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Gametophytes

Gametophytes are the haploid, multicellular stage in the life cycle of plants, ferns, and some other organisms.
They arise from the germination of spores and are responsible for sexual reproduction, producing gametes that fuse to form the diploid sporophyte.
Gametophytes vary in size and complexity, from the tiny, free-living plants of mosses and liverworts to the large, independent structures of some ferns.
Understanding the biology and development of gametophtes is crucial for research in plant evolutionary biology, reproductive biology, and ecologey.
This MeSH term provides a concise overview of this important plant life stage.

Most cited protocols related to «Gametophytes»

Establishing Arabidopsis Single Mutant Phenotypes

To establish the primary dataset of Arabidopsis (Arabidopsis thaliana) genes with a single mutant phenotype, we started with a published list of 620 Arabidopsis genes included in our sequence-based map of genes with mutant phenotypes (Meinke et al., 2003 (link)), removed problematic loci with questionable genotype-to-phenotype associations, eliminated suppressors and genes with a dominant gain-of-function mutant phenotype but no apparent loss-of-function phenotype, and further curated the phenotype and gene function information. Several classical genetic loci with well-characterized dominant phenotypes (e.g. GAI, ETR1, ABI1) were retained in the dataset because they are also associated with a distinctive loss-of-function phenotype (Peng et al., 1997 (link); Cancel and Larsen, 2002 (link); Ludwików et al., 2009 (link)). We then requested from Eva Huala at TAIR a list of genes that appeared to be associated with phenotype information in the TAIR database. Each locus on the list was evaluated. Most entries yielded useful information, but many candidate genes were eliminated because no suitable phenotype information was found or because the locus did not code for a protein. To complement these efforts, we initiated extensive PubMed searches of the scientific literature, using a combination of the following keywords: Arabidopsis, mutant(s), mutation(s), knockout, and null. Several thousand articles were retrieved and analyzed to obtain the information presented in Supplemental Table S2. Information on genes with multiple mutant phenotypes was also retrieved with this approach. In order to proceed with further analysis of these datasets, no literature searches were performed for publications added to the PubMed database after December 31, 2010.
We then updated the information on essential genes based on the eighth release of the SeedGenes database (www.seedgenes.org). Additional updates were obtained from a recent publication on embryo and gametophyte essentials of Arabidopsis (Muralla et al., 2011 (link)). We classified essential genes as being required for early development or survival. A locus was considered to be essential when knockout heterozygotes segregated for defective embryos or gametophytes, regardless of whether the resulting homozygotes remained viable to the seedling stage or beyond. To be consistent with the prioritized classification system established here, EMB loci with defects in gametophyte function were assigned to the gametophyte class instead of the seed/embryo class, regardless of whether the locus was classified elsewhere as being required for seed development, because that is when the mutant phenotype was first detected. The criteria used to differentiate between the GAM, GEM, EMG, and EMB subsets of essential genes are detailed elsewhere (Muralla et al., 2011 (link)). Mutants with defective gametes that produced viable homozygotes were typically assigned to the MGD subset. The criteria used to make other phenotype subset assignments, listed in Supplemental Table S1, can be gauged by accessing the second tabbed spreadsheet in Supplemental Table S2, sorting for the subset of interest, and evaluating the diversity of phenotypes represented.

Lloyd J, & Meinke D. (2012). A Comprehensive Dataset of Genes with a Loss-of-Function Mutant Phenotype in Arabidopsis. Plant Physiology, 158(3), 1115-1129.

Publication 2012

ABI1 protein, human Arabidopsis Arabidopsis thalianas Chromosome Mapping Embryo Gametes Gametophytes Genes Genes, Dominant Genes, Essential Genes, vif Genetic Loci Heterozygote Homozygote Mutation Operator, Genetic Phenotype Staphylococcal Protein A

Comparison of Sporophyte and Gametophyte Transcriptomes

Sporophytic data from public baseline GeneChip experiments used for comparison with the pollen transcriptome were downloaded from the NASC website [41 ,64 (link)]. The list of dataset codes was as follows: COT (three replicates), Cornah_A4-cornah-wsx_SLD_REP1-3; LEF (three replicates), A4-LLOYD-CON_REP1-3; PET (three replicates), Millenaar_A1-MILL-AIR-REP1-3; STM (two replicates), Turner_A-7-Turne-WT-Base1-2_SLD; ROT (two replicates), Sophie_A1-Fille-WT-nodex_SLD, Sophie_A5-Fille-WT-nodex_SLD; RHR (two replicates), Jones_A1-jones-WT1, SLD, Jones_A1-jones-WT2_SLD; SUS (three replicates), A1-WILLA-CON-REP1-3.
All gametophytic and sporophytic datasets were normalized using freely available dChip 1.3 software [65 ]. The reliability and reproducibility of analyses was ensured by the use of duplicates or triplicates in each experiment, the normalization of all 26 arrays to the median probe intensity level and the use of normalized CEL intensities of all arrays for the calculation of model-based gene-expression values based on the Perfect Match-only model [66 (link),67 (link)]. A given gene was scored as 'expressed' when it gave a reliable expression signal in all replicates. Expression signal value '0' means that the detection call value was not 'present' in all replicates provided. All raw and dChip-normalized gametophytic datasets are available at the Institute of Experimental Botany AS CR website [68 ]. Although a RT-PCR validation of microarray data was not performed specifically for the purpose of this publication, our confidence in the quality of the data presented is based on our previously published RT-PCR validation of the expression of 70 genes [21 (link),35 (link),41 ].
Microsoft Excel was used to manage and filter the microarray data. For annotation of genes present on the ATH1 Array, the Arabidopsis Genome Annotation Release 3.0 published by The Institute for Genomic Research [52 ] was used. Genes were sorted into functional categories created according to data mined from the Munich Information Center for Protein Sequences Arabidopsis thaliana Database [69 ], Kyoto Encyclopedia of Genes and Genomes [70 ] and TAIR [46 ]. Hierarchical clustering of expressed genes was performed using expression-profile data clustering and analysis software EPCLUST [71 ], with correlation measure based distance and average linkage clustering methods.

Honys D, & Twell D. (2004). Transcriptome analysis of haploid male gametophyte development in Arabidopsis. Genome Biology, 5(11), R85.

Publication 2004

Arabidopsis Arabidopsis thalianas Gametophytes Gene Annotation Gene Chips Gene Expression Genes Genome Microarray Analysis Pollen Reverse Transcriptase Polymerase Chain Reaction Transcriptome

Polyploidy Dynamics in Potentilla puberula

Potentilla puberula Krašan (= Potentilla pusilla Host: Soják, 2010; Figure 1) constitutes a suitable model to study the consequences of reproductive interference for co‐existence of reproductively differentiated cytotypes. The species comprises tetraploids being almost exclusively sexual and self‐incompatible and penta‐ to octoploids which are preferentially apomictic (Dobeš, Milosevic, et al., 2013; Prohaska, 2013). A screen of 269 populations along a latitudinal transect through the Eastern European Alps revealed about every second population to be cytologically mixed (i.e., inhabited by 2–5 cytotypes in various combinations). Nevertheless, the presence of tetraploids in a population was negatively related to the presence of penta‐ to octoploids and vice versa. In contrast, the occurrences of penta‐ to octoploids were hardly related to each other (Hülber, Scheffknecht, & Dobeš, 2013).
Potentilla puberula is assumed to be of allopolyploid origin with tetraploids (Soják, 2010) constituting the lowest ploidy level (Dobeš, 1999; Dobeš, Milosevic, et al., 2013). Functionality of the gametophytic SI system suggested that the tetraploids are functional diploids (Dobeš, Milosevic, et al., 2013). The cytotypes, however, are genetically barely differentiated (Paule, Scherbantin, & Dobeš, 2012) suggesting an intraspecific origin. Two polar nuclei contribute to the endosperm of both sexually and apomictically derived seed and either one or two of the sperm in the latter (Dobeš, Milosevic, et al., 2013). Pollen is mostly meiotically reduced in P. puberula irrespective of reproductive mode and ploidy of individuals (Christoph Dobeš and Christina Sykora unpublished research). Hence, the following parental genomic ratios in the endosperm constitute the normal condition in this system: 2m:1p in the tetraploid sexual cytotype with m and p representing two chromosome sets (half the number of chromosome sets present in the maternal genome), and 4m:1p or 4m:2p in the apomictic cytotypes with m and p representing half the number of their five, six, seven, and eight chromosome sets, respectively.

Dobeš C., Scheffknecht S., Fenko Y., Prohaska D., Sykora C, & Hülber K. (2017). Asymmetric reproductive interference: The consequences of cross‐pollination on reproductive success in sexual–apomictic populations of Potentilla puberula (Rosaceae). Ecology and Evolution, 8(1), 365-381.

Publication 2017

Apomixis Autoimmune Lymphoproliferative Syndrome Cell Nucleus Chromosomes Diploidy Eastern European People Endosperm Gametophytes Genome natural heparin pentasaccharide Parent Pollen Potentilla Reproduction Sperm Tetraploidy

Transcriptome Analysis of Syntrichia caninervis Dehydration-Rehydration

S. caninervis gametophytes were collected from the Gurbantunggut Desert of Xinjiang Uygur Autonomous Region of China (44° 32′ 30″ N, 88° 6′ 42″ E) and harvested and stored as described previously [14 ]. Since 2003, this sand dune has been identified as a permanent research site. In this study, patches of S. caninervis inhabiting the biological soil crusts were collected in petri dishes and stored in an air-dried state for at least 1 week at room temperature. All samples used in the experiment were collected from the same site within a 10 m² plot. Voucher specimens are maintained in the Department of Plant Biology, Southern Illinois University (Carbondale, IL). To obtain a comprehensive transcriptome assembly of S. caninervis transcripts during the dehydration-rehydration process, an equal mixture of total RNAs isolated from various dehydration and rehydration time points were used to construct the sequencing library. Dried gametophyte tissue samples were placed on filter paper in petri dishes and rehydrated using purified water for 24 hours. Gametophores were harvested after 24 h of rehydration. Gametophores were subsequently allowed to dry on an open bench (ca 25°C, RH =25%) [74 (link)] and samples (i.e. 100 mg FW) were harvested at 0.5, 1, 1.5, 2, 4, 6, 8, 10, 12 and 24 h.
Total RNAs isolated from all samples were quality and purity assessed and pooled together for RNA-Seq [75 (link)]. Total RNAs were extracted from S. caninervis gametophyte tissue samples using Trizol Reagent (Invitrogen, USA). The resulting samples were treated with DNase I to remove any genomic DNAs. RNAs were quantified using an Agilent 2100 Bioanalyzer and checked for RNA integrity using denaturing agarose gel electrophoresis. The cDNA library was created and sequenced according to the manufacturer’s instructions (Illumina) and sequencing was performed at Beijing Genome Institute (BGI) in Shenzhen, China. Briefly, beads with Oligo(dT) were used to isolate poly(A) + mRNA after total RNA was obtained. Fragmentation buffer was added for interrupting mRNA into short fragments. First-strand cDNA was synthesized using these short fragments as templates, along with reverse transcriptase and random hexamer primer. And the second-strand cDNA was synthesized using buffer, dNTPs, RNaseH and DNA polymerase I. The resulting double stranded cDNA was then subjected to end-repair using T4 DNA polymerase, DNA polymerase I Klenow fragment, and T4 polynucleotide kinase, and ligated to adapters using T4 DNA ligase. Short fragments were purified with QIAquick PCR purification kit and eluted with EB buffer. After agarose gel electrophoresis, the suitable fragments (200 ± 50 bp) were selected as templates for bridged PCR amplification. The Illumina cBOT was used for cluster generation following the manufacturer’s instructions, and the clustered flow cell was loaded onto the sequencing machine. cDNA library products were sequenced on an Illumina HiSeq™ 2000 system.

Gao B., Zhang D., Li X., Yang H, & Wood A.J. (2014). De novo assembly and characterization of the transcriptome in the desiccation-tolerant moss Syntrichia caninervis. BMC Research Notes, 7, 490.

Publication 2014

Biopharmaceuticals Buffers cDNA Library Cells Dehydration Deoxyribonuclease I DNA DNA, Complementary DNA-Directed DNA Polymerase DNA Library DNA Polymerase I Electrophoresis, Agar Gel Gametophytes Genome Hyperostosis, Diffuse Idiopathic Skeletal mRNA, Polyadenylated Oligonucleotide Primers Oligonucleotides Plants Polynucleotide 5'-Hydroxyl-Kinase Rehydration RNA, Messenger RNA, Polyadenylated RNA-Directed DNA Polymerase RNA-Seq T4 DNA Ligase Tissues Transcriptome trizol

Orchid Tissue Harvesting and RNA Extraction

For gametophytic and embryonic tissues, orchid flowers were hand pollinated and developing ovaries were harvested at the specified day. Only the interior tissues from developing ovaries were scooped and pooled for RNA extraction. For root samples, 2-cm tip tissue containing root apical meristems was collected. For stalk samples, 5–10 cm long stalks were collected. For protocorm-like body (PLB) samples, one-month-old tissues were collected. For protocorm samples, 20- and 30-day-old tissues were pooled and collected. The collected samples were flash frozen in liquid nitrogen and stored in a freezer at −80 °C. RNA was isolated using MaestroZol RNA Plus extraction reagent (Maestrogen, USA) according to the manufacturer’s instructions. The isolated total RNA was treated with RNase-free DNase (Qiagen, USA) followed by RNeasy mini-column purification according to the manufacturer’s instructions (Qiagen, USA).

Lin H.Y., Chen J.C., Wei M.J., Lien Y.C., Li H.H., Ko S.S., Liu Z.H, & Fang S.C. (2013). Genome-wide annotation, expression profiling, and protein interaction studies of the core cell-cycle genes in Phalaenopsis aphrodite. Plant Molecular Biology, 84(1), 203-226.

Publication 2013

Deoxyribonucleases Embryo Endoribonucleases Flowers Freezing Gametophytes Human Body Meristem Nitrogen Ovary Plant Roots Specimen Collection Stalking Tissues

Most recents protocols related to «Gametophytes»

Cultivation of Radula complanata Protonema

Sterile Radula complanata protonema were germinated from spores on 20-20-20 agar plates and cultivated to obtain mature gametophytes. Voucher specimens of environmental samples used for spore collection have been deposited at the Connell Memorial Herbarium at the University of New Brunswick, Fredericton under the accession numbers 69,083 and 69,084.

Blatt-Janmaat K., Neumann S., Schmidt F., Ziegler J., Qu Y, & Peters K. (2023). Impact of in vitro phytohormone treatments on the metabolome of the leafy liverwort Radula complanata (L.) Dumort. Metabolomics, 19(3), 17.

Publication 2023

Agar Gametophytes Spores Sterility, Reproductive

Stress-Response and Growth Hormone Effects on Gametophytes

Mature gametophytes were placed on 20-20-20 agar plates supplemented with hormones and grown under full spectrum growth lights. Stress-response hormones included methyl jasmonate (MeJA, Sigma), abscisic acid (AA, Sigma), and salicylic acid (SA, Sigma) while growth hormones included 1-napthaleneaceticacid (NAA, Sigma), and 6-benzylaminopurine (BAP, Sigma). Control samples (no methanol, no hormones) were prepared for both the stress-response and growth groups. The stress-response hormones were grown for 4 months while the growth hormone treatments were grown for 3 months. Plants were weighed and stored at -80 °C until further processing. Harvesting yielded the following samples: AA1, AA10, AA100, MeJA1, MeJA10, MeJA100, SA1, SA10, SA100, Stress Control, BAP1, BAP10, BAP100, NAA1, NAA10, NAA100, Growth Control. Statistical analyses were conducted in R and significant changes were determined by a one-way ANOVA and a Tukey HSD post-hoc test. Treatment fresh weights were normalized with Eq. (1) for data visualization.

Publication 2023

Abscisic Acid Agar benzylaminopurine Gametophytes Growth Hormone Hormones Light Methanol methyl jasmonate neuro-oncological ventral antigen 2, human Plants Salicylic Acid

Physiological Responses of S. caninervis

AWC was measured in gametophytes of S. caninervis at various time points during the D-R process. AWC curves, plotting the water content on a dry weight basis, were produced using the following formula (Rathnayake et al., 2019 (link)): AWC (g g^-1 DW) = (FW-DW)/DW; FW indicates the fresh weight measured at every time point during the D-R process, and DW indicates the weight measured after drying for 48 h at 80°C in an oven.
Fv/Fm was measured using a portable modulated fluorometer (PAM-2500; Heinz, Walz, Germany). The saturation pulse method was used to calculate Fv/Fm, which was measured in the dark after the box was covered for > 30 min. Parameter settings were based on the recommendations of Zhang et al. (2011) (link).
The activities of the physiological indicators H₂O₂, malondialdehyde (MDA), peroxidase (POD), and superoxide dismutase (SOD) were measured using detection assay kits (Nanjing Jiancheng Bioengineering Institute, Nanjing, China), according to the manufacturer’s instructions.

Yang R., Li X., Yang Q., Zhao M., Bai W., Liang Y., Liu X., Gao B, & Zhang D. (2023). Transcriptional profiling analysis providing insights into desiccation tolerance mechanisms of the desert moss Syntrichia caninervis. Frontiers in Plant Science, 14, 1127541.

Publication 2023

Biological Assay Gametophytes Malondialdehyde Peroxidase Peroxide, Hydrogen physiology Pulse Rate Superoxide Dismutase

Transcriptome Analysis of Dehydration-Rehydration Dynamics in Moss

Dry S. caninervis samples were collected from the Gurbantunggut Desert in Xinjiang, Northwest China (Fukang County, 44°32′30″N, 88°6′42″E). The collected wild moss samples were stored at 25°C under dark conditions. The gametophytes of these samples were completely desiccated and kept in a dormant state. For experiments, the dried wild gametophytes were, first, fully hydrated with ultrapure water for 24 h; then, the slow-dried method was used to keep S. caninervis samples at a relative humidity (RH) of 66.67% (-57 MPa) at 25°C, as described previously (Liang et al., 2021 (link)). Samples were collected after 0, 2, 6, and 24 h of dehydration (0 h as the control). Early dehydration occurred at 2 and 6 h (D2h and D6h), and late dehydration occurred at 24 h (D24h). The dehydrated samples were subsequently rehydrated by transferring the dehydrated gametophytes to new Petri dishes at 25°C with filter paper that was saturated with ultrapure water. Rehydrated samples were then harvested at 0.5, 2, 6, 24, and 48 h. Early rehydration was considered to be 0.5, and 2 h (R0.5h and R2h), whereas late rehydration times were 6, 24, and 48 h (R6h, R24h, and R48h, respectively). Fully rehydrated samples without dehydration (0 h) served as the reference control. All samples were frozen in liquid nitrogen immediately after harvest and stored at -80°C. The prepared samples were subjected to transcriptome sequencing and physiological analysis. Photographic records of S. caninveris phenotypes, in addition to the measurement of the absolute water content (AWC) and Fv/Fm (optimal/maximal photochemical efficiency of PSII) were performed at each treatment time point.

Publication 2023

Dehydration Freezing Gametophytes Humidity Hyperostosis, Diffuse Idiopathic Skeletal Mosses Nitrogen Phenotype physiology Rehydration Specimen Collection Strains

Moss Stress Response to Salinity

Axenic in vitro cultures of the experimental mosses, P. patens, E. hungaricus and H. heimii, were established and the full development of gametophores was achieved. Details of the culture procedure can be found in Sabovljević et al. [5 ,31 ] and Ćosić et al. [50 (link)]. Having achieved a sufficient number of developed gametophores to start the experiments, cultures of the selected mosses were grown on solid BCD medium (containing 0.2 M MgSO₄ × 7H₂O, 0.18 M KH₂PO₄, 1 M KNO₃, and 0.9 mM FeSO₄ × 7H₂O) with sucrose added (0.05 M) to achieve quick growth and morphologically well-developed gametophytes [5 ,52 (link)] prior to the experimental tests. The pH of the media was adjusted to 5.8 before autoclaving at 121 °C for 30 min. Further tests were conducted on minimal BCD medium supplemented with different NaCl concentrations before autoclaving (5, 10, 50, 100, 200, 300, and 500 mM NaCl). In experiment type I, the mosses were grown on BCD medium containing NaCl for 3 days, which simulated short-term stress. Experiment type II represented long-term stress, where the mosses were grown on BCD medium containing NaCl for 21 days. The control plants were grown on BCD salt-free medium. The culture conditions were set at 18 ± 2 °C, under a long-day photoperiod (16 h:8 h light/dark). All the plants tested here were grown under the same controlled laboratory conditions including the light quality and intensity (47 μmol m⁻²s⁻¹ irradiance). After 3 and 21 days, respectively, the plant material was collected and frozen at −70 °C until further analysis. All the chemicals were supplied by Sigma Aldrich, Germany.

Ćosić M.V., Mišić D.M., Jakovljević K.M., Giba Z.S., Sabovljević A.D., Sabovljević M.S, & Vujičić M.M. (2023). Analysis of the Qualitative and Quantitative Content of the Phenolic Compounds of Selected Moss Species under NaCl Stress. Molecules, 28(4), 1794.

Publication 2023

Axenic Culture Freezing Gametophytes Light Mosses Plants Salts Sodium Chloride Sucrose Sulfate, Magnesium

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More about "Gametophytes"

Gametophytes are the haploid, multicellular reproductive stage in the life cycle of many plants, ferns, and other organisms.
These structures arise from the germination of spores and are responsible for sexual reproduction, producing gametes that fuse to form the diploid sporophyte generation.
The gametophyte can vary greatly in size and complexity, ranging from the tiny, free-living plants of mosses and liverworts to the large, independent structures of some ferns.
Understanding the biology and development of gametophytes is crucial for research in plant evolutionary biology, reproductive biology, and ecology.
Gametophytes, also known as the 'gamete-bearing' or 'sexual' stage, play a vital role in the life cycle of plants, providing the foundation for sexual reproduction and genetic diversity.
Researchers studying gametophytes may utilize techniques such as the Spectrum Plant Total RNA Kit for RNA extraction, the HiSeq 2000 or HiSeq 2500 for high-throughput sequencing, the RNeasy Plant Mini Kit or RNeasy Mini Kit for RNA purification, and the Agilent 2100 Bioanalyzer for quality control.
Computational tools like SAS version 9.4 can also be employed for data analysis.
Proper filtration using a 0.22 μm filter may be necessary to ensure sample purity.
Exploring the fascinating world of gametophytes can lead to advancements in our understanding of plant evolutionary processes, reproductive strategies, and ecological adaptations.
By mastering the techniques and insights related to this crucial plant life stage, researchers can unlock new discoveries and push the boundaries of plant science.