Dominant complementary interaction between OsC1 and two tightly linked genes, Rb1 and Rb2, controls the purple leaf sheath in rice.

Two tightly linked genes for rice purple leaf sheath were identified via map-based cloning. Further analysis indicated that these two genes together with OsC1 co-regulating the purple leaf sheath. The purple color of the leaf sheath in rice is dependent on the accumulation of anthocyanins such as cyanidin 3-O-glucoside (C3G) and peonidin 3-O-glucoside (P3G). Although many genes related to leaf sheath color have been mapped, the genetic basis for leaf sheath color is not yet clear. Here, PSH1 (purple leaf sheath 1) was mapped to chromosome 1 using an F and a RIL population. Map-based cloning and transformation assays further divided PSH1 as two tightly linked bHLH genes, Rb1 and Rb2. Ectopic expression of these two genes resulted in substantial accumulation of C3G and P3G in the leaf blade, leaf sheath and pericarp. Single gene mutants displayed a faded purple leaf sheath or green leaf sheath in the top half of the leaf sheath, but double mutants displayed a green leaf sheath, indicating that both genes have dosage effects on anthocyanin synthesis. However, overexpression of Rb1 and Rb2 sharply decreased grain filling. A segregation ratio of green to purple was 15:1 observed in the F population from parents Minghui 63 and Xizang 2, which both had green leaf sheaths; these results demonstrate that dominant complementary interaction between OsC1 and Rb (Rb1 and Rb2) controls the purple leaf sheath. These findings systematically uncovered the genetic basis of leaf sheath color and provided alternative genes for breeding anthocyanin-rich rice.