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2017 Vol. 16 No. 09 Previous Issue    Next Issue

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Review
Crop Science
Horticulture
Plant Protection
Animal Science · Veterinary Medicine
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Review

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Rice molecular markers and genetic mapping: Current status and prospects Ghulam Shabir, Kashif Aslam, Abdul Rehman Khan, Muhammad Shahid, Hamid Manzoor, Sibgha Noreen, Mueen Alam Khan, Muhammad Baber, Muhammad Sabar, Shahid Masood Shah, Muhammad Arif 2017, 16(09): 1879-1891.  DOI: 10.1016/S2095-3119(16)61591-5 Abstract ( )   PDF in ScienceDirect       Dramatic changes in climatic conditions that supplement the biotic and abiotic stresses pose severe threat to the sustainable rice production and have made it a difficult task for rice molecular breeders to enhance production and productivity under these stress factors. The main focus of rice molecular breeders is to understand the fundamentals of molecular pathways involved in complex agronomic traits to increase the yield. The availability of complete rice genome sequence and recent improvements in rice genomics research has made it possible to detect and map accurately a large number of genes by using linkage to DNA markers. Linkage mapping is an effective approach to identify the genetic markers which are co-segregating with target traits within the family. The ideas of genetic diversity, quantitative trait locus (QTL) mapping, and marker-assisted selection (MAS) are evolving into more efficient concepts of linkage disequilibrium (LD) also called association mapping and genomic selection (GS), respectively. The use of cost-effective DNA markers derived from the fine mapped position of the genes for important agronomic traits will provide opportunities for breeders to develop high-yielding, stress-resistant, and better quality rice cultivars. Here we focus on the progress of molecular marker technologies, their application in genetic mapping and evolution of association mapping techniques in rice.

Crop Science

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Characterization and expression analysis of a novel RING-HC gene, ZmRHCP1, involved in brace root development and abiotic stress responses in maize LI Wen-lan, SUN Qi, LI Wen-cai, YU Yan-li, ZHAO Meng, MENG Zhao-dong 2017, 16(09): 1892-1899.  DOI: 10.1016/S2095-3119(16)61576-9 Abstract ( )   PDF in ScienceDirect      RING is a really interesting new gene which plays important regulatory roles in many developmental processes as well as in plant-environment interactions. In the present report, the ZmRHCP1 gene encoding a putative RING-HC protein was isolated from maize and characterized. The ZmRHCP1 protein contained 310 amino acid residues with a conserved RING-HC zinc-finger motif and two transmembrane (TM) domains. ZmRHCP1 was expressed ubiquitously in various organs (root, stem, leaf, seedling, immature ear, and tassel), but its transcript levels were higher in vegetative organs than in reproductive organs. Moreover, the expression pattern of ZmRHCP1 in brace roots indicated that ZmRHCP1 functions in brace root initiation. In addition, ZmRHCP1 expression was regulated by abiotic stresses. The expression results suggested that ZmRHCP1 plays important roles in brace root development and abiotic stress responses. The findings of the present study provide important information to help us understand the function of ZmRHCP1 in maize.

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QTL effects and epistatic interaction for flowering time and branch number in a soybean mapping population of Japanese×Chinese cultivars YANG Guang, ZHAI Hong, WU Hong-yan, ZHANG Xing-zheng, Lü Shi-xiang, WANG Ya-ying, LI Yu-qiu, HU Bo, WANG Lu, WEN Zi-xiang, WANG De-chun, WANG Shao-dong, Kyuya Harada, XIA Zheng-jun, XIE Fu-ti 2017, 16(09): 1900-1912.  DOI: 10.1016/S2095-3119(16)61539-3 Abstract ( )   PDF in ScienceDirect      Flowering time and branching type are important agronomic traits related to the adaptability and yield of soybean. Molecular bases for major flowering time or maturity loci, E1 to E4, have been identified. However, more flowering time genes in cultivars with different genetic backgrounds are needed to be mapped and cloned for a better understanding of flowering time regulation in soybean. In this study, we developed a population of Japanese cultivar (Toyomusume)×Chinese cultivar (Suinong 10) to map novel quantitative trait locus (QTL) for flowering time and branch number. A genetic linkage map of a F2 population was constructed using 1 306 polymorphic single nucleotide polymorphism (SNP) markers using Illumina SoySNP8k iSelect BeadChip containing 7 189 (SNPs). Two major QTLs at E1 and E9, and two minor QTLs at a novel locus, qFT2_1 and at E3 region were mapped. Using other sets of F2 populations and their derived progenies, the existence of a novel QTL of qFT2_1 was verified. qBR6_1, the major QTL for branch number was mapped to the proximate to the E1 gene, inferring that E1 gene or neighboring genetic factor is significantly contributing to the branch number.

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A callus transformation system for gene functional studies in soybean XU Kun, ZHANG Xiao-mei, FAN Cheng-ming, CHEN Fu-lu, ZHU Jin-long, ZHANG Shi-long, CHEN Qing-shan, FU Yong-fu 2017, 16(09): 1913-1922.  DOI: 10.1016/S2095-3119(16)61621-0 Abstract ( )   PDF in ScienceDirect       Obtaining transgenic plants is a common method for analyzing gene function. Unfortunately, stable genetic transformation is difficult to achieve, especially for plants (e.g., soybean), which are recalcitrant to genetic transformation. Transient expression systems, such as Arabidopsis protoplast, Nicotiana leaves, and onion bulb leaves are widely used for gene functional studies. A simple method for obtaining transgenic soybean callus tissues was reported recently. We extend this system with simplified culture conditions to gene functional studies, including promoter analysis, expression and subcellular localization of the target protein, and protein-protein interaction. We also evaluate the plasticity of this system with soybean varieties, different vector constructs, and various Agrobacterium strains. The results indicated that the callus transformation system is efficient and adaptable for gene functional investigation in soybean genotype-, vector-, and Agrobacterium strain-independent modes. We demonstrated an easy set-up and practical homologous strategy for soybean gene functional studies.

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Temperature and solar radiation utilization of rice for yield formation with different mechanized planting methods in the lower reaches of the Yangtze River, China XING Zhi-peng, WU Pei, ZHU Ming, QIAN Hai-jun, HU Ya-jie, GUO Bao-wei, WEI Hai-yan, XU Ke, HUO Zhong-yang, DAI Qi-gen, ZHANG Hong-cheng 2017, 16(09): 1923-1935.  DOI: 10.1016/S2095-3119(16)61596-4 Abstract ( )   PDF in ScienceDirect   Several studies have demonstrated the effect of planting methods on rice yield, but information on the climate resources is limited.  This study aims to reveal the effects of planting methods on climate resources associated with rice yield in a rice-wheat rotation system in the lower reaches of the Yangtze River, China.  Field experiments were conducted in 2014 and 2015 with two japonica, two indica hybrid, and two japonica-indica hybrid varieties grown under three mechanized planting methods: carpet seedling of mechanical transplanting (CT), mechanical direct seeding (DS), and pot-hole seedling of mechanical transplanting (PT).  The rice yield and total This study was financially supported by grants from the Major Independent Innovation Project in Jiangsu Province, China (CX(15)1002), the Agricultural Science and Technology Innovation Fund in Jiangsu Province, China (CX(12)1003-09), the National Key Research Program of China (2016YFD0300503), the Science and Technology Plan of Jiangsu Province, China (BE2015340), the Research Innovation Program for College Graduates of Jiangsu Province, China (KYLX15_1369), and a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, China.dry matter under PT were greater than those under CT and DS methods.  Besides, the entire growth duration and daily production showed significant positive relations with rice yield.  Compared with CT and DS, the effective accumulated temperature and cumulative solar radiation of rice under PT were higher in phenological phases.  In addition, the dry matter/effective accumulated temperature and solar energy utilization of rice under CT and DS were higher during vegetative phase and lower during reproductive and grain filling phases in contrast to PT.  The mean daily temperature and mean daily solar radiation in the entire growth duration showed significant positive correlation with rice yield, total dry matter, and harvest index.  This study demonstrated that when the mean daily temperature is <25.1°C in vegetative phase and >20.1°C in grain filling phase, rice yield could be increased by selecting mechanized planting methods.  Most varieties under PT method exhibited high yield and climate resources use efficiency compared with CT and DS.  In conclusion, the PT method could be a better cultivation measure for high rice yield, accompanied with high temperature and solar radiation use efficiency in a rice-wheat rotation system in the lower reaches of the Yangtze River, China.

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Susceptible time window and endurable duration of cotton fiber development to high temperature stress XU Bo, ZHOU Zhi-guo, GUO Lin-tao, XU Wen-zheng, ZHAO Wen-qin, CHEN Bing-lin, MENG Ya-li, WANG You-hua 2017, 16(09): 1936-1945.  DOI: 10.1016/S2095-3119(16)61566-6 Abstract ( )   PDF in ScienceDirect       The development of the cotton fiber is very sensitive to temperature variation, and high temperature stress often causes reduced fiber yield and fiber quality.  Short-term high temperature stress often occurs during cotton production, but little is known about the specific timing and duration of stress that affects fiber development.  To make this clear, pot experiments were carried in 2014 and 2015 in a climate chamber using cotton cultivars HY370WR (less sensitive variety) and Sumian 15 (heat sensitive variety), which present different temperature sensitivities.  Changes of the most important fiber quality indices (i.e., fiber length, fiber strength and marcironaire) and three very important fiber development components (i.e., cellulose, sucrose and callose) were analyzed to define the time window and critical duration to the high temperature stress at 34°C (max38°C/min30°C).  When developing bolls were subjected to 5 days of high temperature stress at different days post-anthesis (DPA), the changes (Δ%) of fiber length, strength and micronire, as a function of imposed time followed square polynomial eq. as y=a+bx+cx2, and the time around 15 DPA was the most sensitive period for fiber quality development in response to heat stress.  When 15 DPA bolls were heat-stressed for different durations (2, 3, 4, 5, 6, 7 days), the changes (Δ%) of fiber length, strength and micronire, as a function of stress duration followed logistic equations .  Referred to that 5, 10 and 15% are usually used as criteria to decide whether techniques are effective or changes are significant in crop culture practice and reguard to the fiber quality indices change range, we suggested that 5% changes of the major fiber quality indices (fiber length, fiber strength and micronaire) and 10% changes of fiber development components (cellulose, sucrose and callose) could be taken as criteria to judge whether fiber development and fiber quality have been significantly affected by high temperature stress.  The key time window for cotton fiber development in response to the high temperature stress was 13–19 DPA, and the critical duration was about 5 days.

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Characterization of salt tolerance and Fusarium wilt resistance of a sweetpotato mutant ZHANG Huan, ZHANG Qian, WANG Yan-nan, LI Yan, ZHAI Hong, LIU Qing-chang, HE Shao-zhen 2017, 16(09): 1946-1955.  DOI: 10.1016/S2095-3119(16)61519-8 Abstract ( )   PDF in ScienceDirect      The variant LM1 was previously obtained using embryogenic cell suspension cultures of sweetpotato variety Lizixiang by gamma-ray induced mutation, and then its characteristics were stably inherited through six clonal generations, thus this mutant was named LM1. In this study, systematic characterization of salt tolerance and Fusarium wilt resistance were performed between Lizixiang and mutant LM1. LM1 exhibited significantly higher salt tolerance compared to Lizixiang. The content of proline and activities of superoxide dismutase (SOD) and photosynthesis were significantly increased, while malonaldehyde (MDA) and H2O2 contents were significantly decreased compared to that of Lizixiang under salt stress. The inoculation test with Fusarium wilt showed that its Fusarium wilt resistance was also improved. The lignin, total phenolic, jasmonic acid (JA) contents and SOD activity were significantly higher, while H2O2 content was significantly lower in LM1 than that in Lizixiang. The expression level of salt stress-responsive and disease resistance-related genes was significantly higher in LM1 than that in Lizixiang under salt and Fusarium wilt stresses, respectively. This result provides a novel and valuable material for improving the salt tolerance and Fusarium wilt resistance of sweetpotato.

Horticulture

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Effect of ploidy level on expression of lycopene biosynthesis genes and accumulation of phytohormones during watermelon (Citrullus lanatus) fruit development and ripening DOU Jun-ling, YUAN Ping-li, ZHAO Sheng-jie, HE Nan, ZHU Hong-ju, GAO Lei, JI Wan-li, LU Xuqiang, LIU Wen-ge 2017, 16(09): 1956-1967.  DOI: 10.1016/S2095-3119(16)61618-0 Abstract ( )   PDF in ScienceDirect   The difference between lycopene and phytohormone levels among diploid, triploid and tetraploid plants of two watermelon cultivars during fruit growth and ripening was studied.  The expression pattern of five genes (phytoene synthase (PSY1), phytoene desaturase (PDS), ζ-carotene desaturase (ZDS), carotenoid isomerase (CRTISO), and lycopene β-cyclase (LCYB)) was analyzed in details.  In red-fleshed cultivar Mimei, lycopene content increased rapidly from 25 to 35 days after pollination (DAP), and then decreased at 40 DAP.  Triploid and tetraploid fruit had higher levels of lycopene than diploid.  Moreover, triploid tended to contain more lycopene than tetraploid during fruit growth and ripening stages.  However, little amount of lycopene (0–2 mg kg–1 fresh weight (FW)) in yellow-fleshed cultivar Huangmei was found during all fruit development stages.  In Mimei, transcript level of PSY1 was generally higher than the other four genes, and LCYB gene expression was the lowest among all five genes being tested.  PSY1, CRTISO and LCYB genes showed higher transcript levels in polyploid than in diploid fruit.  By contrast, in Huangmei, transcript level of LCYB was not the lowest, but only lower than that of PSY1.  PSY1, CRTISO and LCYB genes showed higher expression levels in diploid than in polyploid fruit.  In Mimei, the negative correlation between gibberellane (GA) content and lycopene accumulation was determined in all three different ploidy fruits, while a positive correlation was observed between abscisic acid (ABA) content and lycopene accumulation only in diploid watermelon.  These results indicated that different lycopene contents in different ploidy watermelons is regulated by the differential transcription expression of the lycopene metabolic genes and phytohormones.

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6-Benzylaminopurine treatment maintains the quality of Chinese chive (Allium tuberosum Rottler ex Spreng.) by enhancing antioxidant enzyme activity JIA Li-e, LIU Sheng, DUAN Xiao-ming, ZHANG Chao, WU Zhan-hui, LIU Ming-chi, GUO Shao-gui, ZUO Jin-hua, WANG Li-bin 2017, 16(09): 1968-1977.  DOI: 10.1016/S2095-3119(17)61663-0 Abstract ( )   PDF in ScienceDirect   Chinese chive usually develops an off-flavor after a short storage time.  To explore effective ways to maintain the postharvest quality of Chinese chive, the effect of exogenous application of 6-benzylaminopurine (6-BA) on postharvest quality and antioxidant activity of chive was evaluated, and the mechanism of the physiological responses of chive to 6-BA treatment was explored.  Chives were sprayed for 10 min with 100, 300, or 500 mg L–1 6-BA or with alkaline solution as the control, then stored at (2±1)°C with a relative humidity (RH) of 80–85%.  We found that 300 mg L–1 6-BA significantly delayed yellowing and chlorophyll degradation, maintained the total phenolic and flavonoid content, and improved the activities of antioxidant enzymes, including superoxide dismutase (SOD), catalase (CAT) and peroxidase (POD).  In conclusion, we identified exogenous application of 6-BA as an effective method for maintaining postharvest quality of Chinese chive.  In addition, our finding that the activities of antioxidant enzymes increase in response to exogenous 6-BA provides new insights into the mechanism of cytokinin-based postharvest fresh-keeping.

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Effect of continuous negative pressure water supply on the growth, development and physiological mechanism of Capsicum annuum L. LI Di, LONG Huai-yu, ZHANG Shu-xiang, WU Xue-ping, SHAO Hong-ying, WANG Peng 2017, 16(09): 1978-1899.  DOI: 10.1016/S2095-3119(16)61572-1 Abstract ( )   PDF in ScienceDirect   Effects of continuous negative pressure water supply on water consumption, growth and development, as well as physiological mechanism and quality of Capsicum annuum L. were investigated in this paper.  Meanwhile, the optimal negative pressure water supply conditions for growth of C. annuum L. were screened out to achieve the goals of water conservation, high yield and high quality, thus providing theoretical foundation for its field production.  The pot experiment within the greenhouse was utilized; the continuous negative pressure water supply was adopted; the four treatments, artificial watering (CK), –5 kPa (T1), –10 kPa (T2), and –15 kPa (T3) were set; and the daily water consumption, yield, as well as the biomass, nitrate reductase, root activity, vitamin C, capsaicin, and nutrient uptakes of nitrogen (N), phosphorus (P) and potassium (K) during various stages of its growth were determined.  Compared with CK, when the water supply pressure was controlled at –5 to –15 kPa in the experiment, the total water consumption of C. annuum L. reduced by 53.42 to 67.75%, the total water consumption intensity reduced by 54.29 to 67.14%, and the water use efficiency increased by 12.66 to 124.67%.  The N accumulation in a single strain of C. annuum L. from the color turning stage to the red ripe stage increased by 15.99 to 100.55%, respectively, compared with that of CK; the P accumulation increased by 20.47 to 154.00% relative to that of CK, and the K accumulation increased by 64.92 to 144.9% compared with that of CK.  Compared with CK, C. annuum L. yield was remarkably improved by 13.79% at T1, and contents of vitamin C, capsaicin as well as carotenoids at all growth stages were enhanced by 13.42–147.01%, 11.54–71.01%, and 41.1–568.06%, respectively.  Nitrate reductase activity, root activity and chlorophyll (a+b) were markedly increased by 335.78–500%, 79.6–140.68% and 114.95–676.19%, respectively, from immature stage to full ripe stage.  Adopting the continuous negative pressure water supply for C. annuum L. has a significant water-saving effect, and the water supply pressure being stable at –5 kPa contributes to its growth and development, improves yield, enhances root activity, promotes nutrient uptake, and improves its quality, thus achieving the effects of water conservation, high yield, high quality and high efficiency.

Plant Protection

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Study of viruses co-infecting white clover (Trifolium repens) in China LIANG Qiao-lan, WEI Lie-xin, XU Bing-liang, A. Calderón-Urrea, XIANG Dong 2017, 16(09): 1990-1998.  DOI: 10.1016/S2095-3119(16)61606-4 Abstract ( )   PDF in ScienceDirect   Globally, white clover (Trifolium repens L.) is commonly infected by plant viruses.  It is grown at gardens, roadsides, and public areas as ornamental plants in northern China.  Some leaves present disease symptoms that are similar to those of virus infection.  However, to our knowledge, no records are available from China regarding white clover (Trifolium repens L.) virus co-infection.  To determine the viral species that infect white clover in China, plant samples with virus disease symptoms were collected and virion morphology and ultrastructure morphology of co-infected plants were observed by electron microscopy; viruses were detected by enzyme linked immunosorbent assay (ELISA) and reverse transcription PCR (RT-PCR).  Virus co-infection was studied by double antibody sandwich enzyme linked immunosorbent assay (DAS-ELISA) and real-time fluorescence quantitative PCR (RT-qPCR) after rub-inoculation of virus-free white clover planted in a growth chamber (25°C) with a photoperiod of 16 h, using single or several purified virions.  Results showed that there were six types of symptoms, including those of shrinking mosaic, shrinking and macular mosaic, severe mottle mosaic, yellow macular mosaic, shrinkage chlorisis, and ring plaque and ring stria.  The incidence rates for each symptom were 20.93, 1.48, 16.85, 59.07, 1.30, and 0.37%, respectively, based on the field investigation.  Two types of viral pathogens were identified as Alfalfa mosaic virus (AMV) and White clover mosaic virus (WCMV).  In mesophyll cells, virus particles with bacilliform virions formed aggregates and linear virions were bundle shaped.  The detection rate of AMV was 100% in white clover samples by DAS-ELISA and RT-PCR detection, whereas that of WCMV was 83.33%.  The co-infection rate was 83.33%.  The relative contents of AMV and WCMV were significantly increased by 5.897- and 3.515-fold upon co-infection, when compared to that with single virus infection.  We observed larger starch particles and fewer or collapsed chloroplast grana in co-infected plants; in addition, vacuoles were twisted and smaller, compared to those of healthy plants.  To our knowledge, this is the first report of co-infection by AMV and WCMV in white clover, which has caused severe mosaicism and ultrastructure lesions in co-infected plants in China.

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Safe antifungal lipopeptides derived from Bacillus marinus B-9987 against grey mold caused by Botrytis cinerea GU Kang-bo, ZHANG Dao-jing, GUAN Cheng, XU Jia-hui, LI Shu-lan, SHEN Guo-min, LUO Yuanchan, LI Yuan-guang 2017, 16(09): 1999-2008.  DOI: 10.1016/S2095-3119(16)61616-7 Abstract ( )   PDF in ScienceDirect   Agricultural application studies, including field experiments and acute toxicity tests, were conducted for lipopeptides secreted by marine-derived Bacillus marinus B-9987.  Benefiting from commercially available scaled-up lipopeptide purification, the sample of impurities (isolated from target lipopeptides), raw extracted sample (purity: 9.08%), partially purified sample (purity: 20.86%), and highly purified sample (purity: 87.51%) were prepared from B. marinus B-9987 fermentation broth, and used in lab-scale antagonism tests, field experiments, swarming motility tests, and acute toxicity tests.  Operations and conditions in field experiments were consistent with the Pesticide-Guidelines for the Field Efficacy Trials (GB/T 17980.28-2000), and acute toxicity tests were executed according to Toxicological Test Methods of Pesticides for Registration (GB 15670-1995).  In agar diffusion tests in vitro and pot tests in vivo, all lipopeptide samples with different purities significantly inhibited Botrytis cinerea; meanwhile the sample of impurities isolated from target lipopeptides were not effective against B. cinerea.  Results of lab-scale tests showed that the target lipopeptides were effective substances against B. cinerea.  Thus, partially purified and raw extracted samples were used in field experiments instead of the highly purified sample for cost saving.  In the field experiments against rose grey mold, biological control efficacy of 500 mg L–1 lipopeptides reached 67.53%, slightly lower than 74.05% reached by the agrochemical pyrimethanil.  However, pyrimethanil severely suppressed B. marinus B-9987, whereas the lipopeptides promoted swarming motility and biocontrol efficacy of Bacillus biomass.  Lipopeptides at 87.51% purity were tested for systemic acute toxicity and confirmed as low-toxicity substances.  In conclusion, low-toxicity lipopeptides were potential alternatives to agrochemicals, and they also performed good promotion when combined with homologous biological control microorganism.  There were 2 breakthroughs in this research: (1) marine-derived bacterial lipopeptides inhibited grey mold caused by B. cinerea in field experiments; and (2) purified bacterial lipopeptides (sample purity: >87.51%) were determined to be low-toxicity substances by systemic acute toxicity tests, satisfying the strict requirement of pesticide registration in China (required purity: >85%).  This study provides support for using extracellular Bacillus-derived lipopeptides commercially similar to Bacillus-based biological control agents.

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Morphology and glucosinolate profiles of chimeric Brassica and the responses of Bemisia tabaci in host selection, oviposition and development LI Jun-xing, RAO Lin-li, XIE Hui, Monika Schreiner, CHEN Li-ping, LIU Yin-quan 2017, 16(09): 2009-2018.  DOI: 10.1016/S2095-3119(16)61617-9 Abstract ( )   PDF in ScienceDirect   Plant structures and chemicals, which are developed from the shoot apical meristem (SAM), form the main barriers to insect feeding.  A plant chimera containing cells of different genetic origins in the SAM will be morphologically and chemically different compared with the parents and thus may result in differential resistance to herbivores.  In this study, we explore if particular elements of plant resistance are localized in one of the layers of SAM; the replacement of one cell layer in a chimera may be linked to change of a single resistance trait to herbivores.  The morphology and glucosinolate profiles of two periclinal chimeras (labeled as TTC and TCC, respectively) and grafted parents tuber mustard (labeled as TTT) and red cabbage (labeled as CCC) were compared and the performance of whitefly (Bemisia tabaci) in host selection, oviposition preference and development were assessed under controlled conditions.  Both chimeras possessed leaf trichomes as parent tuber mustard TTT, however, TTC had significantly more trichomes than TCC and parent TTT.  Leaf wax content of both chimeras was intermediate between the two parents.  Five aliphatic and two indole glucosinolates were detected in both chimeras, whereas three aliphatic glucosinolates (3-methyl-sulfinylpropyl, 4-methyl-sulfinylbutyl and 2-hydroxy-3-butenyl) were not detected in tuber mustard, and one aliphatic glucosinolate (3-butenyl) was not detected in red cabbage.  Unexpectedly for a chimera, the quantities of two aliphatic glucosinolates (3-methyl-sulfinylpropyl and 4-methyl-sulfinylbutyl) in both TTC and TCC were 3- to 5-fold higher than parents.  In olfactory preference assays, B. tabaci showed preference to CCC, followed by TCC, TTC and TTT, and number of eggs laid showed the same pattern: CCC>TCC>TTC>TTT.  Interestingly, more whiteflies landed on TTT plants than the other three types in a free choice experiment and the developmental duration from egg to adult was the shortest on TTT and increased in the order TTT<TTC<TCC<CCC.  Our results indicate plant defenses traits of leaf waxes, trichomes and glucosinolates are not controlled by one cell layer of SAM, but are influenced by interactions amongst cell layers.  The overall findings suggest that periclinal chimera systems can be a valuable approach for the study of plant-insect interactions and may also be useful for future resistance breeding.

Animal Science · Veterinary Medicine

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MicroRNA-34c regulates porcine granulosa cell function by targeting forkhead box O3a XU Yuan, ZHANG Ai-ling, ZHANG Zhe, YUAN Xiao-long, CHEN Zan-mou, ZHANG Hao, LI Jia-qi 2017, 16(09): 2019-2028.  DOI: 10.1016/S2095-3119(16)61582-4 Abstract ( )   PDF in ScienceDirect   Granulosa cells (GCs) are somatic cells of ovary, the behaviors of GCs are important for ovarian function.  MicroRNAs (miRNAs) are a class of endogenous 18–24 nucleotide (nt) non-coding RNAs, some of which have been shown to be important regulators of GCs function.  miR-34c involved in the regulation of various biological processes and was identified to be a pro-apoptotic and anti-proliferative factor in many cell types.  However, the roles of miR-34c in GCs function remain unknown.  In this study, we used Annexin V-FITC and EdU assays to demonstrate that miR-34c exerted pro-apoptotic and anti-proliferative effects in porcine GCs.  Dual-luciferase reporter assays, quantitative real-time PCR (qRT-PCR) and Western blotting identified Forkhead box O3a (FoxO3a) as a direct target gene of miR-34c.  The overexpression of FoxO3a rescued the phenotypic change caused by miR-34c in porcine GCs.  In conclusion, miR-34c regulate the function of porcine GCs by targeting FoxO3a.

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Effect of yeast Saccharomyces cerevisiae supplementation on serum antioxidant capacity, mucosal sIgA secretions and gut microbial populations in weaned piglets ZHU Cui, WANG Li, WEI Shao-yong, CHEN Zhuang, MA Xian-yong, ZHENG Chun-tian, JIANG Zongyong 2017, 16(09): 2029-2037.  DOI: 10.1016/S2095-3119(16)61581-2 Abstract ( )   PDF in ScienceDirect   This study was conducted to determine the effect of different forms of yeasts Saccharomyces cerevisiae supplementation on serum antioxidant capacity, mucosal secretory immunoglobulin A (sIgA) secretions and gut microbial populations in weaned piglets.  A total of 96 piglets weaned at 14 d of age were randomly allotted to 4 dietary treatments: (1) basal diet without yeast (Control); (2) basal diet supplemented with 3.00 g kg–1 live yeast (LY); (3) basal diet supplemented with 2.66 g kg–1 heat-killed whole yeast (HKY); and (4) basal diet supplemented with 3.00 g kg–1 superfine yeast powders (SFY).  Each treatment had 4 replicates (pens), with 6 piglets per replicate.  The experiment lasted for 3 wk.  At d 7 and 21 of the experiment, the samples of serum, mucosa and mesenteric lymph node (MLN) from jejunum, and digesta from the ileum and cecum were collected for determinations.  Compared with the Control, dietary SFY supplementation increased serum superoxide dismutase (SOD) activity and lysozyme levels at d 7, and jejunum mucosal sIgA secretions at d 21 of the experiment (P<0.05).  Dietary LY supplementation increased serum SOD activity and jejunum mucosal sIgA secretions, but decreased serum malondialdehyde (MDA) concentration at d 7 and 21 (P<0.05).  Piglets fed diets supplemented with LY and SFY had lower pH values and decreased numbers of Escherichia coli in the ileum and cecum contents at d 21 compared with the Control (P<0.05).  Moreover, the ratio of Lactobacilli to E. coli in the ileum and cecum contents was increased by dietary LY and SFY supplementations (P<0.05).  Collectively, different forms of yeasts, especially LY and SFY, may modulate body antioxidant capacity and enhance the intestinal immunity by regulation of secretions of mucosal sIgA and reduction of pathogenic bacteria colonization, thus improving intestinal health of weaned piglets.

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Manganese enhances the expression of the manganese superoxide dismutase in cultured primary chick embryonic myocardial cells QIN Shi-zhen, LIAO Xiu-dong, LU Lin, ZHANG Li-yang, XI Lin, GUO Yan-li, LUO Xu-gang 2017, 16(09): 2038-2046.  DOI: 10.1016/S2095-3119(16)61527-7 Abstract ( )   PDF in ScienceDirect     In the present study, the effect of manganese (Mn) on antioxidant status and the expression of the manganese superoxide dismutase (MnSOD) gene in cultured primary myocardial cells collected from the chick embryos was investigated.  The hypothesis that Mn supplementation would enhance the expression of MnSOD in cultured primary myocardial cells of chick embryos was tested.  Eggs collected from Mn-depleted Arbor Acres laying breeder hens were incubated for 10 days and then myocardial cells were isolated and cultivated for 8 days.  The embryonic myocardial cells on day 6 were treated with Mn in the cell culture medium at different time points when the proportion of cells showing spontaneous contraction was over 95% after the 3-day primary culture.  A completely randomized design involving a 3 Mn levels (0, 0.5 and 1.0 mmol L–1)×3 incubation time points (12, 24 and 48 h) factorial arrangement of treatments (n=6) was used in the current experiment.  The results showed that MnSOD activity and mRNA expression level were induced by Mn and increased with incubation time, which supported the hypothesis that Mn would enhance the expression of the MnSOD gene, and thus might protect myocardial cells from oxidative stress during the chick embryonic development.

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Evaluation of an attenuated vaccine candidate based on the genotype C of bovine parainfluenza virus type 3 in albino guinea pigs MA Lei, ZHU Yuan-mao, YANG Ting, XUE Fei 2017, 16(09): 2047-2054.  DOI: 10.1016/S2095-3119(17)61661-7 Abstract ( )   PDF in ScienceDirect   Bovine parainfluenza virus type 3 (BPIV3) is considered as one of the most important respiratory tract pathogens of both young and adult cattle, and widespread among cattle in the world.  BPIV3 was first reported in China in 2008 and four strains of BPIV3 were isolated from Shandong Province, known as genotype C (BPIV3c).  Pathogen investigations had shown that BPIV3c infection was very common among cattle in China.  To date, BPIV3 can be classified into genotypes A, B and C based on genetic and phylogenetic analysis.  Serological survey also demonstrates that BPIV3 infection is widespread in China, however, there is still no available vaccine for BPIV3 prevention in China nowadays.  In the present study, the BPIV3c strain SD0835 was continuously passaged on Madin-Darby bovine kidney (MDBK) cells for hundreds of times, and the pathogenicity of passage 209 was reduced in guinea pigs.  The passage 209 of BPIV3c strain SD0835 was used as a live vaccine candidate to immunize the guinea pigs.  The vaccination results revealed that two vaccinations could induce excellent serum neutralizing antibody responses as well as proliferation of T lymphocytes.  The vaccinated guinea pigs were well protected against challenge with a low passage of BPIV3c strain SD0835.  Additionally, the percentages of CD4+ and CD8+ T cell subsets of animals in vaccinated group increased after immunization; T cell subsets on day 2 after challenge in both groups decreased, and the decline of CD4+ and CD8+ T cell subsets levels of four guinea pigs in vaccinated group was relatively moderate, comparing with that of the control group.  These data support further testing of the attenuated virus as an effective candidate vaccine.

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Development of a real-time RT-PCR method for the detection of newly emerged highly pathogenic H7N9 influenza viruses WANG Xiu-rong, GU Lin-lin, SHI Jian-zhong, XU Hai-feng, ZHANG Ying, ZENG Xian-ying, DENG Guo-hua, LI Cheng-jun, CHEN Hua-lan 2017, 16(09): 2055-2061.  DOI: 10.1016/S2095-3119(17)61655-1 Abstract ( )   PDF in ScienceDirect      In 2013, a human influenza outbreak caused by a novel H7N9 virus occurred in China.  Recently, the H7N9 virus acquired multiple basic amino acids at its hemagglutinin (HA) cleavage site, leading to the emergence of a highly pathogenic virus.  The development of an effective diagnostic method is imperative for the prevention and control of highly pathogenic H7N9 influenza.  Here, we designed and synthesized three pairs of primers based on the nucleotide sequence at the HA cleavage site of the newly emerged highly pathogenic H7N9 influenza virus.  One of the primer pairs and the corresponding probe displayed a high level of amplification efficiency on which a real-time RT-PCR method was established.  Amplification using this method resulted in a fluorescent signal for only the highly pathogenic H7N9 virus, and not for any of the H1–H15 subtype reference strains, thus demonstrating high specificity.  The method detected as low as 39.1 copies of HA-positive plasmid and exhibited similar sensitivity to the virus isolation method using embryonated chicken eggs.  Importantly, the real-time RT-PCR method exhibited 100% consistency with the virus isolation method in the diagnosis of field samples.  Collectively, our data demonstrate that this real-time RT-PCR assay is a rapid, sensitive and specific method, and the application will greatly aid the surveillance, prevention, and control of highly pathogenic H7N9 influenza viruses.

Agro-Ecosystem & Environment

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Comparative effects of nitrogen application on growth and nitrogen use in a winter wheat/summer maize rotation system YIN Min-hua, LI Yuan-nong, XU Yuan-bo 2017, 16(09): 2062-2072.  DOI: 10.1016/S2095-3119(16)61487-9 Abstract ( )   PDF in ScienceDirect   The application of fertilizer in agricultural production has become universally common for achieving high crop yields and economic benefits, but it has potential impacts on food safety, energy crisis and environmental pollution.  Optimal management of fertilization is thus necessary for maintaining sustainable agriculture.  Two-year (2013–2015) field experiment was conducted, in Yangling (108°24´E, 34°20´N, and 521 m a.s.l.), Shaanxi Province, China, to explore the effects of different nitrogen (N) applications on biomass accumulation, crop N uptake, nitrate N (NO3–-N) distribution, yield, and N use with a winter wheat/summer maize rotation system.  The N applications consisted of conventional urea (U) (at 80 (U80), 160 (U160), and 240 (U240) kg N ha–1; 40% applied as a basal fertilizer and 60% top-dressed at jointing stage) and controlled-release urea (CRU) (at 60 (C60), 120 (C120), 180 (C180), and 240 (C240) kg N ha–1; all applied as a basal fertilizer) with no N application as a control (CK).  The continuous release of N from CRU matched well with the N demands of crop throughout entire growing stages.  Soil NO3–-N content varied less and peaked shallower in CRU than that in urea treatments.  The differences, however, were smaller in winter wheat than that in summer maize seasons.  The average yield of summer maize was the highest in C120 in CRU treatments and in U160 in urea treatments, and apparent N use efficiency (NUE) and N agronomic efficiency (NAE) were higher in C120 than in U160 by averages of 22.67 and 41.91%, respectively.  The average yield of winter wheat was the highest in C180 in CRU treatments and in U240 in urea treatments with C180 increasing NUE and NAE by averages of 14.89 and 35.62% over U240, respectively.  The annual yields under the two N fertilizers were the highest in C120 and U160.  The results suggested that CRU as a basal fertilizer once could be a promising alternative of urea as split application in semiarid areas.

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Optimize nitrogen fertilization location in root-growing zone to increase grain yield and nitrogen use efficiency of transplanted rice in subtropical China WU Meng, LIU Ming, LIU Jia, LI Wei-tao, JIANG Chun-yu, LI Zhong-pei 2017, 16(09): 2073-2081.  DOI: 10.1016/S2095-3119(16)61544-7 Abstract ( )   PDF in ScienceDirect   The optimized nitrogen fertilization location differs in different rice-growing regions.  We optimized nitrogen deep-point application in root-growing zone (NARZ) for transplanted rice in subtropical China.  Field plot experiments were conducted over two years (2014–2015) in a double-rice cropping system to evaluate the effects of nitrogen (N) fertilizer location on grain yield and N use efficiency (NUE).  Four different nitrogen deep-point application methods (DN) were compared with traditional broadcast application (BN) using granular urea.  The results showed that grain yield,  recovery efficiency of N (REN), agronomic efficiency of N (AEN), and partial factor productivity of N (PFPN) significantly increased 10.3–63.4, 13.7–56.7, 24.7–201.9 and 10.2–63.4%, respectively, in DN treatment compared to BN, respectively.  We also find that DN treatments increased grain yield as well as grain N content, and thus grain quality, in comparison with conventional BN treatment.  Correlation analysis indicated that significant improvement in grain yield and NUE mainly resulted from increases in productive panicle number and grain N content.  In our proposed NARZ method, granular urea should be placed 0 to 5 cm around the rice seeding at a 12-cm depth druing rice transplanting.  In NARZ, balanced application of N, P and K further improved grain yield and NUE over treatments with a single N deep-point application.  High N uptake by the rice plant did not cause significant soil fertility depletion, demonstrating that this method could guarantee sustainable rice production.

Agro-ecosystem & Environment

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Nitrogen mobility, ammonia volatilization, and estimated leaching loss from long-term manure incorporation in red soil HUANG Jing, DUAN Ying-hua, XU Ming-gang, ZHAI Li-mei, ZHANG Xu-bo, WANG Bo-ren, ZHANG Yang-zhu, GAO Su-duan, SUN Nan 2017, 16(09): 2082-2092.  DOI: 10.1016/S2095-3119(16)61498-3 Abstract ( )   PDF in ScienceDirect   Nitrogen (N) loss from fertilization in agricultural fields has an unavoidable negative impact on the environment and a better understanding of the major pathways can assist in developing the best management practices. The aim of this study was to evaluate the fate of N fertilizers applied to acidic red soil (Ferralic Cambisol) after 19 years of mineral (synthetic) and manure fertilizer treatments under a cropping system with wheat-maize rotations. Five field treatments were examined: control (CK), chemical nitrogen and potash fertilizer (NK), chemical nitrogen and phosphorus fertilizer (NP), chemical nitrogen, phosphorus and potash fertilizer (NPK) and the NPK with manure (NPKM, 70% N from manure). Based on the soil total N storage change in 0–100 cm depth, ammonia (NH3) volatilization, nitrous oxide (N2O) emission, N plant uptake, and the potential N leaching loss were estimated using a mass balance approach. In contrast to the NPKM, all mineral fertilizer treatments (NK, NP and NPK) showed increased nitrate (NO3–-N) concentration with increasing soil depth, indicating higher leaching potential. However, total NH3 volatilization loss was much higher in the NPKM (19.7%) than other mineral fertilizer treatments (≤4.2%). The N2O emissions were generally low (0.2–0.9%, the highest from the NPKM). Total gaseous loss accounted for 1.7, 3.3, 5.1, and 21.9% for NK, NP, NPK, and NPKM treatments, respectively. Estimated N leaching loss from the NPKM was only about 5% of the losses from mineral fertilizer treatments. All data demonstrated that manure incorporation improved soil productivity, increased yield, and reduced potential leaching, but with significantly higher NH3 volatilization, which could be reduced by improving the application method. This study confirms that manure incorporation is an essential strategy in N fertilization management in upland red soil cropping system.

Food Science

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Effect of post-harvest UV-C irradiation and calcium chloride on enzymatic activity and decay of tomato (Lycopersicon esculentum L.) fruit during storage Saeideh Mansourbahmani, Behzad Ghareyazie, Sepideh Kalatejari, Reza Salehi Mohammadi, Vahid Zarinnia 2017, 16(09): 2093-2100.  DOI: 10.1016/S2095-3119(16)61569-1 Abstract ( )   PDF in ScienceDirect       Tomato is one of the extensively consumed vegetable crops worldwide.  The regular consumption of tomato decreases the incidence of chronic degenerative diseases such as certain types of cancer and cardiovascular diseases.  The objective of this study was to find an appropriate method that not only reduces tomatoes decay, but also maintains its post-harvest quality.  A factorial experiment based on randomized complete block design with three replications was conducted to evaluate effects of ultraviolet (UV)-C and CaCl2 applications on tomato during storage.  The traits studied included ethylene, polygalacturonase (PG) activity, pectin methyl esterase (PME) activity, firmness, total phenol content, and fungal-induced decay were measured weekly during 35 days of storage.  Both UV-C and CaCl2 treatments had positive effects on tomato quality as compared to control treatment.  The 3 and 4.5 kJ m–2 levels of UV-C and 2% CaCl2 had positive effects on quality characteristics, respectivly.  Fruits treated by UV-C and CaCl2 had higher phenol and firmness, and less PME activity, PG activity, ethylene production, and decay than the control fruits.  In conclusion, increasing in storage duration significantly affected the fruits quality by increasing in ethylene, PME activity, PG activity, decay and decreasing the phenol content and firmness.  But UV-C and CaCl2 led to significant decrease in this adverse impact relative to control treatment.

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Pesticide residues in bayberry (Myrica rubra) and probabilistic risk assessment for consumers in Zhejiang, China YANG Gui-ling, WANG Wen, LIANG Sen-miao, YU Yi-jun, ZHAO Hui-yu, WANG Qiang, QIAN Yongzhong 2017, 16(09): 2101-2109.  DOI: 10.1016/S2095-3119(16)61600-3 Abstract ( )   PDF in ScienceDirect       As pesticide residues in bayberry has raised serious concern from the public in China, a monitoring survey was carried out during 2013–2014 and 157 samples were analyzed in total.  Twenty-three pesticides were detected among the 44 pesticides analyzed and at least one pesticide was detected in 63% of 99 samples.  Whereas 45.6% of samples were found with two or more pesticide residues, and 23.6% of samples with four or more pesticide residues.  Probabilistic risk assessments indicated that estimated daily intake (EDI) of cyhalothrin at the P97.5th percentile level is 1.11 times larger than the acceptable daily intake (ADI) for children, the estimated short-term intake (ESTI) at the P97.5th percentile level is 1.9 and 1.78 times larger than the acute reference dose (ARfD) for adult and children, respectively, indicating the potential risk concern from pyrethroids.  EDI of the pesticides with anti-androgenic effects ranged from 0.15–2.46 of ADI, the probability of exposure exceeding the ADI was 7.1 and 31.1% for adults and children, respectively, the probability of exposure of pyrethroids exceeding the ADI was 3.8% for children.  Exposures for other pesticides and groups were below 1.0.  Because the co-occurrence of frequency of cyhalothrin and cypermethrin was 9.55%, the combinations of pesticides detected with anti-androgenic effects has not been found in the present monitoring, the results indicate that the pesticide residues in bayberry will not constitute serious public health risk, however, they are significant for the management of pesticide use on bayberry and dietary health risk in China.

Short Communication

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Screening of genes of secreting acetic acid from Aspergillus niger H1 to improve phosphate solubilization LIU Xue, ZHU Chang-xiong, Frank Delvigne, GONG Ming-bo 2017, 16(09): 2110-2114.  DOI: 10.1016/S2095-3119(16)61477-6 Abstract ( )   PDF in ScienceDirect       A primary cDNA library of Aspergillus niger H1 was constructed using the switching mechanism at the 5´ end of the RNA transcript (SMART) technique. A total of 169 clones exhibited halos when grown on tricalcium phosphate medium, and the H-46 clone displayed a clear halo. The full-length cDNA of the clone H-46 clone was 1 407 bp in length with a complete open reading frame (ORF) of 816 bp, and it encoded a protein that contained 272 amino acids. Multiple alignment analysis revealed a high degree of homology between the ORFs of the H-46 clone and the Bax inhibitor family (BI-1-like) proteins of other fungi. Acetic acid was secreted by Escherichia coli DH5α that express the BI-1-like gene. The level attained was 492.52 mg L−1, which was associated with the release of 0.212 mg mL−1 of soluble phosphate at 28 h. These results showed that the heterologous expression of BI-1-like genes in Eschericha coli DH5α increased the secretion of acetic acid by altering the membrane permeability and enhancing the solubility of phosphate (P).