Enhanced Resistance to Phytopathogenic Bacteria in Transgenic Tobacco Plants with Synthetic Gene of Antimicrobial Peptide Cecropin P1

Abstract

Plasmids with a synthetic gene of the mammalian antimicrobial peptide cecropin P1 ( cecP1 ) controlled by the constitutive promoter 35S RNA of cauliflower mosaic virus were constructed. Agrobacterial transformation of tobacco plants was conducted using the obtained recombinant binary vector. The presence of gene cecP1 in the plant genome was confirmed by PCR. The expression of gene cecP1 in transgenic plants was shown by Northern blot analysis. The obtained transgenic plants exhibit enhanced resistance to phytopathogenic bacteria Pseudomonas syringae, P. marginata, and Erwinia carotovora. The ability of transgenic plants to express cecropin P1 was transmitted to the progeny. F 0 and F 1 plants had the normal phenotype (except for a changed coloration of flowers) and retained the ability to produce normal viable seeds upon self-pollination. Lines of F 1 plants with Mendelian segregation of transgenic traits were selected.

Insect peptide metchnikowin confers on barley a selective capacity for resistance to fungal ascomycetes pathogens

“…There are a number of reports in which antimicrobial peptides of insect ( Osusky et al , 2000 ; Langen et al , 2006 ), frog ( Chakraborti et al , 2003 ; Yevtushenko and Misra, 2007 ) or human ( Zakharchenko et al , 2005 ; Aerts et al , 2007 ) have been used to render the transformed plants more resistant to fungal pathogens. Beside the evaluation of Mtk effects on the development of pathogenic fungi, its possible adverse impact on the beneficial root endophyte Piriformospora indica ( Waller et al , 2005 ) has been investigated, too.…”

Novel expression system for enhanced synthesis of antimicrobial peptide cecropin P1 in plants

“…In order to enhance the synthesis of CP1 in plants, we used vector pPCV91 [10] containing superpromoter CaMV 35S with four enhancer domains [27] and nontranslated Ω leader sequence of tobacco mosaic virus in the present work [28]. Expression of gene СР1 under the control of these genetic elements allowed us to intensify the syn thesis CP1 to 0.2% of total level of soluble leaf protein as compared with expression of the same gene under the control of promoter CaMV 35S with one enhancer domain (0.002%) [7]. The transgenic plants obtained possessed increased resistance to phytopathogen strains of E. carotovora and S. sclerotiorum as com pared with the control ones.…”

“…Here, stable expres sion of cecropin at the level of 0.002% of total soluble leaf protein was noted [7,8]. Enhancement of expres sion of antimicrobial peptides in transgenic plants can considerably increase their tolerance to phytopatho genic microorganisms and should allow us to consider these plants as producers of such compounds.The pur pose of the present work was to enhance the synthesis of cecropin P1 (CP1) [9] in plants using novel expression system where gene CP1 is under the control of "super promoter" CaMV 35S containing 4 enhancer sequences CaMV 35S and nontranslated leader sequence Ω RNA of tobacco mosaic virus [10].…”

“…Radius of clear area around the well served as an indicator of bacteria growth inhibition extent. Quantitative evalu ation of CP1 content in the extracts of transgenic plants was conducted by means of comparing with standard concentrations of synthetic CP1 [7].…”

Antimicrobial peptides (AMPs): A promising class of antimicrobial compounds

“…syringae , Pseudomonas marginata and E . carotovora (Zakharchenko et al, 2005). In several studies focused on MSI‐99, an analogue of magainin was expressed in different plants, such as tobacco, banana, tomato and grapevine, which demonstrated effective protection against a variety of fungal and bacterial phytopathogens (Alan et al, 2004; Chakrabarti et al, 2003; Vidal et al, 2006).…”