Callus Induction, Maintenance and Plantlet Regeneration in Creeping Bentgrass¹

Abstract

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.

Long-term callus cultures of diploid Barley (Hordeum vulgare). I. Auxin effects on culture initiation and maintenance

“…While 2,4-D is necessary for effective callus initiation and production, cytological and genetical abnormalities are known to result (Sunderland, 1977 ;Deambrogio & Dale, 1980) . Along with these, high concentrations of 2,4-D have also been found to inhibit subsequent shoot formation (Krans et al ., 1982 ;Torello & Symington, 1984) .…”

Somatic embryogenesis and plant regeneration from cultured mature caryopses of bahiagrass (Paspalum notatum Flugge)

“…Plant regeneration of temperate [1,2,3,4] and tropical forage grasses [5,6] via somatic embryogenesis has been reported. Little work has been reported on bahiagrass (Paspalum notatum Flugge).…”

“…Vasil & Vasil [7] suggested that young meristematic inflorescences of Gramineae were most useful for in vitro culture. While the choice for Gramineae explants has been young inflorescences [1,4,8,9,11], leaves [3,12], immature embryos [13] and mature caryopses [2,4,14,15] have also been used to produce somatic embryos. Since bahiagrass flowering is restricted to June-September it would be advantageous to use plant parts other than immature inflorescences to initiate callus cultures.…”

Tissue culture and long-term regeneration of Phragmites australis (Cav.) Trin. ex Steud.

“…Also, the linkage of culture morphology to regeneration has shown a distinct embryogenic callus phenotype in the Gramineae [7,8]. We chose to work with juvenile tissue from germinated seeds, given the successful development of high-frequency regenerable tissue cultures from immature leaves [9], immature embryos [10], immature inflorescence [8,11] and mature seeds [12] of graminoids.…”