Scientia Agricultura Sinica

Molecular Identification of Hardness-related Genes of Bread Wheat New Lines in Huanghuai Wheat Region and Their Influence on Yield Characters

ZHANG Fu-Yan, CHEN Feng, DONG Zhong-Dong, SHANG Xiao-Li, CUI Dang-Qun, YIN Gui-Hong

Scientia Agricultura Sinica. 2011, 44(16): 3289-3296. doi:10.3864/j.issn.0578-1752.2011.16.001

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【Objective】 Identification of hardness-related genes in wheat new lines or cultivars developed very recently in Huang-Huai Wheat Region of China could illustrate the distribution of hardness-related genes and provide important gene germplasm for improvement of wheat quality. 【Method】 A total of 109 wheat cultivars or advanced lines widely grown in Huang-huai wheat region were used to identify the phenotype of grain texture and genotype of puroindoline and puroindoline b-2 genes by the technologies of Single Kernel Characterization System (SKCS), PCR amplification with specific primers, restriction enzyme and DNA sequencing. 【Result】 The results showed that hard wheat with percentage of 61.5% was predominant while percentages of the mixed and soft wheats were only 15.6% and 22.9%, respectively, in wheat new lines surveyed. Based on SKCS results, grain hardness index of wheat surveyed ranged widely from 3.2 to 82.6. Three puroindoline alleles of Pinb-D1b, Pinb-D1p and Pina-D1b were detected in hard wheat surveyed. Of them, Pinb-D1b is the most prevalent with percentage of 86.5 %, while Pinb-D1p and Pina-D1b were only 7.5% and 6.0%, respectively. Furthermore, based on identification of puroindoline b-2 gene with gene-specific primers, all surveyed bread wheat genomes D and A contained Pinb-D2v1 and Pinb-A2v4 variants, respectively. However, Pinb-B2v3 variant was only identified in the group B of 86 wheat cultivars or lines surveyed, and Pinb-B2v2 variant was found in the group B of remaining 23 cultivars or lines. The results of analysis of yield characters between Pinb-B2v2 and Pinb-B2v3 variants indicated that the averages of grain number, single grain weight, flag leaf length and flag leaf area of genotypes with Pinb-B2v3 variant were significantly higher than those of genotypes with Pinb-B2v2 variant. 【Conclusion】 Hard wheat was predominant in the wheat germplasms surveyed and Pinb-D1b was the most popular in hard wheat lines. In addition, the wheat with Pinb-B2v3 possessed relatively superior yield character to wheat with Pinb-B2v2.

Comparative Analysis of Genetic Structure and Differentiation of Guangdong and Guangxi Tea Germplasms Based on EST-SSR Markers

QIAO Xiao-Yan, QIAO Ting-Ting, ZHOU Yan-Hua, JIN Ji-Qiang, MA Chun-Lei, YAO Ming-Zhe, CHEN Liang

Scientia Agricultura Sinica. 2011, 44(16): 3297-3311. doi:10.3864/j.issn.0578-1752.2011.16.002

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【Objective】 The well understanding of genetic diversity, structure and differentiation of tea germplasms is important for their effective conservation and reasonable utilization. 【Method】 One hundred and nine core EST-SSR markers were selected and used to analyze the genetic diversity and differentiation of 105 tea accessions from Guangdong and Guangxi provinces. Molecular variance (AMOVA) analysis of inter-groups, inter-populations and individuals within populations was conducted, and then the model-based genetic structure was constructed. 【Result】 A total of 435 alleles were identified with an average of 3.99 alleles per marker (NA). The average of effective number of alleles (NE), Nei's gene diversity (H), observed heterozygosity (Ho), expected heterozygosity (He) and polymorphism information content (PIC) was 2.12, 0.32, 0.59, 0.46 and 0.56, respectively. Guangdong tea germplasms showed lower values of the six indexes mentioned above, compared to Guangxitea. Guangdong Baimaocha (Camellia sinensis var. pubilimba Chang) had lower NA and H values and similar NE, He, Ho and PIC values compared with Guangdong tea (C. sinensis (L.) O. Kuntze). While Guangxi Baimaocha had higher NA value, lower NE, He and H values than Guangxi tea. The values of Fis, Fit and Fst were low among Guangxi and Guangdong Baimaocha and tea populations with strong gene flow. The same results were found between Baimaocha and tea. There were weak genetic differentiation (0.04) and gene flow (6.26) within Baimaocha population. Molecular variance (AMOVA) analysis showed that there was a relatively low level (3.09％) of genetic variation among Guangdong and Guangxi tea germplasms, that within the populations was 94.69%. The population structure analysis could divide all the 105 accessions into 3 groups. Six developed cultivars were grouped into groupⅠ; group Ⅱ consisted of 43 accessions from Guangdong and 16 from Guangxi. The remaining 36 accessions from Guangxi and 4 from Guangdong were assigned into group III.【Conclusion】The genetic diversity of Guangdong tea germplasms was lower than that of Guangxi. Genetic differentiation was insignificant between populations with strong gene flow. Guangdong Baimaocha had much more abundant genetic diversity than Guangdong tea, and genetic differentiation was weak with frequent genetic exchange. Guangxi Baimaocha showed richer genetic diversity than Guangdong Baimaocha, with relatively high genetic divergence and weak gene flow. AMOVA showed that the variation level within Guangdong and Guangxi populations was absolutely higher than among populations.

Effects of Different Furrow-ridge Mulching Ways on Soil Moisture and Water Use Efficiency of Winter Wheat

LI Ru, CUI Rong-Mei, JIA Zhi-Kuan, HAN Qing-Fang, LU Wen-Tao, HOU Xian-Qing

Scientia Agricultura Sinica. 2011, 44(16): 3312-3322. doi:10.3864/j.issn.0578-1752.2011.16.003

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【Objective】The objective of the experiment was to explore the effect of soil water conservation, grain yield and income increments of winter wheat under different furrow-ridge mulching cultivation conditions in semi-humid area . 【Method】 Different cultivation patterns of furrow-ridge mulching were designed using ordinary film, liquid film and straw and soil moisture, yield, water use efficiency and economic benefits of winter wheat were analyzed for three years.【Result】When treated with ordinary film and straw dual-mulching, it had obvious effect of keeping moisture during the whole growth period of winter wheat, which could effectively improve the soil water utilization. In three years, the ordinary film and straw dual-mulching was the highest yield treatment among all treatments, followed by liquid film and straw dual-mulching treatment, the average of yield in three wheat production seasons, increased by 39.3%(P＜0.05) and 29.4% (P＜0.05) compared to conventional flat practice(CK1) respectively, and by 35.6%(P＜0.05) and 25.9%(P＜0.05) compared to the furrow-ridge no-mulching(CK2), respectively. The average of water use efficiency (WUE) of the two increased by 39.6%(P＜0.05) and 30.3%(P＜0.05) compared with CK1, respectively, and by 33.4%(P＜0.05) and 24.5%(P＜0.05) compared with CK2, respectively. The ordinary film and straw dual-mulching gained the highest economic benefit among all treatments and reached up to 4951.4 yuan/hm2 on average in three years, 1452.1 yuan/hm2 higher than CK1, and 1416.5 yuan/hm2 higher than CK2, and was followed by the liquid film and straw dual-mulching, which compared to CK1 and CK2, both showed significant difference（P＜0.05). 【Conclusion】 Under furrow-ridge mulching cultivation conditions, the ordinary plastic film and straw dual-mulching and the liquid film and straw dual-mulching are efficient cultivation patterns for wheat in semi-humid area.

Multi-Spectral Images Estimation Models for Nitrogen Contents of Rape

ZHANG Xiao-Dong, MAO Han-Ping, ZUO Zhi-Yu, SUN Jun, ZHANG Hong-Tao

Scientia Agricultura Sinica. 2011, 44(16): 3323-3332. doi:10.3864/j.issn.0578-1752.2011.16.004

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【Objective】Multi-spectral image analysis method was used to quantitatively analyze the rape total nitrogen content.【Method】The images of rape canopy were taken by the multi-spectral camera and were preprocessed by the median-filtering method. Two-dimensional maximum entropy segment method was used to complete background segmentation of multi-spectral images. 【Result】 By extracting mean and ratio of multi-spectral images of rape canopy , it was found that the features of ARV1, AVS560, ADV1, AVS660 and g are highly correlated with nitrogen content in the whole growth period. Considering the serious multicollinearity between multi-spectral variable, the prediction model of nitrogen content of rape at different growth stages was built by stepwise regression method. 【Conclusion】The reflection intensity distribution information of the visible light and the near infrared light is sufficiently utilized in this research to diagnose the nitrogen content of rape. The multi-spectral image features of the nitrogen content of rape at different growth stages were preliminarily verified. The result shows that the method of multi-spectral image analysis can be used to quantitatively analyze the rape total nitrogen content. This provides a theoretical basis and technical support for the scientific management of rape nutrition.

Analysis of the Source Areas of the Early Immigration of White-Backed Planthopper in Northeast Guangxi of China

QI Hui-Hui, ZHANG Yun-Hui, JIANG Chun-Xian, SUN Ming-Yang, YANG Xiu-Li, CHENG Deng-Fa

Scientia Agricultura Sinica. 2011, 44(16): 3333-3342. doi:10.3864/j.issn.0578-1752.2011.16.005

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【Objective】The objective of this study is to research on the immigratory peak periods and the source areas of the white-backed planthopper (WBP), Sogatella furcifera (Homoptera: Delphacidae), in the northeast Guangxi of China and to provide a basis for early warning and effective control. 【Method】 The Jiaduo light trap was used to monitor the immigratory peak periods of the WBP in Xing’an country, Guangxi from 2007 to 2010. And the Hysplit_4, a software system was used to analyze their migratory trajectory for ascertaining source areas of the WBP. 【Result】 The immigratory peak periods of the WBP in Xing’an country lasted mainly from the mid-May to the end of June. Immigratory abundance of the WBP population in 2007 and 2010 was higher than that in 2008 and 2009. The highest immigratory peak period went back with time from 2007 to 2010. The trajectory analysis showed that the source areas of the WBP were mainly in the north of the Vietnam, and the part of Hainai Island, the south of Guangxi, the southwest of Guangdong. 【Conclusion】 The low level airflow carried large-scale immigration of the WBP from the source areas to the northeast of Guangxi, while heavy rainfall caused mass descent of the migratory population into the areas.

A Nitrogen-Fixing Bacterium Paenibacillus sp. GD812 Antagonistic Against Plant Pathogenic Fungi

CHEN Qian, GAO Miao, HU Hai-Yan, XU Jing, ZHOU Yi-Qing, SUN Jian-Guang

Scientia Agricultura Sinica. 2011, 44(16): 3343-3350. doi:10.3864/j.issn.0578-1752.2011.16.006

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【Objective】 The objective of this study is to screen nitrogen-fixing bacteria antagonistic against plant pathogenic fungi and research nitrogenase activity, nifH, physiological and biochemical characteristics, identification, stress resistance of the functional strains and inoculation effect on plant and to prepare strains for microbial fertilizer production. 【Method】 Nitrogen-free medium was used to isolate nitrogen-fixers and confrontation method was used to screen antagonistic bacteria against plant pathogenic fungus. Nitrogenase activity was determined with acetylene reduction assay. 16S rDNA and nifH were amplified with PCR. Strain identification was carried out based on the morphology, physiology, and biochemical test and 16S rDNA sequence analysis. Microbial inoculation effect on plant was tested with a pot culture in green house. 【Result】 One strain designated as GD812 was selected, which showed nitrogenase activity 30.661 nmol C2H4/h•mg protein, and strongly antagonistic against plant pathogenic fungi Gibberella zeae and Verticillium dahliae with the inhibition rate of 59.5% and 49.3%, respectively. nifH of GD812 is 300 bp sharing 98% sequence identity with that of Paenibacillus sp. Bs57. GD812 was identified as Paenibacillus sp. based on the results of morphology, physiology and biochemical test and 16S rDNA sequence analysis. Further investigations showed that GD812 could use 20 of the 35 tested carbon sources, could grow under temperature ranging from 4 to 50℃, and could grow in range of pH4 to11. Pot culture result showed that fresh weight of Chinese cabbage inoculated with GD812 increased by 52% compared with the control. 【Conclusion】 The newly selected Paenibacillus sp. GD812 showed high nitrogenase activity, strong antagonism against plant pathogenic fungi, extensive carbon source utilization, stress resistance and good inoculation effect, which is a candidate to be further developed for microbial fertilizer production.

Decomposition Characteristics of Rapeseed and Wheat Straws Under Different Rice Cultivations and Straw Mulching Models

WU Ji, GUO Xi-Sheng, WANG Yun-Qing, XU Zheng-Yu, LU Jian-Wei

Scientia Agricultura Sinica. 2011, 44(16): 3351-3360. doi:10.3864/j.issn.0578-1752.2011.16.007

Abstract ( 959 )

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【Objective】The decomposition characteristics and nutrient releasing patterns of rapeseed and wheat straws were studied under different rice cultivations and straw mulching models in 2007 and 2008 growing seasons, respectively.【Method】Wheat and rapeseed straws were wrapped in net nylon bags and put on the surface or embedded into the soil under conventional cultivation and water-saving cultivation models.【Result】The results showed that the decomposition rate of straw was faster in the first 30 days and then slowed down until the end of the experiment. Under water-saving cultivation model, 90 days after straws were embedded into soil, the decomposition rate was high up to 61.06%. The cumulative decomposition rates of wheat and rapeseed straw were 48.88%-59.95% and 50.88%-61.06%, respectively. Under conventional cultivation model, the straws that were mulched decomposed faster than the straws that were embedded into the soil did. With straw mulching, there was no obvious difference in decomposition rates between the two cultivation models. When straws were embedded into the soil, the straw under water-saving cultivation model decomposed faster than the straw under the conventional cultivation model. The sequence of nutrients releasing rates were K＞P＞N≈C. C releasing rates of wheat straw and rapeseed straw were up to 48.29-63.79% and 50.29-66.55%, and N were up to 48.35-52.83% and 46.48%-57.67%, and P were up to 54.83%-67.49% and 56.44%-75.64%, and K were up to 91.98%-95.99% and 92.31%-96.24% after 90 days of decomposition, respectively. The effect of cultivation model and incorporation method on N, P and C releasing patterns had almost the same trend with the decomposition of straw. K release rate was more than 90% after 30 days of decomposition. 【Conclusion】 It is concluded that on the basis of straw mulching, the water-saving cultivation model can promote the straw to decompose and to release the nutrients.

Degradation of 2,4-D by Combined Use of Nanoscale Fe3O4 and Microorganism

WANG Wei-Ping, SI You-Bin

Scientia Agricultura Sinica. 2011, 44(16): 3361-3367. doi:10.3864/j.issn.0578-1752.2011.16.008

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【Objective】 The degradation of 2,4-D with a combination of nanoscale Fe3O4 and microorganism in solution was studied. The research will provide a basis for remediation of pesticide pollution. 【Method】 Nanoscale Fe3O4 would eliminate the toxicity by reductively transform the electron-withdrawing chlorine groups to chloride, then the bacteria was employed to combine with nanoscale Fe3O4 to degrade 2,4-D. The combination mechanism was revealed by analysis of the relationship between bacteria and nanoscale Fe3O4. 【Result】 The results showed that the residual rates of 2,4-D decreased from 100% to 52.0% in 48 h in the presence of 300 mg•L-1 Fe3O4 nanoparticles. Meanwhile, the concentration of chloride ion was increased to 85 μmol•L-1, and the degradation of 2,4-D was a preliminary reductive dechlorination process. 2,4-D could provide carbon sources for promoting microbial growth, and the residual rates of 2,4-D was 57.0% in 5 days by the microorganism treatments. The combination of nanoscale Fe3O4 with microorganism had significantly increased degradation of 2,4-D, and the residual rates of 2,4-D decreased to 35.7% in 7 days, which was much lower than the single treatment of nanoscale Fe3O4 or microorganism. Additionally, microorganisms was also used to degrade 2,4-DCP which was the reductive degradation products of 2,4-D, the residual rates of 2,4-DCP was 50.1% in 5 days, accordingly, the growth OD600 value of microorganisms was 3.29. 【Conclusion】 The degradation rate of 2,4-D by combination treatment was much higher than that separate nanoscale Fe3O4 or microorganism. Nanoscale Fe3O4 could stimulate the growth of microorganisms and reductive degradation product 2,4-DCP was more apt to degradation than 2,4-D.

Construction and Analysis of Eggplant Thermo-Sensitive Parthenocarpy SSH cDNA Library

ZHANG Ying, CHEN Yu-Hui, ZHANG Zhen-Xian, FANG Zhi-Yuan, LIAN Yong, LIU Fu-Zhong

Scientia Agricultura Sinica. 2011, 44(16): 3368-3376. doi:10.3864/j.issn.0578-1752.2011.16.009

Abstract ( 772 )

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【Objective】The aim of this experiment is to investigate the parthenocarpy-related genes in eggplant, and study the parthenocarpic molecular mechanism. 【Method】One forward and one reverse cDNA library were constructed by suppression subtractive hybridization (SSH). All the materials are from the same facultative parthenocapic line D-10, including the parhtenocarpic ovaries and fruits at low temperature, and the unparthenocarpic ovaries and fruits at optimal temperature. The positive clones of the libraries were sequenced randomly, analysed by BLAST， classified by GO and analysed by real-time quantitative PCR. 【Result】A total of 2 347 positive clones in the forward and reverse libraries were obtained. After sequenced and assembled, 1 248 high-quality unigenes were gotten. Blast analysis was made with no redundant database, 1 109 of these unigenes were homologous to known genes, and 139 could be new genes. In Gene Ontology database, 527 unigenes were assigned functional description, and 206, 445, 361 ESTs were, respectively, involved in cell component, molecular function and biological process. According to classification results, the difference between parthenocarpy and unparthenocarpy almost existed in intracellular region. Parthenocarpy may be regulated by some factors and kinases, and most of the differentially expressed genes were involved in basic biological processes.【Conclusion】The SSH cDNA libraries were successfully constructed. Three parthenocarpic related unigenes, including MADS-box gene, pistil extensin-like protein and fruit-ripening gene, were obtained and the possible mechanism was analyzed.

Occurrence Regularity of Mycorrhizal Edible Fungi in the Pine Forests of Beijing Mountain Area

CHEN Qing-Jun, NIU Yu-Rong, HAO Ce, ZHANG Guo-Qing, QIN Ling, LI Jin-Hai, LIU Song, KUI He-Liang

Scientia Agricultura Sinica. 2011, 44(16): 3377-3385. doi:10.3864/j.issn.0578-1752.2011.16.010

Abstract ( 820 )

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【Objective】 The objective of this study was to investigate the occurrence of macrofungi in the pine forests of Beijing mountainous area and their interdependence with the forests, and to provide theoretical data for sustainable exploitation of the pine forests and wild fungi resources. 【Method】 The mycorrhizal edible fungal species, occurrence time, yield, site condition, etc. were analyzed by using 4 years point-survey and temporary investigation methods in pine forests of Beijing mountainous area. 【Result】 It was found that Chroogomphus rutilus and Suillus granulatus are the two most common species of mycorrhizal edible fungi in pine forests, the third high-yielding species is Tricholoma terreum. S. granulatus and C. rutilus usually distribute at 400 to 1 100 meters altitude, August to October, and 4-5 crops in heavy rainfall years. The fruit bodies mature and product spores after about 5 days of their occurrence, and rotted after about one week in high temperature seasons. When the rainfall is enough in spring and autumn, S. granulatus can also appear in early summer and late autumn. After an enough rainfall, about 3 to 5 days of high temperature is necessary for the fungi occurrence. The mycelia of mycorrhizal fungi are very bushy in the soil of mushroom shiro under the pine forests. and the plant species show rich diversity in the shiro area. 【Conclusion】 C. rutilus and S. granulatus are the dominant mycorrhizal edible fungi in the pine forests of Beijing mountainous area. The meteorological factors such as rainfall, temperature are very important for the mushroom occurrence time and yield. With different altitudes and slope positions of pine forests, the occurrence time and dominant species of macro-fungi are also different.

Differential Analysis of Flowering Related Genes by cDNA-AFLP in Chrysanthemum

REN Hong-Yan, SUN Xia, ZHENG Cheng-Shu, WANG Wen-Li, SUN Xian-Zhi, SHU Huai-Rui

Scientia Agricultura Sinica. 2011, 44(16): 3386-3394. doi:10.3864/j.issn.0578-1752.2011.16.011

Abstract ( 849 )

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【Objective】Differentially expressed transcripts of the apical buds in chrysanthemum induced by different photoperiod were analyzed and to isolate the flowering related genes.【Method】cDNA-AFLP technique was used to identify differentially expressed transcript-derived fragments (TDFs) in the apical buds of chrysanthemum induced by different photoperiods of long-day (16h/8h, day/night) and short-day (8h/16h, day/night) treatments, and the TDFs were validated, cloned, sequenced and the bioinformation analyzed. 【Result】 A total of 2 917 TDFs were screened by 64 primer combinations, and 835 TDFs were identified differentially expression, including 584 TDFs up regulation and 251 TDFs down regulation. Fifty TDFs were successfully cloned and the sequence was analyzed. The results indicated that 36 TDFs had homology in NCBI database, and 30 of which displayed homology to genes with known functions, 5 no had a match. Functional analysis indicated that 31 TDFs mainly participated in 7 kinds of processes of signal transduction, development, transcription regulation, protein degradation and synthesis, metabolism, stress responses etc. Semi-quantitative RT-PCR analysis with selected transcripts of 6 flowering related genes indicated that the 6 flowering related genes all differentially expressed or up regulated under the short-day conditions, while they all did not express or down regulate under the long-day conditions. These results were similar to those of cDNA-AFLP expression patterns in chrysanthemum.【Conclusion】In this study several flowering related gene pigments were selected by cDNA-AFLP techniques. These novel genes could be used in future for functional analysis and strategies of molecular breeding.

Development and Application of an IAC-PCR Kit for the Rapid Detection of Salmonella spp.

LI Xiao-Ling, LIU Bin, DAN Xian-Long, WANG Da-Peng, ZHOU Min, SHI Xian-Ming

Scientia Agricultura Sinica. 2011, 44(16): 3395-3402. doi:10.3864/j.issn.0578-1752.2011.16.012

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【Objective】The purpose of this study was to develop a rapid and accurate detection kit for Salmonella spp. using polymerase chain reaction (PCR) with an internal amplification control (IAC). 【Method】 The specific primers and IAC were designed according to the conserved genes invA and stn in Salmonella spp. The optimization of the components and the evaluation of the parameters for the kit were carried out in this study. 【Result】 The experiment indicated that the specific 362 bp DNA fragment was amplified against 147 reference strains of Salmonella spp., while 28 strains of non-Salmonella only showed the 520 bp amplified band of IAC. The detection limit of the kit for purified genomic DNA was 8.0 fg/PCR. It was confirmed in artificial contamination assay that Salmonella spp. could be detected by the kit after 8-10 h enrichment when 4-5 CFU germs were inoculated in 10 mL milk. Thirty food samples were detected by the IAC-PCR kit in this study, and the experiments demonstrated that IAC could successfully indicate false-negative results. Besides, the kit worked well after being frozen-thawed for 60 times or stored at -20℃ for one year.【Conclusion】The IAC-PCR detection kit developed in this study has good performances in specificity, sensitivity, stability and accuracy and is suitable for the rapid and accurate detection of Salmonella spp.

SNPs Screening and Correlation Analysis of ESTs Associated With Bone and Sebum Traits of Pigs

QIN Li-Li, CAI Geng-Yuan, ZHANG Hao, CHEN Yao-Sheng, LI Jia-Qi, WANG Chong

Scientia Agricultura Sinica. 2011, 44(16): 3403-3412. doi:10.3864/j.issn.0578-1752.2011.16.013

Abstract ( 870 )

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【Objective】 QTLs are located in several chromosomes. In order to provide a reference, new candidate genes of bone and sebum traits of pigs were screened.【Method】One hundred and seventy-seven ESTs that may be associated with bone and sebum traits have been searched from NCBI and 21 SNPs were screened by cDNA pool sequencing in 9 ESTs. Four ESTs were located by RH (radiation hybrid). 【Result】 It was discovered that 6 SNPs that are related to bone and sebum traits actually exist in the F2 crossbred population of the Landrace pigs and Lantang pigs after the correlation analysis. E53 is extremely significantly correlated with rear bone (ReB) (P＜0.01)and significantly correlated with bone propotion (BP), fore bone (ForeB) and sebum fat propotion (SFP) (P＜0.05).E82 is extremely significantly correlated with mid-bone (MidB), mid-sebum fat (MidSF) (P＜0.01) and significantly correlated with bone propotion (BP), sebum fat propotion (SFP) and fore sebum fat (ForeSF) (P＜0.05). The differences of E36 in rear sebum fat (ReSF) and fore bone (ForeB), E42 in mid-sebum fat (MidSF) and rear sebum fat (ReSF), E54 in mid-bone (MidB), E87 in fore bone (ForeB) and sebum fat propotion (SFP) reached the remarkable level (P＜0.05). E36 is located on 10q13. E53 is located on 4q21-23. E54 is located on SWC19 of the chromosome 10 without the specific position. E87 is located on 9p21.【Conclusion】E53 and E82 need to be studied further as new candidate genes which affect the bone and sebum traits of pigs.

Development of Equine-Bovine Interspecies Cloned Embryos

ZHU Hua-Bin, SHEN Yan-Jun, DU Wei-Hua, JU Guang-Wei, PANG Yun-Wei, HAO Hai-Sheng, ZHAO Xue-Ming, WANG Dong, WANG Zong-Li

Scientia Agricultura Sinica. 2011, 44(16): 3413-3419. doi:10.3864/j.issn.0578-1752.2011.16.014

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【Objective】The objective of the study was to investigate interspecies somatic cell nuclear transfer (iSCNT) embryonic potential during preimplantation development.【Method】The equine- bovine embryos were reconstructed via hand-made cloning using bovine oocytes as recipient cytoplasm and equine fibroblast as donor cells. Karyotype and the developmental rate were analyzed. 【Result】 Activated with A23187 + 6-DMAP, the iSCNT embryos could develop to blastocyst (2.60%). The cleavage rate and morual rate of cloned embryos cultured in mCR1aa were higher than CR1aa and SOF. Blastocysts were harvested in mCR1aa (2.72%). But there was no difference between CR1aa and SOF. No significant difference was observed in the developmental capacity of cloned embryos and the quality of donor treated with serum starvation or confluence inhibition. The fusion rate of embryos cloned by HMC was higher than microinjection method (90.33% vs 72.94%). Compared equine-bovine iSCNT with bovine–bovine SCNT, there was a significant difference in morual rate and blastocyst rate (21.78% vs 63.52%；2.71% vs 37.48%). 【Conclusion】 It is concluded that the bovine oocytes can support the dedifferentiation and reprogramming of horse somatic cell. However, due to the distance between the two species, the capacity of development of interspecies cloned embryos is far lower than the same species cloned embryos.

Expression of Uterine LHR mRNA During Estrous Cycle in Guinea Pig

JIANG Jin, CAO Shao-Xian, MAO Da-Gan, HUI Feng-Ming

Scientia Agricultura Sinica. 2011, 44(16): 3420-3426. doi:10.3864/j.issn.0578-1752.2011.16.015

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【Objective】 This paper is to clone luteinizing hormone receptor (LHR) fragment from uterine tissue of guinea pig and to analyze evolutionary relationships and the expression characteristics of LHR mRNA during estrous cycle. 【Method】Total RNA extracted from the uterus of guinea pig and primers designed according to the conservative sequence of pig, rat and mice was used for RT-PCR to amplify LHR fragment. Positive clones were identified and sequenced. The DNAMAN software was applied to analyze the homology. ?-actin was used as an inner control and an optimal semi-quantitative RT-PCR method was established to analyze LHR mRNA expression levels in the uterus during 4 estrous cycle phases. 【Result】 Sequencing analysis indicated that there were two kinds of spliced variants: One was the 686 bp fragment and another was 611 bp with a deletion of 75 bp in exon 4. The 686 bp fragment was 83.8% identical to the reported human LHR mRNA, 84.6% to the cattle, 84.2% to the pig, 83.6% to the sheep, 93.7% to the rat, and 99.4% to the mice of LHR mRNA. The uterine LHR mRNA expression was different during 4 phases of the estrous cycle. The levels on day 0(0.635 ± 0.019) and day 4(0.617 ± 0.099) were both significantly lower than that on day 8(P＜0.05), then it rose to the highest on day 12(1.218 ± 0.049) of the cycle. 【Conclusion】 The fragment cloned was a partial sequence of LHR, and the mRNA alternatively spliced variant often existed. The expression level of LHR mRNA in the uterine decreased on days 0 and 4, and increased on day 12. The results have provided a theoretical basis for exploring the functions of LHR in non-gonad tissues of mammals, and meanwhile provided molecular data for further researches in regulation of reproductive endocrinology in other mammals.

Study on Amino Acids Metabolizability and Its Correlation of Evaluation Methods for Adult Roosters of Eight DDGS from Different Resources in China

LI Qiu-Ju, DENG Li-Kang, LI Chao, WANG Zhi-Xiang, ZHAO Qing-Yu, ZHANG Jun-Min

Scientia Agricultura Sinica. 2011, 44(16): 3427-3435. doi:10.3864/j.issn.0578-1752.2011.16.016

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【Objective】A method for rapiding evaluationthe of amino acid metabolizability in DDGS was found to provide scientific basis of rational utilization in DDGS on roosters. 【Method】 45 Hyline brown adult roosters, average body weight (2.6±0.2) kg, were randomly allocatted into 9 groups with 5 replicates per treatment, A 2×9 Youden Square design was carried out for two periods metabolism experiment to determine amino acid metabolizability of 8 DDGS from different sources in China by N-free diet methods and researched the amino acid metabolizability correlation with the DDGS color and water solubles matter. 【Result】 The range of the 8 DDGS total amino acids apparent metabolizability and true metabolizability is 60.18%—74.06% and 74.13%—89.15%, the lysine apparent metabolizability is lower than 60% and true metabolizability is from 63.08% to 82.07%. There is high positive correlation between water solubles matter and lysine true metabolizability (R2 = 0.7601).【Conclusion】The results indicate that there are variations of amino acids metabolizability between DDGS from different sources. Lysine true metabolizability of DDGS on poultry can be estimated by determining their water solubles matter.

Studies on Forage Potential and Mechanism of Maize and Alfalfa Intercropping System

ZHANG Gui-Guo, YANG Zai-Bin, DONG Shu-Ting

Scientia Agricultura Sinica. 2011, 44(16): 3436-3445. doi:10.3864/j.issn.0578-1752.2011.16.017

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【Objective】 The objective of this study were to estimate the discrepancy of forage producing performance and the nutrients digestibility of different maize (Zea mays L.) and alfalfa(Medicago sativa L.) intercropping models, thus to provide a scientific basis for applying the intercropping of maize and alfalfa in practice. 【Method】 Four intercropping treatments, alternating alfalfa and maize rows according to 2﹕2, 3﹕2, 4﹕2, 5﹕2, were designed with three replicates for each. Maize and alfalfa sole crop were the control groups. By determining the forage dry matter and nutrients yields, nutrients digestibility, degradable nutrients yields, and rumen microorganism counts of sheep fed harvested alfalfa-maize mixtures from different intercropping groups, to evaluate the forage production potential and explore the mechanism. 【Result】 The results showed that the yields of dry matter, nutrients and utilizable nutrients of maize and alfalfa intercropping system and alfalfa sole crop were all significantly improved along with the advancing of establishing years (P＜0.05); The yields of dry matter between each treatments had no significant difference (P＞0.05); Compared with the maize sole crop, the intercropping system improved the output and digestibility of crude protein (CP) and the yield of degradable CP, additionally, a linearly response was observed with increasing proportion of alfalfa in intercropping; Compared with the alfalfa sole crop, the intercropping system enhanced the yields and digestibility of ether extract (EE), carbohydrate and gross energy, as well as the yields of corresponding utilizable nutrients; The increase of rumen microorganism counts contributed to the enhanced nutrients digestibility and rendered much more straws tissue cells to be digested in rumen, when the harvested mixtures from intercropping were used as ruminant diets. 【Conclusion】 The alfalfa and maize intercropping system have greater forage nutrients yields than alfalfa or corn monoculture, and improve the digestibility of nutrient and the output of utilizable nutrients, thus it is a viable culturing mode for higher forage production, therefore the result can provide a scientific reference for structuring the optimal ruminant forage producing system in the future.

The Function of PI3K/Akt Signal Pathway During ALV-J Infection in DF-1 Cells

FENG Shao-Zhen, LI Jiao, WU Xiao-Chan, CAO Wei-Sheng, LIAO Ming

Scientia Agricultura Sinica. 2011, 44(16): 3446-3453. doi:10.3864/j.issn.0578-1752.2011.16.018

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【Objective】 Given the biological importance of the PI3K/Akt pathway in virus infection, this experiment investigated whether or not the subgroup J avian leukosis virus (ALV-J) strain HN06 infection in DF-1 cells is correlated with the activity of Akt. 【Method】After inoculation of ALV-J strain HN06 which mainly induces hemangiomas and strain NX0101 which induces myelocytomas in DF-1 cells, respectively, the expression levels of Akt phosphration, viral RNA transcription and viral protein synthesis were analyzed.【Result】 Results showed that the replication rates in vivo of ALV-J strains HN06 and NX0101 were different. HN06 infection led to increased Akt phosphorylation at a very early stage which was virus titer-dependent. LY294002, a PI3K-specific inhibitor, could suppress HN06 induced Akt phosphorylation, indicating that Akt phosphorylation was PI3K-dependent. Moreover, in the presence of LY294002, viral RNA transcription level, viral envelope protein ENV expression and virons secreted in supernatants were suppressed significantly. The PI3K/Akt activation profile and function of HN06 were similar to that of NX0101 reported. 【Conclusion】These data suggest that PI3K/Akt signaling pathway plays an important role in ex vivo ALV-J replication, although the precise mechanism remains under investigation.

The Immunoreaction in Balb/c Mice Immunized with Duck Enteritis Virus gC Genetic Vaccine with Chitosan as Deliver Carrier

JIANG Jin-Feng, CHENG An-Chun, WANG Ming-Shu, LU Li-Ting, ZHU De-Kang, JIA Ren-Yong, CHEN Xiao-Yue

Scientia Agricultura Sinica. 2011, 44(16): 3454-3462. doi:10.3864/j.issn.0578-1752.2011.16.019

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【Objective】 The objective of the experiment is to investigate the influence of chitosan on the immunoreaction induced duck enteritis virus gC genetic vaccine. 【Method】 Balb/c mice were immunized with 50µg per dose of DEV gC genetic vaccine with chitosan as deliver carrier by a single intramuscular injection and oral. Naked plasmid groups of different immunizing doses by intramscular injection, live attenuated vaccine group and physiological saline group were applied as control groups. Cellular immunity, humoral immunity and mucosal immunity were detected, respectively. 【Result】 Compared with DEV gC genetic vaccine with chitosan as deliver carrier by oral and 50 µg of naked plasmid group by intramuscular injection, DEV gC genetic vaccine with chitosan as deliver carrier by intramuscular injection induced stronger cellular and humoral immunity. This vaccine induced similar cellular immune response compared with live attenuated vaccine that induced stronger humoral immune responses. Mucosal immunity was induced only by oral DEV gC genetic vaccine with chitosan as deliver carrier. 【Conclusion】 Chitosan had certain adjuvant’s ability. It has provided a new strategy to obstain more effective DEV gC genetic vaccine. Many factors affecting the immunity efficiency of vaccine need to be optimiazed further for application of this vaccine generally .

Effects of Microbial Fermentation of Corn Straw Mattress Material on Immunological Function of Mice

GAO Jin-Bo, NIU Zhong-Xiang

Scientia Agricultura Sinica. 2011, 44(16): 3463-3468. doi:10.3864/j.issn.0578-1752.2011.16.020

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【Objective】 The aim of this experiment was to study the effect of microbes in fermented corn stalk on immunologic function of mice.【Method】 Forty mice at 4-week-old of age were divided into four groups randomly and raised to establish different kinds of fermention corn stalk with microbes group as well as control group (without addition of microbes). In the experiments, the antibody titer, contents of IL-4 and IFN-γ in serum, the total number of white blood cell (WBC), lymphocyte (LYM) and red blood cell (RBC), and mean corpuscular haemoglobin concentration (MCHC) in blood were determined. The count of Lactobacillus in cecal content and the anti-fatigue time of swimming of mice were examined.【Result】The results showed that microbes in fermented corn stalk significantly promoted the level of antibody and the count of Lactobacillus in cecal content, RBC and MCHC in blood (P＜0.05). The WBC and LYM in blood, the content of IFN-γ, IL-4 were also increased (P＞0.05). The result of anti-fatigue experiment showed that the microbes in fermented corn stalk significantly increased the time of swimming of mice (P＜0.05). 【Conclusion】 Microbes in fermented corn stalk can significantly improve the immune function of mice and other indicators.

Effects of Astragalus, Angelica, Rhodiola on the Growth Performance and Some Physiological Functions of Weaning Piglets

DING Yue-Yun, ZHOU Fen, ZHANG Wei, ZHANG Chen-Hua, YIN Zong-Jun

Scientia Agricultura Sinica. 2011, 44(16): 3469-3476. doi:10.3864/j.issn.0578-1752.2011.16.021

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【Objective】 This study was designed to investigate the effect of astragalus, angelica, rhodiola on growth performance and physiological functions of weaned piglets, and to investigate the mechanism of Chinese traditional medicine on anti-EWS. 【Method】 Total of 180 crossbred (Duroc×Landrace×Yorkshire) weaning piglets (BW=(10.07±0.21) kg), were selected and randomly divided into 10 groups for 2-week-trial. Group 1 was control group fed with basal diet, group 2, 3, 4 were experimental group fed with basal diet plus0.5%, 1%, 1.5% astragalus, group 5, 6,7 were fed with basal diet plus 0.5%, 1%, 1.5% rhodiola, group 8, 9, 10 were fed with basal diet plus 0.5%, 1%, 1.5% angelica. 【Result】 The results showed that: Comparing to the control group, 1)Supplementations with 1%,1.5% astragalus,1.5% rhodiola had the significantly beneficial effects in increasing absolute weight (P＜0.01, P＜0.05, P＜0.05), average daily gain (P＜0.01, P＜0.01, P＜0.01), average daily feed intakes (P＜0.05, P＜0.05, P＜0.05); supplementations with 1% astragalus,1.5% rhodiola had the significantly beneficial effects in decreasing the ratio of feed to gain (P＜0.05, P＜0.05). 2) Supplementations with 1.5% astragalus, 1.5% rhodiola had the significantly beneficial effects in decreasing the content of serum NO on the 7th day (P＜0.01, P＜0.05) and on the 14th day (P＜0.05, P＜0.05), supplementations with 1%, 1.5% astragalus, 1.5% rhodiola and 1.5% angelica had the significantly beneficial effects in increasing the activity of serum GSH-Px on the 7th day (P＜0.01, P＜0.05, P＜0.05, P＜0.05)and on the 14th day (P＜0.01, P＜0.05, P＜0.01, P＜0.05). 3) Supplementations with 1% astragalus, 1.5% rhodiola and 1.5% angelica had the significantly beneficial effects in increasing the red blood cell count (P＜0.01, P＜0.05, P＜0.05), supplementations with 1.5% astragalus had the significantly beneficial effects in increasing the blood hemoglobin content (P＜0.05), supplementations with 1%, 1.5% astragalus, 1.5% rhodiola and 1.5% angelica had the significantly beneficial effects in increasing the white blood cell count (P＜0.01, P＜0.05, P＜0.05, P＜0.05), supplementations with 1% astragalus, 1.5% rhodiola and 1.5% angelica had the significantly beneficial effects in increasing the lymphocytes percentage (P＜0.05, P＜0.05, P＜0.05), and supplementations with 1% astragalus, 1.5% rhodiola had the significantly beneficial effects in increasing the monocytes percentage (P＜0.05, P＜0.05). 【Conclusion】 From above results, it could be concluded that supplementations with astragalus, rhodiola and angelica had the significantly beneficial effects on increasing antioxidant ability, increasing growth performance and increasing physiological functions in weaned piglets, and 1% astragalus had the best effect, 1.5% rhodiola was better, the effect of 1.5% angelica was the least.

Optimization of Solid Fermentation of Botrytis cinerea BC7-3 Strain

ZHANG Li-Hui, FENG Jiang-Lan, DONG Jin-Gao

Scientia Agricultura Sinica. 2011, 44(16): 3477-3484. doi:10.3864/j.issn.0578-1752.2011.16.022

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【Objective】 In order to enhance the production of the herbicidal substances and to furthermore separate the crude toxin, the solid fermenting conditions of BC7-3 were researched in an experiment. 【Method】 BC7-3 was fermented with solid media by using a method of orthogonal combination and different factors affecting toxin-producing was researched. The crude toxin was obtained by extracting with acetone and the herbicidal activity of crude toxin to Flanueria bidentis was bioassayed by using the method of growth inhibition. 【Result】 The medium containing millet at 5 g, sorghum at 5 g, wheat bran at 5 g and corn straw at 2.5 g were found to be the most optimum. The herbicidal activity of the crude toxin reached the strongest when BC7-3 was inoculated with the inoculum concentration of 8% in the optimum medium containing 1% sucrose, 1% ammonium mitrate and 80% water and fermented in the darkness for 15 days under the condition of 20℃. 【Conclusion】 It is suggested that solid fermentation is suitable for the production of the herbicidal substances of BC7-3.

Effect of DA-6 on Enhanced Remediation Efficiency of Solanum nigrum L. in Serious Cadmium Polluted Soil

YU Cai-Lian, LIU Bo, XU Xin

Scientia Agricultura Sinica. 2011, 44(16): 3485-3490. doi:10.3864/j.issn.0578-1752.2011.16.023

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【Objective】 The purpose of this experiment was to study the effect of different concentrations of diethyl aminoethyl hexanoate（DA-6） on the S. nigrum L. biomass and extraction from serious cadmium polluted soil, and discuss the best suitable one which could increase the remediation efficiency. 【Method】 A pot culture experiment was designed by different concentrations of DA-6 with soaking seed and foliage spraying. 【Result】 Results showed that at mature stage, 5, 10, 20 and 40 mg•L-1 DA-6 could increase the shoot dry weight by 8.69%-27.54% (P＜0.05), cadmium concentration by 2.62%-6.75% and total cadmium extraction by 10.12%-36.31% (P＜0.05). MDA got decreased at seedling and mature stages, while SPAD meter value increased; POD activity increased at seedling stage, but decreased at mature stage.【Conclusion】All treatments designed enhanced remediation efficiency, while 20 mg•L-1 DA-6 was the best one.

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