Genes in mitochondria and chloroplasts behave quite differently from genes in the nucleus: they are often inherited from only one parent, and they segregate during mitotic cell divisions. Cells con...

https://science.sciencemag.org/content/sci/222/4623/468.full.pdf

Relaxed cellular controls and organelle heredity

A mating type-linked gene cluster expressed in Chlamydomonas zygotes participates in the uniparental inheritance of the chloroplast genome

In vivo complementation between different wild and mutant strains defective for nitrate assimilation has been performed by isolating diploid strains from the appropriate crosses. Twenty-two diploids homozygous or heterozygous with respect to nitrate reduction and able to grow on nitrate medium were obtained and their diploid character demonstrated from analyses of mating type, cell volume, nuclear size and progeny of crosses with haploid wild-type. All diploids were assayed for overall- and terminal-nitrate reductase (NR) activity and for the occurrence of the NR-diaphorase subunit. Data on NR activities in heterozygotes carrying mutation(s) in structural gene(s) (nit-1 or nit-1a, nit-1b) agree with the heteromultimeric nature of the enzyme complex previously described (Franco et al. (1984) EMBO J 3: 1403–1407), and indicate that subunits are exchangeable to form hybrid enzymes. In addition, in vitro complementation tests with mutant nit-1 of C. reinhardtii indicate that this mutant has defective NR-diaphorase subunits but intact terminal subunits. Super-repression caused by the mutant allele nit-2 is suppressed by the wild allele in heterozygotes, which suggests a positive control by the nit-2 product on structural gene(s) transcription. Mutant alleles of genes for the biosynthesis of molybdenum-containing cofactor, either nit-4 or nit-5 and nit-6, were recessive in diploids carrying them. The mutant allele of nit-3, from strain 307, was codominant in all heterozygotes suggesting that nit-3 codes for a protein whose activity is limiting for the molybdenum-cofactor biosynthetic pathway.

In vivo complementation analysis of nitrate reductase-deficient mutants in Chlamydomonas reinhardtii.

The biparental transmission of the mitochondrial genome in chlamydomonas reinhardtii visualized in living cells

Expression of three chloroplast genes encoding proteins of different chloroplast complexes and the rRNA gene has been examined in cells having reduced numbers of chloroplast genomes as a result of growth in the presence of the thymidine analog 5-fluorodeoxyuridine. While accumulation of total mRNA for rpl2 (ribosomal protein L-1), rbcL (ribulose bisphosphate carboxylase large subunit) and atpA (alpha-subunit of ATP synthase) declined with gene copy number, the levels of translatable mRNA and rates of synthesis of these three proteins were largely unaffected. Accumulation of rRNA declined less precipitously than mRNA levels for the three proteins in response to the reduction in chloroplast genome number. Chlamydomonas appears to compensate for reductions in the number of chloroplast genomes at several different levels. Populations of cells with only one-fourth the wild-type amount of chloroplast DNA per cell on average have half the normal level of chloroplast ribosomes and nearly normal rates of CO(2) fixation and levels of specific chloroplast encoded proteins. These results suggest that normal cells accumulate a large excess of transcripts for chloroplast genes and that levels of expression of these genes are regulated by posttranscriptional mechanisms.

Relationship between Gene Dosage and Gene Expression in the Chloroplast of Chlamydomonas reinhardtii

Somatic fusion between strains of Chlamydomonas containing complementing cell-wall and auxotrophic mutations, having the same mating-type (mt) and bearing chloroplast markers, have been performed to study the mode of chloroplast gene inheritance in the fusion products. About one third of the fusion products (mt
+/mt
+ or mt
−/mt
−) transmitted chloroplast markers from both parents (= biparental fusion products). The rest of the population was equally distributed between fusion products transmitting the chloroplast marker of one parent or the other (uniparental fusion products) exclusively. Incubation of the fusion products in the dark for 48 hours, immediately after the fusion, decreases the frequency of biparental fusion products. The results indicate that the general process of elimination of chloroplast alleles is independent of the presence of both mt
+ and mt
− alleles in the cell. In contrast, directional elimination (i.e. preferential elimination of paternal chloroplast alleles) does appear to depend upon heterozygosity at the mt locus. These results are discussed in relation to the models which have been proposed to explain the maternal inheritance of chloroplast genes in Chlamydomonas.

Chloroplast gene inheritance studied by somatic fusion in Chlamydomonas reinhardtii.

Sexual crosses and somatic fusions were performed between complementing wall-less arg- mutant strains bearing chloroplast markers for resistance to antibiotics. The mode of chloroplast allele transmission was investigated in the diploid colonies developed from both vegetative zygotes and fusion products. Before mating or fusion, one or both of the parental strains were grown for 4 or 8 days on agar containing 5-fluorodeoxyuridine (FUdR, 0.1 to 1.0 mM), which selectively reduces the amount of chloroplast DNA in Chlamydomonas. When one parent was pregrown on FUdR, the frequency of vegetative zygotes transmitting chloroplast alleles of both parents (biparental or BP zygotes) decreased, the reduction being more drastic when the mt parent was treated. Transmission was mainly uniparental maternal (UPm) or paternal (UPp) depending on whether the mt- or the mt+ parent was pregrown for 8 days in the presence of 1 .O mM FUdR. Treatment of both parents led to a strong maternal transmission. In the experiments involving somatic fusion between parent 1 and parent 2 (same or opposite mt), the ratio UP1/UP2, which was approximately equal to 1 in the control, decreased or increased according to whether the cells of parent 1 or 2 were pregrown on FUdR. In parallel, the frequency of BP fusion products always decreased. When both parental strains were treated with FUdR, the frequency of BP fusion products also decreased and the ratio UP,/cTP, was roughly equal to 1. The effect of FUdR can be interpreted in terms of reduction of the input frequencies of parental chloroplast genomes at the time of gametic or somatic cell fusion, the bias in favor of the maternal parent being operational only in sexual crosses.

https://www.genetics.org/content/genetics/99/3-4/371.full.pdf

Marie-Paule Hermesse

Papers

Chloroplast gene inheritance studied by somatic fusion in Chlamydomonas reinhardtii.

Modification of chloroplast gene transmission in somatic fusion products and vegetative zygotes of chlamydomonas reinhardi by 5-fluorodeoxyuridine