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Tissue Cultures of Maize (Zea mays L.): Initiation, Maintenance, and Organic Growth Factors¹

Callus from 17 of 23 single crosses and 10 of 17 inbreds of Zea mays L. was isolated from mature embryos grown on Linsmaier and Skoog RM medium with 15 mg 2,4-dichlorphenoxyacetic acid (2,4-D) per liter. This callus growth was greatest from the first node of the seedling shoot; occasionally callus growth was observed from the radicle and coleoptile regions. All callus was easily transferred and maintained on media with 2 mg 2,4-D/liter. Genetic variability for callus growth was observed among both single crosses and inbreds. Callus was initiated from 70% of 23 single crosses tested and from only 40% of the 17 inbreds tested. The most rapidly growing cultures were isolated from Oh51A x Os420 which had a 20- to 25-fold fresh-weight increase during a 30-day incubation at 30 C.

Cultures maintained on 2,4-D did not require cytokinins for growth, nor was 2,4-D effectively replaced by idoleacetic acid or -napthaleneacetic acid. Several complex media additives were studied and only 800 mg yeast extract/liter and 200 mg casamino acids/liter individually stimulated significant increases in fresh weight over unsupplemented media. The callus from Oh51A x Os420 has been maintained for 20 months with transfer to fresh media every 30 days (representing 19 subcultures).

Key Words: Hormones • Variability • Inbreds • Single crosses

² Research Associate and Associate Professors, respectively. (Current address for Kleese: Biology Department, Macalester College, St. Paul, MN 55101).

Received for publication June 29, 1973.

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