Restoration of fertility in cytoplasmic male-sterile Nicotiana tabacum (cytoplasm N. bigelovii) by protoplast fusion with X-irradiated protoplasts of N. tabacum, SR-1

Raineri, D.; Jordan, P.; Kumar, A.

Journal of Experimental Botany 43(247): 195-203

1992


ISSN/ISBN: 0022-0957
DOI: 10.1093/jxb/43.2.195
Accession: 002210886

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Abstract
The 'donor-recipient' fusion method was used to investigate the intraspecific transfer of organelles and organelle-encoded traits from donor to recipient Nicotiana species under conditions which were selective for chloroplast transfer. An alloplasmic male sterile line of N. tabacum carrying the cytoplasm of N. bigelovii as the recipient and N. tabacum SR-1, a mutant with maternally-inherited streptomycin resistance, as the cytoplasm donor were used. Organelle composition of 13 cybrid plants was investigated by analysis of tentoxin and streptomycin sensitivities, chloroplast and mitochondrial DNA restriction patterns, and alloplasmic male sterility. Chloroplast DNA analysis and the tentoxin test both showed that all 13 cybrid plants possessed chloroplasts from the N. tabacum, SR-1 parent only. Analysis of mitochondrial DNA from second generation plants derived from 11 of the original cybrid plants indicated very little heterogeneity with nine of the plants expressing pure parental-type mtDNAs. Although conditions were selective for donor-type chloroplast transfer, the results indicate the generation of novel cytoplasmic combinations; pure donor chloroplasts in combination with either pure or recombinant-type recipient mitochondria. Our results support previous findings that the 'donor-recipient' fusion method can be used to restore fertility of CMS lines of Nicotiana.