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3.2 Extraction and detection of water-soluble
oligosaccharides
Whole wheat flour (0.1000 g) was accurately weighed
before extraction of water-soluble oligosaccharides in
5 ml double-distilled water for 10 h at 4°C. The
samples were then subjected to ultrasonic extraction
for 30 min and centrifuged at 4 500 r/min for 30 min.
Supernatants were filtered with C18
-
SPE and 0.45
µm
filter paper.
Water-soluble saccharide content was determined by
high-pressure liquid chromatography (HPLC) with a
Waters 515 pump and refractive index detector
(Waters 2410, USA) (Zhu and Yang, 2005). Com
pounds were separated on a 4.6×250 mm Inertsil NH2
column (Dikma, Japan) with the mobile phase
consisting of 25% water and 75% acetonitrile (flow
rate, 1 mL/min). The column temperature was 40°C.
3.3 QTL analysis
The water-soluble saccharide content data and the
linkage map of the Sh Ch population, which were
reported previously (Li et al., 2007), were used for
QTL analysis. This map included 381 loci on all the
wheat chromosomes, comprising 167 SSR, 94
EST-SSR, 76 ISSR, 26 SRAP, 15 TRAP, and 3 Glu
loci, and covering 3 636.7 cM with an average distance
of 14.8 cM between markers. QTL mapping was
conducted using QTLMapper 1.6 based on a mixed-
model (Wang et al., 1999). The walking speed chosen
for all QTLs was 1.0 cM. Additive effects of detected
QTLs were estimated by Bayesian test. A QTL was
considered significant at a LOD peak value 2.5 (Zhao
et al., 2009).
Authors’ contributions
XYF conceived the overall study, performed the experiment designs, carried
out the trait phenotyping, and drafted the manuscript. ZLQ took part to the
data analysis and the writing.SSL obtained and analyzed the QTL data and
was involved in the writing. All authors read and approved the final
manuscript.
Acknowledgements
We thank Shubo Gu (State Key Laboratory of Crop Biology
，
Agronomy
College of Shandong Agriculture University, Tai
’
an, China) for help and
advice on the experiment.
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