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Genomics and Applied Biology, 2011, Vol.2 No.5
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designing was based on the public sequence of
Nipponbare (http://rgp.dna.affrc.go.jp). Since sd1 confers
GA20ox -2 and shares homology with GA20ox -1
(Os03g0856700), the primer was designed from the
region showing low homology between GA20ox -2
and GA20ox -1, allowing distinct labeling of GA20ox -2
and GA20ox -1, respectively. PCR amplification took
place in a 20 µL containing: 10 ng genomic DNA, 1 µM
of each primer, 0.4 mM dNTPs, 1× GC Buffer Ⅰ, 2.5 mM
MgCl2, and 0.5 unit of LA Taq polymerase (Takara) in
a total volume of 20 µL. and using following cycling
conditions: 35 cycles, 94℃ 30 s, 58℃ 30 s, and
72℃ 1 min. PCR products were treated with the
restriction enzyme PmaCI (CAC↓GTG), which can
detect a single base substitution site of sd1 in the 1st
exon of sd1 from Hikarishinseiki according to Tomita
(2009). The transcription product of sd1 was then
analyzed.
3.2 Performance test of sd1 in the isogenic background
Performance tests were carried out in a paddy field at
Tottori University, Koyama, during 2006 and 2007.
‘Hikarishinseiki’ and ‘Koshihikari’ were sown on 20
April 2006 and on 21 April 2007, and 128 seedlings
per plot were transplanted with two replications on 15
and 17 May, respectively. Similar performance tests
were conducted at experimental stations in Miyagi,
Ibaraki, Kanagawa, Nagano, Mie, Toyama, Ishikawa
Kyoto, Wakayama, Hyogo, Shimane, Okayama, Toku-
shima, Ehime, Oita and Kumamoto, which are spread
across Japan.
The date was recorded as the heading date once when
50% of all panicles had emerged from the flag leaf
sheath. Days-to-heading was the number of days from
sowing date to heading date. Culm length, panicle
length, number of panicles, leaf length, and leaf width
were measured on 10 randomly selected individuals in
each plot. Thousand-grain weight, grain yield of brown
rice, grain quality, and eating quality were measured
on bulks of 50 individuals. The means of traits were
statistically compared using the t-test.
Author’s contributions
MT conceived and designed the study and wrote the manuscript.
MT and SM performed the experiments and analyzed the data.
Both of authors read and approved the final manuscript.
Acknowledgements
The authors would like to express our gratitude to Japan
Science and Technology Agency (JST) for the Grant-in-Aid for
Adaptable and Seamless Technology Transfer Program through
Target-driven R & D (No.08150094 and No.08001167) that
supported this work to Motonori Tomita. We thank to all those
who provided test data from prefectures across Japan.
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