Section 8
Chapter 7,313

Establishment of embryogenic callus and high protoplast yielding suspension cultures of sugarcane saccharum spp hybrids

Taylor P.W.J.; Ko H L.; Adkins S.W.; Rathus C.; Birch R.G.

Plant Cell Tissue and Organ Culture 28(1): 69-78


ISSN/ISBN: 0167-6857
DOI: 10.1007/bf00039917
Accession: 007312961

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For 18 sugarcane cultivars, four distinct callus types developed on leaf explant tissue cultured on modified MS medium, but only Type 3 (embryogenic) and Type 4 (organogenic) were capable of plant regeneration. Cell suspension cultures were initiated from embryonic callus incubated in a liquid medium. In stage one the callus adapted to the liquid medium. In stage two a heterogeneous cell suspension culture formed in 14 cultivars after five to eight weeks of culture. In stage three a homogeneous cell suspension culture was developed in six cultivars after 10 to 14 weeks by selective subculturing to increase the proportion of actively dividing cells from the heterogeneous cell suspension culture. Plants were regenerated from cell aggregates in heterogeneous cell suspension cultures for up to 148 days of culture but plants could not be regenerated from homogeneous cell suspension cultures. High yields of protoplasts were obtained from homogeneous cell suspension cultures (3.4 to 5.2 .times. 106 protoplasts per gram fresh weight of cells [gfwt-1]) compared to heterogeneous cell suspension cultures (0.1 .times. 106 protoplasts gfwt-1). Higher yields of protoplasts were obtained from homogeneous cell suspension cultures for cultivars Q63 and Q96 after regenerating callus from the cell suspension cultures, then recycling this callus to liquid medium (S-cell suspension cultures). This process increased protoplast yield to 9.4 .times. 106 protoplasts gfwt-1. Protoplasts isolated from S-cell suspension cultures were regenerated to callus and recycled to produce SP-cell suspension cultures yielding 6.4 to 13.2 .times. 106 protoplasts gfwt-1. This recycling of callus to produce S-cell suspension cultures allowed protoplasts to be isolated for the first time from cell lines of cultivars Q110 and Q138.

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