Antimicrobial activities of ant Ponericin W1 against plant pathogens in vitro and the disease resistance in its transgenic Arabidopsis
Received date: 2013-04-01
Revised date: 2013-05-08
Online published: 2013-08-25
The antimicrobial peptides (AMPs) exhibit a broad antimicrobial spectrum. The application of AMPs from non-plant organisms attracts considerable attention in plant disease resistance engineering. Ponericin W1, isolated from the venom of ant (Pachycondyla goeldii), shows antimicrobial activities against Gram-positive bacteria, Gram-negative bacteria and the budding yeast (Saccharomyces cerevisiae); however, it is not clear whether Ponericin W1 is effective against plant pathogens. The results of this study indicated synthesized Ponericin W1 inhibited mycelial growth of Magnaporthe oryzae and Botrytis cinerea, as well as hyphal growth and spore production of Fusarium graminearum. Besides, Ponericin W1 exhibited antibacterial activities against Pseudomonas syringae pv. tomato and Xanthomonas oryzae pv. oryzae. After codon optimization, Ponericin W1 gene was constructed into plant expression vector, and transformed into Arabidopsis thaliana by floral dip method. The Ponericin W1 was located in intercellular space of the transgenic plants as expected. Compared with the wild-type plants, there were ungerminated spores and less hyphal, conidia on the leaves of transgenic plants after innoculation with the powdery mildew fungus Golovinomyces cichoracearum. After innoculation with the pathogenic bac-terium Pseudomonas syringae pv. tomato, the baceria in the leaves of transgenic plants was significantly less than the wild-type plants, indicating that the transgenic plants displayed enhanced disease resistance to pathogens. These results demonstrate a potential use of Ponericin W1 in genetic engineering for broad-spectrum plant disease resistance.
Key words： Ponericin W1; transgenic; antimicrobial peptides; plant pathogen
CHEN Yong-Fang SUN Peng-Wei TANG Ding-Zhong . Antimicrobial activities of ant Ponericin W1 against plant pathogens in vitro and the disease resistance in its transgenic Arabidopsis[J]. Hereditas(Beijing), 2013 , 35(8) : 1023 -1029 . DOI: 10.3724/SP.J.1005.2013.01023
[1] Zasloff M. Antimicrobial peptides of multicellular organ-isms. Nature, 2002, 415(6870): 389-395.
[2] 金莉莉, 王秋雨. 蛙科两栖动物皮肤抗菌肽的分子多样性及功能. 遗传, 2008, 30(10): 1241-1248.
[3] Matsuzaki K. Why and how are peptide-lipid interactions utilized for self-defense? Magainins and tachyplesins as archetypes. Biochim Biophys Acta, 1999, 1462(1-2): 1-10.
[4] Yang L, Weiss TM, Lehrer RI, Huang HW. Crystallization of antimicrobial pores in membranes: magainin and pro-tegrin. Biophys J, 2000, 79(4): 2002-2009.
[5] Westerhoff HV, Juretic D, Hendler RW, Zasloff M. Magainins and the disruption of membrane-linked free-energy trans-duction. Proc Natl Acad Sci USA, 1989, 86(17): 6597-6601.
[6] Melo MN, Castanho MARB. The mechanism of action of antimicrobial peptides: lipid vesicles vs. bacteria. Front Immunol, 2012, 3: 236.
[7] Odintsova T, Egorov T. Plant antimicrobial peptides. In: Irving HR, Gehring C, eds. Plant Signaling Peptides. Ber-lin: Springer, 2012, 16: 107-133.
[8] Montesinos E. Antimicrobial peptides and plant disease control. FEMS Microb Lett, 2007, 270(1): 1-11.
[9] 宫霞, 乐国伟, 施用晖. 昆虫抗菌肽及其基因工程研究进展. 遗传, 2002, 24(5): 591-594.
[10] Orivel J, Redeker V, Le Caer JP, Krier F, Revol-Junelles AM, Longeon A, Chaffotte A, Dejean A, Rossier J. Pon-ericins, new antibacterial and insecticidal peptides from the venom of the ant Pachycondyla goeldii. J Biol Chem, 2001, 276(21): 17823-17829.
[11] Clough SJ, Bent AF. Floral dip: a simplified method for Agrobacterium-mediated transformation of Arabidopsis thaliana. Plant J, 1998, 16(6): 735-743.
[12] Fan J, Crooks C, Lamb C. High-throughput quantitative luminescence assay of the growth in planta of Pseudomonas syringae chromosomally tagged with Photorhabdus luminescens luxCDABE. Plant J, 2008, 53(2): 393-399.
[13] Nie HZ, Wu YY, Yao CP, Tang DZ. Suppression of edr2-mediated powdery mildew resistance, cell death and ethylene-induced senescence by mutations in ALD1 in Arabidopsis. J Genet Genomics, 2011, 38(4): 137-148.
[14] Hightower R, Baden C, Penzes E, Dunsmuir P. The expression of cecropin peptide in transgenic tobacco does not confer resistance to Pseudomonas syringae pv. tabaci. Plant Cell Rep, 1994, 13(5): 295-299.
[15] Owens LD, Heutte TM. A single amino acid substitution in the antimicrobial defense protein cecropin B is associated with diminished degradation by leaf intercellular fluid. Mol Plant Microbe Interact, 1997, 10(4): 525-528.
[16] Oard S, Enright FM. Expression of the antimicrobial peptides in plants to control phytopathogenic bacteria and fungi. Plant Cell Rep, 2006, 25(6): 561-572.
[17] Lee SB, Li BC, Jin SX, Daniell H. Expression and char-acterization of antimicrobial peptides Retrocyclin-101 and Protegrin-1 in chloroplasts to control viral and bacterial infections. Plant Biotechnol J, 2011, 9(1): 100-115.
[18] 贾士荣, 屈贤铭, 冯兰香, 唐惕, 唐益雄, 刘坤, 赵艳丽, 白永延, 蔡敏莺. 转抗菌肽基因提高马铃薯对青枯病的抗性. 中国农业科学, 1998, 31(3): 5-12.
[19] 李春风, 张兴政, 张洁莉, 韩阳, 宋书宏. 根癌农杆菌介导抗菌肽基因转化大豆的研究. 中国油料作物学报, 2012, 34(4): 348-352.
[20] 黄大年, 朱冰, 杨炜, 薛锐, 肖晗, 田文忠, 李良才, 戴顺洪. 抗菌肽B基因导入水稻及转基因植株的鉴定. 中国科学(C辑: 生命科学), 1997, 27(1): 55-62.
[21] 杨凯, 韩伟, 温莹, 刘丹丹, 薛春蕾, 逯晓萍. 转Bt和抗菌肽融合基因油菜植株的获得与鉴定研究. 华北农学报, 2011, 26(2): 49-54.
[22] 田长恩, 王正询, 陈韬, 周玉萍, 黄自然, 黄亚东. 抗菌肽D基因导入番茄及转基因植株的鉴定. 遗传, 2000, 22(2): 86-89.
/
| 〈 |
|
〉 |
www.chinagene.cn
备案号：京ICP备09063187号
