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1. Comparative analysis of the genome of the field isolate V86010 of the rice blast fungus Magnaporthe oryzae from Philippines
ZHU Kun-peng, BAO Jian-dong, ZHANG Lian-hu, YANG Xue, LI Yuan, Zhu Ming-hui, LIN Qing-yun, ZHAO Ao, ZHAO Zhen, ZHOU Bo, LU Guo-dong
Journal of Integrative Agriculture    2017, 16 (10): 2222-2230.   DOI: 10.1016/S2095-3119(16)61607-6
摘要693)      PDF    收藏
Genome dynamics of pathogenic organisms are driven by plant host and pathogenic organism co-evolution, in which pathogen genomes are used to overcome stresses imposed by hosts with various genetic backgrounds through generation of a range of field isolates.  This model also applies to the rice host and its fungal pathogen Magnaporthe oryzae.  To better understand genetic variation of M. oryzae in nature, the field isolate V86010 from the Philippines was sequenced and analyzed.  Genome annotation found that the assembled V86010 genome was composed of 1 931 scaffolds with a combined length of 38.9 Mb.  The average GC ratio is 51.3% and repetitive elements constitute 5.1% of the genome.  A total of 11 857 genes including 616 effector protein genes were predicted using a combined analysis pipeline.  All predicted genes and effector protein genes of isolate V86010 distribute on the eight chromosomes when aligned with the assembled genome of isolate 70-15.  Effector protein genes are located disproportionately at several chromosomal ends.  The Pot2 elements are abundant in V86010.  Seven V86010-specific effector proteins were found to suppress programmed cell death induced by BAX in tobacco leaves using an Agrobacterium-mediated transient assay.  Our results may provide useful information for further study of the molecular and genomic dynamics in the evolution of M. oryzae and rice host interactions, and for characterizing novel effectors and AVR genes in the rice blast pathogen.  
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2. 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
Journal of Integrative Agriculture    2017, 16 (09): 1923-1935.   DOI: 10.1016/S2095-3119(16)61596-4
摘要1070)      PDF    收藏
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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3. 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
Journal of Integrative Agriculture    2016, 15 (10): 2299-2308.   DOI: 10.1016/S2095-3119(16)61413-2
摘要1550)      PDF    收藏
    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.
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4. In vitro Selection of DNA Aptamers and Fluorescence-Based Recognition for Rapid Detection Listeria monocytogenes
LIU Guo-qing, LIAN Ying-qi, GAO Chao, YU Xiao-feng, ZHU Ming, ZONG Kai, CHEN Xuejiao
Journal of Integrative Agriculture    2014, 13 (5): 1121-1129.   DOI: 10.1016/S2095-3119(14)60766-8
摘要2078)      PDF    收藏
Aptamers are specific nucleic acid sequences that can bind to a wide range of nucleic acid and non-nucleic acid targets with high affinity and specificity. Nucleic acid aptamers are selected in vitro from single stranded DNA or RNA ligands containing random sequences of up to a few hundred nucleotides. Systematic evolution of ligands by exponential enrichment (SELEX) was used to select and PCR amplify DNA sequences (aptamers) capable of binding to and detecting Listeria monocytogenes, one of the major food-borne pathogens. A simplified affinity separation approach was employed, in which L. monocytogenes in exponential (log) phase of growth was used as the separation target. A fluorescently-labeled aptamer assay scheme was devised for detecting L. monocytogenes. This report described a novel approach to the detection of L. monocytogenes using DNA aptamers. Aptamers were developed by nine rounds of SELEX. A high affinity aptamer was successfully selected from the initial random DNA pool, and its secondary structure was also investigated. One of aptamers named e01 with the highest affinity was further tested in aptamer-peroxidase and aptamer-fluorescence staining protocols. This study has proved the principle that the whole-cell SELEX could be a promising technique to design aptamer-based molecular probes for dectection of pathogenic microorganisms without tedious isolation and purification of complex markers or targets.
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