Basic HTML Version
Genomics and Applied Biology
,
2012, Vol.3 No.3, 1
-
7
http://gab.sophiapublisher.com
Table 1 Homology comparisons of the four differentially expressed gene fragments
Clone
Size (bp)
Accession No.
Homology
Identity (%)
Species
1 130
475
XM 002893420.1
GYF domain containing protein
87
Arabidopsis
1 252
337
D10840.1
5.8S, 25S, 18S rRNA fragment
97
Arabidopsis
1 301
391
BT021929.1
At1g62305
gene fragment
91
Arabidopsis
1 319
169
DQ658218.1
Polygalacturonase (
MF6
gene fragment)
100
Brassica
of
MF6
gene were suitable to use 2
-ΔΔCt
method for
quantitative expression (Figure 5). The expression
quantity of
MF6
gene in the buds of different fertility
and lengths were obtained after the quantitative expression
analysis by the 2
-ΔΔCt
method (Table 3). The expression
quantities of
MF6
gene in the male fertile buds were
apparently higher than in the male sterile buds during
the whole development period. The expression quantity
of
MF6
gene in the male fertile buds was 18.53 times
of that in the male sterile buds when the length of
buds was 1.00~1.50 mm, and 43.15 times when the
length of buds was 3.50~4.00 mm.
3
Table 2 Stability comparisons of reference genes
Genes
Coefficient variance
M value
TIP41
0.3431
0.6618
ACTIN
0.2946
0.7064
UBC21
0.1575
0.5284
Figure 4 The detection of amplification specificity and efficiency
of
UBC21
gene
Note: A: Melt curve; B: Standard curve
Figure 5 The detection of amplification specificity and efficiency
of
MF6
gene
Note: A: Melt curve; B: Standard curve
2 Discussions
Zhang et al (2008) obtained the differentially expressed
BcMF6
gene using the cDNA-AFLP technology and
the RACE technology in Chinese cabbage (
Brassica
rapa
L.). They constructed an antisense RNA expression
vector with the tapetum specific promoter A9, BcA9,
and transferred it into the cabbage recipient plants
though the
Agrobacterium
-mediated transformation. They
concluded that the
BcMF6
gene was pollen specifically
expressed
PG
(polygalact-uronase) gene during the
maturation process of tapetum and pollens after they
detected the morphology after the cytology and the
molecular markers were conducted in the transformed
Kan-resistant cabbage plants. In this study, we also
obtained a cDNA fragment which was 100% homologous
to the
PG
gene
MF
in the male fertile buds and the
male sterile buds induced with a male sterilizing agent
“Hua-sha-lingWP1”. The results of quantitative expression