Heteroplasmy and paternally oriented shift of the organellar DNA composition in barley–wheat hybrids during backcrosses with wheat parents

Genome ◽  
2005 ◽  
Vol 48 (5) ◽  
pp. 761-769 ◽  
Author(s):  
Elena Aksyonova ◽  
Marina Sinyavskaya ◽  
Nina Danilenko ◽  
Lidia Pershina ◽  
Chiharu Nakamura ◽  
...  
Keyword(s):  
Simple Sequence Repeat Locus ◽  
Nuclear Genome ◽  
Upstream Region ◽  
Male Sterile ◽  
Plant Vigour ◽  
Organelle Dna ◽  
Advanced Backcross ◽  
Organellar Dna ◽  
Flanking Region ◽  
Wheat Hybrids

Mitochondrial (mt) and chloroplast (ct) genome inheritance was studied in barley–wheat hybrids, as were their progenies obtained from backcrosses with different common wheat cultivars, by monitoring the composition of 4 mtDNA (coxI, a 5′-flanking region of cob, nad3-orf156, and 5′-upstream region of 18S/5S) and 2 ctDNA (simple-sequence repeat locus downstream of trnS and a 3′-flanking region of rbcL) loci. In male sterile F1 and BC1 plants, maternal barley mtDNA fragments were mainly detected and very low levels of paternal wheat fragments were occasionally detected by PCR in coxI, a 5′-flanking region of cob and nad3-orf156, whereas a 5′-upstream region of 18S/5S showed clear heteroplasmy, containing both maternal and paternal copies, with maternal copies prevailing. Plants showing such heteroplasmic mtDNA composition remained either semisterile or became completely sterile in the later backcross generations. Only maternal ctDNA copies were detected in these plants. In 3 stable, self-fertile, and vigourous lines obtained in the advanced backcross generations and possessing recombinant wheat nuclear genome, however, only mt- and ctDNA copies of wheat parents were detected; thus, the original alloplasmic condition appeared to be lost. Our results suggest that transmission followed by selective replication of the paternal wheat organellar DNA leads to a paternally oriented shift of the organellar DNA composition in barley–wheat hybrids, which correlates with the restoration of fertility and plant vigour. These 2 processes seem to be related to nucleocytoplasmic compatibility and to be under the control of the nuclear genome composition.Key words: barley–wheat hybrids, ctDNA, mtDNA, nucleoytoplasmic compatibility, organelle DNA inheritance.

Structural Analysis and Identification of Cis-Elements of Rice osRACD Gene

2004 ◽  
Vol 36 (3) ◽  
pp. 191-198 ◽  
Author(s):  
Yun-Zhe Xu ◽  
Rui-Lin You ◽  
Nai-Hu Wu
Keyword(s):  
Oryza Sativa L ◽  
Small Gtpase ◽  
Inverse Pcr ◽  
Upstream Region ◽  
Male Sterile ◽  
Mobility Shift ◽  
Nuclear Extracts ◽  
Mobility Shift Assay ◽  
Photoperiod Sensitive ◽  
Gel Mobility Shift

Abstract The osRACD gene correlated with fertility transformation in the photoperiod sensitive genic male sterile rice (PGMR), Nongken 58S, encoded a rice (Oryza sativa L. ssp. japonica) small GTPase belonging to the Rac/Rho family. Inverse PCR was performed to amplify a fragment about 1.4 kb in 5′ upstream region of the osRACD promoter. Deletion mutation and gel mobility shift assay characterized two fragments (–799 to –686 nt, and –686 to –431 nt) in the osRACD promoter that could be involved in its transcriptional regulation. When these two deletion fragments were used as probe respectively, a retarded band appeared in the nuclear extracts of fertile 58S rice under short day (58S-SD). Whereas no retarded band was shown in the nuclear extracts of sterile 58S rice under long day (58S-LD). Competition assay indicated that the factors in the retarded bands binding to these two fragments were the same trans-acting factor (termed rice factor, RF). The binding affinity of RF was affected by phosphorylation and was higher in SD-growth rice than that of LD-growth rice.

scholarly journals Genomic footprinting of a yeast tRNA gene reveals stable complexes over the 5'-flanking region.

1989 ◽  
Vol 9 (8) ◽  
pp. 3244-3252 ◽  
Author(s):  
J M Huibregtse ◽  
D R Engelke
Keyword(s):  
Stationary Phase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Dnase I ◽  
Transcription Initiation Site ◽  
Upstream Region ◽  
Stable Complex ◽  
Coding Region ◽  
Flanking Region ◽  
Genomic Footprinting

We have shown by genomic footprinting that the 5'-flanking region of the Saccharomyces cerevisiae tRNASUP53 gene is protected from DNase I digestion. The protected region has a 5' boundary at -40 (relative to the transcription initiation site) and extends into the coding region of the gene, with a 3' boundary at approximately +15. Although the DNase I protection over this region was much greater than at the A- and B-box internal promoters, point mutations within the A or B box that reduced transcription in vitro eliminated the upstream DNase I protection. This implies that formation of a stable complex over the 5'-flanking region is dependent on interaction of the gene with transcription factor IIIC but that stability of the complex may not require continued interaction with this factor. The DNase I protection under varied growth conditions further suggested that the upstream complex is composed of two or more components. The region over the transcription initiation site (approximately +15 to -10) was less protected in stationary-phase cultures, whereas the more upstream region (approximately -10 to -40) was protected in both exponential- and stationary-phase cultures.

scholarly journals Development and Fertility Restoration of CMS Eggplant Lines Carrying the Cytoplasm of Solanum violaceum

2016 ◽  
Vol 8 (2) ◽  
pp. 10
Author(s):  
Konstantinos S. Krommydas ◽  
Zisis Tzikalios ◽  
Panagiotis Madesis ◽  
Fotios A. Bletsos ◽  
Athanasios Mavromatis ◽  
...  
Keyword(s):  
Male Fertility ◽  
Fertility Restoration ◽  
Genetic Locus ◽  
Pollen Viability ◽  
Male Sterile ◽  
Mt Dna ◽  
Normal Morphology ◽  
Advanced Backcross ◽  
The Past ◽  
Backcross Populations

<p>A functional cytoplasmic male sterile (CMS) eggplant line carrying the cytoplasm of <em>Solanum violaceum</em> was developed in the past, but the fertility restoring genes (<em>Rf</em>-genes) were not identified. This work aimed to produce the CMS lines of three Hellenic eggplant cultivars (viz., ‘Langada’, ‘Emi’ and ‘Tsakoniki’) using the cytoplasm of <em>S. violaceum</em> and study the inheritance of the <em>Rf</em>-genes. The respective CMS eggplant lines were developed by the backcross method and examined for their fertility parameters. The results demonstrated that female fertility was not affected by the cytoplasm of <em>S. violaceum</em>. In contrast, the occurrence of three male fertility phenotypes (male sterile, male fertile and potentially male fertile) indicated that male fertility was affected by nuclear/cytoplasmic interactions. Male sterile plants were characterized by indehiscent anthers, low pollen viability and abnormal anther morphology. Male fertile plants formed dehiscent anthers with high pollen viability and normal morphology. Potentially male fertile plants initially formed dehiscent anthers, but in later stages formed exclusively indehiscent anthers. Male fertile plants were obtained in the advanced backcross populations of CMS ‘Tsakoniki’, but not in CMS ‘Langada’ and CMS ‘Emi’. The genetic analysis of fertility restoration indicated that male fertility in the genetic background of cv. ‘Tsakoniki’ is controlled by one essential genetic locus, affected by a secondary modifying locus. Molecular analysis of cp-DNA and mt-DNA in the CMS lines indicated maternal inheritance of the cytoplasm organelles. Our findings demonstrate that the genotype of the eggplant parent can affect the expression of CMS as well as fertility restoration.</p>

Genomic footprinting of a yeast tRNA gene reveals stable complexes over the 5'-flanking region

1989 ◽  
Vol 9 (8) ◽  
pp. 3244-3252
Author(s):  
J M Huibregtse ◽  
D R Engelke
Keyword(s):  
Stationary Phase ◽  
Transcription Initiation ◽  
Initiation Site ◽  
Dnase I ◽  
Transcription Initiation Site ◽  
Upstream Region ◽  
Stable Complex ◽  
Coding Region ◽  
Flanking Region ◽  
Genomic Footprinting

We have shown by genomic footprinting that the 5'-flanking region of the Saccharomyces cerevisiae tRNASUP53 gene is protected from DNase I digestion. The protected region has a 5' boundary at -40 (relative to the transcription initiation site) and extends into the coding region of the gene, with a 3' boundary at approximately +15. Although the DNase I protection over this region was much greater than at the A- and B-box internal promoters, point mutations within the A or B box that reduced transcription in vitro eliminated the upstream DNase I protection. This implies that formation of a stable complex over the 5'-flanking region is dependent on interaction of the gene with transcription factor IIIC but that stability of the complex may not require continued interaction with this factor. The DNase I protection under varied growth conditions further suggested that the upstream complex is composed of two or more components. The region over the transcription initiation site (approximately +15 to -10) was less protected in stationary-phase cultures, whereas the more upstream region (approximately -10 to -40) was protected in both exponential- and stationary-phase cultures.

Delimitation and characterization of cis-acting DNA sequences required for the regulated expression and transcriptional control of the chicken skeletal alpha-actin gene

1986 ◽  
Vol 6 (7) ◽  
pp. 2462-2475
Author(s):  
D J Bergsma ◽  
J M Grichnik ◽  
L M Gossett ◽  
R J Schwartz
Keyword(s):  
Dna Sequences ◽  
Transcriptional Control ◽  
Regulatory Element ◽  
Upstream Region ◽  
Xenopus Laevis Oocytes ◽  
Actin Gene ◽  
Base Pairs ◽  
Cis Acting ◽  
Flanking Region ◽  
Alpha Actin

We have previously observed that DNA sequences within the 5'-flanking region of the chicken skeletal alpha-actin gene harbor a cis-acting regulatory element that influences cell type and developmental stage-specific expression (J. M. Grichnik, D. J. Bergsma, and R. J. Schwartz, Nucleic Acids Res 14:1683-1701, 1986). In this report we have constructed unidirectional 5'-deletion and region-specific deletion-insertion mutations of the chicken skeletal alpha-actin upstream region and inserted these into the chloramphenicol acetyltransferase expression vector pSV0CAT. These constructions were used to locate DNA sequences that are required for developmental modulation of expression when transfected into differentiating myoblasts. With this assay we have delimited the 5' boundary of a cis-acting regulatory element to ca. 200 base pairs upstream of the mRNA cap site. In addition, we have preliminarily identified DNA sequences that may be important subcomponents within this element. A second major focus of this study was to identify those DNA signals within the regulatory element that control transcription. Toward this end, the expression phenotypes of progressive 5'-deletion and deletion-insertion mutants of the 5'-flanking region of the chicken skeletal alpha-actin gene were assayed in microinjected Xenopus laevis oocytes. These experiments defined a cis-acting transcriptional control region having a 5' border 107 base pairs preceding the alpha-actin RNA cap site. Proximal and distal functionally important regions of DNA were identified within this element. These DNA signals included within their DNA sequences the "CCAAT" and "TATA" box homologies.

scholarly journals Cloning and characterization of the 5′-flanking region of the oxalate decarboxylase gene from Flammulina velutipes

2002 ◽  
Vol 367 (1) ◽  
pp. 67-75 ◽  
Author(s):  
Mohammad AZAM ◽  
Meenu KESARWANI ◽  
Subhra CHAKRABORTY ◽  
Krishnamurthy NATARAJAN ◽  
Asis DATTA
Keyword(s):  
Transcription Factor ◽  
Oxalic Acid ◽  
Specific Binding ◽  
Low Ph ◽  
Flammulina Velutipes ◽  
Upstream Region ◽  
Oxalate Decarboxylase ◽  
Mobility Shift ◽  
Cell Extracts ◽  
Flanking Region

The oxalate-degrading enzyme, oxalate decarboxylase (OXDC), was purified and characterized from Flammulina velutipes, a basidiomycetous fungus [Mehta and Datta (1991) J. Biol. Chem. 266, 23548—23553]. The cDNA cloning and analyses revealed that OXDC transcription was induced by oxalic acid. However, in this report, we show that OXDC transcription is induced by low pH, not by oxalate. To understand the regulatory mechanism of OXDC expression, we have cloned and analysed a 580-bp genomic fragment from the 5′-flanking region of the OXDC gene. Sequence analysis showed the presence of several eukaryotic transcription factor binding motifs within the −580bp of the upstream region. Electrophoretic-mobility-shift assays with partially purified cell extracts revealed specific binding of a factor in acid-induced, but not in uninduced, extracts. Furthermore, DNase I protection assays using the partially purified fraction from oxalic acid-induced extract revealed a footprint of a 13-bp sequence 5′GCGGGGTCGCCGA3′, termed low pH responsive element (LPRE), corresponding to the −287 to −275bp region of the OXDC promoter. Our results suggest that in F. velutipes cells, activation of OXDC transcription in response to low pH is mediated by the binding of a novel transcription factor through the LPRE site in the OXDC promoter.

High-frequency embryogenesis in male sterile plants of Brassica napus through microspore culture

1988 ◽  
Vol 66 (8) ◽  
pp. 1676-1680 ◽  
Author(s):  
Phan V. Chuong ◽  
K. P. Pauls ◽  
W. D. Beversdorf
Keyword(s):  
Brassica Napus ◽  
Male Sterility ◽  
Constant Temperature ◽  
Oilseed Rape ◽  
High Frequency ◽  
Microspore Culture ◽  
Doubled Haploids ◽  
Nuclear Genome ◽  
Male Sterile ◽  
Cytoplasmic Male Sterile

Microspores from several Polima cytoplasmic male sterile (Pol-CMS) and Diplotaxis muralis male sterile (Mur-MS) oilseed rape lines were cultured in a modified Nitsch and Nitsch medium in the dark for 4 weeks. High androgenic frequencies were observed in microspore cultures initiated from plants of both male sterile (MS) types. In cultures maintained at a constant temperature (30 °C) 1 or 2% of the microspores from Pol-CMS or Mur-MS lines, respectively, developed into embryos. A greater percentage of the embryos obtained from the Pol-CMS lines were of good quality than those obtained from the Mur-MS lines (25 vs. 5%). Twenty percent of the plants regenerated from embryos of both lines were spontaneous doubled haploids. The results suggest that no unfavorable cytoplasmic–nuclear genome interaction affecting androgenic potential exists in plants exhibiting Polima or Diplotaxis muralis male sterility.

scholarly journals Experimental evolutionary studies on the genetic autonomy of the cytoplasmic genome “plasmon” in the Triticum (wheat)–Aegilops complex

2019 ◽  
Vol 116 (8) ◽  
pp. 3082-3090 ◽  
Author(s):  
Koichiro Tsunewaki ◽  
Naoki Mori ◽  
Shigeo Takumi
Keyword(s):  
Maternal Inheritance ◽  
Nuclear Genome ◽  
Genetic Effect ◽  
Genetic System ◽  
Critical Examination ◽  
Organellar Dna ◽  
Aegilops Caudata ◽  
The Relationship ◽  
Simple Sequence ◽  
Dna Characteristics

The term “plasmon” is used to indicate the whole cytoplasmic genetic system, whereas “genome” refers to the whole nuclear genetic system. Although maternal inheritance of the plasmon is well documented in angiosperms, its genetic autonomy from the coexisting nuclear genome still awaits critical examination. We tested this autonomy in two related studies: One was to determine the persistence of the genetic effect of the plasmon of Aegilops caudata (genome CC) on the phenotype of common wheat, Triticum aestivum strain “Tve” (genome AABBDD), during 63 y (one generation per year) of repeated backcrosses of Ae. caudata and its offspring with pollen of the same Tve wheat, and the second was to reconstruct an Ae. caudata strain from the genome of this strain and its plasmon that had been resident in Tve wheat for 50 generations, and to compare the phenotypic and organellar DNA characteristics between the native and reconstructed strains. Results indicated no change in the effect of Ae. caudata plasmon on Tve wheat during its stay in wheat for more than half a century, and no difference between the native and reconstructed caudata strains in their phenotype and simple sequence repeats in their organellar DNAs, thus demonstrating the prolonged genetic autonomy of the plasmon from the coexisting genomes of wheat and several other species that were used in the reconstruction of Ae. caudata. The relationship between the proven genetic autonomy of the plasmon under changing nuclear conditions and its diversification during evolution of the Triticum–Aegilops complex is discussed.

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