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Resistance to Alternaria brassicicola in transgenic broccoli expressing a Trichoderma harzianum endochitinase gene

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Abstract

Transgenic broccoli plants expressing a Trichoderma harzianum endochitinase gene were obtained by Agrobacterium tumefaciens-mediated transformation. PCR and Southern blot analysis confirmed the presence of the gene in plants initially selected via resistance to kanamycin. Primary transformants (T0) and selfed progeny (T1) were examined for expression of the endochitinase gene using a fluorometric assay and for their resistance to the fungal pathogens Alternaria brassicicola and Sclerotinia sclerotiorum. All transgenic plants with elevated endochitinase activity had the expected 42 kDa endochitinase band in western blot analysis, whereas no such band was detected in the non-transgenic control. Leaves of most mature T0 plants had 14–37 times higher endochitinase activity than controls; mature T1 plants had higher endochitinase activity (100–200 times that in controls), in part because of lower control values. T0 plantlets in vitro or young plants in soil had higher absolute and relative endochitinase activity. When detached leaves of T0 plants were inoculated with A. brassicicola, lesion size showed a significant negative correlation with endochitinase levels. After inoculation of two-month old T0 plants with A. brassicicola, all 15 transgenic lines tested showed significantly less severe disease symptoms than controls. In contrast, lesion size on petioles of T0 and T1 plants inoculated with S. sclerotiorum was not statistically different from controls.

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  1. A.A. Mora

    Present address: INIFAP-Campo Experimental Bajio, Apdo. Postal 112, Celaya, Gto, 38010, Mexico

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  1. Department of Plant Breeding, Cornell University, Ithaca, NY, 14853-1901, USA

    E.D. Earle

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Mora, A., Earle, E. Resistance to Alternaria brassicicola in transgenic broccoli expressing a Trichoderma harzianum endochitinase gene. Molecular Breeding 8, 1–9 (2001). https://doi.org/10.1023/A:1011913100783

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