DEVELOPMENT OF HYBRIDS OF PLEUROTUS FLORIDA AND VOLVARIELLA VOLVACEA THROUGH PROTOPLAST FUSION
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Date
2013
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Abstract
Protoplast fusion using Polyethylene glycol (PEG) and electrofusion was carried out between strains
of Pleurotus florida and Volvariella volvacea to develop improved hybrids. Preliminary screening was
carried out based on responses to growth at different temperature regimes, colony radial growth,
morphology and growth patterns of colony, hyphal size, presence or absence of clamp connections,
and other unique characters like production of reddish exudates. Following the rigorous screening,
eight intergeneric hybrids (Pv14, Pv19, Pv21, Pv27, Pv45, Pv66, Pv102 and Pv127) were selected and
further evaluated for fruit body formation. Pv45, Pv66 and Pv102 were unable to give primordia after
25 days of growth and were considered sterile. Among those which developed fruit bodies, Pv21 and
Pv127 had brownish circular caps with regular margins and central stipes. Other strains showed cap
morphologies closely related to PF5.
Isozyme (alcohol dehydrogenase, malate dehydrogenase, esterase and super oxide dismutase) pattern
analyses indicated presence of bands in all the strains which verified hybridization between the two
parents. Besides, two bands observed neither in the parents nor in the other strains were noticed in
pv27 and pv127. Amplifications of genomic DNA from all the strains using 14 arbitrary decamer
RAPD primers resulted in 98 bands, with size range of 160bps to 2500bps, of which 57.14 per cent
were polymorphic. The bands were obtained only from 8 primers. A combined clustering analysis of
these bands using NTSYSpc software indicated that Pv127 exhibited marked divergence from the
other strains in the phylogenetic clusters and had better performance. Therefore, the salient
investigation of the current work was screening of a hybrid strain (Pv127) with high BE (52.71%) and
low temperature tolerance compared to V. volvacea
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