CYP78A, a cytochrome P450 subfamily that includes rice (Oryza sativa L.) BIG GRAIN2 (BG2, CYP78A13) and Arabidopsis thaliana KLUH (KLU, CYP78A5), generate an unknown mobile growth signal (referred to as a CYP78A-derived signal) that increases grain (seed) size. However, the mechanism by which the CYP78A pathway increases grain size remains elusive. Here, we characterized a rice small grain mutant, small grain4 (smg4), with smaller grains than its wild type due to restricted cell expansion and cell proliferation in spikelet hulls. SMG4 encodes a multidrug and toxic compound extrusion (MATE) transporter. Loss of function of SMG4 causes smaller grains while overexpressing SMG4 results in larger grains. SMG4 is mainly localized to endoplasmic reticulum (ER) exit sites (ERESs) and partially localized to the ER and Golgi. Biochemically, SMG4 interacts with coat protein complex Ⅱ (COPⅡ) components (Sar1, Sec23, and Sec24) and CYP78As (BG2, GRAIN LENGTH 3.2 [GL3.2], and BG2-LIKE 1 [BG2L1]). Genetically, SMG4 acts, at least in part, in a common pathway with Sar1 and CYP78As to regulate grain size. In summary, our findings reveal a CYP78As–SMG4–COPⅡ regulatory pathway for grain size in rice, thus providing new insights into the molecular and genetic regulatory mechanism of grain size.

Background: Chloroplast development is coordinately regulated by plastid- and nuclear-encoding genes. Although many regulators have been reported to be involved in chloroplast development, new factors remain to be identified, given the complexity of this process. Results: In this study, we characterized a rice mutant lethal albinic seedling 1(las1)form of a 4-hydroxy-3-methylbut-2-enyl diphosphate reductase (OsHMBPP) that was targeted to the chloroplasts. The LAS1 mutation caused the albino lethal phenotype in seedlings. Transmission electron microscopy indicated that las1 were defective in early chloroplast development. LAS1 is preferentially expressed in leaves, implying its role in controlling chloroplast development. The expression levels of many chloroplast-encoded genes were altered significantly in las1. The expression levels of nuclear-encoded gene involved in Chl biosynthesis were also decreased in las1. We further investigated plastidic RNA editing in las1 and found that the edit efficiency of four chloroplast genes were markly altered. Compared with WT, las1 exhibited defective in biogenesis of chloroplast ribosomes. Conclusions: Our results show that LAS1/OsHMBPP plays an essential role in the early chloroplast development in rice.

Background: Plant plastidic caseinolytic protease (Clp) is a central part of the plastid protease network and consists of multiple subunits. The molecular functions of many Clps in plants, especially in crops, are not well known. Results: In this study, we identified an albino lethal mutant al3 in rice, which produces albino leaves and dies at the seedling stage. Molecular cloning revealed that AL3 encodes a plastid caseinolytic protease, OsClpR1, homologous to Arabidopsis ClpR1 and is targeted to the chloroplast. Compared with the wild type, chloroplast structure in the al3 mutant was poorly developed. OsClpR1 was constitutively expressed in all rice tissues, especially in young leaves. The OsClpR1 mutation affected the transcript levels of chlorophyll biosynthesis and chloroplast development-related genes. The RNA editing efficiency of three chloroplast genes (rpl2, ndhB, ndhA) was remarkably reduced in al3. Using a yeast two-hybrid screen, we found that OsClpR1 interacted with OsClpP4, OsClpP5, OsClpP2, and OsClpS1. Conclusions: Collectively, our results provide novel insights into the function of Clps in rice.

Grain size is one of the major factors determining rice grain yield. Nevertheless, our knowledge of the molecular mechanisms underlying the control rice grain size remains limited. Trihelix proteins are plant-specific transcription factors that regulate plant growth and development. However, their roles in modulating grain size in cereal crops are largely unknown. Here, we report the rice trihelix family gene Oryza sativa trihelix transcription factor related to grain size 1 (OsTGS1) as a novel regulator of grain size and weight. Mutation of OsTGS1 leads to large and heavy grains, whereas overexpression of OsTGS1 results in small and light grains. OsTGS1 regulates grain size by influencing cell division and cell expansion in spikelet hulls. OsTGS1 is expressed in various tissues, and its expression level increases during panicle development. The OsTGS1 protein is localized to the nucleus and exhibits transcriptional repressor activity. The screening of interacting proteins via a yeast two-hybrid assay revealed that OsTGS1 interacted with GSK3/SHAGGY-LIKE KINASE2 (GSK2), an important regulator of various agronomic traits, including grain size, in rice. Moreover, ostgs1 mutants are hypersensitive to exogenous brassinosteroid treatment, indicating that OsTGS1 may be involved in brassinosteroid signaling. Our study reveals the role of OsTGS1 in controlling grain size and provides a new gene resource for improving grain weight in rice.