complete resistant. F1 from the two crosses of Kefeng No.1×Kwanggyo, Fendou56 and Zhongzuo229, Jinda74×Zhongzuo229 and Kwanggyo, Dabaima×Fendou56 and Kefeng No.1. Yuejin No.4×Kwanggyo were all resistant while their correspond F2 population segregated in a ratio of 15(R)﹕1(S). The seven SSR markers (Satt558, Sat_254, Satt634, Gm020580, Gm020584, Gm020562 and Gm020546) were found linking to RSC10. The genetic distance between the resistance gene RSC10 and SSR markes were 6.1, 2.0, 0.9, 0.8, 2.0, 4.6, and 9.2 cM, respectively.【Conclusion】 Kefeng No.1, Jinda74, Dabaima, Xudou No.1, Fendou 56, Zhongzuo229, respectively, have a single dominant gene controlling the resistance to SC10. The resistance genes in Jinda74 and Fendou 56, Xudou No.1 and Pixianchadou were at a same locus or very closely linked. It was inferred that the resistance genes between Kefeng No.1 and Kwanggyo, Fendou56, Zhongzuo229; between Jinda74 and Zhongzuo229, Kwanggyo; between Dabaima and Fendou56, Kefeng No.1; between Yuejin No.4 and Kwanggyo were not at the same locus. The resistance gene carried by Kefeng No.1 to SC10 was located on D1b linkage group.

Arabidopsis TUA2 Gene Mediates Seed Germination Under ABA Stress

LIU Hai-Hao, WU Li-Zhu, YUE Zhi-Liang, PAN Yan-Yun

Scientia Agricultura Sinica. 2012, 45(21): 4343-4350. doi:10.3864/j.issn.0578-1752.2012.21.002

Abstract ( 676 )

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【Objective】This study would characterize the TUA2 gene involving in seed germination under ABA stress in Arabidopsis by the method of reverse genetics. 【Method】 The Arabidopsis T-DNA inserted into TUA2 gene mutants were obtained from ABRC. The T-DNA insertion sites of mutants were analyzed by Tail PCR. Over-expression constructs of TUA2 were introduced into Arabidopsis by an Agrobacterium-mediated method. RT-PCR was used to detect TUA2 and ABA-responsive genes accumulation in mutants and transgenic plants. The seed germination of transgenic and mutant lines was tested on MS medium with different concentrations of ABA. 【Result】 The T-DNA was inserted into the gene promoter region in mutants tua2-1 and tua2-2, and led to the increasing accumulation of the TUA2. In the medium of MS/0.8 μmol•L-1 ABA, the seed germination rate of wild type was 76% after five days, whereas all the TUA2 over-expression lines, mutants and transgenic plants, were 6%-18%. The expression of ABII, HAB and P5CS was induced by ABA in wild type, but not in mutants and transgenics. 【Conclusion】 The T-DNA insertion of the mutants tua2-1 and tua2-2 resulted in the increasing expression of TUA2 gene. The TUA2 gene might be involved in plant responses to ABA by mediating the transcription of ABA-responsive genes.

Microtubules in Guard Cells Involve in Serine/Threonine Phosphorylation During Stomatal Movement

YE Zi, HUANG Cong-Cong, YU Rong

Scientia Agricultura Sinica. 2012, 45(21): 4351-4360. doi:10.3864/j.issn.0578-1752.2012.21.003

Abstract ( 727 )

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【Objective】 The interaction between microtubules (MTs) and protein serine/threonine phosphorylation in guard cells was investigated in order to provide a reliable evidence for better understanding of the mechanism of stomatal movment. 【Method】Pharmacological analysis, confocal laser scanning microscopy, and western blotting were used to explore the possible interaction of these two elements. 【Result】 In vivo stomatal aperture assays revealed that staurosporine (STS), one well-known specific inhibitor of serine/threonine protein kinase, enhanced the light-induced stomatal opening. While, okadaic acid (OA) and calyculin A (CalA), another two inhibitors of 1/2A phosphatase, showed the opposite effects under the same condition. In addition, it was found that these changes of stomatal movement could be reversed or became intensified to some extent when microtubule-specific drug, taxol or vinblastine was present during the treatments above. Furthermore, OA and CalA also had significant influences on the MT organization in the opening guard cells. In the presence of OA or CalA, the fine radial MT arrays tended to be broken down or rearranged into random network. And guard cells kept in STS showed increase in radial MT arrays and stomatal aperture. MT-targeted drugs, taxol and vinblastine, also altered the effects of three inhibitors (OA, CalA, STS) on MT organization partiallly in a similar manner. Immunoblot detection of total extract from guard cells confirmed that tubulin-subunits were phosphorylated on serine. 【Conclusion】 The results suggest that reversible protein serine/threonine phosphorylation may regulate the MT dynamics of guard cells during stomatal movement.

Dynamic Simulation on Angle Between Stem and Sheath in Different Rice Cultivars and Nitrogen Rates

ZHANG Yong-Hui, TANG Liang, LIU Xiao-Jun, CAO Wei-Xing, ZHU Yan

Scientia Agricultura Sinica. 2012, 45(21): 4361-4368. doi:10.3864/j.issn.0578-1752.2012.21.004

Abstract ( 744 )

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【Objective】The objective of this paper is to develop a dynamic simulation model on angle between stem and sheath at different leaf positions on main stem in different rice cultivars and nitrogen application rates.【Method】Based on rice experiments with different cultivars and nitrogen rates, the values of angel between stem and sheath of different leaf positions on main stem during different development stages were continuously observed, a dynamic simulation model for changes of angle between sheath and stem at different leaf positions on main stem was developed by using dynamic modeling technology. 【Result】 Rice angle between stem and sheath increased with leaf development progress, the duration from the beginning of angle formation to the maximal value was about three leaf ages and the angles increased with the increasing nitrogen rates. From the first leaf to the third leaf, the maximum angle between stem and sheath at each leaf position increased with the increasing leaf position, and then decreased with the increase of leaf position. The logistic equation could be used to describe the change process of the angle between stem and sheath on main stem with growing degree days (GDD) in different varieties and nitrogen rates, and the maximum angle between stem and sheath with leaf position on main stem could be described with linear piecewise function. A variety parameter (the maximum angle between stem and sheath at the third leaf position) was introduced to quantify the variety characteristics on the angle between stem and sheath. Nitrogen factor was also used to reflect the effect of nitrogen on the angle between stem and sheath. 【Conclusion】A set of independent pot and pool experiments data were used to test the model, and the root mean square error (RMSE) between observed and simulated values was 2.31° and 2.78°, respectively, and the relative RMSE (RRMSE) was 11.56% and 14.77%, respectively, which indicated that the model has a good performance on predicting the angle between stem and sheath at different leaf positions on main stem. These results would provide technological support for the visualization of rice plant.

Change Detection on Wetlands Using High Spatial Resolution Imagery

ZHU Jin-Xia, GUO Qing-Hua, WANG Ke

Scientia Agricultura Sinica. 2012, 45(21): 4369-4376. doi:10.3864/j.issn.0578-1752.2012.21.005

Abstract ( 650 )

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【Objective】With respect to the change detection on wetlands, very high spatial resolution images of drained managed wetland ponds were used, which could provide more information for further management. 【Method】 The proposed method is based on pixel-oriented difference image and object-based post-classification(OB-M). Multivariate alteration detection (MAD) transformation was used to get the extended difference image, and object-based decision tree classification was applied on MAD components to detect the true change information of difference image, which had a very significant shape feature.【Result】 The proposed OB-MAD can successfully detect the false change information, such as the inevitable mis-registration, shadow and vegetation phenology differences. Compared with the traditional MAD method with thresholds (Threshold-MAD) and the traditional object-based post-classification method (OB-T), the proposed OB-M method produced the highest accuracy, which took advantage of both pixel- and object-based technology.【Conclusion】Results indicated that the object-based post-classification on MAD components can well detect the change information of wetlands.

Analysis of Race and Genetic Relationship of Cercospora sojina in Heilongjiang Province

DING Jun-Jie, GU Xin, YANG Xiao-He, ZHAO Hai-Hong, SHEN Hong-Bo, JIANG Cui-Lan, SHI Xiang-Lin, LIU Chun-Yan, HU Guo-Hua, CHEN Qing-Shan

Scientia Agricultura Sinica. 2012, 45(21): 4377-4387. doi:10.3864/j.issn.0578-1752.2012.21.006

Abstract ( 770 )

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【Objective】The objective of this study is to survey race types of Cercospora sojina and to analyze the genetic relationship of races of C. sojina in Heilongjiang Province. 【Method】Samples of C. sojina were collected and pathogens were isolated in Heilongjiang Province from 2006 to 2011. Twenty-four strains of C. sojina were analyzed by EST-SSR with 11 pairs SSR primers. 【Result】Fifteen races were identified from 2 674 strains of C. sojina obtained in this study. The results of SSR showed that the amount of the allelic variation in all strains was 46, and the average was 4.2. Similarity index between strains ranged from 0.091 to 0.956, and the average was 0.589. The cluster results had the high relativity to race of C. sojina strains.【Conclusion】There was a rich genetic diversity among C. sojina in Heilongjiang Province. The genetic relationship of C. sojina is closely related to race type, and is irrelevant to collective time and place.

Rapid Molecular Diagnosis Based on SCAR Marker System for Cereal Cyst Nematode

QI Xiao-Li, PENG De-Liang, PENG Huan, LONG Hai-Bo, HUANG Wen-Kun, HE Wen-Ting

Scientia Agricultura Sinica. 2012, 45(21): 4388-4395. doi:10.3864/j.issn.0578-1752.2012.21.007

Abstract ( 732 )

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【Objective】 The objective of this study is to build a SCAR marker system for rapid molecular detection of cereal cyst nematode (Heterodera avenae) from mixtures of different nematodes. 【Method】 With the method of RAPD and SCAR, a single-step PCR was used to specially detect H. avenae. 【Result】 Using the SCAR marker system established in this study, the target fragment was amplified from the DNA templates of H. avenae only, from other 8 species including 32 populations. The SCAR marker system could detect cysts and the J2 of H. avenae, and the specific fragment could be clearly identified when the dilution was 1/2000 of a cyst or 1/80 of a J2 for all replicates. 【Conclusion】 The establishment of the SCAR marker system for rapid molecular detection of H. avenae can be used to detect the H. avenae from the mixed nematode samples rapidly, and the detection is accurate and highly sensitive.

Kinetics of K+ Adsorption and Desorption of Brown Soil Under Long-Term Fertilization

LI Na, HAN Li-Si, WU Zheng-Chao, WANG Yue, LAN Yu, HAN Xiao-Ri

Scientia Agricultura Sinica. 2012, 45(21): 4396-4402. doi:10.3864/j.issn.0578-1752.2012.21.008

Abstract ( 721 )

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【Objective】The objective of study was to investigate the kinetics characteristics of K+ adsorption and desorption of brown soil under long-term (32 years) fertilization.【Method】Nine samples of different treatments were flowed through with deionized water as pretreatment. Miscible displacement techniques were used to study the adsorbing-desorbing process. 【Result】The equilibrium time of adsorption ranged from 26 min to 65 min, and that of desorption ranged 65-130 min. The ranges of equilibrium adsorbing capacity and the equilibrium desorbing capacity were 7.40-19.44 cmol•kg-1 and 0.070-0.258 cmol•kg-1. The relation between velocity of potassium adsorbing and time could be described with the model of V=A+Blnt. There were correlation between parameters of adsorption and desorption were related with CEC, Clay, OM and available K in different degrees. First-order, Elovich, Parabolic diffusion and Dual constant were used to describe the kinetics of K adsorption and desorption of experimental soils. The results showed that Elovich and First-order were the best kinetic models for describing adsorption and desorption, respectively.【Conclusion】There were great different characteristics of adsorption and desorption under different fertilizations. The ability of adsorption could be increased by applying organic manure. And the ability of desorption improved with potassium rates. The effect of long-term fertilization on K+ adsorption and desorption kinetics of brown soil was up to the changes of physic-chemical properties of soil. There were different dynamic processes of potassium in soil under different fertilizations.

Effects of Combined Application of Organic Manure and Chemical Fertilizers on Ammonia Volatilization from Greenhouse Vegetable Soil

HAO Xiao-Yu, GAO Wei, WANG Yu-Jun, JIN Ji-Yun, HUANG Shao-Wen, TANG Ji-Wei, ZHANG Zhi-Qiang

Scientia Agricultura Sinica. 2012, 45(21): 4403-4414. doi:10.3864/j.issn.0578-1752.2012.21.009

Abstract ( 728 )

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【Objective】Excessive use of chemical nitrogen fertilizers suffer from nitrogen loss through ammonia (NH3) volatilization, resulting in decreasing effectiveness of the fertilizers and increasing the risk of environmental pollution. Less information is available on the effect of different fertilization patterns on NH3 volatilization from greenhouse vegetable soils. Based on balanced fertilization and optimized irrigation, this study was conducted to determine the effect of combined application of organic manure and chemical fertilizers on NH3 volatilization. 【Method】A field experiment was designed to study the dynamics of NH3 volatilization by the venting method under celery-tomato rotation system in Tianjin from September 2010 to June 2011. The field experiment consisted of six treatments of 1 (No N), 2 (4/4 CN+0/4 PN, all nitrogen in chemical fertilizer, PN denotes nitrogen in pig manure), 3 (3/4 CN+1/4 PN), 4 (2/4 CN+2/4 PN), 5 (2/4 CN+2/4 SN, SN denotes nitrogen in straw), and 6 (CF, conventional fertilization). Equal NPK was designed for treatments 2, 3, 4, and 5. Rates of P2O5 and K2O application were respectively equal for each of treatments 1, 2, 3, 4 and 5. 【Result】 The peak of NH3 volatilization rate occurred within 2-3 days after basal fertilization, whereas in first day after fertilizer top-dressing (fertilization and irrigation were carried out simultaneously) during the celery and tomato growth period. The NH3 volatilization rate from the nitrogen application treatment closed to the rate from the no nitrogen application treatment until first 8-10 days after basal fertilization and first 10-11 days after fertilizer top-dressing, respectively. 70%-80% of total NH3 volatilization during celery or tomato growth period were from over the first 10 days after basal fertilization to the first 11 days after the second fertilizer top-dressing. Distinct total NH3 volatilization during tomato growth period with higher temperature was found, with the total amounts of NH3 volatilization for all treatments being 3.0 times as high as that of the celery growth period. Compared with the conventional fertilization with much higher rates of N fertilizer, the combined application patterns of organic manure and chemical fertilizers significantly decreased the total NH3 volatilization by 50.0% and 49.6% during the celery and tomato growth period, respectively. The NH3 volatilization gradually decreased with the increase of the organic nitrogen rates. Straw application compared with pig manure application with the same N rate respectively decreased the NH3 volatilization by 32.4% and 30.0% during the growth period of celery and tomato under this experimental condition. The proper pattern of combined application of organic manure and chemical fertilizers based on yield, income and environment was the pattern of combined application of 3/4 N in chemical fertilizer and 1/4 N in organic manure under this experimental condition. 【Conclusion】The pattern of combined application of organic manure and chemical fertilizers significantly decreased the total NH3 volatilization, with the fertilization pattern being economic benefit-significantly, feasibility-strongly and environment-friendly, and its application in sustainable greenhouse vegetable production is prospective.

Effects of NaCl Stress on Root Exudates of Cucumber Cultivars

WU Feng-Zhi, ZHOU Xin-Gang, BAO Jing

Scientia Agricultura Sinica. 2012, 45(21): 4415-4427. doi:10.3864/j.issn.0578-1752.2012.21.010

Abstract ( 761 )

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【Objective】This study aimed to elucidate the relationship between NaCl stress and root exudates components. 【Method】 Cucumber cultivars with different NaCl tolerances were used as experimental materials. Root exudates components in the whole growth period of cucumber under different NaCl concentrations (0, 100, 200, 300 mmol NaCl•L-1) were analyzed. 【Result】Except for the 300 mmol NaCl•L-1 concentration treatment, soluble sugar content in NaCl-tolerant cucumber root exudates was higher than that in NaCl-sensitive cultivar on 50 d after cucumber transplanting. Sixteen kinds of free amino acids were detected in cucumber root exudates. The variety and contents of free amino acid were higher in NaCl-tolerant cucumber root exudates than in NaCl-sensitive cultivar at the seedlings stage. On 30 d after cucumber transplanting, total amino acid content in NaCl-sensitive cultivar under all concentrations of NaCl and in NaCl-tolerant cultivar under 100 and 200 mmol NaCl•L-1 treatments were lower than that in the control. On 50 d after transplanting, the variety and contents of free amino acid were higher than that on 30 d after transplanting; total amino acids content in NaCl-sensitive cultivar were higher than that in NaCl-tolerant cultivar under 200 and 300 mmol NaCl•L-1 treatments, whereas the trend was the contrary for other NaCl concentrations. On 70 d after transplanting, the variety and contents of free amino acid were higher than that on 30 and 50 d after transplanting, except for the 300 mmol NaCl•L-1 treatment, total amino acids content in NaCl-sensitive cultivar were higher than in NaCl-tolerant cultivar. Under the same NaCl concentration, threonine, serine and glycine contents in the NaCl-sensitive cultivar were higher than in the NaCl-tolerant cultivar, whereas cysteine content was the contrary. There were 65 kinds of compounds were detected in cucumber root exudates with GC-MS method. The variety of these compounds was higher on 30 d after transplanting than on 50 and 70 d after transplanting. The total content of these compounds was higher on 50 d after transplanting than on 30 and 70 d after transplanting. In 300 mmol NaCl•L-1 treatment, the total content of these compounds was higher in NaCl-sensitive cultivar than in NaCl-tolerant cultivar on 30, 50 and 70 d after transplanting. 【Conclusion】Soluble sugar content in cucumber root exudates was affected by NaCl concentration and cucumber growth stage. Soluble sugar content in NaCl-sensitive cultivar was higher than in NaCl-tolerant cultivar. NaCl stress increased the variety of amino acids. The variety and total content of compounds, as detected by GC-MS, had the trend to decrease with the increass of NaCl concentration and cucumber development.

Accumulation of Flavanols, Expression of Leucoanthocyanidin Reductase Induced by UV-C Irradiation During Grape Berry Development

WEN Peng-Fei, XING Yan-Fu, NIU Tie-Quan, GAO Mei-Ying, NIU Xing-Yan

Scientia Agricultura Sinica. 2012, 45(21): 4428-4436. doi:10.3864/j.issn.0578-1752.2012.21.011

Abstract ( 599 )

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【Objective】 The effect of ultraviolet C on the accumulation of flavanols in grape berry during its development was studied.【Method】The 5-year old grapevine (Vitis vinifera L. cv. Cabernet Sauvignon) was subjected to the regular UV-C irradiation during berry development, and the accumulation of flavanols, the enzyme activity of leucoanthocyanidin reductase (LAR), the transcriptional level and translation level of Vv lar1 and lar2 were determined by spectrophotometer, real time PCR, and Western Blot.【Result】 The accumulation pattern of flavanols in grape berry during its development was not changed, whereas the accumulation of flavanols was promoted by UV-C irradiation, especially in the young berry. In addition, the activity of LAR, the key enzyme in the flavanols biosynthetic pathway, was induced by the UV-C irradiation. The synthesis of LAR1 and LAR2 new protein and the accumulation of Vv lar1 and lar2 mRNA induced by UV-C irradiation were also observed. 【Conclusion】 Results of the study indicated that the UV-C irradiation could induce the transcription of Vv lar1 and Vv lar2, the synthesis of LAR1 and LAR2 new protein, the increase of LAR activity, and resulted in the accumulation of flavanols in berry.

Changes of Polar Metabolites in Guoqing No.1 Satsuma Mandarine During Fruit Ripening

LIU Shu-Zhen, HAN Jing-Wen, YUN Ze, WANG Jin-Qiu, XU Juan, ZHANG Hong-Yan, DENG Xiu-Xin, CHENG Yun-Jiang

Scientia Agricultura Sinica. 2012, 45(21): 4437-4446. doi:10.3864/j.issn.0578-1752.2012.21.012

Abstract ( 681 )

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【Objective】 To provide novel objective evaluation methods for the judgment of fruit maturation and its internal quality, the changes of polar metabolites in Guoqing No.1 satsuma mandarin (Citrus unshiu Marc. cv.Guoqing No.1) was investigated during fruit ripening. 【Method】 Extraction and analysis methods of citrus primary metabolites were established by using gas chromatography-mass spectrometry (GC-MS). Based on the method, polar metabolites in Guoqing No.1 satsuma mandarin at three different ripening stages (mature green, color-change and maturation stage) were assayed, and principal component analysis (PCA) was employed to explore the polar metabolite profiles.【Result】A total of 37 kinds of metabolites were identified. As the fruits turned orange, the levels of fructose, mannose and most amino acids (especially GABA and proline) in flavedo increased significantly, meanwhile, citric acid and some fatty acids decreased gradually. However, fewer metabolites changed in pulp during the processes. The contents of mannose, arabinose, oxalic acid, phosphoric acid, succinic acid, 2-keto-glutaric acid, 9,12-octadecadienoic acid, octadecanoic acid, palmitic acid and tetradecanoic acid reached the highest level at mature green stage and subsequent decreased gradually, while the content of sucrose, GABA and a variety of organic acids were largely accumulated. PCA results showed that polar metabolites in different tissues and ripening stages were separated very well in two dimensions.【Conclusion】 The results demonstrated that, with orange color enhancement, polar metabolites in citrus fruit changed dramatically. Moreover, the changes of these polar metabolites, especially sugars and organic acids, showed obvious tissue specificity, and some polar metabolites could be regarded as standards for citrus maturity index.

Effects of High Temperature-Short Time Steam Blanching (HTSTSB) on Myrosinase Activity and Nutrients of Maca

TU Xing-Hao, ZHENG Hua, ZHANG Hong, GAN Jin, ZHANG Wen-Wen, LI Kun, XU Juan

Scientia Agricultura Sinica. 2012, 45(21): 4447-4456. doi:10.3864/j.issn.0578-1752.2012.21.013

Abstract ( 832 )

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【Objective】 In order to obtain more glucosinolates and improve the quality of the maca after storage and processing, the application of high temperature-short time steam blanching (HTSTSB) on myrosinase inactivation was investigated. 【Method】Three main factors, which would influence the myrosinase activity and glucosinolates content of maca, such as steam pressure, steam processing time and material capacities, were considered in the phase of single factor tests. Based on the result, the response surface methodology (RSM) test was designed. 【Result】With the comprehensive evaluation and analysis of effects of myrosinase inactivation and glucosinolates preservation, the optimum conditions were obtained as follows: steam pressure was 0.35 MPa, steam processing time was 50 s, and the material capacity was 240 g. Under the optimal conditions, the myrosinase activity of maca was lowered to 1.96%, and the retained content of glucosinolates could reach to 0.98%. Nutrients were retained more by HTSTSB compared to hot water blanching and microwave blanching. 【Conclusion】 The myrosinase activity of maca could be effectively passivated by HTSTSB, and sensory quality was better maintained after treatment. Compared with conventional blanching, HTSTSB has a clear advantage.

Study on Optimization of Non-Starch Polysaccharide Enzymes of Broiler Diets in Vitro

HE Ke-Lin, SA Ren-Na, GAO Jie, LI Dong-Wei, ZHUANG Xiao-Feng, ZHANG Hong-Fu

Scientia Agricultura Sinica. 2012, 45(21): 4457-4464. doi:10.3864/j.issn.0578-1752.2012.21.014

Abstract ( 789 )

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【Objective】The study was conducted to discuss the effects of non-starch polysaccharides (NSP) enzymes on the in vitro digestive energy (IVDE) of broiler diets of different contents of NSP.【Method】The first step was to investigate the dose-response of the NSP enzymes on the IVDE in diets using one-way randomized experimental design. Five NSP enzymes including xylanase, β-glucanase, cellulose, pectinase, and β-mannanase were added into corn-soybean diet and wheat-soybean diet, from 0 to 900 µg•g-1. The second step was to screen the optimum enzyme combinations using quadratic regress-orthogonal rotary design. The method of testing IVDE was a two-stage enzymic hydrolysis with pepsin-pancreatin in vitro.【Result】The IVDE of broiler diet regulating by NSP enzyme appeared S state diagram. The xylanase had more promotion effects than other NSP enzymes. It increased IVDE by 0.24 MJ•kg-1 and 0.40 MJ•kg-1 in corn-soybean diet and wheat-soybean diet, respectively. In corn-soybean diet, the optimum enzyme combinations were xylanase 34 836.4 U•kg-1, β-glucanase 6 762.0 U•kg-1, cellulose 1 159.2 U•kg-1, pectase 872.6 U•kg-1, and β-mannanase 24 535.9 U•kg-1. In wheat-soybean diet, the optimum enzyme combinations were xylanase 65 405.5 U•kg-1, β-glucanase 10 131.5 U•kg-1, cellulose 980.0 U•kg-1, pectase 501.6 U•kg-1, and β-mannanase 5 141.4 U•kg-1. 【Conclusion】The findings showed that using five kinds of NSP enzymes in corn-soybean and wheat-soybean diets of broiler could improve the in vitro digestive energy.

Cloning, Expression and Bioinformatics Analysis of Porcine CatSperB and CatSperG Genes

SONG Cheng-Yi, ZHOU Jia-Qing, FENG Xiao-Jun, XIE Yu-Xiu, LI Qing-Ping, WU Han, GAO Bo, WANG Xiao-Yan

Scientia Agricultura Sinica. 2012, 45(21): 4465-4474. doi:10.3864/j.issn.0578-1752.2012.21.015

Abstract ( 638 )

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【Objective】The aim of the current study is to confirm the existence of porcine CatSperB and CatSperG genes, and investigate the protein structures, evolutionary relationship and the spatial-temporal expression profiles of CatSperB and CatSperG. 【Method】The in silico and molecular cloning was used to identify the full length cDNAs of porcine CatSperB and CatSperG, and the spatial-temporal expression profile was investigated by qualitative and fluorescence quantitative RT-PCR. 【Result】The in silico transcripts of 3 508 bp CatSperB and 3 715 bp CatSperG were identified, and they contain 3 330 bp and 3 483 bp ORFs of CatSperB and CatSperG, respectively, and the sequences were confirmed by TA cloning. The sequence similarity of coding sequences (CDS) of porcine CatSperB and CatSperG with human, cattle, horse, and dog, and other animals was above 80%. CatSperB is a 125.79 kD and stable protein, while CatSperG is a 133.40 kD and unstable protein. Both CatSperB and CatSperG contain seven conservative trans-membrane domains, and a coiled-coil motif was also identified in the C terminal of CatSperG, but this motif was not found in CatSperB. The porcine CatSperB and CatSperG displayed higher degree of homology with the orthologs of cattle, dog and horse, and lower homology with those of human and mouse. The RT-PCR analysis showed that CatSperB and CatSperG were detected mainly in testis, but CatSperB also expressed in other tissues. The mRNA expression of CatSperB and CatSperG was significantly improved at the around stage of spermatogenesis (Day 60), puberty (Day 90) and sex maturity (Day 150) (P＜0.05). 【Conclusion】The cDNA clones of CatSperB and CatSperG, and a series of bioinformatics parameters of these proteins were obtained. Seven conservative trans-membrane domains in CatSperB and CatSperG and the homology among species were revealed. Furthermore, results of this study had proved that CatSperB and CatSperG expressed mainly in tests, and the change of mRNA expression was paralleled with sexual development of boar.

Characterization of Tissue Expression and Cloning of Nuclear Factor I/B Gene in Chinese Merino (Xinjiang Junken Type)

RONG 恩Guang, YU Lei, YANG Hua, ZHANG Yong-Sheng, MA Chun-Ping, LI Hui, WANG Ning

Scientia Agricultura Sinica. 2012, 45(21): 4475-4483. doi:10.3864/j.issn.0578-1752.2012.21.016

Abstract ( 705 )

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【Objective】The objective of this study was to explore the expression level of nuclear factor I/B gene in various sheep tissues, and clone the full-length coding sequence (CDS) of sheep NFIB gene in Chinese Merino (Xinjiang Junken type).【Method】Tissue expression profile of NFIB gene was detected by semi-quantitative reverse transcription PCR (SqRT-PCR). The full-length CDS region of NFIB gene was amplified by RT-PCR, cloned into T vector, and sequenced. The sequence was analyzed by using bioinformatics software.【Result】SqRT-PCR analysis showed that the NFIB gene was ubiquitously expressed in sheep internal organs and skin with different degrees. Comparatively, it was highly expressed in skin. The skin expression of NFIB gene in super fine wool strain had significant difference in different seasons (P＜0.05), but no significant difference in skins from various anatomic locations. There was no significant difference in the NFIB gene expression of body side skin among the 6 sheep strains/breeds (P＞0.05). Sequence analysis indicated that the sheep NFIB gene generated three NFIB protein isoforms of 420, 375 and 345 aa by alternative splicing.【Conclusion】NFIB gene is highly expressed in the skin of Chinese Merino (Xinjiang Junken type), and its expression in the skin of super fine wool strain has significant difference in different seasons. Sheep NFIB has at least three protein isoforms. This study not only has laid a foundation for clarifying the function of sheep NFIB gene, but also provided a theoretical basis for creating high quality merino sheep.

Genetic Diversity of Northeastern Black Bee (Apis mellifera ssp.) in China Based on Microsatellite Markers

PENG Wen-Jun, LUO Qi-Hua, LI Chang-Chun, LIU Xiao-Dong, MA Yu-Zhen, GUO Jun, AN Jian-Dong

Scientia Agricultura Sinica. 2012, 45(21): 4484-4491. doi:10.3864/j.issn.0578-1752.2012.21.017

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【Objective】 The objective of the study is to discuss the genetic diversity of the northeastern black bee (Apis mellifera ssp., also named Raohe black bee) from China National Nature Reserve for the Black Bee (CNNRBB). 【Method】The genetic diversity of northeastern black bee from 7 districts in Raohe County, Heilongjiang Province were surveyed by using 16 microsatellite markers. 【Result】A total of 725 alleles were detected at 16 loci among 245 workers, all loci except A028, A024 and A088 showed high polymorphism in the populations studied, the average polymorphism information content (PIC) was 0.60±0.05, the mean observed heterozygote (Ho) was 0.62±0.21, and the expected heterozygote (He) was 0.65±0.19, positive values of the inbreeding coefficient (Fis) were found (excluding two populations), indicating a certain level of inbreeding. Coefficients of genetic differentiation (Fst) between populations ranged from 0.03 to 0.14, and the Nei's genetic distance (DA) was from 0.08 to 0.35, moderate genetic differentiation (Fst =0.14) and a high estimate of gene flow (Nm=1.59) were detected in the total populations. The clustering analysis indicated that there were two evolutionary branches in the northeastern black bees.【Conclusion】Northeastern black bee has two evolutionary directions, the genetic diversity was high, and because of the high rate of gene flow, the genetic differentiation was moderate among populations. These results could help selective breeding programs, conservation planners design and locate reserves more likely to resist invasion.

Fluorescence Quantitative RT-PCR Assay for Detection of Tembusu Virus

YU Chun-Mei, DIAO You-Xiang, TANG Yi, CUI Jing-Teng, GAO Xu-Hui, ZHANG Ying, JU Xiao-Jun, WU Li-Li

Scientia Agricultura Sinica. 2012, 45(21): 4492-4500. doi:10.3864/j.issn.0578-1752.2012.21.018

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【Objective】The objective of the study is to establish a method for detecting Tembusu virus by SYBR GreenⅠ relative fluorescence quantitative RT-PCR. 【Method】 Special primers based on Tembusu virus NS5 and E gene were designed and a pair of primers of house-keeping gene β-actin was chosen. Then these amplified fragments were cloned into pMD18-T. Using the plasmids NS5-pMD18-T, E-pMD18-T and β-actin-pMD18-T as standard products, a real-time quantitative reverse transcription-polymerase chain reaction ( RT-PCR) was performed to construct the standard curves of NS5, E and β-actin gene and detect the sensitivity, specificity and repeatability. 【Result】 The results showed a precise linear relationship with a correlation coefficient of R²＞0.99. The detection limits was 10 copies of DNA plasmid reaction. The amplification curve showing a single peak could only been detected for Tembusu virus. The variation coefficient was less than 0.5% by within and between the group of repeatability tests. The clinical samples were detected 3 times by this method, and all results were positive.【Conclusion】 The developed real-time PCR assay was highly specific, sensitive, and reproducible and could be an available tool for diagnosis and monitoring of Tembusu virus in duck farms.

Strategies on Integrated Development of Agriculture- Forestry-Grass-Grazing in the Loess Plateau of China

LIU Wei-Jia

Scientia Agricultura Sinica. 2012, 45(21): 4501-4507. doi:10.3864/j.issn.0578-1752.2012.21.019

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Agricultural husbandry in the Loess Platean mainly focus on crop cultivation, but now it is faced with the challenge of sustainable development. The main-accessorial relationship between agriculture and animal husbandry should be adjusted from crop cultivation to grazing. Through coupling development of agriculture-forestry-grass-grazing, reconfigure agriculture and animal husbandry resources. Based on selecting suitable vegetation types (grass, shrub and forest) for different environmental conditions, the advance development of strategy by planting grass and shrub should be carried out to form vertical ecological barrier of grass-shrub-forest and correlative industrial structure gradually. The development of animal husbandry in the Loess Plateau should be eco-friendly type of animal husbandry that has special path and innovation mechanism. Dry-lot feeding or semi-dry-lot feeding mode of husbandry management should be advocated, and with the large scale livestock farmers as the mainstay while cooperatives and breeding areas working separately as ties and platform to develop the subjects of husbandry production and management in the Loess Plateau. Centering on coupling development of agriculture-forestry-grass-grazing, the supporting points such as industry cultivation, policy support, project investment and interest guide should be strengthened, thus form four-wheel driving development pattern of animal husbandry in the Loess Plateau.

Differences in Endowments of Peasant Household, Agricultural Subsidies, and Peasants’ Willingness to Transfer Farmlands —A Micro-Empirical Analysis from a Survey on Peasant Household of 30 Villages in 8 Provinces

HOU Shi-An

Scientia Agricultura Sinica. 2012, 45(21): 4508-4516. doi:10.3864/j.issn.0578-1752.2012.21.020

Abstract ( 825 )

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【Objective】The impacts of endowment differences in farmlands, human resources and property among peasant household and the state’s agricultural subsidy policies on peasants’ willingness to transfer farmlands were studied, and the fundamental factors of the farmlands transfer were analyzed.【Method】On the basis of survey data from 1 585 peasant households of 8 provinces in 2010, this article used Binary quantile regression and bayesian estimation techniques to measure the effect.【Result】 It was found that all kinds of endowment differences and policy variables have an obvious quantile effect on peasant households’ willingness to transfer farmlands.【Conclusion】There is a substitute relationship between farmlands’ social security function and its income effect. It has a better social security function on those who are at disadvantage endowments. Subsidies for growing superior grain and subsidies for agricultural capital goods play a limited role in encouraging peasant household to acquire farmlands. The agricultural subsidy policies should be combined with the career training of rural labors. Small loans play an obvious role in farmlands transfer.

Applicability Screening of Mitigation Technologies and Management Practices of Emissions of Greenhouse Gases from Agriculture Sector

MI Song-Hua, HUANG Zu-Hui

Scientia Agricultura Sinica. 2012, 45(21): 4517-4527. doi:10.3864/j.issn.0578-1752.2012.21.021

Abstract ( 610 )

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【Objective】 Applicability assessment of mitigation technologies and management practices of emissions of greenhouse gases from agriculture. 【Method】 An initial long list of mitigation measures was obtained based on a literature review and expert interview, then the modified delphi method was used to explore expert opinions from interdisciplinary sphere, and a final short list of agricultural mitigation technologies was screened. 【Result】 A final short list of three topics and eighteen measures was drawn up in light of mechanism and measure certainty, technically feasibility, abatement potential, increase or stability in yield, practices easily adopted by farmers, among which cropland nitrous oxide mitigation applicable technologies had an emphasis on nutrient management and farming practices, rice paddy field methane mitigation aimed at water management and cultivar selection and livestock mitigation technologies and management practices focused on forage and manure management.【Conclusion】 Breeding high yield and reduction GHG cultivar should be a core work for agricultural mitigation technology system, meanwhile, supporting subsidy and incentive policies and reinforcing technical training /demonstration should be strengthened in order to inspire the farmers adoption willing.

Research on Path and Key Problem of Coordinated Development of Urbanization, Agricultural Modernization and Industrialization in Central Plains Economic Region

WU Yi-Ping, CHEN Su-Yun, SUN De-Zhong

Scientia Agricultura Sinica. 2012, 45(21): 4528-4533. doi:10.3864/j.issn.0578-1752.2012.21.022

Abstract ( 813 )

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【Objective】This paper was done to study the path and key problem of coordinated development of urbanization, agricultural modernization and industrialization in Central Plains Economic Region.【Method】GM (1, N ) model was used in analysis drive factors and influence factors of coordinated development of urbanization, agricultural modernization and industrialization.【Result】The drive index of urbanization and agricultural modernization is negative, the lowest is the urbanization drive index and the highest is the industrialization drive index.【Conclusion】The path of Central Plains Economic Region development should be led by urbanization and combined with rural community construction. In this process, the following problems need to be solved: Urbanization should be coordinated with industrial agglomeration; land transfer should be moderated and income distribution should be reasonable; urbanization should be pressed gradually to ensure interest in land of farmer; all kinds of funds should be integrated to solve the shortage of funds.

A Dynamic Clustering Method with Missing Data

XIAO Jing, LUO Ru-Jiu, SONG Wen, TANG Zai-Xiang, XU Chen-Wu

Scientia Agricultura Sinica. 2012, 45(21): 4534-4542. doi:10.3864/j.issn.0578-1752.2012.21.023

Abstract ( 666 )

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【Objective】 The aim of the study is to investigate a clustering method for clustering the data with missing values in practice research. 【Method】The paper introduces a maximum likelihood-based dynamic clustering method, which could configure a complete data set through the maximum likelihood estimation for the missing by statistics of the others. The parameters of missing data and different clusters are estimated by the maximum likelihood method implemented via expectation-maximization (EM) algorithm and the objects are classified by the Bayesian posterior probability. 【Result】 The results of simulation studies show that the proposed method not only has fast convergence speed but also accurately cluster the data with missing values. 【Conclusion】The proposed method was further validated by Fisher’s Iris dataset. The result indicated that the proposed method had a significant advantage on clustering accuracy compared to the delete missing data arithmetic and it is similar to complete data clustering algorithm.

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