| Soybean isoflavone is a kind of secondary metabolite of soybean,which is similar to estrogen.It not only has a good effect on the treatment and prevention of cance,but also can improve menopausal symptoms and delay female aging;also has good effect on prevention and treatment of osteoporosis,hyperlipidemia,cardiovascular disease,heart disease and other dementia elderly.In the field of medical and food health it has aroused great concern.How to improve the isoflavone content in soybean seeds and study its molecular mechanism have become an important goal of soybean quality breeding.The extraction methods of soybean isoflavones content was optimized and a set of standard of the laboratory extraction isoflavone with high performance liquid chromatography(HPLC)method was established in the experiment.Jinong 17 and Jining 18 soybean were selected to hybrid,and then established the population of recombinant inred line F2: 6 families as test materials.The total amount of soybean isoflavones and daidzin,soybeoside,Glycosides and other components were identified.The technique of SSR molecular marker was used to construct the genetic linkage map,and the QTLs related to isoflavone content were finally located.The main conclusions are as follows:1.Optimization of extraction of soybean isoflavones content of the method:Degreased soybean sample powder 250 mg,adding the volume of 80% of the chromatographic grade methanol 10 mL,placed at room temperature for 2 hours,after placing the ultrasonic power 200 W 80 ? water bath for 12 hours,12000 rpm centrifugation for 15 minutes,take the supernatant by 0.45 ?m microporous membrane filtration,soy isoflavones fluid to be stored under 4 ?.2.Optimization of soybean isoflavone content by HPLC:The mobile phase consisted of methanol-water(volume ratio 30:70).The column temperature was 40 ?,wavelength: 254 nm,flow rate: 1mL / min,injection volume: 10?L,injection time: 26 min,Using the peak area for the calculation method..3.Correlation analysis of isoflavones content in RIL population:Daidzin content and genistin content accounted for more than 90% of the total content of soybean isoflavones.The relationship between the size of each component: daidzin> genistin> glycitin.The correlation coefficients of total isoflavones and other ingredients were: genistin> daidzin> protein> fat> glycitin;The content of total isoflavones,daidzein and genistein was negative correlative with fat content,and was positive correlative with protein content.4.The QTL analysis of soybean isoflavones content in RIL population:A genetic linkage map covering 14 chromosomes was constructed using QTL IciMapping 4.0 software by RIL group which was established with Jinong 17 × Jinong 18 combination and SSR molecular marker technology.Its total length was 1365.73 cM and the average genetic distance was 17.51 cM.The SSR markers arrangement order in the linkage group is almost same with the public map.The results showed that 22 QTLs were related to the total amount of soybean isoflavones and the content of each component were detected,including QTLs related to daidzin content,QTLs related to genistin content,QTLs related to the content of daidzein,4 QTLs related to total isoflavones.The QTLs of isoflavone content located in this study were compared with those of QTLs in SoyBase library,according to the common genetic map of soybean in 2003(GmComposite 2003).The results indicated that 15 QTLs is the same as in the SoyBase library and there were 7 new QTLs,.Among them a QTL were related to the content of daidzin: qDaidzin-1-1;two QTLs were related to the content of the glycosides : QGlycitin-9-1,qGlycitin-12-1;four QTLs were related to genistin content: qGenistin-7-1,qGenistin-8-1,qGenistin-10-1,qGenistin-14-2. |
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