Scientia Agricultura Sinica

Functional Analysis via Overexpressing Xyloglucan Endotransglycosylase Gene OsXTH11 in Rice

ZHANG Li, NIU Xiang-Li, ZHANG Hui-Ying, LIU Yong-Sheng

Scientia Agricultura Sinica. 2012, 45(16): 3231-3239. doi:10.3864/j.issn.0578-1752.2012.16.001

Abstract ( 1027 )

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【Objective】 The functions of rice XTH gene (OsXTH11) in deveolpment regulation and stress responses were investigated by utilizing transgenic technology. 【Method】 The overexpression vector of OsXTH11 was constructed and introduced into rice cv. Nipponbare by Agrobacterium-mediated transformation. The phenotypes analysis and molecular detection of transgenic plants were performed to verify the function of OsXTH11. 【Result】 Real-time PCR indicated that the expression levels of OsXTH11 in transgenic lines were distinctly increased. The performance of T1 transgenic seedlings revealed that OsXTH11 overexpression lines showed increased growth rates under normal condition, and more responsive to gibberellin (GA) and auxin (IAA). The stem elongation of transgenic plants was significantly faster than that of wild type under dark or deep-water conditions, and exhibited enhanced tolerance to 100 mmol•L-1 NaCl. 【Conclusion】 Phenotypes of transgenic plants overexpressed the xyloglucan endotransglycosylase/hydrolase gene suggested the role of OsXTH11 in rice growth and salt stress.

Analysis of Wheat-Thinopyrum ponticum Translocation Lines with Broad Spectrum Resistance to Stripe Rusts

HAO Wei-Wei, TANG Cai-Guo, LI Bao-Chun, HAO Chen-Yang, ZHANG Xue-Yong

Scientia Agricultura Sinica. 2012, 45(16): 3240-3248. doi:10.3864/j.issn.0578-1752.2012.16.002

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【Objective】To transfer strip rust resistance from Thinopyrum ponticum to common wheat, wide hybrid progenies between them were screened for strip rust resistance.【Method】The progenies were characterized by their disease resistance，agronomic traits，genomic in situ hybridization and quality related protein analysis.【Result】 Eight lines with good resistance or immune to the pathogens at adult stage were obtained. All of them have 42 chromosomes, in which seven lines convey a new translocation, 4DS/4St. In addition, besides this translocation, the GDR3 conveys another terminal translocation, which may be derived from the E-genome chromosomes of Th. ponticum. The GDR3 performed very good resistance in whole life. It was immune to yellow rusts. It was estimated that both of the alien segments convey resistant genes to the stripe rust. The high molecular weight glutenin subunits (HMW-GS) analysis indicated that all of the eight lines convey the good subunits 14+15 at Glu-B1, which should be inherited from Xiaoyan 6. 【Conclusion】 Wild relatives of wheat usually convey genes with broad spectrum resistance to pathogens. This is the genetic base for their surviving in extreme environment, which also provide excellent gene resource for disease resistant breeding in wheat.

A New Strategy for Crop Improvement Through Modification of Phytochrome Signalling Pathways

ZHAN Ke-Hui, LI Zhi-Yong, HOU Pei, XI Yu-Lin, XIAO Yang, MENG Fan-Hua, YANG Jian-Ping

Scientia Agricultura Sinica. 2012, 45(16): 3249-3255. doi:10.3864/j.issn.0578-1752.2012.16.003

Abstract ( 735 )

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In this review, the mechanisms that phytochromes controlling of seedling photomorphogenesis in model plant Arabidopsis were introduced. The present research works on light signaling pathways, mutant discoveries and the relations with yield traits in the grasses were summarized. Overexpression and/or loss-of-function mutants of phytochromes in wheat and maize lead to significant variations in plant height, flowering time and shade avoidance responses. Genetics, biochemical and molecular analyses on these variations related to yield traits will help us develop a new strategy for crop improvement through the modification of light signalling pathways.

Analysis on Grain Filling Characteristics of Indica and Japonica Rice in Rapeseed (Wheat) - Rice Planting Area in Chengdu Basin

LI Xu-Yi, CHI Zhong-Zhi, JIANG Xin-Lu, ZHENG Jia-Guo

Scientia Agricultura Sinica. 2012, 45(16): 3256-3264. doi:10.3864/j.issn.0578-1752.2012.16.004

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【Objective】In order to provide a scientific basis for planting japonica rice in Chengdu basin, the difference in grain filling characteristics and yield formation between indica and japonica rice was studied in rapeseed (wheat) - rice planting area in Chengdu basin.【Method】A field experiment was conducted with indica rice varieties Fuyou 838 and Luyou 578, japonica rice varieties Liaoxing 15 and Liaoxing 19. The grain-filling processes of superior and inferior grains of different varieties were studied with Richards equation. On that basis, the relationship between grain filling characteristics with dry matter accumulation and transformation after heading and yield formation was analyzed in different varieties.【Result】With bigger sink potential and higher seed setting rate, the grain yield of Liaoxing 19 was significantly higher than Liaoxing 15 in japonica rice, even achieved the level of indica rice (Fuyou 838 and Luyou 578).The types of indica and japonica rice initiated their grain filling for superior and inferior grains were synchronous type and asynchronous type, respectively, and the effects of panicle size on grain filling and seed setting were also different between indica and japonica rice. The ability of dry matter transformation after heading for Liaoxing 19 was closer to local indica rice, so it was beneficial to start its grain-filling. Although grains per spike of Liaoxing 19 were significantly higher than Liaoxing 15, the grain filling rate of its inferior grains was not decreased, and the grain filling rate declined slower after reaching peak, grain filling time was also prolonged to some extent, at last the seed setting rate was improved significantly.【Conclusion】 The japonica rice varieties of Liaoxing 19 had more grains per spike, but with the improved dry matter transformation after heading, it still gained higher seed setting rate by better grain-filling. So it has the high yield potentiality for planting in Chengdu basin.

Impacts of Salt Stress on Characteristics of Photosynthesis and Chlorophyll Fluorescence of Sorghum Seedlings

SUN Lu, ZHOU Yu-Fei, LI Feng-Xian, XIAO Mu-Ji, TAO Ye, XU Wen-Juan, HUANG Rui-Dong

Scientia Agricultura Sinica. 2012, 45(16): 3265-3272. doi:10.3864/j.issn.0578-1752.2012.16.005

Abstract ( 810 )

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【Objective】 The impacts of salt stress on the photosynthesis and chlorophyll fluorescence parameters of sorghum seedlings were studied for providing a foundation for sorghum cultivation, breeding and artificial regulation of salt stress.【Method】 Salt tolerant cultivar (Liaoza 15) and salt sensitive cultivar (Longza 11) were incubated in the nutrient solution at humidity of 60%, light/dark of 12 h/12 h, illumination of 134 μmol•m-2•s-1 and 28℃/25℃ of day/night. NaCl was added into the solution at 3-leaf stage and NaCl concentration levels at 0, 50, 100, 150 and 200 mmol•L-1, respectively. The response of sorghum seedlings to NaCl stress was assessed by measuring the photosynthesis and chlorophyll fluorescence parameters of seedlings. 【Result】 Low NaCl concentration (50 mmol•L-1) increased the chlorophyll content, and high NaCl concentration (100-200 mmol•L-1) reduced the content substantially. Salt stress reduced Pn, Gs, Tr, Fm, Fv/Fo, Fv/Fm, Fv′/Fm′ and qP, and increased Fo and NPQ. Low NaCl concentration (50 mmol•L-1) reduced Ci, and high NaCl concentration (100-200 mmol•L-1) increased it. The adverse impact of salt stress on Liaoza 15 was less than on Longza 11. 【Conclusion】 The small reduction in net photosynthesis rate caused by 50 mmol•L-1 NaCl stress was considered to be a result of non-stomatal restriction; but increased stomatal restriction with increased NaCl concentration resulted in more severe reductions in photosynthesis. Under salt stress, salt tolerant cultivar could protect the photosynthetic organs more effectively than salt sentitive cultivar and thus improve the production of sorghum in salt affected areas.

Genetic Analysis and Molecular Mapping of Leaf Rust Resistance Genes in Two Chinese Wheat Lines

ZHOU Yue, WU Yu, LI Xing, LI Zai-Feng, LIU Da-Qun

Scientia Agricultura Sinica. 2012, 45(16): 3273-2380. doi:10.3864/j.issn.0578-1752.2012.16.006

Abstract ( 824 )

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【Objective】 The objective of this study is to identify the leaf rust resistance genes of two Chinese wheat (Triticum aestivum L.) lines SW8588 and Mianyang 351-15 from Sichuan province, and to provide a basis for breeding wheat cultivars with durable resistance. 【Method】 Mianyang 351-15, SW8588 and 30 differential lines with known leaf rust resistance genes were inoculated with 15 pathotypes of Puccinia triticina for postulating leaf rust resistance genes in the two lines. The two F1 and F2 populations from crosses of Mianyang 351-15/Zhengzhou 5389 and SW8588/Zhengzhou 5389, were inoculated with pathotype FHTT in the greenhouse. The infection types were scored 15 days after inoculation and molecular markers were also used for mapping the resistance genes.【Result】SW8588 did not contain the known leaf rust resistance gene Lr1 and Mianyang 351-15 maybe contain Lr1 and other unknown genes. When inoculating with pathotype FHTT, only one resistance gene was detected in each of the two resistance lines. Using SSR makers, the resistance gene in Mianyang 351-15 was mapped on 5DL and linked to SSR markers barc144 and wmc765 with genetic distances of 8.9 and 20.8 cM, respectively. The STS marker WR003 for Lr1 was co-segregated with this resistance gene in Mianyang 351-15. Based on these results, it was concluded that Mianyang 351-15 contained Lr1 conferring the resistance to FHTT. Using genetic analysis and molecular markers it could be found that SW8588 carried a single dominant resistance gene, temporarily designated LrSW85, co-segregating with STS marker WR003, which may be an allele of Lr1 or was closely linked to Lr1.【Conclusion】Based on gene postulation, genetic analysis and molecular mapping it can be concluded that Mianyang 351-15 contained the leaf rust resistance gene Lr1 and SW8588 contained the leaf rust resistance gene LrSW85 which may be an allele of Lr1 or closely linked to Lr1.

Transcription Activity of a Transcription Factor StSte12 from Setosphaeria turcica and Function Analysis Through Yeast Complementation

WU Min, GU Shou-Qin, LI Po, WANG Mei-Juan, ZHANG Chang-Zhi, FAN Yong-Shan, HAN Jian-Min, DONG Jin-Gao

Scientia Agricultura Sinica. 2012, 45(16): 3281-3287. doi:10.3864/j.issn.0578-1752.2012.16.007

Abstract ( 559 )

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【Objective】 The objective of this study is to identify the structure characteristics, transcription activity and function of StSte12 from Setosphaeria turcica. 【Method】Conservative domain prediction and phylogenetic tree analysis were used to predict the possible function of StSte12 through bioinformatics analysis. The transcription activity of StSTE12 was tested by β-galactosidase activity method. Through characterization of growth of the StSTE12 complement transformant, in which StSTE12 was introduced into ScSTE12 null mutant of Saccharomyces ceresivisiae, the function of StSTE12 was preliminarily identified.【Result】StSte12 contained STE homeodomain and ZnF_C2H2¬ structure, which was the characteristics of Ste12-like transcription factor. Sequence analysis revealed that StSTE12 shared highly homology with other STE12-like genes of plant pathogens. StSte12 had transcription activation in vitro and could restore the function of ste12Δ of S. ceresivisiae, which regulated invasion growth of the yeast cell. 【Conclusion】Transcription factor StSTE12, a STE12-like gene from S. turcica, has a transcription activity and plays an important role in regulating the invasion growth of the cell on YPD medium.

Construction of Recombinant BmNPV Infecting Ecotropis oblique

GUO Rui, LIU Wei-Xing, PAN Zhong-Hua, XUE Ren-Yu, CAO Guang-Li, ZHU Yue-Xiong, GONG Cheng-Liang

Scientia Agricultura Sinica. 2012, 45(16): 3288-3296. doi:10.3864/j.issn.0578-1752.2012.16.008

Abstract ( 730 )

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【Objective】The objective of this study is to construct recombinant BmNPV infecting both silkworm and Ecotropis oblique, and to solve the reproduction difficulty of EoNPV because of the breeding complexity.【Method】Recombinant BmNPV containing the polh expression box of BmNPV and the hel expression box of EoNPV were constructed by Bac-to-Bac system. 【Result】rBmNPV amplified from silkworm or BmN cell could infect E. oblique, which indicated the host domain of rBmNPV had been broaden to E. oblique. 【Conclusion】rBmNPV which could infect E. oblique was successfully constructed, thus laied a foundation of producing recombinant virus pesticide using silkworm as host.

Mining the Accumulated Nitrate from Deep Soil Layers by Rotation with Different Crops

ZHANG Yong-Li, JU Xiao-Tang

Scientia Agricultura Sinica. 2012, 45(16): 3297-3309. doi:10.3864/j.issn.0578-1752.2012.16.009

Abstract ( 651 )

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【Objective】This study aimed to use plants in cropping systems for bioremediation, depleting nitrate nitrogen in deep soil profile to control nitrate leaching caused by excessive nitrogen fertilizer application.【Method】Different treatments were designed in a plot experiment, including conventional wheat-maize rotation system, fallow-summer maize system, winter wheat-fallow system, rye-amaranth rotation system, rye-sorghum hybrid sudan grass rotation system , rye-sweet sorghum rotation system, alfalfa continuous cropping and alfalfa+fescue inlercropping. The effects of each cropping system on the decrease of accumulation and leaching of soil nitrate nitrogen was analyzed.【Result】The results show that alfalfa, sorghum hybrid sudan grass and rye had the highest percentage of roots in 1-2 m soil profile. Annual plant N uptake under rye-amaranth, rye-sorghum hybrid sudan grass and rye-sweet sorghum treatments were the highest, about 300-390 kgN•hm-2. Fallow in summer season increased nitrate leaching from 0-1 m soil profile. After the first year’s remediation, all the five treatments significantly reduced nitrate accumulation in 0-1 m and 0-2 m soil profile, decresed 124.3 kgN•hm-2 and 81.2 kgN•hm-2 respectively. Sorghum hybrid sudan grass, sweet sorghum and amaranth were more effective in decreasing nitrate nitrogen accumulation in deep soil layers. Nitrate concentration in soil solution under rye-sweet sorghum, rye-sorghum hybrid sudan grass and rye-amaranth systems was in the lowest level, averaged only 8.6 mg•L-1.【Conclusion】According to the first year’s results, rye-sorghum was the best cropping system to decrease the accumulated nitrate in deep soil layers and further control nitrate pollution.

Effect of Application of Silicon Amendment on Cd Over-Standard Soils of Vegetable Fields

WANG Yan-Hong, AI Shao-Ying, TANG Ming-Deng, LI Meng-Jun, YAO Jian-Wu, LUO Ying-Jian

Scientia Agricultura Sinica. 2012, 45(16): 3310-3317. doi:10.3864/j.issn.0578-1752.2012.16.010

Abstract ( 849 )

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【Objective】The purpose of this study was to explore the effects of application of different amounts of silicon amendment on Cd uptake by leafy vegetables and soil available Cd concentration in cadmium over-standard vegetable field and to provide reference for proper use of silicon amendment to reduce leafy vegetable Cd concentration.【Method】Two field experiments were carried out to investigate Cd bioavailibity in soils and Cd accumulation by leafy vegetables under four levels of silicon amendment application in Cd over-standard fields in the suburbs of Guangzhou in 2009.【Result】The results showed that compared with the control silicon amendment insignificantly increased the output of flowering Chinese cabbage and lettuce grown in Cd over-standard soils, while shoot Si concentrations of flowering Chinese cabbage and lettuce significantly increased by 1.00- and 2.40-fold in the silicon amendment at 60 kg?667m-2 treatment, respectively, and their root of Si concentrations increased by 0.95- and 2.37-fold, respectively. In comparison with the control, the silicon amendment at 60 kg?667m-2 significantly reduced shoot and root Cd concentrations of flowering Chinese cabbage and lettuce, with shoot Cd concentration reducted by 25% and 32% and root by 17% and 15% respectively. Compared with the control, silicon amendment decreased soil DTPA-Cd concentrations significantly, and insignificantly affected soil pH of No. 1 vegetable field, but significantly increased soil pH of No. 2 with no significant differences between application levels.【Conclusion】At the application level of 60 kg?667m-2, silicon amendment could significantly reduce plant Cd content, and might be used as an effective soil amendment to inhibit Cd uptake by vegetables in Cd over-standard soil.

Diversity and Genetic Structure of Garlic (Allium sativum L.) Germplasm Resource in China

WANG Hai-Ping, Philipp W.Simon, LI Xi-Xiang, CHENG Jia-Qi, SHEN Di, SONG Jiang-Ping, QIU Yang, ZHANG Xiao-Hui

Scientia Agricultura Sinica. 2012, 45(16): 3318-3329. doi:10.3864/j.issn.0578-1752.2012.16.011

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【Objective】 The objective of the experiment is to understand the diversity and genetic structure of garlic germplasm resources in China. 【Method】 Two-hundred and twelve accessions of garlic germplasm preserved in China were detected and analyzed based on AFLP, SSR and InDel. Neighbor-joined clustering was analyzed by software Mega, and the population genetic structure was studied by software Structure 2.1. The effect of genetic structure on association analysis between the molecular markers and 21 quantitative traits including allicin content was evaluated by general linear model (GLM) by software SSPS13.0 .【Result】Totally, 502 allels were amplified by AFLP, SSR and InDel primers, and 492 of them were polymorphic among 212 accessions of garlic. All accessions were divided into 5 groups by both structure analysis and neighbor-joining clustering. However, the Shannon’s index of each group assumed by genetic structure analysis was smaller than that assumed by neighbor-joining clustering, which indicated that the genetic structure analysis could interpret genetic relationship among the individual accessions in more details. Most of traits including allicin content were slightly affected by population structure, which indicated that the germplasm in this study was acceptable to be the populations for association mapping.【Conclusion】The garlic germplasm resources in China possess a great diversity. The population structure of the garlic germplasm resources slightly affect the quantitative traits including the allicin content, which means that the population of the garlic germplasm is a potential candidate natural population for association analysis between the molecular markers and the quantitative traits.

Expression of Genes Related to Starch Synthesis During Rhizome Swelling of Lotus Root (Nelumbo nucifera Gaertn)

CHENG Li-Bao, LI Shu-Yan, LI Yan, YIN Jing-Jing, CHEN Xue-Hao, LI Liang-Jun

Scientia Agricultura Sinica. 2012, 45(16): 3330-3336. doi:10.3864/j.issn.0578-1752.2012.16.012

Abstract ( 755 )

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【Objective】 Critical genes relevant to starch synthesis were explored by studying gene expression during swelling of lotus rhizome. 【Method】 In this study, the changes of starch content and starch granule characteristics were studied during (cv. Mei Ren Hong) swelling period of lotus rhizome. Four important related genes were identified based on transcriptome analysis, digital gene expression profiling and qRT-PCR method. 【Result】 The result showed that content of the starch was improved more rapidly, and the starch granule was more enlarged at swelling stage than that at initial swelling stage. A total of 34 related genes to starch metabolism were identified during this process. However, only four genes including LrGBSS，LrSBEI，LrSBEII and LrSBEIII were obviously changed their transcriptional level at the swelling stage compared with initial swelling period. The result of qRT-PCR was further proved that LrGBSS，LrSBEI，LrSBEII and LrSBEIII showed a high correlation with starch synthesis according to their expression characteristics during rhizome development. 【Conclusion】The expression of LrGBSS，LrSBEI，LrSBEII and LrSBEIII is a critical factor to decide the starch content of lotus rhizome.

Method of Making Pear Fruit-Derived Nutrient Solution and Its Effect on Photosynthetic Characteristics and Fruit Quality

LUO Rui, ZHANG Jie, LIU Ming-Juan, JI Qian-Long, YAO Yun-Cong

Scientia Agricultura Sinica. 2012, 45(16): 3337-3345. doi:10.3864/j.issn.0578-1752.2012.16.013

Abstract ( 987 )

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【Objective】The purpose of this study is to explore the optimal method of making pear fruit-derived nutrient solution and its effect on photosynthetic characteristics and fruit quality.【Method】In the present study, the dropping and thinning fruits from pear tree, Pyrus pyrifolia(Burm.f.).cv. Nakai, were used as materials to make the nutrient solutions and different formula including content of sugar and water, disintegrating status of fruit, anaerobic or aerobic condition and fermentation days were designed to study their effects on the nutrition components in the solutions. Furthermore, in order to analyze the mineral element content, growth status and the photosynthetic characteristics of pear trees, a pear fruit-derived nutrient solution was made based on the optimal formula and sprayed with different concentrations of the pear fruit-derived nutrient solution (Tr1 for 500 times, Tr2 for 200 times, and Tr3 for 0 times as CK) on two-year-old pear trees and fifteen-year-old mature trees.【Result】The results indicated that young pear fruit, ferments, brown sugar and water according to the mass ratio of 50:1:10:35 in the fermentation conditions of fruit homogenate, aerobic conditions at room temperature of 25-28℃ for 30 to 45 days were the most suitable formula to make fruit-derived nutrient solution, and spray with pear fruit-derived nutrient solution significantly increased leaf thickness, the specific leaf weight, the content of chlorophyll, and the content of mineral element, such as N, P, K, Ca, Fe, Mn and so on. In addition, spray with pear fruit-derived nutrient solution not only enhanced the water use efficiency through increased the net photosynthetic rate and decreased the transpiration rate, but also significantly increased the average single fruit weight, fruit-based index, fruit hardness, soluble solids, titratable acid, sugar acid ratio and Vc content.【Conclusion】In conclusion, spraying on the surface of leaves significantly promoted the growth of pear trees, increased the content of mineral element in leaves, enhanced the photosynthetic performance of leaves, further then improved the quality of fruits, especially the 200 times nutrient solution. And the product made by the optimal formula had higher content of mineral elements, degree of decomposition and content of sugar than others.

Degradation of Plastid Pigment and Its Relationship with Volatile Catabolite Content in Cured Leaves of Different Genotypes of Flue-Cured Tobacco

SHI Hong-Zhi, GU Shao-Long, DUAN Wei-Dong, WANG Jian-An, QIAN Hua, LIU Qing-Hua, SU Fei, LIU Guo-Shun

Scientia Agricultura Sinica. 2012, 45(16): 3346-3356. doi:10.3864/j.issn.0578-1752.2012.16.014

Abstract ( 818 )

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【Objective】 The research was conducted to investigate the changes in plastid pigment contents during middle leaf mature and curing, and the relationship of pigment degradation amount and rate to volatile pigment catabolite content in cured leaves. 【Method】HPLC method was used to measure the pigment and volatile aroma components in 9 genotypes of flue-cured tobacco grown in central Henan Province. 【Result】The results showed that the plastid pigment contents decreased during leaf maturing and curing, with the total degradation amount and rate of chlorophyll being greater than that of carotenoids, and that of chlorophyll a being significantly greater than that of chlorophyll b, resulting in relatively higher amount of residue of chlorophyll b in cured leaves. The degradation rate of chlorophyll during leaf maturing was greater than that during curing. Among the carotenoids, lutein had the highest content and greatest degradation amount, and that during leaf maturing was greater than that during curing. The degradation rate of neoxanthin during maturing was also greater that during curing, while it was the contrary for β-carotene and violaxanthin. Different genotypes had different pigment degradation amounts and had significantly positive correlations with the contents of volatile degraded aroma compounds in cured leaves, while there was no or negative correlations with the contents of pigment residue in cured leaves. Total amount of pigment-degraded aroma compounds and the contents of most important individual aroma components had greater positive correlations with degradation amount during maturing than that during curing. 【Conclusion】 The degradation amount of chlorophyll, luten and neoxanthin were greater than that during curing. Total amount of pigment degraded aroma compounds had a close relationship with the pigment degradation amount, especially that during maturing. Thus the increase of maturity of tobacco was pivotal in promoting the formation of aroma components in tobacco leaves.

Resistance of Harvested Fruits and Vegetables to Diseases Induced by ASM and Its Mechanism

GE Yong-Hong, BI Yang, LI Yong-Cai, WANG Yi

Scientia Agricultura Sinica. 2012, 45(16): 3357-3362. doi:10.3864/j.issn.0578-1752.2012.16.015

Abstract ( 1007 )

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Acibenzolar-S-methyl (ASM) is a functional analogue of salicylic acid and an artificial synthetic chemical elicitor. ASM has been shown to induce resistance in a wide range of fruit and vegetables. Spraying with ASM before harvest or postharvest dipping treatment decreased postharvest diseases of pear, peach, grape, strawberry, muskmelon, mandarin, banana, loquat, tomato and potato. The resistance induced by ASM is related to various defense responses, including induction of reactive oxygen species accumulation, activation of phenylpropanoid pathway and production of pathogenesis-related proteins. Range of fruit and vegetables of ASM application, the efficacy of ASM and mechanisms involved in induced resistance with ASM are still need to be investigated in future researches.

Major Bio-Factors Affecting Beef Color Stability

CHEN Jing-Yi, NIU Li, HUANG Ming, ZHOU Guang-Hong

Scientia Agricultura Sinica. 2012, 45(16): 3363-3372. doi:10.3864/j.issn.0578-1752.2012.16.016

Abstract ( 836 )

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【Objective】 The experiment was designed to investigate the major bio-factors affecting color stability from different bovine muscles.【Method】 Three representative bovine muscles– M. longissimus lumborum (LL), M. semimembranosus (SM), and M. psoas major (PM) were obtained from 8 beef carcasses. Instrumental color, pigment content, pH value, MDA content, NADH concentration, LDH and metmyoglobin reductase activity were measured every two days over a period of 7d retail display at 0-4℃.【Result】The order of color stability of the three muscles was: LL＞SM＞PM. The a* value, NADH concentration, LDH and metmyoglobin reductase activity was the highest in LL and the MDA content was the least. While the a* value, NADH concentration, LDH and metmyoglobin reductase activity of PM was the lowest and MDA content was the highest. 【Conclusion】NADH concentration, LDH and metmyoglobin reductase activity in beef muscle have a significant correlation with color stability（P＜0.05）.

Determination of Acrylamide in Kitchen Waste Using Gas Chromatography-Tandem Mass Spectrometry

CHENG Jie, SU Xiao-鸥, ZHANG Wei, WANG Rui-Guo, WANG Pei-Long, LI Yang

Scientia Agricultura Sinica. 2012, 45(16): 3373-3381. doi:10.3864/j.issn.0578-1752.2012.16.017

Abstract ( 630 )

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【Objective】The objective of this study is to develop an analysis methods of the acrylamide in kitchen waste using gas chromatography-tandem mass spectrometry.【Method】The use of warm deionized water extraction and C18 SPE clean-up or the formic acid extraction and Carrez reagent/MCX clean-up as the sorbented appeared to be a very effective sample preparation method. After derivative reaction, gas chromatography-tandem mass spectrometry was used to detect the sample in the mode of SRM, and the external standard method was used for quantitative analysis. The qualitative ion is m/z 152＞135 and 152＞107 and the quantitative ion is m/z 152＞135. The pressure of gas collision is 15V.【Result】The developed method presented a good linearity from 3 ng•L-1 to 30 ng•L-1 with the correlation coefficient of 0.9984. The RSD of the method was 4.3% and of the quantitative limit was 1.0 ng•g-1. The rate of recovery was 95.3%-108.1%.【Conclusion】The developed method is a very effective method for detecting the acrylamide in kitchen waste.

Effects of Feeding Purified Zearalenone Contaminated Diets with or Without Modified Montmorillonite on Immunological Measurements of Postweaning Piglets

JIANG Shu-Zhen, YANG Wei-Ren, YANG Zai-Bin, WANG Shu-Jing, LIU Fa-Xiao, ChiF

Scientia Agricultura Sinica. 2012, 45(16): 3382-3390. doi:10.3864/j.issn.0578-1752.2012.16.018

Abstract ( 693 )

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【Objective】The aims of the present study were to investigate the toxicity of zearalenone (ZEA) on haematology, lymphocyte subset in peripheral blood, serum immunoglobulin and swine plague antibody titer production of piglets, and the efficacy of Calibrin-Z in preventing ZEA-induced adverse effects was also evaluated. 【Method】The piglets were randomly divided into 6 treatment groups and fed individually in a metabolic cage for 22 days after 7-d adaptation. Piglets in group 1 were fed with basal diet only (Control). Piglets in group 2 were fed with basal diet+1.0 g•kg-1 Calibrin-Z, that in group 3 were fed with basal diet+1.0 mg•kg-1 ZEA, that in group 4 were fed with basal diet + 1.0 mg•kg-1 ZEA + 1.0 g•kg-1 Calibrin-Z, that in group 5 were fed with basal diet + 1.0 mg•kg-1 ZEA + 2.0 g•kg-1 Calibrin-Z, and that in group 6 were fed with basal diet + 1.0 mg•kg-1 ZEA + 4.0 g•kg-1 Calibrin-Z. 【Result】Piglets fed ZEA contaminated diet without Calibrin-Z showed significant decreases in platelet, lymphocyte rato, hemoglobin, CD4+ and CD8+ in peripheral blood, serum IgG and swine plague antibody titer production at 21-d when compared with the control (P＜0.05). The evident improved effect was observed when addition of 2.0 to 4.0 g Calibrin-Z/kg diet in 1.0 mg•kg-1 ZEA contaminated diet. Platelet, lymphocyte rato, hemoglobin, and serum IgG (P＜0.05) showed a linear higher effect with increased Calibrin-Z (P＜0.05).【Conclusion】The above findings indicated that ZEA at a level of 1.0 mg•kg-1 was sufficient to induce immunotoxicity of piglets. Addition of 2.0 to 4.0 g Calibrin-Z/kg diet in 1.0 mg•kg-1 ZEA contaminated diet can effectively protect against the detrimental effects of the ZEA feeding. These results may have implications for humans health and animals production consuming ZEA-contaminated food or feed.

Influence of High Temperature on Oxidative Damage and Follicle Development of Laying Hens

YANG Xiao-Lan, ZHANG Min-Hong, FENG Jing-Hai, DI Li-Li, JIANG Li-Wen

Scientia Agricultura Sinica. 2012, 45(16): 3391-3398. doi:10.3864/j.issn.0578-1752.2012.16.019

Abstract ( 939 )

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【Objective】 Two experiments were conducted to assess the relationship between follicle development of heat stressed laying hens and oxidative stress induced by high temperature, and then to reveal the mechanism of lower egg production of heat stressed laying hens.【Method】In experiment 1, 180 Hy-line Brown hens, 27 wk of age, were randomly divided into three groups and housed into three controlled environment chambers. Three groups were respectively normal thermal group (NT group: 28℃, basal diet), daily cyclic high temperature group (HT group: 28-35-28℃, basal diet) and daily cyclic high temperature and supplemented with Vitamin E (VE) group (HT+VE group: 28-35-28℃, basal diet supplemented with 250 IU•kg-1 VE). In experiment two, granulosa cells dissociated from F1 and F2 follicles were incubated under either 39℃ or 44℃. 【Result】The results of experiment 1 showed that daily cyclic high temperature significantly inhibited follicle development of laying hens, which mainly manifested as decreasing in egg production (P＜0.05), ovary weight (P＜0.05) and large follicle number (P＜0.05) as well as inhibition in P450 cholesterol side-chain cleavage enzyme (P450scc ) mRNA expression (P＜0.05) and progesterone (P4) production (P＜0.05) in granulosa cell. The results also showed that daily cyclic high temperature induced hens oxidative stress, which manifested as higher malondialdehyde (MDA) concentration (P＜0.05) in plasma, liver tissue and egg yolk. When exposed to cyclic high temperature, 250 IU•kg-1 VE supplementation decreased MDA concentration significantly in plasma (P＜0.05), liver tissue (P＜0.05), egg yolk (P＜0.05), and alleviated (P＜0.05) inhibition in follicle development of laying hens. The results of experiment 2 showed that high temperature increased reactive oxygen species (ROS) level (P＜0.05) in granulosa cells and decreased proliferation of granulosa cells (P＜0.05). VE supplemented into medium alleviated the negative effects of chronic heat stress, manifested as decreasing in ROS level (P＜0.05) and increasing in proliferation of granulosa cells (P＜0.05). 【Conclusion】 The results demonstrated that heat stress increased ROS level in granulosa cell and induced hens (liver, plasma and egg yolk) oxidative stress, in addition, heat stress inhibited P4 production, proliferation of granulosa cells and follicle development of laying hen, while VE depressed oxidative damage induced by heat stress, promoted follicle development of heat stressed laying hens. All the results suggest that the mechanism of poor follicle development of heat stressed hens involves in oxidative stress induced by high temperature.

Effect of Plant Essential Oil on the Distribution of Urea-Nitrogen in Rumen Contents

ZHAO Sheng-Guo, WANG Jia-Qi, BU Deng-Pan, ZHOU Ling-Yun, SUN Peng

Scientia Agricultura Sinica. 2012, 45(16): 3399-3405. doi:10.3864/j.issn.0578-1752.2012.16.020

Abstract ( 794 )

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【Objective】 The objective of the experiment is to reveal the distribution of urea-nitrogen in rumen contents and the effect of essential oil on the distribution. 【Method】 Garlic oil, tea tree oil and eucalyptus oils were added into serum bottles containing rumen fluid and 15N-urea. The abundance of 15N and rumen fermentation were detected after incubation. The bacterial population was studied by DGGE.【Result】 Urea nitrogen in rumen fluid decreased and that in the microbes associated with rumen solid and liquid phases increased following the incubation time, and then had the similar level at 24 h. Compared to the control, urea nitrogen abundance in the microbes associated with rumen solid and liquid phases treated with garlic oil increased by 22.60% and 18.75%, pH and ammonia nitrogen decreased by 3.02% and 17.80%, respectively. The proportion of isobutyric acid and acetic acid: propionic acid increased at 24 h. The pH in rumen fluid treated with tea tree and eucalyptus oils decreased (P＜0.05), and the proportion of isobutyric acid and acetic acid:propionic acid increased. Three different essential oils affected bacterial population according to the results by DGGE.【Conclusion】 Urea-nitrogen was transferred from rumen fluid to microbes, and garlic oil inhibited the transfer, changed microbial population and decreased ammonia production.

Study on Cytotoxicity of Outer Membrane Protein BP26 of Brucella

CHEN Rui-Hua, ZHANG Hui, TANG Li-Yan, MENG Ru, ZHANG Yu, WANG Zhen, LI Zhi-Qiang, ZHANG Jun-Bo, CHEN Chuang-Fu

Scientia Agricultura Sinica. 2012, 45(16): 3406-3413. doi:10.3864/j.issn.0578-1752.2012.16.021

Abstract ( 722 )

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【Objective】 Outer membrane protein BP26 of Brucella is an important Brucella virulence factor. In order to reveal the biological functions of Brucella bp26 gene, HPT-8 cells were infected by bp26 gene deletion strains and treated by BP26 protein respectively. 【Method】 The BP26 protein was purified by Ni affinity chromatography and bp26 gene deletion strains was constructed by overlap technology. After the infection of bp26 gene deletion strains and the treatment of BP26 protein, HPT-8 cells were observed by electron microscope to examine the cells morphology. ELISA was used to detect cytokines in the supernatant.【Results】bp26 gene was cloned from Brucella vaccine strain M5-90 and successfully expressed in E. coli BL21(DE3).Purified protein was proved correct by SDS-PAGE, and the immunogenicity of the obtained BP26 was confirmed by Western-blot. The upper and lower arms of bp26 gene were inserted into the suicide vector PGEM-7zf, transferred to Brucella vaccine strains M5-90 and the bp26 deletion strains with genetic stability were selected successfully. The treatment of BP26 protein changed the cells morphology and significantly decreased the adhesion ability to the wall. Compared with the PBS group, the release level of IL-6,TNF-α and LDH significantly increased（P＜0.01）, but the release level of IL-10 significantly decreased (P＜0.05). After the infection of bp26 gene deletion strains, the release level of IL-6, TNF-α were higher than M5-90 group. The release level of LDH and IL-10 were significantly lower than M5-90 group（P＜0.05）.【Conclusion】 BP26 protein and Brucella M5-90Δbp26 deletion strains were successfully obtained. BP26 protein could lead to inflammatory response and had some toxic effects on the HPT-8 cells. The bp26 gene plays an important role in Brucella infection of HPT-8 cells.

Construction of miR-34c Lentiviral Expression Vector and Its Infection in Dairy Goat Germ Line Stem Cells

LIU Chao, YU Meng, ZHU Hai-Jing, LI Ming-Zhao, HUA Jin-Lian

Scientia Agricultura Sinica. 2012, 45(16): 3414-3421. doi:10.3864/j.issn.0578-1752.2012.16.022

Abstract ( 644 )

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【Objective】In order to over-express mouse miR-34c economically and efficiently, mouse miR-34c lentiviral expression vector was constructed using two different methods.【Methods】The pri-miR-34c was amplified and inserted into downstream of CMV and U6 promotors in pLL3.7, respectively. The recombinant lentiviral vector together with lentivirus package plasmid mixtures were transfected into 293T cells to package virus. The titres of the viruses were examined and then the viruses were infected with 293T cells and dairy goat male germ line stem cells and 293T cells. The transfection efficiency was measured by the percentage of GFP expression cells, and the expression level of miR-34c was determined by qPCR.【Results】The recombinant pLL3.7-CMV-34c vector was successfully constructed, confirmed by endonuclease digestion analysis and DNA sequencing. The expression level of miR-34c in cells infected with the packaged pseudoviriius was distinctively increased. 【Conclusion】Using both CMV and U6 promotor can over-express miR-34c vector. And the virus packaged with recombined vector using U6 promoter can infect dairy goat male germ line stem cells more efficiently.

Epidemiological Survey and Identification of Theileria Parasite Infection for Small Ruminants in Some Parts of China

LI You-Quan, PENG Yu-率, LIU Zhi-Jie, GUAN Gui-Quan, YANG Ji-Fei, CHEN Ze, NIU Qing-Li, LUO Jian-Xun, YIN Hong

Scientia Agricultura Sinica. 2012, 45(16): 3422-3429. doi:10.3864/j.issn.0578-1752.2012.16.023

Abstract ( 684 )

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【Objective】The objective of this experiment is to investigate the current epidemiological status of ovine and caprine theileriosis and identified its pathogens in China. 【Method】The genomic DNA of the blood samples of sheep and goats and ticks were amplified using specific primers to T. uilenbergi, T. luwenshuni and T. ovis, respectively. The 18S rRNA gene sequences from the positive samples were obtained and used for phylogenetical analysis.【Result】On the basis of the PCR results, wide spreading of the theileriosis and significant difference in terms of pathogen species and prevalence in the investigated sites were observed. Co-infection of T. uilenbergi and T. luwenshuni was found only in Gansu Provinces. T. ovis infection was found in Kashi, Xinjiang, but T. luwenshuni infection was detected in Hubei Province but no clinical cases were found. None Theileria infection in Yunnan Province was found. 【Conclusion】The prevalence and infection situation of ovine and caprine theileriosis varied markedly in the studied sites. The data will be useful for guiding the comprehensive prevention and control of ovine and caprine theileriosis in these regions.

Isolation and Functional Analysis of the Low-Temperature-Responsive Gene MtbHLH-1 in Medicago truncatula

LI Yu-Wei, LIAN Rui-Li, WANG Xin-Min

Scientia Agricultura Sinica. 2012, 45(16): 3430-3436. doi:10.3864/j.issn.0578-1752.2012.16.024

Abstract ( 755 )

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【Objective】This study is aimed to characterize the low-temperature-responsive gene MtbHLH-1 in M. truncatula by clone and analyse its sequence, and study the function of MtbHLH-1 in low temperature response. 【Method】The full-length cDNA sequence of MtbHLH-1 was isolated by RT-PCR. The obtained cDNA sequence and the deduced amino acid sequence were analysed. Semi-quantitative RT-PCR was used to assess the expression of MtbHLH-1 in response to low temperature (4℃) stress. 【Result】The sequencing result showed that the cloned cDNA (designated as MtbHLH-1, GenBank Accession No. JN800447) was 778 bp, contains an open reading frame (ORF) of 741 bp, and encoding a protein with 246 amino acids. BLASTp search revealed 45% to 73% similarities between the MtbHLH-1 protein and those reported bHLHs from other species, such as Glycine max, Arabidopsis thaliana, Nicotiana tabacum, Triticum aestivum and Zea mays. The qRT-PCR result showed that MtbHLH-1 was expressed upregulatedly in response to low temperature stress.【Conclusion】The cold stress response transcription factor MtbHLH-1 was cloned from M. truncatula and the semi-quantitative analysis of the gene MtbHLH-1 under low temperature demonstrate that the MtbHLH-1 gene probably improved low temperature stress tolerant capacities.

Effects of Density on Maize Yield and Yield Components

WANG Kai, WANG Ke-Ru, WANG Yong-Hong, ZHAO Jian, ZHAO Ru-Lang, WANG Xi-Mei, LI Jian, LIANG Ming-Xi, LI Shao-Kun

Scientia Agricultura Sinica. 2012, 45(16): 3437-3445. doi:10.3864/j.issn.0578-1752.2012.16.025

Abstract ( 755 )

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【Objective】Field experiments were conducted to study the influence of density on maize yield (＞15 000 kg?hm-2) and yield components in order to exploring the formation mechanism of high-yield so as to provide a basis for stable high-yield maize.【Method】 The experiments were carried out in high-yield maize areas of Xinjiang and Ningxia regions. Zhengdan 958 (ZD958) was planted at different densities from 15 000 plants/hm2 to 180 000 plants/hm2 by making 15 000 plants/hm2 as a density gradient with fully satisfy fertilizer and water demand. High-yield cultivation was practiced, and then the yield and yield component, characteristics were analyzed based on the high-yield (＞15 000 kg?hm-2) .【Result】The 68 different density level plots were all at mutil-sites across two years, the highest and lowest yields were between 7 675.5 kg?hm-2 and 20 503.5 kg?hm-2, respectively. Yields of the 47 plots achieved the high yield of above 15 000 kg?hm-2. Plots with at the highest and lowest densities (52 500 plants/hm2 and 162 800 plants?hm-2) achieved the high yield of 15 000 plants/hm2. The highest and lowest number of ears was 66 600 ears/hm2 and 138 400 ears/hm2, respectively. The highest and lowest grains per ear were 365 and 657, respectively. The highest and lowest 1 000-seed-weight was 273 g and 404 g, respectively.【Conclusion】The parabolic relationship between density and yield was found, and the highest yield appeared at the density of 105 000 plants/hm2. With the increase of production, the optimal range of planting density, ear number, grains per ear and 1000-seed-weight were narrowed. With the increase of planting density, grains per ear and 1 000-seed-weight declined; ears number and grains/hm2 increased, but grains/hm2 increased slightly when density reached a certain level.

Regional Pattern Changes of Rice Production in Thirty Years and Its Influencing Factors in Jiangsu Province

ER Jun, ZHANG Hong-Cheng, LU Jian-Fei

Scientia Agricultura Sinica. 2012, 45(16): 3446-3452. doi:10.3864/j.issn.0578-1752.2012.16.026

Abstract ( 671 )

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【Objective】In order to provide a basis of scientific decision-making for sustainable development of rice production in Jiangsu Province,the regional pattern changes of rice production from 1980 to 2009 in Jiangsu Province and its influencing factors were analyzed for exploring the influence of acceleration of industrialization and urbanization on regional pattern changes of rice production in Jiangsu Province. 【Method】Based on the statistical data from 1980 to 2009, regional pattern changes of rice production in Jiangsu Province and its influencing factors were discussed with the methods of mathematical statistics and regional difference in recent thirty years. 【Result】In the last thirty years, great changes had taken place in rice planting area of different regions in Jiangsu Province, with the characteristics of quick decrease in South Jiangsu area, slight decrease in Central Jiangsu area, and quick increase in North Jiangsu area. Rice yield advantage in South Jiangsu area gradually weakened, Central Jiangsu area became currently prevailing area of rice yield in Jiangsu Province, and the difference between North Jiangsu area and South Jiangsu area, Central Jiangsu area was decreasing gradually. The proportion of rice output of South Jiangsu area to the whole province declined continuously, and that of North Jiangsu area to the whole province rose obviously. 【Conclusion】Adjustment of planting structure and change of cultivated land area are the influencing factors of regional difference of rice planting area in Jiangsu Province, while adjustment of planting structure is the leading influencing factor. Regional change of rice yield is influenced synthetically by change of cultivated land quality, meteorological factor and social factor. Acceleration of industrialization and urbanization of South Jiangsu area results in the decrease of rice planting area and the weakening of rice yield advantage, which gradually makes the major region of rice production in Jiangsu Province transfer from South Jiangsu area to North Jiangsu area.

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