Rearrangement,amplification, and assortment of mitochondrial DNA molecules in cultured cells of Brassica campestris

Summary We compared Brassica campestris mitochondrial and chloroplast DNAs from whole plants and from a 2-year-old cell culture. No differences were observed in the chloroplast DNAs (cpDNAs), whereas the culture mitochondrial DNA (mtDNA) was extensively altered. Hybridization analysis revealed that the alterations are due entirely to rearrangement. At least two inversions and one large duplication are found in the culture mtDNA. The duplication element is shown to have the usual properties of a plant mtDNA high frequency recombination repeat. The culture mtDNA exists as a complex heterogeneous population of rearranged and unrearranged molecules. Some of the culture-associated rearranged molecules are present in low levels in native plant tissue and appear to have sorted out and amplified in the culture. Other mtDNA rearrangements may have occurred de novo. In addition to alterations of the main mitochondrial genome, an 11.3 kb linear mtDNA plasmid present in whole plants is absent from the culture. Contrary to findings in cultured cells of other plants, small circular mtDNA molecules were not detected in the B. campestris cell culture.     

Molecular cloning and characterization of the trpC gene from Penicillium chrysogenum

Summary We cloned the Penicillium chrysogenum trpC gene from a genomic library by complementation of an Escherichia coli trpC mutant lacking phosphoribosylanthranilate isomerase activity. The gene ecodes a 2.7 kb poly(A)⁺ RNA. We localized the gene by sequence analysis in a 2.9 kb DNA insert found in the smallest plasmid selected from the library. Sequence data strongly suggest that the organization of the gene is similar to that described in other Ascomycetes. We found that a DNA fragment which codes only for the carboxy-terminal protion of the polypeptide is sufficient for complementation of the E. coli trpC9830 mutation.     

DNA sequence organization in the genomes of three related millet plant species

Summary A major portion of the genomes of three millet species, namely, barn yard millet, fox tail millet and little millet has been shown to consist of interspersed repeat and single copy DNA sequences. The interspersed repetitive DNA sequences are both short (0.15–1.0 kilo base pairs, 62–64% and long (>1.5 kilo base pairs, 36–38%) in barn yard millet and little millet while in fox tail millet, only long interspersed repeats (>1.5 kilo base pairs) are present. The length of the interspersed single copy DNA sequences varies in the range of 1.6–2.6 kilo base pairs in all the three species. The repetitive duplexes isolated after renaturation of 1.5 kilo base pairs and 20 kilo base pairs long DNA fragments exhibit a high thermal stability with Tms either equal to or greater than the corresponding native DNAs. The S1 nuclease resistant repetitive DNA duplexes also are thermally stable and reveal the presence of only 1–2% sequence divergence.The present data on the modes of sequence arrangement in millets substantiates the proposed trend in plants, namely, plants with 1C nuclear DNA content of less than 5 picograms have diverse patterns of sequence organization while those with 1C nuclear DNA content greater than 5 picograms have predominantly a short period interspersion pattern.Abbreviations kbp kilobase pairs NCL Communication No. 3606.     

Discrimination between closely related Triticeae species using genomic DNA as a probe

Summary Labelled total genomic DNA was used as a probe in combination with blocking DNA to discriminate between taxonomically closely related species in the genera Hordeum and Secale. Discrimination was possible both by Southern hybridization to size-fractionated restriction enzyme digests of genomic DNA and by in situ hybridization to chromosome preparations. To distinguish between two species (e.g. H. vulgare and H. bulbosum), genomic DNA from one species was used as the labelled probe, while unlabelled DNA from the other species was applied at a much higher concentration as a block. The blocking DNA presumably hybridized to sequences in common between the block and the labelled probe, and between the block and DNA sequences on the membrane or chromosomes in situ. If so, mainly species-specific sequences would remain as sites for probe hybridization. These species-specific sequences are dispersed and represent a substantial proportion of the genome (unlike many cloned, species-specific sequences). Consequently, rapid nonradioactive methods detected probe hybridization sites satisfactorily. The method was able to confirm the parentage of hybrid plants. It has potentially wide application in plant breeding for the detection of alien DNA transfer, and it can be easily adapted to many species.     

Tail-to-tail orientation of the Atlantic salmon alpha- and beta-globin genes

We report the cloning of a cDNA and two corresponding -globin genes of the Atlantic salmon (Salmo salar L.) as well as two genes for -globins. Nucleotide sequence analysis of the cDNA shows that the predicted -globin peptide comprises 148 amino acids with a calculated molecular mass of 16,127 Da and an overall amino acid similarity of 40–50% to higher vertebrates and 60–90% to fish sequences. The study of the genomic organization of - and -globin genes shows that, as is the case in Xenopus, the salmon genes are adjacent. Two sets of linked - and -globin genes were isolated and restriction-enzyme polymorphisms indicate that they belong to two distinct loci, possibly as a result of the salmon tetraploidy. In each locus the - and -globin genes are oriented 3 to 3 relative to each other with the RNA coding sequences located on opposite DNA strands. This is the first evidence for this type of arrangement found for globin genes. Moreover, while the linkage found in salmon and Xenopus supports the hypothesis of an initial tandem duplication of a globin ancestor gene, our results raise the question of the actual original orientation of the duplicated genes. Correspondence to: F. Gannon     

Influence of growth rate on pigment and lipid composition of the microalgaIsochrysis aff.galbana clone T.iso

A continuous culture ofIsochrysis aff.galbana clone T.iso, used to feedPecten maximus larvae at IFREMER (Brest, France), was carried out in a chemostat at its optimum temperature for growth (26 °C). Changes in pigments, lipid class (neutral, glyco- and phospholipids) and degree of fatty acid unsaturation were studied at three different growth rates (0.33, 0.5, 1 d^–1). As predicted by chemostat theory, a slow growth rate produced higher cell numbers and higher biomass per unit volume. These cells were low in chlorophylla and carotenoids, but rich in neutral lipids. In contrast, cultures with a fast growth rate yielded lower cell concentrations, buth higher chlorophylla, carotenoid and membrane lipid contents per cell. Changes in polyunsaturated fatty acid distribution were related to differences in algal growth rates. Neutral lipids contained mainly saturated and monounsaturated fatty acids (C18:19) at low growth rates whereas they were enriched in polyunsaturated fatty acids, especially C22:63, at high growth rates. Therefore, it is suggested that the growth rate in continuous cultures be controlled so as to adjust the relative proportions of polyunsaturated fatty acids in lipid classes of the diet meant for larval nutrition.Author for correspondence     

伪狂犬病毒闽A株基因文库的构建及物理图谱分析

本文报道以质粒ｐＢＲ３２２作载体,用鸟枪法克隆出了ＰＲＶ闽Ａ株除ＢａｍＨＩ－１,２外的所有酶切片段,构建了ＰＲＶ闽Ａ株基因文库,并以克隆出的ＢａｍＨＩ片段用光生物素标记作探针,应用分子杂交法确定了ＰＲＶ闽Ａ株绝大部分限制性内切酶位点的位置。     

Rapid changes in amplification and methylation pattern of genomic DNA in cultured carrot root explants (Daucus carota L.)

Summary Rapid genomic DNA variation due to methylation and copy number alteration was observed in carrot root explants 6 h after inoculation and during a 36-h period of exponential callus growth. De novo methylation and amplification of restricted BspNI fragments of low molecular weight occurred before cell cycle activation and should, therefore, be independent of progression through the S-phase of the cell cycle. Growth regulators seemed to influence the amplification pattern indirectly by regulating cell division activity. In exponentially growing callus tissue the copy number of most of the repetitive fragments was dramatically reduced. It is presumed that this reduction in the copy number of repetitive fragments is characteristic of rejuvenilization. 3-Indole-acetic-acid (IAA) and inositol in the medium increased the degree of unspecific genomic DNA methylation in growing rhizogenic carrot callus tissue in the absence of kinetin, which inhibits root induction at that stage. A possible relation to the induction of rhizogenesis is considered. The observed reduction in number of repetitive restriction fragments and the increase in DNA methylation are gross changes covering the total genome. The results are discussed in relation to the controversy concerning the general biological significance of the methylation and amplification of DNA sequences.