Morphogenetic response of cotyledon and leaf explants of melon (Cucumis melo L.) cv. Amarillo Oro

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Abstract

Callus cultures from cotyledon and leaf explants of a Spanish cultivar of melon (Amarillo Oro) were tested for growth and morphogenic capacity on several culture media with different concentrations of IAA (indole-3-acetic acid) in combination with 1.0 mg.L-1 BA (6-benzylaminopurine) or 6.0 mg.L-1 KIN (kinetin). The best results were achieved with cotyledon explants. The leaf explants presented low bud formation capacity. Variability of organogenic response on cotyledons of different age (7, 5, 3 and 1-day-old) was evaluated. The age of explant had a significant influence on bud induction. Cotyledon explants from 7-day-old seedlings showed higher organogenic index and development of shoots when cultured onto MS medium supplemented with 1.5 mg.L-1 of IAA and 1.0 mg.L-1 of BA. The effect of cut type of cotyledonary explants on organogenic response was also investigated. Explants cut transversally showed the best results. The addition of copper sulfate in the culture medium promoted a qualitative improvement of the regenerated shoots.

Translated abstract

Explantes de cotilédones e folhas do cultivar de melão Amarillo Oro foram cultivados para avaliação do potencial morfogenético em diferentes meios de cultura com diversas concentrações de ácido indolacético (AIA), em combinação com 1,0 mg.L-1 benziladenina (BA) e 6,0 mg.L-1 de cinetina (CIN). Os melhores resultados foram obtidos com explantes de cotilédones, sendo que explantes de folhas mostraram uma capacidade baixa na indução de gemas. A variação na resposta organogênica em cotilédones de diferentes idades fisiológicas (1, 3, 5 e 7 dias após a germinação) mostraram que os melhores resultados foram obtidos com cotilédones de sete dias cultivados no meio MS suplementado com 1,5 mg.L-1 de AIA e 1,0 mg.L-1 de BA. O efeito do tipo de corte na resposta organogênica de explantes cotiledonares foi também avaliado. Os melhores resultados foram obtidos com explantes cortados transversalmente. Adição suplementar de sulfato de cobre ao meio de cultura resultou em uma melhora na qualidade da dos brotos regenerados.

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Production of haploid and doubled haploid plants of melon ( Cucumis melo L) for use in breeding for multiple virus resistance.

R. Alan Mitteer, M Lotfi, M J Henning … (2003)

We have developed improved procedures for recovery of haploid and doubled haploid (DH) melon plants, using hybrids derived from crosses of lines with multiple virus resistance. Seeds formed after pollination with irradiated pollen were cultured in liquid medium for 10 days before excision of the embryos for further culture. This made it easier to identify the seeds containing parthenogenetic embryos, thereby reducing the effort required and increasing the percentage of plants recovered. The plants obtained (approximately 175) were transferred to a greenhouse for evaluation. Three fertile lines were identified, and selfed seeds were obtained for evaluating virus resistance. Flow cytometry of leaf tissues showed that two of these lines were spontaneous DH and the third was a mixoploid containing haploid and diploid cells. The other plants remained sterile through the flowering stage. Flow cytometry of 20 sterile plants showed that all were haploid. Attempts to induce chromosome doubling by applying colchicine to greenhouse-grown plants were unsuccessful. Shoot tips from the haploid plants were used to establish new in vitro cultures. In vitro treatment of 167 micropropagated haploid shoots with colchicine produced 10 diploid plants as well as 100 mixoploid plants. Pollen from male flowers that formed in vitro on the colchicine-treated plants was examined. High percentages of viable pollen that stained with acetocarmine were found not only in the diploids but also in >60% of the plants scored as mixoploid or haploid by flow cytometry. Efficient recovery of DH from hybrid melon lines carrying combinations of important horticultural traits will be a valuable tool for melon breeders.