Inferring the history of speciation in house mice from autosomal,X‐linked,Y‐linked and mitochondrial genes

Patterns of genetic differentiation among taxa at early stages of divergence provide an opportunity to make inferences about the history of speciation. Here, we conduct a survey of DNA‐sequence polymorphism and divergence at loci on the autosomes, X chromosome, Y chromosome and mitochondrial DNA in samples of Mus domesticus, M. musculus and M. castaneus. We analyzed our data under a divergence with gene flow model and estimate that the effective population size of M. castaneus is 200 000–400 000, of M. domesticus is 100 000–200 000 and of M. musculus is 60 000–120 000. These data also suggest that these species started to diverge approximately 500 000 years ago. Consistent with this recent divergence, we observed considerable variation in the genealogical patterns among loci. For some loci, all alleles within each species formed a monophyletic group, while at other loci, species were intermingled on the phylogeny of alleles. This intermingling probably reflects both incomplete lineage sorting and gene flow after divergence. Likelihood ratio tests rejected a strict allopatric model with no gene flow in comparisons between each pair of species. Gene flow was asymmetric: no gene flow was detected into M. domesticus, while significant gene flow was detected into both M. castaneus and M. musculus. Finally, most of the gene flow occurred at autosomal loci, resulting in a significantly higher ratio of fixed differences to polymorphisms at the X and Y chromosomes relative to autosomes in some comparisons, or just the X chromosome in others, emphasizing the important role of the sex chromosomes in general and the X chromosome in particular in speciation.     

Isolation of MTX-resistant Cell Line NP-19 of Nicotiana plumbaginifolia: Phenotypic, Genetic and Biochemical Study

Several cell lines of Nicotiana plumbaginifolia resistant tomethotrexate (MTX) were isolated upon gradual elevation of theMTX concentration in the growth medium. One of the MTX-resistantcell lines, NP-19, acquired resistance to 10 mmol m^–3MTX, that is, at least 50-fold higher than the lethal dose forthe wild-type cell suspension. Its resistance was stably maintainedupon prolonged withdrawal of the drug. The acquisition of resistancewas accompanied by severe deterioration of regeneration potential.Regenerated shoots still manifested resistance to 50 mmol m^–3.MTX-resistant colonies and tiny shoots were recovered followingasymmetric fusion between -irradiated NP-19 protoplasts andN. plumbaginifolia leaf mesophyll-derived protoplasts on a mediumcontaining 10 mmol m^–3 MTX. The development of the shootsderived both from NP-19 calli and following somatic fusion wasarrested at the stage of 6–8 leaves. No difference wasfound in uptake of ³H-MTX between cell suspension of NP-19 andthe parental cell line. A 30-fold increase in binding of ³H-MTXto protein extract was found in cell line NP-19, suggestingdifferential capacity of MTX binding as a mechanism involvedin the MTX resistance of this cell line. Since differentiatedorgans seem more sensitive to MTX than undifferentiated tissues,this cell line is a promising source for a gene(s) conferringenhanced MTX-tolerance both in non-differentiated and differentiatedtissues. Key words: Methotrexate resistance, MTX-binding capacity, MTX uptake, tobacco cell culture     

Organs and Plantlets Regeneration of Gladiolus through Tissue Culture

Explants from inflorescence stalks of Gladiolus when culturedin vitro regenerated new plantlets within 6–7 weeks. Regenerationwas started by the formation on the basal end of a thin layerof callus and root primordia. This was followed by formationof buds and cormlets, on the distal end. The regeneration ofthe various organs from the explants was found to be polarizedand depended on the levels of growth substances added to thebasal medium, best combination for organ initiation being 10ppm naphthalene-acetic acid and 0.5 ppm of kinetin.     

Competition of Ethionine and Methionine for Aminoacyl-tRNA Formation in an In Vitro System from Ochromonas danica*

Aminoacyl-tRNA synthetase and tRNA were isolated from the chrysomonad Ochromonas danica. The mutual effect of methionine and ethionine, and the effect of other amino acids on methionyl- and ethionyl-tRNA formation, were tested in an in vitro system. The tRNA^Met had a similar accepting capacity for either methionine or ethionine. Ethionine and methionine, but none of the other amino acids tested, competed for the same aminoacyl-tRNA synthetase. The K_m of methionine was 0.88 × 10^–5 M, and that of ethionine 5 × 10^–4 M. Ethionine inhibited methionine binding; K_i 3.4 × 10^–4 M. The respective values in a similar system isolated from E. coli were 2.2 × 10^–5, 1.95 × 10^–3, and 1.95 × 10^–3.     

Geotropic Responses and Pod Development in Gynophore Explants of Peanut (Arachis hypogaea L.) Cultured In Vitro

Peanut gynophore explants cultured in vitro on a defined mediumshow a positive geotropic response in both light and dark whenplanted either horizontally, or vertically with the tip pointingupwards. The growth following the initial curvature dependedon age of the gynophores and on the levels of growth substancesin the medium. In the dark and in presence of 0·01–0·1p.p.m. kinetin, naphthalene acetic acid at concentrations of0·1 p.p.m. and lower promoted gynophore elongation. Athigher concentrations elongation was promoted to a lesser extentin younger explants, caused enlargement of the ovary and formationof pods. Young explants generally elongated more than olderones and pod formation took place inside the medium, while inolder ones it took place above the medium. In the light, theinitial positive geotropic response was followed by elongationbut without any enlargement of the ovary. Decapitation of gynophores1·5–2·0 mm below their tip, removing theovary but leaving most of the intercalary meristem, had no effecton the geotropic response and elongation. The initial geotropicresponse and elongations of explants in vitro was not dependenton the presence of the ovary but on the meristem proximal toit. Changes in growth substances balance during gynophore developmentseem to affect geotropic response, elongation and pod formationin the peanut.     

The global nitrogen regulator, FNR1, regulates fungal nutrition-genes and fitness during Fusarium oxysporum pathogenesis

Fusarium oxysporum is a soil-borne pathogen that infects plants through the roots and uses the vascular system for host ingress. Specialized for this route of infection, F. oxysporum is able to adapt to the scarce nutrient environment in the xylem vessels. Here we report the cloning of the F. oxysporum global nitrogen regulator, Fnr1 , and show that it is one of the determinants for fungal fitness during in planta growth. The Fnr1 gene has a single conserved GATA-type zinc finger domain and is 96% and 48% identical to AREA-GF from Gibberella fujikuroi , and NIT2 from Neurospora crassa , respectively. Fnr1 cDNA, expressed under a constitutive promoter, was able to complement functionally an N. crassa nit-2 ^RIP mutant, restoring the ability of the mutant to utilize nitrate. Fnr1 disruption mutants showed high tolerance to chlorate and reduced ability to utilize several secondary nitrogen sources such as amino acids, hypoxanthine and uric acid, whereas growth on favourable nitrogen sources was not affected. Fnr1 disruption also abolished in vitro expression of nutrition genes , normally induced during the early phase of infection. In an infection assay on tomato seedlings, infection rate of disruption mutants was significantly delayed in comparison with the parental strain. Our results indicate that FNR1 mediates adaptation to nitrogen-poor conditions in planta through the regulation of secondary nitrogen acquisition, and as such acts as a determinant for fungal fitness during infection.     

Pod Formation and Its Geotropic Orientation in the Peanut, Arachis hypogaea L., in Relation to Light and Mechanical Stimulus

Gynophore elongation, pod formation and pod orientation in thepeanut plant (Arachis hypogaea L.) were studied in relationto the effects of light and dark conditions, mechanical stimulus,and growth substances. It was found that the proembryos controlgynophore elongation, probably by secretion of growth regulatorswhich stimulate cell division in the intercalary meristem locatedproximal to the ovules. The stimulus of pod production causesthe development of the proembryo into a mature embryo simultaneouslywith the growth of pod tissues and the cessation of gynophoreelongation. Darkness was found to be an essential factor forthe induction of pod formation. Pod formation did not occurin any of the treatments performed in the light, including theapplication of different growth substances on the ovary. A mechanicalstimulus is needed, in addition to darkness, for the normalthickening and diageotropic orientation of the pod, caused bya higher growth rate of the basal proximal side of the pod.The two ovules are always located on the upper wall of the diageotropicallyoriented pod (ventral suture). A possible mechanism which causessuch an orientation is discussed.     

Hormonal Regulation of Cormel Formation in Gladiolus Stolons Grown in vitro

The influence of four plant hormones on cormel development inGladiolus stolon tips grown in vitro was tested. Kinetin inducedcormel formation while Gibberellin A₃ inhibited it at low kinetinlevels. Abscisic acid did not affect cormel formation, but inhibitedtheir growth. Naphthalene acetic acid had no direct effect ontuberization. Anatomical observations revealed no differencesbetween cormels formed in vitro and in vivo.