Hippeastrum species in areas of restinga in the state of Rio de Janeiro, Brazil: pollen characters

Candido, Renata Suzano; Fourny, Ana Carolina da Silva; Gonçalves-Esteves, Vânia; Lopes, Rosana Conrado

doi:10.1590/s0102-33062013000400003

Cited by 6 publications

(7 citation statements)

References 18 publications

(32 reference statements)

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“…First, we found a large interspecific variation in extracted DNA yields. Possible causes of this variation may be interspecific differences in pollen wall structure38, pollen size394041, genome size42, the number of marker copies and DNA extraction efficiency from protoplasts43. Therefore, in routine studies these between-species variations will require calibration, for each focal species, of the relationship between the number of pollen grains and the number of sequences.…”

Section: Discussionmentioning

confidence: 99%

Using metabarcoding to reveal and quantify plant-pollinator interactions

Pornon

Escaravage

Burrus

et al. 2016

Sci Rep

127

173

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Given the ongoing decline of both pollinators and plants, it is crucial to implement effective methods to describe complex pollination networks across time and space in a comprehensive and high-throughput way. Here we tested if metabarcoding may circumvent the limits of conventional methodologies in detecting and quantifying plant-pollinator interactions. Metabarcoding experiments on pollen DNA mixtures described a positive relationship between the amounts of DNA from focal species and the number of trnL and ITS1 sequences yielded. The study of pollen loads of insects captured in plant communities revealed that as compared to the observation of visits, metabarcoding revealed 2.5 times more plant species involved in plant-pollinator interactions. We further observed a tight positive relationship between the pollen-carrying capacities of insect taxa and the number of trnL and ITS1 sequences. The number of visits received per plant species also positively correlated to the number of their ITS1 and trnL sequences in insect pollen loads. By revealing interactions hard to observe otherwise, metabarcoding significantly enlarges the spatiotemporal observation window of pollination interactions. By providing new qualitative and quantitative information, metabarcoding holds great promise for investigating diverse facets of interactions and will provide a new perception of pollination networks as a whole.

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Section: Discussionmentioning

confidence: 99%

Using metabarcoding to reveal and quantify plant-pollinator interactions

Pornon

Escaravage

Burrus

et al. 2016

Sci Rep

127

173

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“…CHR has small (20 µm to 35 µm diameter), echinate pollen grains with a tectate ectoexine with large lacunae and three apertures 38 , and a 2 C value of 11.87 pg DNA 36 . HIP pollen grains are bigger (50 µm to 100 µm in diameter), with a semitectate ectoexine and one aperture 35 . Their DNA content ranges between 2 C = 13.35 pg and 2 C = 17.09 pg 42 .…”

Section: Methodsmentioning

confidence: 99%

“…-Asteraceae, and Hippeastrum sp. -Amaryllidaceae) of which the pollen has very different characteristics (size, exin structure, DNA content) [35][36][37][38][39] . We analysed two plant specific loci, the internal transcribed spacer region 1 (ITS1, ≈ 300 bp) of the nuclear ribosomal region, and the plastidial P6-loop of trnL (UAA) intron (≈75 bp), efficient for investigating potential degraded DNA, respectively, hereafter referred to as ITS1 and trnL.…”

mentioning

confidence: 99%

Experimental quantification of pollen with DNA metabarcoding using ITS1 and trnL

Baksay

Pornon

Burrus

et al. 2020

Sci Rep

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Although the use of metabarcoding to identify taxa in DNA mixtures is widely approved, its reliability in quantifying taxon abundance is still the subject of debate. In this study we investigated the relationships between the amount of pollen grains in mock solutions and the abundance of highthroughput sequence reads and how the relationship was affected by the pollen counting methodology, the number of PCR cycles, the type of markers and plant species whose pollen grains have different characteristics. We found a significant positive relationship between the number of DNA sequences and the number of pollen grains in the mock solutions. However, better relationships were obtained with light microscopy as a pollen grain counting method compared with flow cytometry, with the chloroplastic trnL marker compared with ribosomal ITS1 and with 30 when compared with 25 or 35 PCR cycles. We provide a list of recommendations to improve pollen quantification. Environmental DNA metabarcoding is a molecular method that consists of investigating environmental DNA samples made of complex mixtures of genomes from numerous organisms 1. Due to new sequencing technologies and bioinformatics tools, metabarcoding has been increasingly used to identify taxa in environmental samples 1 to monitor biodiversity 2-4 , to investigate ecosystem functioning 5 and interaction networks 6-8 , in both aquatic and terrestrial ecosystems. Nevertheless, its reliability in quantitative approaches, which depend on the match between counts of high-throughput sequence reads and the amount of sampled biological material 2 , is still the subject of debate 9,10. While taxon identification can reveal individual diet breadth 11 , species richness, and the composition of habitats 2 , communities 12 and ecological networks 4 , taxon quantification provides knowledge on species evenness in those habitats, communities and diets or on the level of individual or species specialization in networks, all of which is very useful in ecological studies. Research on pollination and knowledge of the quantities of pollen transported by pollinators allow for the estimation of plant-pollinator interaction strength and hence it gives a more realistic representations of networks than those made possible using traditional approaches such as observing visits to plants by pollinators 9,13. Metabarcoding has been used in pollen studies to identify pollen in honey 14-16 , insect loads 6-8,17 , insect nests 18 , airborne samples 19 , and to quantify pollen abundance across various sample types. Several studies found significant positive relationships between pollen abundance (estimated using light microscopy) or pollen DNA quantities, and the abundance or the frequencies of high-throughput sequencing reads in experimental samples 10,16,17,20,21 , airborne samples 22,23 , insect pollen loads 21,24-27 or in brood cells of solitary bees 28. Conversely, other studies found low or no significant pollen-sequence abundance relationships when using ITS2 markers applied to pollen provision in...

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“…The Amaryllidaceae, a plant family in the monocot order Asparagales, is composed of three subfamilies: Amaryllidoideae, Agapanthoideae and Allioideae [ 6 , 7 ]. The Amaryllidoideae include about 80 genera [ 8 ] that are mainly distributed in tropical and subtropical regions, but also in temperate zones [ 9 ]. A particular feature of plants of the Amaryllidoideae subfamily is their content of an exclusive, numerous and still expanding group of alkaloids, known as Amaryllidaceae alkaloids.…”

Section: Introductionmentioning

confidence: 99%

Hippeastrum reticulatum (Amaryllidaceae): Alkaloid Profiling, Biological Activities and Molecular Docking

et al. 2017

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The Amaryllidaceae family has proven to be a rich source of active compounds, which are characterized by unique skeleton arrangements and a broad spectrum of biological activities. The aim of this work was to perform the first detailed study of the alkaloid constituents of Hippeastrum reticulatum (Amaryllidaceae) and to determine the anti-parasitological and cholinesterase (AChE and BuChE) inhibitory activities of the epimers (6α-hydroxymaritidine and 6β-hydroxymaritidine). Twelve alkaloids were identified in H. reticulatum: eight known alkaloids by GC-MS and four unknown (6α-hydroxymaritidine, 6β-hydroxymaritidine, reticulinine and isoreticulinine) by NMR. The epimer mixture (6α-hydroxymaritidine and 6β-hydroxymaritidine) showed low activity against all protozoan parasites tested and weak AChE-inhibitory activity. Finally, a molecular docking analysis of AChE and BuChE proteins showed that isoreticulinine may be classified as a potential inhibitory molecule since it can be stabilized in the active site through hydrogen bonds, π-π stacking and hydrophobic interactions.

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Hippeastrum species in areas of restinga in the state of Rio de Janeiro, Brazil: pollen characters

Cited by 6 publications

References 18 publications

Using metabarcoding to reveal and quantify plant-pollinator interactions

Using metabarcoding to reveal and quantify plant-pollinator interactions

Experimental quantification of pollen with DNA metabarcoding using ITS1 and trnL

Hippeastrum reticulatum (Amaryllidaceae): Alkaloid Profiling, Biological Activities and Molecular Docking

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