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Callus Induction, Maintenance and Plantlet Regeneration in Creeping Bentgrass¹

Callus induction, from mature caryopses, and callus maintenance and plantlet regeneration, using 14-month-old stock callus (SC), were investigated in Penncross creeping bentgrass (Agrostis palustris Huds.). Greatest callus induction occurred on Murashige and Skoog (MS) medium with 1.0 mg 2,4-dichlorophenoxyacetic acid (2,4-D) under light incubation and 1.0 or 10.0 mg 2,4-D/1 under dark incubation. Kinetin supplements were not required for callus induction, however, caryopses incubated in the dark with 0.01 mg kinetin had greater callus formation than caryopses incubated without kinetin when combined with 1.0 mg 2,4-D/1. Under dark incubation, caryopses produced the greatest callus yields (47 and 57 mg) and callus removal from caryopses required less time and effort. Callus induced from caryopses originated from the scutellum, root cortex and along the coleoptile up to and adjacent the first node. Callus growth using SC was greatest at 1.0 mg 2,4-D/1. Callus previously cultured on 1.0 mg 2,4-D maintained good plantlet regeneration capacity; whereas, callus previously cultured at 10 and 100.0 mg 2,4-D/1 had poor and no plantlet formation, respectively. The best combination for shoot and root development in SC occurred at 0.10 mg and 1.0 mg kinetin/1 without 2,4-D. All plantlets were potted in a sand:peat (1:1) mix and maintained in a glasshouse. Callus induced plants were similar to seeded plants. Less than 1% of the plantlets formed were albino.

Key Words: Tissue culture • Agrostis palustris • Huds • Organogensis • 2,4-dichlorophenoxyacetic acid • Kinetin • Turfgrass

² Associate professor of agronomy, graduate assistant and former graduate assistant, Mississippi State Univ. Mississippi State, MS 39762.

Received for publication September 25, 1981.