PAG-I Plant Genome I Conference

Town & Country Conference Center, San Diego, CA, November, 1992.


PG-I: 69pg1

COMPLEX GENE FAMILIES OF LOBLOLLY PINE.

Claire S. Kinlaw and Suzanne Gerttula. Institute of Forest Genetics, Pacific Southwest Research Station, USDA Forest Service, PO Box 245, Berkeley, California, 94701.


Conifer genomes in general, and pine genomes in particular, are among the largest in the plant kingdom. For example, loblolly pine (Pinus taeda) contains 10pg DNA per haploid genome as compared to 0.1 pg per Arabidopsis haploid genome. Conifer genomes, like angiosperm genomes, contain much repeated sequence DNA. Unlike angiosperms, a significant part of the repeated pine DNA may fall into complex gene families. As a result of the pine genome mapping project (David Neale and co-workers), it has been observed that many random cDNAs hybridize to multiple bands on Southern blots. "Single copy" banding patterns are only rarely observed for loblolly pine cDNAs, and approximately 25% of all random cDNAs from loblolly pine seedling mRNA hybridize to more bands than can be easily counted (>l0). We have begun an investigation into the primary structure, function, genome organization, and evolution of a number of these complex pine gene families. We have performed Southern blots with copy number reconstructions to estimate a minimum gene family sizes, and we have compared Southern banding patterns between loblolly pine a hard pine (Pinus subgenus of the genus Pinus) and western white pine (Pinus monticola) a soft pine (Stobus subgenus of the genus Pinus). Gene families sizes for the clones chosen, range from 4 to approximately 20. Hybridization intensities vary among Southern bands for each particular cDNA, suggesting considerable sequence heterogeneity among gene family members. Hybridization of loblolly pine cDNAs to western white pine genomic DNA vs. loblolly pine genomic DNA results in fewer bands of lower intensity, suggesting that some members of pine complex gene families have diverged beyond detectable limits between the two halves of the pine genus. We have determined the mRNA sizes from northern blots and DNA sequence for each cDNA, and used the DNA sequence to compare to DNA sequences from GenBank. Approximately 10% of the cDNAs analyzed show clear sequence similarity to plant sequences in GenBank. One particular clone, IFGp2027, appears to be a non-specific pine lipid transfer protein, based upon nucleotide sequence similarity, conservation of key amino acids, small size, basic pi, and similarity of hydropathy plots. This protein has been shown to be under developmental, cell specific, and environmental control in angiosperms (S.Torres-Schumann, Plant Mol. Biol. 18: 749-757). In loblolly pine, this lipid transfer protein is a highly abundant message in seedlings, perhaps as high as 10% of the poly A+ RNA.


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