|
|
|
| Bioinformatic Analysis and Promoter Identification of clt-1 Gene in Curvularia Lunata |
| NI Xuan1,2, GAO Jin-xin1,2, YU Chuan-jin1,2, LIU Tong1,2, LI Ya-qian1,2, CHEN Jie1,2 |
1. Key Laboratory of Urban Agriculture(South, School of Agriculture and Biology, Shanghai Jiaotong University, Shanghai 200240, China;
2. State Key Laboratory of Microbial Metabolism, Shanghai Jiaotong University, Shanghai 200240, China |
|
|
|
Abstract The clt-1 gene within Curvularia lunata genomic sequence was screened through Agrobacterium-mediated transformation (ATMT) method. Bioinformatic analysis showed that the molecular weight of the protein encoded by clt-1 was 81.984 8kDa, the oretical isoelectric point pI was 8.62, and the instability index was 55.08. CLT-1 was a hydrophilic protein, containing one transmembrane helix topologies. At subcellular level, CLT-1 protein located in the nucleus. More than one of the serine, threonine and tyrosine kinase phosphorylation sites in CLT-1 amino acids sequence were found. In the aspect of its function, CLT-1 protein contained a typical conserved BTB function domain, which may regulate the expression of some functional protein. The expression vector pC1300th-Pclt1-GFP was constructed for detecting its promoter functional activity by the fusion of clt-1 promoter fragment to the reporter GFP gene. Transformation of plasmid pC1300th-Pclt1-GFP was conducted by ATMT method of the spores of C. lunata wild-type CX-3. The putative transformants were obtained from selective regeneration medium and confirmed by PCR detection and green fluorescence analysis. The results indicated that GFP gene was integrated into the genome of CX-3 strain, and the clt-1 promoter could drive the GFP gene expression. Strong green fluorescence activity was detectable in the mycilia and spores of transformants under the confocal laser scanning microscope.
|
|
Received: 18 September 2016
Published: 25 March 2017
|
|
|
|
|
| [1] 吕国忠,陈捷,白金铠,等. 我国玉米病害发生现状及防治措施. 植物保护,2007,23(4):20-21. Lv G Z, Chen J, Bai J K, et al. The occurrence status and control measures of maize disease in China. Plant Protection, 1997, 23(4):20-21.
[2] 李富华,王玉涛,潘开文.玉米弯孢叶斑病研究现状、问题与展望.植物保护,2004,30(6):5-10. Li F H, Wang Y T, Pan K W. Progresses, problems and prospects of maize Curvularia leaf spot disease.Plant Protection, 2004, 30(6):5-10.
[3] 赵君,王国英,胡剑,等. 玉米弯孢菌叶斑病抗性的ADAA遗传模型的分析. 作物学报,2002,28(1):127-132. Zhao J, Wang G Y, Hu J, et al. Genetic analysis of maize resistance to Curvularia leaf spot by ADAA model. Acta Agronomica Sinica. 2002, 28(1):127-130.
[4] 傅俊范,李海春,白元俊,等. 玉米弯孢菌叶斑病传播梯度模型.植物病理学报,2003,33(5):456-461. Fu J F, Li H C, Bai Y J, et al. Spread gradient model of leaf spot disease (Curvularia lunata) in maize. Acta Phytopathologica Sinica, 2003, 33(5):456-461.
[5] 薛春生,肖淑芹,翟羽红,等. 玉米弯孢叶斑病菌致病类型分化研究. 植物病理学报,2008,38(1):6-12. Xue C S, Xiao S Q, Zhai Y H, et al. Pathogenicity differentiation of Curvularia lunata. Acta Phytopathologica Sinica, 2008, 38(1):6-12.
[6] Gao J X, Liu T, Chen J. Insertional mutagenesis and cloning of the gene required for the biosynthesis of the non-host specific toxin in Cochliobolus lunatus that causes maize leaf spot. Phytopathology, 2014, 104(4):332-339.
[7] Liu T, Liu L X, Jiang X, et al. Agrobacterium-mediated transformation as a useful tool for the molecular genetic study of the phytopathogen Curvularia lunata. Europe Journal of Plant Pathology, 2010, 126(3):363-371.
[8] Gao J X, Jing J, Liu T, et al. Identification of Clt-1-regulated proteins associated with the production of non-host-specific toxin and pathogenicity in Cochliobolus lunatus. Journal of Phytopathology, 2015, 163(1):33-41.
[9] Chen J S, Zheng W, Zheng S Q, et al. Rac1 is Required for pathogenicity and Chem1-dependent conidiogenesis in rice fungal pathogen Magnaporthe grisea. PLoS One, 2008, 4(11):1-16.
[10] Yi M, Park J H, Ahn J H, et al. MoSNF1 regulates sporulation and pathogenicity in the rice blast fungus Magnaporthe oryzae. Fungal Genetics and Biology, 2008, 45(8), 1172-1181.
[11] Steiner U, Oerke E C. Localized melanization of appressoria is required for pathogenicity of Venturia inaequalis. Phytopathology, 2008, 97(10):1222-1230. |
|
Viewed |
|
|
|
Full text
|
|
|
|
|
Abstract
|
|
|
|
|
Cited |
|
|
|
|
| |
Shared |
|
|
|
|
| |
Discussed |
|
|
|
|
