The recombinant pea defensin Drr230a is active against impacting soybean and cotton pathogenic fungi from the genera Fusarium, Colletotrichum and Phakopsora
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- 作者：Ariane Ferreira Lacerda ; Rafael Perseghini Del Sarto ; Marilia Santos Silva…
- 关键词：Defensin ; Pisum sativum ; Pichia pastoris ; Fusarium tucumaniae ; Colletotrichum gossypii var. cephalosporioides ; Phakopsora pachyrhizi
- 刊名：3 Biotech
- 出版年：2016
- 出版时间：December 2016
- 年：2016
- 卷：6
- 期：1
- 全文大小：3,579 KB
- 参考文献：Abdallah NA, Shah D, Abbas D, Madkour M (2010) Stable integration and expression of a plant defensin in tomato confers resistance to Fusarium wilt. GM Crops 1–5:344–350CrossRef
Ahuja I, Kissen R, Bones AM (2012) Phytoalexins in defense against pathogens. Trends Plant Sci 2:73–90CrossRef
Almeida MS, Cabral KS et al (2001) cDNA cloning and heterologous expression of functional cysteine-rich antifungal protein Psd1 in the yeast Pichia pastoris. Arch Biochem Biophys 395:199–207CrossRef
Anuradha S, Divya K, Jami SK, Kirti PB (2008) Transgenic tobacco and peanut plants expressing a mustard defensin show resistance to fungal pathogens. Plant Cell Rep 27:1777–1786CrossRef
Aoki T, O’donell K, Homma Y, Lattanzi AR (2003) Sudden-death syndrome of soybean is caused by two morphologically and phylogenetically distinct species within the Fusarium solani species complex—F. virguliforme in North America and F. tucumaniae in South America. Mycologia 95:660–684CrossRef
Bednarek P (2012) Chemical warfare or modulators of defense responses-the function of secondary metabolites in plant immunity. Curr Opin Plant Biol 15:407–414CrossRef
Bloch C Jr, Richardson M (1991) A new family of small (5 kDa) protein inhibitors of insect α-amylases from seeds or sorghum (Sorghum bicolor (L) Munch) have sequence homologies with wheat γ-purothionins. FEBS Lett 279:101–104CrossRef
Cabral KMS, Almeida MS, Valente AP, Almeida FCL, Kurtenbach E (2003) Production of the active antifungal Pisum sativum defensin 1 (Psd1) in Pichia pastoris: overcoming the inefficiency of the STE13 protease. Protein Expr Purif 31:115–122CrossRef
Carvalho AO, Gomes VM (2009) Plant defensins—prospects for the biological functions and biotechnological properties. Peptides 30:1007–1020CrossRef
Chan YL, Prasad V et al (2005) Transgenic tomato plants expressing an Arabidopsis thionin (Thi2.1) driven by fruit-inactive promoter battle against phytopathogenic attack. Planta 221:386–393CrossRef
Chen KC, Lin CY et al (2002) Cloning and characterization of a cDNA encoding an antimicrobial protein from mung bean seeds. Bot Bull Acad Sin 43:251–259
Chen SC, Liu AR, Zou ZR (2006) Overexpression of glucanase gene and defensin gene in transgenic tomato enhances resistance to Ralstonia solanacearum. Russia J Plant Physiol 53:756–763CrossRef
Choy MS, Kim YH et al (2009) Expression of Br D1, a plant defensin from Brassica rape, confers resistance against brown plant hopper (Nilaparvata lugens) in transgenic rices. Mol Cells 28:131–137CrossRef
De Bondt A, Zaman S et al (1999) Genetic transformation of apple (Malus pumila Mill.) for increased fungal resistance: in vitro antifungal activity in protein extracts of transgenic apple expressing Rs-AFP2 or AceAMP1. Acta Hort 484:565–570
de Zélicourt A, Letousey P et al (2007) Ha-DEF1, a sunflower defensin, induces cell death in Orobanche parasitic plants. Planta 226:591–600CrossRef
Demain AL, Vaishnav P (2009) Production of recombinant proteins by microbes and higher organisms. Biotechnol Adv 27:297–306CrossRef
Fickers P (2014) Pichia pastoris: a workhorse for recombinant protein production. Curr Res Microbiol Biotechnol 2:354–363
Gao AG, Hakimi SM et al (2000) Fungal pathogen protection in potato by expression of a plant defensin peptide. Nat Biotechnol 18:1307–1310CrossRef
Godoy CV, Koga LJ, Canteri MG (2006) Diagrammatic scale for assessment of soybean rust severity. Fitopatol Bras 31:63–68CrossRef
Habib H, Fazili KM (2007) Plant protease inhibitors: a defense strategy in plants. Biotechnol Mol Biol Rev 2:68–85
Horbach R, Navarro-Quesada AR, Deising HB (2011) When and how to kill a plant cell: infection strategies of plant pathogenic fungi. J Plant Physiol 168:51–62CrossRef
Jha S, Chattoo BB (2010) Expression of a plant defensin in rice confers resistance to fungal phytopathogens. Transg Res 19:373–384CrossRef
Jrchen J, Hongchen G, Chinghsu H, Shingli S, Sanchen C (2004) Cloning and functional expression of a mungbean defensin VrD1 in Pichia pastoris. Agric Food Chem 52:2256–2261CrossRef
Kant P, Liu W, Peter PK (2009) PDC1, a corn defensin peptide expressed in Escherichia coli and Pichia pastoris inhibits growth of Fusarium graminearum. Peptides 30:1593–1599CrossRef
Kovalskaya N, Hammond RW (2009) Expression and functional characterization of the plant antimicrobial snakin-1 and defensin recombinant proteins. Protein Expr Purif 63:12–17CrossRef
Kristensen AK, Brunstedt J et al (1999) Processing, disulfide pattern, and biological activity of a sugar beet defensin, AX2, expressed in Pichia pastoris. Protein Expr Purif 16:377–387CrossRef
Lai FM, DeLong C, Mei K, Wignes T, Fobert PR (2002) Analysis of the DRR230 family of pea defensins: gene expression pattern and evidence of broad host-range antifungal activity. Plant Sci 163:855–864CrossRef
Lay FT, Anderson MA (2005) Defensins—components of the innate immune system in plants. Curr Protein Pept 6:85–101CrossRef
Lobo DS, Pereira IB et al (2007) Antifungal Pisum sativum defensin 1 interacts with Neurospora crassa cyclin F related to the cell cycle. Biochemistry 46:987–996CrossRef
Melo FR, Rigden DJ, Franco OL, Mello LV, Ary MB, Grossi de Sa MF, Bloch C Jr (2002) Inhibition of trypsin by cowpea thionin: characterization, molecular modeling, and docking. Proteins 48:311–319CrossRef
Mendez E, Moreno A et al (1990) Primary structure and inhibition of protein synthesis in eukaryotic cell-free system of a novel thionin, gamma-hordothionin, from barley endosperm. Eur J Biochem 194:533–539CrossRef
Mendez E, Rocher A et al (1996) Primary structure of omega-hordothionin, a member of a novel family of thionins from barley endosperm, and its inhibition of protein synthesis in eukaryotic and prokaryotic cell-free systems. Eur J Biochem 239:67–73CrossRef
Muthamilarasan M, Prasad M (2013) Plant innate immunity: an updated insight into defense mechanism. J Biosci 38:433–449CrossRef
Parashina EV, Serdobinskii LA et al (2000) Genetic engineering of oilseed rape and tomato plants expressing a radish defensin gene. Russ J Plant Physiol 47:417–423
Park HC, Kang YH et al (2002) Characterization of a stamen-specific cDNA encoding a novel plant defensin in Chinese cabbage. Plant Mol Biol 50:57–68CrossRef
Pelegrini PB, Lay FT, Murad AM, Anderson MA, Franco OL (2008) Novel insights on the mechanism of action of α-amylase inhibitors from the plant defensin family. Proteins 73:719–729CrossRef
Portieles R, Ayra C et al (2010) NmDef02, a novel antimicrobial gene isolated from Nicotiana megalosiphon confers high-level pathogen resistance under greenhouse and field conditions. Plant Biotechnol 8:678–690CrossRef
Rojas CM, Senthil-Kumar M, Tzin V, Mysore KS (2014) Regulation of primary plant metabolism during plant–pathogen interaction and its contribution to plant defense. Front Plant Sci 10:5–17
Schagger H, Von Jagow G (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal Biochem 166:368–379CrossRef
Seo HH, Park S et al (2014) Overexpression of a defensin enhances resistance to a fruit-specific anthracnose fungus in pepper. PLoS ONE 9:e97936CrossRef
Song X, Wang J, Wu F, Li X, Teng M, Gong W (2005) cDNA cloning, functional expression and antifungal activities of a dimeric plant defensin SPE10 from Pachyrhizus erosus seeds. Plant Mol Biol 57:13–20CrossRef
Spelbrink RG, Dilmac N et al (2004) Differential antifungal and calcium channel-blocking activity among structurally related plant defensins. Plant Physiol 135:2055–2067CrossRef
Towbin H, Staehelin T, Gordon J (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets—procedure and some applications. PNAS 76:4350–4354CrossRef
Vassileva A, Chugh DA (2001) Effect of copy number on the expression levels of hepatitis B surface antigen in the methylotrophic yeast Pichia pastoris. Protein Expr Purif 21:71–80CrossRef
Vijayan S, Guruprasad L, Kirti PB (2008) Prokaryotic expression of a constitutively expressed Tephrosia villosa defensin and its potent antifungal activity. Appl Microbiol Biotechnol 80:1023–1032
Wang YP, Fristensky B (2001) Transgenic canola lines expressing pea defense gene DRR206 have resistance to aggressive blackleg isolates and to Rhizoctonia solani. Mol Breed 8:263–271CrossRef
Wang Y, Nowak G (1999) Constitutive expression of pea defense gene DRR206 confers resistance to blackleg (Leptosphaeria maculans) disease in transgenic canola (Brassica napus). Mol Plant Microbe Interact 12:410–418CrossRef
Wijaya R, Neumann GM, Condron R, Hughes AB, Polya GM (2000) Defense proteins from seed of Cassia fistula include a lipid transfer protein homologue and a protease inhibitor plant defensin. Plant Sci 159:243–255CrossRef
Wong JH, Ng TB (2005) Sesquin, a potent defensin-like antimicrobial peptide from ground beans with inhibitory activities toward tumor cells and HIV-1 reverse transcriptase. Peptides 26:1120–1126CrossRef
Ye XY, Ng TB (2001) Peptides from pinto bean and red bean with sequence homology to cowpea 10-kda protein precursor exhibit antifungal, mitogenic, and HIV-1 reverse transcriptase-inhibitory activities. Biochem Biophys Res Commun 285:424–429CrossRef
Zhu YJ, Agbayani R, Moore PH (2007) Ectopic expression of Dahlia merckii defensin DmAMP1 improves papaya resistance to Phytophthora palmivora by reducing pathogen vigor. Planta 226:87–97CrossRef - 作者单位：Ariane Ferreira Lacerda (1) (2)
Rafael Perseghini Del Sarto (1) (3)
Marilia Santos Silva (1)
Erico Augusto Rosas de Vasconcelos (1)
Roberta Ramos Coelho (1)
Vanessa Olinto dos Santos (1) (2)
Claudia Vieira Godoy (4)
Claudine Dinali Santos Seixas (4)
Maria Cristina Mattar da Silva (1)
Maria Fatima Grossi-de-Sa (1) (5)
1. Embrapa Recursos Genéticos e Biotecnologia, PqEB, Avenida W5 Norte (Final), PO 02372, Brasília, DF, 70770-917, Brazil
2. Universidade Federal do Rio Grande do Norte, Avenida Senador Salgado Filho 3000, Lagoa Nova, Natal, RN, 59078-970, Brazil
3. Centro Universitário do Distrito Federal, SEP/SUL EQ 704/ A12 904 Conjunto A, Brasília, DF, 70390-045, Brazil
4. Embrapa Soja, Rodovia Carlos João Strass, Distrito de Warta, A10 PO Box 231, Distrito de Warta, PR, 86001-970, Brazil
5. Universidade Católica de Brasília, SGAN 916 Módulo B Avenida W5, Brasília, DF, 70790-160, Brazil - 刊物主题：Biotechnology; Agriculture; Cancer Research; Bioinformatics; Stem Cells; Biomaterials;
- 出版者：Springer Berlin Heidelberg
- ISSN：2190-5738
文摘
Plant defensins are antifungal peptides produced by the innate immune system plants developed to circumvent fungal infection. The defensin Drr230a, originally isolated from pea, has been previously shown to be active against various entomopathogenic and phytopathogenic fungi. In the present study, the activity of a yeast-expressed recombinant Drr230a protein (rDrr230a) was tested against impacting soybean and cotton fungi. First, the gene was subcloned into the yeast expression vector pPICZαA and expressed in Pichia pastoris. Resulting rDrr230a exhibited in vitro activity against fungal growth and spore germination of Fusarium tucumaniae, which causes soybean sudden death syndrome, and against Colletotrichum gossypii var. cephalosporioides, which causes cotton ramulosis. The rDrr230a IC50 corresponding to inhibition of fungal growth of F. tucumaniae and C. gossypii var. cephalosporioides was 7.67 and 0.84 µM, respectively, demonstrating moderate activity against F. tucumaniae and high potency against C. gossypii var. cephalosporioides. Additionally, rDrr230a at 25 ng/µl (3.83 µM) resulted in 100 % inhibition of spore germination of both fungi, demonstrating that rDrr230a affects fungal development since spore germination. Moreover, rDrr230a at 3 µg/µl (460.12 µM) inhibited 100 % of in vitro spore germination of the obligatory biotrophic fungus Phakopsora pachyrhizi, which causes Asian soybean rust. Interestingly, rDrr230a substantially decreased the severity of Asian rust, as demonstrated by in planta assay. To our knowledge, this is the first report of a plant defensin active against an obligatory biotrophic phytopathogenic fungus. Results revealed the potential of rDrr230a as a candidate to be used in plant genetic engineering to control relevant cotton and soybean fungal diseases. Keywords Defensin Pisum sativum Pichia pastoris Fusarium tucumaniae Colletotrichum gossypii var. cephalosporioides Phakopsora pachyrhizi