Characterization of self-incompatibility in Campanula rapunculoides (Campanulaceae) through genetic analyses and microscopy

Botany ◽  
2008 ◽  
Vol 86 (1) ◽  
pp. 1-13 ◽  
Author(s):  
S. V. Good-Avila ◽  
D. Majumder ◽  
H. Amos ◽  
A. G. Stephenson
Keyword(s):  
Genetic Basis ◽  
Male Gametophyte ◽  
Single Gene ◽  
Pollen Grains ◽  
Diallel Analysis ◽  
Pollen Tubes ◽  
Self Incompatibility ◽  
The Family

In this paper, we seek to identify the genetic basis of self-incompatibility (SI) in Campanula rapunculoides L. through diallel analysis of full siblings; to characterize the growth of pollen tubes in vivo after incompatible and compatible pollination; and to determine whether the SI system is based on pistil S-RNases. Pollinations were performed among individuals from five diallel crosses and scored for both fruit set and pollen-tube growth to determine the genetic basis of SI. On a subset of these individuals with known cross-(in)compatibility relationships, additional crosses were performed and pistils collected 1, 3, 6, 12, and 24 h after pollination to assess both the percentage of pollen grains that had germinated on the stigma, and the number of pollen tubes that had grown 20%, 40% 60%, 80%, and 100% of the distance down the pistil over five time intervals. Finally, total pistillate proteins were extracted and subjected to isoelectric focusing and RNase activity staining to find evidence of a highly basic S-RNases associated with SI in the Solanaceae. We found evidence that the SI system was based on the haplotype of the male gametophyte, and was not sporophytic. Protein analyses showed that SI was not based on a pistillate S-RNase. The existence of modifiers of SI and possible polyploidy at the S-locus complicated the expression of SI in this species, and single-gene inheritance could not be determined. This represents the first published characterization of incompatibility in the family Campanulaceae.

scholarly journals Isolation of living sperm cells and in vitro fusion of Torenia fournieri gametes

2006 ◽  
Vol 12 (4) ◽  
Author(s):  
P. Vági ◽  
K. Imre ◽  
Z. Kristóf
Keyword(s):  
Male Gametophyte ◽  
Embryo Sac ◽  
Pollen Tubes ◽  
Sperm Cells ◽  
Torenia Fournieri ◽  
Isolation Medium ◽  
Vitro Fertilization ◽  
Crucial Part

In contrast to most angiosperms, Torenia contains a naked embryo sac and therefore has been considered since many years as an exciting model plant to study the double fertilization process of flowering seed plants. It is thus not surprising that the isolation of protoplasts from the female gametophyte has been reported already 20 years ago by Mol, the isolation of megaspores and megagametophytes has been published by the authors of this manuscript in 1996 and in 1999. The isolation of the male gametophyte and of sperm cells was published by the authors in 2004. The isolation of viable Torenia sperm cells is a crucial part of the elaboration of an in vitro fertilization system. Torenia sperm cells were isolated from in vivo — in vitro cultured pollen tubes. In this system pollen tubes first grow inside a cut style then follow their elongation in a solid isolation medium. The medium contained agarose in order to detain pollen tube contents. Released sperm cells and enzymatically isolated egg cells were collected and handled using glass micropipettes and transmitted to an electrofusion apparatus or polyethylene glycol containing media for fusion probes.

scholarly journals Gene Models, Expression Repertoire, and Immune Response of Plasmodium vivax Reticulocyte Binding Proteins

2015 ◽  
Vol 84 (3) ◽  
pp. 677-685 ◽  
Author(s):  
Jenni Hietanen ◽  
Anongruk Chim-ong ◽  
Thanprakorn Chiramanewong ◽  
Jakub Gruszczyk ◽  
Wanlapa Roobsoong ◽  
...  
Keyword(s):  
Plasmodium Vivax ◽  
Significant Negative Correlation ◽  
Protein Coding ◽  
Content Type ◽  
Immune Pressure ◽  
The Family ◽  
Genes Encoding ◽  
Gene Models

Members of thePlasmodium vivaxreticulocyte binding protein (PvRBP) family are believed to mediate specific invasion of reticulocytes byP. vivax. In this study, we performed molecular characterization of genes encoding members of this protein family. Through cDNA sequencing, we constructed full-length gene models and verified genes that are protein coding and those that are pseudogenes. We also used quantitative PCR to measure theirin vivotranscript abundances in clinicalP. vivaxisolates. Like genes encoding related invasion ligands ofP. falciparum,Pvrbpexpression levels vary broadly across different parasite isolates. Through antibody measurements, we found that host immune pressure may be the driving force behind the distinctly high diversity of one of the family members, PvRBP2c. Mild yet significant negative correlation was found between parasitemia and the PvRBP2b antibody level, suggesting that antibodies to the protein may interfere with invasion.

scholarly journals Hepatoprotective Activity of Leptadenia hastata (Asclepiadaceae) on Acetaminophen-Induced Toxicity in Mice: In Vivo Study and Characterization of Bioactive Compounds through Molecular Docking Approaches

2020 ◽  
Vol 2020 ◽  
pp. 1-15 ◽  
Author(s):  
Borris R. T. Galani ◽  
Brice A. Owona ◽  
Dieudonné P. D. Chuisseu ◽  
Esaïe Machewere ◽  
Claude B. N. Ngantchouko ◽  
...  
Keyword(s):  
Molecular Docking ◽  
In Silico ◽  
Hepatoprotective Activity ◽  
Negative Control ◽  
Binding Affinities ◽  
Necrosis Factor Alpha ◽  
The Family ◽  
Factor Alpha

Background and Objectives. Leptadenia hastata is a liana from the family of Asclepiadaceae used in tropical Africa to treat diabetes mellitus. In this study, we investigated its hepatoprotective mechanisms on acetaminophen- (APAP-) induced toxicity through in vivo and in silico approaches. Materials and Methods. Various aqueous extracts were prepared from this plant and preadministered per os to albino mice 3 h before APAP administration, once daily for one week. Animals from the normal group were given only distilled water while those from negative control received only APAP 250 mg/kg. After treatment, mice were sacrificed, the liver was collected for histopathology analysis, and different biochemical markers (alanine aminotransferase (ALT), superoxide dismutase (SOD), catalase (CAT), glutathione (GSH), malondialdehyde (MDA), and tumor necrosis factor-alpha (TNFα)) were measured. The content of the active extract was analyzed by HPLC/UV. Molecular docking was conducted using iGEMDOCK software, and the drug-likeness and pharmacokinetic profiles were evaluated using Swiss ADME. Results. APAP administration significantly increased (p<0.001) ALT in liver homogenates when compared to normal controls whereas the stem decoction at 250 mg/kg significantly (p<0.001) reduced this activity to a normal value comparable to silymarin 50 mg/kg which is better than leaf and root extracts. Moreover, the stem decoction also significantly reduced the MDA levels (p<0.05) and increased those of GSH, SOD, and CAT (p<0.001) at doses of 250 and 500 mg/kg compared to the negative control. A significant (p<0.001) decrease of TNFα levels and leukocyte infiltration was observed following treatment with this extract. The HPLC/UV analysis of the decoction revealed the presence of dihydroxycoumarin, quinine, and scopoletin with the following retention times: 2.6, 5.1, and 7.01 min, respectively. In silico studies showed that quinine and dihydroxycoumarin had great potentials to be orally administered drugs and possessed strong binding affinities with TNFα, TNF receptor, cyclooxygenase-2, iNOS, cytochrome P450 2E1, and GSH reductase. Conclusion. Based on these results, L. hastata could be considered a source of promising hepatoprotective compounds with antioxidant and anti-inflammatory properties.

Isolation of male and female gametes, zygotes and proembryos of leek (Allium tuberosum Roxb)

Zygote ◽  
2020 ◽  
Vol 28 (4) ◽  
pp. 278-285
Author(s):  
Yi Hua Lin ◽  
Mei Zhen Lin ◽  
Yu Qing Chen ◽  
Hui Qiao Tian
Keyword(s):  
Higher Plants ◽  
Osmotic Shock ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Development Stage ◽  
Male And Female ◽  
Allium Tuberosum ◽  
Biological Methods ◽  
Generative Cells

SummaryThe isolation of male and female gametes is an effective method to study the fertilization mechanisms of higher plants. An osmotic shock method was used to rupture pollen grains of Allium tuberosum Roxb and release the pollen contents, including generative cells, which were mass collected. The pollinated styles were cut following 3 h of in vivo growth, and cultured in medium for 6–8 h, during which time pollen tubes grew out of the cut end of the style. After pollen tubes were transferred into a solution containing 6% mannitol, tubes burst and released pairs of sperm cells. Ovules of A. tuberosum were incubated in an enzyme solution for 30 min, and then dissected to remove the integuments. Following transfer to a dissecting solution free of enzymes, each nucellus was cut in the middle, and squeezed gently on the micropylar end, resulting in the liberation of the egg, zygote and proembryo from ovules at selected stages. These cells can be used to explore fertilization and embryonic development using molecular biological methods for each cell type and development stage.

scholarly journals Investigating mechanisms involved in the self–incompatibility response in Papaver rhoeas

2003 ◽  
Vol 358 (1434) ◽  
pp. 1033-1036 ◽  
Author(s):  
Steve Thomas ◽  
Kim Osman ◽  
Barend H. J. de Graaf ◽  
Galina Shevchenko ◽  
Mike Wheeler ◽  
...  
Keyword(s):  
Actin Cytoskeleton ◽  
Nuclear Dna ◽  
Male Gametophyte ◽  
Mitogen Activated Protein Kinase ◽  
Self Incompatibility ◽  
Papaver Rhoeas ◽  
Incompatible Pollen ◽  
Mitogen Activated Protein ◽  
Signalling Cascade

Sexual reproduction in flowering plants is controlled by recognition mechanisms involving the male gametophyte (the pollen) and the female sporophyte (the pistil). Self–incompatibility (SI) involves the recognition and rejection of self– or incompatible pollen by the pistil. In Papaver rhoeas , SI uses a Ca 2+ –based signalling cascade triggered by the S –protein, which is encoded by the stigmatic component of the S –locus. This results in the rapid inhibition of incompatible pollen tube growth. We have identified several targets of the SI signalling cascade, including protein kinases, the actin cytoskeleton and nuclear DNA. Here, we summarize progress made on currently funded projects in our laboratory investigating some of the components targeted by SI, comprising (i) the characterization of a pollen phosphoprotein (p26) that is rapidly phosphorylated upon an incompatible SI response; (ii) the identification and characterization of a pollen mitogen–activated protein kinase (p56), which exhibits enhanced activation during SI; (iii) characterizing components involved in the reorganization and depolymerization of the actin cytoskeleton during the SI response; and (iv) investigating whether the SI response involves a programmed cell death signalling cascade.

scholarly journals Embryological studies of reciprocal crosses between Vicia faba and Vicia narbonensis

2014 ◽  
Vol 67 (1) ◽  
pp. 37-43 ◽  
Author(s):  
Maciej Zenkteler ◽  
Mechtild Tegeder ◽  
Otto Schieder
Keyword(s):  
Vicia Faba ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Flower Buds ◽  
Reciprocal Crosses ◽  
Globular Embryos ◽  
Vicia Narbonensis ◽  
Whole Number

A study was undertaken to asses the reciprocal crossability between <em>Vicia faba</em> and <em>Vicia narbonensis</em>. Flower buds or only ovaries of several varietes and genotypes were cross-pollinated in vivo (green house and field) and in vitro. Only few pollen tubes passed the style and entered into the ovary. On the whole number of 5320 cross pollinated in vivo and in vitro flowers and ovaries of <em>Vicia narbonensis</em> only 78 globular hybrid embryos were observed. After cross pollination in vivo of 3860 flower buds and ovaries of <em>Vicia faba</em> globular embryos developed in 124 ovules. The highest number of globular embryos were obtained when the <em>Vicia faba</em> line 1/33 was pollinated with <em>Vicia narbonensis</em> lines P3, P5, 150, SE.Embryogenesis proceeded till the 6-10 day after pollination, however, karyological disturbances in the cells of embryos and endosperm were often noticed at earlier stages. In vitro pollen grains of <em>Vicia faba</em> germinated on stigmas and ovaries of <em>Vicia narbonensis</em>, a significant increase in the growth of pollen tubes was noticed after ovary pollination. The technique of in vitro pollination was not suitable for <em>Vicia faba</em> as the inoculated explants died shortly after transferring onto the medium. The results indicate that finding a more suitable genotype for crossing may give a chance to obtain higher number of embryos (example line 1/33) - thus sufficient number for culturing them on media.

scholarly journals 455 PB 326 PISTIL INFLUENCE ON GROWTH OF POLLEN TUBES OF P. X DOMESTICUM

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 496e-496
Author(s):  
Mary Stuart ◽  
Pablo Jourdan
Keyword(s):  
Male Gametophyte ◽  
Genotypic Variation ◽  
Pollen Grains ◽  
Pollen Tubes ◽  
Female Fertility ◽  
Low Fertility ◽  
High Fertility ◽  
Poor Seed ◽  
Germinated Pollen ◽  
Germination And Growth

The regal pelargonium (P. x domesticum) is generally characterized by low fertility and poor seed set. In studys designed to assess factors that contribute to low fecundity in this crop we have examined genotype interactions among various cultivars and have identified lines that differ in degree of male and female fertility. The objective of this study was to examine genotypic variation, other than self-incompatibility, of P. x domesticum pistils in supporting the development of the male gametophyte. Variation in pollen germination and growth was assessed after crossing either a male of high fertility or a mate of poor fertility to nine different selections of varying female fertility. Styles were harvested 2 hours after pollination and examined using fluorescence microscopy to determine the number of germinated pollen grains on the stigma and the number of pollen tubes growing down the style. Female selections displayed large differences in their ability to support pollen tubes. Styles from different females pollinated with the same male varied in average number of pollen tubes from 30 to 2.

scholarly journals A primary study on genes with selected mutations by in vitro passage of Chlamydia muridarum strains

2019 ◽  
Vol 77 (3) ◽  
Author(s):  
Zhou Zhou ◽  
Na Liu ◽  
Yingzi Wang ◽  
Arthur Wirekoh Emmanuel ◽  
Xiaoxing You ◽  
...  
Keyword(s):  
Single Gene ◽  
Point Mutations ◽  
Primary Study ◽  
Chlamydia Muridarum ◽  
In Vivo Experiments ◽  
High Incidence ◽  
Identification And Characterization

ABSTRACTObjectiveThis study is to investigate the functions of newly discovered genes in Chlamydia muridarum (C. muridarum) strains with single gene differences.MethodsUsing whole genome sequencing and plaque formation assays, C. muridarum parental and passaging strains were established, and the isogenic clones expressing certain genotypes were isolated. Strains with single gene differences were obtained. Based on prediction, the valuable strains with single gene differences of tc0412, tc0668 or tc0237 were subjected to the in vitro and in vivo experiments for biological characterization and virulence analysis.ResultsInsertional -472840T mutation of the tc0412 gene (T28T/B3 type) matching with the nonmutant tc0668 gene and tc0237 gene with point mutations G797659T (Q117E) might slow the growth of Chlamydia due to the lack of a plasmid. The nonmutant tc0668 in the strain might induce a high incidence of hydrosalpinx in mice, while tc0668 with a G797659T point mutation was significantly attenuated. Compared with the nonmutant tc0237, the strains containing mutant tc0237 were characterized by reduced centrifugation dependence during infection.ConclusionThe identification and characterization of these genes might contribute to the comprehensive understanding of the pathogenic mechanism of Chlamydia.

scholarly journals Maize ROP2 GTPase Provides a Competitive Advantage to the Male Gametophyte

Genetics ◽  
2003 ◽  
Vol 165 (4) ◽  
pp. 2137-2151 ◽  
Author(s):  
K M Arthur ◽  
Z Vejlupkova ◽  
R B Meeley ◽  
J E Fowler
Keyword(s):  
Male Gametophyte ◽  
Transmission Rate ◽  
Pollen Grains ◽  
Duplicate Gene ◽  
Wild Type ◽  
Allelic Series ◽  
Plant Signal Transduction ◽  
Relative Transmission ◽  
Mutant Pollen

Abstract Rop GTPases have been implicated in the regulation of plant signal transduction and cell morphogenesis. To explore ROP2 function in maize, we isolated five Mutator transposon insertions (rop2::Mu alleles). Transmission frequency through the male gametophyte, but not the female, was lower than expected in three of the rop2::Mu mutants. These three alleles formed an allelic series on the basis of the relative transmission rate of each when crossed as trans-heterozygotes. A dramatic reduction in the level of ROP2-mRNA in pollen was associated with the three alleles causing a transmission defect, whereas a rop2::Mu allele that did not result in a defect had wild-type transcript levels, thus confirming that mutation of rop2 causes the mutant phenotype. These data strongly support a role for rop2 in male gametophyte function, perhaps surprisingly, given the expression in pollen of the nearly identical duplicate gene rop9. However, the transmission defect was apparent only when a rop2::Mu heterozygote was used as the pollen donor or when a mixture of wild-type and homozygous mutant pollen was used. Thus, mutant pollen is at a competitive disadvantage compared to wild-type pollen, although mutant pollen grains lacked an obvious cellular defect. Our data demonstrate the importance in vivo of a specific Rop, rop2, in the male gametophyte.

scholarly journals Identification and Characterization of a Polymorphic Receptor Kinase Gene Linked to the Self-Incompatibility Locus of Arabidopsis lyrata

Genetics ◽  
2001 ◽  
Vol 158 (1) ◽  
pp. 387-399 ◽  
Author(s):  
Mikkel H Schierup ◽  
Barbara K Mable ◽  
Philip Awadalla ◽  
Deborah Charlesworth
Keyword(s):  
Brassica Species ◽  
Self Incompatibility ◽  
Kinase Gene ◽  
Arabidopsis Lyrata ◽  
The Family ◽  
S Alleles ◽  
Sib Families ◽  
Identification And Characterization ◽  
Incompatibility Locus

Abstract We study the segregation of variants of a putative self-incompatibility gene in Arabidopsis lyrata. This gene encodes a sequence that is homologous to the protein encoded by the SRK gene involved in self-incompatibility in Brassica species. We show by diallel pollinations of plants in several full-sib families that seven different sequences of the gene in A. lyrata are linked to different S-alleles, and segregation analysis in further sibships shows that four other sequences behave as allelic to these. The family data on incompatibility provide evidence for dominance classes among the S-alleles, as expected for a sporophytic SI system. We observe no division into pollen-dominant and pollen-recessive classes of alleles as has been found in Brassica, but our alleles fall into at least three dominance classes in both pollen and stigma expression. The diversity among sequences of the A. lyrata putative S-alleles is greater than among the published Brassica SRK sequences, and, unlike Brassica, the alleles do not cluster into groups with similar dominance.

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