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Apoplastic Proteases: Powerful Weapons against Pathogen Infection in Plants

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Plant Communications 2020年第4期1卷 44-53页

作者： Yan wang yuanchao wang Yiming wang Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095China

Plants associate with diverse microbes that exert beneficial,neutral,or pathogenic effects inside the *** the initial stages of invasion,the plant apoplast constitutes a hospitable environment for invading microbes,providing both water and *** response to microbial infection,a number of secreted proteins from host cells accumulate in the apoplastic space,which is related to microbial association or colonization ***,the molecular mechanisms underlying plant modulation of the apoplast environment and how plant-secreted proteases are involved in pathogen resistance are still poorly ***,several studies have reported the roles of apoplastic proteases in plant resistance against bacteria,fungi,and *** the other hand,microbe-secreted proteins directly and/or indirectly inhibit host-derived apoplastic proteases to promote *** findings illustrate the importance of apoplastic proteases in plant–microbe ***,understanding the protease-mediated apoplastic battle between hosts and pathogens is of fundamental importance for understanding plant–pathogen ***,we provide an overviewof plant–microbe interactions in the apoplastic *** define the apoplast,summarize the physical and chemical properties of these structures,and discuss the roles of plant apoplastic proteases and pathogen protease inhibitors in host–microbe *** and future perspectives for research into protease-mediated apoplastic interactions are discussed,which may facilitate the engineering of resistant crops.

关键词： apoplast protease protease inhibitor plant immunity plant-microbe interaction

Multiple-site mutations of phage Bp7 endolysin improves its activities against target bacteria

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Virologica Sinica 2015年第5期30卷 386-395页

作者： Can Zhang yuanchao wang Huzhi Sun Huiying Ren College of Animal Husbandry and Veterinary Medicine Qingdao Agricultural University

The widespread use of antibiotics has caused serious drug resistance. Bacteria that were once easily treatable are now extremely difficult to treat. Endolysin can be used as an alternative to antibiotics for the treatment of drug-resistant bacteria. To analyze the antibacterial activity of the endolysin of phage Bp7(Bp7e), a 489-bp DNA fragment of endolysin Bp7e was PCR-amplified from a phage Bp7 genome and cloned, and then a p ET28a-Bp7e prokaryotic expression vector was constructed. Two amino acids were mutated(L99A, M102E) to construct p ET28a-Bp7Δe, with p ET28a-Bp7e as a template. Phylogenetic analysis suggested that BP7e belongs to a T4-like phage endolysin group. Bp7e and its mutant Bp7Δe were expressed in Escherichia coli BL21(DE3) as soluble proteins. They were purified by affinity chromatography, and then their antibacterial activities were analyzed. The results demonstrated that the recombinant proteins Bp7e and Bp7Δe showed obvious antibacterial activity against Micrococcus lysodeikticus but no activity against Staphylococcus aureus. In the presence of malic acid, Bp7e and Bp7Δe exhibited an effect on most E. coli strains which could be lysed by phage Bp7, but no effect on Salmonella paratyphi or Pseudomonas aeruginosa. Moreover, Bp7Δe with double-site mutations showed stronger antibacterial activity and a broader lysis range than Bp7e.

关键词： bacteriophage Bp7 endolysin(Bp7e) Bp7 endolysin mu

Genetic relationships among Chinese and American isolates of Phytophthora sojaeassessed by RAPD markers

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Chinese Science Bulletin 2006年第17期51卷 2095-2102页

作者： wang Ziying wang yuanchao ZHANG Zhengguang ZHENG Xiaobuo Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095 China Department of biology Anhui Institute of Education Hefei 230061China

The genetic diversity of three geo- graphic populations of Phytophthora sojae from China and the United States was determined using random amplified polymorphic DNA (RAPD). The purpose was to explore genetic relationships among Chinese and American isolates of the organism. 21 random primers were selected among 200 random primers screened. A total of 223 reproducible RAPD fragments were scored among 111 individuals, of which 199 (89.23%) were polymorphic. Analysis of genetic variation showed that there existed higher genetic variation in the United States population in comparison to the Chinese populations. Nei’s genetic identity and principal component analysis indicated that the populations of Fujian and United States are closer to each other than to Heilongjiang populations. Shannon-Wiener diversity index revealed that the United States populations have a higher genetic di- versity than that of Chinese populations. These data are in support of the hypothesis that P. sojae in the United States might not have been introduced from China.

关键词：植物生理学遗传多样性地理分布 RAPD

Large chromosomal segment deletions by CRISPR/LbCpf1-mediated multiplex gene editing in soybean

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Journal of Integrative Plant Biology 2021年第9期63卷 1620-1631页

作者： Kaixuan Duan Yuanyuan Cheng Jing Ji Chenchen wang Yongshu Wei yuanchao wang Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095China

The creation of new soybean varieties has been limited by genomic duplication and *** multiplex gene editing and large chromosomal segment deletion through clustered regularly interspaced palindromic repeats(CRISPR)/CRISPR-associated protein(Cas)systems are promising strategies for overcoming these ***/Cpf1 is a robust tool for multiplex gene ***,large chromosomal excision mediated by CRISPR/Cpf1 has been reported in only a few non-plant ***,we report on CRISPR/LbCpf1-induced large chromosomal segment deletions in soybean using multiplex gene *** CRISPR/LbCpf1 system was optimized for direct repeat and guide RNA lengths in crispr RNA(crRNA)*** editing efficiency was evaluated using LbCpf1 driven by the CaMV35S and soybean ubiquitin *** optimized system exhibited editing efficiencies of up to 91.7%.Our results showed eight gene targets could be edited simultaneously in one step when a single eight-gRNA-target crRNA array was employed,with an efficiency of up to 17.1%.We successfully employed CRISPR/LbCpf1 to produce small fragments(<1 Kb)and large chromosomal segment deletions(10 Kb-1 Mb)involving four different gene clusters in ***,these data demonstrate the power of the CRISPR/LbCpf1 platform for multiplex gene editing and chromosomal segment deletion in soybean,supporting the use of this technology in both basic research and agricultural applications.

关键词： chromosomal segment deletion CRISPR/LbCpf1 multiplex gene editing soybean

A Phytophthora sojae gene of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) induced in host infection and its anti-oxidative function in yeast

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Chinese Science Bulletin 2006年第11期51卷 1316-1323页

作者： ZENG Juan wang yuanchao SHEN Gu ZHENG Xiaobo Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 China

Glyceraldehyde-3-phosphate dehydro- genase (GAPDH) is a multifunctional protein well defined in eukaryotes, especially in mammalian and Saccharomyces cerevisiae. Using the method of suppression subtractive hybridization (SSH), we identified a Phytophthora sojae cDNA coding GAPDH, which was up-regulated during the early stage of soybean infection. The termed PsGapdh gene pos- sessed three copies in the P. sojae genome. Its amino acid sequence harbored overall conserved domain of GADPH, homologous closest to GapC1 of Achlya bisexualis (oomycete) and adjoined to GapC2s of Odontella sinensis and Phaeodactylum tricornutum (diatom), on the C-Ⅱ branch of subfamily GapC in phylogeny tree of GAPDH. The transcrip- tional level of PsGapdh was up-regulated throughout early infection. Heterogenous expression of PsGapdh in the yeast tdh1-deleted mutant could rescue growth arrest under continuous exposure to H2O2. These results indicated active roles of PsGapdh in patho- gen-host interaction and anti-oxidation.

关键词：甘油醛磷酸盐脱氢酶 GAPDH 病原体-宿主相互作用抗氧化

The Phytophthora effector Avh94 manipulates host jasmonic acid signaling to promote infection

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Journal of Integrative Plant Biology 2022年第11期64卷 2199-2210页

作者： Yao Zhao Bo Yang Huawei Xu Jinbin Wu Zhiyang Xu yuanchao wang Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095China College of Grassland Science Nanjing Agricultural UniversityNanjing 210095China

The oomycete pathogen Phytophthora sojae is a causal agent of soybean root *** colonization of soybeans,*** secretes various RXLR effectors to suppress host immune responses,supporting successful *** research has demonstrated that the RXLR effector Avh94 functions as a virulence effector,but the molecular mechanism underlying its role in virulence remains ***,we demonstrate that Avh94 overexpression in plants and pathogens promotes Phytophthora ***94 interacts with soybean JAZ1/2,which is a repressor of jasmonic acid(JA)***94 stabilizes JAZ1/2 to inhibit JA signaling and silencing of JAZ1/2 enhances soybean resistance against ***,*** lines overexpressing Avh94 inhibit JA ***,exogenous application of methyl jasmonate improves plant resistance to *** together,these findings suggest that *** employs an RXLR effector to hijack JA signaling and thereby promote infection.

关键词： effector JA signaling Phytophthora pathogen plant immunity

CRISPR/Cas9-mediated editing of GmTAP1 confers enhanced resistance to Phytophthora sojae in soybean

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Journal of Integrative Plant Biology 2023年第7期65卷 1609-1612页

作者： Tengfei Liu Jing Ji Yuanyuan Cheng Sicong Zhang Zeru wang Kaixuan Duan yuanchao wang Department of Plant Pathology Nanjing Agricultural UniversityNanjing 210095China

Soybean(Glycine max) is an economically important oilbearing crop that is widely planted worldwide(Babu et al.,2017). Many diseases seriously limit soybean yield. Soybean root rot disease, one of the most destructive ... 详细信息

关键词： soybean CRISPR/Cas9 planted

Responses of ephemeral plant germination and growth to water and heat conditions in the southern part of Gurbantunggut Desert

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Chinese Science Bulletin 2006年第A01期51卷 110-116页

作者： wang Xueqing JIANG Jin wang yuanchao LUO Weilin SONG Chunwu CHEN Junjie Xinjiang Institute of Ecology and Geography Chinese Academy of Sciences Urumqi 830011 China Xinjiang Administration of Engineering Construction in Erqis River Basin Urumqi 830000 China Graduate School of Chinese Academy of Sciences Beijing 100039 China

在 Gurbantunggut 沙漠的南部的部分的短暂植物系统地从 2002 ～ 2004 被监视，气象学的数据和土壤潮湿在一样的时期期间被分析。结果证明短暂植物萌芽和生长对水和热状况的变化敏感。超过 0 的每日的温度的时间 ?????????

关键词：短生植物水热环境 Gurbantunggut沙漠积极响应

Green fluorescent protein (GFP) as a vital marker for studying the interaction of Phytophthora sojae and soybean

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Chinese Science Bulletin 2009年第16期54卷 2822-2829页

作者： CHEN XiaoRen CHENG BaoPing wang XinLe DONG SuoMeng wang YongLin ZHENG XiaoBo wang yuanchao Department of Plant Pathology Nanjing Agricultural University Nanjing 210095 China

Transgenic Phytophthora sojae strains that produce green fluorescent protein (GFP) were obtained after stable DNA integration using the Hsp70 promoter and the Ham34 terminator of Bremia lactucae. The expression of GFP during different developmental stages of P. sojae was observed using fluorescent microscopy. Based on this reporter system, the histopathologic events caused by the pathogen in soybean leaves, hypocotyls and roots were monitored. Meanwhile, the difference in resistance between different soybean cultivars against P. sojae was analyzed microscopically in roots. The results indicate that GFP can be stably expressed in zoosporangia, zoospores, cysts, hyphae and oospores of P. sojae. Using the GFP marker, the infecting pathogens in leaves, hypocotyls and roots of host could be distinctly visualized. The germ tube length of cysts germinating on the roots of resistant cultivar Nannong 8848 was longer than that on the roots of susceptible cultivar Hefeng 35. These results show for the first time that this eukaryotic reporter can be used in P. sojae as a stable and vital marker, allowing the study of genetics of this hemibiotrophic pathogen.

关键词：绿色荧光蛋白标记大豆疫霉根腐病 GFP 大豆疫霉菌热休克蛋白70 显微镜观察大豆叶片 DNA整合