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Functional identification of phenazine biosynthesis genes in plant pathogenic bacteria Pseudomonas syringae pv. tomato and xanthomonas oryzae pv. oryzae

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Journal of Integrative Agriculture 2016年第4期15卷 812-821页

作者： LI Wen XU You-ping Jean-Pierre Munyampundu XU Xin QI Xian-fei GU Yuan CAI Xin-zhong Institute of Biotechnology College of Agriculture and Biotechnology Zhejiang University Center of Analysis and Measurement Zhejiang University State Key Laboratory of Rice Biology Zhejiang University

Phenazines are secondary metabolites with broad spectrum antibiotic activity and thus show high potential in biological control of pathogens. In this study, we identified phenazine biosynthesis （phz） genes in two genome-completed plant pathogenic bacteria Pseudomonas syringae pv. tomato （Pst） DC3000 and xanthomonas oryzae pv. oryzae （Xoo） PXO99A. Unlike the phz genes in typical phenazine-producing pseudomonads, phz homologs in Pst DC3000 and Xoo PXO99A consisted of phzC/D/E/F/G and phzC/E1/E2/F/G, respectively, and the both were not organized into an operon. Detection experiments demonstrated that phenazine-l-carboxylic acid （PCA） of Pst DC3000 accumulated to 13.4 IJg L-1, while that of Xoo PXO99A was almost undetectable. Moreover, Pst DC3000 was resistant to 1 mg mL-1 PCA, while Xoo PXO99A was sensitive to 50 IJg mL ~ PCA. Furthermore, mutation of phzF blocked the PCA production and significantly reduced the pathogenicity of Pst DC3000 in tomato, while the complementary strains restored these phenotypes. These results revealed that Pst DC3000 produces low level of and is resistant to phenazines and thus is unable to be biologically controlled by phenazines. Additionally, phz-mediated PCA production is required for full pathogenicity of Pst DC3000. To our knowledge, this is the first report of PCA production and its function in pathogenicity of a plant pathogenic P. syringae strain.

关键词： pathogenicity phenazine biosynthesis genes phenazine-l-carboxylic acid plant pathogenic bacteria,Pseudomonas syringae pv. tomato xanthomonas oryzae pv. oryzae

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Evaluation of the Pathotypes of xanthomonas oryzae pv. oryzae and Preliminary Analysis of the Resistant Reactions of Main Japonica Rice in the Yunnan Plateau, China

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Agricultural Sciences in China 2006年第4期5卷 299-306页

作者： YU Teng-qiong LIANG Bin YE Chang-rong XU Fu-rong TANG Cui-feng MAO Xiao-qiang DAI Lu-yuan Insititute of Biotechnology and Germplasm Resources Yunnan Academy of Agricultural Sciences Kunming 650205 P.R.China College of Agronomy and Biotechnology Yunnan Agricultural University. Kunming 650201 P.R.China

The pathogenicity of 36 isolates of xanthomonas oryzae pv. oryzae （Xoo）, which were collected from japonica rice varieties in the Yunnan Plateau, China, was evaluated. It was evaluated on 29 rice varieties including a set of seven varieties to identify pathogenicity, i.e., Haonuoyang, TN1, Kogyoku, Zhenzhu＇ai, IR26, Nanjing 33, and Kinmaze, which may be considered as a set of differential varieties for Xoo races from Yunnan japonica rice. The efficiency of the seven varieties was further confirmed. The results showed reversible and specific interactions between isolates and varieties. The isolates were classified into nine pathotypes from pathotyp Ⅰ to Ⅸ according to their pathogenic reactions on the seven rice varieties. The pathotype V was the epidemic, whereas pathogen Ⅶ was the most pathogenic. Most japonica varieties grown in the Yunnan Plateau were susceptible to Xoo. The rice lines IRBB21 （Xa-21）, Zhachanglong （Xa-22,, Xa- 24,）, and IR1545-339 （xa-5）, which were resistant to all the isolates tested, can be used as donors of resistant genes for bacterial blight in japonica rice breeding in the Yunnan Plateau.

关键词： japonica rice xanthomonas oryzae pv. oryzae pathotype resistance Yunnan

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Virulence of xanthomonas oryzae pv. oryzae on Rice Near-Isogenic Lines with Single Resistance Gene and Pyramiding Lines in China

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Agricultural Sciences in China 2004年第10期3卷 764-769页

作者： LIUHong-xia LIUFeng-quan HUBai-shi YANGWan-feng CHENZhi-yi XUZhi-gan DepartmentofPlantPathology NanjingAgriculturalUniversityNanjing210095P.R.China InstituteofPlantProtection JiangsuAcademyofAgriculturalSciencesNanjng210014P.R.China

Ninety one isolates of xanthomonas oryzae pv. oryzae were collected from different rice- growing regions in China and determined for their virulence on 24 rice near-isogenic lines containing single resistance gene and 2-4 genes: IRBB1 (Xa1), IRBB2 (Xa2), IRBB3 (Xa3), IRBB4 (Xa4), IRBB5 (xa5), IRBB7 (Xa7), IRBB8 (xa8), IRBB10 (Xa10), IRBB11 (Xa11), IRBB13 (xa13), IRBB14 (Xa14), IRBB21 (Xa21), IR24 (Xa18), IRBB50 (Xa4 + xa5), IRBB51 (Xa4 + xa13), IRBB52 (Xa4 + Xa21), IRBB53 (xa5 + xa13), IRBB54 (xa5 + Xa21), IRBB55 (xa13 + Xa21), IRBB56 (Xa4 + xa5 + xa13), IRBB57 (Xa4 + xa5 + Xa21), IRBB58 (Xa4 + xa13 + Xa21), IRBB59 (xa5 + xa13 + Xa21) and IRBB60 (Xa4 + xa5 + xa13 + Xa21). The results showed that most isolates were less virulent on lines with more than one genes pyramided than those with single resistance gene. The isolates tested were more virulent on IR24 and IRBB10, less virulent on IRBB5, IRBB7 and IRBB21. Based on interactions between isolates and rice near-isogenic lines, 7 cultivars with single gene (IRBB5, IRBB4, IRBB3, IRBB14, IRBB2, IRBB1 and IR24) were chosen as the differentials, and the tested isolates were classified into 7 virulence groups. The reaction patterns of the 7 groups in order were: RRRRRRR, RRRRRRS, RRRRRSS, RR/SRRSSS, RRRSSSS, RRSSSSS, RSSSSSS. The virulence frequencies were 7.69, 6.59, 14.29, 12.09, 14.29, 28.57 and 16.48% respectively. The elementary system for races identification has been established in China based on the results. It will be possible to compare with races in other countries, and the results will facilitate the development of rice resistance breeding to bacterial blight in China.

关键词： Rice lines xanthomonas oryzae pv. oryzae Virulence Resistance gene

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AvrXa3: A novel member of avrBs3 gene family from xanthomonas oryzae pv. oryzae has a dual function

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Progress in Natural Science:Materials International 2004年第9期 30-36页

作者： LI Ping*, LONG Juying, HUANG Yingchun, ZHANG Yan and WANG Jinsheng**(Department of Plant Pathology, Nanjing Agricultural University, Key Laboratory of Monitoring and Management of Plant Disease and Insects, Ministry of Agriculture, Nanjing 210095, China) Department of Plant Pathology Nanjing Agricultural University Key Laboratory of Monitoring and Management of Plant Disease and Insects Ministry of Agriculture Nanjing 210095 China

Two positive clones pUAV45 and pUAV47 were identified from the cDNA library of JxoⅢ, a race 3 strain of xanthomonas oryzae pv. oryzae Dye (Xoo) in Japanese system, using Tn5 based technology. pUAV45 clone contained a 25.4 kb Xoo genomic DNA insert. Southern blot analysis with avrXa 10 as the probe showed that DNA insert in pUAV45 shares homology with avrX 10. Furthermore within the Xoo insert, a smaller 5.7 kb Kpn I fragment (pUAV5k) was identified through hybridization with avrXa 10. The transformation of pUAV45 and pUAV5k into the strain Pxo99 (race 6 in Philippine system) led to the decrease of Pxo99 pathogenicity on rice cultivar Wase Aikoku 3 ( Xa 3) and the increase of the pathogen pathogenicity on Cas209 ( Xa 10). The result of sequence analysis showed that there is a 2598 bp open reading frame (ORF) within the 5.7 kb Kpn 1 fragment (pUAV5k). The ORF shared high identity (97%) with avrXa 10. The deduced sequence of the ORF contained 8.5 tandem repeat units of 34 amino acids, one leucine zipper (LZ), three nuclear localization signal (NLS) motifs, and an acidic activation transcriptional domain (AAD) at C terminus. We named this ORF avrXa 3 and it is classified as a new member of avrBs 3 ( avr/pth ) family with the dual function determined by alternations of avirulence and aggressiveness on rice cultivars carrying different ‘ R ’ genes.

关键词： rice xanthomonas oryzae pv. oryzae avrXa3 avrBs3 ( avr/pth ) family functional domains.

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Molecular Screening of Rice Cultivated in Benin for the Identification of xanthomonas oryzae pv. oryzae and Bacterial Leaf Blight Resistance Genes

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Advances in Bioscience and Biotechnology 2023年第12期14卷 514-533页

作者： Chimène Nadège Mahoussi Nanoukon Koffi David Montcho Hambada Antoine Abel Missihoun Kéllya Laurinzo Déguénon Bignon Meyrix Pamela Franzel Loumédjinon Bana Wêtè Déré Félicité Bio Emilienne Zinsou Réel Gael Fael Houngbélagnon Amed Sèmèvo Havivi Lamine Baba-Moussa Lambert Gustave Djédatin Laboratory of Molecular Biology and Bioinformatics Applied to Genomics National School of Applied Biosciences and Biotechnolo-gies National University of Science Technology Engineering and Mathematics Dassa-Zoumè Benin Laboratory of Molecular Biology and Bioinformatics Applied to Genomics National School of Applied Biosciences and Biotechnologies National University of Science Technology Engineering and Mathematics Dassa-Zoumè Benin Laboratory of Plant Horticultural and Forestry Sciences National University of Agriculture Kétou Benin Laboratory of Molécular Genetics and Genome Analysis Department of Genetics and Biotechnology Faculty of Science and Technology (FAST) University of Abomey-Calavi Cotonou Benin Laboratory of Biology and Molecular Typing in Microbiology Department of Biochemistry and Cell Biology University of Abomey-Calavi Abomey-Calavi Benin

One of the most devastating diseases of rice worldwide is bacterial blight (BLB) caused by xanthomonas oryzae pv. oryzae (Xoo). In Benin, Xoo was first described in 2013 on wild rice oryzae longistaminata. So far, no study has been done on Beninese Xoo strains. We do not know whether the pathogen has already passed into the rice varieties grown, or if they are exposed to other bacteria. Whereas the use of resistant varieties, carrying resistance genes, is the only highly effective and environmentally friendly way to control this disease, no information is available on these Xoo resistance genes in rice varieties grown in Benin apart from the one we recently. This study aims to identify Beninese Xoo strains, causing BLB and screen rice varieties grown in Benin for the main resistance genes. Diseased rice leaves showing typical symptoms of fire blight collected from different rice fields in the three phytogeographic areas of Benin were analyzed by PCR for Xoo-specific sequence identification. Furthermore, seventy-five collected rice accessions were screened to identify xa5, Xa7, xa13, and Xa21 resistance genes to Xoo. The results reveal that xanthomonas oryzae was identified in two fields in Banikouara and one in Malanville. On the other hand, Sphingomonas sp. has been identified in several other rice fields in Benin. Forty-seven of seventy-five rice accessions examined (62.66%) carried Xoo resistance genes with 3 (4%) and 40 (53.33%) of xa5 and Xa21 respectively. None of the accessions had either Xa7 or xa13 resistance genes. Three accessions possess both xa5 and Xa21 genes. Isogenic lines IRBB60 and IRBB21, supposed to be a positive control, presented a Xoo sensitivity allele. These results indicate that Xoo has moved from the wild rice variety to the cultivated variety in northern Benin and varietal improvement programs must be implemented with varieties having several resistance genes for the efficient response against a possible BLB pandemic in Benin.

关键词： Bacterial Blight xanthomonas oryzae pv. oryzae Molecular Characterization Resistance Genes

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Differential Detection and Analysis of Pathotypes and Differentiation against xanthomonas oryzae pv. oryzae in Southern China

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Agricultural Biotechnology 2018年第1期7卷 92-96页

作者： Shen CHEN Congying WANG Jing SU Aiqing FENG Xiaoyuan ZHU Liexian ZENG Guangdong Provincial Key Laboratory of High Technology for Plant Protection Plant Protection Research InstituteGuangdong Academy of Agricultural Sciences

Pathogenic isolates were collected from different rice-cropping regions in southern China to dissect the pathogenic disintegration and variation of Xanthornonas oryzae pv. oryzae （Xoo）. Two sets of rice-Xoo differential hosts, including Chinese system with such five euhivars as IR26, Java14, Nangeng15, Tetep and Jingang 30 and international system with a series of nearisogenic lines （NILs） including IRBB5, IRBB13, IRBB3, IRBB14, IRBB2 and IR24 carrying different known resistance genes,were used to detect pathogenic disintegration for the reaction between host and pathogen with leaf-clippingmethod at the rice booting stage. The results showed the type of the pathogen were divided intosix pathotypes, i.e. , Ⅰ, Ⅱ, Ⅲ, Ⅳ, Ⅴ and Ⅸ based on the Chinese differential system, and seven pathogenicraces including RI, R2, R3, R4, R5, R8 and R10 based on the international differential system. The pathogenicity frequency of Xoo pathotypes V and IV and pathogenic races R8 and R5 were 27.40% , 19.30% and 44.67% , 15.34%, which were considered to be the prevailing races in southern China. Pathogenic rates of pathotypes Ⅳ, Ⅴ, Ⅸ and races R8, R5 against 500 varieties derived from southernChina were 96.40% , 95.00% , 50.40% , 62.00% , and 42.60%, respectively. Among which pathotype Ⅸ was the most virulent pathotype. The pathotype Ⅴ became preponderant pathotype and the new pathotype Ⅸ grew up quickly.

关键词： xanthomonas oryzae pv. oryzae Differential variety Pathotype Race

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Recent Advances in Cloning and Characterization of Disease Resistance Genes in Rice

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Journal of Integrative Plant Biology 2007年第1期49卷 112-119页

作者： Liang-Ying Dai Xiong-Lun Liu Ying-Hui Xiao Guo-Liang Wang Rice Genomics Laboratory Hunan Agricultural University Changsha 410128 China Department of Plant Pathology The Ohio State University Columbus OH 43210 USA

Rice diseases caused by fungi, bacteria and viruses are one of the major constraints for sustainable rice （Oryza sativa L.） production worldwide. The use of resistant cultivars is considered the most economical and effective method to control rice diseases. In the last decade, a dozen resistance genes against the fungal pathogen Magnaporthe grisea and the bacterial pathogen xanthomonas oryzae pv. oryzae have been cloned. Approximately half of them encode nuclear binding site （NBS） and leucine rich repeat （LRR）-containing proteins, the most common type of cloned plant resistance genes. Interestingly, four of them encode novel proteins which have not been identified in other plant species, suggesting that unique mechanisms might be involved in rice defense responses. This review summarizes the recent advances in cloning and characterization of disease resistance genes in rice and presents future perspectives for in-depth molecular analysis of the function and evolution of rice resistance genes and their interaction with avirulence genes in pathogens.

关键词： rice blast bacterial blight Oryza sativa Magnaporthe grisea xanthomonas oryzae pv. oryzae

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Genetics and Improvement of Bacterial Blight Resistance of Hybrid Rice in China

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Rice science 2009年第2期16卷 83-92页

作者： ZHANG Qi Institute of Crop Sciences Chinese Academy of Agricultural Sciences Beijing 100081 China

Since 1980s, rice breeding for resistance to bacterial blight has been rapidly progressing in China. The gene Xa4 was mainly used in three-line indica hybrid and two-line hybrid rice. The disease has been ＇quiet＇ for 20 years in China, yet in recent years it has gradually emerged and been prevalent in fields planted with newly released rice varieties in the Changjiang River valley. Under the circumstances, scientists inevitably raised several questions： what causes the resurgence and what should we do next？ And/or is resistance breeding still one of the main objectives in rice improvement？ Which approach do we take on resistance breeding so that the resistance will be more durable, and the resistance gene will be used more efficiently？ A combined strategy involving traditional method, molecular marker-assisted selection, and transgenic technology should bring a new era to the bacterial blight resistance hybrid rice breeding program. This review also briefly discusses and deliberates on issues related to the broadening of bacterial blight resistance, and suitable utilization of resistance genes, alternate planting of available resistance genes; and understands the virulent populations of the bacterial pathogen in China even in Asia.

关键词： hybrid rice bacterial blight xanthomonas oryzae pv. oryzae resistance breeding genetics resistance gene

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XA23 Is an Executor R Protein and Confers Broad-Spectrum Disease Resistance in Rice

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Molecular Plant 2015年第2期8卷 290-302页

作者： Chunlian Wang Xiaoping Zhang Yinglun Fan Ying Gao Qinlong Zhu Chongke Zheng Tengfei Qin Yanqiang Li Jinying Che Mingwei Zhang Bing Yang Yaoguang Liu Kaijun Zhao National Key Facility for Crop Gene Resources and Genetic Improvement (NFCRI) Institute of Crop Science Chinese Academy of Agriculture Sciences (CAAS)Beijing 100081 China State Key Laboratory for Conservation and Utilization of Subtropical Agro-Bioresources College of Life Sciences South China Agricultural UniversityGuangzhou 510642 China Department of Genetics Development and Cell Biology Iowa State University Ames IA 50011 USA These authors contributed equally to the article. Present address: Liaocheng University Liaocheng 252059 Shandong China

The majority of plant disease resistance （R） genes encode proteins that share common structural features. However, the transcription activator-like effector （TALE）-associated executor type R genes show no considerable sequence homology to any known R genes. We adopted a map-based cloning approach and TALE-based technology to isolate and characterize Xa23, a new executor R gene derived from wild rice （Oryza rufipogon） that confers an extremely broad spectrum of resistance to bacterial blight caused by xanthomonas oryzae pv. oryzae （Xoo）. Xa23 encodes a 113 amino acid protein that shares 50% identity with the known executor R protein XA10. The predicted transmembrane helices in XA23 also overlap with those of XA10. Unlike XalO, however, Xa23 transcription is specifically activated by AvrXa23, a TALE present in all examined Xoo field isolates. Moreover, the susceptible xa23 allele has an identical open reading frame of Xa23 but differs in promoter region by lacking the TALE binding element （EBE） for AvrXa23. XA23 can trigger a strong hypersensitive response in rice, tobacco, and tomato. Our results provide the first evidence that plant genomes have an executor R gene family of which members execute their function and spectrum of disease resistance by recognizing the cognate TALEs in the pathogen.

关键词： Xa23 executor R gene TAL effector xanthomonas oryzae pv. oryzae AvrXa23 rice

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An XA21-Associated Kinase （OsSERK2） Regulates Immunity Mediated by the XA21 and XA3 Immune Receptors

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Molecular Plant 2014年第5期7卷 874-892页

作者： Xuewei Chena Shimin Zuo Benjamin Schwessinger Mawsheng Chern Patrick E. Canlas Deling Ruan Xiaogang Zhou Jing Wang Arsalan Daudi Christopher J. Petzold Joshua L. Heazlewood Pamela C. Ronald Department of Plant Pathology and the Genome Center University of California Davis CA 95616 USA Rice Research Institute Sichuan Agricultural University Chengdu Sichuan 611130 China Joint Bioenergy Institute Emeryville CA 94710 USA Key Laboratory of Crop Genetics and Physiology of Jiangsu Province Key Laboratory of Plant Functional Genomics of the Ministry of Education College of Agriculture Yangzhou University Yangzhou 225009 China

The rice XA21 immune receptor kinase and the structurally related XA3 receptor confer immunity to xanthomonas oryzae pv. oryzae （Xoo）, the causal agent of bacterial leaf blight. Here we report the isolation of OsSERK2 （rice somatic embryogenesis receptor kinase 2） and demonstrate that OsSERK2 positively regulates immunity mediated by XA21 and XA3 as well as the rice immune receptor FLS2 （OsFLS2）. Rice plants silenced for OsSerk2 display altered morphology and reduced sensitivity to the hormone brassinolide. OsSERK2 interacts with the intracellular domains of each immune receptor in the yeast two-hybrid system in a kinase activity-dependent manner. OsSERK2 undergoes bidi- rectional transphosphorylation with XA21 in vitro and forms a constitutive complex with XA21 in vivo. These results demonstrate an essential role for OsSERK2 in the function of three rice immune receptors and suggest that direct interaction with the rice immune receptors is critical for their function. Taken together, our findings suggest that the mechanism of OsSERK2-meditated regulation of rice XA21, XA3, and FLS2 differs from that of AtSERK3/BAK1-mediated regulation of Arabidopsis FLS2 and EFR.

关键词： immune receptor kinases somatic embryogenesis receptor kinase (SERK) immunity xanthomonas oryzae pv. oryzae rice.

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