CBN-V Video Archives - S7-21
In vitro Culture of Isolated Pollen of Cassava: Influence of Boric Acid and Growth Regulators in Pollen Embryogenesis

Mukherjee A. and S.K. Naskar

Regional Centre of CTCRI, Dumduma Housing Board, Bhubaneswar-19, Orissa, India archanarcctcri@yahoo.com

        Culture of isolated pollens is advantageous compared to culturing the whole anther for haploid induction. Studies on isolation of pollens from flower buds, pollen viability and response of pollens in in vitro cultures were carried out in four genotypes of cassava. Pollen was isolated from male flower buds at different stages of maturity through maceration, centrifugation and filtration processes. Of the various concentrations of sucrose and maltose used for maceration, maximum density 7000-12000 microspores/ml of pollen mass was obtained with combination of 3% sucrose and 1% maltose. Under similar conditions, pollen viability was recorded as 52-76% among the four genotypes. Pollen cultures raised in liquid MS medium supplemented with 2,4-D (0.5 – 1 mg/l) produced callus on subculture to solid media containing BAP (1- 2 mg/l), NAA (0.5 – 1 mg/l), GA3 (0.2 – 0.5 mg/l) and boric acid (50-100 mg/l). Histo-morphological screening of callus cultures showed development of proembryoids. Though callusing was more in the media containing NAA, frequency of proembryoids was quite low (<0.5%). Despite differential responses to pollen embryogenesis among the genotypes, use of boric acid in combination with other growth regulators was found quite effective (1-1.5%). These results would help in monitoring further growth and morphogenesis of the proembryoids developed from pollen for induction of haploidy.

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