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Overall, 288 genes were
identified as putative AP2/ERF genes in the sunflower genome. The predicted HaAP2/ERF(generic
name and locus Tag has been showed in Table S in details) genes were then chosen
based on their chromosome location and family classification (Table S1). Based
on the classification, we are grouped of them into21 ANT, 14 AP2, 4 RAV, 1
Soloist, 105 DREB, and 143ERF. 35 genes AP2 TFs contain two AP2 domains or a
single AP2 domain that is similar to the AP2 domains in the double domain
groups were assigned to the AP2 family, and 4 genes attracted a single AP2/ERF
DNA binding motif together with a B3 type domain were grouped into the RAV
family. The 248 genes have only one domino belonging to the ERF subfamily,
which itself divides into subgroups of ERF and DREB. In addition, a specific
gene called HaAP2/ERF-288 has a similar resemblance to other family
members that are in the subgroup Soloist (Table S2).

Chromosome distribution
analysis found that theHaAP2/ERF genes were unevenly distributed on all
of 17 chromosomes of sunflower (Fig. 1). In detail, 30 AP2/ERF genes located on
the chromosome 2 and 16 which have the maximum number of AP2/ERFs, while there were only 9genes on the
chromosome5, which have the minimum number of AP2/ERFs. The putative proteins
of HaAP2/ERFs ranged from 78 to 1347 amino acids in length; with
molecular weights (MW) ranging from 8.52 to 69.44kDa and theoretical
isoelectric points (PI) ranging from 4.48 to10.27. Subcellular localization analysis
indicated that majority of HaAP2/ERFs (234 out of 288, 81.25 %)
localized in the nucleus, while 54 genes were located in the extracellular
(Table S1).

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Phylogenetic relationship, conserved
motif and gene structure analysis


To assess
the evolutionary associations of the HaAP2/ERF genes, phylogenetic
analysis was performed based on the multiple correlation of all HaAP2/ERF
with the ArabidopsisAP2/ERF genes. The phylogenetic tree categorizes all of the
AP2/ERF genes into three main categories (ERF, AP2 and RAV) depending on the
composition of their amplitude as described above (Fig. 2). A non-rooted
phylogenetic tree was made with HaAP2/ERF family proteins (Fig. 3). In
addition, ERF clades were further divided into 10 groups. According to Arabidopsisclassification
criteria6, ERFs families can be further subdivided into subgroup of DREB
and ERF families. Four groups (I-IV) belong to the DREB, while the remaining
six groups (V-X) were ERF subgroups (Fig. 4). It was found that DREB subgroups are
major contribute to plant abiotic stress responses, and many of the DREBs that
have been tolerated by stress from many plants have been recognized to date24,25,48.
Identification of DREB genes of Helianthus annuus provides valuable
resources for the characterization of stress-response genes. In addition,
bootstrapping values of the nodes in phylogenetic tree were not very high in
each class, which was consistent with earlier studies6,49. The
reliability of the NJ tree was confirmed

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