Evaluation of Transgenic Anthuriums Expressing the Shiva-1 Fene Encoding a Synthetic Antimicrobial Peptide

Date
2002
Authors
Fujii, Todd M
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Abstract
The effect of anthuriums genetically engineered to express a secreted cercropin analogue Shiva-1 on bacterial blight, caused by Xanthomonas campestris pv. diejfenbachiae and non-target bacteria species was determined. Two cultivars of blight susceptible anthuriums, ‘Paradise Pink’ (formerly labeled ‘Marian Seefiirth’) and ‘Tropic Flame’, actively expressing the Shiva-1 transgene were disease-challenged with a bioluminescent strain of Xanthomonas campestris pv. diejfenbachiae {Xcd Lux). Bioluminescence produced by infected leaves was captured on X-ray film enabling the monitoring and quantification of infection. Disease severity as percentage leaf area infected with Xcd Lux was compared among replicated transgenic lines and nontransgenic control plants over three months. Two transgenic lines of ‘Paradise Pink’, MS 1-2 and MS 1-4, displayed enhanced resistance to the anthurium bacterial blight. At the termination of the experiment disease severity of MS 1-2 and MS 1-4 was significantly less (Pr > F = 0.0001) than the naturally resistant cultivar ‘Kalapana’. One transgenic line of ‘Tropic Flame’, UH 712 1-16, exhibited enhanced susceptibility to anthurium bacterial blight. Disease severity of UH 712 1-16 was significantly higher (Pr > F = 0.0038) than the control line and other transformed lines of the same cultivar. No enhanced resistance was observed among other transgenic UH 712 lines and the control. In order to understand if biological control strategies are to be used in conjunction with genetic resistance, the effect of transgenic anthuriums producing the Shiva-1 lytic peptide on beneficial bacteria was also studied. Inhibitory concentrations of Shiva-1 against four foliar biocontrol bacteria (BCAs) and Xcd were determined by dilution plating after 18 hours exposure to the lytic peptide. Populations were measured in guttation fluid 6 and 11 days post-inoculation. Two of four BCAs, Gut 3 and Gut 6 were statistically less sensitive to Shiva-1 then Xcrf (P < 0.05); Xcd was completely inhibited at 0.5 micromolar. Populations of Gut 3 and Gut 6 increased at 1 micromolar Shiva-1. Transgenic anthuriums, ‘Paradise Pink’, MS 1-1, MS 1-5 and ‘Mauna Kea’, MK 1-2, MK 2-6 did not inhibit BCAs more than non-transgenic anthuriums. Biological control with the BCAs, Sphingomonas chlorophenolica, Microbacterium testaceum, Brevundimonas vesicularis, and Herbaspirillium rubrisubalbicans is thus compatible with engineered genetic resistance to bacterial blight on anthurium.
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