Molecular Plant Breeding 2015, Vol.6, No.20, 1
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results showed that 12 ESTs might be related to signal
transduction (Table 5).
Ubiquitin-conjugated E2 enzyme (UBE2) is one of the
main components of the proteasome degradation
cascade. Previous studies have shown an increase of
expression levels in individuals challenged to some
pathogen organism such as virus and bacteria. In the
process of the ubiquitination, the main enzymes
contain ubiquitin-activating enzyme E1, ubiquitin-
conjugating enzyme E2 and ubiquitin-protein ligase
E3. These enzymes play an important role in
maintaining the balance of generation and degradation
of intracellular protein, the cell homeostasis and
normal function, and so on (Markson et al., 2009).
UBE2 protein has a very important relationship with
stress resistance. Drought, High temperature, heavy
metals and other abiotic stress can induce the
expression of ubiquitin-conjugating enzyme E2 in
plant, which in turn enhances the adaptive ability to
adverse environment.
There were about ten ESTs that may relate to signal
trusduction showed in table 5. We compared and
analyzed these genes with Banana Genome Hub,
Genbank, and KEGG Pathway, but we can’t get clear
conclusion about how the banana carried out the
signal transduction responded to Fusarium wilt
disease infection.
Gene expression verification by sqRT-PCR
We choose six ESTs belonged to different functional
catalogues to analyze the expression patterns (Table 6)
by sqRT-PCR to validate the reliability of cDNA-
AFLP experiment, and quantitatively assessed the
relative abundance of ESTs in banana leaf between the
control and infection treatments at four different time
points (Figure 4). As results, the quantitative expression
patterns (increase or decrease in RNA level) of the 6
ESTs were similar in both cDNA-AFLP and sqRT-
PCR experiment, it indicated that cDNA-AFLP was an
effective method to identify differential expressed genes.
Discussion
In this study, the interaction mechanism was done by
cDNA-AFLP. After analysis of cDNA-AFLP, 223
differentially expressed fragments were obtained.
Sequencing and analyzing these fragments, 137 kinds
Table 5 ESTs related to signal transduction
NO.
GenBank ID of homology
Putative function
E10
ACG24192.1
structure-specific recognition protein 1
E67
NP_001149824.1
ADP-ribosylation factor GTPase-activating protein 3
E87
NP_001147943.1
signal transducer, WD40 domain
E106
ZP_08497599.1
membrane-associated diguanylate cyclase
E143
XP_001607074.2
metabotropic glutamate receptor 7
E154
XP_003613587.1
Blue-light-activated histidine kinase
E184
YP_002131246.1
TonB-dependent receptor
E187/ E189
CBY17542.1
Ras-related nuclear proteins (Ran)/TC4 family of small GTPases
E200/ E202
NP_001146868.1
ubiquitin-conjugating enzyme E2, catalytic (UBCc)domain
E214
BAC05850.1
seven transmembrane helix receptor (rhodopsin family)
Table 6 The ESTs verified by sqRT-PCR
NO.
GenBank
ID
of
homology
Length (bp)
Putative function
Catalogue
E87
NP_001147943.1
262
signal transducer
Signal transduction
E96
296
New sequence
New EST
E108
AM183222.2
278
cytochrome c biogenesis protein
Primary Metabolism
E142
ABF94667.1
169
expressed protein
unknown
E220
XP_003606916.1
137
Glycine-rich RNA binding protein
Transcription
E205
211
New sequence
New EST
