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template and PCR products as the probe. The PCR
products was purified and labeled with DIG High
Prime DNA Labeling and Detection Starter Kit
Ⅰ
(Roche) and hybridization, and washing conditions
were performed as follows: prehybridized with 10 mL
of DIF Easy Hyb solution at 42
℃
for 30 min. The
labeled probe (25 ng/mL) was denatured at 95~100
℃
for 5 min and put on ice immediately. And then
translated the probe into the heated hybridization
solution and mixed. Discarding the prehybridization
solution and adding the hybridization solution with
probe and hybridized at 45
℃
overnight. The
membranes were washed once at 15~25
℃
in 2×SSC
with 0.1% SDS for 10 min and twice at 65~68
℃
for
30 min in 0.5×SSC buffer with 0.1% (w/v) SDS. At
last, the hybridization signal was detected with
scanner (Typhoon Trio).
3.5 Recording the flowering time
The time that the heart leaves began to change color
after ethylene treatment was recorded for 5 plants for
each repeat. The average value was used for comparing.
Authors
'
Contributions
ZYL and LX were responsible for experimental design and the
experiment direction; ZLY carried out most of the work; HQC
took part in the work; ZYL and LX were responsible for data
analysis, paper writing and modification. All of them had read
the final version of this paper and agreed with the authors’ credits.
Acknowledgements
This work was supported in part by Natural Science Foundation
of Hainan Province Grant 808191 and The Basic Scientific
Research Funds Operating Expenses for Central Level Institutes
Grant PZS004.
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