1.Advances in albicidin.
Lilan CHEN ; Haibin HUANG ; Runtian BIAN ; Zuhu DENG ; Sanji GAO ; Huili ZHANG
Chinese Journal of Biotechnology 2022;38(8):2738-2753
Xanthomonas albilineans (Ashby) Downson is a quarantine pest for importing plants to China that causes leaf scald bacterial disease on sugarcane. X. albilineans produces a potent phytotoxin/antibiotic called albicidin. As a pathogenic factor, albicidin causes typical white leaf stripes by inhibiting plastid DNA gyrase and disturbing chloroplast differentiation. Meanwhile, the antibacterial activity of albicidin gives X. albilineans a competitive advantage against rival bacteria during their colonization. Furthermore, albicidin has a rapid bactericidal activity against a variety of Gram-positive and Gram-negative pathogenic bacteria of human species at nanomolar concentrations, making it a potential antimicrobial drug for clinical application. This article reviews the advances of albicidin from the aspects of its molecular structure, traditional extraction methods, mechanism of action, biosynthetic genes and processes, chemical synthesis method and improvement, in order to provide insights into the prevention and treatment of the sugarcane leaf scald disease, and the development of new antibiotics.
Anti-Bacterial Agents/pharmacology*
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China
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Humans
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Organic Chemicals
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Xanthomonas/genetics*
2.The role of iron-uptake factor PiuB in pathogenicity of soybean pathogen Xanthomonas axonopodis pv. glycines.
Ruyi SU ; Luojia JIN ; Jiangling XU ; Huiya GENG ; Xiao CHEN ; Siyi LIN ; Wei GUO ; Zhiyuan JI
Chinese Journal of Biotechnology 2024;40(1):177-189
Iron is an essential element for living organisms that plays critical roles in the process of bacterial growth and metabolism. However, it remains to be elucidated whether piuB encoding iron-uptake factor is involved in iron uptake and pathogenicity of Xanthomonas axonopodis pv. glycines (Xag). To investigate the function of piuB, we firstly generated a piuB deletion mutant (ΔpiuB) by homologous recombination. Compared with the wild-type, the piuB mutant exhibited significantly reduced growth and virulence in host soybean. The mutant displayed markedly increased siderophore secretory volume, and its sensitivity to Fe3+, Cu2+, Zn2+ and Mn2+ was significantly enhanced. Additionally, the H2O2 resistance, exopolysaccharide yield, biofilm formation, and cell mobility of ΔpiuB were significantly diminished compared to that of the wild-type. The addition of exogenous Fe3+ cannot effectively restore the above characteristics of ΔpiuB. However, expressing piuB in trans rescued the properties lost by ΔpiuB to the levels in the wild-type. Taken together, our results demonstrated that PiuB is a potential factor for Xag to assimilate Fe3+, and is necessary for Xag to be pathogenic in host soybean.
Iron
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Glycine max
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Virulence
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Xanthomonas axonopodis/genetics*
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Hydrogen Peroxide
3.The gene wxcA of Xanthomonas campestris pv. campestris 8004 strain involved in EPS yield.
Guang-Tao LU ; Ji-Liang TANG ; Guang-Ning WEI ; Yong-Qiang HE ; Bao-Shan CHEN
Chinese Journal of Biotechnology 2004;20(4):477-483
Xanthomonas campestris pv. campestris (Xcc), the pathogenic agent of black rot disease in cruciferous plants, produces large amount of extracellular polysaccharide (EPS), which has found wide applications in industry. For the great commercial value of the xanthan gum, many of the genes involved in EPS biosynthesis have been cloned and the mechanism of EPS biosynthesis also has been studied. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5 gusA5, and a number of EPS-defective mutants were isolated in our previous work. The Tn5 gusA5 inserted sites of these mutants were located by using thermal asymmetric interlaced PCR, and results showed that two EPS-defective mutants were insertion mutants of the gene wxcA which involved in lipopolysaccharide (LPS) biosynthesis. The gene wxcA involved in lipopolysaccharide biosynthesis but dose not extracellular polysaccharide in others' report. wxcA::Tn5 gusA5 mutant 021C12, the polar mutant, was complemented with recombinant plasmid pLATC8570 harboring an intact wxcA gene in this work, but the yield of EPS of the wxcA::Tn5 gusA5 mutant was not restored. In order to identify the function of wxcA gene of Xcc 8004 strain, the gene wxcA was deleted by gene replacement strategy, and the no-polar mutant of wxcA was obtained. DeltawxcA mutant strain, named Xcc 8570, was confirmed by using both PCR and southern analysis. Beside the LPS biosynthesis of deltawxcA mutant was affected, The EPS yield of deltawxcA mutant strain reduced by 50% as compared with the wild-type strain 8004. DeltawxcA mutant could be complemented in trans with the intact wxcA gene, and the EPS yield of the mutant was restored. The combined data showed that wxcA gene not only involved in LPS biosynthesis but also EPS yield in Xcc 8004 strain.
Cell Proliferation
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Genes, Bacterial
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physiology
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Lipopolysaccharides
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biosynthesis
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Mutation
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Polysaccharides, Bacterial
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biosynthesis
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Xanthomonas campestris
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genetics
4.Introduction of a non-host gene Rxo1 cloned from maize resistant to rice bacterial leaf streak into rice varieties.
Xue-Wen XIE ; Jing YU ; Jian-Long XU ; Yong-Li ZHOU ; Zhi-Kang LI
Chinese Journal of Biotechnology 2007;23(4):607-611
Rice bacterial leaf streak,caused by Xanthomonas oryzae pv. oryzicola is a destructive bacterial disease in China. Single-gene resistance to X. oryzae pv. oryzicola has not been found in rice germplasm. A cloned non-host gene from maize with resistance to bacterial leaf streak, Rxo1, was transferred into four Chinese rice varieties through an Agrobacterium-mediated system, including Zhonghua11, 9804, C418 and Minghui86. PCR and Southern analysis of the transgenic plants revealed the integration of the Rxo1 gene into the rice genomes. The integrated Rxo1 was stably inherited, and segregated in a 3:1 (Resistance:Susceptible) ratio in the selfed T1 generations derived from some T0 plants, indicating that Rxo1 inherited as a dominate gene in rice. Transgenic T0 plants and PCR-positive T1 plants were resistant to X. oryzae pv. oryzicola on the basis of artificial inoculation.
Bacterial Proteins
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genetics
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metabolism
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Genes, Plant
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genetics
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Oryza
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genetics
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Plant Diseases
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genetics
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microbiology
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Plants, Genetically Modified
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genetics
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Rhizobium
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genetics
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Transformation, Genetic
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Xanthomonas
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genetics
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Zea mays
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genetics
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microbiology
5.Generation of selectable marker-free and vector backbone sequence-free Xa21 transgenic rice.
Zhi-Hui XIA ; Xiao-Bing LI ; Cai-Yan CHEN ; Hai-Kuo FAN ; Guang-Huai JIANG ; Li-Huang ZHU ; Wen-Xue ZHAI
Chinese Journal of Biotechnology 2006;22(2):204-210
The dominant gene Xa21 with broad-spectrum and high resistance to Xanthomonas oryzae pv. oryzae (Xoo) was transferred into C418, an important restorer line of japonica hybrid rice in China using double right-border (DRB) T-DNA binary vector through Agrobacterium-mediated transformation. 17 transgenic lines were Xa21-positive with high resistance to the race P6 of Xoo through PCR analysis and resistance identification, among the total 27 independent primary transformants (T0) obtained. The subsequent analysis of the T1 progenies of these 17 T0 lines through PCR-assisted selection and resistance investigation showed that four Xa21 transgenic T0 lines could produce selectable marker-free (SMF) progenies. The frequency of primary transformants producing SMF progenies was 15%. In addition, PCR analysis also revealed these SMF progenies did not contain vector backbone sequence, and they were named as SMF and vector backbone sequence-free (SMF-VBSF) Xa21 transgenic plants. The further molecular and phenotypic analysis of the T2 and T3 progenies testified the homozygous SMF-VBSF Xa21 transgenic plants were obtained with high resistance to Xoo.
DNA, Bacterial
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genetics
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Genetic Vectors
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Oryza
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genetics
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Plant Proteins
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genetics
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Plants, Genetically Modified
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genetics
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Protein-Serine-Threonine Kinases
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genetics
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Rhizobium
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genetics
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Transformation, Genetic
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Xanthomonas
6.Differentiation of xanthomonads causing the bacterial leaf spot of poinsettia in China from the pathotype strain of Xanthomonas axonopodis pv. poinsettiicola.
Bin LI ; Guan-lin XIE ; J SWINGS
Journal of Zhejiang University. Science. B 2005;6(6):451-453
In October 2003, a new bacterial disease with symptoms similar to those caused by Xanthomonas axonopodis pv. poinsettiicola was observed on poinsettia leaves at a flower nursery in Zhejiang Province of China. Three Xanthomonas strains were isolated from infected plants and classified as X. axonopodis. They were differentiated from the pathotype strain LMG849 of X. axonopodis pv. poinsettiicola causing bacterial leaf spot of poinsettia by comparison of pathogenicity, substrate utilization and BOX-PCR genomic fingerprints.
Cell Differentiation
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China
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Euphorbia
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microbiology
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Plant Diseases
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microbiology
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Plant Leaves
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microbiology
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Species Specificity
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Xanthomonas
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classification
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genetics
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isolation & purification
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pathogenicity
7.Accelerated improvement of bacterial blight resistance of 'Shuhui527' using molecular marker-assisted selection.
Ting-You HUANG ; Shi-Gui LI ; Yu-Ping WANG ; Han-Yun LI
Chinese Journal of Biotechnology 2003;19(2):153-157
'Shuhui527' is a promising restorer line bred by Rice Research Institute of Sichuan Agricultural University in recent years. However, this line is susceptible to Bacterial Blight (BB), which limits its use. The IRBB60, from the International Rice Research Institute (IRRI), contains dominant genes Xa21 and Xa4 conferring resistance to BB. The objective of this study is to improve the BB resistance of 'Shuhui527' by introgressing Xa21 and Xa4, the two broad-spectrum BB resistance genes, into 'Shuhui527' with IRBB60 as the donor, pTA248 and MP12, linking tightly with Xa21 and Xa4 respectively as DNA markers. BC1 F1 progenies of (Shuhui527 x IRBB60), containing Xa21 and Xa4 identified using PCR screening and with agronomic traits including plant type, grain type and days to heading etc similar to those of 'Shuhui527', were subsequently backcrossed to 'Shuhui527' and self-pollinated to generate BC2 F1 and BC1 F2 . The BC3 F1 and BC3 F2 were subsequently developed using the same approach. Among the 20 BC3 F2 plants, homozygous Xa21 and Xa4,10 plants were the most similar to 'Shuhui527' in the agronomic traits, and were screened using 120 pairs SSR and 100 pairs RAPD markers. Based on the results of the background screening and the performance of the agronomic traits, 5 plants were identified as improved-'Shuhui527' and designated as 527R-5, 527R-6, 527R-8, 527R-9 and 527R-10. The improved-' Shuhui527' lines expressed high resistance to Xanthomonas oryzae pv . oryzae (Xoo) stains C I - C VII, P1 and P6. The evaluation of the polymorphisms and selection accuracies of pTA248 and MP12 demonstrated that the polymorphisms of the two markers were obvious and co-dominant and the accuracies were more than 97% and 83% respectively, indicating the two markers are good for Xa21 and Xa4 in Molecular Marker-assisted Selection.
Genes, Plant
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genetics
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physiology
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Oryza
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genetics
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microbiology
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Plant Diseases
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genetics
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microbiology
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Plants, Genetically Modified
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genetics
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microbiology
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Polymerase Chain Reaction
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Xanthomonas
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pathogenicity
8.Synthesis of cefatrizine by recombinant alpha-amino acid ester hydrolase.
Jialin PAN ; Lu WANG ; Duanhua LI ; Lijuan YE
Chinese Journal of Biotechnology 2013;29(4):501-509
To explore the enzymatic route of cefatrizine synthesis, alpha-amino acid ester hydrolase (AEH) gene was cloned from the whole genome of Xanthomonas rubrillineans, and expressed heterologously in Escherichia coli BL21 (DE3). The effects of temperature, pH and substrates' molar ratio upon the transformation yield of cefatrizine by purified recombinant AEH were investigated. The monomer of AEH was determined as 70 kDa by SDS-PAGE. The optimal pH and temperature reaction were (6.0 +/- 0.1) and 36 degrees C for cefatrizine synthesis. The transformation yield was 64.3% under 36 degrees C, pH (6.0 +/- 0.1), when the concentrations of two substrates were about 30 mmol/L (7-ATTC) and 120 mmol/L (HPGM x HCl), respectively, and the enzyme consumption was 22 U/mL. The results pave the way for optimization of the industrial enzymatic synthesis of cefatrizine.
Carboxylic Ester Hydrolases
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biosynthesis
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genetics
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Catalysis
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Cefatrizine
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metabolism
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Kinetics
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Recombinant Proteins
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biosynthesis
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genetics
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Xanthomonas
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enzymology
9.Molecular recognition code between pathogenic bacterial TAL-effectors and host target genes: a review.
Yanqiang LI ; Chunlian WANG ; Kaijun ZHAO
Chinese Journal of Biotechnology 2011;27(8):1132-1141
As the pathogenic bacterial virulence and avirulence factors, transcription activator like (TAL) effectors of Xanthomonas can resulted in the host diseases or resistance responses. TAL effectors can specifically bind the target DNA of host plant with a novel protein-DNA binding pattern in which two amino acids recognize one nucleotide. The complexities of TAL-DNA binding have the feasibility in use of gene therapy through homologous recombination and site-specific mutation. By using the molecular recognition code between TAL-effectors and host target genes, we can exploit both the susceptible and resistance genes; broad spectrum resistance induced by multiple TAL effectors could also be manipulated. Deeper insight in the area of protein-DNA binding mechanism will benefit the application in the biomedical engineering and agricultural engineering. This article reviews the findings and functions of TAL effectors, the binding specificity and recognition code between TAL-effectors and host target genes. The possible applications and future prospects of the molecular recognition code have been discussed.
Base Sequence
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DNA, Plant
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metabolism
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Genes, Plant
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Genetic Code
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genetics
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Host-Pathogen Interactions
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Molecular Sequence Data
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Plant Diseases
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genetics
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prevention & control
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Transcriptional Activation
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Virulence Factors
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genetics
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metabolism
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Xanthomonas
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genetics
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pathogenicity
10.Identification and cloning of a novel gene involved in EPS biosynthesis of Xanthomonas campestris pv. campestris.
Guang-Tao LU ; Ji-Liang TANG ; Yong-Qiang HE ; Bao-Shan CHEN ; Dong-Jie TANG
Chinese Journal of Biotechnology 2003;19(6):661-667
Xanthomonas campestris pv. campestris ( Xcc), causative agent of the black rot disease of cruciferous crops worldwide, produces large amount of extracellular polysaccharide( EPS), which has found wide applications in industry. In order to clone genes involved in EPS biosynthesis, Xcc wild-type strain 8004 was mutagenized with transposon Tn5gus A5, and a number of EPS-defective mutants were isolated. The Tn5gusA5 insertion sites in the mutants were analyzed by using thermal asymmetric interlaced PCR(TAIL-PCR), and the corresponding genes were identified by homology blast to the completely sequenced genome of Xcc 8004 strain. A novel gene, waxE, identified from the EPS-defective mutant 151D09, was found to be disrupted by the insertion of Tn5gusA5 in the open reading frame(ORF) with genome coordinates 4478998bp to 4479819bp.This gene showed 52% similarity to the kdtX gene of Serratia marcescens and 50% to the waaE of Klebsiella pneumoniae at amino acid level, with characteristics of glycostransferase 2 family domain. In order to identify the function of waxE gene, waxE gene deletion mutant of Xcc 8004 was constructed by gene replacement strategy in which waxE gene of genome was replaced by kanamycin resistant gene kan. The waxE gene deletion mutant strain, named Xcc 8570, was confirmed by both PCR and southern analysis. The growth rate of the deletion mutant 8570 in rich medium was not affected, but the EPS yield reduced by 35% as compared with the wildtype strain 8004. The deletion mutant could be completmented in trans with plasmid pLATC8976 harboring an intact waxE gene, and the EPS yield of the mutant was restored. The combined data showed that waxE gene involved in EPS biosynthesis in Xcc.
Amino Acid Sequence
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Bacterial Proteins
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chemistry
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genetics
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metabolism
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Blotting, Southern
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Cloning, Molecular
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DNA Transposable Elements
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genetics
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Molecular Sequence Data
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Polymerase Chain Reaction
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Polysaccharides, Bacterial
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genetics
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metabolism
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Sequence Analysis, DNA
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Sequence Homology, Amino Acid
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Xanthomonas campestris
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genetics
;
metabolism