Motoyuki Ashikari’s research while affiliated with Nagoya University and other places

In rice, internode elongation is a critical aspect of plant development and agricultural productivity. Previous morphological and histochemical studies using [³H]thymidine have visualized the cell division zone (intercalary meristem) in internodes. However, it has remained unclear how the intercalary meristem forms during stem development. In addition, while a pith cavity forms in the central part of the rice stem, the spatiotemporal relationship between pith cavity formation and intercalary meristem development is not well understood. Therefore, we performed histological analysis of intercalary meristem and pith cavity development using C9285, a deepwater rice variety that shows internode elongation from the vegetative growth stage. We classified the developmental stages of the stem into four stages based on the analysis of pith cavity formation using Trypan blue, Calcein-AM, and MitoRed staining, and visualized dividing cells using the Click-iT EdU imaging assay. In Stage 1, no pith cavity was formed. Vertical cell rows were observed between above the axillary bud attachment and the upper node, suggesting anticlinal divisions that lead to internode formation in the early stage of stem development. In Stage 2, the first pith cavity formed in the pith of the foot, which is the region of axillary bud attachment. Compared to cell division in the internode, that in the foot was significantly activated resulting in slight elongation from Stage 1 to Stage 2. In Stage 3, cell division in the foot ceased, while active cell division at the base of the internode led to significant vertical elongation. The second pith cavity formed due to cell death in the pith of the internode. In Stage 4, the two pith cavities connected to form a single large pith cavity. Although the intercalary meristem maintained cell division activity, the number of cell divisions decreased. Based on these results, we propose a model for stem development that involves two phases of elongation regulation: primary elongation involving slight elongation in the foot, and secondary elongation involving significant internode elongation due to the activation of cell division and cell elongation in the intercalary meristem. This is the first study to anatomically elucidate the spatiotemporal relationship between intercalary meristem development and pith cavity formation in rice stem development. It provides new insights for future research on rice stem development and studies of other grass species.

Historical and recent insights into the molecular mechanisms of escape and quiescence strategies employed by rice to survive flooding.

Stem length is a crucial agronomic trait in rice breeding. The short stature of rice dwarf mutants is caused by shortening of internodes, resulting in five distinct internode elongation patterns: dn, dm, d6, nl and sh. Several genetic studies have been conducted; however, the genetic mechanisms underlying these internode elongation patterns remain unclear. In this study, we examined two Daikoku dwarf (d1) mutants, T65(d1-1) and Kin(d1-7), which display contrasting internode elongation phenotypes. Anatomical observation revealed that T65(d1-1) exhibits a dm-type internode elongation pattern due to the lack of the second internode counted from the top, while Kin(d1-7) shows a dn-type pattern with a relatively elongated second internode. To identify the genetic factors influencing these phenotypes, we conducted a quantitative trait locus (QTL) analysis using two F2 populations derived from reciprocal crosses between them. The QTL analysis showed that the second internode length is regulated by three QTLs on chromosomes 4, 5, and 6. Epistatic effects were ob‍served through the analysis of F3 progenies, indicating that the combination of Kin(d1-7) alleles at these QTLs is associated with an increased second internode length. Furthermore, specific combinations of alleles result in varying degrees of elongation in the second internode, significantly impacting the internode elon­gation pattern. These findings contribute to a deeper understanding of the genetic factors influencing the internode elongation patterns in rice.

In rice, internode elongation is a critical aspect of plant development and agricultural productivity. Previous morphological and histochemical studies using [ ³ H]thymidine have visualized the cell division zone (intercalary meristem) in internodes. However, it has remained unclear how the intercalary meristem forms during stem development. In addition, while a pith cavity forms in the central part of the rice stem, the spatiotemporal relationship between pith cavity formation and intercalary meristem development is not well understood. Therefore, we performed histological analysis of intercalary meristem and pith cavity development using C9285, a rice variety that shows internode elongation from the vegetative growth stage. We classified the developmental stages of the stem into four stages based on analysis of pith cavity formation using trypan blue staining and visualized dividing cells by Click-iT EdU imaging assay. In Stage 1, no pith cavity was formed. Vertical cell rows were observed between the upper end of the axillary bud attachment and the upper node, suggesting anticlinal divisions that lead to internode formation in the early stage of stem development. In Stage 2, the first pith cavity formed in the pith of the foot, which is the region of axillary bud attachment. Compared to cell division in the internode, that in the foot was significantly activated resulting in slight elongation from Stage 1 to Stage 2. In Stage 3, cell division in the foot ceased, while active cell division at the base of the internode led to significant vertical elongation. The second pith cavity formed due to cell death in the pith of the internode. In Stage 4, the two pith cavities connected to form a single large pith cavity. Although the intercalary meristem maintained cell division activity, the number of cell divisions decreased. Based on these results, we propose a model for stem development that involves two phases of elongation regulation: primary elongation involving slight elongation in the foot, and secondary elongation involving significant internode elongation due to the activation of cell division and cell elongation in the intercalary meristem. This is the first study to anatomically elucidate the spatiotemporal relationship between intercalary meristem development and pith cavity formation in rice stem development. It provides new insights for future research on rice stem development and studies of other grass species.

Rice (Oryza sativa L.) and many other wetland plants form an apoplastic barrier in the outer parts of the roots to restrict radial O2 loss to the rhizosphere during soil flooding. This barrier facilitates longitudinal internal O2 diffusion via gas-filled tissues from shoot to root apices, enabling root growth in anoxic soils. We tested the hypothesis that Leaf Gas Film 1 (LGF1), which influences leaf hydrophobicity in rice, plays a crucial role in tight outer apoplastic barriers formation in rice roots. We examined the roots of a rice mutant (dripping wet leaf 7, drp7) lacking functional LGF1, its wild type, and an LGF1 overexpression line for their capacity to develop outer apoplastic barriers that restrict radial O2 loss. We quantified the chemical composition of the outer part of the root and measured radial O2 diffusion from intact roots. The drp7 mutant exhibited a weak barrier to radial O2 loss compared to the wild type. However, introducing functional LGF1 into the mutant fully restored tight barrier function. The formation of a tight barrier to radial O2 loss was associated with increased glycerol ester levels in exodermal cells, rather than differences in total root suberization or lignification. These results demonstrate that, in addition to its role in leaf hydrophobicity regulation, LGF1 plays an important role in controlling the function of the outer apoplastic barriers in roots. Our study suggests that increased deposition of glycerol esters in the suberized root exodermis establishes a tight barrier to radial O2 loss in rice roots.

The awn is a bristle-like appendage that protrudes from the seed tip and plays a critical role in preventing feed damage and spreading habitats in many grass species, including rice. While all wild species in the Oryza genus have awns, this trait has been eliminated in domesticated species due to its obstructive nature to agricultural processes. To date, several genes involved in awn development have been identified in wild rice, Oryza rufipogon and Oryza barthii which are ancestral species of cultivated rice in Asia and Africa, respectively. However, the responsible genes for awn development have not been identified in other wild rice species even though multiple QTLs have been reported previously. In this study, we identified An7 gene re‍sponsible for awn development in two wild rice species, Oryza glumaepatula and Oryza meridionalis. An7 encodes a cytochrome P450 enzyme and is homologous to D2/CYP90D2, a known brassinosteroid biosynthesis enzyme in rice. The identification of An7 provides insight into a distinct molecular mechanism underlying awn development that occurs in geographically separated environments.

To counteract the growing population and climate changes, resilient varieties adapted to regional environmental changes are required. Landraces are valuable genetic resources for achieving this goal. Recent ad‍vances in sequencing technology have enabled national seed/gene banks to share genomic and genetic in‍formation from their collections including landraces, promoting the more efficient utilization of germplasms. In this study, we developed genomic and genetic resources for Myanmar rice germplasms. First, we assembled a diversity panel consisting of 250 accessions representing the genetic diversity of Myanmar indica varieties, including an elite lowland variety, Inn Ma Yebaw (IMY). Our population genetic analyses illustrated that the diversity panel represented Myanmar indica varieties well without any apparent population structure. Second, de novo genome assembly of IMY was conducted. The IMY assembly was constructed by anchoring 2888 contigs, which were assembled from 30× coverage of long reads, into 12 chromosomes. Although many gaps existed in the IMY genome assembly, our quality assessments indicated high completeness in the gene-coding regions, identical to other near-gap-free assemblies. Together with dense variant information, the diversity panel and IMY genome assembly will facilitate deeper genetic research and breeding projects that utilize the untapped Myanmar rice germplasms.

Paw San Hmwe (PSH) is an indigenous rice variety from Myanmar with a good taste, a pleasant fragrance, and excellent elongation ability during cooking. However, its low yield potential and strong photoperiod sensitivity reduce its productivity, and it is vulnerable to climate changes during growth. To improve the photoperiod insensitivity, yield, and plant stature of PSH, the high-yield genes Grain number 1a (Gn1a) and Wealthy Farmer’s Panicle (WFP), together with the photoperiod insensitivity trait, were introgressed into PSH via marker-assisted backcross breeding and phenotype selection. For the photoperiod insensitivity trait, phenotypic selection was performed under long-day conditions during the dry season. After foreground selection of Gn1a and WFP via simple sequence repeat genotyping, genotyping-by-sequencing was conducted to validate the introgression of target genes and determine the recurrent parent genome recovery of the selected lines. The improved lines were insensitive to photoperiod, and the Gn1a and WFP introgression lines showed significantly higher numbers of primary panicle branches and spikelets per panicle than the recurrent parent, with comparative similarity in cooking and eating qualities. This study successfully improved PSH by decreasing its photoperiod sensitivity and introducing high-yield genes via marker-assisted selection. The developed lines can be used for crop rotation and double-season cropping of better-quality rice.