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Genomics and Applied Biology
,
2012, Vol.3 No.3, 1
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Table 3 The expression quantity of
MF6
gene in the buds with different lengths and different fertilities
Male sterile bud
Male fertile bud
Bud length
Expression
quantity
Standard
deviation
Bud length
Expression
quantity
Standard
deviation
Times
﹡
A1 (0.00~1.00 mm)
3.71
0.15
B1 (0.00 ~1.00 mm)
4.16
4.91
1.12
A2 (1.00~1.50 mm)
5.19
1.53
B2 (1.00 ~1.50 mm)
96.11 14.37
18.53
A3 (1.50~2.00 mm)
1.74
0.15
B3 (1.50 ~2.00 mm)
9.99
3.98
5.75
A4 (2.00~2.50 mm)
1.00
0.35
B4 (2.00 ~2.50 mm)
5.05
1.10
5.05
A5 (2.50~3.00 mm)
2.57
0.41
B5 (2.50 ~3.00 mm)
9.69
2.69
3.77
A6 (3.00~3.50 mm)
21.81
1.92
B6 (3.00 ~3.50 mm)
139.87
34.91
6.41
A7 (3.50~4.00 mm)
92.15
8.85
B7 (3.50 ~4.00 mm) 3 976.70
444.48
43.15
Note:
﹡
Ratio of the amount of
MF6
gene expression in the male fertile flower buds over that in male sterile flower buds
showed that this gene was highly associated with the
fertility performance and bud development in rapeseed
(
Brassica napus
L.). However, when the bud lengths
were more than 3.5 mm, and the development of
pollens was completed, the expression of
MF6
gene
was still remained remarkably higher in the male
fertile buds than that of the male sterile buds. Therefore,
the expression of
MF6
gene was not only related to
the development of pollens, but might also be involved
in the growth and development of other floral organs.
According to the results in 1.1, the male sterile flowers
had not only empty anthers but also shorter filaments.
So
MF6
gene may also be associated with the elongation
of filaments in
Brassica napus
L.
Tang et al (2009) observed that the development of
pollens was obviously differentiated in the male sterile
buds during the period of pollen mother cell stage and
the tetrad spore stage in R121 induced by "Hua-sha-
ling WP1". In this study, the results showed that the
expression of
MF6
gene was
obviously
inhibited in
the male sterile buds when the bud length was
1.00~1.50 mm. According to the observation of Gong
(2008) on the relationship between bud length and
developmental stage of pollens in rapeseed, the
development of pollen was at meiosis stage when
flower bud was about 1.5 mm in length. This was also
an important development stage of tapetum in the
rapeseed flower buds. Therefore, the later meiosis
stage of pollen mother cells was an important stage for
application of "Hua-sha-ling WP1" to induce pollen
abortion and produce male sterility in R121. The
important period for the application of "Hua-sha-ling
WP1" to produce male sterility in R121 could also provide
a strong scientific basis and a technical guidance for
studies and utilization of chemical hybridizing agents
to induce male sterility in
Brassica napus
L.
3 Materials and Methods
3.1 Materials and reagents
The experimental material R121 is a stable pure
B. napus
line with normal male fertility, which was provided by
the Leshan Academy of Agricultural Sciences, Sichuan
Province, China. The male sterilizing agent "Hua-sha-
ling WP1" was provided by Fu Yunlong, Mianxian
Seed Administration Station, Shanxi Province. The
plasmids containing ACTIN, TIP41, UBC21 were
supplied by the present laboratory.
Trizol Reagent was purchased from the Invitrogen
Company. PCR select
TM
and cDNA substraction kit
were purchased from the Clontech Company. P
MD19-T
vector, DNA marker, reverse
Taq
polymerase,Trans-
criptase M-MLV, primescript RT reagent kit enzymes
(perfect real time), SYBR premix Ex
Taq
Ⅱ
(perfect
real time) were purchased from TaKaRa Company.
Plasmid Miniprep kit was purchased from TianGen
Company. AxyPrep PCR cleaning kit was purchased from
Axygen. Dig High Prime DNA Labeling and Detection
Starter Kit
Ⅰ
were purchased from Roche Company.
3.2 Treatments to the materials
On 20
th
October, 2009, the plant material R121 was
planted in 5 rows and 12 hills per row, with 25 cm hill
distance on the teaching and research farm of Sichuan
Agricultural University. Attention was paid to the control
of soil moisture and pest incidence after emergence.
Two plants were saved per hill and the extra plants
were removed after 2 months. The male sterilizing
chemical "Hua-sha-ling WP1" was applied at the initial
bolting stage with a concentration of 7 mg/mL. Clean
water was sprayed for the first two rows as a control
group, and the last two rows were sprayed with the
male sterilizing chemical solution, and the central row
was used as a spacer row. The spayed amount of
solution or water was 9 mL per plant.