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Factors that effect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis

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Abstract

Several factors that affect the frequency of organogenesis in apple leaf explants were examined for the scion cultivars ‘Empire’, ‘Freedom’, ‘Golden Delicious’, ‘Liberty’, ‘McIntosh’, and ‘Mutsu’ and for the rootstocks Malling 7A and Malling 26. The main factors affecting morphogenesis were BA concentration, basal medium, leaf explant origin and maturity, explant orientation, and photosynthetic photon flux. Depending on the genotype, optimal regeneration was obtained using either 22.2 or 31.1 μM BA and the N6 basal medium, with the exception of ‘Golden Delicious’ which regenerated better on MS medium. After 6 weeks, the average number of shoots per segment varied from 5 to 16, and the percentage of regeneration between 70 and 100%, depending on the genotype tested and the maturity of the explant. Regeneration capacity increased dramatically from the tip towards the base of the leaf, and was higher from the middle to the proximal end.

Cefotaxime and carbenicillin, two antibiotics commonly used during transformation studies to eliminate Agrobacterium tumefaciens from plant tissue, were tested to determine their effect on morphogenesis. Cefotaxime at a dose of 250 mg 1-1 enhanced regeneration and shoot development, whereas carbenicillin at a dose of 500 mg l-1 induced abundant callus formation and inhibited regeneration. Kanamycin, a widely used selection agent for plant transformation, strongly inhibited regeneration even at very low doses. Schemes for selection and recovery of transgenic apple plants when kanamycin is used as the selection agent are discussed.

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Abbreviations

BA:

benzyladenine

Cef:

cefotaxime

Crb:

carbenicillin

IBA:

indolebutyric acid

Kan:

kanamycin

LS:

Linsmaier and Skoog (1965) medium

M:

Malling

MS:

Murashige and Skoog (1962) medium

NAA:

naphthaleneacetic acid

N6 medium (Chu et al. 1975):

as modified by Welander (1988)

PPF:

photosynthetic photon flux

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  1. Department of Plant Pathology, Cornell University, NYS Agr. Expt. Station, 14456, Geneva, NY, USA

    Luz Marcela Yepes & Herb S. Aldwinekle

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  1. Luz Marcela Yepes
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  2. Herb S. Aldwinekle
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Yepes, L.M., Aldwinekle, H.S. Factors that effect leaf regeneration efficiency in apple, and effect of antibiotics in morphogenesis. Plant Cell Tiss Organ Cult 37, 257–269 (1994). https://doi.org/10.1007/BF00042339

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