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. 2022 Nov 10;23(22):13852.
doi: 10.3390/ijms232213852.

Comparative Seeds Storage Transcriptome Analysis of Astronium fraxinifolium Schott, a Threatened Tree Species from Brazil

Leonel Gonçalves Pereira Neto  1 , Bruno Cesar Rossini  2 , Celso Luis Marino  2   3 , Peter E Toorop  4 , Edvaldo Aparecido Amaral Silva  5
Affiliations

Comparative Seeds Storage Transcriptome Analysis of Astronium fraxinifolium Schott, a Threatened Tree Species from Brazil

Leonel Gonçalves Pereira Neto et al. Int J Mol Sci. .

Abstract

Astronium fraxinifolium Schott (Anacardiaceae), also known as a 'gonçalo-alves', is a tree of the American tropics, with distribution in Mexico, part of Central America, Argentina, Bolivia, Brazil and Paraguay. In Brazil it is an endangered species that occurs in the Cerrado, Caatinga and in the Amazon biomes. In support of ex situ conservation, this work aimed to study two accessions with different longevity (p50) of A. fraxinifolium collected from two different geographic regions, and to evaluate the transcriptome during aging of the seeds in order to identify genes related to seed longevity. Artificial ageing was performed at a constant temperature of 45 °C and 60% relative humidity. RNA was extracted from 100 embryonic axes exposed to control and aging conditions for 21 days. The transcriptome analysis revealed differentially expressed genes such as Late Embryogenesis Abundant (LEA) genes, genes involved in the photosystem, glycine rich protein (GRP) genes, and several transcription factors associated with embryo development and ubiquitin-conjugating enzymes. Thus, these results contribute to understanding which genes play a role in seed ageing, and may serve as a basis for future functional characterization of the seed aging process in A. fraxinifolium.

Keywords: differentially expressed genes; seed longevity; transcription factors.

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Conflict of interest statement

The authors declare that they have no conflict of interest.

Figures

Figure 1
Figure 1
Morphological details of species, seeds and fruits of A. fraxinifolium (A) A. fraxinifolium tree. Tree without leaves and only fruits present. The tree has a medium arboreal size, with a height of 8 to 12 m, a cylindrical and straight trunk, 60 to 80 cm in diameter at breast height (DBH) and with whitish bark; (B,C) Fruit of A. fraxinifolium adhered to the calyx. The fruit is a pseudo-samara, with a uniseriate exocarp, suberified and attached to the mesocarp; (D) Seeds from accession GOIAS, submitted to the accelerated aging test at 60% RH with zero days of aging and 100 days of accelerated aging; (E) Seeds from accession MINAS, submitted to the accelerated aging test at 60% RH with zero days of aging and 50 days of accelerated aging.
Figure 2
Figure 2
Statistical analysis of differentially expressed genes (DEGs) from A. fraxinifolium seed germination treatments. (A) Number of differentially expressed genes (DEG) in A. fraxinifolium seeds in comparison between seed treatments with and without aging (log2fold > 2, Padj ≤ 0.05). (B) Venn diagram of DEG upregulated between treatments of A. fraxinifolium seeds from MINAS (MG) × GOIAS (GO) accessions with aged seeds. (C) Venn diagram of downregulated DEG between treatments of seeds of A. fraxinifolium from accessions MINAS and GOIAS with aged seeds.
Figure 3
Figure 3
Gene ontology terms identified for DEGs in both treatments. Each color represent one main category: green for biological process, yellow for molecular function, and blue for cellular components. In the x axis the GO terms are represented.
Figure 4
Figure 4
Heatmap of selected genes possibly involved with the aging/longevity process exhibited in Table 3. Red color indicates highly expressed genes (up regulated), and green represents the down-regulated genes. The green to red color transition reflects the values of an FPKM normalized log2-transformed counts.
Figure 5
Figure 5
Gene Ontology (GO) enrichment analysis. (A,B) Enrichment analysis of DEGs that were up and down expressed in the comparison of GOIAS control vs. GOIAS aged-seeds. (C) Enrichment analysis of DEGs that were up expressed in the comparison of MINAS control vs. MINAS aged-seeds. (D,E) Enrichment analysis of DEGs that were up and down expressed in the comparison of GOIAS aged-seeds vs. MINAS aged-seeds.
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