Abstract
Viable protoplasts (yield > 1.9 × 107 g−1 fresh weight; mean viability 85±2%, n=5) were isolated from leaves of axenic shoot cultures of Manihot esculenta Crantz. cv. M. Thai 8. Protoplasts were cultured for up to 50 days in liquid, ammonium-free MS medium, overlaying agarose-solidified B5 medium with short glass rods embedded perpendicularly within, and protruding from, the agarose layer. Control protoplasts were cultured identically, but without glass rods. Sustained protoplast division was observed only in the presence of glass rods, where the initial plating efficiency was almost 6-fold greater than control (p < 0.05). The mean final plating efficiency of treated cultures was 1.0±0.2% while, in contrast, significant colony formation was not observed in controls.
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Abbreviations
- BA:
-
6-benzyladenine
- CPPU:
-
N-(2-chloro-4-pyridyl)-N'-phenylurea
- MES:
-
2[N-morpholino]ethane sulphonic acid
- MS:
-
Murashige & Skoog (1962)
- NAA:
-
α-naphthaleneacetic acid
- IPE:
-
initial plating efficiency
- FPE:
-
final plating efficiency
References
Anthony P, Davey MR, Power JB, Washington C & Lowe KC (1994) Synergistic enhancement of protoplast growth by oxygenated perfluorocarbon and Pluronic F-68. Plant Cell Rep. 13: 251–255
Byrne DH (1984) Breeding cassava. In: Janick J. (Ed) Plant Breeding Reviews, Vol 2 (pp 73–134). AVI, Westport
d'Utra Vaz FB, Slamet IH, Khatun A, Cocking EC & Fower JB (1992) Protoplast culture in high molecular oxygen atmospheres. Plant Cell Rep. 11: 416–418
Fellman CD, Read PE & Hosier MA (1987) Effects of Thidiazuron and CPPU on meristem formation and shoot proliferation. HortSci. 22: 1197–1200
Frearson EM, Power JB & Cocking EC (1973) The isolation, culture and regeneration of Petunia leaf protoplasts. Dev. Biol. 33: 130–137
Mathews H, Schopke C, Carcamo R, Chavarriago P, Fauquet C & Beachy RN (1993) Improvement of somatic embryogenesis and plant recovery in cassava. Plant Cell Rep. 12: 328–333
Matsuta N & Hirabayashi H (1989) Embryogenic cell lines from somatic embryos of grape (Vitis vinifera L.). Plant Cell Rep. 7: 684–687
Misawa N, Yamano Y, Ohyama K & Komano T (1982) Protoplast culture of Euphorbia tirucalli and oil body formation in the cells. In: Fujiwara A (Ed) Plant Tissue Culture (pp 607–608). Maruzen, Tokyo
Murashige T & Skoog F (1962) A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 56: 473–497
Shahin EA & Shepard JF (1980) Cassava mesophyll protoplasts: isolation, proliferation and shoot formation. Plant Sci. Lett. 17: 459–465
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Anthony, P., Davey, M.R., Power, J.B. et al. An improved protocol for the culture of cassava leaf protoplasts. Plant Cell Tiss Organ Cult 42, 299–302 (1995). https://doi.org/10.1007/BF00030004
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DOI: https://doi.org/10.1007/BF00030004