2.2. Starch and chlorophyll analysis
Chlorophyll was extracted from 0.35 cm2 leaf discs in 80% acetone
and quantified as described by Lichtenthaler and Wellburn
[18]. Leaf samples were taken at the end of the day and night periods
and extracted repeatedly with acetone followed by water. Pellets
were boiled for 40 min with 1 ml of 0.1 M sodium acetate–0.1 M
acetic acid and then hydrolyzed overnight at 40 ◦C using 3 units
of -amylase and 0.25 units of amyloglucosidase. Concentration
of glucose was determined using an amplex-red glucose assay kit
(Molecular Probes, Inc.).
2.3. Evaluation of agronomic traits of rice
Transgenic rice seeds were germinated on sterilized water containing
30 mg L−1 hygromycin and grown in soil in a greenhouse.
For analysis of growth, WT and UpReg-1 overexpressing rice (T2
generation: UpReg1-16, 2-3 and 2-19) plants were grown under
natural environmental conditions in an experimental field plot at
the National Institute of Crop Science in Suwon, Korea. 30-day-old
rice seedlings were transplanted into six rows of 25 plants per row
at a planting distance of 30 cm
×
15 cm. Twenty plants from the
middle of the central row in each plot were used for collection of
trait data. The traits measured were: culm length (CL, cm), panicle
length (PL, cm), panicle number per hill (PNPH), spikelet number
2.2. Starch and chlorophyll analysisChlorophyll was extracted from 0.35 cm2 leaf discs in 80% acetoneand quantified as described by Lichtenthaler and Wellburn[18]. Leaf samples were taken at the end of the day and night periodsand extracted repeatedly with acetone followed by water. Pelletswere boiled for 40 min with 1 ml of 0.1 M sodium acetate–0.1 Macetic acid and then hydrolyzed overnight at 40 ◦C using 3 unitsof -amylase and 0.25 units of amyloglucosidase. Concentrationof glucose was determined using an amplex-red glucose assay kit(Molecular Probes, Inc.).2.3. Evaluation of agronomic traits of riceTransgenic rice seeds were germinated on sterilized water containing30 mg L−1 hygromycin and grown in soil in a greenhouse.For analysis of growth, WT and UpReg-1 overexpressing rice (T2generation: UpReg1-16, 2-3 and 2-19) plants were grown undernatural environmental conditions in an experimental field plot atthe National Institute of Crop Science in Suwon, Korea. 30-day-oldrice seedlings were transplanted into six rows of 25 plants per rowat a planting distance of 30 cm×15 cm. Twenty plants from themiddle of the central row in each plot were used for collection oftrait data. The traits measured were: culm length (CL, cm), paniclelength (PL, cm), panicle number per hill (PNPH), spikelet number
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