Loading...

Table of Content

    16 April 2016, Volume 49 Issue 8
    CROP GENETICS & BREEDING·GERMPLASM RESOURCES·MOLECULAR GENETICS
    Growth and Lipid Accumulation Promotion of Chlorella by Endophytic Pantoea sp. from Rice Seeds
    SHI Yu-qian, ZHAO Yan
    Scientia Agricultura Sinica. 2016, 49(8):  1429-1442.  doi:10.3864/j.issn.0578-1752.2016.08.001
    Abstract ( 537 )   HTML ( 2 )   PDF (1166KB) ( 733 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this study is to isolate the dominant endophytic bacterial strain from rice seeds and identify its effects on the growth, biochemical composition and lipid accumulation of Chlorella, to explore the feasibility of plant endophytes as new microbial resources for growth promotion and biochemical quality improvement of microalgae. 【Method】The endophytic bacteria was isolated from rice seeds by the traditional surface sterilization method, using rice (Oryza sativa L. japonica. cv. Nipponbare) seeds which were harvested from the paddy fieldas experimental materials. Identification of the endophytic bacteria was based on morphological observation and 16S rDNA sequence analysis. Using Chlorella pyrenoidosa as the experimental subject, effects of the endophytic bacteria on the growth and biochemical composition of Chlorella were evaluated by co-cultivation method. The algal cells were co-inoculated with endophyte at the number ratio of 1:10 in BG11 medium with axenic C. pyrenoidosa as control. The algal cell number was determined by monitoring the cell count using hemocytometer. Meanwhile, the change curve of the concentration of bacteria in bacterial-algae co-cultivation was determined by the colony form counting method with flat plate dilution for single bacterial culture as the control. The algal chlorophyll a and carotenoids were extracted with methanol and the contents were determined by UV spectrophotometry. The algal total lipids were extracted with chloroform/methanol and the fatty acid composition was analyzed by gas chromatography/mass spectrometry. 【Result】A endophytic bacterial strain was isolated and named as REY-1, which is very commonly distributed in rice seeds with larger number. The REY-1 is Gram-negative, short rod in cell-shaped. The REY-1 strain belongs to Pantoea sp. according to the 16S rDNA identification. After 14 days co-cultivation of the REY-1 and C. pyrenoidosa, the algal cell concentration in bacterial-algal co-cultivation reached 1.15×108 cells/mL that was 1.97 times as many as the control. The concentration of REY-1 in bacterial-algal co-cultivation and single bacterial culture both had peak valley changes curve, but the concentration of bacteria in bacterial-algal co-cultivation was significantly lower than that of single bacterial culture at the same period. The ratio of bacterial concentration to algal concentration decreased from 10:1 to 1:100 at the end of incubation. The result showed that REY-1 can promote the growth of C. pyrenoidosa, but its proliferation was inhibited by algae at the same time, and there was a significant interaction effect between the two. The total concentration of chlorophyll a and carotenoids in the co-cultivation system had no significant difference with the control, whereas the decrease of chlorophyll a and carotenoid contents in single algae cell was significant at 42.58% and 42.68% compared with the control, respectively. The total lipid content of algae in co-cultivation reached 29.90% by a significantly increase of 78.00% compared with the control, and the lipid productivity (2.14 mg·L-1·d-1) was 1.68 times higher than the control. Palmitic acid (16:0), linoleic acid (18:2) and linolenic acid (18:3) are major fatty acids in algae cells in both cultivations. Co-cultivation treatment with REY-1 increased the relative content of monounsaturated fatty acids and short chain myristic acid (14:0) in algae cells by 32.37% and 2.12 times, respectively, compared with the control, which reached to significant or very significant levels. In addition, a new long chain fatty acid, erucic acid was synthesized in co-cultivation algae cells, with relative content reached 1.68% of that of the algae total fatty acid. 【Conclusion】The endophytic Pantoea sp. REY-1 isolated from rice seeds can survive in long-term in BG11 liquid medium and interact with C. pyrenoidosa. It can significantly lead to a cumulative increase in biomass and lipid content of C. pyrenoidosa, and change its relative composition content of fatty acids. It can also promote the biosynthesis of short and long chain fatty acids. Rice seeds endophytic bacteria can be used as a good new microbial resource to develop beneficial symbiotic bacteria for Chlorella sp.
    Mapping QTL for Plant Height and Ear Height in Maize Under Multi-Environments
    HE Kun-hui, CHANG Li-guo, CUI Ting-ting, QU Jian-zhou, GUO Dong-wei, XU Shu-tu, ZHANG Xing-hua, ZHANG Ren-he, XUE Ji-quan, LIU Jian-chao
    Scientia Agricultura Sinica. 2016, 49(8):  1443-1452.  doi:10.3864/j.issn.0578-1752.2016.08.002
    Abstract ( 704 )   HTML ( 13 )   PDF (4585KB) ( 867 )   Save
    References | Related Articles | Metrics
    【Objective】QTL mapping for plant height and ear height were conducted with phenotype data collected from multi-environments, in order to identify stable QTL for plant height and ear height, which will provide a theoretical basis for molecular breeding of ideal plant type in maize.【Method】A total of 150 F7 recombinant inbred lines (RILs) population as the experimental materials were derived from the dominant maize inbred lines Xu178 and K12. Firstly, selecting 495 SSR markers from the MaizeGDB to detect polymorphism in parents, using markers with polymorphism to analyze population genotype. MapMaker V3.0 was used to part the linkage groups and to construct the linkage map. Secondly, QTL mapping for plant height and ear height were detected with inclusive composite interval mapping (ICIM) method of QTL IciMapping V4.0. Using phenotype data which were collected in two years and from three locations (Yulin, Shaanxi. Yangling, Shaanxi. Huludao, Liaoning. 2014-2015) and estimated the breeding value. Finally, conditional QTL analysis was conducted and compared with unconditional QTL analysis to discuss the genetic relationship between plant height and ear height at the QTL level.【Result】The genetic map was constructed with 191 filtered SSR markers, and the total length was 2 069.1 cM with the average length 10.8 cM. For plant height and ear height from 6 environments and estimated breeding value, ten plant height QTL and eight ear height QTL were detected, which distributed on the chromosomes 1, 3, 4, 5, 6, 7, 8 and 10, respectively, the LOD values ranged from 3.25 to 8.36, the additive effect from -6.41 to 8.70, and the range of individually explaining phenotypic variation was from 6.96% to 27.41%. Above of all QTL, six major QTL with LOD more than 10.00% could be detected in 3 or more environments. Of which, two QTL were detected on the same region of Bin5.01/5.02 in 6 environments, the LOD values ranged from 3.25 to 6.48, the additive effect from 4.05 to 8.70. Two QTL were detected on the same region of Bin3.03/3.04 in 5 environments, the LOD values ranged from 4.71 to 8.36, the additive effect ranged from 4.93 to 6.36. As well as two QTL were detected on the same interval of Bin6.02 in 3 environments, the LOD values ranged from 3.52 to 5.21, the additive effect ranged from 4.38 to 8.16, respectively. The favorite alleles of these QTL were all coming from female parent Xu178. Conditional analysis and unconditional analysis showed that the six QTL of three chromosomal regions were three pleiotropism loci which controlled plant height and ear height.【Conclusion】The inheritance of plant height and ear height of maize is strongly influenced by the environments, most of the QTL can be detected only in 1 or 2 environments. Three major QTL can be detected in 3 or more environments, and they are able to stable heredity with high phenotypic variation, it is expected that the major QTL could be applied in the molecular breeding.
    TILLAGE & CULTIVATION·PHYSIOLOGY & BIOCHEMISTRY
    Effects of Straw Management and Sowing Methods on Soybean Agronomic Traits and Soil Physical Properties
    WANG Xing, WU Cun-xiang, QI Yu-jun, XU Ze-jun,WANG Zong-biao, HAN Tian-fu
    Scientia Agricultura Sinica. 2016, 49(8):  1453-1465.  doi:10.3864/j.issn.0578-1752.2016.08.003
    Abstract ( 507 )   HTML ( 1 )   PDF (571KB) ( 559 )   Save
    References | Related Articles | Metrics
    【Objective】 This study is to evaluate the effects of wheat straw management and sowing methods on soybean growth and development and soil physical properties in the wheat-soybean double-cropping system of Yellow-Huai-Hai Rivers Valley, to illustrate the key points of no-till sowing technologies, and to assess the utilization value of a new method of no-tillage plus straw mulching and precise sowing (NTSMPS), in order to provide the technical solution for improving summer soybean productivity in this region. 【Method】The split-plot field experiment, in which the main-plot factor was the sowing method while the sub-plot factor was straw-returning amount, was performed in Xuzhou, Jiangsu Province, China, in 2012-2014. The sowing method factor had three levels: NTSMPS, conventional no-tillage plus-mechanical drilling (CNSD), and rotary-tillage plus artificial broadcasting (RTAB), and the straw returning amount had two levels: Total-amount and half-amount straw returning. With six treatments, we compared the effects of straw management and sowing methods on soybean sowing and emergence quality, growing process, yield, soil physical properties and wheat straw decomposition rate. 【Result】 The emergence date of NTSMPS was 1 day and 1-3 days earlier than that of CSND and RTAB, respectively. Meanwhile, the emergence rate of NTSMPS was the highest among the three treatments. Furthermore, the date of beginning bloom (R1 stage) of NTSMPS was 0.1-2.3 days earlier than that of the other two sowing methods, and this difference was significant (P<0.05). However, straw returning amount had no effect on emergence speed and growth rate. The yield in the NTSMPS was 4.12%-12.40% and 1.96%-5.50% higher in a comparison with that of CSND and RTAB, respectively. In contrary, there was insignificant difference of yield among two straw returning amount levels. Compared with RTAB, NTSMPS and CSND resulted in an obvious reduction in soil temperature and increase in soil moisture during sowing and seedling emergence (P<0.05). Soil humidity was higher in treatment with total-amount straw application than in treatment with half-amount straw returning, while soil temperature was lower only in the dry year (2012). Wheat residue decomposition ratio of RTAB was lower as compared to NTSMPS and CSND. However, wheat straw of NTSMPS exerted no hazardous influence on the sowing and growth of the next season crop (winter wheat).【Conclusion】Straw mulching is a good way for wheat residue management in wheat-soybean double cropping system because of its functions in soil moisture remaining, temperature adjustment, soil sealing prevention, soybean emerging enhancement, and organic matter and mineral supply for the next-season wheat. NTSMPS technology is powerful to synchronously resolve the sticky questions of wheat residue management, and soil fertility improvement and can be widely used in the Yellow-Huai-Hai River Valley and similar areas.
    Effects of Shade Degrees on Photosynthesis and Lodging Resistance Degree of Different Shade Tolerance Soybean
    LIU Ting, LIU Wei-guo, REN Meng-lu, DU Yong-li, DENG Yu-chuan, ZOU Jun-lin, FANG Ping, YANG Wen-yu
    Scientia Agricultura Sinica. 2016, 49(8):  1466-1475.  doi:10.3864/j.issn.0578-1752.2016.08.004
    Abstract ( 572 )   HTML ( 11 )   PDF (407KB) ( 645 )   Save
    References | Related Articles | Metrics
    【Objective】Under the maize/soybean relay strip intercropping system, shading from high crop maize leads to soybean seedling thinning, lodging and decreasing soybean yield. We studied the effect of soybean photosynthesis and lodging resistance in seeding stage under different shade degrees in the maize/soybean relay strip intercropping system, then provided a theoretical basis for the establishment of a reasonable community population structure. 【Method】 The non-shade tolerance and non-lodging resistant Nan032-4 and the strong shade tolerance and strong lodging resistance Nandou 12 were taken as experimental materials in 2014 and 2015. We designed three light conditions in the outdoor pod experiment, the natural light (W0), one layer of black sun-shade net (W1), and two layers of black sun-shade net (W2) with light transmittances of 100%, 32% and 11% respectively. Photosynthetic characteristics such as soybean photosynthetic rate, stomatal conductance, transpiration rate, intercellular CO2 concentration,SPAD values, leaf area and biomass accumulation of each organ and lodging characters, including plant height, each internode length, each internode diameter, stem snapping resistance, lodging resistance index were measured and analyzed. 【Result】 The results showed that as the shade increased, photosynthetic rate, stomatal conductance, leaf area and SPAD values, the dry matter accumulation of each organ and root/shoot ratio of the two soybean varieties were all reduced significantly. As shade increased, seedling lodging became serious, with the stem snapping resistance and lodging resistance index also reduced significantly. However, the shade avoidance responses on different shade degrees were significantly different between these two different shade tolerant varieties. Compared with W0, photosynthetic rates of the shade tolerant cultivar Nandou12 and the non-shade tolerance Nan032-4 under W1 and W2 were decreased by 35.6%, 99.6% and by 40.9%, 83.6% respectively. Compared with Nandou12, Nan032-4 was influenced more seriously. The dry matter accumulation of Nandou12 under W0 and W1was higher than Nan032-4, but under W2 there was no significance between the two varieties (P<0.05), this indicated that the photosynthetic rate of soybean remained at a higher level under moderate shade and the photosynthetic product of the shade tolerant cultivar was higher than the non-shade tolerance cultivar. The lodging rate over 2014 and 2015 of Nan032-4 under W1 and W2 was higher than that of Nandou12 by 55.9% and 2.57%. Under the treatment W1, Nan032-4 showed moderate lodging and Nandou12 showed mild lodging; Nan032-4 while Nandou12 showed serious lodging under the treatment W2. Under the same shade level, the snapping resistance of Nandou12 (39.8%) was higher than Nan 032-4 (38.4%), but the snapping resistance of Nandou12 under W1 and W2 for 2014 and 2015 was higher than that of Nan032-4 by 27.3% and 26.6%. Under the same shade level, stem snapping resistance and lodging resistance index of the shade-tolerant cultivar Nandou12 were much higher than that of Nan032-4 (P<0.05).【Conclusion】With increasing shade level, photosynthetic accumulation of soybean was reduced and lodging was more serious. The two materials showed the shade sensitive under the excessive shade. But under moderate shade, the strong shade tolerant cultivar maintained its high photosynthetic and lodging resistance. In a word, under the maize/soybean relay strip intercropping system, we should appropriately reduce the shading stress on soybean in production, then finally realize soybean potential of shade tolerance and lodging resistance.
    PLANT PROTECTION
    Progress in Research on Mechanism of Resistance to Fusarium Head Blight in Wheat
    LIU Yi-ke, TONG Han-wen, ZHU Zhan-wang, CHEN Ling, ZOU Juan, ZHANG Yu-qing, GAO Chun-bao
    Scientia Agricultura Sinica. 2016, 49(8):  1476-1488.  doi:10.3864/j.issn.0578-1752.2016.08.005
    Abstract ( 923 )   HTML ( 23 )   PDF (442KB) ( 1245 )   Save
    References | Related Articles | Metrics
    Fusarium head blight (FHB) is one of the most important diseases of wheat worldwide, which poses a serious threat to wheat production and food safety. According to the form of wheat resistance to FHB, the resistance types can be divided into five categories: resistance against initial infection (Type I), resistance to pathogen spreading in infected tissue (Type II), resistance to kernel infection(Type III), tolerance(Type Ⅳ) and resistance to toxins in ears by decomposing them(Type V). Resistance mechanism types are usually classified into either morphological or physiological, and the morphological resistance mechanism is synonymous with passive. Wheat morphological characteristics including plant height, heading date and flowering duration, anther extrusion, presence or absence of awns, ear length and density, flower opening width and waxy surfaces on ear tissue may be related to resistance against initial infection. Cytological studies showed that the resistant varieties infected by pathogen can make rapidly defense response by the cell structure, physiological and biochemical responses. The formation of thick-layered appositions and papillae and the increase of lignin, thionins, hydroxyproline-rich glycoproteins and hydrolytic enzymes are cooperated to resist the pathogen development in the resistant varieties. In the complex plant signaling pathways, three signaling pathways including salicylic acid (SA), jasmonic acid (SA) and ethylene (ET) play the most important role in the antifungal response of plant. The role of SA and ET signaling pathways in wheat resistance to FHB is still in controversy, and the positive role of JA signaling pathway has been confirmed by most of the researchers. So far, it is found that more than 200 FHB resistance quantitative trait loci (QTLs) are distributed on all 21 wheat chromosomes, among them 22 QTLs were mapped by different mapping populations, including two stable Type I sites Fhb1 and Fhb2 locating on chromosome 3BS and 6BS, respectively, and two stable Type II sites Fhb3 and Fhb4 locating on chromosome 4B and 5A, respectively. When infected by the plant will produce a series of complex signaling pathway to activate resistant response, and induce the expression of defense-related genes, thereby causing protein and metabolic changes to resist the invasion of pathogen. Studies have indicated that the pathogenesis-related genes, antimicrobial peptide genes, transcription factor genes, detoxification-related genes and other FHB resistance genes are involved in the process of wheat resistance to FHB. In the future, major genes controlling FHB resistance can be cloned by map-based cloning technology, and based on genome-wide association studies and various omics techniques, the mechanism of wheat FHB resistance will be studied on whole genome and gene regulatory network level. This paper can provide a reference for the related research on resistance mechanism to FHB in wheat.
    Function of Polygalacturonase Genes Vmpg7 and Vmpg8 of Valsa mali
    XU Chun-jing, WU Yu-xing, DAI Qing-qing, LI Zheng-peng, GAO Xiao-ning, HUANG Li-li
    Scientia Agricultura Sinica. 2016, 49(8):  1489-1498.  doi:10.3864/j.issn.0578-1752.2016.08.006
    Abstract ( 517 )   HTML ( 6 )   PDF (1445KB) ( 545 )   Save
    References | Related Articles | Metrics
    【Objective】Transcriptome analysis showed that Vmpg7 and Vmpg8 are two significantly up-regulated polygalacturonase genes of Valsa mali during the pathogen infection progress. The objective of the study is to explore the role of these two genes in pathogenicity and lay the foundation for further pathogenesis of V. mali by analyzing the vegetative growth, pathogenicity, pectinase activity, the utilization of pectin, and the effect of gene knock-out of Vmpg7 and Vmpg8 to the expression levels of the other PG genes in PG family.【Method】qRT-PCR was used to detected the expression levels of Vmpg7 and Vmpg8 during the pathogen infection progress and the expression levels of other PG genes in PG family when Vmpg7 and Vmpg8 were deleted. The single and double deletion mutants were constructed and confirmed using double-joint PCR, PEG-mediated protoplast transformation technique, PCR with four pairs of primers and Southern blot, and the gene complemented mutants were constructed by gap repair technology. The PDA routine culture was selected to analyze the vegetative growth of mutants. In vitro inoculation to apple leaves and twigs was used to detect the pathogenicity. The 3,5-dinitrosalicylic acid (DNS) colorimetric method was used to detect the activity of extracellular pectinase, and the Czapek culture medium was selected to observe the utilization of pectin.【Result】The expression levels of Vmpg7 and Vmpg8 were assayed by qRT-PCR, and results showed that the two genes were up-regulated 27.73 and 8.19 folds at 3 days after infection, respectively. With the help of the gene knockout technique, three deletion mutants of Vmpg7, one deletion mutant of Vmpg8, three deletion mutants of Vmpg7/Vmpg8, together with the complemented mutants of Vmpg7 and Vmpg8 were constructed, respectively. When the mutants were cultured on PDA medium, the colony morphology and growth rate showed no significant change. When they were inoculated to apple leaves and twigs, the pathogenicity of the Vmpg7 deletion mutant and the Vmpg7/Vmpg8 double deletion mutant showed significant reduction, and the pathogenicity of the Vmpg8 deletion mutant was only reduced on apple leaves. Further analysis of pectinase activity and the utilization of pectin showed that the pectinase activities of the Vmpg8 deletion mutant and the Vmpg7/Vmpg8 double deletion mutant were significantly reduced, and all the mutants had a slow growth rate on pectin medium. Interestingly, the pathogenicity, pectinase activity, and the growth rate on pectin medium were back to the level of wild-type 03-8 when the genes were complemented back to the distinct mutants, respectively. More importantly, the expression levels of the other PG genes in PG family were obviously affected when Vmpg7and Vmpg8 were knockout, especially three genes that were significantly up-regulated.【Conclusion】Vmpg7 and Vmpg8 participate in pathogenic process of V. mali by adjusting the pectinase activity, which may synergized with the other PG genes in the same family.
    Duplex-PCR Detection for Heterodera avenae and Heterodera filipjevi Based on mtDNA-COI Sequences
    NIU Wen-wen, WANG Xuan, LI Hong-mei, JU Yu-liang, WAN Wen-wen
    Scientia Agricultura Sinica. 2016, 49(8):  1499-1509.  doi:10.3864/j.issn.0578-1752.2016.08.007
    Abstract ( 356 )   HTML ( 2 )   PDF (692KB) ( 808 )   Save
    References | Related Articles | Metrics
    【Objective】 The specific primers for the duplex-PCR reaction of Heterodera avenae and H. filipjevi were designed and a synchronous detection system was established for these two species. The work will provide technical supports for rapid and accurate diagnosis of cereal cyst nematodes (CCN) in wheat field of China and benefit to the integrate management strategies.【Method】The forward primers HaF8 and HfF9 respectively specific to H. avenae and H. filipjevi together with the common reverse primer HafR8 were designed according to the comparisons of mtDNA-COI sequences from 24 populations of 10 plant parasitic nematode species. The concentrations of primers and the annealing temperatures for duplex-PCR reaction were optimized and a high efficient detection system for these two CCN species was developed. The established system was applied to identify the CCN species of samples collected from wheat fields in Huanghuai region of China. 【Result】 The specific primers HaF8/HafR8 and HfF9/HafR8 obtained in this study could rapidly detect H. avenae and H. filipjevi in one PCR reaction with high specificity. The PCR product for H. avenae amplified with primers HaF8/HafR8 was 200 bp, for H. filipjevi with primers HfF9/HafR8 was 320 bp. The band sizes were easily distinguished. The optimization showed that the duplex-PCR detection system had a high specificity and amplification efficiency with the annealing temperature of primers HaF8/HfF9/HafR8 at 58. In addition, the detection system could identify H. avenae and H. filipjevi simultaneously from all detected samples with a high efficiency, when the concentration of HaF8﹕HfF9﹕HafR8 was 0.24﹕0.16﹕0.4 μmol·L-1. The sensitivities of the duplex-PCR system detection for single cyst of H. avenae and H. filipjevi were both 1/2 000 000, but for single second stage juvenile were 1/640 and 1/1 280, respectively. The duplex-PCR system was used to identify the species of 14 CCN samples collected from wheat fields in Huanghuai region of China. The results showed that the 200 bp band was amplified from 8 samples, indicating the species is H. avenae. The 320 bp band was amplified from 4 samples, indicating the species is H. filipjevi. However, both the band 200 and 320 bp were only amplified from 2 samples assuming a mixture of H. avenae and H. filipjevi occurred. The results demonstrated that the duplex-PCR detection system could be successfully applied in rapid diagnosis of the complex infection situation of CCN in fields. 【Conclusion】 The established duplex-PCR system with the specific primers designed on basis of mtDNA-COI sequences is a reliable and simple technique for simultaneously rapid detecting H. avenae and H. filipjevi in field samples on wheat.
    SOIL & FERTILIZER·WATER-SAVING IRRIGATION·AGROECOLOGY & ENVIRONMENT
    Spatial and Temporal Characteristics of Basic Soil Productivity in China
    LI Jian-jun, XU Ming-gang, XIN Jing-shu, DUAN Jian-jun, REN Yi, LI Dong-chu, HUANG Jing, SHEN Hua-ping, ZHANG Hui-min
    Scientia Agricultura Sinica. 2016, 49(8):  1510-1519.  doi:10.3864/j.issn.0578-1752.2016.08.008
    Abstract ( 480 )   HTML ( 1 )   PDF (651KB) ( 612 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this paper is to study the differences of basic soil productivity and spatial and temporal characteristics of national paddy soil in the main grain production regions of China from the Ministry of Agriculture since 1988, provide a scientific basis of fertilization for the improvement of soil fertility and rice high yield. 【Method】Combined with the actual situation of soil, and the reference model of productivity, soil quality and other correction coefficient model derived soil productivity index model (PI) and was calculated based soil fertility index model (BPI) and verification. At the same time, the national paddy soil monitoring data was used as basis, each fertility index and weight statistics resulting weighting factor were normalized by numerical normalization. PI and BPI indexes were used for comprehensive analysis, and further verified with the yield and contribution proportion of basic soil productivity phase, thereby the differences and spatial and temporal characteristics of the basic soil productivity were quantitatively analyzed. 【Result】From 1988 to 2012, both the BPI values (P<0.05) and PI values (P<0.01) showed significant increasing trends in the districts of Yangtze River, Northeast China, Southwest China, and South China. There were significant positive correlations (P<0.05) between BPI values and fertilization yields. The BPI values in the Yangtze River region increased from 0.031 to 0.108 with the largest increasing percentage of 248.4%, followed by the BPI value in the Southern China increased from 0.127 to 0.289 with an increase of 128.0%, and that in the Northeast China and Southwest China with increase of 71.7% and 65.8%, respectively, during the past 25 years. Rice yield with fertilization of each area with the enhancement of basic soil productivity of paddy showed an increasing trend. The basic soil productivity levels (BSPI value) in Yangtze River region were significantly higher than that in the other three areas. The BSPI value in the Southwest China was the lowest, while there was no significant difference between the Northeast China and the Southern China. As for regions, the trends of BSPI values coincided with the yields under the basic fertility of soil. 【Conclusion】During the past 25 years, the basic soil productivity showed an overall upward trend in the main rice production regions of China. Soil productivity could be improved with the enhancing of basic soil productivity. In addition, on the level of current farming management (from 2003 to 2012), the order of basic soil productivity among the regions were Yangtze River region>Northeast China≥Southern China>Southwest China. In the national scale, the higher the BSPI, the higher the rice yield without fertilization, the higher the contribution proportion of basic soil productivity. On the contrary, the lower the basic soil productivity, rice output is also lower, contribution proportion of basic soil productivity is also lower.
    Characteristics of Phosphorus Accumulation and Movement in a Calcareous Purple Paddy Soil Profile as Affected by Long-Term Fertilization
    FAN Hong-zhu, CHEN Qing-rui, QIN Yu-sheng, CHEN Kun, TU Shi-hua
    Scientia Agricultura Sinica. 2016, 49(8):  1520-1529.  doi:10.3864/j.issn.0578-1752.2016.08.009
    Abstract ( 403 )   HTML ( 1 )   PDF (509KB) ( 888 )   Save
    References | Related Articles | Metrics
    【Objective】 The objective of this study is to investigate the effects of long-term fertilization on phosphorus (P) accumulation and movement in profile of a calcareous purple paddy soil. 【Method】 The ongoing experiment, initiated in 1981, consisted of eight treatments including CK (no fertilizer), nitrogen fertilizer (N), nitrogen and phosphorus fertilizers (NP), nitrogen, phosphorus and potassium fertilizers (NPK), pig manure (M), pig manure plus nitrogen fertilizer (MN), pig manure plus nitrogen and phosphorus fertilizers (MNP) and pig manure plus nitrogen, phosphorus and potassium fertilizers (MNPK) with four replications. Status of P balance, accumulation and fate, soil total P (TP) and Olsen-P at 0-20 cm soil depths, and P accumulation and distribution at 0-100 cm soil depths were measured to get better understanding of the impacts of different treatments on these parameter changes after 33 years in the calcareous purple paddy soil in the Southwest China. 【Result】 Results showed that 613.12 kg·hm-2 of P in P omission treatments (CK and N alone) were absorbed by crop grain and straw after 33 years in the calcareous purple paddy soil. The P nutrient from seeds, stubble, rain and irrigation waters was about 106.61 kg·hm-2 entered into the soil. Thus soil P became deficient when P was omitted from the fertilizer program (CK and N alone), giving an average deficit value of 15.35kg·hm-2 of P nutrient. And the situation was worsened with time, changing of 0.0011 g·kg-1 in TP and 0.029 mg·kg-1 in Olsen-P annually. After 33-year different fertilizer treatments, two inorganic P treatments (NP and NPK) resulted in 1 880.03 kg·hm-2 of total P inputs and 1 275.40 kg·hm-2 of P uptake by crops. Total inputs of P were 2 532.68 kg·hm-2 and P absorbed by crop was 757.50 kg·hm-2 in two P-omitted organic manure treatments (M and MN). And P inputs and uptake were 4 305.11 and 1 436.64 kg·hm-2 in the two P-added organic manure treatments (MNP and MNPK), respectively. Thus treatments amended with P and/or organic manure yielded annual balances of total P as 18.32, 53.79 and 86.92 kg·hm-2, fate unknown P as 4.99, 34.96 and 59.39 kg·hm-2. TP changing as 0.015, 0.0018 and 0.018 g·kg-1 and Olsen-P as 1.13, 0.032 and 1.17 mg·kg-1 for the two inorganic P treatments, the two P-omitted organic manure treatments and the two P-added organic manure treatments, respectively. The P omission treatments resulted in reduction of TP with depths downward the profile, while an opposite trend was observed for Olsen-P. P treatments, however, gave P distribution patterns of highs in the top and the bottom depths but lows in the middle layers. Addition of inorganic P fertilizers could transfer P down to the 60-80 cm soil profile, while addition of organic manure alone or in combination with inorganic P fertilizers could move P further down to more than 100 cm soil depths, suggesting that organic P was more mobile than inorganic P in the soil and susceptible to loss. 【Conclusion】When soil Olsen-P reaches to an adequate level (eg. 20 mg·kg-1 as a critical value for most of cereal crops) after consecutive applications of P fertilizers for years, reduction in P rates, organic P in particular, should be considered to maintain or effectively lower soil P pool and to minimize P losses to environment.
    HORTICULTURE
    Identification and Analysis of Phosphoproteins in Red and Non-Red Apple Cultivars
    YUAN Ke-jun, CHENG Lai-liang, NIU Qing-lin, WANG Jiang-yong
    Scientia Agricultura Sinica. 2016, 49(8):  1530-1539.  doi:10.3864/j.issn.0578-1752.2016.08.010
    Abstract ( 456 )   HTML ( 2 )   PDF (406KB) ( 621 )   Save
    References | Related Articles | Metrics
    【Objective】Apple skin color is an important fruit quality factor. In order to make clear whether there is phosphorylation during apple fruit pigmentation, large scale identification of phosphoproteins and phosphopeptides was conducted. 【Method】The fruit peels of red skinned apple cultivar ‘Gala’ and non-red skinned apple cultivar ‘Golden Delicious’ were used to extract protein. Fruit protein was digested with trypsin, the resulting peptides were subjected to a step of TiO2 beads enrichment to obtain phosphopeptides, and a step of HPLC separation.Q-Exactive mass spectrometer was used to identify phosphopeptides. The software Mascot 2.2 and Proteome Discoverer1.4 (thermo) were used to identify proteins in the database Uniprot_Maloideae.fasta. Then data analysis was performed to provide information of identified phosphorylation sites in red and non-red apple cultivars.【Result】 1 323 proteins, 3 339 peptides, 225 phosphopeptides were identified and about 200-225 phosphoproteins were found in the red apple cultivars, 569 proteins, 1 152 peptides, 128 phosphopeptides were identified and about 117-128 phosphoproteins were found in the non-red apple cultivars.Comparative analysis indicated that 43 phosphopeptides were only found in non-red apple cultivars, 139 phosphopeptides were only discovered in red apple cultivars, which proved that more phosphoproteins produced in red apple cultivars, including plasma membrane H+-ATPase with two phosphorylation sites in serine, a major allergen and two protein kinases with phosphorylation site in threonine, three transcription factors, a potassium transporter and a putative protein receptor kinase. Among the identified proteins in the red and non-red apple cultivars, the phosphorylation sites were mainly found in threonine, some in serine, seldom in tyrosine. There was no phosphorylation site in the C4H, 4CL, CHS, CHI, F3′H, FLS, LDOX and UFGT proteins of anthocyanin synthesis pathway and protein WD40, the MYBR transcription factor with phosphorylation site was found in both red and non-red apple cultivars. 【Conclusion】 Some phosphoproteins only produced in red apple cultivars, including plasma membrane H+-ATPase related to apple pigmentation. Apple fruit pigmentation is possibly influenced by protein phosphorylation.
    The Analysis of Chromosome Karyotype of Malus sieversii Germplasm Resources Infraspecies in Xinjiang
    MAYNUR·turdi, ZHANG Yan-hui, QIN Wei, SI Hong-zhang, YANG Xin-feng
    Scientia Agricultura Sinica. 2016, 49(8):  1540-1549.  doi:10.3864/j.issn.0578-1752.2016.08.011
    Abstract ( 418 )   HTML ( 1 )   PDF (3354KB) ( 428 )   Save
    References | Related Articles | Metrics
    【Objective】 As a main wild apple germplasm resources in Xinjiang Ili area, as a special branch of apple resources in our country, it is of great significance to enhance the research of protecting the richness of apple germplasm resources. Aiming to discuss the genetic relationship with them, provide a scientific basis for the development and utilization of Malus sieversii resources. 【Method】 Twenty-four germplasm types which are collected and studied from Yili. Between the beginning of April and the middle of April, selecting the tender stem, tableting with the improved chromosome preforming methods for the karyotype analysis of the Xinjiang Malus sieversii. According to the chromosome number of different types and the relative length, average arm ratio, the karyotype characteristics were analyzed.【Result】 (1) All of the twenty-four type under species are diploid, whose chromosome number in all Malus sieversii types are 2n=34. And the length of the chromosome is 3.23 μm, so they belong to small chromosomes. Viewing the position of the centromere, there are different numbers of “m” and “sm” chromosomes in most of all types, the “m” type is only in very few types. Karyotype formula are four types: 2n=2x=34=34m, 2n=2x=34=30m+4sm, 2n=2x=34=28m+6sm, 2n=2x=34=32m+2sm. Viewing the length of the chromosome, except for the xiang-suan and chang-bing wild apple , the rest of the types owns long chromosome (L), middle long chromosome (M2), middle short chromosome (M1), and short chromosome (s), and it was not seen the satellite in them. (2) the range of the arm ratio is from1.27to1.56, and the symmetry the karyotype is in the range of 56.01%-63.39%, the most karyotype characteristics is 1A and 1B, 2A and 2B only exist in some few types, the percentage of which is 41.67%, 37.5%, 8.33%, 12.50%. (3) evolutionary trend diagram shows that yellow round wild apples are the most special, its karyotype asymmetry coefficient is the highest (63.39%), and also has the highest degree in its evolution, followed by Huocheng round wild apple and scent apple, and the small wild apple has  the least degree in its evolution (56.01%). The evolution trend of the rest of the 20 kinds of germplasm type has high consistency and is relatively concentrated, among them, the big flat heart wild apple's level of evolution is the lowest. (4) According to the results of cluster, 24 kinds of germplasm types are divided into three categories. The first category includes 9 kinds of germplasm types, they have higher karyotype feature similarity, most types’ average arm ratio are greater than others, combining evolutionary trend results, found their evolutionary trends are higher than other groups. The second category includes 11 kinds of germplasm types, most karyotype types are 1B, 2A, and 2B. The third category includes 4 kinds of germplasm types, their average arm ratio and karyotype asymmetry coefficient are both the minimum, karyotype type belong to 1A and 1B, all above show that the genetics of this group is relatively stable. 【Conclusion】 To some extent , there is a difference on the karyotype of xinjiang Malus sieversii,according to the karyotype, it could be classified and then judging the evolutionary trend, compared with the traditional classification method, it has revealed the genetic relationship among the germplasm types is clearer, the result of clustering can better reflect the similarities and differences among the types. Using the karyotype comparative analysis, the evolutionary trend of wild apple germplasm resources in Xinjiang Yili were revealed, which has a very important function in the further evolutionary study.
    STORAGE·FRESH-KEEPING·PROCESSING
    Effects of Sodium Carboxymethyl Cellulose on Adhesion Properties of Soybean Protein Isolate onto Porcine Bones
    ZHANG Yi-jing, CHEN Hai-juan, Lü Yi, LIU Yong-xiang, TANG Xiao-zhi, SHEN Xin-chun
    Scientia Agricultura Sinica. 2016, 49(8):  1550-1558.  doi:10.3864/j.issn.0578-1752.2016.08.012
    Abstract ( 357 )   HTML ( 1 )   PDF (1090KB) ( 540 )   Save
    References | Related Articles | Metrics
    【Objective】The effects of sodium carboxymethyl cellulose (CMC-Na) on the adhesion properties of soybean protein isolate (SPI) onto porcine bone were investigated by studying tensile bond strength, zero shear viscosity, secondary structure, hydrophobicity and scanning electronic microscopy (SEM) images. The aim of the research was to provide a basis for utilization of SPI adhesive in biomedical fields. 【Method】SPI adhesive prepared with/without CMC-Na was used to bond porcine bones in vitro and the effect of CMC-Na on tensile bond strength of SPI adhesive was measured using a tensile strength testing machine. The influence of CMC-Na on zero shear viscosity of the SPI adhesive was measured using a rheometer. The SPI adhesive with CMC-Na secondary structure was measured using circular dichroism spectra to determine the content of α-helix, β-sheet, β-turn and random coil. The influence of CMC-Na on hydrophobicity and surface morphology of SPI adhesive was measured using ANS (1-anilino-8-naphthalene sulfonate) fluorescent probe method and scanning electron microscopy (SEM), respectively. 【Result】 The tensile bond strength of the SPI adhesive increased with the increase of its concentration and reached a peak at a SPI adhesive concentration of 10%, then began to decline. The adhesive strength of 2% SPI with addition of a trace amount of CMC-Na (at 0.01%) was increased 2.4 fold (P<0.01) in comparison with the control. There was a significant positive correlation (r=0.815, P<0.05) between adhesive strength and zero shear viscosity. The addition of trace CMC-Na influenced significantly the secondary structure of the SPI adhesive. The β-sheet and β-turn content increased from 42.2% to 49.1% and from 2.1% to 7.3%, respectively, while the α-helix and random coil content declined from 28.0% to 19.7% and from 27.7% to 23.9%, respectively. The hydrophobicity of the SPI adhesive with CMC-Na was increased significantly; even moderate hydrophobic modification of the SPI adhesive with CMC-Na benefited the increase of its adhesive strength. SEM showed that particles on the surface of the SPI adhesive with addition of CMC-Na arrayed more uniformly and compact than the control, which was beneficial for its adhering to the bones. 【Conclusion】The addition of a trace amount of CMC-Na caused significant changes in the secondary structure of the SPI adhesive, the decline in α-helix content and an increase in β-sheet content, all of which indicated the unfolding of protein molecules and the aggregation of unfolding protein molecules. The increase in the hydrophobicity and zero shear viscosity of the SPI adhesive resulted in the significant increase of bond strength of low concentration SPI adhesive onto porcine bone. Since low concentration SPI might be easier to be absorbed by bodies than at a high concentration, our results after the addition of CMC-Na suggest that SPI adhesive has a great potential to be utilized as an adhesive in medical fields.
    The Impact of Alkaline Hydrogen Peroxide Pretreatment on the Chemical Composition and Enzymatic Hydrolyzability of Apple Pomace
    YOU Yi-na, DENG Hong, MENG Yong-hong, FANG Jie, GUO Yu-rong, ZHANG Ying
    Scientia Agricultura Sinica. 2016, 49(8):  1559-1566.  doi:10.3864/j.issn.0578-1752.2016.08.013
    Abstract ( 421 )   HTML ( 1 )   PDF (1629KB) ( 548 )   Save
    References | Related Articles | Metrics
    【Objective】 The aim of this study was to research the impact of a 4.5% alkaline hydrogen peroxide (AHP) pretreatment at pH 11.5 on the chemical composition of apple pomace, the lignin removal rate, and the enzymatic hydrolyzability of cellulase. 【Method】 The temperature and time of pretreatment were optimized in accordance with the index of the lignin removal rate in this study. After filtering to collect solid components and drying, the AHP pretreated apple pomace was obtained. According to the content of cellulose and hemicellulose, the content of lignin, and total sugar content of enzymatic hydrolysis of apple pomace before AHP, the effects of different pretreated time on the content and recovery of cellulose and hemicellulose, the content and removal rate of lignin, and the enzymatic hydrolyzability were analyzed under the optimal pretreatment temperature. Utilizing a scanning electron microscope (SEM), thermogravimetry/differential thermogravimetry (TG/DTG), and fourier transform infrared (FTIR) characterized the physical and chemical composition change of apple pomace before and after AHP. 【Result】 The temperature and time of pretreatment had significant effects on the lignin removal rate. Considering various factors, especially the economic benefits, when the time was 2 hours and the temperature was 50℃, the lignin removal rate was optimal with 56.68%. The composition variation and enzymatic hydrolyzability of apple pomace after different pretreated times was analyzed under optimal pretreated temperature. When pretreated time was 2 hours, the recovery of cellulose and hemicellulose was 99.86%, which was very close to the content of untreated apple pomace; the enzymatic hydrolyzability was 0.54 g·g-1, which was 200% of untreated apple pomace. The SEM images showed that the physical structure was multi-hole and loose, the fiber bundle became loose and rough, and the internal surface area was indefinitely increased after apple pomace treated with AHP. TG/DTG demonstrated that AHP could apparently increase the purity of cellulose monomer and reduce the content of lignin and residue. The FTIR study revealed that AHP could substantially disrupt the chemical structure of lignin, and the structure of guaiacyl, syringyl, and p-hydroxylphengl of lignin was destroyed. Thus, the lignin content of apple pomace was reduced after AHP. 【Conclusion】 This paper illustrated that AHP was an efficient pretreatment agent of apple pomace, and that the effect of pretreatment was closely associated with temperature and time.
    ANIMAL SCIENCE·VETERINARY SCIENCERE·SOURCE INSECT
    Generation of Beta-Mannanase Transgenic Clone Pig and Analysis
    ZHANG Mao, ZHANG Guan-guan, LIU De-wu, CAI Geng-yuan, WU Zhen-fang, LI Zi-cong
    Scientia Agricultura Sinica. 2016, 49(8):  1567-1576.  doi:10.3864/j.issn.0578-1752.2016.08.014
    Abstract ( 319 )   HTML ( 3 )   PDF (1437KB) ( 475 )   Save
    References | Related Articles | Metrics
    【Objective】The objective of this study is to transfect the plasmid pPSPBGP-manA into porcine fetal fibroblast cells and screening of transgenic cell lines with G418. Transgenic pigs were generated by somatic cell nuclear transfer and identified by PCR and Southern blot analyses, beta-mannose mRNA protein were identified by RT-PCR and Western blot, so as to provide a scientific basis for the further study of the genetically modified pig.【Method】The plasmid pPSPBGP-manA was linearized with Not I and purified. The Duroc pig fetal fibroblast cells were cultured to the third generation and using liposome method to transfer plasmid into porcine fetal fibroblasts. Then the transgenic cells were selected by G418 and the transgenic pigs were obtained by somatic cell nuclear transfer. Genomic DNA was isolated from the new born porcine tail tissue by phenol-chloroform extraction. One microgram of each genomic DNA sample was used as the template for a single PCR and Southern blot analyses. Total RNA was isolated from frozen parotid gland, sublingual gland, mandibular gland, brain, heart, liver, spleen, lung, kidney and stomach tissues of transgenic pigs using Triol method. RT-PCR of pig total RNA was performed to identify the expression of manA gene in different tissues. The expression level of manA gene in different individuals and tissues was detected by relative quantitative PCR. The saliva beta-mannose activity analysis with DNS method and the content of nutrients in feces was determined. Moreover, the protein expression of exogenous manA gene in transgenic pig was detected by Western blotting. 【Result】 Stable transfection of the transgenic cell line was obtained by G418 screening and PCR identification. Twenty-one cloning pigs were produced by somatic cell nuclear transfer, PCR and Southern blotting showed that there were 16 transgenic pigs and the positive rate was 76%.Transgenic pig salivary beta-mannosidase enzyme activity was (0.092±0.003)U·mL-1 and fecal crude protein content decreased significantly. RT-PCR and relative quantitative PCR analyses demonstrated that manA is strongly expressed in parotid gland and sublingual gland, but is not present in mandibular gland, brain, heart, liver, spleen, lung, kidney or stomach, and the manA expression level in parotid gland was higher than in sublingual gland. The beta-mannanase protein was detected in the parotid and sublingual gland by Western blotting. 【Conclusion】 The transgenic cell line and transgenic pig were successfully obtained and the exogenous manA gene could be specific ally expressed in parotid and sublingual gland.
    The Interaction of Colonization of Akkermansia muciniphila in Gastrointestinal Tract and Its Host
    FENG Ze-meng, BAO Xian-ying, YIN Yu-long
    Scientia Agricultura Sinica. 2016, 49(8):  1577-1584.  doi:10.3864/j.issn.0578-1752.2016.08.015
    Abstract ( 481 )   HTML ( 10 )   PDF (371KB) ( 973 )   Save
    References | Related Articles | Metrics
    The gastrointestinal tract is a big harbour for various microbes, which have a great influence on the metabolism and development of the host. These intestinal microbiota can be divided into microbiota in inner lumen and in mucus layer, and both have important roles in nutrients delivery and prevention against pathogenic microorganism invasion. Akkermansia muciniphila, a kind of gram-negative bacteria, specially degrades mucin, has a growth preference in animal mucus and has a broad effect on the host. In the gastrointestinal tract, the flora of Akkermansia muciniphila degrade mucin and oligosaccharides, produce short chain fatty acid and propionic acid, respectively, which provide energy to the host and also promote their colonization. At the same time, the degradation of mucin will lead to more mucin secretion, thereby lowering the host protein deposition. The colonization of Akkermansia muciniphila reduce fat deposition, delay the formation of diabetes. Akkermansia muciniphila have no serious pathogenicity, and suitable abundance will promote the development of the host immune system and intestinal health. With the development of more scientific research, the importance of intestinal microbiota will be the hot topic. However, the mechanism of the action of Akkermansia muciniphila to the host is still unclear. The colonized environment, physiological characteristics, host nutrition metabolism disturbance, relation with metabolic diseases, and host immune regulation of Akkermansia muciniphila were summarized in the present paper. Akkermansia muciniphila is a good potential biomarker, and can be applied in nutritional status, metabolic diseases, immunity and even cancer detection, and worth to be further studied.
    Isolation Identification and Bioinformatics of Differences Protein in Plasma of Cows Suffer from Fatty Liver with SELDI-TOF-MS Techniques
    XU Chuang, ZHU Kui-ling, CHEN Yuan-yuan, YANG Wei, XIA Cheng, ZHANG Hong-you, WU Ling, SHU Shi, SHEN Tai-yu, YU Hong-jiang, XU Qiu-shi, ZHANG Zi-yang
    Scientia Agricultura Sinica. 2016, 49(8):  1585-1598.  doi:10.3864/j.issn.0578-1752.2016.08.016
    Abstract ( 385 )   HTML ( 1 )   PDF (1892KB) ( 434 )   Save
    References | Related Articles | Metrics
    Objective】 Fatty liver is an important metabolic disorder of dairy cows in transition period, and 50% of postpartum cows are affected. Productivity redution, poor reproductive performance, weak immune function, failure of liver function and premature death are related to fatty liver in postpartum cows. Surface-enhanced laser desorption/ionization (SELDI) time-of-fight (TOF) mass spectrometry (MS) is a sensitive proteomics technology. The aim of this study was to determine the differences between the plasma proteomic profiles of health cows and those with fatty liver. 【Method】  Forty Holstein cows at 7-28 d postpartum with 1-2 parties were selected from an intensive dairy farm in Heilongjiang province. Ten milliliters of blood was taken from the jugular vein of the 40 cows, the blood was placed in an anticoagulant tube and were mixed with the anticoagulant evenly. Blood samples were centrifuged at 3000r/min for 5 min, the serum were stored at −80°C. According to the concentration of serum glycerin trilaurate (TG>0.20mmol·L-1) and beta-hydroxybutyric acid (BHBA>0.97mmol·L-1), 20 cows were assigned to the fatty liver group (T) and 20 to the control group (C). Plasma proteins mass spectra were tested bysurface-enhanced laser desorption/ionization time-of-flight mass spectrometry (SELDI-TOF-MS). The peak figures of the two groups were analyzed using the Ciphergen ProteinChip Software. The peak data of the two groups were analyzed using wilcoxon rank sum test, p values were calculated to confirm the difference of the two groups. Different peaks were selected if P<0.01. m/z values of different peaks were compared with theoretical m/z values using Swissport database. The analogous proteins were identified as a prediction result. 【Result】The 39 significantly different peptide peaks which were identified in the T group were compared with the theoretical m/z values in Swissport database. A total of 26 predictable differences peaks were obtained, 11 kinds of proteins were predicted. Results showed that the levels of these 11 proteins were reduced in T group compared with C group. The 11 kinds of protein networks figure were analyzed by bioinformatics analysis (Network,GO,Pathway) and cattle genetic network search, networks analysis results were got. The two differential protein amyloid precursor protein (APP) and fibrinogen alpha chain (FGA) were found in database, relative Network analysis results were obtained. The BinGO plug-ins in Cytoscape software were used to analyze the 11 kinds of protein and GO analysis results were got. There were 9 kinds of different proteins were found by the software search, including FGA, serum amyloid A protein (SAA), plasma protease c1 inhibitor (SERPING1 or C1INH), apolipoprotein c-Ⅲ (Apo-CⅢ), hepcidin (HAMP), osteopontin (OPN or SPP1), transthyretin (TTR), Cystatin-c (CysC, CST), neurosecretory protein (VGA). There were 6 kinds of different proteins were searched in the KEGG pathway database, including FGA, SERPING1, Apo-CⅢ, APP, CysC and SPP1. They may related to the pathogenesis of fatty liver of dairy cows. 【Conclusion】The differential expression protein in serum between health cows and fatty liver cows were separated effectively by SELDI-TOF-MS technology, which may play an important role in the process of metabolism of liver and development of fatty liver. Therefore, this study has important theoretical values to reveal new mechanism of fatty liver and its effect on bio-function of dairy cows. Further studies need to be conducted to reveal the impact of different proteins on the pathogenesis of fatty liver in dairy cows.
    Cloning and Expression of cAMP Response Element Binding Protein (CREB) Gene of Apis cerana cerana
    ZHANG Li-zhen, WANG Zi-long, ZENG Zhi-jiang
    Scientia Agricultura Sinica. 2016, 49(8):  1599-1606.  doi:10.3864/j.issn.0578-1752.2016.08.017
    Abstract ( 385 )   HTML ( 1 )   PDF (4674KB) ( 641 )   Save
    References | Related Articles | Metrics
    【Objective】 The objective of this study is to clone the full-length cDNA sequence of the cAMP response element binding protein gene of Apis cerana cerana (AcCREB), predict the gene and its physicochemical properties of the corresponding encoded protein, and then clarify its expression in the brain tissues, which will provide a research basis of the physiological function of AcCREB in the learning and memory. 【Method】Total RNA was extracted from the dissected brain tissues of A. cerana cerana (A.c.c.) as the main experimental materials. On this basis, the AcCREB was cloned using RT-PCR technique. The forward and reverse sequences were then spliced by Bioedit to obtain the full-length cDNA using the Seqman of DNAstar software, and its amino acid sequences were translated using Bioedit software according to the six-box translation. The sequences was analyzed using BLASTn and BLASTp. Multiple sequence alignment and homology were analyzed using ClustalX. A phylogenetic tree was constructed using neighbor-joining method of software MEGA. The conserved structural domains were deduced by analyzing the SMART database. In addition, the paraffin sections of brain were made based on the dissected brain tissues, and the distribution of its expression in the brain was investigated by in situ hybridization histochemistry. 【Result】The full length of AcCREB cDNA is 890 bp, which encodes 240 amino acids. The sequence was submitted to GenBank (Accession number KC814690). The molecular weight and an isoelectric point of AcCREB protein were 25.691 kD and 5.82, respectively. Structural domain analysis indicated that the encoded protein contained two highly conserved structures of AA 89-131 PKID and AA 176-237 BRLA. Homology comparison illustrated that AcCREB has a similarity with CREB of Apis mellifera (AmCREB), Bombus impatiens (BiCREB), Megachile rotundata (MrCREB), Bombus terrestris (BtCREB), Harpegnathos saltator (HsCREB), Bombyx mori (BmCREB), Aedes aegypt (AaCREB),Anopheles gambiae (AgCREB), Homo sapiens (HsCREB)and Mus musculus (MmCREB), with the similarity of 98.76%, 98.35%, 97.11%, 94.09%, 72.22%, 58.97%, 44.69%, 41.69%, 36.89%, and 36.89%, respectively. Meanwhile, the phylogenetic tree analysis showed that AcCREB was first clustered with AmCREB, BiCREB, BtCREB. Moreover, in situ hybridization showed that AcCREB staining was mainly localized to the Kenyon cells of the mushroom bodies, cells around the antennal lobes, cells between medulla and lobula, and display asymmetrical expression between the right and left brain of worker bees in A.c.c..【Conclusion】The amino sequences of AcCREB own high homology with AmCREB, BiCREB, MrCREB, BtCREB, and the distribution of the gene in the brains of honeybee suggested that AcCREB may be involved in the process of learning and memory of the A. c. c. which provides useful information for future studies of CREB in the leaning-memory of honeybees.
    Identification and Expression Analysis of BmYki-1 in the Silkworm (Bombyx mori)
    YANG Li-qun, JIA Le-mei, TANG Mei, CHEN Yi-biao, CUI Hong-juan
    Scientia Agricultura Sinica. 2016, 49(8):  1607-1616.  doi:10.3864/j.issn.0578-1752.2016.08.018
    Abstract ( 422 )   HTML ( 3 )   PDF (1613KB) ( 538 )   Save
    References | Related Articles | Metrics
    【Objective】Yki, a transcriptional coactivator, is phosphorylated and activated by upstream kinases cascade of the Hippo signaling pathway, and then regulates the downstream genes expression to control organ size, proliferation and apoptosis of cell. As a model of Lepidopteran insect, little is known about the Hippo signaling pathway involved in the silkworm (Bombyx mori). The objective of this study is to identify B. mori BmYki-1 and analyze its sequence information and express characteristics, laying the foundation for further study of Hippo signaling pathways in midgut, which will help us better understand Hippo signaling pathways in intestinal regulatory mechanism of steady maintaining and tissue damage repairing.【Method】Based on mammals and drosophila Yki protein sequence, B. mori homologous Yki gene sequences was identified using NCBI databases and silkworm genome database. B. mori Dazao strain being research material, full-length sequence of two Yki spliced variants were cloned by PCR and RACE. According to bioinformatics methods, gene sequences and protein structure characteristics of the two spliced variants were analyzed. Here, function of BmYki-1 in B. mori intestinal tissue was focused. The spatial-temporal expression profile of BmYki-1 was investigated by semi-quantitative RT-PCR. BmYki-1 was constructed into a transit expression vector, and transfected into B. mori embryonic cell line to analyze the subcellular location of BmYki-1. qRT-PCR was utilized to detect the expression of BmYki-1 in B. mori intestinal after treated with bleomycin.【Result】There are two alternatively spliced forms of BmYki, namely BmYki-1 and BmYki-2, whose Gene ID are BGIBMGA0082-TA and BGIBMGA0081-TA in silkworm genome database. The ORF of the two spliceosomes consist of 1 329 and 1 227 bp, encoding 442 and 408 amino acids, with 48 and 44.1 kD protein molecular weight and 5.18 and 5.50 isoelectric point, respectively. In addition, both spliceosomes have two WW domains from 139th to 171th and 220th to 252th amino acids. A multi species phylogenetic tree demonstrated that BmYki-1 was most closely related to Amyelois transitella, which is Lepidoptera insect like B. mori, and was comparatively closely related to Coleoptera and Hymenoptera insects. However, BmYki-1 had a distant relationship with that of Diptera insect, and furthest with vertebrates. Expression profile showed that BmYki-1 was specifically expressed in the midgut and silk gland, and the expression was significantly up-regulated from 2nd molting stage in the midgut and reached the top at early-to-mid stage of 5th instar. Overexpressed BmYki-1 in BmE cells was mainly distributed in the nucleus. BmYki-1 was significantly up-regulated in the midgut treated with bleomycin compared to the control.【Conclusion】The BmYki-1 cDNA was cloned and its expression patterns in B. mori were analyzed. The subcellular location of BmYki-1 was investigated and mainly located in nucleus. BmYki-1 may play a critical role in the development and repair of damaged midgut tissue of B. mori.
    RESEARCH NOTES
    Quantifying the Effect of the Addition Amount of Clay on the Compressive Characteristics of Different Organic Carbon Contents in Black Soil
    CHAI Xin, AN Jing, ZHANG Yu-long, ZOU Hong-tao, YU Na, FAN Qing-feng
    Scientia Agricultura Sinica. 2016, 49(8):  1617-1624.  doi:10.3864/j.issn.0578-1752.2016.08.019
    Abstract ( 345 )   HTML ( 1 )   PDF (365KB) ( 326 )   Save
    References | Related Articles | Metrics
    【Objective】The compaction of cultivated soils by agricultural machines considerably affects both the structure and physical properties of soil, and thus can have major impacts on crop production and the environment. The objective of this study is to explore the effect of the addition amount of clay on the compressive characteristics of different organic carbon contents in black soil.【Method】Different amounts of clay (0, 10%, 20%, 40% denoted as C0, C1, C2 and C3) were added to three different amounts of total organic carbon (TOC) of black soil (M0, M1, M2) in order to prepare soil samples (M0C0, M0C1, M0C2, M0C3, M1C0, M1C1, M1C2, M1C3, M2C0, M2C1, M2C2, M2C3). The soil moisture content was adjusted to 20% and bulk density 1.1 g·cm-³. Then performing uniaxial compression tests were made indoor and the pre-compression stress values (σp) and compression index (Cc) were determined.【Result】The higher the TOC was, the higher the slope of the plastic part of the compression curve, when soils were at the same clay condition, with the increase of TOC, soil void increased when the same pressure was applied. The amount of added clay, TOC and the interaction between them all had a significant effect on both σp and Cc (P<0.05). The amount of added clay and TOC both had a significant positive correlation with σp of black soil, and the amount of added clay had no significant positive correlation with the Cc, while the correlation between TOC and Cc was not obvious.【Conclusion】With the increase of the amount of organic carbon content and clay, pre-compression stress of black soil increased. The more organic carbon content and clay is, the higher Cc will be, the easier the deformation will be after compression.
    Effects of Low Oxygen Stress on the Root Function and Enzyme Activities Related to Nitrogen Metabolism in Roots of Rice Seedlings
    XU Chun-mei, CHEN Li-ping, WANG Dan-ying, CHEN Song, ZHANG Xiu-fu, SHI Qing-hua
    Scientia Agricultura Sinica. 2016, 49(8):  1625-1634.  doi:10.3864/j.issn.0578-1752.2016.08.020
    Abstract ( 475 )   HTML ( 6 )   PDF (452KB) ( 705 )   Save
    References | Related Articles | Metrics
    【Objective】 This study was conducted to research the effects of low oxygen stress on root growth, respiratory function and enzyme activities related to nitrogen metabolism of rice seedlings, explore the physiological mechanism of rice root system, tillers and biomass accumulation, root respiratory intensity and the enzyme activities related to nitrogen metabolism under short low oxygen stress, in order to lay a foundation for finding out the role of low oxygen stress injury and protective mechanism of rice seedlings.【Method】The formal experiment was conducted at China National Rice Research Institute in 2015. Two rice cultivars Xiushui 09 and Chunyou 84 were grown in hydroponic condition at the space 20 cm× 20 cm. At seven days after transplanting with online dissolved oxygen meter (oxygen nitrogen regulator) set two dissolved oxygen contents (DOTs ) levels such as low oxygen(DOT 0 to 1.0 mg·L-1)and control ( conventional hydroponics, without oxygen regulation). The portable dissolved oxygen meters (HACHHQ30d, American) was used to examine the oxygen concentration in the nutrient solution. The changes of root activity, respiratory intensity, enzyme activities related to nitrogen metabolism were determined every 3 days after low oxygen stress treatment. Plant high, tillers, chlorophyll content, root system and dry matters and other indexes were determined and analyzed at the end of treatment.【Result】Rice seedlings growth was inhibited under short low oxygen stress. At the end of low oxygen stress treatment, when compared to the CK, the tillering numbers and dry matter accumulation of Xiushui 09 decreased by 20.0% and 7.78%, respectively, and those of Chunyou84 decreased by15.38% and 6.28%, respectively. The amount of coarse roots (average diameter>0.8 mm), fine root (average diameter between 0.05 and 0.1 mm) and root volume were less than the control too; chlorophyll content, root nitrogen content and soluble protein content of root were inhibited under low oxygen stress. The amount of all indexes of Xiushui 09 were decreased by 6.67%, 15.11% and 10.86%, respectively, and those of Chunyou 84 were decreased by 5.18%, 13.25% and 6.67%, respectively, which illustrated that Xiushui 09 was more sensitive to low oxygen stress. Higher plant were observed under low oxygen stress, Xiushu 09 increased by 7.98% and Chunyou 84 by 3.30%, respectively, compared with CK. The effect of low oxygen stress on rice seedling root respiratory intensity was inhibited-promoted-inhibited, with the prolongation of the treatment time, the effect of inhibition was reduced. The change trend of root activity was consistent with the root respiratory intensity. The activity of nitrate reductase (NR) of rice root was promoted under low oxygen stress. The effect of glutamine synthase (GS) of rice was promoted from 0 to 6 days and was inhibited from 9 to 12 days under low oxygen stress. Low oxygen stress affect the activity of glutamate dehydrogenase (NADH-GDH) was relevant to the rice variety, it was promoted-inhibited of Xiushui 09, and it was inhibited-promoted-inhibited of Chunyou 84. Short low oxygen stress inhibited the growth of rice seedlings.【Conclusion】It was concluded that rice seedlings are probably by improving respiration consumption and changing the metabolic pathway to alleviate low oxygen stress injury, and to maintain their survival under the condition of low oxygen stress.