Purine nucleotide biosynthetic gene GARS controls early chloroplast development in rice (Oryza sativa L.).

KEY MESSAGE: GARS encodes an enzyme catalyzing the second step of purine nucleotide biosynthesis and plays an important role to maintain the development of chloroplasts in juvenile plants by affecting the expression of plastid-encoded genes. A series of rice white striped mutants were previously described. In this research, we characterized a novel gars mutant with white striped leaves at the seedling stage. By positional cloning, we identified the mutated gene, which encodes a glycinamide ribonucleotide synthetase (GARS) that catalyzes the second step of purine nucleotide biosynthesis. Thylakoid membranes were less abundant in the albinic sectors of mutant seedling leaves compared to the wild type. The expression levels of genes involved in chlorophyll synthesis and photosynthesis were changed. Contents of ATP, ADP, AMP, GTP and GDP, which are crucial for plant growth and development, were decreased in the mutant seedlings. Complementation and CrispR tests confirmed the role of the GARS allele, which was expressed in all rice tissues, especially in the leaves. GARS protein displayed a typical chloroplast location pattern in rice protoplasts. Our results indicated that GARS was involved in chloroplast development at early leaf development by affecting the expression of plastid-encoded genes.

Overexpression of OsbHLH107, a member of the basic helix-loop-helix transcription factor family, enhances grain size in rice (Oryza sativa L.).

BACKGROUND: Grain size, which is determined by grain length, grain width, and grain thickness, is an important determinant for grain yield in rice. Identification and characterization of new genes that are associated with grain size will be helpful for the improvement of grain yield in rice. RESULTS: We characterized the grain size mutant, larger grain size 1 (lgs1), derived from rice activation-tagged T-DNA insertion lines. Histological analysis showed that increased cell numbers in the longitudinal direction of spikelet hulls was responsible for the grain mutant phenotype in lgs1. Quantitative real-time PCR (qRT-PCR) analysis further showed that the expression levels of genes associated with the cell cycle in the young panicles of the lgs1 were higher than those in the wild type (WT), which might result in the increased cell numbers in lgs1 spikelet hulls. Insertion site analysis together with transgenic experiments confirmed that the lgs1 phenotype was caused by enhanced expression of truncated OsbHLH107, corresponding to the nucleotide (nt) 331-846 region (i.e., the transcriptional activation region of OsbHLH107) of the OsbHLH107 coding sequence (CDS). OsbHLH107 is a nucleus-localized bHLH transcription factor, which can form a homodimer with itself. Phylogenetic analysis showed that OsbHLH107 belonged to the same subfamily as OsPILs. OsPIL13 (OsPIL1) and OsPIL16 (APG) were reported to regulate grain size in rice. By transgenic experiments, we found that OsPIL11 could also regulate grain size. CONCLUSION: We concluded that OsbHLH107 and its homologs are important regulators of grain size development and might be useful for grain yield improvement in rice.

Fine Mapping and Candidate Gene Analysis of qSTL3, a Stigma Length-Conditioning Locus in Rice (Oryza sativa L.).

The efficiency of hybrid seed production can be improved by increasing the percentage of exserted stigma, which is closely related to the stigma length in rice. In the chromosome segment substitute line (CSSL) population derived from Nipponbare (recipient) and Kasalath (donor), a single CSSL (SSSL14) was found to show a longer stigma length than that of Nipponbare. The difference in stigma length between Nipponbare and SSSL14 was controlled by one locus (qSTL3). Using 7,917 individuals from the SSSL14/Nipponbare F2 population, the qSTL3 locus was delimited to a 19.8-kb region in the middle of the short arm of chromosome 3. Within the 19.8-kb chromosome region, three annotated genes (LOC_Os03g14850, LOC_Os03g14860 and LOC_Os03g14880) were found in the rice genome annotation database. According to gene sequence alignments in LOC_Os03g14850, a transition of G (Nipponbare) to A (Kasalath) was detected at the 474-bp site in CDS. The transition created a stop codon, leading to a deletion of 28 amino acids in the deduced peptide sequence in Kasalath. A T-DNA insertion mutant (05Z11CN28) of LOC_Os03g14850 showed a longer stigma length than that of wild type (Zhonghua 11), validating that LOC_Os03g14850 is the gene controlling stigma length. However, the Kasalath allele of LOC_Os03g14850 is unique because all of the alleles were the same as that of Nipponbare at the 474-bp site in the CDS of LOC_Os03g14850 among the investigated accessions with different stigma lengths. A gene-specific InDel marker LQ30 was developed for improving stigma length during rice hybrid breeding by marker-assisted selection.