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Genomics and Applied Biology, 2010, Vol.1 No.2
http://gab.sophiapublisher.com
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Figure 3 Multiple sequence alignments of
HD
homologs
Note: The two conserved residues of
A. hypogaea
HD corresponding to
E. coli
FabZ His54, Glu68 and
E. coli
FabA His70, Asp84
were highlighted in asterisks; GenBank accession numbers (locus tags) for FabZ were respectively as follows:
Arachis hypogaea
(FJ768727),
Glycine max
(ACU23629),
Arabidopsis thaliana
(NP_565528),
Synechocystis
PCC6803 (NP_441227),
Chlorella
NC64A
(ChlNC64A_50849),
Escherichia coli
(AAC36917); GenBank accession numbers (locus tags) for FabA were respectively as
follows:
Escherichia coli
(NP_415474),
Klebsiella pneumoniae
(YP_001334649),
Haemophilus influenzae
(NP_439476),
Azotobacter vinelandii
(YP_002800049), and
Pseudomonas aeruginosa
(AAG04999)
1.2 Quantitative real-time RT-PCR analysis
The quantitative real-time RT-PCR (qRT-PCR) was
employed to confirm the expression patterns of
AhKR
,
AhHD
, and
AhENR
genes in four peanut tissues and at
different developmental stages of seeds. β
-
actin was
used as an internal reference control for total RNA
input. β
-
actin PCR product was not detected when
reverse transcriptase was omitted, indicating that the
RNA template was free of genomic DNA. The results
revealed that the three genes were expressed
dominantly in leaf among four tissues tested, and
expressed at the lowest level in stem (Figure 7). In
addition, the expression level of these genes in seed
was also relatively high. The expression patterns of
the three genes across six developmental stages of
seed were illustrated in Figure 8.
AhKR
and
AhENR
genes reached a maximum expression level at 25 DAP
and showed a downward trend thereafter. In contrast,
AhHD
gene expressed in an irregular course during
seed development and had high expression at 25 and
60 DAPs. These results indicated that these genes may
have different biochemical functions during vegetative
growth and seed development.