Plant development in long-term embryogenic callus lines of Cucurbita pepo

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Abstract
Embryogenic callus derived from pumpkin hypocotyl segments was induced and maintained for 15 years on MS medium supplemented with the auxins IBA (4.9 μM), 2, 4-D (4.5 μM) or IAA (5.7 μM). On induction media continued embryo maturation and development of adult plants typically failed. Therefore, small embryogenic clumps and individually isolated embryos were subcultured two to four times on one of the conversion media: MS supplemented with 1.5% sucrose and (a) no hormone, (b) 2.9 μM IAA, (c) 5.7 μM IAA, (d) 11.4 μM IAA, (e) 12 μM IEt, (f) 3.8 μM ABA or (g) 2% activated charcoal. The cell line and the kind of auxin used in the induction and maintenance medium, both had a marked influence on the development of plantlets. The best result was achieved with a line that has been induced and maintained for 15 years on MS with IBA. In the IBA line, out of 100 embryos, 77 developed into plantlets on MS medium supplemented with 11.4 μM IAA.