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Acta Biochimica et Biophysica Sinica 2005年第10期37卷 713-718页

作者： Ming LI Min SHAO Xu-Zhong LU Jin-Sheng WANG key laboratory of monitoring and management of plant diseases and Insects,Ministry of Agriculture,Nanjing Agricultural University,Nanjing 210095,China key laboratory of monitoring and management of plant diseases and Insects Ministry of Agriculture Nanjing Agricultural University Nanjing 210095 China

Many harpins have been found in plant pathogen bacteria that can elicit disease and insectresistance in plants,and promote plant *** this work,we overexpressed and purified Xanthomonasoryzae *** harpin,harpinXoo,in Escherichia coli BL21/*** fused to the C-terminus of glutathione S-transferase （GST） and purified using the Bulk GST purification module and thrombincleavage capture *** harpinXooprotein was sensitive to protease K and stable to heat treatment,andcould not induce a hypersensitive response after treatment with various plant metabolic inhibitors;thesecharacteristics were similar to harpinEaof Erwinia *** purified harpinXooshowed a similar abilityto induce tobacco mosaic virus resistance in tobacco as *** antibody worked well in detecting thepurified *** the total protein of *** BL21/pGEX-hpal and an hpal transgenic rice.

关键词： harpinXoo glutathione S-transferase (GST) biological activity polyclonal antibody

Synthesis, Biological Activity and 3D-QSAR Study of Novel Pyrrolidine-2,4-dione Derivatives Containing N-Substituted Phenylhydrazine Moiety

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Chemical Research in Chinese Universities 2015年第2期31卷 228-234页

作者： ZHANG Lizhi REN Zhengjiao LU Aimin ZHAO Zheng XU Wenqin BAO Qianqian DING Weijie YANG Chunlong Jiangsu key laboratory of Pesticide Science key laboratory of monitoring and management of Crop diseases and Pest lnsects Ministry of Agriculture College of Science Nanjing Agricultural University Nanjing 210095 P R. China

Twenty-seven novel pyrrolidine-2,4-dione derivatives containing N-substituted phenylhydrazine moiety were synthesized. Their structures were confirmed by IH NMR, 13C NMR and MS. The half effective concentration （ECs0） values of the title compounds against the phytopathogenic fungi Rhizoctonia cerealis were evaluated. Com- pounds 61 and 6q displayed good bioactivity with EC50 values of 1.626 and 2.043 μg/mL, respectively. The 3D quantitative structure activity relationship（3D-QSAR） model of CoMFA was established with reliable cross-validated correlation coefficient q2 value of 0.585 and Noncross-validated correlation coefficient r2 value of 0.971. This model provided a tool for guiding further design and synthesis of novel pyrrolidine-2,4-dione derivatives with high fungicidal activity.

关键词： Pyrrolidine-2,4-dione N-Substituted phenylhydrazine Antifungal activity 3D quantitative structure activity relationship(3 D-QSAR)

Effects of Temperature on Functional Response of Anagrus nilaparvatae Pang et Wang (Hymenoptera:Mymaridae) on the Eggs of Whitebacked planthopper,Sogatella furcifera Horvth and Brown planthopper,Nilaparvata lugens Stl

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Journal of Integrative Agriculture 2012年第8期11卷 1313-1320页

作者： MA Ming-yong PENG Zhao-pu HE Yuan Institute of plant Protection Hunan Academy of Agricultural Sciences Changsha 412002 P.R.China key laboratory of monitoring and management of plant diseases and lnsects Ministry of Agriculture Nanjing Agricultural University Nanjing 210095 P.R. China

Understanding the temperature affecting parasitic efficiency is critical to succeed in utilizing parasitoid as natural enemy in pest management. laboratory studies were carried out to determine the effects of temperature on parasitoid preference of female Anagrus nilaparvatae Pang et Wang （Hymenoptera：Mymaridae） to the eggs of whitebacked planthopper （WBPH）, Sogatella furcifera Horváth and brown planthopper （BPH）, Nilaparvata lugens Stl to build a composite model describing changes in parasitic response along a temperature gradient （18, 22, 26, 30, 34°C）. The results showed that attack responses of A. nilaparvatae on WBPH and BPH were the best described by a Type II functional response. The two parameters, attack rates （a） and handling times （Th）, of A. nilaparvatae to both eggs were influenced by the temperature. The maximum attack rates to WBPH （1.235） and BPH （1.049） were at 26 and 34°C, respectively, and the shortest handling times to WBPH （0.063） and BPH （0.057） were at 30 and 26°C, respectively. However, the optimal temperature for parasitic efficiency of A. nilaparvatae to WBPH and BPH eggs was both at 26°C, which showed that the present microclimate temperature of the habitat in the paddyfield was beneficial to A. nilaparvatae and indicated that parasitic efficiency of A. nilaparvatae would be impaired by global warming.

关键词： functional response temperature Anagrus nilaparvatae whitebacked planthopper brown planthopper

Gene cloning and expression of aminopeptidase N and cadherin from midgut of the rice stem borer, Chilo suppressalis

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Insect Science 2010年第5期17卷 393-399页

作者： Hong-Kun Yu Hao Ghen Yong-Jun Zhang Kong-Ming Wu Ge-Mei Liang Ze-Wen Liu Yu-Yuan Guo State key laboratory for Biology of plant diseases and Insect Pests Institute of Plant Protection Chinese Academy of Agricultural Sciences Beijing key laboratory of monitoring and management of Crop diseases and Pest Insects Ministry of Agriculture College of Plant Protection Nanjing Agricultural University Nanjing China

The rice stem borer, Chilo suppressalis Walker is one of the most important insect pests on rice in Asia, north Africa and southern Europe. Transgenic Bt rice has been developed in the laboratory with good resistance to this pest and other Lepidopteran insects, which will provide a possible alternative tool for this pest control. The full-length cDNAs encoding an aminopeptidase N （CsAPN） and a cadherin （CsCad） were cloned from C. suppressalis. CsAPN showed common features of, and high identities to, other insect APNs in its deduced amino acid sequence. Although a full-length cDNA encoding cadherin-like protein has been reported in GenBank, the newly isolated cadherin here （CsCad） showed some differences in its amino acid sequence, especially at the 7th cadherin repeat region （CR7）, which indicated the newly isolated CsCad might be another allele. CsAPN and CsCad were successfully expressed in insect Tn cells, and the blot analysis showed these two proteins could bind Bt toxin CrylAb. The results will provide valuable information for the studies of toxin mode of action and the possible toxin resistance mechanisms in this pest.

关键词： aminopeptidase N cadherin Chilo suppressalis heterologous expression,Tn cells

Tebufenozide resistance is associated with sex-linked inheritance in Plutella xylostella

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Insect Science 2015年第2期22卷 235-242页

作者： Guang-Chun Cao Zhao-Jun Han Entomology Department/key laboratory of monitoring and management of plant diseases and Insects Ministry of Agriculture NanjingAgricultural University Nanjing State key laboratory for Biology of plant Disease and Insect Pests Institute of Plant ProtectionChinese Academy of Agricultural Sciences Beijing China

The diamondback moth （DBM）, Plutella xylostella （L.）, is a major pest of cruciferous crops. Tebufenozide, a novel nonsteroidal ecdysone agonist, exhibits good efficacy and has played an increasingly important role in the control of Lepidopteran pests in China. For its resistance management, the genetic basis of tebufenozide resistance was studied using a laboratory selected resistant strain of DBM （resistant ratio, RR = 268）. A series of crosses with laboratory susceptible and resistant strains revealed that tebufenozide resistance in this pest was partially biased toward female heredity, with a large difference in RR for F1 （RR = 29） and rF1 progeny （RR = 147）. The dominance calculated for these 2 cross progeny was -0.788 and 0.09, respectively. Further analysis showed that the susceptible male and female larvae were similar in theft sensitivity to tebufenozide, but the resistant female larvae showed significantly higher resistance than the resistant male larvae. The heredity of tebufenozide resistance in DBM might be linked with the W sex chromosome, which suggested that DBM has the ability to develop high levels of resistance to tebufenozide. This is the first report of sex-linked inheritance oftebufenozide resistance in P xylostella （L.）.

关键词： insecticide resistance Plutella xylostella sex linkage tebufenozide

Release of active oxygen from phytopathogenic bacteria and their regulation

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Chinese Science Bulletin 1999年第1期44卷 71-75页

作者： LI Hongyu WANG Jinshengkey laboratory of monitoring and management of plant diseases and Pests, Ministry of Agriculture Department of plant Protection, Nanjing Agriculture University, Nanjing 210095, China Corresponding author key laboratory of monitoring and management of plant diseases and Pests Ministry of Agriculture Nanjing China Department of plant Protection Nanjing Agriculture University Nanjing China

Some phytopathogenic bacteria including Xanthomonas oryzae pv. oryzae （a rice leaf bacterial blight pathogen） and its virulent gene mutant XooM3105,Xanthomonas campestris pv. campestris, Erwinia carotovora subsp, carotovora and so on, could release active oxygen such as O2 and H2O2 by ***2+, NADPH and O2 concentration regulated the response. The response is sensitive to heat and proteinase. These results show that the component releasing active oxygen in the bacterial plant pathogen may involve special protein and enzyme and be secreted to culture liquid. Various components of rice tissue can significantly induce the release of active oxygen in Xanthomonas oryzae pv. oryzae.

关键词： phytopathogenic bacteria release of active oxygen rice bacterial leaf blight pathogen interaction between host and pathogen signal transduction.

Amplification and function analysis of N6-adenine-specific DNA methyltransferase gene in Nilaparvata lugens

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Journal of Integrative Agriculture 2016年第3期15卷 591-599页

作者： ZHANG Jiao XING Yan-ru HOU Bo-feng YUAN Zhu-ting LI Yao JIE Wen-cai SUN Yang LI Fei Department of Entomology College of Plant Protection Nanjing Agricultural UniversityKey Laboratory of Monitoring and Management of Plant Diseases and Insects Ministry of Agriculture key Lab of Agricultural Entomology Ministry of AgricultureInstitute of Insect Sciences Zhejiang University

Methylation of the N6 position of adenine, termed N6-methyladenine, protects DNA from restriction endonucleases via the host-specific restriction-modification system. N6-methyladenine was discovered and has been well studied in bacteria. N6-adenine-specific DNA methyltransferase（N6AMT） is the main enzyme catalyzing the methylation of the adenine base and knowledge of this enzyme was mainly derived from work in prokaryotic models. However, large-scale gene discovery at the genome level in many model organisms indicated that the N6AMT gene also exists in eukaryotes, such as humans, mice, fruit flies and plants. Here, we cloned a N6AMT gene from Nilaparvata lugens（Nlu-N6AMT） and amplified its fulllength transcript. Then, we carried out a systematic investigation of N6AMT in 33 publically available insect genomes, indicating that all studied insects had N6AMT. Genomic structure analysis showed that insect N6AMT has short introns compared with the mammalian homologs. Domain and phylogenetic analysis indicated that insect N6AMT had a conserved N6-adenine Mlase domain that is specific to catalyze the adenine methylation. Nlu-N6AMT was highly expressed in the adult female. We knocked down Nlu-N6AMT by feeding ds RNA from the second instar nymph to adult female, inducing retard development of adult female. In all, we provide the first genome-wide analysis of N6AMT in insects and presented the experimental evidence that N6AMT might have important functions in reproductive development and ovary maturation.

关键词： N6-adenine-specific DNA methyltransferase(N6AMT) Nilaparvata lugens expression RNA interference gene structure

Effect of Azoxystrobin on Oxygen Consumption and cyt b Gene Expression of Colletotrichum capsici from Chilli Fruits

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Agricultural Sciences in China 2009年第5期8卷 628-631页

作者： CHEN Yu JIN Li-hua ZHOU Ming-guo College of plant Protection Nanjing Agricultural University~Key Laboratory of Monitoring and Management of Crop Diseases and PestInsects Ministry of Agriculture Nanjing 210095 P.R. China

Azoxystrobin acts as an inhibitor of electron transport by binding to the Qo center of cytochrome b （cyt b）. Resistance to azoxystrobin was usually caused by the point mutation of cyt b gene or by the induction of alternative respiration. Oxygen consumption test for mycelia of Colletotrichum capsici showed that azoxystrobin inhibited mycelial respiration within 12 h; however, as time went on, the respiration of the mycelia recovered when the mycelia were treated with azoxystrobin and salicylhydroxamic acid （SHAM, a known inhibitor of alternative respiration）, and the oxygen consumption of the mycelia could not be inhibited. Meanwhile, cytochrome b （cyt b） gene expression increased with the recovery of mycelial respiration. The increased cyt b gene expression might play a role in the development of resistance to azoxystrobin in C. capsici.

关键词： QoI fungicides gene expression chili fungal respiration azoxystrobin resistance

Potential Distribution of Alien Invasive Species and Risk Assessment:a Case Study of Erwinia amylovora in China

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Agricultural Sciences in China 2007年第6期6卷 688-695页

作者： CHEN Chen CHEN Juan HU Bai-shi IIANG Ying-hua LIU Feng-quan key laboratory of monitoring and management of plant diseases and Insects Ministry of Agriculture Nanjing Agricultural UniversityNanjing 210095 P.R.China

Alien invasive species represent a severe risk to biodiversity and economy, as in the case of fire blight （Erwinia amylovora）, a bacterial disease that originated in North America, which may be released into new locations by means of fruit trade. On the basis of the knowledge of Erwinia amylovora＇s biophysical characteristics and environmental data, the geographic information system （GIS） has been applied to determine areas where Erwinia amylovora can potentially invade China. Temperature and precipitation, during the blossoming period, are considered to be two critical factors affecting the Erwinia amylovora＇s suitable climatic zones. This spatial modeling approach was validated from a case study in Europe, where the occurrence of Erwinia amylovora has been proven. The model prediction agreed with the occurrence of the bacteria recorded in Europe, and the same procedure has been applied to produce a potential establishment area in China＇s two preferential apple cultivation regions, Bohai Bay region and Huangtu Altiplano region. It has been found that areas belonging to the high-risk category are more or less the main apple producing areas, accounting for their great economic importance in China. This methodology provides an initial baseline for assessment, prevention, and management of alien species that may become invasive under certain environmental conditions. In addition, this modeling approach provides a tool for policy makers to use, in making decisions on management practices where alien species are involved.

关键词： biological invasions Erwinia amylovora GIS potential distribution risk assessment

Transgenic Rape with hrf2 Gene Encoding Harpin_(Xooc) Resistant to Sclerotinia sclerotinorium

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Agricultural Sciences in China 2008年第4期7卷 455-461页

作者： MA Ling-li HUO Rong GAO Xue-wen HE Dan SHAO Min WANG Qi key laboratory of monitoring and management of plant diseases and Pests Ministry of Agriculture/Department of Plant PathologyNanjing Agricultural University Nanjing 210095 P.R.China

The objective of this study was to increase the resistance of rape using the method of transformation. The gene hrf2 derived from Xathomonas oryzae pv. oryzicola encoding harpinxooc protein was constructed into transgenic vector pCAMBIA1301. The cotyledonal petiole segments from rapeseed variety Yangyou 4 were infected by Agrobacterium tumefaciens strain LBA4404/pCAMBIA1301-hrf2. Hygromycin-resistant green shoots were obtained. Successful integration of the foreign gene into the genome of the rapeseed variety Yangyou 4 was confirmed by PCR, RT-PCR, and β-glucuronidase analyses. Disease bioassays of transgenic plants revealed an improved resistance of transgenic plants to Rape sclerotiniose. In brief, the hrf2 gene can be transferred into rape using the method of Agrobacterium-mediated transformation, which increased the resistance to Sclerotinia sclerotinorium in the transgenic plant.

关键词： hrf2 gene Agrobacterium tumefaciens-mediated transformation transgenic oilseed rape resistance