Somatic hybridization by fusion of protoplasts 2. the combinations of nicotiana tabacum cultivar bright yellow mutant aurea and nicotiana glutinosa and of nicotiana tabacum cultivar burley 21 and nicotiana alata
Nagao, T.
Japanese Journal of Crop Science 48(3): 385-392
1979
Accession: 006437500
Mesophyll protoplasts of N. tabacum. cv. Bright Yellow mutant Aurea and N. glutinosa and of N. tabacum. cv. Burley 21 and N. alata were prepared from leaf tissue by enzymatic digestion and were fused with the aid of polyethylene glycol. When the fused protoplasts were plated on an agar medium or in a liquid medium for cell culture, new cell walls were regenerated from them and the cell division could be seen beginning on the 14 day of culture in the combination of N. tabacum and N. glutinosa, whereas in that of N. tabacum and N. alata 10th day of culture. Cell division process took place over again after that. After 1 mo. culture, almost all of survived protoplasts grew to cell colonies of 1-2 mm in diameter. Thirty-three somatic hybrid calli of the combination of N. tabacum and B. glutinosa and 2 calli of the combination of N. tabacum and N. alata were obtained. Many plantlets were differentiated from these calli when transferred to enriching zeatin. The chromosome number of the somatic hybrids of the combination of N. tabacum and N. glutinosa was determined: 5 were amphidiploid, 28 showed aneuploid with 50-88 chromosomes. The chromosome number of the somatic hybrids of the combination of N. tabacum and N. alata also was determined: 1 was amphidiploid and the other aneuploid.