Fig 1.
A-C, phenotype of IR64 and w67, 5-day-old (A), 9-week-old (B) and 14-week-old (C); D, Leaf blade of 4-week-old IR64 and w67; E, panicles of IR64 and w67.
Table 1.
The main agronomic traits of w67 and wild type IR64.
Table 2.
The pigment contents in w67 and wild-type IR64 (mg/g fresh weight).
Fig 2.
Chloroplast ultrastructure of IR64 and w67.
A-C, 4-week-old IR64; D-F, 4-week old w67; C chloroplast, M mitochondria, N nucleus, S starch granule, G granum, SL stroma lamella.
Table 3.
Chlorophyll fluorescence parameters in the 11-week-old w67 mutant and wild-type IR64.
Table 4.
Genetic analysis of the yellow-green phenotype in w67.
Fig 3.
A, The w67 locus was mapped to short arm of chromosome 3 between SSR markers RM14288 and RM14412; B, The w67 locus was narrowed down to about 274-kb region between RM6013 and RM569 using 298 F2 mutant individuals from cross of w67/R9308; C, The w67 locus was further mapped to about 160-kb region covering three BAC clone AP009053, AC098693 and AC140005 between InDel maker ID32 and RM569 using 503 F2 mutant type plants from cross of w67/Moroberekan; D, 26 ORFs were predicted in the mapped region and a single nucleotide substitution from A to T at position 130 in LOC_Os03g03990 was identified. Markers used for mapping are listed in S2 Table.
Fig 4.
Functional complementation at the w67 allele.
A, Phenotypes of w67, IR64 (WT) and complemented plants (pw67-1, pw67-2 and pw67-3); B, Nucleotides at the mutation sites in w67, IR64 and complemented plants; C, Levels of Chl a, Chl b and Cars in w67, IR64 and complemented plants at 8-week old. **, Highly significance at P ≤ 0.01.
Fig 5.
Domain organization of OscpSRP43 and structural details of ANK 1–3.
A, OscpSRP43 is composed of three CDs (red ellopses) and three ankyrin repeats (grey boxes); B, Structure of ankyrin-repeat domains (ANK1, residues 123–152; ANK2, 157–188; ANK3, 194–223). CD, chromodomain; ANK, anyrin-repeat domain.
Fig 6.
Sequence aligment and phylogenetic analysis of OscpSRP43 homologs.
A, Comparison of amino acid sequences of OscpSRP43 homologs. Residues identical to OscpSRP43 are shaded. Accession numbers for the respective protein sequences are as follows: Zea mays (NP_001168899); Sorghum bicolor (XP_002465920); Setaria italica (XP_004985832); Oryza sativa (NP_001048866); Brachypodium distachyon (XP_003562136); Glycine max (XP_003537753); Arabidopsis thaliana (NC_003071) and Chlamydomonas reinhardtii (AGC59877). B, Dendrogram of OscpSRP43 hommologs. The Neighbor-joining tree using percentage identities was constructed based on a multiple sequence alignment generated with the program MEGA 5.1.
Fig 7.
Expression of OscpSRP43 and subcellular localization of OscpSRP43.
A, Relative expression of OscpSRP43 in different organs of the wild type IR64 at the heading stage; B, Transient expression of GFP protein in rice protoplasts; C, Transient expression of OscpSRP43:GFP fusion protein in rice protoplasts; GFP, green fluorescence protein and OscpSRP43:GFP fusion protein; Auto, Chl auto fluorescence; Bright, bright field; Merged, merged image of GFP, Auto and Bright.
Fig 8.
Real-time quantitative reverse transcription PCR analysis.
The accession numbers and primer sequences of 16 genes associated with Chl biosynthesis and photosynthesis in IR64 and w67 are listed in S3 Table. The expression levels were determined by real-time PCR and normalized to the ubiqutin. The values are presented as the mean ± SD from three biological replicates. *, Significance at P ≤ 0.05; **, Highly significance at P ≤ 0.01.