Dataset Information

Analysis of Pollen Allergens in Lily by Transcriptome and Proteome Data.

ABSTRACT: The lily (Lilium spp.) anther contains a lot of pollen. It is not known if lily pollen contains allergens, and therefore screening pollen allergy-related proteins and genes is necessary. The pollen development period of lily 'Siberia' was determined by microscope observation. Early mononuclear microspores and mature pollens were used as sequencing materials. The analysis of the pollen transcriptome identified differentially expressed genes (DEGs), e.g., Profilin, Phl p 7 (Polcalcin), Ole e 1, and Phl p 11, which are associated with pollen allergens. The proteome analysis positively verified a significant increase in pollen allergenic protein content. The expression levels of LoProfiilin and LoPolcalcin, annotated as allergen proteins, gradually increased in mature pollen. LoProfiilin and LoPolcalcin were cloned and their open reading frame lengths were 396 bp and 246 bp, which encoded 131 and 81 amino acids, respectively. Amino acid sequence and structure alignment indicated that the protein sequences of LoProfilin and LoPolcalcin were highly conserved. Subcellular localization analysis showed that LoProfilin protein was localized in the cell cytoplasm and nucleus. LoProfilin and LoPolcalcin were highly expressed in mature pollen at the transcriptional and protein levels. A tertiary structure prediction analysis identified LoProfilin and LoPolcalcin as potential allergens in lily pollen.

SUBMITTER: Feng J

PROVIDER: S-EPMC6929097 | biostudies-literature |

REPOSITORIES: biostudies-literature

ACCESS DATA

Similar Datasets

Project description:BackgroundRespiratory allergy triggered by pollen allergens is increasing at an alarming rate worldwide. Sunflower pollen is thought to be an important source of inhalant allergens. Present study aims to identify the prevalence of sunflower pollinosis among the Indian allergic population and characterizes the pollen allergens using immuno-proteomic tools.MethodologyClinico-immunological tests were performed to understand the prevalence of sensitivity towards sunflower pollen among the atopic population. Sera from selected sunflower positive patients were used as probe to detect the IgE-reactive proteins from the one and two dimensional electrophoretic separated proteome of sunflower pollen. The antigenic nature of the sugar moiety of the glycoallergens was studied by meta-periodate modification of IgE-immunoblot. Finally, these allergens were identified by mass-spectrometry.ResultsPrevalence of sunflower pollen sensitization was observed among 21% of the pollen allergic population and associated with elevated level of specific IgE and histamine in the sera of these patients. Immunoscreening of sunflower pollen proteome with patient sera detected seven IgE-reactive proteins with varying molecular weight and pI. Hierarchical clustering of 2D-immunoblot data highlighted three allergens characterized by a more frequent immuno-reactivity and increased levels of IgE antibodies in the sera of susceptible patients. These allergens were considered as the major allergens of sunflower pollen and were found to have their glycan moiety critical for inducing IgE response. Homology driven search of MS/MS data of these IgE-reactive proteins identified seven previously unreported allergens from sunflower pollen. Three major allergenic proteins were identified as two pectate lyases and a cysteine protease.ConclusionNovelty of the present report is the identification of a panel of seven sunflower pollen allergens for the first time at immuno-biochemical and proteomic level, which substantiated the clinical evidence of sunflower allergy. Further purification and recombinant expression of these allergens will improve component-resolved diagnosis and therapy of pollen allergy.

Project description:Pollen germination is a crucial process in the life cycle of flowering plants, signifying the transition of quiescent pollen grains into active growth. This study delves into the dynamic changes within organelles and the pivotal role of autophagy during lily pollen germination. Initially, mature pollen grains harbor undifferentiated organelles, including amyloplasts, mitochondria, and the Golgi apparatus. However, germination unveils remarkable transformations, such as the redifferentiation of amyloplasts accompanied by starch granule accumulation. We investigate the self-sustained nature of amylogenesis during germination, shedding light on its association with osmotic pressure. Employing BODIPY 493/503 staining, we tracked lipid body distribution throughout pollen germination, both with or without autophagy inhibitors (3-MA, NEM). Typically, lipid bodies undergo polarized movement from pollen grains into elongating pollen tubes, a process crucial for directional growth. Inhibiting autophagy disrupted this essential lipid body redistribution, underscoring the interaction between autophagy and lipid body dynamics. Notably, the presence of tubular endoplasmic reticulum (ER)-like structures associated with developing amyloplasts and lipid bodies implies their participation in autophagy. Starch granules, lipid bodies, and membrane remnants observed within vacuoles further reinforce the involvement of autophagic processes. Among the autophagy inhibitors, particularly BFA, significantly impede germination and growth, thereby affecting Golgi morphology. Immunogold labeling substantiates the pivotal role of the ER in forming autophagosome-like compartments and protein localization. Our proposed speculative model of pollen germination encompasses proplastid differentiation and autophagosome formation. This study advances our understanding of organelle dynamics and autophagy during pollen germination, providing valuable insights into the realm of plant reproductive physiology.

Project description:BackgroundPollen contains hundreds of different proteins. However, only a small fraction of them have been identified to be allergenic. We aimed to test the hypothesis that most pollen proteins are non-allergenic due to their high extent of sequence conservation among non-related species.MethodsData on the composition of pollen proteomes of birch (Betula pendula), pellitory (Parientaria judaica) and timothy grass (Phleum pratense) were obtained from the literature. Sequences were downloaded from UniProt and manually classified into allergens and non-allergens. Complete proteome sequences of 3 dicotyledonous species (Arabidopsis thaliana, Populus trichocarpa and Vitis vinifera), 2 monocotyledons (Oryza sativa subsp. japonica and Zea mays) and one moss (Physcomitrella patents) were downloaded from ENSEMBL Plants. Sequences of pollen proteins were compared to these proteomes by using BLAST and the hits yielding the highest sequence identity recorded taking into account only sequence alignments at least 40 residues in length. The distributions of maximum sequence identities of allergens and non-allergens from each species were compared using the Mann-Whitney test.ResultsAllergens from birch and pellitory pollen were significantly (P < 0.001) less similar to proteins from monocots than non-allergenic pollen proteins. Median sequence identities to the nearest rice and maize homologues were 49 and 52% for birch allergens, 86 and 85% for birch non-allergens, 37 and 37% for pellitory allergens, and 87 and 89% for pellitory non-allergens. Similarly, timothy grass pollen allergens were significantly (P < 0.0001) less similar to dicot proteins than non-allergenic pollen proteins. Median sequence identities to the nearest homologues were 43 to 44% for allergens and 81 to 83% for non-allergens. A comparison of all 3 pollen proteomes to sequences from the moss P. patens yielded similarly significant differences.ConclusionsPollen allergens belong to evolutionary less conserved protein families than non-allergenic pollen proteins. The continual exposure of the human immune system to nearly identical and hence highly cross-reactive conserved proteins from multiple pollen and plant food species most likely leads to the induction of immunological tolerance rather than allergic sensitization.

Dataset Information

Analysis of Pollen Allergens in Lily by Transcriptome and Proteome Data.

Similar Datasets

OmicsDI is part of the ELIXIR infrastructure

Tweets