Transmission Of Mitochondrial DNA Research Articles

Populations of plastids and mitochondria during male reproductive cell maturation in Nicotiana tabacum L.: A cytological basis for occasional biparental inheritance

The dynamics of plastid and mitochondrial populations in male reproductive cells of tobacco (Nicotiana tabacum L.) were examined during development using serial ultrathin sections and transmission electron microscopy to reconstruct 58 generative cells and 31 sperm cells at selected stages of maturation from generative cell formation through gametic fusion. The first haploid mitosis resulted in incomplete exclusion of plastids providing an average of 2.81 plastids and 82.7 mitochondria for each newly formed generative cell. During generative-cell maturation, plastid content decreased to an average of 0.48 plastids/generative cell at anthesis owing to autophagy of organelles. Plastids were present in low frequency within generative and sperm cells in the pollen tube and appeared to be transmitted, according to observations immediately prior to fertilization. This forms a cytological basis for genetic reports of occasional biparental plastid inheritance. In contrast, mitochondria were transmitted in larger numbers, and approximately 80 mitochondria per generative cell or sperm cell pair were retained throughout development. This provides a potentially stable source for the transmission of male mitochondrial DNA, if present at fertilization.

Maternal inheritance of chloroplast genome and paternal inheritance of mitochondrial genome in bananas (Musa acuminata)

Restriction fragment length polymorphisms (RFLPs) were used as markers to determine the transmission of cytoplasmic DNA in diploid banana crosses. Progenies from two controlled crosses were studied with heterologous cytoplasmic probes. This analysis provided evidence for a strong bias towards maternal transmission of chloroplast DNA and paternal transmission of mitochondrial DNA in Musa acuminata. These results suggest the existence of two separate mechanisms of organelle transmission and selection, but no model to explain this can be proposed at the present time. Knowledge of the organelle mode of inheritance constitutes an important point for phylogeny analyses in bananas and may offer a powerful tool to confirm hybrid origins.

Localization of a cruciform cutting endonuclease to yeast mitochondria.

We have found a cruciform cutting endonuclease in the yeast, Saccharomyces cerevisiae, which localizes to the mitochondria. This activity apparently is associated with the mitochondrial inner membrane since the activity is not released into solution by osmolysis, in contrast to the matrix enzyme, isocitrate dehydrogenase. The cruciform cutting activity appears to be encoded by CCE1. This gene has been shown to encode one of the major cruciform cutting endonucleases present in yeast cell. In cce1 strains, which lack CCE1 endonuclease activity, the mitochondrial cruciform cutting endonucleolytic activity is also absent. Since CCE1 is allelic to MGT1, a gene required for the highly biased transmission of petite mitochondrial DNA in crosses between rho+ and hypersuppressive rho- cells, it seems likely that the CCE1 endonuclease functions within mitochondria.

Male transmission of linear plasmids and mitochondrial DNA in the fungus Neurospora.

One of the general rules of heredity is that in anisogamous matings genetic elements in organelles are inherited maternally. Nevertheless, there are cases of paternal transmission, both as rare exceptions, and as regular modes of inheritance. We report two new cases of paternal transmission in crosses of the model fungus Neurospora. First, we show leakage of a linear plasmid from males, the first case in fungi and the second in eukaryotes. Transmission frequencies ranged from 1% to 15% in different crosses, but some crosses showed no detectable male transmission. Second, we show leakage of male mitochondrial DNA (mtDNA), the second case in fungi. Some of the resulting progeny have only the male mtDNA type, but some are heteroplasmons. Heteroplasmons show novel restriction fragments attributable to recombination or rearrangement. Heteroplasmy of mtDNA through male transmission has not been reported previously in any eukaryote. In addition we have shown paternal leakage of circular mitochondrial plasmids, supporting another reported case. In a male bearing a linear and a circular plasmid, these plasmids and the mtDNA are transmitted in different combinations. These results show a potential for mitochondrial segregation and assortment during the sexual cycle in anisogamous fungi, pointing to more potential avenues for novel associations between genomic compartments, and between genomic and extragenomic elements.

Uniparental inheritance and replacement of mitochondrial DNA in Neurospora tetrasperma.

This study tested mechanisms proposed for maternal uniparental mitochondrial inheritance in Neurospora: (1) exclusion of conidial mitochondria by the specialized female reproductive structure, trichogyne, due to mating locus heterokaryon incompatibility and (2) mitochondrial input bias favoring the larger trichogyne over the smaller conidium. These mechanisms were tested by determining the modes of mitochondrial DNA (mtDNA) inheritance and transmission in the absence of mating locus heterokaryon incompatibility following crosses of uninucleate strains of Neurospora tetrasperma with trichogyne (trichogyne inoculated by conidia) and without trichogyne (hyphal fusion). Maternal uniparental mitochondrial inheritance was observed in 136 single ascospore progeny following both mating with and without trichogyne using mtDNA restriction fragment length polymorphisms to distinguish parental types. This suggests that maternal mitochondrial inheritance following hyphal fusions is due to some mechanism other than those that implicate the trichogyne. Following hyphal fusion, mutually exclusive nuclear migration permitted investigation of reciprocal interactions. Regardless of which strain accepted nuclei following seven replicate hyphal fusion matings, acceptor mtDNA was the only type detected in 34 hyphal plug and tip samples taken from the contact and acceptor zones. No intracellular mtDNA mixtures were detected. Surprisingly, 3 days following hyphal fusion, acceptor mtDNA replaced donor mtDNA throughout the entire colony. To our knowledge, this is the first report of complete mitochondrial replacement during mating in a filamentous fungus.

Temperature-dependent selection in the transmission of mitochondrial DNA in Drosophila

Temperature-dependent selection in the transmission of mitochondrial DNA in Drosophila

Temperature-dependent selection in the transmission of mitochondrial DNA in Drosophila.

We previously reported a selective mode of mitochondrial DNA (mtDNA) transmission in mtDNA heteroplasmy that was induced artificially in Drosophila melanogaster; the transmission bias appeared to depend on the particular temperature at which heteroplasmic lines were maintained. Here we report investigations of the temperature-dependent mode of mtDNA transmission in heteroplasmic lines for intra- and interspecific combinations maintained separately at 22.5 degrees C, 25 degrees C and 29 degrees C for 20 generations. We have examined a selection model for mitochondrial transmission, similar to genetic selection in haploid organisms. Changes in the relative proportions of two types of mtDNA fit the expectations from the model well. The intensity of selection estimated as a selection coefficient depends on temperature. Temperature-sensitive processes thus appear to be involved in the transmission and maintenance of mitochondria.

Transmission, recombination and conversion of mitochondrial markers in relation to the mobility of a group I intron in Chlamydomonas.

Mitochondrial DNA transmission has been analyzed in diploids produced from sexual crosses or artificial fusions between Chlamydomonas strains which differ by several genetic markers: a group I intron (Cs cob. 1 or alpha intron), three restriction sites (Nh, Nc and H markers) located 0.5-5 kb from the insertion site of the intron, and a MUD2 point mutation (27 bp from the insertion site) conferring resistance to myxothiazol. Recombination between mitochondrial markers is a general property of all crosses and fusions analyzed. In crosses between two intron-containing (alpha+) strains or two intron-less (alpha-) strains, the transmission is preferentially paternal (mt-), with a preponderance depending on the nature of the parental genomes. In crosses between alpha+ and alpha- strains, the conversion of intron-less molecules intron+ is frequent when the alpha+ parent is maternal (mt+) and nearly absolute when the alpha+ parent is paternal (mt-). In 94% of cases, the conversion is accompanied by the co-conversion of the MUD2 marker. In both crosses and artificial fusions, the conversion of alpha- into alpha+ also influences the transmission of the more distant Nh, Nc and H markers. It is hypothesized that the more frequent transmission of the genome containing the intron results from the elimination of alpha- molecules, as a result of a double-strand cut which is induced by an endonuclease encoded by the intron.

Maternal transmission of mitochondrial DNA in interspecific hybrids of Populus.

Restriction fragment analysis was conducted to investigate the mode of inheritance of mitochondrial (mt) DNA in F1 progeny of two P. deltoides x P. deltoides, three P. deltoides x P. nigra, and two P. deltoides x P. maximowiczii controlled crosses, and in Populus x canadensis by using 16 restriction endonucleases and two heterologous probes of cloned mtDNA fragments of maize. Five restriction fragment length polymorphisms (RFLPs) of mtDNA differentiated P. deltoides from P. nigra, whereas three RFLPs of mtDNA separated P. deltoides from P. maximowiczii. In all cases, F1 progeny of P. deltoides x P. nigra, and P. deltoides x P. maximowiczii, crosses had mtDNA restriction fragments of only their maternal P. deltoides parents. P. x canadensis had mtDNA restriction fragments of only P. deltoides. F1 progeny of intraspecific P. deltoides crosses also had the same mtDNA fragments as their maternal parent. The results clearly demonstrate uniparental-maternal inheritance of the mitochondrial genome in F1 interspecific hybrids of P. deltoides with P. nigra and P. maximowiczii.

A nuclear mutation reversing a biased transmission of yeast mitochondrial DNA.

The highly biased transmission of p- mitochondrial DNA that occurs in hypersuppressive matings between p- and p+ cells of the yeast Saccharomyces cerevisiae is thought to be a consequence of the replication advantage of the p- mtDNA. A nuclear gene, MGT1, that is required for this displacement of p+ mtDNA from zygotic clones has been identified through mutation. When one haploid parent carries the mgt1 allele, transmission of p- mtDNA is substantially reduced. When both haploid parents carry the mgt1 allele, p- mtDNA is essentially eliminated from the zygotic progeny. Thus in the absence of the MGT1 gene there is a switch in the transmission bias; p+ mtDNA rather than the hypersuppressive p- mtDNA is inherited by most zygotic clones. In contrast to its semi-dominant behavior in haploid matings, mgt1 behaves as a recessive allele in diploid matings since the p+ genome in MGT1/mgt1 diploids is efficiently displaced when mated with a MGT1/mgt1 hypersuppressive p- diploid strain. We find that p+ genomes can be comaintained along with hypersuppressive p- mtDNA for extended periods in clonal lines derived from MGT1 x mgt1 matings. However, as expected from the recessive nature of the mgt1 mutation, these p+ genomes are eventually eliminated. Our work indicates that MGT1 plays a crucial role in the competition for inheritance between hypersuppressive p- mtDNAs and the p+ mitochondrial genome. The MGT1 gene product may be a component of a mtDNA replication system that acts preferentially at the rep sequences found in hypersuppressive mtDNAs.

Further study on selective transmission of mitochondrial DNA in heteroplasmic lines of Drosophila melanogaster.

The temperature-dependent transmission of mitochondrial DNA (mtDNA) was investigated in heteroplasmic lines of Drosophila melanogaster established by germ-plasm transplantation. Using D. melanogaster, D. simulans and D. mauritiana as germ-plasm donors, five recipient-donor combinations of heteroplasmy, differing from those previously examined (Matsuura et al., 1991), were constructed. For intraspecific reciprocal combinations, donor mtDNA in one combination was retained at 25 degrees C but was almost lost by the tenth generation at 19 degrees C. In the reciprocal, the proportion of the same type of recipient mtDNA decreased more quickly at 19 degrees C than 25 degrees C. Decreasing rates at 19 degrees C in the reciprocals differed from each other. For interspecific combinations, two species were used as germ-plasm donors. Donor mtDNA derived from D. simulans was lost at both temperatures and the rate of decrease was greater at 19 degrees C than 25 degrees C. The proportion of donor mtDNA derived from D. mauritiana increased at a greater rate at 25 degrees C than 19 degrees C when using two different strains of D. melanogaster as recipients. These results suggest that both the nuclear and two types of mitochondrial genomes are involved in the selective transmission of mtDNA.

Selective transmission of mitochondrial DNA in heteroplasmic lines for intra- and interspecific combinations in Drosophila melanogaster.

The transmission of mitochondrial DNA (mtDNA) was investigated in the heteroplasmic lines of Drosophila melanogaster at 19 degrees C and at 25 degrees C. The selective transmission of one type of mtDNA was dependent on the temperature at which the lines were maintained. In heteroplasmic lines for an intraspecific combination induced by germ-plasm transplantation using D. melanogaster as a germ-plasm donor, the proportion of donor mtDNA decreased in four out of five lines examined, the decreasing rate of which being greater at 25 degrees C than at 19 degrees C. Donor mtDNA was lost by the 20th generation at 25 degrees C. For an interspecific combination using D. mauritiana as a germ-plasm donor, the proportion of donor mtDNA increased and endogenous mtDNA was replaced with donor mtDNA at 25 degrees C. But donor mtDNA was almost lost at 19 degrees C by the 14th generation in all four lines examined. Possible mechanisms involved in the temperature-dependent modes of mtDNA transmission are discussed.

Selective transmission of mitochondrial DNA in Drosophila.

Selective transmission of mitochondrial DNA in Drosophila.

Incomplete maternal transmission of mitochondrial DNA in Drosophila.

The possibility of incomplete maternal transmission of mitochondrial DNA (mtDNA) in Drosophila, previously suggested by the presence of heteroplasmy, was examined by intra- and interspecific backcrosses of Drosophila simulans and its closest relative, Drosophila mauritiana. mtDNAs of offspring in these crosses were characterized by Southern hybridization with two alpha-32P-labeled probes that are specific to paternal mtDNAs. This method could detect as little as 0.03% paternal mtDNA, if present, in a sample. Among 331 lines that had been backcrossed for ten generations, four lines from the interspecific cross D. simulans (female) x D. mauritiana (male) showed clear evidence for paternal leakage of mtDNA. In three of these the maternal type was completely replaced while the fourth was heteroplasmic. Since in this experiment the total number of fertilization is known to be 331 x 10 = 3310, the proportion of paternal mtDNA per fertilization was estimated as about 0.1%. The mechanisms and evolutionary significance for paternal leakage are discussed in light of this finding.

Maternal transmission of mitochondrial DNA in ducks

Maternal transmission of mitochondrial DNA (mtDNA) has been studied in amphibians, insects and mammals, but little is known about mtDNA inheritance in the ovaripirous avian species. In this study, we have constructed the physical maps of mitochondrial genomes from two different genera of ducks ( Cairina and Anas ) and taken advantage of the availability of their hybrids to demonstrate that mtDNA is maternally inherited.

Conversion at large intergenic regions of mitochondrial DNA in Saccharomyces cerevisiae.

Saccharomyces cerevisiae mitochondrial DNA deletion mutants have been used to examine whether base-biased intergenic regions of the genome influence mitochondrial biogenesis. One strain (delta 5.0) lacks a 5-kilobase (kb) segment extending from the proline tRNA gene to the small rRNA gene that includes ori1, while a second strain (delta 3.7) is missing a 3.7-kb region between the genes for ATPase subunit 6 and glutamic acid tRNA that encompasses ori7 plus ori2. Growth of these strains on both fermentable and nonfermentable substrates does not differ from growth of the wild-type strain, indicating that the deletable regions of the genome do not play a direct role in the expression of mitochondrial genes. Examination of whether the 5- or 3.7-kb regions influence mitochondrial DNA transmission was undertaken by crossing strains and examining mitochondrial genotypes in zygotic colonies. In a cross between strain delta 5.0, harboring three active ori elements (ori2, ori3, and ori5), and strain delta 3.7, containing only two active ori elements (ori3 and ori5), there is a preferential recovery of the genome containing two active ori elements (37% of progeny) over that containing three active elements (20%). This unexpected result, suggesting that active ori elements do not influence transmission of respiratory-competent genomes, is interpreted to reflect a preferential conversion of the delta 5.0 genome to the wild type (41% of progeny). Supporting evidence for conversion over biased transmission is shown by preferential recovery of a nonparental genome in the progeny of a heterozygous cross in which both parental molecules can be identified by size polymorphisms.

Conversion at Large Intergenic Regions of Mitochondrial DNA in Saccharomyces cerevisiae

Saccharomyces cerevisiae mitochondrial DNA deletion mutants have been used to examine whether base-biased intergenic regions of the genome influence mitochondrial biogenesis. One strain (delta 5.0) lacks a 5-kilobase (kb) segment extending from the proline tRNA gene to the small rRNA gene that includes ori1, while a second strain (delta 3.7) is missing a 3.7-kb region between the genes for ATPase subunit 6 and glutamic acid tRNA that encompasses ori7 plus ori2. Growth of these strains on both fermentable and nonfermentable substrates does not differ from growth of the wild-type strain, indicating that the deletable regions of the genome do not play a direct role in the expression of mitochondrial genes. Examination of whether the 5- or 3.7-kb regions influence mitochondrial DNA transmission was undertaken by crossing strains and examining mitochondrial genotypes in zygotic colonies. In a cross between strain delta 5.0, harboring three active ori elements (ori2, ori3, and ori5), and strain delta 3.7, containing only two active ori elements (ori3 and ori5), there is a preferential recovery of the genome containing two active ori elements (37% of progeny) over that containing three active elements (20%). This unexpected result, suggesting that active ori elements do not influence transmission of respiratory-competent genomes, is interpreted to reflect a preferential conversion of the delta 5.0 genome to the wild type (41% of progeny). Supporting evidence for conversion over biased transmission is shown by preferential recovery of a nonparental genome in the progeny of a heterozygous cross in which both parental molecules can be identified by size polymorphisms.

Evidence for mitochondrial DNA polymorphism and uniparental inheritance in the cellular slime mold Polysphondylium pallidum: effect of intraspecies mating on mitochondrial DNA transmission.

Restriction fragment length polymorphisms (RFLPs) were used as markers to monitor mitochondrial inheritance in the cellular slime mold, Polysphondylium pallidum. When two opposite mating types (mat1 and mat2) of closely related strains were crossed, all the haploid progeny regardless of mating type inherited their mitochondrial DNA from the mat2 parent only. When opposite mating types from more distantly related strains were crossed, most of the progeny also inherited their mitochondrial DNA from the mat2 parent, but some inherited their mitochondrial DNA from the mat1 parent. In both cases however, the transmission of mitochondrial DNA was uniparental, since in every individual progeny only one type of mitochondrial DNA exists. Moreover, in crosses involving more distantly related strains all the progeny of a single macrocyst were shown to contain the same type of mitochondrial DNA. These findings are discussed in regard to mechanisms of transmission and the possible involvement of nuclear genes in the control of transmission of mitochondrial DNA in Polysphondylium.

Differences in the modes of transmission of foreign mitochondrial DNA in heteroplasmic lines for intra- and interspecific combinations in Drosophila melanogaster.

The transmission of foreign mitochondrial DNA (mtDNA) was investigated in heteroplasmic lines of Drosophila melanogaster constructed by germ-plasm transplantation and maintained at 19 degrees C. When D. melanogaster was used as a germ-plasm donor, donor mtDNA was retained in all four heteroplasmic lines examined. Individual females were found to be heteroplasmic at the 17th and 18th generations. Donor mtDNA derived from D. mauritiana was found to have decreased in all four heteroplasmic lines examined. It could no longer be found after the 16th generation. This difference in the modes of transmission of donor mtDNA in intra- and interspecific combinations of heteroplasmy indicates that there may be certain species-specific functions which propagate and transmit endogenous mtDNA under the nuclear genome of D. melanogaster.

Transmission of mitochondrial DNA in crosses involving diploid gametes homozygous or heterozygous for the mating-type locus in Chlamydomonas

The two interfertile algal species Chlamydomonas reinhardtii and C. smithii possess physically distinct mitochondrial (mit) genomes. Recently, use was made of this difference to demonstrate that sexual zygotes transmit the mit DNA from the mating-type minus (mt-, or paternal) parent exclusively. Diploid clones homozygous or heterozygous for the mt locus and carrying the mit genome of either of the two species were constructed by sexual crosses or artificially induced fusions. Haploid x diploid and diploid x diploid crosses were performed in order to analyze the role of both the mt locus and ploidy on the mode of transmission of mit DNA to the meiotic progeny. The inheritance of the mit DNA was determined by use of two molecular probes which hybridize to different regions of the organelle genomes. The mtu+/mt- gametes, which behave as mt- in the mating reaction, usually transmit their mit genome to the meiotic progeny, as do mt- or mt-/mt- gametes, regardless of the ploidy of the mt+ gametes. In the cross mt+x mt+/mt- however, 2 zygospore clones (out of 14) transmitted recombinant DNA molecules containing a large segment of the C. reinhardtii mit genome and a 1 kb fragment typical of C. smithii. It can thus be concluded that, contrary to what was observed earlier for chloroplast gene transmission: (1) mt- is dominant to mt+with regard to mit DNA transmission, and (2) nuclear ploidy has little, if any, effect on mit DNA transmission.