Figure 6. PCR analysis of an F2 tomato population segregating for the fa phenotype.
DNA from wild-type +/+ (1), heterozygous +/fa (2) and mutant fa/fa (3) plants was subjected to PCR with 33P-labelled TOFL specific primers, and resolved in a 5% polyacrylamide gel. The deleted TOFL allele detected in fa plants co-segregates with the fa phenotype in a total of 240 plants analysed.
To establish the spatial and temporal expression pattern of TOFL, we performed in situ hybridization at various stages of wild-type plant development ( Fig. 7). During the initial vegetative phase, TOFL transcripts were found in the apical meristem, around provascular bundles of the stem, in leaf primodia and leaves, and in axillary buds ( Fig. 7a). Expression in leaves was higher at the adaxial face and towards the margins. Serial section of the vegetative meristem indicated that TOFL transcripts accumulate in the three cell layers that involve the meristem but not in the internal zone ( Fig. 7b). This expression pattern agrees with that previously described by Pnueli et al. (1998) for a putative LFY homologue in tomato, although they did not show the gene sequence. In order to determine the temporal expression of TOFLduring the vegetative phase of development, the vegetative meristem of plants that had developed either two, four or six leaves was dissected and used for in situ hybridization. The gene was early detected in the vegetative meristem of plants with just two leaves, as well as in leaves and axillary meristems.
Figure 6. PCR analysis of an F2 tomato population segregating for the fa phenotype.
DNA from wild-type +/+ (1), heterozygous +/fa (2) and mutant fa/fa (3) plants was subjected to PCR with 33P-labelled TOFL specific primers, and resolved in a 5% polyacrylamide gel. The deleted TOFL allele detected in fa plants co-segregates with the fa phenotype in a total of 240 plants analysed.
To establish the spatial and temporal expression pattern of TOFL, we performed in situ hybridization at various stages of wild-type plant development ( Fig. 7). During the initial vegetative phase, TOFL transcripts were found in the apical meristem, around provascular bundles of the stem, in leaf primodia and leaves, and in axillary buds ( Fig. 7a). Expression in leaves was higher at the adaxial face and towards the margins. Serial section of the vegetative meristem indicated that TOFL transcripts accumulate in the three cell layers that involve the meristem but not in the internal zone ( Fig. 7b). This expression pattern agrees with that previously described by Pnueli et al. (1998) for a putative LFY homologue in tomato, although they did not show the gene sequence. In order to determine the temporal expression of TOFLduring the vegetative phase of development, the vegetative meristem of plants that had developed either two, four or six leaves was dissected and used for in situ hybridization. The gene was early detected in the vegetative meristem of plants with just two leaves, as well as in leaves and axillary meristems.
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