Production of asymmetric somatic hybrid plants between Cichorium intybus L. and Helianthus annuus L. |
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Authors: | S Varotto E Nenz M Lucchin P Parrini |
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Institution: | (1) Department of Environmental Agronomy and Crop production, University of Padova, Agripolis Via Romea 16, 35020 Legnaro (PD), Italy e-mail: svarotto@agripolis.unipd.it, IT |
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Abstract: | In order to obtain male-sterile asymmetric somatic hybrids between chicory (Cichorium intybus L.) and a sunflower (Helianthus annuus L.) male-sterile cytoplasmic line, mesophyll chicory protoplasts inactivated with iodoacetic acid and hypocotyl sunflower
protoplasts irradiated with γ-rays have been fused, using PEG and applying two different procedures. Thirty three plants were
regenerated from putative hybrid calli. A cytological analysis of their root-tip cells indicated that most of them had 18
chromosomes, the same number as chicory. Through Southern hybridisation on total DNA using the maize mitochondrial specific
gene probes Cox I, Cox II and Cob, three plants were identified as cytoplasmic asymmetric hybrids, as shown by hybridisation
bands specific for both chicory and sunflower. One of the regenerated plants produced a novel pattern of hybridisation that
was not detected in either parent. When hybridisation of total DNA was carried out with an atpA mitochondrial gene probe the
same three cybrids presented both the fertile chicory fragment and the male-sterile sunflower fragment. Finally, Southern
hybridisation with an ORF 522 probe, which in sunflower is co-transcribed with the atpA gene, confirmed the hybrid nature
of the three plants. The morphology of the cybrids resembled the parental chicory phenotype, and at anthesis their anthers
produced fewer pollen grains which could not germinate either ”in vitro” or ”in situ.” Cybrid plants grown in the field produced
seeds when free-pollination occurred.
Received: 26 April 2000 / Accepted: 28 August 2000 |
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Keywords: | Cichorium intybus L Helianthus annuus L Asymmetric somatic hybrid Cytoplasmic male sterility Protoplast fusion |
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