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Figure 1.

Characterization of ygl7 and ygl7-NIL phenotypes and leaf pigments.

A. Phenotypes of wild type (WT) and ygl7 at seedling, booting, and heading stages. B. Leaf-color comparison between WT and ygl7. C. Leaf pigment contents of WT and ygl7 at booting and heading stages. D. Phenotypes of ygl7-NIL and its parents, NPB (acceptor) and ygl7 (donor). E. Comparison of leaf pigments contents of ygl7-NIL with its parents, NPB and ygl7 at booting and heading stages. Values are the mean ± SD of three replicates.

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Table 1.

The main agronomic traits of ygl7's propagation.

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Table 2.

F1 agronomic traits.

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Table 3.

Segregation of F1 and F2 populations from three crosses.

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Figure 2.

Map-based cloning of the Ygl7 locus.

A. Preliminary map of the Ygl7 locus between RM1038 and SFP-3-6 based on analysis of 368 F2 (ygl7/NPB) individuals. Numbers below indicate the genetic distance (cM) between the markers. B. Fine mapping of the Ygl7 locus, which was narrowed down to a 64.8 kb genomic DNA region between RM10106 (2 recombinants) and STS3-1 (6 recombinants) based on 651 F2 individuals. C. Ygl7 locus was covered by two BAC contigs AC135595 and AC137507. D. There were 12 expressed genes identified in the fine mapping of Ygl7's area. E. One candidate Ygl7 gene (Os03g59640) was found to contain one base replacement (1883, T→C) resulting in one amino acid mutation (628, Leu→Ser). F. PCR analysis of the F2 yellowy-green leaf plants from Ygl7 crossed with NPB. Marker, RM16108; P1, Ygl7; P2, NPB; M, 100 bp low ladder; others are individual yellowy-green leaf plants of the F2 from Ygl7 crossed with NPB.

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Figure 3.

Expression analysis of Ygl7.

A. Leaf phenotypes in the complementation test. B. The PCR detection of HPT in some transgenic lines. CK1, H2O; CK2, normal plant of NPB; CK3, plasmid pLYL18; D16, the transgenic plant without the YGL7 gene; D5, D18, and D28 are the transgenic plants with the YGL7 gene. C. Pigment contents. Values are the mean ± SD of three replicates.

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Figure 4.

RNAi analysis of Ygl7.

A. and B. Phenotypes of leaf in the RNAi plants at 5 leaves. C. The expressions of ChlD (Ygl7) genes in tRNAi plants at 5 leaves. D. and E. Phenotypes of leaf in the RNAi plants at 7 leaves; old leaves were dead. F. The expressions of ChlD (Ygl7) genes in the RNAi plants at 7 leaves. Values are the mean ± SD of three replicates. Di1, Di2 and Di3 are the RNAi plants.

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Figure 5.

Real-time PCR expression analysis of genes associated with chlorophyll biosynthesis and photosynthesis in WT and ygl7.

Genes involved in chlorophyll biosynthesis included ChlD(YGL7), ChlI, ChlH, Hema1, PoRA, and Ygl1. Genes involved in photosynthesis included Cab1R, Cab2R, PsaA, PsbA, and RbcL. Values are the mean ± SD of three replicates.

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Figure 6.

Chlorophyll Fluorescence in three leaf-color mutants.

Fo: immobilized fluorescent, Fo coincides chlorophyll content; Fv'/Fm': effective photochemical quantum yield from PSII; ETR: the electron-transfer; qP: photochemistry quenching; qN: non-photochemistry quenching.

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Figure 7.

Characterization of the OsChlD's alleles.

A. Phenotypes of the three mutants and their crossed offspring. The chl1 mutant is the most yellow of the three. B. Pigment contents of leaves. Values are the mean ± SD of three replicates. The chl has the lowest amount of pigment. All F1 and F2 offspring's pigment amounts fall between the amounts contained in their parents.

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