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Southern blots of mitochondrial (mt) DNAs of three Lupinus species cleaved with three restriction enzymes were probed with Lupinus luteus mtDNA fragments containing 18S, 5S rRNA genes or a tRNA-like repeat. Comparison of the number of hybridizing bands and their intensity suggested that the mt 18S and 5S rRNA genes occur mostly in one copy in the genomes of three lupin species. The exception concerned the Lupinus angustifolius 5S rRNA gene showing two hybridizing bands of unequal intensity. The results of hybridization of the lupin mitochondrial genomes with a probe specific for the Lupinus luteus tRNA-like repeat pointed to the presence of such a repeat in other parts of the genomes besides the vicinity of the 18S rRNA gene. Northern hybridization analysis showed the presence of 18S, 5S and tRNA-like repeat transcripts similar in size in all lupin species.
All higher plant plastid genomes have six classes of tRNA genes containing introns. One of those is the tRNALys UUU gene, which encodes maturase protein. In the case of liverwort species from the genus Porella and mosses from the genus Plagiomnium, the maturase coding gene (matK) represents a truncated form of other plant matK genes: several subdomains of the reverse transcriptase-like domain and so-called domain X are not present in these ORFs. These ORFs probably represent pseudogenes of the matK gene. The analysis of codon usage within the matK gene revealed the presence of strong A/T pressure. The use of codons with the third letter being U or A varies from 71-93%. The comparison of maturase amino acid sequences at the family level shows a high identity between species. However, when liverwort and angiosperm maturase sequences are compared, the percentage of identity drops dramatically. The calculated values of the number of nucleotide substitutions vary considerably, even when liverwort species are compared pairwise. The phenetic tree of relationships between plant species on the basis of tRNALys UUU intron sequences concur with the generally accepted plant phylogeny.
A nuclear DNA fragment (7.8 kb) from yellow lupin (L. luteus) was sequenced and shown to contain tRNA(Gly) (GGC) genes and tRNAGly (GGC) pseudogenes organized in three tandemly repeated units: of 2565 bp and 2564 bp, and one, truncated from its 3' end, of 1212 bp. Each unit contains an identical pair of a tRNA(Gly) gene and a pseudogene, both having the same polarity. The nucleotide sequence of the gene appears colinear to L. luteus cytoplasmic tRNA(Gly) (GGC) primary structure. All three genes are efficiently transcribed in HeLa-cell nuclear extract giving two primary transcripts. The main, longer primary transcripts have each an extremely long 3' trailer of about 100 nucleotides, the structure of which is specific only for tRNAGly genes and pseudogenes (80% homology) of the studied tandem (but not for other tRNA(Gly) genes of the yellow lupin genome) as it has been shown by Southern hybridization. This distinctive feature allowed to isolate putative tRNAGly precursor(s) encoded by at least one of the three tRNA(Gly) (GGC) genes from L. luteus seedlings.
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