Transcriptome profiling of tobacco (Nicotiana tabacum) pollen and pollen tubes

Conze, Lei Liu; Berlin, Sofia; Bail, Aude Le; Kost, Benedikt

doi:10.1186/s12864-017-3972-3

Cited by 25 publications

(17 citation statements)

References 47 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The observation that we detected especially cell wall enzymes and CRPs in pollen tube secretomes was not surprising taking into consideration that these belong also to the strongly expressed genes (e.g., Bokvaj et al 2015;Conze et al 2017;Tan et al 2018). Moreover, pollen tubes are fast growing plant cells that require a solid cell wall and that communicate intensively with the surrounding maternal tissues.…”

Section: Enrichment Of Cell Wall Enzymes and Crps In Pollen Tube Secrsupporting

confidence: 56%

Comparative analyses of angiosperm secretomes identify apoplastic pollen tube functions and novel secreted peptides

et al. 2020

View full text Add to dashboard Cite

Key message Analyses of secretomes of in vitro grown pollen tubes from Amborella, maize and tobacco identified many components of processes associated with the cell wall, signaling and metabolism as well as novel small secreted peptides. Abstract Flowering plants (angiosperms) generate pollen grains that germinate on the stigma and produce tubes to transport their sperm cells cargo deep into the maternal reproductive tissues toward the ovules for a double fertilization process. During their journey, pollen tubes secrete many proteins (secreted proteome or secretome) required, for example, for communication with the maternal reproductive tissues, to build a solid own cell wall that withstands their high turgor pressure while softening simultaneously maternal cell wall tissue. The composition and species specificity or family specificity of the pollen tube secretome is poorly understood. Here, we provide a suitable method to obtain the pollen tube secretome from in vitro grown pollen tubes of the basal angiosperm Amborella trichopoda (Amborella) and the Poaceae model maize. The previously published secretome of tobacco pollen tubes was used as an example of eudicotyledonous plants in this comparative study. The secretome of the three species is each strongly different compared to the respective protein composition of pollen grains and tubes. In Amborella and maize, about 40% proteins are secreted by the conventional “classic” pathway and 30% by unconventional pathways. The latter pathway is expanded in tobacco. Proteins enriched in the secretome are especially involved in functions associated with the cell wall, cell surface, energy and lipid metabolism, proteolysis and redox processes. Expansins, pectin methylesterase inhibitors and RALFs are enriched in maize, while tobacco secretes many proteins involved, for example, in proteolysis and signaling. While the majority of proteins detected in the secretome occur also in pollen grains and pollen tubes, and correlate in the number of mapped peptides with relative gene expression levels, some novel secreted small proteins were identified. Moreover, the identification of secreted proteins containing pro-peptides indicates that these are processed in the apoplast. In conclusion, we provide a proteome resource from three distinct angiosperm clades that can be utilized among others to study the localization, abundance and processing of known secreted proteins and help to identify novel pollen tube secreted proteins for functional studies.

show abstract

Section: Enrichment Of Cell Wall Enzymes and Crps In Pollen Tube Secrsupporting

confidence: 56%

Comparative analyses of angiosperm secretomes identify apoplastic pollen tube functions and novel secreted peptides

et al. 2020

View full text Add to dashboard Cite

show abstract

“…Analysis of the EXO70.1 paralogues expression in male gametophyte and sporophytic tissues was performed using the CoNekT (Proost and Mutwil, 2018) and GEO (Barrett et al, 2013) tools. Additional anther/pollen RNA-seq expression data for sorghum, Arabidopsis, poplar and tobacco were obtained from (Davidson et al, 2012;Loraine et al, 2013;Zhao et al, 2016;Conze et al, 2017), respectively.…”

Section: Phylogenetic and Expression Analysismentioning

confidence: 99%

EXO70A2 is critical for the exocyst complex function in Arabidopsis pollen

Marković

Cvrčková

Potocký

et al. 2019

Preprint

View full text Add to dashboard Cite

Pollen development, pollen grain germination and pollen tube elongation are crucial biological processes in angiosperm plants that need precise regulation to deliver sperm cells to fertilize ovules. Pollen grains undergo two major developmental switches: dehydration characterized by metabolic quiescent state, and rehydration upon pollination that leads to extraordinary metabolic and membrane trafficking activity, resulting in germination and rapid tip growth of pollen tubes. To sustain these processes, many plant housekeeping genes evolved their pollen-specific paralogs. Highly polarized secretion at a growing pollen tube tip requires the exocyst tethering complex responsible for specific targeting of secretory vesicles to the plasma membrane. Here, we describe that EXO70A2 (At5g52340) is the main exocyst EXO70 isoform in Arabidopsis pollen, which governs the conventional secretory function of the exocyst, analogically to EXO70A1 (At5g03540) in the sporophyte. Our analysis of a CRISPR-generated exo70a2 mutant revealed that EXO70A2 is essential for efficient pollen maturation, pollen grain germination and pollen tube growth. GFP:EXO70A2 was localized similarly to other exocyst subunits to the apical domain in growing pollen tube tips characterized by intensive exocytosis. Moreover, EXO70A2 could substitute for the EXO70A1 function in the sporophyte, indicating functional redundancy of these two closely related isoforms. Phylogenetic analysis revealed that the ancient duplication of EXO70A to two (or more) paralogs, one of which is highly expressed in pollen, occurred independently in monocots and dicots. In summary, EXO70A2 is a crucial component of the exocyst complex in the Arabidopsis pollen required for efficient plant sexual reproduction. KEY WORDSexocyst, EXO70A2, EXO70A1, pollen development, pollen tube, tip growth, polarized secretion ABBREVIATIONS CDS -coding sequence GFP -green fluorescent protein NBT -nitroblue tetrazolium PM -plasma membrane PME -pectin methylesterases ROS -reactive oxygen species WT -wild type

show abstract

“…Pollen development, maintenance, vesicle traffic, signaling, pollen tube activation, cytoskeletal dynamic, directional growth, stigma cell wall loosening, calcium gradient, ubiquitination signal, auxin, kinases, allergens, defense response, RNases, and S allele recognition genes were all detected. Likewise, related genes have been identified in maize, apricot, petunias, tomato, tea, tobacco, brassica, pear, Arabidopsis thaliana, almonds, and orchids pollination transcriptome analyses [19][20][21][22][23]25,29,38,[41][42][43][44] .…”

Section: Discussionmentioning

confidence: 99%

“…used to understand gamete development, specific pollen gene expression, pollen tube dynamics, pollen-stigma interactions, and candidate genes for self-incompatibility 22,[24][25][26] . While gene expression profiles are able to distinguish between self and outcross pollination, the identification of pollen donors requires genotyping obtained from high throughput sequencing data.…”

mentioning

confidence: 99%

Using RNA-seq to characterize pollen–stigma interactions for pollination studies

Lobaton

Andrew

Duitama

et al. 2021

Sci Rep

View full text Add to dashboard Cite

Insects are essential for the reproduction of pollinator-dependent crops and contribute to the pollination of 87% of wild plants and 75% of the world’s food crops. Understanding pollen flow dynamics between plants and pollinators is thus essential to manage and conserve wild plants and ensure yields are maximized in food crops. However, the determination of pollen transfer in the field is complex and laborious. We developed a field experiment in a pollinator-dependent crop and used high throughput RNA sequencing (RNA-seq) to quantify pollen flow by measuring changes in gene expression between pollination treatments across different apple (Malus domestica Borkh.) cultivars. We tested three potential molecular indicators of successful pollination and validated these results with field data by observing single and multiple visits by honey bees (Apis mellifera) to apple flowers and measured fruit set in a commercial apple orchard. The first indicator of successful outcrossing was revealed via differential gene expression in the cross-pollination treatments after 6 h. The second indicator of successful outcrossing was revealed by the expression of specific genes related to pollen tube formation and defense response at three different time intervals in the stigma and the style following cross-pollination (i.e. after 6, 24, and 48 h). Finally, genotyping variants specific to donor pollen could be detected in cross-pollination treatments, providing a third indicator of successful outcrossing. Field data indicated that one or five flower visits by honey bees were insufficient and at least 10 honey bee flower visits were required to achieve a 25% probability of fruit set under orchard conditions. By combining the genotyping data, the differential expression analysis, and the traditional fruit set field experiments, it was possible to evaluate the pollination effectiveness of honey bee visits under orchards conditions. This is the first time that pollen-stigma-style mRNA expression analysis has been conducted after a pollinator visit (honey bee) to a plant (in vivo apple flowers). This study provides evidence that mRNA sequencing can be used to address complex questions related to stigma–pollen interactions over time in pollination ecology.

show abstract

Transcriptome profiling of tobacco (Nicotiana tabacum) pollen and pollen tubes

Cited by 25 publications

References 47 publications

Comparative analyses of angiosperm secretomes identify apoplastic pollen tube functions and novel secreted peptides

Comparative analyses of angiosperm secretomes identify apoplastic pollen tube functions and novel secreted peptides

EXO70A2 is critical for the exocyst complex function in Arabidopsis pollen

Using RNA-seq to characterize pollen–stigma interactions for pollination studies

Contact Info

Product

Resources

About