Application of proteomics to the study of pollination drops

Prior, Natalie; Little, Stefan A.; Pirone, Cary; Gill, Julia E.; Smith, Derek; Han, Jun; Hardie, Darryl B.; O’Leary, Stephen; Wagner, Rebecca E.; Cross, Tyra J.; Coulter, Andrea; Borchers, Christoph H.; Olafson, Robert W.; Aderkas, Patrick von

doi:10.3732/apps.1300008

Cited by 13 publications

(9 citation statements)

References 29 publications

(48 reference statements)

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“…The sampling was performed from 1930–2130 h during peak pollination times [14,21], although the pollination drops were sometimes additionally collected from 0830–1030h when the ambient humidity exceeded 90%. Our drop sampling strategy follows the methods applied in previous studies [30,31,33,34,35,63,64]. The female and male drops of G. gnemon/G.…”

Section: Methodsmentioning

confidence: 99%

Pollination Drop Proteome and Reproductive Organ Transcriptome Comparison in Gnetum Reveals Entomophilous Adaptation

Hou

Saunders

Deng³

et al. 2019

Genes

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Gnetum possesses morphologically bisexual but functionally unisexual reproductive structures that exude sugary pollination drops to attract insects. Previous studies have revealed that the arborescent species (G. gnemon L.) and the lianoid species (G. luofuense C.Y.Cheng) possess different pollination syndromes. This study compared the proteome in the pollination drops of these two species using label-free quantitative techniques. The transcriptomes of fertile reproductive units (FRUs) and sterile reproductive units (SRUs) for each species were furthermore compared using Illumina Hiseq sequencing, and integrated proteomic and transcriptomic analyses were subsequently performed. Our results show that the differentially expressed proteins between FRUs and SRUs were involved in carbohydrate metabolism, the biosynthesis of amino acids and ovule defense. In addition, the differentially expressed genes between the FRUs and SRUs (e.g., MADS-box genes) were engaged in reproductive development and the formation of pollination drops. The integrated protein-transcript analyses revealed that FRUs and their exudates were relatively conservative while the SRUs and their exudates were more diverse, probably functioning as pollinator attractants. The evolution of reproductive organs appears to be synchronized with changes in the pollination drop proteome of Gnetum, suggesting that insect-pollinated adaptations are not restricted to angiosperms but also occur in gymnosperms.

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Section: Methodsmentioning

confidence: 99%

Pollination Drop Proteome and Reproductive Organ Transcriptome Comparison in Gnetum Reveals Entomophilous Adaptation

Hou

Saunders

Deng³

et al. 2019

Genes

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“…1 ). These minute drops of liquid, 10–1000 nL, play a critical role in gymnosperm reproduction (Prior et al 2013 ). Pollination drops receive pollen and move it to the ovule interior where it germinates (Gelbart and von Aderkas 2002 ).…”

Section: Introductionmentioning

confidence: 99%

Complex reproductive secretions occur in all extant gymnosperm lineages: a proteomic survey of gymnosperm pollination drops

et al. 2018

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Key message Complex protein-containing reproductive secretions are a conserved trait amongst all extant gymnosperms; the pollination drops of most groups include carbohydrate-modifying enzymes and defence proteins. Abstract Pollination drops are aqueous secretions that receive pollen and transport it to the ovule interior in gymnosperms (Coniferales, Cycadales, Ginkgoales, Gnetales). Proteins are well established as components of pollination drops in conifers (Coniferales) and Ephedra spp. (Gnetales), but it is unknown whether proteins are also present in the pollination drops of cycads (Cycadales), Ginkgo (Ginkgoales), Gnetum (Gnetales), or in the pollination drops produced by sterile ovules occurring on pollen plants in the Gnetales. We used liquid chromatography–tandem mass spectrometry followed by database-derived protein identification to conduct proteomic surveys of pollination drops collected from: Ceratozamia hildae , Zamia furfuracea and Cycas rumphii (Cycadales); Ginkgo biloba (Ginkgoales); Gnetum gnemon and Welwitschia mirabilis , including pollination drops from both microsporangiate and ovulate plants (Gnetales). We identified proteins in all samples: C. hildae (61), Z. furfuracea (40), C. rumphii (9), G. biloba (57), G. gnemon ovulate (17) and sterile ovules from microsporangiate plants (25) and W. mirabilis fertile ovules (1) and sterile ovules from microsporangiate plants (138). Proteins involved in defence and carbohydrate modification occurred in the drops of most groups, indicating conserved functions for proteins in pollination drops. Our study demonstrates that all extant gymnosperm groups produce complex reproductive secretions containing proteins, an ancient trait that likely contributed to the evolutionary success of seed plants. Electronic supplementary material The online version of this article (10.1007/s00497-018-0348-z) contains supplementary material, which is available to authorized users.

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“…The existence of proteins in the PDs of many gymnosperms has been reported recently, and the identified proteins strongly indicate that the biological roles of PDs might be more complex than simply the receipt and transportation of pollen during pollination. [31][32][33] The number of proteins varies from species to species in gymnosperms PDs. For instance, the number of characterized proteins in degradome and secretome of Ephedra species PDs ranged from 6 to 20, 32 which were less than that of G. biloba.…”

Section: Discussionmentioning

confidence: 99%

Constituent analysis and proteomic evaluation of ovular secretions inGinkgo biloba: not just a pollination medium

Cheng

Zhao

Jiang

et al. 2018

Plant Signaling & Behavior

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Pollination drop (PD) is a characteristic feature of major wind-pollinated gymnosperms and plays a vital role during the course of pollination, however, the composition and proteomic profile of PDs in Ginkgo biloba remain unclear. Through inductively coupled plasma mass spectrometry, we detected mineral elements in PDs, including calcium (Ca), sulfur (S), magnesium, boron, and potassium (K), among which S, Ca, and K were found at high levels. The total sugar concentration was approximately 5.908 mg/mL, which accounted for approximately 5.9% (mass ratio) of the PD. The sugars primarily consisted of fructose, glucose, and sucrose, of which the glucose level was highest, accounting for 57.6%, followed by fructose (37.1%) and sucrose (5.3%). We also used FTIR to validate the presence of sugars and proteins in PDs. Further proteomic analysis revealed that the PD contained calmodulin, α-L-arabinofuranosidase, β-D-xylosidase, superoxide dismutase, α-L-arabinosidase, glutathione S-transferase, histones, glycine-rich family protein, methionine synthase, and arabinogalactan, suggesting that proteins present in PDs of G. biloba play a critical role in the defense against external bacteria and facilitate germination and growth of the pollen tube. Our results suggest that PDs are not merely a medium to receive and transport pollen but may also play a more complex biological role in pollination and fertilization.

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Application of proteomics to the study of pollination drops

Cited by 13 publications

References 29 publications

Pollination Drop Proteome and Reproductive Organ Transcriptome Comparison in Gnetum Reveals Entomophilous Adaptation

Pollination Drop Proteome and Reproductive Organ Transcriptome Comparison in Gnetum Reveals Entomophilous Adaptation

Complex reproductive secretions occur in all extant gymnosperm lineages: a proteomic survey of gymnosperm pollination drops

Constituent analysis and proteomic evaluation of ovular secretions inGinkgo biloba: not just a pollination medium

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