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Effect of Gelling Agent and Selective Sub-culturing on Hyperhydricity in Anther-Derived Coconut Embryos Perera, P. I. P.; Kularatene, J. D. J. S.; Weerakoon, L. K.
International Coconut Community Journal Vol 27 No 2 (2011): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | Full PDF (188.811 KB) | DOI: 10.37833/cord.v27i2.114

Abstract

Experiments were conducted to compare the liquid medium with the media solidified with agar or phytagel. Selective subculturing and use of the embryo maturation medium supplemented with higher concentration of phytagel (0.5%; w/v) were also tested for reducing the vitrified embryos. Modified Eeuwens Y3 medium was used as the basal medium. By culturing the anthers on the medium solidified with phytagel (0.25%; w/v), direct embryo formation (86.7%) and embryo conversion (21.5%) were significantly increased. Plant regeneration efficiency of anther derived embryos or calluses developed in the liquid culture medium was extremely low (2.4%). Vitrification was further reduced by incorporating 0.5% (w/v) phytagel into the embryo maturation medium. Highest plant regeneration efficiency was obtained by exposing the embryos to 0.5% (w/v) phytagel for 21 days, which reduced vitrification by 42%. Furthermore, selective subculturing of the embryos was effective for reducing vitrification.
Viability of heat treated microspores of Cocos nucifera L. for induction of microspore embryogenesis Jayarathna, S. P. N. C.; Suraweera, H. G. M. R.; Piyatissa, N. K. L. S.; Perera, P. I. P.; Vidhanaarachchi, V. R. M.
International Coconut Community Journal Vol 41 (2025): CORD
Publisher : International Coconut Community

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.37833/cord.v41i.465

Abstract

Tall coconut (Cocos nucifera L.) palm is an allogamous and highly heterozygous which is a major obstacle for conventional breeding. Microspore embryogenesis is a promising method which enables the development of homozygous lines. This study is conducted to detect the coconut microspore viability after heat treatment with different incubation periods for the induction of embryogenesis. Anthers were collected from the inflorescences of two coconut palms at three weeks before splitting stage and microspores were extracted. They were subjected to heat treatment for 1, 3, 6 and 9 days either prior or post inoculation. Viability was tested using iodine potassium iodide, acetocarmine and 2, 3, 5- triphenyltetrazolium chloride. Induction of embryogenesis after subjecting to the heat treatment was also studied. Cultures were initiated in modified Eeuwens Y3 medium. To observe the nuclear status of microspores, the samples were stained with 1% acetocarmine after 7 and 14 days. The callus formation was also recorded. A significant difference in viability was observed between control, heat treated prior inoculation and post inoculation when observed in day 01, day 07 and day 14. Significant difference in viability was not detected with no incubation (I0) and 01-day incubation (I1) when microspores were observed in 01 day and 07 days whereas, 3-day incubation (I2), 6-day incubation (I3) and 9-day incubation (I4) resulted significantly different viability of microspores when observed in 01 and 07 day after culturing. Viability of microspores were significantly different for all incubation periods 14 days after culturing. Result was similar with the three stains used to test viability after testing in 1, 7 and 14 days. The highest frequency of binuclear (58.5%) and tetra nuclear (7.2%,) microspores were recorded in the samples treated after culturing the microspores. The multinuclear stages of microspore indicated the induction of embryogenesis. The most effective heat treatment was 38 °C for 3 days after inoculation of the microspores. The results of the study are important for further improvement of microspore embryogenesis protocol.