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A proteomic investigation of sporophyte and gametophyte in the red seaweed Asparagopsis

ABSTRACT: The red seaweed, Asparagopsis taxiformis, is a promising ruminant feed-additive with significant anti-methanogenic properties, which could provide a global solution to climate-change. Emerging genomics in this species has provided a strong foundation for in-depth molecular investigations, including proteomics. Here, we investigated the A. taxiformis (L6) proteome of sporophyte and gametophyte (male and female), using soluble and insoluble extraction methods followed by mass spectrometry. A combined total of 741 and 2007 unique non-redundant proteins were identified using genome-derived and transcriptome-derived databases, respectively. Genome-derived proteins were used for further analysis including a relatively large proportion of proteins that were annotated as ion binding proteins (i.e. iron, zinc, manganese, potassium and copper), which may play a role in heavy metal bioremediation. In addition, we identified 44 photosynthesis-related proteins (e.g. phycobilisomes, photosystem I, photosystem II and ATPase) and 58 stress-related proteins (e.g. heat shock proteins and vanadium-dependent haloperoxidases). Forty proteins were predicted to be secreted, supporting a role for them in extracellular activities such as mucilage development and defence. Two Gal-2,6-Sulfurylases II proteins are likely necessary for carrageenan biosynthesis, as well as development of reproductive-associated structures. These findings provide a comprehensive overview of expressed proteins in A. taxiformis (L6) at two different life stages, highlighting the potential of proteomic approaches and warrant further studies for understanding protein functions.

INSTRUMENT(S): X500R QTOF

ORGANISM(S): Asparagopsis Taxiformis

TISSUE(S): Whole Body

SUBMITTER: Tianfang Wang

LAB HEAD: Tianfang Wang

PROVIDER: PXD035669 | Pride | 2023-01-26

REPOSITORIES: Pride

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Project description:Purpose: The plant hormone abscisic acid (ABA) coordinates responses leading to desiccation tolerance in all land plants. RNA-seq was employed to obtain differential expression data in response to ABA, dehydration and mannitol treatments in wild-type tissue. This was compared against the targeted gene knockout line PpanrKO. The ANR gene encodes a novel ABA regulator identified from ABA non-responsive (anr) mutant screens. Methods: RNA-seq on the Illumina HiSeq platform and mapping to the V3.0 genome and transcriptome assemblies using tophat2 enabled gene abundance data to be generated for differential gene expression analysis with edgeR in the Trinity package. Results: We observe a coordinated up-regulation of ca. 900 genes, with extensive overlaps between hormone and stress treatments as well as ca. 900 down-regulated genes. Of the up-regulated genes, those encoding LEA proteins, membrane channel proteins and transporters and oxidative-stress-associated genes comprised a significant fraction while those encoding growth-associated cell wall enzymes (e.g. expansins) and components of secondary metabolism (notably enzymes in the phenylpropanoid pathway) were prominent in the down-regulated genes. The ANR gene, a modular MAP3 kinase comprising an N-terminal PAS domain, a central EDR domain and a C-terminal MAPKKK-like domain unique to basal plant lineages, is found to play a central role in these responses with mutants failing to accumulate the wide spectrum of dehydration tolerance-associated gene products characteristic of the wild-type response and do not acquire ABA-dependent desiccation tolerance. Conclusions: Our study adds considerable data to the stress responses in bryophytes helping understand the mechanisms behind vegetative dehydration tolerance; an important trait for crop species. We also confirmed the vital role that PpANR plays in these responses in the moss Physcomitrella patens likely representing an ancestral mechanism for ABA-mediated dehydration tolerance vital in the conquest of land. RNA from 3 biological replicas was bulked per library. 8 libraries were generated from two genotypes, wild-type and PpanrkO, which included 3 treatments and a control sample for each.

			Action	DRS
	Female_TrisAcetone.mzxml	Mzxml
	Female_TrisAcetone.xlsx	Xlsx
	Female_TrisBot.mzxml	Mzxml
	Female_TrisBot.xlsx	Xlsx
	Female_TrisTop.mzxml	Mzxml

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A proteomic investigation of sporophyte and gametophyte in the red seaweed Asparagopsis

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