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Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass



Somatic embryogenesis and plant regeneration from embryogenic suspension cultures of perennial ryegrass



In Vitro Cellular & Developmental Biology 26(4): 419-424



Embryogenic callus induced from mature caryopses of perennial ryegrass (Lolium perenne L.) were placed in liquid half-strength Murashige and Skoog (MS) basal medium and supplemented with 6.0 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D), 3 g/l (w/v) casein hydrolysate (CH), and B5 vitamins, to initiate fast-growing highly embryogenic cell suspension cultures. Newly initiated suspension cultures contained a high level of large non-embryogenic cells (NE) with relatively few embryogenic (E) cells. Cell types were separated by discontinuous Percoll gradients or by filtering the newly initiated cultures through 31-.mu.m nylon mesh. The growth conditions of the E cells were optimized by testing various media components including 2,4-D and sucrose, and subculture dilution ratio. Optimal shoot formation occurred after pretreatment of the embryogenic cells on solidified callus maintenance medium supplemented with 60 mg/l cefotaxime for 4 weeks prior to transfer to regeneration medium. Regeneration media consisted of half-strength MS basal medium supplemented with B5 vitamins, 0.5 mg/l fluridone, and 0.5 mg/l BA. Most plants regenerated were albino with only a few green plants.

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Accession: 002224241

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DOI: 10.1007/bf02623834


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