Background: Effective tiller number (ETN) has a pivotal role in determination of rice (Oryza sativa L.) grain yield. ETN is a complex quantitative trait regulated by both genetic and environmental factors. Despite multiple tillering-related genes have been cloned previously, few of them have been utilized in practical breeding programs. Results: In this study, we conducted a genome-wide association study (GWAS) for ETN using a panel of 490 rice accessions derived from the 3 K rice genomes project. Thirty eight ETN-associated QTLs were identified, interestingly, four of which colocalized with the OsAAP1, DWL2, NAL1, and OsWRKY74 gene previously reported to be involved in rice tillering regulation. Haplotype (Hap) analysis revealed that Hap5 of OsAAP1, Hap3 and 6 of DWL2, Hap2 of NAL1, and Hap3 and 4 of OsWRKY74 are favorable alleles for ETN. Pyramiding favorable alleles of all these four genes had more enhancement in ETN than accessions harboring the favorable allele of only one gene. Moreover, we identified 25 novel candidate genes which might also affect ETN, and the positive association between expression levels of the OsPILS6b gene and ETN was validated by RT-qPCR. Furthermore, transcriptome analysis on data released on public database revealed that most ETN-associated genes showed a relatively high expression from 21 days after transplanting (DAT) to 49 DAT and decreased since then. This unique expression pattern of ETN-associated genes may contribute to the transition from vegetative to reproductive growth of tillers. Conclusions: Our results revealed that GWAS is a feasible way to mine ETN-associated genes. The candidate genes and favorable alleles identified in this study have the potential application value in rice molecular breeding for high ETN and grain yield.

Background: Grain size is a key agronomic trait that is directly associated with grain yield in rice. Although several genes related to grain size in rice have been identified, our understanding of the mechanism of grain development is still limited. Results: In this study, we reported the characterization of a novel seed size mutant mini seed 2 (mis2), in which the grain showed reduced length, width and thickness along with wrinkled surface. Microscopic analysis revealed that the spikelet epidermal cell size was reduced but the cell number was increased in the mis2 mutant, suggesting that MIS2 controls grain size by coordinately regulating epidermal cell size and cell number. Map-based cloning revealed that MIS2 encodes a receptor-like kinase CRINKLY4 (CR4) which showed the highest expression in developing panicles. The MIS2 protein is localized primarily on the plasma membrane along with the endosome. However, the Arg258Gln mutation located in extracellular domain in the mis2 mutant disturbed its subcellular localization. Additionally, three major haplotypes of MIS2 were identified in the japonica, indica and aus rice cultivars. The 18-bp InDel (insertion and deletion) in the 5′-UTR (untranslated region) caused different expression level of MIS2 in haplotypes. Conclusions: We reported a key role of OsCR4 in controlling grain size and shape by coordinately regulating epidermal cell size and cell number. The Arg258 in the extracellular seven-repeat domain is essential for the correct subcellular behavior and function of the OsCR4 protein.

Inflorescence architecture is important for rice (Oryza sativa) grain yield. The phytohormone cytokinin (CK) has been shown to regulate rice inflorescence development; however, the underlying mechanism mediated by CK perception is still unclear. Employing a forward genetic approach, we isolated an inactive variant of the CK receptor OHK4/OsHK4 gene named panicle length1 (pal1), which shows decreased panicle size due to reduced inflorescence meristem activity. A 2-amino acid deletion in the long α-helix stalk of the sensory module of OHK4 impairs the homodimerization and ligand-binding capacity of the receptor, even though the residues do not touch the ligand-binding domain or the dimerization interface. This deletion impairs CK signaling that occurs through the type-B response regulator OsRR21, which acts downstream of OHK4 in controlling inflorescence size. Meanwhile, we found that IDEAL PLANT ARCHITECTURE1(IPA1)/WEALTHY FARMER’S PANICLE (WFP), encoding a positive regulator of inflorescence meristem development, acts downstream of CK signaling and is directly activated by OsRR21. Additionally, we revealed that IPA1/WFP directly binds to the OHK4 promoter and upregulates its expression through interactions with two TCP transcription factors, forming a positive feedback circuit. Altogether, we identified the OHK4-OsRR21-IPA1 regulatory module, providing important insights into the role of CK signaling in regulating rice inflorescence architecture.

Direct seeding (DS) of rice gained much attention due to labor scarcity and unavailability of water. However, reduced emergence and poor seedling establishment are the main problems of DS which causes significant yield losses. Herein, DS-associated seedling traits of three major rice groups, i.e., Indica (Ind), Japonica (Jap), and aus-type (Aus), were evaluated by using an improved mesocotyl elongation assay. The associations among different traits at the seedling and maturity stage were also studied. Significant variation was observed among the cultivars of different rice groups. The Aus group cultivars showed higher mean values for coleoptile (C, 3.85 cm), mesocotyl (MC, 4.17 cm), shoot length (SL, 13.64 cm), panicle length (PL, 23.44 cm), tillers number (T, 15.95), culm length (CL, 105.29 cm), and plant height (PH, 128.73 cm), while the Indica and Japonica groups showed higher mean values of grain length (GL, 8.69 mm), grain length/width ratio (GL/WR, 3.07), and grain width (GW, 3.31 mm), with 1000 grain weight (TGWt, 25.53 g), respectively. Pairwise correlation analysis showed that MC, C, and SL were positively correlated among themselves and with PL, CL, and PH. Moreover, based on principal component analysis (PCA), C, MC, SL, CL, and PH were identified as the major discriminative factors in the rice cultivars. This study describes the development of desired DS rice variety with long MC and semidwarf in height and suggests that Aus group cultivars can be used as the donor parents of favorable DS-associated traits in rice breeding programs.

Pea root rot is a major soil-borne disease that threatens global pea production. In this study, diseased pea root samples were collected from Zhejiang Province for pathogen isolation and identification. The mycelial growth rate method was used to evaluate the sensitivity of the pathogens to different fungicides, and transcriptome analysis was performed to investigate the response mechanisms underlying pea root responses to pathogen infection. A total of eight fungal pathogens were identified, including Fusarium solani, F. commune, F. oxysporum, F. proliferatum, F. tricinctum, F. andiyazi, Alternaria alternata, and Didymella pinodella. Among them, F. solani and F. commune were the dominant pathogens. Both pathogens showed high sensitivity to prochloraz, with EC₅₀ values of 0.051 and 0.030 mg·L⁻¹, respectively. Transcriptome sequencing revealed that peas respond to pathogen infection through pathways related to secondary metabolism and plant hormone signal transduction. Furthermore, 14 candidate genes potentially involved in resistance to pea root rot were screened.