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    Crop Genetics · Breeding · Germplasm Resources
    Experimental and genomic evidence for the indica-type cytoplasmic effect in Oryza sativa L. ssp. japonica
    LIU You-hong, TANG Liang, XU Quan, MA Dian-rong, ZHAO Ming-hui, SUN Jian, CHEN Wen-fu
    2016, 15(10): 2183-2191.  DOI: 10.1016/S2095-3119(15)61190-X
    Abstract ( )   PDF in ScienceDirect  
        Cytoplasmic effects are important agronomical phenomena that have generated widespread interest in both theory and application. In the present study, five high yield rice cultivars (Oryza sativa L. ssp. japonica) in large-scale cultivation in northeast China were determined to possess Oryza sativa L. ssp. indica-type cytoplasm using cytoplasmic subspecies-specific molecular markers. This was confirmed by cytoplasmic genome-wide single nucleotide polymorphisms (SNPs) and functional gene sequencing. Two of these five japonica cultivars were core breeding parents with high yield and the other three were super-high-yield varieties registered by the Ministry of Agriculture of China. We constructed nuclear substitution lines to further demonstrate whether and how this indica-type cytoplasm contributed to yield improvement by comparing yield components. The results showed that under the same japonica nuclear background, the lines with indica-type cytoplasm had a significant decrease in tillers in exchange for increased grain number per panicle compared with their recurrent parents. Our results implied that botanical basis of this cytoplasmic effect was to reduce the plant’s branching differentiation to produce more floral organs under the constant nutrition. Our findings open another door for the utilization of inter-subspecific hybridization for the improvement of rice cultivar.
    Association mapping of quantitative trait loci for yield-related agronomic traits in rice (Oryza sativa L.)
    XU Fei-fei, JIN Liang, HUANG Yan, TONG Chuan, CHEN Ya-ling, BAO Jin-song
    2016, 15(10): 2192-2202.  DOI: 10.1016/S2095-3119(15)61244-8
    Abstract ( )   PDF in ScienceDirect  
        High yield in rice mainly depends on large grain weight, ideal plant architecture and proper flowering time adapting to various geographic regions. To help achieve higher yield, phenotype variations of heading date (HD), plant architecture and grain shape in a panel of 416 rice accessions were investigated in this study. A total of 143 markers including 100 simple sequence repeat (SSR) markers and 43 gene-tagged markers were employed in association mapping to detect quantitative trait loci (QTL) responsible for these variations. Among the 7 subpopulations, POP5 in japonica group showed the largest values of HD and grain width (GW), but the smallest values of grain length (GL) and grain length to width ratio (GLW). Among the six indica groups, POP7 had the largest values of HD, GL, GLW, and 1 000-grain weight (TGW). A total of 27 QTLs were detected underlying these phenotypic variations in single year, while 12 of them could be detected in 2006 and 2007. GS3 marker was closely associated with GL, GW and GLW, and widely distributed in different groups. The starch synthesis related gene markers, SSI, SSIIa, SBE1, AGPL4, and ISA1, were linked to plant height (PH), panicle length (PL), flag leaf length (FLL), GW, and GLW. The SSR markers, RM267, RM340 and RM346, were linked to at least two traits. Therefore, these new markers will probably be used to improve rice grain yield or plant architecture when performing marker-assisted selection of proper alleles.
    Characterization of A- and B-type starch granules in Chinese wheat cultivars
    ZHANG Yan, GUO Qi, FENG Nan, WANG Jin-rong, WANG Shu-jun, HE Zhong-hu
    2016, 15(10): 2203-2214.  DOI: 10.1016/S2095-3119(15)61305-3
    Abstract ( )   PDF in ScienceDirect  
        Starch is the major component of wheat flour and serves as a multifunctional ingredient in food industry. The objective of the present study was to investigate starch granule size distribution of Chinese wheat cultivars, and to compare structure and functionality of starches in four leading cultivars Zhongmai 175, CA12092, Lunxuan 987, and Zhongyou 206. A wide variation in volume percentages of A- and B-type starch granules among genotypes was observed. Volume percentages of A- and B-type granules had ranges of 68.4–88.9% and 9.7–27.9% in the first cropping seasons, 74.1–90.1% and 7.2–25.3% in the second. Wheat cultivars with higher volume percentages of A- and B-type granules could serve as parents in breeding program for selecting high and low amylose wheat cultivars, respectively. In comparison with the B-type starch granules, the A-type granules starch showed difference in three aspects: (1) higher amount of ordered short-range structure and a lower relative crystallinity, (2) higher gelatinization onset (To) temperatures and enthalpies (ΔH), and lower gelatinization conclusion temperatures (Tc), (3) greater peak, though, and final viscosity, and lower breakdown viscosity and pasting temperature. It provides important information for breeders to develop potentially useful cultivars with particular functional properties of their starches suited to specific applications.
    Functional analysis of a wheat pleiotropic drug resistance gene involved in Fusarium head blight resistance
    WANG Gui-ping, HOU Wen-qian, ZHANG Lei, WU Hong-yan, ZHAO Lan-fei, DU Xu-ye, MA Xin, LI An-fei, WANG Hong-wei, KONG Ling-rang
    2016, 15(10): 2215-2227.  DOI: 10.1016/S2095-3119(16)61362-X
    Abstract ( )   PDF in ScienceDirect  
        The pleiotropic drug resistance (PDR) sub-family of adenosine triphosphate (ATP)-binding cassette (ABC) transporter had been reported to participate in diverse biological processes of plant. In this study, we cloned three novel PDR genes in Fusarium head blight (FHB) resistant wheat cultivar Ning 7840, which were located on wheat chromosomes 6A, 6B and 6D. In phylogeny, these genes were members of cluster I together with AePDR7 and BdPDR7. Subcellular localization analysis showed that TaPDR7 was expressed on the plasmalemma. The quantitative real time PCR (RT-PCR) analysis showed that this gene and its probable orthologues in chromosomes 6B and 6D were both up-regulated sharply at 48 h after infected by Fusarium graminearum and trichothecene deoxynivalenol (DON) in spike. When knocking down the transcripts of all TaPDR7 members by barely stripe mosaic virus-induced gene silencing (BSMV-VIGS) system, it could promote the F. graminearum hyphae growth and made larger pathogen inoculation points in Ning 7840, which suggested that TaPDR7 might play an important role in response to F. graminearum. Although salicylic acid (SA), methyl jasmonate (MeJA) and abscisic acid (ABA) had been reported to possibly regulate wheat FHB resistance, here, we found that the three members of TaPDR7 were negatively regulated by these three hormones but positively regulated by indoleacetic acid (IAA).
    Characterization of dwarf mutants and molecular mapping of a dwarf locus in soybean
    CHENG Wen, GAO Jin-shan, FENG Xing-xing, SHAO Qun, YANG Su-xin, FENG Xian-zhong
    2016, 15(10): 2228-2236.  DOI: 10.1016/S2095-3119(15)61312-0
    Abstract ( )   PDF in ScienceDirect  
        Plant height is one of the most important traits in soybean. The semi-dwarf soybean cultivars could improve the ability of lodging resistance to obtain higher yield. To broaden the dwarfism germplasm resources in soybean, 44 dwarf mutants were identified from a gamma rays mutagenized M2 population. Two of these mutants, Gmdwf1 (Glycine max dwarf 1) and Gmdwf2 (Glycine max dwarf 2), were investigated in this study. Genetic analysis showed that both mutants were inherited in a recessive manner and their mutated regions were delimited to a 2.610-Mb region on chromosome 1 by preliminary mapping. Further fine mapping study proved that the two mutants had a common deletion region of 1.552 Mb in the target region, which was located in a novel locus site without being reported previously. The dwarfism of Gmdwf1 could not be rescued by gibberellin (GA) and brassinolide (BR) treatments, which indicated that the biosynthesis of these hormones was not deficient in Gmdwf1.
    Genetic variation and population structure of the mushroom Pleurotus ferulae in China inferred from nuclear DNA analysis
    ZHAO Meng-ran, HUANG Chen-yang, WU Xiang-li, CHEN Qiang, QU Ji-bin, LI Yan-chun, GAO Wei, ZHANG Jin-xia
    2016, 15(10): 2237-2246.  DOI: 10.1016/S2095-3119(16)61383-7
    Abstract ( )   PDF in ScienceDirect  
        To investigate the genetic diversity of an edible fungus Pleurotus ferulae, a total of 89 wild samples collected from six geographical locations in the Xinjiang Uygur Autonomous Region of China and two geographical locations in Italy, were analyzed using three DNA fragments including the translation elongation factor (EF1α), the second largest subunit of the RNA polymerase II (RPB2) and the largest subunit of the RNA polymerase II (RPB1). The results indicated relatively abundant genetic variability in the wild resources of P. ferulae. The analysis of molecular variance (AMOVA) showed that the vast majority of the genetic variation was found within geographical populations. Both the Chinese populations and the Italian populations of P. ferulae displayed a limited genetic differentiation. The degree of differentiation between the Chinese populations and the Italian populations was obviously higher than that between the populations from the same region, and moreover the genetic differentiation among all the tested populations was correlated to the geographical distance. The phylogeny analyses confirmed that samples from China and Italy belonged to another genetic group separated from Pleurotus eryngii. They were closely related to each other but were clustered according to their geographical origins, which implied the Chinese populations were highly differentiated from the Italian populations because of distance isolation, and the two populations from different regions might be still in the process of allopatric divergence.
    Contribution of ear photosynthesis to grain yield under rainfed and irrigation conditions for winter wheat cultivars released in the past 30 years in North China Plain
    WANG Yun-qi, XI Wen-xing, WANG Zhi-min, WANG Bin, XU Xue-xin, HAN Mei-kun, ZHOU Shun-li, ZHANG Ying-hua
    2016, 15(10): 2247-2256.  DOI: 10.1016/S2095-3119(16)61408-9
    Abstract ( )  
        To understand the contribution of ear photosynthesis to grain yield and its response to water supply in the improvement of winter wheat, 15 cultivars released from 1980 to 2012 in North China Plain (NCP) were planted under rainfed and irrigated conditions from 2011 to 2013, and the ear photosynthesis was tested by ear shading. During the past 30 years, grain yield significantly increased, the flag leaf area slightly increased under irrigated condition but decreased significantly under rainfed condition, the ratio of grain weight:leaf area significantly increased, and the contribution of ear photosynthesis to grain yield changed from 33.6 to 64.5% and from 32.2 to 57.2% under rainfed and irrigated conditions, respectively. Grain yield, yield components, and ratio of grain weight:leaf area were positively related with contribution of ear photosynthesis. The increase in grain yield in winter wheat was related with improvement in ear photosynthesis contribution in NCP, especially under rainfed condition.
    Establishment of ANEDr model for evaluating absorbed-nitrogen effects on wheat dry matter production
    ZHAO Jiao, TAO Hong-bin, LIAO Shu-hua, WANG Pu
    2016, 15(10): 2257-2265.  DOI: 10.1016/S2095-3119(16)61352-7
    Abstract ( )   PDF in ScienceDirect  
        Applying mathematic models to evaluate absorbed-N effects on dry matter production at different developmental stages would help determine proper nitrogen management according to crop demands and yield target. Two field trials were carried out for establishing absorbed-N effects on dry matter production (ANEDr) model, using uniform design in 2010–2011 and 2012–2013 winter wheat growing seasons in Hebei Province, China. Another field trial was carried out in 2010–2011 for model validation. Dry matter and N concentration in leaf and non-leaf organs were measured at setting, jointing, anthesis, and maturity. Theory of best linear unbiased prediction (BLUP) was applied to analyse the N effects of leaf and non-leaf organs on dry matter production. Within ANEDr model, four N-affected phases at each stage were concerned, leaf absorbed-N effect before this stage, non-leaf organ absorbed-N effect before this stage, leaf absorbed-N effect at this stage, and non-leaf organ absorbed-N effect at this stage. In addition, developmental processes, genotype characters and temperature were three factors that determine each N effect. It was demonstrated that ANEDr model can precisely quantify absorbed-N effects on dry matter production with high correlation coefficient (r=0.95). Comparing with other models, ANEDr model considered both leaf and non-leaf organs according to developmental processes of winter wheat, showed higher flexibility and simplicity, thus could be applied to different environments, cultivars and crops after parameter adjustment.
    Proteomic analysis of salt and osmotic-drought stress in alfalfa seedlings
    MA Qiao-li, KANG Jun-mei, LONG Rui-cai, CUI Yan-jun, ZHANG Tie-jun, XIONG Jun-bo, YANG Qingchuan, SUN Yan
    2016, 15(10): 2266-2278.  DOI: 10.1016/S2095-3119(15)61280-1
    Abstract ( )   PDF in ScienceDirect  
        Alfalfa is widely grown and is one of the most important forage crops in the world, but its growth and biomass production are markedly reduced under salt and drought stress, particularly during the early seedling stage. To identify the mechanisms behind salt and drought responsiveness at the alfalfa seedling stage, the proteins expressed were analyzed under no-treatment, 200 mol L–1 NaCl and 180 g L–1 PEG treatment conditions during the seedling stage. Out of more than 800 protein spots detected on two-dimensional electrophoresis (2-DE) gels, 35 proteins showed statistically significant responses (P<0.05) to NaCl and PEG stress, which were selected for tandem mass spectrometric identification, owing to their good resolution and abundance levels, and 32 proteins were positively identified. The identified proteins were divided into seven functional categories: photosynthetic metabolism, protein biosynthesis, folding and assembly, carbohydrate metabolism-associated proteins, stress defense related protein, metabolism of nucleic acid, other function categories and unknown proteins. Our results suggested that these proteins may play roles in alfalfa adaptation to salt and drought stress. Further study of these proteins will provide insights into the molecular mechanisms of abiotic stress and the discovery of new candidate markers in alfalfa.
    Overexpression of vacuolar proton pump ATPase (V-H+-ATPase) subunits B, C and H confers tolerance to salt and saline-alkali stresses in transgenic alfalfa (Medicago sativa L.)
    WANG Fa-wei, WANG Chao, SUN Yao, WANG Nan, LI Xiao-wei, DONG Yuan-yuan, Yao Na, Liu Xiu-ming, CHEN Huan, CHEN Xi-feng, WANG Zhen-min, LI Hai-yan
    2016, 15(10): 2279-2289.  DOI: 10.1016/S2095-3119(16)61399-0
    Abstract ( )   PDF in ScienceDirect  
        The vacuolar proton pump ATPase (V-H+-ATPase), which is a multi-subunit membrane protein complex, plays a major role in the activation of ion and nutrient transport and has been suggested to be involved in several physiological processes, such as cell expansion and salt tolerance. In this study, three genes encoding V-H+-ATPase subunits B (ScVHA-B, GenBank: JF826506), C (ScVHA-C, GenBank: JF826507) and H (ScVHA-H, GenBank: JF826508) were isolated from the halophyte Suaeda corniculata. The transcript levels of ScVHA-B, ScVHA-C and ScVHA-H were increased by salt, drought and saline-alkali treatments. V-H+-ATPase activity was also examined under salt, drought and saline-alkali stresses. The results showed that V-H+-ATPase activity was correlated with salt, drought and saline-alkali stress. Furthermore, V-H+-ATPase subunits B, C and H (ScVHA-B, ScVHA-C and ScVHA-H) from S. corniculata were introduced separately into the alfalfa genome. The transgenic alfalfa was verified by Southern and Northern blot analysis. During salt and saline-alkali stresses, transgenic linevacuolar proton pump, salt tolerance, saline-alkali tolerance, alfalfa
    s carrying the B, C and H subunits had higher germination rates than the wild type (WT). More free proline, higher superoxide dismutase (SOD) activity and lower malondialdehyde (MDA) levels were detected in the transgenic plants under salt and saline-alkali treatments. Moreover, the ScVHA-B transgenic lines showed greater tolerance to salt and saline-alkali stresses than the WT. These results suggest that overexpression of ScVHA-B, ScVHA-C and ScVHA-H improves tolerance to salt and saline-alkali stresses in transgenic alfalfa.
    Plant Protection
    Pyramiding of Pi46 and Pita to improve blast resistance and to evaluate the resistance effect of the two R genes
    XIAO Wu-ming, LUO Li-xin, WANG Hui, GUO Tao, LIU Yong-zhu, ZHOU Ji-yong, ZHU Xiao-yuan, YANG Qi-yun, CHEN Zhi-qiang
    2016, 15(10): 2290-2298.  DOI: 10.1016/S2095-3119(16)61415-6
    Abstract ( )   PDF in ScienceDirect  
        Utilization of R (resistance) genes to develop resistant cultivars is an effective strategy to combat against rice blast disease. In this study, R genes Pi46 and Pita in a resistant accession H4 were introgressed into an elite restorer line Hang-Hui-179 (HH179) using the marker-assisted backcross breeding (MABB) procedure. As a result, three improved lines (e.g., R1791 carrying Pi46 alone, R1792 carrying Pita alone and R1793 carrying both Pi46 and Pita) were developed. The three improved lines had significant genetic similarities with the recurrent parent HH179. Thus, they and HH179 could be recognized as near isogenic lines (NILs). The resistance spectrum of the three improved lines, which was tested at seedling stage, reached 91.1, 64.7 and 97.1%, respectively. This was markedly broader than that of HH179 (23.5%). Interestingly, R1793 showed resistance to panicle blast but neither R1791 nor R1792 exhibited resistance at two natural blast nurseries. The results implied that the stacking of Pi46 and Pita resulted in enhanced resistance, which was unachievable by either R gene alone. Further comparison indicated that the three improved lines were similar to HH179 in multiple agronomic traits; including plant height, tillers per plant, panicle length, spikelet fertility, and 1 000-grain weight. Thus, the three improved lines with different R genes can be used as new sources of resistance for developing variety. There is a complementary effect between the two R genes Pi46 and Pita.
    Genetic diversity of Ustilago hordei in Tibetan areas as revealed by RAPD and SSR
    ZHOU Yu, CHAO Gui-mei, LIU Jia-jia, ZHU Ming-qi, WANG Yang, FENG Bai-li
    2016, 15(10): 2299-2308.  DOI: 10.1016/S2095-3119(16)61413-2
    Abstract ( )   PDF in ScienceDirect  
        Covered smut, which is caused by Ustilago hordei (Pers.) Lagerh., is one of the most damaging diseases of highland barley (Hordeum vulgare Linn. var. nudum Hook. f) in Tibetan areas of China. To understand the molecular diversity of U. hordei, a total of 27 isolates, which were collected from highland barley plants from Tibet, Sichuan, Qinghai, and Gansu provinces/autonomous region, were analyzed using random amplified polymorphic DNA (RAPD) and simple sequence repeat (SSR) markers. Among the 100 RAPD primers used, 24 primers exhibited polymorphism. A total of 111 fragments were amplified, of which 103 were polymorphic with a polymorphic rate of 92.79%. The average observed number of alleles (Na), effective number of alleles (Ne), Nei’s genetic diversity (H), Shannon’s information index (I) and polymorphism information content (PIC) value in the RAPD markers were 1.9279, 1.5016, 0.2974, 0.4503 and 0.6428, respectively. For the SSR markers, 40 of the 111 primer pairs exhibited polymorphism and provided a total of 119 bands, of which 109 were polymorphic and accounted for 91.60% of the total bands. The Na, Ne, H, I and PIC values of the SSR markers were 1.9160, 1.4639, 0.2757, 0.4211 and 0.4340, respectively. The similarity coefficients ranged from 0.4957 to 0.9261 with an average of 0.7028 among all the 27 isolates used. The dendrogram, which was developed based on the RAPD and SSR combined marker dataset showed that the 27 U. hordei isolates were divided into 3 clusters at similarity coefficient of 0.7314. We determined that RAPD and SSR markers can be successfully used to assess the genetic variation among U. hordei isolates. The RAPD markers revealed higher levels of genetic polymorphism than did the SSR markers in this study. There existed a moderate genetic difference among isolates. The molecular variation and differentiation was somewhat associated with geographical origin but not for all of the isolates.
    Effect of heat shock on the susceptibility of Frankliniella occidentalis (Thysanoptera: Thripidae) to insecticides
    ZHANG Bin, ZUO Tai-qiang, LI Hong-gang, SUN Li-juan, WANG Si-fang, ZHENG Chang-ying, WAN Fang-hao
    2016, 15(10): 2309-2318.  DOI: 10.1016/S2095-3119(16)61431-4
    Abstract ( )   PDF in ScienceDirect  
        Currently, insecticides are considered as the primary approach for controlling western flower thrips, Frankliniella occidentalis (Pergande) (Thysanoptera: Thripidae). However, the heavy use of insecticides resulted in high insect resistance and serious environmental pollution. Given its characteristics of ease of operation and environmental friendliness, insect control using high temperature is receiving considerable renewed research interest. However, although the combination of insecticides and high temperature to control F. occidentalis has been studied before, few studies have focused on the short-term effect of such treatment. In a laboratory study, F. occidentalis adults and second-instar nymphs were exposed to 45°C for 2 h. Then, their susceptibility to acetamiprid, spinosad, methomyl, and beta-cypermethrin was tested after different periods of recovery time (2–36 h). Additionally, the specific activity of three detoxification enzymes (esterase, glutathione S-transferase, and cytochrome p450 (CYP) monooxygenase) of the treated insects was determined. The results indicated that the fluctuation of susceptibility to insecticides and detoxification enzyme activity during F. occidentalis recovery from heat shock are related. Furthermore, several recovery time points (2, 30, and 36 h) of significant susceptibility to four tested insecticides compared with the control were found during the treatment of adults that were heat-shocked. Recovery time points of higher susceptibility compared with the control depended on different insecticides during the second-instar nymph recovery from heat shock. Interestingly, the fluctuation of CYP monooxygenase activity exhibited a trend that was similar to the fluctuation of susceptibility to insecticides (especially spinosad) during the recovery from heat shock of adults. In addition, the glutathione S-transferase and CYP monooxygenase activity trend was similar to the trend of susceptibility to spinosad during the recovery from heat shock of second-instar nymphs. Our results provide a new approach for controlling F. occidentalis using the combined heat shock and insecticide. This effectively enhances the control efficiency of heat shock and significantly reduces the application of insecticides
    Immunotoxicity assessment of cadinene sesquiterpenes from Eupatorium adenophorum in mice
    OUYANG Can-bin, LIU Xiao-man, YAN Dong-dong, LI Yuan, WANG Qiu-xia, CAO Ao-cheng, GUO Mei-xia
    2016, 15(10): 2319-2325.  DOI: 10.1016/S2095-3119(16)61403-X
    Abstract ( )   PDF in ScienceDirect  
        Sesquiterpenes in Eupatorium adenophorum are abundant in leaves and have great development potential as biopesticides. The toxicity of sesquiterpenes in immune cells and their corresponding immune functions are not fully understood. We evaluated the immunotoxicity of two cadinene sesquiterpenes 2-deoxo-2-(acetyloxy)-9-oxoageraphorone (DAOA) and 9-oxo-10,11-dehydro-agerophorone (ODA) by using histopathology and toxicology methods in vitro and in vivo in lymphocytes and natural killer cells in Kunming mice. The mice were given single doses of 75, 150 and 300 mg kg−1 body weight (BW) of DAOA/ODA every day for a week. Serious damage to the thymus and spleen was found in tissue images with clear lysis reduction numbers and a loosened arrangement of splenocytes and thymocytes to the mice treated with 150–300 mg kg−1 DAOA/ODA. Mice cytology was also affected with significant cellular alterations, increased splenocytes apoptosis rates (P<0.01), proliferation reduction (P<0.05) and natural killer cells activities reduction (P<0.05) when given 150–300 mg kg−1 DAOA/ODA, the severities of which were dose-dependent. However, a 75 mg kg−1 dose of DAOA/ODA showed no change in tissue or cytology after the 7 day treatment, and therefore was considered to be within acceptable safety parameters. Taken together, cadinene sesquiterpenes, as a type of toxic botanical component, have low environmental risks in small doses and should be further studied for their use as biopesticides.
    Animal Science · Veterinary Science
    Sperm pretreatment with glutathione improves IVF embryos development through increasing the viability and antioxidative capacity of sex-sorted and unsorted bull semen
    HU Ting-xi, ZHU Hua-bin, SUN Wei-jun, HAO Hai-sheng, ZHAO Xue-ming, DU Wei-hua, WANG Zong-li
    2016, 15(10): 2326-2335.  DOI: 10.1016/S2095-3119(16)61402-8
    Abstract ( )  
        The antioxidant of reduced glutathione (GSH) is the most abundant thiol in cells for the maintenance of the intracellular redox balance. The study aimed to assay the effect of sperm treatment with GSH before incubation with oocytes on the development potential of embryos obtained by in vitro fertilization (IVF). Also the mitochondrial membrane potential (ΔΨm), plasma membrane integrity (viability), DNA fragmentation, reactive oxygen species (ROS) content, superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) activities, methane dicarboxylic aldehyde (MDA) level as indices of lipid peroxidation in sex-sorted and unsorted sperm from three bulls were investigated using flow cytometry and enzyme-labeled instrument individually. The results showed that 2 mmol L–1 GSH increased significantly the cleavage rate (86.68% vs. 82.78%), 4- to 8-cell rate (82.30% vs. 73.43%) and blastocyst rate (43.15% vs. 35.24%) of IVF embryos compared with untreated group. Furthermore, addition of GSH increased significantly the ΔΨm and viability, decreased the ratio of DNA fragmentation in sex-sorted or unsorted semen (P<0.05), except the sex-sorted semen from bull 019. Similarly, activities of SOD, CAT and GPx were increased significantly. However, the contents of MDA were decreased significantly both in sex-sorted and unsorted semen treated with GSH (P<0.05). These results suggest that sperm pretreatment with GSH during IVF can maintain better the viability and fertility of sperm through reducing apoptosis and increasing the antioxidant capacity, which improves the IVF embryos development.
    Effects of sustained cold and heat stress on energy intake, growth and mitochondrial function of broiler chickens
    ZHANG Shao-shuai, SU Hong-guang, ZHOU Ying, LI Xiu-mei, FENG Jing-hai, ZHANG Min-hong
    2016, 15(10): 2336-2342.  DOI: 10.1016/S2095-3119(15)61314-4
    Abstract ( )   PDF in ScienceDirect  
        To study the correlation of broiler chickens with energy intake, growth and mitochondrial function which exposed to sustained cold and heat stress and to find out the comfortable temperature, 288 broiler chickens (21-day with (748±26) g, 144 males and 144 females) were divided randomly into six temperature-controlled chambers. Each chamber contained six cages including eight AA broilers per cage, each cage as a repeat. After acclimation for one week (temperature, 21°C; relative humidity, 60%), the temperature of each chamber was adjusted (finished within 1 h) respectively to 10, 14, 18, 22, 26, or 30°C (RH, 60%) for a 14-day experimental period. After treatment, gross energy intake (GEI), metabolizable energy intake (MEI), the ratio of MEI/BW, metabolizability, average daily gain (ADG), the concentration of liver mitochondria protein and cytochrome c oxidase (CCO) were measured respectively. Our results confirmed that when the temperature over 26°C for 14 days, GEI, MEI and CCO activities were decreased significantly (P<0.05), but the concentration of liver mitochondria protein was increased and metabolizability of broilers was not influenced (P>0.05). Compared with treatment for 14 days, the ratio of MEI/body weight (BW) were also decreased when the temperature over 26°C after temperature stress for 7 days (P<0.05), meanwhile mitochondrial protein concentration was increased at 10°C and CCO activity was not affected (P>0.05). Additionally at 22°C, the ADG reached the maximal value. When kept in uncomfortable temperatures for a long time, the ADG and CCO activities of broiler were reduced, which was accompanied by mitochondrial hyperplasia. In summary, our study focused on the performance of broilers during sustained cold and heat environmental temperatures ranging from 10 to 30°C. From the point of view of energy utilization, moreover, 22 to 26°C is comfortable for 28–42 days broilers. And these could provide the theoretical basis on the high efficient production.
    Animal Science · Veterinary Science
    Initial detection of the quorum sensing autoinducer activity in the rumen of goats in vivo and in vitro
    RAN Tao, ZHOU Chuan-she, XU Li-wei, GENG Mei-mei, TAN Zhi-liang, TANG Shao-xun, WANG Min, HAN Xue-feng, KANG Jin-he
    2016, 15(10): 2343-2352.  DOI: 10.1016/S2095-3119(16)61417-X
    Abstract ( )   PDF in ScienceDirect  
        Quorum sensing (QS) is a type of microbe-microbe communication system that is widespread among the microbial world, particularly among microorganisms that are symbiotic with plants and animals. Thereby, the cell-cell signalling is likely to occur in an anaerobic rumen environment, which is a complex microbial ecosystem. In this study, using six ruminally fistulated Liuyang black goats as experimental animals, we aimed to detect the activity of quorum sensing autoinducers (AI) both in vivo and in vitro and to clone the luxS gene that encoded autoinducer-2 (AI-2) synthase of microbial samples that were collected from the rumen of goats. Neutral detergent fiber (NDF) and soluble starch were the two types of substrates that were used for in vitro fermentation. The fermented fluid samples were collected at 0, 2, 4, 6, 8, 12, 24, 36, and 48 h of incubation. The acyl-homoserine lactones (AHLs) activity was determined using gas chromatography-mass spectrometer (GC-MS) analysis. However, none of the rumen fluid extracts that were collected from the goat rumen showed the same or similar fragmentation pattern to AHLs standards. Meanwhile, the AI-2 activity, assayed using a Vibrio harveyi BB170 bioassay, was negative in all samples that were collected from the goat rumen and from in vitro fermentation fluids. Our results indicated that the activities of AHLs and AI-2 were not detected in the ruminal contents from six goats and in ruminal fluids obtained from in vitro fermentation at different sampling time-points. However, the homologues of luxS in Prevotella ruminicola were cloned from in vivo and in vitro ruminal fluids. We concluded that AHLs and AI-2 could not be detected in in vivo and in vitro ruminal fluids of goats using the current detection techniques under current dietary conditions. However, the microbes that inhabited the goat rumen had the potential ability to secrete AI-2 signaling molecules and to communicate with each other via AI-2-mediated QS because of the presence of luxS.
    Selection and characterisation of lactic acid bacteria isolated from different origins for ensiling Robinia pseudoacacia and Morus alba L. leaves
    NI Kui-kui, YANG Hui-xiao, HUA Wei, WANG Yan-ping, PANG Hui-li
    2016, 15(10): 2353-2362.  DOI: 10.1016/S2095-3119(15)61251-5
    Abstract ( )   PDF in ScienceDirect  
        The objective of this study was to isolate lactic acid bacteria (LAB) strains from different origins and to select the best strains for ensiling Robinia pseudoacacia (RB) and Morus alba L. (MB) leaves. The LAB strains were inoculated into the extracted liquid obtained from RB and MB leaves to evaluate the fermentation products. 11 LAB strains were selected for further experiments based on the highest products of lactic or acetic acid, including 1 strain of Weissella confusa, 2 of Lactobacillus reuteri and 8 of Lactobacillus plantarum. The API 50 CH fermentation experiment indicated that all of the selected 11 LAB strains utilised most of the carbohydrates. All the strains grew at temperatures between 10 and 45°C and at a pH of 3.5 to 4.5; however, L. reuteri F7 and F8 tolerated a pH as low as 3.0. All 11 LAB strains showed antibacterial activity against Listeria monocytogens, Escherichia coil, Salmonella sp. and Acetobacter pasteurianus; however, after excluding the effect of organic acids, only F7 and F8 still exhibited antibacterial activity. The present study indicated that the selected 11 LAB strains could be used to prepare silages of RB and MB leaves, especially L. reuteri F7 and F8.
    Development and optimization of a double antibody sandwich ELISA for the detection of goose T cell surface CD8α molecule
    ZHANG Wei, CHENG Bei-bei, CHEN Shun, WANG Ming-shu, JIA Ren-yong, ZHU De-kang, LIU Mafeng, LIU Fei, SUN Kun-feng, YANG Qiao, WU Ying, CHEN Xiao-yue, CHENG An-chun
    2016, 15(10): 2363-2368.  DOI: 10.1016/S2095-3119(16)61345-X
    Abstract ( )   PDF in ScienceDirect  
        CD8, a glycoprotein on the surface of T cells, is involved in the defense against viral infection and plays significant roles in antigen presentation and in the antiviral immune response. CD8 is composed of two chains. Of these, the CD8α chain was chosen for the detection because it involved in both the CD8αα homodimer and the CD8αβ heterodimer. Here, we established a double antibody sandwich enzyme-linked immunosorbent assay (DAS-ELISA) for specific detection of goose CD8α (goCD8α). The results showed that the optimal coated antibody and antigen dilutions were 1:50 (the antibody titer was 1:12 800) and 1:32 (0.3 ng mL–1), respectively, while the optimal capture antibody and horseradish peroxidase (HRP)-labelled goat anti-rabbit IgG dilutions were 1:50 (the antibody titer was 1:51 200) and 1:4 000 (the antibody titer was 1:5 000), respectively. The optimal blocking buffer was 5% bovine serum albumin (BSA). The best incubating condition was overnight at 4°C, the best blocking time was 120 min and the best anti-capture antibody working time was 150 min. In addition, the minimum dose detectable by DAS-ELISA was 5×10–3 ng mL–1. Most importantly, goCD8α expression levels in goose spleen mononuclear cells (MNCs) post-Goose parvoviruse (GPV) infection were found to be significantly up-regulated using the DAS-ELISA method, which was consistent with previous results obtained using real-time quantitative PCR. In conclusion, the DAS-ELISA method reported here is a novel, specific technique for the clinical detection of goCD8α.
    Soil & Fertilization﹒Irrigation﹒Plant Nutrition﹒ Agro-Ecology & Environment
    Screening of a microbial consortium with efficient corn stover degradation ability at low temperature
    Qinggeer , GAO Ju-lin, YU Xiao-fang, ZHANG Bao-lin, WANG Zhi-gang, Borjigin Naoganchaolu, HU Shu-ping, SUN Ji-ying, XIE Min, WANG Zhen
    2016, 15(10): 2369-2379.  DOI: 10.1016/S2095-3119(15)61272-2
    Abstract ( )  
        To speed up the degradation of corn stover directly returned to soil at low temperature, the corn stover-degrading microbial consortium GF-20, acclimated to biological decomposition in the frigid region, was successfully constructed under a long-term limiting substrate. To evaluate its potential in accelerating the decomposition of un-pretreated corn stover, the decomposing property, fermentation dynamic and the microbial diversity were analyzed. GF-20 degraded corn stover by 32% after 15-day fermentation at 10°C. Peak activities of filter paperlyase (FPA), β-glucosidases (CB), endoglucanases (Cx), and cellobiohydrolases (C1) were 1.15, 1.67, 1.73, and 1.42 U mL–1, appearing at the 6th, 3rd, 11th, and 9th d, respectively. The pH averaged at 6.73–8.42, and the optical density (OD) value peaked at 1.87 at the 120 h of the degradation process. Cellulase, hemicellulase and lignin in corn stover were persistently degraded by 44.85, 43.85 and 25.29% at the end of incubation. Result of denaturing gradient gel electrophoresis (DGGE) profiles demonstrated that GF-20 had a stable component structure under switching the temperature and pH. The composition of the GF-20 was also analyzed by constructing bacterial 16S rDNA clone library and fungal 18SrDNA-PCR-DGGE. Twenty-two bacterial clones and four fungal bands were detected and identified dominant bacteria represented by Cellvibrio mixtus subsp., Azospira oryzae, Arcobacter defluyii, and Clostridium populeti and the fungi were mainly identified as related to Trichosporon sp.
    Field performance of alternate wetting and drying furrow irrigation on tomato crop growth, yield, water use efficiency, quality and profitability
    Khokan Kumer Sarker, M. A. R. Akanda, S. K. Biswas, D. K. Roy, A. Khatun, M. A. Goffar
    2016, 15(10): 2380-2392.  DOI: 10.1016/S2095-3119(16)61370-9
    Abstract ( )  
        Sustainable irrigation method is now essential for adaptation and adoption in the areas where water resources are limited. Therefore, a field experiment was conducted to test the performance of alternate wetting and drying furrow irrigation (AWDFI) on crop growth, yield, water use efficiency (WUE), fruit quality and profitability analysis of tomato. The experiment was laid out in randomized complete block design with six treatments replicated thrice during the dry seasons of 2013-2014 and 2014-2015. Irrigation water was applied through three ways of furrow: AWDFI, fixed wetting and drying furrow irrigation (FWDFI) and traditional (every) furrow irrigation (TFI). Each irrigation method was divided into two levels: irrigation up to 100 and 80% field capacity (FC). Results showed that plant biomass (dry matter) and marketable fruit yield of tomato did not differ significantly between the treatments of AWDFI and TFI, but significant difference was observed in AWDFI and in TFI compared to FWDFI at same irrigation level. AWDFI saved irrigation water by 35 to 38% for the irrigation levels up to 80 and 100% FC, compared to the TFI, respectively. AWDFI improved WUE by around 37 to 40% compared to TFI when irrigated with 100 and 80% FC, respectively. Fruit quality (total soluble solids and pulp) was found greater in AWDFI than in TFI. Net return from AWDFI technique was found nearly similar compared to TFI and more than FWDFI. The benefit cost ratio was viewed higher in AWDFI than in TFI and FWDFI by 2.8, 8.7 and 11, 10.4% when irrigation water was applied up to 100 and 80% FC, respectively. Unit production cost was obtained lower in AWDFI compared to TFI and FWDFI. However, AWDFI is a useful water-saving furrow irrigation technique which may resolve as an alternative choice compared with TFI in the areas where available water and supply methods are limited to irrigation.
    Prediction model for mercury transfer from soil to corn grain and its cross-species extrapolation
    HU Hai-yan, LI Zhao-jun, FENG Yao, LIU Yuan-wang, XUE Jian-ming, Murray Davis, LIANG Yong-chao
    2016, 15(10): 2393-2402.  DOI: 10.1016/S2095-3119(15)61261-8
    Abstract ( )   PDF in ScienceDirect  
        In this study the transfer characteristics of mercury (Hg) from a wide range of Chinese soils to corn grain (cultivar Zhengdan 958) were investigated. Prediction models were developed for determining the Hg bioconcentration factor (BCF) of Zhengdan 958 from soil, including the soil properties, such as pH, organic matter (OM) concentration, cation exchange capacity (CEC), total nitrogen concentration (TN), total phosphorus concentration (TP), total potassium concentration (TK), and total Hg concentration (THg), using multiple stepwise regression analysis. These prediction models were applied to other non-model corn cultivars using a cross-species extrapolation approach. The results indicated that the soil pH was the most important factor associated with the transfer of Hg from soil to corn grain. Hg bioaccumulation in corn grain increased with the decreasing pH. No significant differences were found between two prediction models derived from different rates of Hg applied to the soil as HgCl2. The prediction models established in this study can be applied to other non-model corn cultivars and are useful for predicting Hg bioconcentration in corn grain and assessing the ecological risk of Hg in different soils.
    Monitoring of winter wheat distribution and phenological phases based on MODIS time-series: A case study in the Yellow River Delta, China
    CHU Lin, LIU Qing-sheng, HUANG Chong, LIU Gao-huan
    2016, 15(10): 2403-2416.  DOI: 10.1016/S2095-3119(15)61319-3
    Abstract ( )   PDF in ScienceDirect  
        Accurate winter wheat identification and phenology extraction are essential for field management and agricultural policy making. Here, we present mechanisms of winter wheat discrimination and phenological detection in the Yellow River Delta (YRD) region using moderate resolution imaging spectroradiometer (MODIS) time-series data. The normalized difference vegetation index (NDVI) was obtained by calculating the surface reflectance in red and infrared. We used the Savitzky-Golay filter to smooth time series NDVI curves. We adopted a two-step classification to identify winter wheat. The first step was designed to mask out non-vegetation classes, and the second step aimed to identify winter wheat from other vegetation based on its phenological features. We used the double Gaussian model and the maximum curvature method to extract phenology. Due to the characteristics of the time-series profiles for winter wheat, a double Gaussian function method was selected to fit the temporal profile. A maximum curvature method was performed to extract phenological phases. Phenological phases such as the green-up, heading and harvesting phases were detected when the NDVI curvature exhibited local maximum values. The extracted phenological dates then were validated with records of the ground observations. The spatial patterns of phenological phases were investigated. This study concluded that, for winter wheat, the accuracy of classification is 87.07%, and the accuracy of planting acreage is 90.09%. The phenological result was comparable to the ground observation at the municipal level. The average green-up date for the whole region occurred on March 5, the average heading date occurred on May 9, and the average harvesting date occurred on June 5. The spatial distribution of the phenology for winter wheat showed a significant gradual delay from the southwest to the northeast. This study demonstrates the effectiveness of our proposed method for winter wheat classification and phenology detection.
    Linking a farmer crop selection model (FCS) with an agronomic model (EPIC) to simulate cropping pattern in Northeast China
    HE Ying-bin, CAI Wei-min
    2016, 15(10): 2417-2425.  DOI: 10.1016/S2095-3119(15)61247-3
    Abstract ( )   PDF in ScienceDirect  
      In this paper, authors established a farmer crop selection model (FCS) for the three provinces of Liaoning, Jilin and Heilongjiang of the Northeast China. With linking to the environmental policy integrated climate model (EPIC), the simulated results of FCS model for maize, rice and soybean were spatialized with 1 km×1 km grids to obtain cropping pattern. The reference map of spatial distribution for the three staple crops acquired by remote sensing imageries was applied to validate the simulated cropping pattern. The results showed that (1) the total simulation accuracy for the study area was 78.62%, which proved simulation method was applicable and feasible; (2) simulation accuracy for Jilin Province was the highest among the three provinces with a rate of 82.45% since its simple cropping system and not complex topography; (3) simulation accuracy for maize was the best among the three staple crops with a ratio of 81.14% because the study area is very suitable for maize growth. We hope this study could provide the reference for cropping pattern forecasting and decision-making.
    Emergy analysis of paddy farming in Hunan Province, China: A new perspective on sustainable development of agriculture
    YI Ting, XIANG Ping-an
    2016, 15(10): 2426-2436.  DOI: 10.1016/S2095-3119(16)61375-8
    Abstract ( )   PDF in ScienceDirect  
        The multi-functionality of paddy farming has become a hot issue recently. Paddy farming provides numerous ecosystem services that are crucial to human well-being. However, evaluation of the contribution of paddy farming to human well-being usually focus on its economic value, while its non-market services are usually ignored. Only evaluating the market profits or market relative benefits cannot reflect comprehensively the contribution of paddy farming to people’s well-being. This will affect people’s choices for or against paddy farming activities and people’s opt for invest or not invest in it. A comprehensive evaluation of paddy farming can provide an important reference for the government and society to conserve the multi-functionality of paddy farming and achieve sustainable development. To this end, this paper reports a case evaluation of paddy farming in Hunan, the largest rice producing as well as rice yield province in China, and uses emergy theory to make a comprehensive evaluation for paddy farming. The emergy evaluation results of the paddy ecosystem in Hunan are as follows: in 2010, the input emergy of the paddy ecosystem in Hunan is 2.51E+22 sej and the output emergy is 6.31E+22 sej. For the input emergy, the part from natural resources is 1.96E+21 sej and the part from human society is 2.32E+22 sej; for the output emergy, the part from products is 2.22E+22 sej, the part from impositive externality is 4.16E+22 sej and the part from negative externality is –7.41E+20 sej. Taking the non-market outputs into consideration, the gains from the human economic society’s 1 $ input in paddy farming, emergy sustainability index (ESI) and emergy profit rate are respectively 2.73 $, 3.53 and 151.31%. If the evaluation leave out the non-market output, the three indexes are only 0.96 $, 1.24 and 30.67%. The research results show that non-market services of paddy farming contribute significantly to human well-being. Therefore, in order to protect the multi-functionality of paddy farming and achieve the sustainable management, the government should take reasonable measures and make incentive plans.