Ecology, Environment and Conservation Paper

Vol 24, Issue 1 2018; Page No.(145-152)

SOMATIC HYBRIDIZATION OF CELLS BETWEEN CITRUS AND MURRAYA PANICULATA BY ELECTRO-FUSION

Hasan Basri Jumin

Abstract

Protoplasts isolated from embryogenic callus of Citrus sinensis were electrically fused with embryogenic protoplasts isolated from seedless Citrus relatives. Hybrid of somatic embryos were obtained after 2 months of culture. The somatic hybrid embryos were obtained by screening on the basis of chromosomes count. The number of chromosome of embryogenic callus counting revealed plantlets tetraploids (2n = 4x = 36) and the other were diploids (2n = 2x = 18). Percentage of multinucleat of protoplast increasing linearly with increase the AC voltage / cm from 15 volt /cm until 45 volt/cm, and decreasing significantly after 60 volt/cm. Number of broken protoplast significant increased linearly until 250 DC volt / cm, and decreased afterward, however the total fusion was only increased until 200 DC volt/cm. The effect of AC field strength on percentage of overall fusion and percentage of heteronucleat was shown a positive effect where the percentage of fused protoplast increased with the same trend with percentage of heteronucleat. The high number of total fusion of protoplast was reached on 30 volt /cm and decreased after 45 volt /cm. This somatic hybrid after regenerated into plants will be utilized as a possible pollen parent for improving the Citrus sinensis.

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