Scientia Agricultura Sinica

Interaction Between AtIPS1 and ATXR5/6 Related to Programmed Cell Death

MENG Ping-Hong, ZHAO Ning, MA Yu-Hua, CHEN Zhi-Lin, DENG Ying, GUO Jing-Tao, ZHANG Xing-Guo

Scientia Agricultura Sinica. 2012, 45(20): 4123-4129. doi:10.3864/j.issn.0578-1752.2012.20.001

Abstract ( 653 )

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【Objective】The objective of the study was to analyse the localization of AtIPS1 and the interaction between AtIPS1 and ATXR5/6 related to programmed cell death (PCD) in Arabidopsis thaliana. 【Method】The constructs of pAtIPS1::AtIPS1-GFP were introduced in BY-2 cells and plants of Arabidopsis. The fluorescence of GFP was observed under a confocal microscope. The interaction between AtIPS1 and ATXR5/6 was analyzed via Bi-Molecular Fluoresence Complementation (BiFC) and the phenotype of mutant atips1 was observed. 【Result】The results showed that AtIPS1 was accumulated in the cytoplasm and the nucleus. AtIPS1 interacted with ATXR5 and ATXR6. The localisation of AtIPS1 was compatible with an interaction with ATXR5/6. The atips1 mutant displayed the phenotype of the spontaneous lesion formation on leaves. The phenotype of atips1 mutant was complemented and no longer formed lesions by a myo-inositol solution treatment or over-expression the AtIPS1 gene. 【Conclusion】This study indicates that the disruption of AtIPS1 results in cell death on leaves. AtIPS1 accumulated in the cytoplasm and nucleus. AtIPS1 could be involved in the regulation of PCD through the interaction between AtIPS1 and ATXR5/6.

Study on Microspore Abortion of Male Sterile Cotton Yamian A and Yamian B

ZHAO Hai-Yan, HUANG Jin-Ling

Scientia Agricultura Sinica. 2012, 45(20): 4130-4140. doi:10.3864/j.issn.0578-1752.2012.20.002

Abstract ( 945 )

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【Objective】 Cell morphological observation and comparison of physiology and biochemistry characteristics of the new male sterile materials Yamian A and its maintainer Yamian B in cotton were studied.【Method】 Yamian A was identified as a new and stable cytoplasmic male sterile line derived from the triple hybrids of G. arboreum, G. bickii and G. herbaceum. This study involved observation of cell morphology and comparison of physiology and biochemicaI characteristics of the new male sterile materials Yamian A and its maintainer Yamian B by means of optical and electron microscopy and measurement of physiology and biochemical indexes.【Result】 Microspore abortion of Yamian A occurred mainly between the stages of sporogenous cell and leptotene. In the microscopic structure, the indications of abortion were that the sporogenous cells were adhesive, the sporogenous cells were without nuclei, then some of them were dissolved at the sporogenous cell stage, the sporogenous cells developed into microsporocyte without going through disintegration, then the nucleus disappeared, and completely disintegrated before the leptotene stage. Compared with the fertile line, the tapetal cell development in the sterile line were delayed, both the tapetal cells and middle layer cells did not collapse in the whole process of anther development. All these abnormal phenomenon resulted in the formation of pollen sacs without pollen grains finally. In the submicroscopic structure, the corresponding tapetal cells also showed an abnormal phenomenon of mitochondrial degradation, while the sporogenous cells and microsporocyte showed degradation signs of mitochondrial cristae, endoplasmic reticulum fracture, and so on. The peroxidase activities in the abortion flower buds of Yamian A were significantly higher than that in its maintainer Yamian B, but its succinodehydrogenase activities were lower than that in its maintainer Yamian B, the superoxide dismutase and cytochrome oxidase activities in the leaves and flower buds of different development stages were both lower than that in its maintainer Yamian B.【Conclusion】 The main cause leading to male sterility of Yamian A may be the delayed development of the tapetal cells, and the changes in the activities of peroxidase, superoxide dismutase, cytochrome oxidase and succinodehydrogenase.

Difference in Root Structure and Respiration Metabolism Between Two Maize Cultivars Under Waterlogging Stress

SENG Shan-Shan, WANG Qun, LI Chao-Hai, LIU Tian-Xue, ZHAO Long-Fei

Scientia Agricultura Sinica. 2012, 45(20): 4141-4148. doi:10.3864/j.issn.0578-1752.2012.20.003

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【Objective】The objective of this study was to identify the difference in root structure and respiration metabolism between two maize cultivars under waterlogging stress in seedling growth.【Method】A pot experiment with two maize cultivars DH662 (a susceptible maize genotype) and XD20 (a tolerant one) was conducted at scientific and educational campus of Henan Agricultural university. CK and 2, 4, 6, 8 days of waterlogging were designed for the two cultivars. The differences in root structure and respiration metabolism between two cultivars were studied. 【Result】The results showed that the root dry weight, root length, root activity of two cultivars decreased with the increase of waterlogging days. And the decreases in DH662 were more significantly than in XD20. The root dry weight in DH662 and XD20 decreased by 29.7% and 12.1%, while root activity decreased by 30.6% and 8.7% as compared with CK treatment after 8 days of waterlogging. The number of aerenchyma, the aerenchyma area and the porosity of two cultivars increased with the increase of waterlogging days. The average aerenchyma area and root porosity of two cultivars increased by 3.9 times and 2.8 times than CK treatment after 6 days of waterlogging. The average activity of alcohol dehydrogenase, pyruvate decarboxylase and lactate dehydrogenase in two cultivars increased by 27.7%, 55.0% and 2.6 times than CK treatment. And the increases in DH662 were more significantly than in XD20. 【Conclusion】Different cultivars had different responses mechanism in their tolerance to waterlogging in seedling growth. A tolerant maize cultivar had intact root structure, higher root activity and developed aerenchyma. And the anaerobic respiration was moderate. So the tolerant maize cultivar could resist waterlogging stress and increase root dry weight.

HgCl2-Induced Programmed Cell Death in Tobacco Cells

LIU Jin-Wei, JIAO Jiao, WANG Lei, WANG Lian-Lian, CHU Xiao-Jing, LIANG Yuan-Cun

Scientia Agricultura Sinica. 2012, 45(20): 4149-4155. doi:10.3864/j.issn.0578-1752.2012.20.004

Abstract ( 855 )

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【Objective】 The objective of this study is to investigate the characteristics of cell death and molecular mechanisms induced by HgCl2. 【Method】 HgCl2-induced cell death was studied in suspension-cultured tobacco cells via neutral red staining, a set of cellular morphological and biochemical characteristics was elucidated in the occurrence of cell death, and pharmacological method was used to explore the mechanisms activated by HgCl2. 【Result】 Within 24 and 48 h, HgCl2 treatment induced cell death in a concentration-dependent manner. Tobacco cells treated with HgCl2 showed typical features of PCD such as chromatin condensation, TUNEL-positive nuclei and DNA ladder formation. HgCl2-induced cell death was accompanied by an increasing production of H2O2. Consistently, the addition of the antioxidants greatly reduced cell death. Pretreatment with protein kinase inhibitor staurosporin, phospholipase C (PLC) inhibitors U73122 and neomycin, and phospholipase D (PLD) inhibitor 1-butanol significantly diminished HgCl2-induced cell death and H2O2 accumulation. 【Conclusion】 HgCl2 induces PCD exhibiting apoptotic-like features. The cell death process requires increased H2O2 production regulated by activation of protein phosphorylation and phospholipid signaling pathways.

Research on Agricultural Data Sharing Based on REST and XML

DAI Jian-Guo, WANG Ke-Ru, LI Shao-Kun, LAI Jun-Chen, XIAO Chun-Hua, LI Shuan-Ming, WANG Qiong

Scientia Agricultura Sinica. 2012, 45(20): 4156-4165. doi:10.3864/j.issn.0578-1752.2012.20.005

Abstract ( 688 )

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【Objective】 Agricultural production management data sharing in the wide distributed and dynamic changing internet environment was studied. 【Method】 REST(Representational State Transfer) as an architectural style was used to solve the problems between systems such as loose coupling, extensible etc. Agricultural production management data structure standards which based on xml and AgroXML were designed to realize unified data understanding in different systems. The prototype system was designed to prove the scheme’s feasibility.【Result】Agricultural production management data was reasonably abstracted as internet resources. The RESTful web services were constructed by WCF(Windows Communication Foundation) based on .net technical platform to realize resource access. Agricultural production management data structure standard was established by improving the schema file of AgroXML to realize data representation unifying, and the data analysis module was developed. The prototype system was developed by using agricultural production management data of a farm of the production and construction corps of Xinjiang as an example to prove the correctness of the scheme, the advantages and disadvantages of the scheme were also discussed, finally, the improvement approach was put forward.【Conclusion】RESTful architectural style can uncouple service provider and client, is suited to wide distributed and dynamic changing environment which contains agricultural production management data, avoiding the problems of traditional distributed technology, meeting the openablity and extensibility of system, and combined with unified data structure standard, which could realize agricultural data sharing between systems.

Determination of Sporulation and Pathogenicity of Ustilaginoidea virens

LI YAN , YU Jun-Jie, LIU Yong-Feng, YIN Xiao-Le, ZHANG Rong-Sheng, YU Mi-Na, CHEN Zhi-Yi

Scientia Agricultura Sinica. 2012, 45(20): 4166-4177. doi:10.3864/j.issn.0578-1752.2012.20.006

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【Objective】 The objective of this study is to test sporulation abilities and steady, pathogenicity, and relativity between the sporulation and pathogenicity of 150 strains of Ustilaginoidea virens. 【Method】A total of 150 strains of U. virens were collected from 9 provinces of China. Their sporulation abilities and pathogenicity were tested by liquid culture and artificial inoculation.【Result】The conidia-producing standard deviation of 132 strains was smaller than 10.0 in three conidia-producing tests, accounted for 88.0% of all strains. Three rice cultivars were inoculated with 150 strains. In the susceptible rice cultivar, Liangyoupeijiu, the average disease index of rice false smut was 67.7, the disease index of 98 strains were more than 50. In the medium resistance cultivar, Huaidao5, the average disease index was 21.8. In the resistant cultivar, Wuyujing3, the average disease index was 16.1, the disease index of 113 strains were shorter than 25. The results indicated that there was a positive relationship between the sporulation and pathogenicity, their correlation coefficient was on the low side, their correlation coefficient was medium-low. According to the pathogenicity of the 150 strains on three rice cultivars, they were divided into 7 pathogenic types. Pathogenic type III (resistant to Wuyujing 3 and Huaidao5, susceptible to Liangyoupeijiu) had 53 strains of U. virens, accounted for 35.3% of all strains, and it was a superior pathogen type. 【Conclusion】The sporulation of U. virens was steady. There was a positive relationship between the sporulation and pathogenicity. There were two interactions between 150 strains of U. virens and three rice varieties with different resistances, one was weak interaction which great majority strains showed, another was strong interaction which few strains showed.

Ecological Effects of Typical Grassland Types in Inner Mongolia on Grasshopper Community

WU Hui-Hui, XU Yun-Hu, CAO Guang-Chun, GE Xi-Ge-Du-Ren, LIU Chao-Yang, HE Bing, E 尔Deng-Ba-Tu, WANG Guang-Jun, ZHANG Ze-Hua

Scientia Agricultura Sinica. 2012, 45(20): 4178-4186. doi:10.3864/j.issn.0578-1752.2012.20.007

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【Objective】The objective of this study is to investigate the ecological relationship between plant community structure and grasshopper community structure in the typical grasslands of Inner Mongolia. 【Method】Canonical analysis, double screening for stepwise regression and redundancy analysis (RDA) were used to analyze the relationship between plant community structure and grasshopper community structure. 【Result】 The biomass loss of Leymus chinensis was negatively correlated with the grasshopper species richness index and positively correlated with the relative abundance of Dasyhippus barbipes in the L. chinensis steppe. The relative abundance of Oedaleus decorus asiaticus was positively correlated with the biomass loss of Stipa krylovii in the S. krylovii steppe and the steppe dominated by S. krylovii with the forb diverse.【Conclusion】In the relationship between plants and grasshoppers，probably the habitat condition consists of structure of plant community can cause more effects on community composition of some grasshoppers than the food plants. The existence of the dominant grasshoppers decided by the dominant plants causes high infestation rate of the dominant plants.

Cloning, Expression Analysis and Binding Characteristics of Chemosensory Protein Gene AlucCSP1 in Apolygus lucorum (Meyer-Dür)

HUA Jin-Feng, ZHANG Shuai, CUI Jin-Jie, WANG Dao-Jie, WANG Chun-Yi, LUO Jun-Yu, LU Li-Min

Scientia Agricultura Sinica. 2012, 45(20): 4187-4196. doi:10.3864/j.issn.0578-1752.2012.20.008

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【Objective】The objective of this study is to clone a chemosensory protein gene AlucCSP1 from Apolygus lucorum, and to test it’s tissue-specific expression and binding characteristics.【Method】The AlucCSP1 was cloned from A. lucorum by RT-PCR. Using semi-quantitative RT-PCR and Real-time PCR methods, the expression pattern of AlucCSP1 in different adult tissues were determined. AlucCSP1 was expressed using pGEX-4T-1 vector and BL21 (DE3) prokaryotic expression system, then purified using GSTrap FF. The binding characteristics of AlucCSP1 with fifty-eight standard odorant samples and five cotton secondary metabolites were investigated using bis-ANS as fluorescence ligand.【Result】The open reading frame full length of AlucCSP1 (GenBank accession: JN573217) was 393 bp, encoding 130 amino acids, including 17 aa of signal peptide in the N-terminal. Sequencing and analysis indicated that AlucCSP1 was characterized by four conservative Cys. It was found that AlucCSP1 was dominantly expressed in the antenna of the female. The molecular weight of recombinant protein of pGEX /AlucCSP1 was about 39 kD. After purification with affinity chromatography and treatment with thrombin protease to cut off GST, the AlucCSP1 protein was obtained. AlucCSP1 had high affinity with gossypol, tannin, quercetin and rutin hydrate, the binding constant was 13.4, 32.7, 19.7 and 38.5 μmol•L-1, respectively .【Conclusion】AlucCSP1 was cloned from A. lucorum and it was dominantly expressed in the female antenna. This may be valuable for studying the role in A. lucorum chemical sensors of AlucCSP1.

Evaluation Method on Soil Fertility Under Long-Term Fertilization

BAO Yao-Xian, XU Ming-Gang, 吕Fen-Tao , HUANG Qing-Hai, NIE Jun, ZHANG Hui-Min, YU Han-Qing

Scientia Agricultura Sinica. 2012, 45(20): 4197-4204. doi:10.3864/j.issn.0578-1752.2012.20.009

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【Objective】To confirm the proper integrated evaluation methods of soil fertility under long-term fertilization.【Method】Based on the data for long-term fertilization at Wangcheng (Hunan province) and Jinxian (Jiangxi province), and the comprehensive evaluation results of the methods of factor analysis, correlation coefficient and Nemoro index were compared and analyzed, and a proper evaluation method was screened out. 【Result】 The results indicated that the integrated fertility indexes(IFI) of factor analysis method and correlation coefficient method showed a similarity in variation trend and difference in different fertilization treatments in over 20 years of long-term experiment, and IFI of Nemoro index method had a difference with the former two and had a high sensitivity. The testing results showed that the interrelations between the IFI of three methods and rice yield showed a very significant positive correction. The result of Nemoro index method had a better relevancy and could well reflect soil fertility to certain extent. The IFI for Nemero index method was as follows: the IFI of low-yield, including non-fertilizer treatment (CK) and partial fertilization treatment (NK), decreased over time. The IFI of high-yield, i.e. balanced fertilization treatment (NPK, 2NPK, NPKCa, NPKS, NPKM), had a small change with time, and the average of IFI for balanced fertilization treatments significantly higher than that of CK and NK treatments.【Conclusion】Factor analysis method, correlation coefficient method and the Nemero index method are suitable for the integrated evaluation of soil fertility under long-term fertilization, but Nemoro index method is the first choice and correlation coefficient method should be selected finally.

Research on Vegetation Response to Temperature and Precipitation in Loess Plateau

ZHANG Chong, REN Zhi-Yuan, LI Xiao-Yan

Scientia Agricultura Sinica. 2012, 45(20): 4205-4215. doi:10.3864/j.issn.0578-1752.2012.20.010

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【Objective】The vegetation response to temperature and precipitation in Loess Plateau was studied.【Method】EOF (Empirical Orthogonal Functions) and SVD (Singular Value Decomposition) were applied to analyze the temporal and spatial response characteristics of vegetation NDVI to the variation of temperature and precipitation, and to distinguish the spatio-temporal correlation between vegetation index and climate factors.【Result】The conditions for vegetation cover in southeast region of Loess Plateau were the best. More precipitation will be favourable to the vegetation cover, including the hydrological effect of Yellow River, but the temperature higher than one threshold value will be less good for plants in arid and semi-arid regions. Higher temperature made plants grow well where there is sufficient moisture in arid region. The vegetation cover has close relations to hydro-thermal condition in valley plain and basin valley. When there is this additional condition, the vegetation cover increases in hydro-thermal equilibrium region. There is a high level of correlation between vegetation cover and hydro-thermal condition, but more limiting factors.【Conclusion】The main limiting factors of temperature on the vegetation cover include land use, vegetation type, cropping system and landform. Loess Plateau is located in drought inland region, so precipitation has direct influence on vegetation cover. Plant growth depends on precipitation in desert zone and non-farming areas. The correlation between vegetation and precipitation is more complicated under irrigation conditions and so on in crop field.

Effects of Exogenous Salicylic Acid on Nitrate Reduction and Assimilation in Chinese Chive Under Weak Light

LI Lin-Yan, WANG Jun-Ling, WANG Mei, GAO Zhi-Kui

Scientia Agricultura Sinica. 2012, 45(20): 4216-4223. doi:10.3864/j.issn.0578-1752.2012.20.011

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【Objective】The effect of exogenous salicylic acid (SA) on accumulation of nitrate, the key enzyme activity of nitrogen metabolism, the apparent electron transport rate (ETR) and the main amino acids and soluble protein was studied in Chinese chive under the weak light in order to understand the effects of exogenous salicylic acid on nitrate reduction and assimilation.【Method】With Chinese chive as an experimental material, two treatment factors including SA (3.0 mmol•L-1) and light intensity (the weak light and natural light) were designed in the experiment. 【Result】Under the weak light, exogenous SA pretreatment reduced the decrease in the key enzyme activity of nitrogen metabolism (nitrate reductase (NR), glufamine synthetase (GS), glutamic- oxaloacetic-transaminase (GOT) and glutamate-pyruvate-transaminase (GPT)), while decreased nitrate accumulation. Compared with no SA treatments，NR and GS activities in SA treatments increased significantly , 23.7% and 12.3%, respectively. Also, the content of chlorophyll (Chl), ETR , the content of free amino acids and soluble protein of weak light were enhanced after SA application. In addition, SA increased the content of most free amino acid components under weak light, with tryptophan and serine as the most effective, reaching 89.8% and 50.6%, respectively, and at the same time reduced the free amino acids and soluble protein ratio (A/P). 【Conclusion】Weak light reduced the capacity of the nitrogen assimilation and material production, however, exogenous SA spraying on leaves obviously improved nitrate reduction and assimilation, which, meanwhile, brought transaminations into play vigorously, and promoted turning NO3- into free amino acid and dissolvable protein. Improvement of nitrate reduction and assimilation might be a key reason for reducing nitrate accumulating into the vacuole of Chinese chive under weak light.

The Effect of Different Nitrogen Concentrations on Fennel Plant Growth, Essential Oil Contents and Components (Foeniculum vulgare Mill.)

HE Jin-Ming, XIAO Yan-Hui, WANG Yu-Mei, PAN Chun-Xiang

Scientia Agricultura Sinica. 2012, 45(20): 4224-4232. doi:10.3864/j.issn.0578-1752.2012.20.012

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【Objective】 The objective of this study was to reveal the response characteristics and the internal correlation of plant growth and essential oil accumulation to nitrogen concentrations. 【Method】 Effect of different nitrogen concentrations on fennel plant growth, essential oil content and components was studied by using the method of solution culture, with Japanese garden formula’s nitrogen content as 1N, and 1/8N, 1/4N, 1/2N, 1N and 2N as designed treatment. 【Result】 With the increase of nitrogen concentration, the plant height, branch number, inflorescence number, number of nodes, shoot fresh weight (FW), fresh and dry weight (DW) of whole plant and yield of essential oil per plant had a trend that firstly increase then decrease, and the maximum values were all in 1N. Total nitrogen, protein nitrogen and pigment contents increased continuously, while total carbon, soluble sugar and essential oil contents and C/N ratio exhibited an opposite trend with increasing nitrogen concentrations. Correlation analysis showed that: total nitrogen content was significantly correlated to protein nitrogen content; total carbon and soluble sugar contents were significantly correlated to C/N ratio; total nitrogen, protein nitrogen contents were significantly and negatively correlated to total carbon content and soluble sugar contents and C/N ratio; essential oil content was significantly correlated with total carbon and soluble sugar contents; yield of essential oil per plant was significantly correlated to total nitrogen, protein nitrogen contents. A total of 23 kinds of components in essential oil were identified, but there were no differences in components of essential oil among different N concentration treatments. However, relative contents of components were significantly different when p<0.01 and p<0.05. (E)-anethole is the dominant essential oil component, contributing 63.89% to 79.28% of the total amount of oil. The second most abundant chemical was limonene, ranging from 5.65% to 18.28% in the total amount of oil, while dill apiol, the third abundant chemical, had the relative amount of 2.26% to 12.61%.【Conclusion】Different nitrogen concentrations not only affect the fennel plant growth, but also affect the yield and concentration of essential oil. Appropriate concentration of nitrogen can contribute to the high rate of plant growth and yield of essential oil.

Effects of Salt Stress on Mesophyll Cell Structures and Photosynthetic Characteristics in Leaves of Wine Grape (Vitis spp.)

QIN Ling, KANG Wen-Huai, QI Yan-Ling, CAI Ai-Jun

Scientia Agricultura Sinica. 2012, 45(20): 4233-4241. doi:10.3864/j.issn.0578-1752.2012.20.013

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【Objective】The changes of cell structures and photosynthetic characteristics in leaves of three grapes, cultivars ‘Cabernet Sauvignon’(Vitis vinifera L.), rootstock ‘5BB’ (Vitis berlandieri × Vitis riparia), and scion/rootstock combinations ‘Cabernet Sauvignon/5BB’, were investigated under salt stress conditions to offer a theoretical basis and technique reference for selecting salt-tolerant genes in grapevine cultivars, rootstocks and scion/rootstock combinations.【Method】In a pot culture experiment, grapevine plants were treated with NaCl at 0 and 100 mmol•L-1 for 30 d. When the height of grapevines was about 60 cm, chlorophyll contents, gas exchange and chlorophyll fluorescence parameters, and cell structure characteristics in leaves were determined by using spectrophotometer, chlorophyll fluorometer, and observed with microscope and transmission electron microscopy.【Results】Compared with the control, the thickness of epidermis cells, palisade tissue and spongy tissue in leaves of grapevine increased, and thickness ratio of the palisade/spongy tissue reduced under salt stress(100 mmol.L-1 NaCl). Thylakoids became swollen, the length and width of chloroplast increased by 130%-150% and 130%-200%, respectively. Meanwhile, chlorophyll (especially Chl b) content, PSII potential activity(Fv/Fo), maximal photochemistry efficiency of PSII (Fv/Fm) and net photosynthetic rate(Pn) dramatically decreased in leaves of grapevines under salt stress(100 mmol•L-1 NaCl). There was a difference in effect on three grapevines by salt stress. Among them, ‘5BB’ showed a lesser negative effect on cell and chloroplast structures, photosynthetic rate, and PSII photochemical reaction. ‘Cabernet Sauvignon’ in the middle and ‘Cabernet Sauvignon/5BB’ was the largest.【Conclusion】The alterations of mesophyll cell structure and chlorophyll content under salt stress(100 mmol•L-1 NaCl) could led to the deduction of photosynthetic electron transfer efficiency and photosynthetic rate. Rootstock ‘5BB’ exhibited higher salt-tolerance characters which may be beneficial to improve the salt-tolerance of ‘Cabernet Sauvignon’ by grafting on the ‘5BB’.

Analysis of Wheat Protein Quality in Main Wheat Producing Areas of China from 2004 to 2011

QI Lin-Juan, HU Xue-Xu, ZHOU Gui-Ying, WANG Shuang, LI Jing-Mei, LU Wei, WU Li-Na, LU Mei-Bin, SUN Li-Juan, YANG Xiu-Lan, SONG Jing-Ke, WANG Bu-Jun

Scientia Agricultura Sinica. 2012, 45(20): 4242-4251. doi:10.3864/j.issn.0578-1752.2012.20.014

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【Objective】In order to understand wheat protein quality status in China in recent years, wheat protein quality in different years and regions was discussed. 【Method】 A total of 4 235 wheat samples collected from the main wheat provinces during 2004-2011 were analyzed for protein content, wet gluten content and sedimentation value, stable time and maximum resistance. 【Result】The protein content and wet gluten content had no obvious change, however, sedimentation value declined slightly from 2004 to 2011; The protein quality in different main wheat provinces was quite different, higher in the north and east places. 【Conclusion】Results showed that emphasis of wheat production on yield has affected the overall quality of wheat seriously in China. Development of high yield wheat varieties with high quality should be focused according to the conditions in different areas.

Effects of the Antioxidant Capacities of 20 Spices Commonly Consumed on the Formation of Heterocyclic Amines in Braised Sauce Beef

YAO Yao, PENG Zeng-Qi, SHAO Bin, WAN Ke-Hui, SHI Jin-Ming, ZHANG Ya-Wei, WANG Fu-Long, HUI Teng

Scientia Agricultura Sinica. 2012, 45(20): 4252-4259. doi:10.3864/j.issn.0578-1752.2012.20.015

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【Objective】The objective of this study was to investigate the effect of spice, which was commonly consumed and possess high antioxidant capacities, to the content of mutagenic and carcinogenic heterocyclic amines (HAs) formed in braised sauce beef.【Method】The total phenols content and antioxidant capacities of aqueous extracts from 20 commonly consumed spices were determined by Folin-Ciocalteu colorimetric method and [2, 2’-azino-bis(3-ethylbenzothiazoline-6-sulfonic acid) diammonium salt] (ABTS) radical cation decolorization assay, respectively. Five spices among them, which contain higher total phenols contents and possess better antioxidant capacities, were further added into braised sauce beef. Contents of HAs were determined by HPLC.【Result】Blank beef sample, which was boiled with only water, contained 9H-pyrido[3, 4-b]indole (Norharman) and 1-methyl-9H-pyrido[3, 4-b]indole (Harman), and their total amount was very low. Beside Norharman and Harman, 2-amino-3, 7, 8-trimethylimidazo[4, 5-f]quinoxaline (7, 8-DiMeIQx), 3-amino-1,4-dimethyl-5H-pyrido[4, 3-b]indole acetate (Trp-P-1) and 3-amino- 1-methyl-5H-pyrido[4,3-b]indole acetate (Trp-P-2) were detected in the control sample containing soy sauce, salt and white sugar, making the total HAs content to 35.72 ng•g-1. Five selected spices, containing clove, galangal, bay leaf, red prickly ash and cinnamon bark, exhibited specific effect on the type and content of individual HA. All of the 5 spices could significantly decrease the contents of Norharman and Harman(P＜0.05), while their effects on the contents of 7, 8-DiMeIQx, Trp-P-1and Trp-P-2 were different. In addition, bay leaf could help the formation of 2-amino-3,4-dimethylimidazo[4, 5-f]quinoline (MeIQ), which was not detected in the control sample.【Conclusion】The water extracts of spices exhibited specific effect on the type and content of individual HA. Generally, red prickly ash and galangal could reduce the total HAs content, significantly inhibit the formation of 7,8-DiMeIQx without generating new HA, and thus exhibit better inhibiting effects.

Evaluation of Bioavailability of Choline in Various Feedstuffs

WANG Si-Jia, CAI Hui-Yi, LIU Guo-Hua, YAN Hai-Jie

Scientia Agricultura Sinica. 2012, 45(20): 4260-4268. doi:10.3864/j.issn.0578-1752.2012.20.016

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【Objective】This study was conducted to evaluate the relative bioavailability (RBV) of choline in corn, soybean meal, canola meal and distillers dried grains with soluble (DDGS). 【Method】A total of 132 one-day-old broilers were divided into 22 groups (6 replicates per group and each replicate had 1 broiler). Groups S1-S6 were standard curve group, Groups A1-A4 were corn groups, Group B1-B4 were soybean meal groups, Group C1-C4 were canola meal groups, and Groups D1-D4 were DDGS groups. The basic diet contained the four feedstuffs above with lower choline content through being extracted with methanol/alcohol. All the test diets had the same composition as that of the basic diet except choline content. A choline response curve was generated using crystalline choline chloride added to the basic diet. The experiment diets were made from the basic diet in which the washed feedstuff was replaced with the intact one with 10%, 20%, 40%, and 100%. In the slope ratio assays to determine choline availability, measured by average daily gain (ADG) and choline concentration in blood.【Result】 Using ADG as response criterion, the choline RBV of corn, soybean meal, canola meal, and DDGS were 63.95%, 104.7%, 37.98%, and 72.09%. Using choline concentration in blood as response criterion, the RBV of corn, soybean meal, canola meal, and DDGS were 94.73%, 184.11%, 23.63%, and 134.23%.【Conclusion】It is possible to evaluate the bioavailability of choline in feedstuffs using ADG and choline concentration in blood as criteria. But the values will be different in different criteria. From high to low, choline RBV of the four feedstuffs were soybean meal, DDGS, corn and canola meal, respectively.

Energy Requirement of Hu Sheep and Dorper Sheep Hybrid F1 Rams

NIE Hai-Tao, YOU Ji-Hao, WANG Chang-Long, WANG Zi-Yu, WANG Feng

Scientia Agricultura Sinica. 2012, 45(20): 4269-4278. doi:10.3864/j.issn.0578-1752.2012.20.017

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【Objective】 This research aimed to define the energy requirement parameters for Hu sheep and Dorper sheep hybrid F1 male rams in the late fattening period, to provide a theoretical basis of scientific supply of energy for this breed of sheep. 【Method】The experiment adopted comparative slaughter method, divided the general requirement into requirement for maintenance and requirement for growth. Adapted metabolizable energy requirement and net energy requirement of these two index systems were used to measure the energy requirements. A total of 42 Hu sheep and Dorper sheep hybrid rams at body weight close to 35 kg were selected, and 30 of them were used in the slaughter experiments. They were randomly divided into 3 intake level groups which were ad libitum group, low-restriction group and high-restriction group, to make their average daily gain reach to 350, 150 and 0g. All rams were slaughtered on 1st day (6n), 20th day (6n) and 42th (18n) day in order to get the retained energy value. Remained 12 rams were used in the digestibility experiment, the rams were divided into 3 intake level groups as comparative slaughter experiment, the recommended formulation was used to predict the methane production and heat production, and then following the above-mentioned results the calculated metabolizable energy intakes of 3 intake levels were 10.69, 11.99 and 12.33 MJ•kg-1 (metabolizable energy intake per kg ration).【Result】The experiment results suggested that the daily net energy requirement for maintenance (NEm) was 271.6 kJ•kg-0.75,the partial efficiency of use of ME for NE for maintenance was 0.66.The daily metabolizable energy requirement for maintenance (MEm) was 413.7 kJ•kg-0.75. Net energy requirements for growth ranged from 0.81-3.12 MJ•d-1 at 35 to 50 kg daily gain 0-400 g. Metabolizable energy requirements for growth ranged from 1.47-5.38 MJ•d-1 at 35 to 50 kg daily gain 0-350 g. The partial efficiency of use of ME for NE for growth was 0.55. 【Conclusion】This research systematicly defined the energy requirement parameters for 35-50 kg Hu sheep and Dorper sheep hybrid F1 male rams in the late fattening period, including net energy for maintenance, metabolizable energy for maintenance, net energy for growth and metabolizable energy for growth, and the results is closely to the domestic and foreign research on energy requirement.

Cloning and Activity Analysis of a Midgut-Specific Promoter BmAPN in Silkworm (Bombyx mori)

LU Gai, CHENG Ting-Cai, JIANG Liang, JIN Sheng-Kai, LIN Ping, HU Cui-Mei, XIA Qing-You

Scientia Agricultura Sinica. 2012, 45(20): 4279-4287. doi:10.3864/j.issn.0578-1752.2012.20.018

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【Objective】The objective of this study is to provide a new tool for the immune-response and application research, and to clone and identify a midgut-specific promoter from Bombyx mori.【Method】 The 5′ upstream regulatory sequence of midgut-specific gene BmAPN (GenBank accession: BAA33715.1) was obtained by PCR from B. mori genomic DNA. A transgenic expression vector named pBac[BmAPN-EGFP-SV40, 3×P3-DsRed] was constructed, in which the reporter gene EGFP was driven by the BmAPN promoter. Furthermore, transgenic B. mori lines was obtained by microinjection and the activity of the promoter at the individual level were detected.【Result】The EGFP driven by a 1 598 bp BmAPN promoter was just specifically expressed in B. mori midgut. It was consistent with the expression pattern of endogenous gene BmAPN. The promoter activity was higher in B. mori larvae than other stages. And the activity in newly exuviated larvae was higher than that of molting larvae. It was speculated that specific components of the promoter might be regulated by hormone.【Conclusion】The BmAPN promoter is an active and midgut-specific promoter in B. mori.

Molecular Epidemiology Study on Porcine Reproductive and Respiratory Syndrome Virus in South Xinjiang

ZHAO Li, LIU Yong-Hong, JIAO Hai-Hong, ZHANG Zhi-Feng, LIU Jun-Feng, CHEN Yan-Zhou, WEN Ya-Qin, LIU Bo, LI Jiang-Tao, CUI Hao-Ran, CHENG Bo, ZHANG Chun-Guang

Scientia Agricultura Sinica. 2012, 45(20): 4288-4299. doi:10.3864/j.issn.0578-1752.2012.20.019

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【Objective】The objective of the study is to determine whether the porcine reproductive and respiratory syndrome virus infection is present in pig farms of South Xinjiang, and to determine the types and gene characteristics of PRRSV strain, thus pertinency effective prevention of PRRS can be made. 【Method】Seven pairs of primers were designed, and by using RT-PCR, the cloning, sequencing and sequence analysis on suspected PRRS pig tissue samples were conducted. 【Result】 There were 35 PRRSV positive in 47 test samples. The positive rate was as high as 74.47% (35/47). The 35 strains were all American type strain. There were 10 strains belong to the highly pathogenic PRRSV, accounting for 21.28% of the total tested samples (10/47), accounted for 28.57% of the positive samples(10/35); According to the comprehensive analysis, no Europe type of strains were found in the samples collected from these pig farms in this study. 【Conclusion】The American type of PRRSV strain infection and highly pathogenic strains exist in South Xinjiang.

Polymorphism Assays of Amino Acid and Establishment of a Two-Temperature PCR for Theileria Annulata Based on Tams1 Gene

LUO Jin, LIU Guang-Yuan, TIAN Zhan-Cheng, XIE Jun-Ren

Scientia Agricultura Sinica. 2012, 45(20): 4300-4309. doi:10.3864/j.issn.0578-1752.2012.20.020

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【Objective】 It has been widespread concern for gene polymorphism assays in species classification, pathogen detection and vaccine screening. T. annulata is an important blood protozoa in bovine and causes major hazards. In present study the Tams1 amino acid polymorphism of T. annulata was analyzed and a two-temperature PCR detection method was established for T. annulata. 【Method】The specific primers were designed of Tams1 gene of T. annulata. A nucleotide fragment of 846 bp in length was obtained by PCR amplification. The gene amino acid sequence was compared and analyzed with 12 known species in different regions of the isolates in GenBank. And other primers were designed in conserved region of Tams1 gene and the detection method of T. annulata was established by two-temperature PCR. The method was used to detect theileriosis in field.【Result】The fragment encoded 281 amino acids, including 48 basic amino acids, 42 acidic amino acids and 100 hydrophobic amino acids. Identity analysis showed that Gansu strain of T. annulata had closest relationship with Ankara (Z48739.1), Turkey (AF214911), Bahrain (AF214794) and had little relationship with Xinjiang strain (No. accession number) while had closest relationship with Italy (AF214862) and Spain (AF214815) strains. The results can be testified by sequences alignment. The two-temperature PCR detection method can detect in 0.31 fg•μL-1 blood infection. Specificity results showed that only T. annulata genome DNA test was positive, other control parasite genome template were shown to be negative, and no cross reaction with other infected bovine Piroplasmorida. The two-temperature PCR method of T. annulata was used to test 335 field samples, the positive rate was 16.33%, and the microscopic detection results of 2.25% coincidence rate was 100%. Compared with common PCR, the method had no significant difference in sensitivity. But, two-temperature PCR was higher than common PCR in specificity, and the method took a short time with repeated heating and cooling.【Conclusion】T. annulata Tams1 genes from different strains exist obvious differences. Its amino acid sequence polymorphism feature is significant. Therefore, when the gene is used as a potential candidate antigen attention should be paid to the design of peptide vaccines. Two-temperature PCR method has good sensitivity and specificity. It has significance for early detection, early prevention of T. annulata.

Effects of Planting Methods on Nitrogen Accumulation and Distribution of Mid-Late Indica Hybrid Rice Combination ⅡYou 498 Under Different Ecological Conditions

DENG Fei, WANG Li, REN Wan-Jun, LIU Dai-Yin, YANG Wen-Yu

Scientia Agricultura Sinica. 2012, 45(20): 4310-4325. doi:10.3864/j.issn.0578-1752.2012.20.021

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【Objective】 The hybrid rice combination II You 498 was used to clarify the effects of ecological conditions, planting methods and their interaction on nitrogen accumulation, distribution and production efficiency of rice in Sichuan province. 【Method】 Field experiments were conducted in Renshou, Pixian and Ya’an in 2011, involving two seedling ages and four transplanting methods. During rice growth period, the dry matter weight, yield and nitrogen content of organs were measured, and the nitrogen accumulation, distribution, translocation and production efficiency under different ecological conditions were analyzed. 【Result】The nitrogen content and accumulation of rice decreased, while the nitrogen use efficiency and nitrogen harvest index significantly increased in Renshou which were characterized by low soil fertility and high light intensity. The 50-day-old seedling transplanting treatment improved the nitrogen accumulation during jointing-heading period and translocation of rice leaf and stem-sheath nitrogen after heading stage. Furthermore, it was found that the single seedling optimized-broadcasting treatment enhanced the nitrogen accumulation rate of rice during sowing-jointing and heading-maturity periods and thus improved the total nitrogen accumulation. According to the correlation analysis, rice yield in Renshou was positively and significantly correlated to the nitrogen accumulation during sowing-jointing period and the translocation of rice leaf nitrogen, but the rice yield in Pixian was positively correlated to the nitrogen accumulation during heading-maturity period. In Ya’an, the effective panicle of rice was correlated to the nitrogen content of plant at maturity stage, while the spikelet number to the translocation of stem-sheath nitrogen, positively and significantly. 【Conclusion】In order to coordinate the accumulation and distribution of nitrogen and increase the total nitrogen accumulation and grain yield, a suitable planting method, such as 50-day-old single seedling optimized-broadcasting treatment in Renshou and Pixian, 50-day-old single seedling hand-transplanted treatment in Ya’an, must be chosen to adapt different ecological conditions.

Inheritance and Gene Cloning of an ALS Inhabiting Herbicide- Resistant Mutant Line M9 in Brassica napus

HU Mao-Long, PU Hui-Ming, GAO Jian-Qin, LONG Wei-Hua, QI Cun-Kou, ZHANG Jie-Fu, CHEN Song

Scientia Agricultura Sinica. 2012, 45(20): 4326-4334. doi:10.3864/j.issn.0578-1752.2012.20.022

Abstract ( 777 )

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【Objective】The objective of this study is to identify and characterize the resistant gene of a mutant line M9，which was previously found in the rapeseed (brassica napus L.) by spontaneous mutation, conferring resistance to acetolactate synthase or acetohydroxyacid synthase (ALS or AHAS) inhibiting herbicides, and understand the molecular basis for resistance to ALS-inhibiting herbicides.【Method】Reciprocal crosses were made in order to study the inheritance of resistance. Three genes BnALS1-BnALS3 encoding ALS were isolated from the mutant and wild type by using the homology-based candidate gene method. The resistant gene of M9 was transformed into Mutsu-Isuzu cytoplasmic male sterile (MICMS) restorer lines by hybridization, microspore culture, and PCR analysis. 【Result】The resistance of M9 was inherited as a single, dominant nuclear gene. Three genes BnALS1-BnALS3 encoding ALS were isolated. Molecular analysis identified a single-point mutation leading to an amino acid substitution from serine 638 (AGT) to asparagine (AAT) at the herbicide-binding site of the rapeseed BnALS1 gene. All the resistant restoring lines had the specific band of BnALS1R by sequencing. 【Conclusion】The resistance trait of M9 is controlled by a single dominant nuclear gene. The molecular basis for the resistance to ALS-inhibiting herbicides results from the point mutation (Ser638Asn).

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