Ossman Barrientos-Díaz, Nicole Moreira Veto, Franceli R. Kulckeski, Alexandra Antunes Mastroberti , Andreia C. Turchetto-Zolet

1. GENP
Glycerol-3-phosphate Acyltransferase (GPAT)
genes of Eugenia uniflora L. and its potential
involvement in adaptation mechanisms
Ossman Barrientos-Díaz1*, Nicole Moreira Veto1, Franceli R. Kulckeski2,
Alexandra Antunes Mastroberti3 , Andreia C. Turchetto-Zolet1*
1. Programa de Pós-Graduação em Genética e Biologia Molecular, Departamento de Genética, Instituto de Biociências,
Universidade Federal do Rio Grande do Sul (UFRGS), Brasil.
2. Programa de Pós-Graduação em Biologia Celular e do Desenvolvimento, Departamento de Biologia Celular, Embriologia e
Genética (UFSC), Brasil
3. Programa de Pós-Graduação em Botânica, Laboratório de Anatomia vegetal, Instituto de Biociências, UFRGS, Brasil.
*ossmanbarrientos@ufrgs.br; aturchetto@gmail.com - OssmanBD

2. Eugenia uniflora L. (Myrtaceae) is a versatile
species that occurs in heterogeneous
environments within Atlantic Forest domain
(Figure 1). The species is popularly known as
pitanga or Brazilian cherry and has been
proposed to be a good model to study adaptive
evolution. (Turchetto-Zolet et al., 2016).
BACKGROUND
Figure 2. Scheme for triacylglycerol (TAG) biosynthesis via the
glycerol phosphate pathway, adapted from Körbes et al. (2016)
The main goal of this study was to identify and characterize genes that encoding GPATs in
E. uniflora and to examine the potential involvement of lipids in adaptation mechanisms of
this species
Lipids may offer a response to adaptation given that are composed of several types of
fatty acids and their derivatives, such as lipid polyesters, glycerolipids and sterols. (Chen et al.,
2011)
Glycerol-3-Phosphate Acyltransferase (GPAT) is the first gene family to participate in
Kennedy pathway (Figure 2), the major pathway for lipid biosynthesis (Waschburger et al., 2018).
These genes are related to the morphological and functional evolution of terrestrial plants,
as well as adaptation processes, due to their relationship with the stress response in plants
(Maraschin et al., 2019)
Figure 1. Geographical distribution and Haplotype diversity of E.
uniflora, adapted from Turchetto et al. (2016)

3. MATERIAL AND METHODS
IDENTIFICATION AND IN SILICO CHARACTERIZATION OF E. uniflora GPATs
1
A
Eugenia uniflora TRANSCRIPTOME
Germination
RNA Extraction
CTAB method
ZYMO Direct-zol™
RNA MiniPrep
▪ Safranin and Astra-Blue
Bukatsch (1972)
▪ Sudan III (Sass, 1951)
▪ Black sudan (Pearse, 1980)
▪ Nile Blue (Cain, 1947)
Anatomical and histochemical analysis of leaves
275
225
Plant Material
B

4. After alignments and phylogenetic analysis, we identified seven putative GPATs in E. uniflora,
orthologs of GPAT1, GPAT2/3, GPAT6, GPAT4/8, and GPAT9 (Fig. 3). Interestingly, there are no
clustered sequences with GPAT 5/7.
80% seed germination
(Amador & Barbedo, 2015; Carvalho et al., 1998)
Figure 4. Evolutionary relationships among GPAT genes 1-8
(A) and 9 (B) of Rosidae clade and Eugenia uniflora (Euni).
Figure 6. Structural and histochemical characteristics of two populations of E. uniflora. (A-H) Restinga and
(I-P) Riparian forest. (B, I) Astra Blue and Safranin test, cross section. Sudan III (C, L) Control (D, K)
Samples. Nile Blue (E, N) Control, (F, M) samples. Black Sudan (G, P) Control, (H, O) samples. C (cuticle),
(cr) crystal, (ec) epidermis cell, (pp) palisade parenchyma. Scale of all the images correspond to 20µm
The histochemical tests
(Sudan III, Sudan Black,
and Nile Blue) showed
that the presence of
lipid compounds
(probably essential oils,
characteristic of the
Myrtaceae family) was
evidenced in secretory
cavities, parenchyma
cells and especially in
the cuticle of the
individuals of E.
uniflora (Figure 6).
The gene ontology and BLAST analyses revealed
contigs with similarity to GPAT genes of Eucalyptus
grandis and Arabidopsis thaliana (Table 1).
Figure 5. Eugenia uniflora plants cultivated in greenhouse. The seeds used
for germination in greenhouse were obtained from (green) Restinga
population (RE-RJ) and (red) from Riparian Forest population (RF-RS).
RE
RF
Figure 3. Evolutionary relationships among GPAT genes of
Arabidopsis thaliana (Ath), Eucalyptus grandis (Eucgr) and
Eugenia uniflora (Euni).
Selection of candidate genes of GPAT
RESULTS
GPAT 4-8
GPAT 6
GPAT 5-7
GPAT 1-3
GPAT 9
A
B
Anatomical and histochemical analysis

5. Some parameters on the GERMINATION of the seeds of
E. uniflora were calculated (Table 3).
Despite the knowledge that there is still little about the functions of GPATs in plants and more specifically in native
plants, the study allows to obtain a light on those mechanisms that participate in the adaptive evolution.
These results reinforce and offer a further path that allows us to explore more deeply the importance of the lipids in
the adaptation processes of native species.
Thus, our study was supported by morphological and histochemical analyzes suggesting that the anatomy of the
populations is similar but the accumulation of lipids varies when compared to the dimensions of the leaves.
Analysis of expression profile of the GPAT genes under stress conditions in different types of controlled
environments, as well as the positive selection analysis.
DISCUSSION AND CONCLUSION
Phylogenetics reconstructions (Fig. 4a, b) DUPLICATION
AND DIVERSIFICATION events of the different GPAT
orthologs of E. uniflora are appreciated.
Phylogenetic analyzes indicate that the SEVEN GPAT GENES recovered
from E. uniflora were grouped into several subclades of the gene family.
MORPHOLOGICAL AND HISTOCHEMICAL ANALYZES
suggesting that the anatomy of the populations is similar.
The importance of the lipids in the adaptation
processes of NATIVE SPECIES.
SUPPORT FINANCEMENT:
Plants that growth in the greenhouse recovered and expressed the
same PHENOTYPE of the localities where they were collected (Fig. 4)
CUTIN in leaves in different volumes is a possible mechanism
of phenotypic plasticity of the species