PNL Volume 17

1985

RESEARCH REPORTS

1 3

IN VITRO CULTURE OF EXPLANTS FROM A SINGLE PEA SEED^1/

Filippone, E.

C.N.R., Centro Studi Miglioramento

Genetico Ortaggi, Portici, Italy

A method was devised to obtain several explants from a single pea

seed as a way of meeting certain needs in breeding, physiological, and

genetical studies.

Seeds of cv 'Proteo' were surface sterilized with calcium hypochlo-

ride (2%, 20 min), rinsed several times with sterile water, and sown on

sterile medium (deionized water plus agar 0.8%) , and then placed in a

growth chamber at 24C in continuous light. Seven-day-old seedlings were

dissected as shown in Fig. 1 to obtain from each: a) root; b) epicotyl

bud; c) epicotyl segment; d) bud with only one cotyledon; e) shoot apex.

All operations were performed under a dissection microscope in a laminar

flow hood. Table 1 lists the media used and the potential uses for the

explants. The explants were incubated in a growth chamber at 24C, under

16 h light/8 h dark. Only the roots were grown in total darkness.

Callus proliferation occurred after 15 days in the epicotyl seg-

ments, and a whole plant was obtained from the bud with only one cotyle-

don. After 40 days each epicotyl bud produced an average of eight

shoots.

Using this technique, a single seed can be used to mass propagate

genetically identical tissue for different studies. Moreover, the coty-

ledon without embryo can be used for analysis ot seed protein.

T. Raggio, M., N. Raggio, and J. G. Torrev. 1957. Amer. J. Botany

44:325-334.

2. Lindsmaier, E. M. and F. Skoog. 1965. Phvsiol. Plant. 18:100-127.

Fig. 1. Explants from a single pea seed.

� Contribution no. 11 from Centro di Studio per il Miglioramento

Genetico degli Ortaggi - C.N.R. - Portici (Napoli) - Italy.