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Understanding the genetic mechanism underlying folate biosynthesis and accumulation in rice would be beneifcial for breeding high folate content varieties as a cost-effective approach to addressing widespread folate deifciency in developing countries. In this study, the inheritance of rice grain folate content was investigated in the Lemont/Teqing recombinant inbred lines and the Koshihikari/Kasalath//Koshihikari backcross inbred lines. 264 F12 recombinant inbred lines (RILs) and 182 BC1F10 backcross inbred lines (BILs) with their parents planted in randomized complete blocks with two replicates in 2010, and RILs harvested in 2008 were used for QTL detection using inclusive composite interval mapping (ICIM) method. In the RIL population, two QTLs, denoted by qQTF-3-1and qQTF-3-2(QTF, quantitative total folate), explaining 7.8% and 11.1-15.8% of the folate content variation were detected in one or two years, respectively. In the BIL population, a QTL, denoted by qQTF-3-3,was detected, explaining 25.3% of the variation in folate content. All the positive alleles for higher folate content were from the high-folate parents, i.e., Teqing and Kasalath. The known putative folate biosynthesis genes do not underlie the QTLs detected in this study andtherefore may be novel loci affecting folate content in milled rice. QTLs identiifed in this study have potential value for marker assisted breeding for high-folate rice variety.