Transcriptomics of Arabidopsis sperm cells at single-cell resolution

Misra, Chandra Shekhar; Santos, Mário R.; Rafael-Fernandes, Mariana; Martins, Nuno P.; Monteiro, Marta; Becker, Jörg

doi:10.1007/s00497-018-00355-4

Cited by 23 publications

(21 citation statements)

References 56 publications

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“…Upon germination the pollen tube is developed by the vegetative cell protruding out of the pollen grain and represents a specialized tubular cell that individually performs tip growth, thereby penetrating the female tissue and transporting two male gametes within its cytoplasm through the pistil to the ovule for double fertilization (Borg, Brownfield, & Twell, 2009). Despite the fact that these sperm cells have highly condensed chromatin they do not represent a totally passive cargo as formerly suggested, but it has been shown that they are transcriptionally active and required for efficient pollen tube guidance to the ovule (Borges et al, 2008; Engel, Holmes‐Davis, & McCormick, 2005; Misra et al, 2019; Santos, Bispo, & Becker, 2017; von Besser, Frank, Johnson, & Preuss, 2006). However, the extent of interaction between sperm and pollen tube is still a matter of debate, as for instance Arabidopsis gametophyte drop1/2 knockout mutants have been shown to produce a functional pollen tube without sperm cells (Zhang et al, 2017).…”

Section: Introductionmentioning

confidence: 99%

Implications of ionizing radiation on pollen performance in comparison with diverse models of polar cell growth

Stephan

2020

Plant Cell & Environment

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Research concerning the effects of ionizing radiation (IR) on plant systems is essential for numerous aspects of human society, as for instance, in terms of agriculture and plant breeding, but additionally for elucidating consequences of radioactive contamination of the ecosphere. This comprehensive survey analyses effects of x‐ and γ‐irradiation on male gametophytes comprising primarily in vitro but also in vivo data of diverse plant species. The IR‐dose range for pollen performance was compiled and 50% inhibition doses (ID50) for germination and tube growth were comparatively related to physiological characteristics of the microgametophyte. Factors influencing IR‐susceptibility of mature pollen and polarized tube growth were evaluated, such as dose‐rate, environmental conditions, or species‐related variations. In addition, all available reports suggesting bio‐positive IR‐effects particularly on pollen performance were examined. Most importantly, for the first time influences of IR specifically on diverse phylogenetic models of polar cell growth were comparatively analysed, and thus demonstrated that the gametophytic system of pollen is extremely resistant to IR, more than plant sporophytes and especially much more than comparable animal cells. Beyond that, this study develops hypotheses regarding a molecular basis for the extreme IR‐resistance of the plant microgametophyte and highlights its unique rank among organismal systems.

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Section: Introductionmentioning

confidence: 99%

Implications of ionizing radiation on pollen performance in comparison with diverse models of polar cell growth

Stephan

2020

Plant Cell & Environment

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“…The pistil was incubated for an additional 4 hours for the pollen tubes to emerge from the cut end of the style. The pollen tubes were harvested using a 25G needle and immediately frozen in liquid nitrogen and subsequently used for the RNA-seq library preparation as described in 74 .…”

Section: Methodsmentioning

confidence: 99%

“…Sterilized sporophytes were then broken using a pipette tip and diluted into 5mL molecular grade water. Spore containing solution was then distributed into 4 sterile peat pellets (Jiffy-7, Jiffy Products International) and two 9 cm Petri dishes containing KNOPS medium RNA-seq library preparation for all samples was performed as described in 74 , with the addition of mixing the PCR tubes on a Thermomixer C (Eppendorf) every 15 minutes at 200 rpm for 1 min during the RT step.…”

Section: Plant Growthmentioning

confidence: 99%

Comparative transcriptomic analysis reveals conserved transcriptional programs underpinning organogenesis and reproduction in land plants

Julca

Ferrari

Flores-Tornero

et al. 2020

Preprint

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The evolution of plant organs, including leaves, stems, roots, and flowers, mediated the explosive radiation of land plants, which shaped the biosphere and allowed the establishment of terrestrial animal life. Furthermore, the fertilization products of angiosperms, seeds serve as the basis for most of our food. The evolution of organs and immobile gametes required the coordinated acquisition of novel gene functions, the co-option of existing genes, and the development of novel regulatory programs. However, our knowledge of these events is limited, as no large-scale analyses of genomic and transcriptomic data have been performed for land plants. To remedy this, we have generated gene expression atlases for various organs and gametes of 10 plant species comprising bryophytes, vascular plants, gymnosperms, and flowering plants. Comparative analysis of the atlases identified hundreds of organ- and gamete-specific gene families and revealed that most of the specific transcriptomes are significantly conserved. Interestingly, the appearance of organ-specific gene families does not coincide with the corresponding organ's appearance, suggesting that co-option of existing genes is the main mechanism for evolving new organs. In contrast to female gametes, male gametes showed a high number and conservation of specific genes, suggesting that male reproduction is highly specialized. The expression atlas capturing pollen development revealed numerous transcription factors and kinases essential for pollen biogenesis and function. To provide easy access to the expression atlases and these comparative analyses, we provide an online database, www.evorepro.plant.tools, that allows the exploration of expression profiles, organ-specific genes, phylogenetic trees, co-expression networks, and others.

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“…Contaminating somatic tissue may not be problematic in some applications, but it can be in others, such as the estimation of 2n gamete frequencies [10,11] or tissue-specific gene expression studies [12]. In such cases, careful flower dissection may be necessary, or flow cytometry (FCM) sorting may be used to further purify samples (Section 2.1).…”

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confidence: 99%

Flow cytometric analysis of pollen and spores: An overview of applications and methodology

et al. 2021

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Pollen grains are the male gametophytes in a seed-plant life cycle. Their small, particulate nature and crucial role in plant reproduction have made them an attractive object of study using flow cytometry (FCM), with a wide range of applications existing in the literature. While methodological considerations for many of these overlap with those for other tissue types (e.g., general considerations for the measurement of nuclear DNA content), the relative complexity of pollen compared to single cells presents some unique challenges. We consider these here in the context of both the identification and isolation of pollen and its subunits, and the types of research applications. While the discussion here mostly concerns pollen, the general principles described here can be extended to apply to spores in ferns, lycophytes, and bryophytes. In addition to recommendations provided in more general studies, some recurring and notable issues related specifically to pollen and spores are highlighted.

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Transcriptomics of Arabidopsis sperm cells at single-cell resolution

Cited by 23 publications

References 56 publications

Implications of ionizing radiation on pollen performance in comparison with diverse models of polar cell growth

Implications of ionizing radiation on pollen performance in comparison with diverse models of polar cell growth

Comparative transcriptomic analysis reveals conserved transcriptional programs underpinning organogenesis and reproduction in land plants

Flow cytometric analysis of pollen and spores: An overview of applications and methodology

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