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The Maize lpa241 Mutation Causes a Remarkable Variability of Expression and Some Pleiotropic Effects

^a Dipartimento di Produzione Vegetale, Università degli Studi di Milano, Via Celoria 2, 20133 Milano, Italy
^b Dipartimento di Scienze Biomolecolari e Biotecnologie, Università degli Studi di Milano, Via Celoria 26, 20133 Milano, Italy
^c Dipartimento di Genetica e Microbiologia, Università degli Studi di Pavia, Via Ferrata 1, 27100 Pavia, Italy

View larger version (8K): [in a new window]   Fig. 1. Schematic representation of the structure of the 3618-bp lpa241 MIPS1S coding sequence that was cloned in a Blunt II-TOPO vector and sequenced. Black boxes indicate the exons. Comparison of the lpa241 MIPS1S gene with the wt MIPS1S gene (accession no. AF323175) shows 10 single nucleotide mismatches.

View larger version (96K): [in a new window]   Fig. 2. Seed free phosphate levels in the lpa241 families. Individual mature lpa241 homozygous seeds from each of 20 Lpa241/lpa241 self-pollinated families (BC2F2) were analyzed for inorganic phosphate. The free P is expressed as P concentrations. Results are means of four independent assays ± standard deviation (SD). Statistical analysis reveals significant differences among families (ANOVA with; df = 79; F = 18.64; P < 0.01).

View larger version (84K): [in a new window]   Fig. 3. Cleaved amplified polymorphic sequence marker for the MIPS1S gene. A 458-bp MIPS1S amplification product, obtained using the Zm1302 + Zm1580 primers, showed MscI restriction differences that were able to distinguish –/– lpa241 MIPS1S ACR gene (fragments, 292 bp + 166 bp) from +/+ Lpa241 MIPS1S B73 allele (fragments, 133 bp + 159 bp + 166 bp).

View larger version (94K): [in a new window]   Fig. 4. Thin layer chromatography analysis of the "strong" lpa241 expression mutant. Chromatograms were obtained by running trichloroacetic acid extracts from mature single seeds. Inositol esakis phosphate (Ins P6) 20 nmol and inorganic Phosphate (Pi) 10 nmol, in aqueous solution, were loaded as standards.

View larger version (86K): [in a new window]   Fig. 5. (A) Effect of lpa241on seed development (appearance of embryo specific mutants, emb). (B) ears development and (C) plant growth 25 d after sowing. The reference wild-type line is B73.

View larger version (175K): [in a new window]   Fig. 6. Longitudinal section of mature lpa241 (B) and wild-type (A) kernels stained with saphranine-fast green (ed, endosperm; sc, scutellum; sh, shoot; rt, root). Bar: 500 µm.

View larger version (215K): [in a new window]   Fig. 7. Longitudinal sections of mature lpa241 (B, D and F) and wild-type (A, C and E) kernels stained with toluidine blue. Some of the globoids are marked with arrowheads (ed, endosperm; sc, scutellum). Bar: 50 µm.