Inheritance of organelle DNA markers in a pea cross associated with nuclear-cytoplasmic incompatibility

An unusual biparental mode of plastid inheritance was found in pea, in a cross associated with nuclear-cytoplasmic incompatibility manifested as deficiency of chlorophyll pigmentation. Plastid DNA marker trnK and mitochondrial DNA marker cox1 were analyzed in F1 progeny that received cytoplasm from an accession of a wild subspecies Pisum sativum ssp. elatius. Plants with sectors of green tissue on leaves and seed cotyledons with green patches on an otherwise chlorotic background were found to carry paternally inherited plastid DNA, suggesting that photosynthetic function was affected by nuclear-cytoplasmic conflict and required proliferation of paternally inherited plastids for normal performance. The paternally inherited plastid DNA marker was also observed in the roots. The presence of the paternal marker in cotyledons, roots and leaves was independent of each other. Inheritance of the mitochondrial DNA marker cox1 appeared to be of the maternal type.     

苦苣苔科植物远缘杂交亲和性初探

该研究对苦苣苔科的蛛毛苣苔属[Paraboea(C.B.Clarke)Ridley]、马铃苣苔属(Oreocharis Bentham)、报春苣苔属(Primulina Hance)、吊石苣苔属(Lysionotus D.Don)、台闽苣苔属(Titanotrichum Solereder)、半蒴苣苔属(Hemiboea C.B.Clarke)、长蒴苣苔属(Didymocarpus Wallich)、光叶苣苔属(Glabrella Mich.MollerW.H.Chen)进行属间远缘杂交,对马铃苣苔属的马铃苣苔组、川滇马铃苣苔组、大叶石上莲组进行组间远缘杂交,统计不同杂交组合的结实情况及播种后的种子萌发情况,以明确不同属(组)间远缘杂交亲和性,为通过远缘杂交进行苦苣苔科种质资源的创新奠定基础。结果表明:(1)不同属作为父本,与同一母本进行杂交,以及同一属内不同种作为父本,与同一母本进行杂交,结实情况差异显著。(2)不同属作为母本,与同一父本进行杂交,以及同一属内不同种作为母本,与同一父本进行杂交,结实情况同样存在显著差异。(3)相同两个种正反交杂交成功率也并不一致。(4)马铃苣苔属与半蒴苣苔属,报春苣苔属与吊石苣苔属,吊石苣苔属与报春苣苔属和半蒴苣苔属,光叶苣苔属与报春苣苔属和吊石苣苔属之间部分种进行远缘杂交能够正常结实,且杂交种子能够正常萌发。(5)马铃苣苔属内组间杂交无明显生殖隔离的现象。     

Inheritance of organelle genomes in citrus somatic cybrids

Restriction fragment length polymorphisms (RFLPs) were used for the characterization of citrus organelle inheritance in somatic cybrids produced during six different citrus protoplast fusions. All the cybrids in this work inherited their mitochondrial genome from the embryogenic fusion partner (callus or cell suspension). In some of the combinations, non-parental bands were observed among the mitochondrial configurations. In contrast, the cybrids inherited plastid DNA from either the embryogenic or the nonembryogenic (leaf) fusion partner. The relative abundance of organelle DNAs in the embryogenic and leaf cells was in accordance with these inheritance patterns. Stochastic processes may therefore influence the outcome of somatic cell fusions with respect to organelle genomes.     

Molecular analysis of interspecific and intergeneric relationships of Banksia using RAPDs and non-coding chloroplast DNA sequences

 Random amplified polymorphic DNA (RAPD) and chloroplast DNA (cpDNA) markers were used to study interspecific and intergeneric relationships of Banksia (Proteaceae) to aid breeding of the genus for cut flower production. The accepted morphological phylogeny of Banksia, with two subgenera, two sections and 13 series, is unclear regarding the relationships of the commercial cut flower species B. coccinea. Fifteen RAPD primers and a non-coding cpDNA sequence between the trnL (UAA) and trnF (GAA) gene were applied to species of Banksia, the related genus Dryandra, and to Musgravea heterophylla as the outgroup, with cluster analysis applied to the results. The two methods were in broad agreement with each other, and with the accepted taxonomy, with closely related species pairs and groups clustering together, but RAPDs were not informative between distantly related species or species pairs. Banksia coccinea clustered with Dryandra and formed a polytomy with 2 Dryandra species and the two sections of subgenus Banksia. Subgenus Isostylis formed a polytomy with D. formosa, basal to subgenus Banksia, but with B. cuneata and B. illicifolia (both in subgenus Isostylis) polyphyletic. Dryandra did not separate as a clade and fell within Banksia, raising questions about the currently accepted view of the two as sister genera with parallel morphological development. The results indicate that interspecific and intergeneric hybridisation with genus Dryandra and subgenus Banksia may be possible routes for improvement of the commercial species B. coccinea. Received: 25 September 1996 / Accepted: 8 November 1996     

Nested clade analysis of Dunnia sinensis (Rubiaceae), a monotypic genus from China based on organelle DNA sequences

Dunnia sinensis, a monotypic genus of theRubiaceae endemic to the southeast mainlandChina, is an endangered species due to habitatdestruction over the past decades. Informationon levels and apportionment of geneticvariation across populations and geographicalregions is fundamental to conservation. In thepresent study, we used organelle DNA variationand nested phylogeographic analyses to test theisolation-by-distance model in this specieswith wind-mediated seed-dispersal and todistinguish ongoing gene flow from historicalprocesses. As expected, low levels of geneticvariation were detected at the ribosomal ITSregion of mtDNA ( = 0.0019± 0.0002) and the atpB-rbcLintergenic spacer of cpDNA ( =0.0022 ± 0.0009) in the rare species. Sixand seven haplotypes of mt- and cpDNA wereidentified from 125 individuals, respectively,according to the reconstructed neighbor-joiningtrees. Both data sets suggested consistentphylogenies that recovered two differentiatedlineages corresponding to western (Yangchun)and eastern (four others populations) portionsof the range. Hierarchical analyses of themolecular variance (AMOVA) of mt- and cpDNAindicated that molecular variance wasattributable to the difference between regions(ct = 0.911 and 0.771 for mt- and cpDNA,respectively) in D. sinensis. Based ongeographic distributions of haplotypes in thehaplotype networks, significant geneticdifferentiation between the two geographicregions, which can be seen as evolutionarilyconservation units, was associated withhistorical fragmentation. In contrast, limitedgene flow with occasional long-range dispersalshaped the apportionment of organelle DNAalleles among populations of the easternregion, within which two incompletely isolatedphylogeographic groups can be recognized asconservation units for management.     

Inheritance of organelle DNA in rye (Secale cereale L.) and wheat (x Triticale Thch.) hybrids

The mode of inheritance of chloroplast and mitochondrial DNA (mtDNA) in rye x triticale intergeneric hybrids has been studied with the use of specific PCR markers for loci 18S/5S and 3'rbcL in organelle DNA. In rye x triticale BC1, mtDNA copies of two types, paternal and maternal, have been found; in BC2 plants, only paternal mtDNA and chloroplast DNA (cpDNA) have been detected. Mechanisms determining the inheritance and/or differential amplification of organelles of a specific type are discussed.     

Molecular changes of organelle DNA sequences in rice through dedifferentiation,long-term culture,or the morphogenesis process

Callus-specific rearranged DNA in rice (Oryza sativa L.) was isolated by in-gel reassociation procedure. Southern hybridization experiments revealed that some clones were amplified significantly in primary callus induced from scutellum tissue. Rapid amplification of these clones was observed within 2 days after plating seeds onto callus-induction medium containing 2,4-D. NAA gave no significant effect on DNA amplification event. Colony formation process from isolated protoplasts and plant regeneration process from callus showed clone-specific and process-specific fluctuation patterns of copy number. Sequence analysis of the clones suggested that most of the clones were originated from organelle DNA. Comparison of copy number fluctuation pattern of organelle functional genes with that of the clones suggested multiformity and/or construction-specific amplification of organelle DNA.     

Inheritance of phenotypic traits in the progeny of a Ceratocystis interspecific cross

Ceratocystis fimbriata is a fungal plant pathogen that causes black rot on Ipomoea batatas. Based on inoculation studies on numerous tree species, the pathogen is known to be host specific. The closely related species, Ceratocystis manginecans, causes severe wilt on a broad range of tree hosts, including Mangifera indica, Acacia mangium and other leguminous tree species. The genetic factors underlying the pathogenicity and host specificity of Ceratocystis species have rarely been investigated. In this study, an F₁ population of 70 recombinant progeny from a cross between C. fimbriata and C. manginecans was generated and the inheritance of various phenotypic traits was investigated. Results showed that colony colour, growth rate, asexual spore production and aggressiveness to I. batatas and A. mangium are all quantitative traits with high levels of heritability. However, conidia production and aggressiveness appeared to be regulated by a small number of genes. No correlation could be found between aggressiveness and other phenotypic traits, suggesting that these are inherited independently. This is the first study to consider genetic inheritance of pathogenicity and host specificity in Ceratocystis species and the results will contribute, in future, to the identification of quantitative trait loci and candidate genes associated with the traits investigated.     

Inheritance of legumin and albumin contents in a cross between round and wrinkled peas

Summary Legumin and albumin are the fractions of pea seed proteins preferred to vicilin because of their high sulfur amino acid contents. The joint inheritance of legumin and albumin contents was studied in a cross between to contrasting lines of peas — one with high legumin and low albumin, and the other with low legumin and medium to high albumin. Single seed determinations were made in the parental, F₁; F₂ and backcross generations using rocket immunoelectrophoresis. In the non-segregating generations (P₁, P₂ and F₁), legumin and albumin contents were negatively correlated (r=–0.50). The estimates of correlation coefficients in the segregating generations (F₂, BC₁ and BC₂) were also about –0.5. However, the two estimates based on the round and on the wrinkled seeds separately in the F₂ generation were not significantly different from zero. At least four individual round F₂ seeds showed the desired recombination of high legumin with high albumin indicating that the unfavorable correlation can be broken. In this cross legumin content showed predominantly additive genetic variation whereas the dominance variance was the largest component for albumin content. A combined relative sulfur index, proposed as a convenient measure for selection, showed a narrow sense heritability of 47%. In general these results support the view that sulfur amino acid content of peas can be improved by breeding, but that the required selection regime must take both legumin and albumin content into account.     

Construction of a highly error-prone DNA polymerase for developing organelle mutation systems

A novel family of DNA polymerases replicates organelle genomes in a wide distribution of taxa encompassing plants and protozoans. Making error-prone mutator versions of gamma DNA polymerases revolutionised our understanding of animal mitochondrial genomes but similar advances have not been made for the organelle DNA polymerases present in plant mitochondria and chloroplasts. We tested the fidelities of error prone tobacco organelle DNA polymerases using a novel positive selection method involving replication of the phage lambda cI repressor gene. Unlike gamma DNA polymerases, ablation of 3′–5′ exonuclease function resulted in a modest 5–8-fold error rate increase. Combining exonuclease deficiency with a polymerisation domain substitution raised the organelle DNA polymerase error rate by 140-fold relative to the wild type enzyme. This high error rate compares favourably with error-rates of mutator versions of animal gamma DNA polymerases. The error prone organelle DNA polymerase introduced mutations at multiple locations ranging from two to seven sites in half of the mutant cI genes studied. Single base substitutions predominated including frequent A:A (template: dNMP) mispairings. High error rate and semi-dominance to the wild type enzyme in vitro make the error prone organelle DNA polymerase suitable for elevating mutation rates in chloroplasts and mitochondria.     

Sequence evidence for sporadic intergeneric DNA introgression from wheat into a wild Aegilops species

Introgressive hybridization has played a crucial role in the evolution of many plant species, especially polyploids. The duplicated genetic material and wide geographical distribution facilitate hybridization and introgression among polyploid species having either homologous or homoeologous genomes. Such introgression may lead to the production of recombinant genomes that are more difficult to form at the diploid level. Crop genes that have introgressed into wild relatives can increase the capability of the wild relatives to adapt to agricultural environments and compete with crops or to compete with other wild species. Although the transfer of genes from crops into their conspecific immediate wild progenitors has been reported, little is known about spontaneous gene movement from crops to more distantly related species. We describe recent spontaneous DNA introgression from domesticated polyploid wheat into distantly related, wild tetraploid Aegilops peregrina (syn. Aegilops variabilis) and the stabilization of this sequence in wild populations despite not having homologous chromosomes. Our results show that DNA can spontaneously introgress between homoeologous genomes of species of the tribe Triticeae and, in the case of crop-wild relatives, possibly enrich the wild population. These results also emphasize the need for fail-safe mechanisms in transgenic crops to prevent gene flow where there may be ecological risks.     

Inheritance of seed desiccation sensitivity in a coffee interspecific cross: evidence for polygenic determinism

The genetic determinism of seed desiccation sensitivity was studied using a cross between two coffee species exhibiting a large difference for this trait, Coffea pseudozanguebariae (tolerant) and C. liberica (sensitive). Throughout the whole study, seed desiccation tolerance was quantified both in terms of water content and water activity. Whatever the parameter used, the level of seed desiccation tolerance in F1 hybrids corresponded to that of the mid-parent, thus indicating an additive inheritance of seed desiccation tolerance at the F1 level. A broad variation was observed among hybrids backcrossed to C. liberica (BCs) for seed desiccation tolerance, independent of the parameter used to quantify it. This variation was continuous and BCs showed transgression in the direction of the most desiccation sensitive parent, indicating (i) that desiccation tolerance is a polygenic trait in coffee species, and (ii) that C. pseudozanguebariae does not present the most favourable alleles for all the genes involved in seed desiccation tolerance. No significant difference was observed between the two reciprocal backcrosses, F1xC. liberica and C. libericaxF1, for the level of desiccation tolerance of their seeds, showing the absence of a maternal effect on this trait. There was no significant effect of the number of seeds harvested from each BC on the level of desiccation tolerance of its seeds. Moreover, there was no significant correlation within BCs between seed size, seed viability, and water content before desiccation and desiccation tolerance.     

Inheritance of chloroplast DNA inPopulus

Summary Restriction fragment length polymorphisms (RFLPs) were used as markers to determine the transmission of chloroplast DNA (cpDNA) in poplar crosses. The plant material studied included individual trees ofPopulus trichocarpa, P. maximowiczii xtrichocarpa, P. maximowiczii xnigra, and offspring from controlled crosses between these trees. RFLPs were identified by direct observation of stained restriction fragments, as well as by molecular hybridization with heterologous cpDNA probes. Analysis of the restriction fragment patterns in the parents and their progeny showed only the patterns of the maternal tree in the progeny, while no paternal type was found. These results provide clear evidence of a maternal mode of chloroplast inheritance in the poplar clones studied.     

A genetic map of citrus based on the segregation of isozymes and RFLPs in an intergeneric cross

Summary Isozymes and restriction fragment length polymorphisms were used as markers in the construction of a genetic map of the citrus nuclear genome. The map was based on the segregation of 8 isozyme, 1 protein, and 37 RFLP loci in 60 progeny of a cross of two intergeneric hybrids, Sacaton citrumelo (Citrus paradisi Macf. x Poncirus trifoliata (L.) Raf.) and Troyer citrange (C. sinensis (L.) Osbeck x P. trifoliata), often used as rootstocks. The map contains 38 of 46 studied loci distributed on ten linkage groups. A genome size of 1,700 cM was estimated from partial linkage data. Approximately 35% of the genome should be within 10 cM and 58% within 20 cM of the mapped markers. Eight loci in three linkage groups and 1 unlinked locus deviated significantly from Mendelian segregation.     

Tissue-specific organelle DNA degradation mediated by DPD1 exonuclease

Organelle DNA in plastids and mitochondria is present in multiple copies and undergoes degradation developmentally. For example, organelle DNA that is detectable cytologically using DNA-fluorescent dye disappears during pollen development. Nevertheless, nucleases involved in this degradation process remain unknown. Our recent study identified the organelle nuclease, DPD1, which has Mg²⁺-dependent exonuclease activity in vitro. The discovery of DPD1 emerged from Arabidopsis mutant screening and concomitant isolation of dpd1 mutants that retain organelle DNA in mature pollen. DPD1 is conserved only in angiosperms: not in other photosynthetic organisms. Despite these findings, the physiological significance of organelle DNA degradation during pollen development remains unclear because dpd1 exhibits no apparent defects in pollen viability or in the maternal inheritance of organelle DNA. We discuss a possible role of organelle DNA degradation mediated by DPD1, based on a DPD1 expression profile studied using in silico analyses.Key words: mitochondria, nuclease, organelle DNAs, plastids, pollen