Search bioRxiv⌕ Search

bioRxiv · 10.1101/2025.03.31.646432

Extremely Early Flowering and Large Grain Isogenic Japonica Rice Koshihikari Integrated with Gene e1 and GW2

Abstract

The extremely early flowering/large-grain isogenic Koshihikari was developed by combining the large-grain allele, GW2, derived from Inochinoichi with the year-round flowering allele, e1, from Kanto 79. We conducted four back crosses with Koshihikari as a recurrent parent by using an extremely early flowering/large-grain e1GW2 homozygote as a non-recurrent parent, which was segregated in B1F2 between Kanto 79 and a large-grain fixed F3 plant in Koshihikari x Inochinoichi. In the BC4F2 population, the e1GW2 homozygous phenotype was selected and fixed in BC4F3 as Kshihikari e1GW2. plants were segregated according to a ratio of 1 extremely early flowering/large-grain e1GW2 : 3 extremely early maturing/small-grain e1gw2 : 3 medium-maturing/large-grain e1GW2 : 9 medium-flowering/small-grain E1gw2 and e1GW2 homozygous isogenic Koshihikari [Koshihiakri e1GW2] was fixed in BC4F3. Whole genome sequencing of Koshihiakri e1GW2 proved that already known 1 deletion in GW2 at 8,147,416 bp on chromosome 2, and an e1 isogenic-specific SNP at 9,090,618 bp, which was 35,213 bp downstream to the 3{square} side of Ghd7 on chromosome 7. RT-qPCR analyses showed that the transcription of Ghd7 was suppressed in Koshihikari e1GW2 than in Koshihikari. The Koshihikari e1GW2 flowered 14 days earlier than Koshihikari and thousand grain weight of Koshihikari e1GW2 (27.8 g) was 18% larger than Koshihikari (23.6 g). We successfully integrated GW2 with e1 for the first time, especially in the genome of a globally produced Japonica leading cultivar Koshihikari. The Koshihikari e1GW2 was registered under plant varietal protection.

Explore related subjects

Keep this discovery

Explore connections, maps & timelines

BibTeXRIS

Tomita, M., Arai, K.. 2025-04-05. Extremely Early Flowering and Large Grain Isogenic Japonica Rice Koshihikari Integrated with Gene e1 and GW2. https://doi.org/10.1101/2025.03.31.646432

Cite the original work for its findings. Save a collection to share your selection of sources.

KEEP EXPLORING

Related preprints

Structural variation in repeat elements is widespread in normal human tissues and in tumorigenesis

Somatic mosaicism contributes to genomic variation, yet postzygotic structural variants remain under-characterized. We performed long- and short-read WGS from multiple individuals (n=47 normal tissues; n=168 samples) and identified mosaic structural variants in all individuals and germ layers, impacting a median 285.2 kb/genome. Nearly half of breakpoints were independently validated, with tissue distributions reflecting both early and late developmental origins. Most mosaic variants were repeat-mediated and 8.3% overlapped functional elements, an enrichment compared to germline variants. To extend these analyses in samples where long-read sequencing is infeasible, we measured repeat alterations from short-read sequencing, recapitulating mosaic tissue-specific differences. We characterized tumor- and tissue- specific variation in repeats across 15 cancer types and found tumor-related repeat variation to be similar in scale to that of normal mosaic variation. Tracking repeat changes in cell-free DNA provided a noninvasive approach for tumor monitoring. Our analyses revealed widespread repeat-driven structural variation in health and disease.

genomics↗

RNA isoform-resolved multiplexed sequencing with bioorthogonal barcoding

RNA isoform dysregulation drives disease pathogenesis and is the target of FDA-approved splice-switching therapeutics. However, multiplexed sequencing methods discard splice junction information because only 3' termini are barcoded and counted. Here, we repurpose acylation and click chemistries to conjugate bioorthogonal barcodes (bobcodes) directly onto multiple internal positions along cellular RNAs. Bobcoded RNAs from multiple samples are pooled for multiplexed cDNA synthesis, during which reverse transcriptase switches from each RNA template onto its tethered bobcode with greater than 99% accuracy in species mixing experiments. Bobcode attachment intervals set cDNA insert sizes without a library fragmentation step, and priming with poly(dT) or random hexamers selects between 3'-end counting and full-length isoform capture. A bioorthogonal barcode-sequencing (BOB-seq v0.1) drug screen identifies transcriptome-wide on- and off-target RNA splicing effects and outperforms existing multiplexing RNA sequencing methods in workflow simplicity, sample-to-sample variability, and barcoding accuracy. Bobcodes add isoform resolution to scalable multiplexed RNA sequencing.

genomics↗

Structural polymorphism and population-variable coding capacity of HERV-K(HML-2) in human pangenomes

Approximately 8% of the human genome is derived from ancient retroviral infections. The most recently integrated of these endogenous retroviruses is the HERV-K(HML-2) clade, whose expression has been associated with cancer, amyotrophic lateral sclerosis, and embryogenesis. Studies of HERV expression, particularly HML-2, have relied predominantly on short-read sequencing. However, the high similarity among HML-2 proviruses prevents many short reads from being assigned uniquely to individual loci. We therefore compared haplotype-resolved long-read genome assemblies from 292 donors to resolve variation in proviral structure and coding capacity. Several loci previously thought to be fixed were structurally polymorphic. Tandem arrays occurred at 13 loci and contained up to six proviral copies in a single array. At 8q11.23, we identified a previously undescribed full-length provirus in one haplotype. All 583 other haplotypes carried a solo-LTR. We found that standard reference genomes failed to represent the coding capacity retained in many individuals, whose proviruses contained intact open reading frames despite disruptive mutations in the reference sequences. Short-read genotypes left 32.5% of the tested donor-variant pairs unresolved at sites associated with viral reading frames. These findings show why HML-2 expression must be interpreted in the context of the structural and coding alleles each individual carries.

genomics↗