期刊
TRANSGENIC RESEARCH
卷 21, 期 1, 页码 23-37出版社
SPRINGER
DOI: 10.1007/s11248-011-9506-x
关键词
Tomato; Clavibacter michiganensis subsp michiganensis; Snakin-2; Extensin-like protein; T-DNA
类别
资金
- New York State Agricultural Experiment Station
- United States-Israel Binational Agricultural Research and Development Fund [IS-4047-07]
Two tomato proteins were evaluated by over-expression in transgenic tomato for their ability to confer resistance to Clavibacter michiganensis subsp. michiganensis (Cmm). Snakin-2 (SN2) is a cysteinerich peptide with broad-spectrum antimicrobial activity in vitro while extensin-like protein (ELP) is a major cell-wall hydroxyproline-rich glycoprotein linked with plant response to pathogen attack and wounding. Tomato plants, cultivar Mountain Fresh, were transformed via Agrobacterium tumefaciens harboring a binary vector for expression of the full-length SN2 gene or ELP cDNA under the regulation of the CaMV 35S promoter. Molecular characterization of PCR-positive putative T-0 transgenic plants by Northern analysis revealed constitutive over-expression of SN2 and ELP mRNA. Junction fragment analysis by Southern blot showed that three of the four SN2 over-expressing T-0 lines had single copies of complete T-DNAs while the other line had two complete T-DNA copies. All four ELP over-expressing T-0 lines had a single copy T-DNA insertion. Semi-quantitative RTPCR analysis of T-1 plants revealed constitutive overexpression of SN2 and ELP. Transgenic lines that accumulated high levels of SN2 or ELP mRNA showed enhanced tolerance to Cmm resulting in a significant delay in the development of wilt symptoms and a reduction in the size of canker lesions compared to non-transformed control plants. Furthermore, in transgenic lines over-expressing SN2 or ELP bacterial populations were significantly lower (100-10,000-fold) than in non-transformed control plants. These results demonstrate that SN2 and ELP over-expression limits Cmm invasiveness suggesting potential in vivo antibacterial activity and possible biotechnological application for these two defense proteins.
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