Anther culture (AC) is a valuable technique in rice breeding. However, the genetic mechanisms underlying anther culturability remain elusive, which has hindered its widespread adoption in rice breeding programs. During AC, microspores carrying favorable alleles for AC are selectively regenerated, leading to segregation distortion (SD) of chromosomal regions linked to these alleles in the doubled haploid (DH) population. Using the AC method, a DH population was generated from the japonica hybrid rice Shenyou 26. A genetic map consisting of 470 SNPs was constructed using this DH population, and SD analysis was performed at both the single- and two-locus levels to dissect the genetic basis underlying anther culturability. Five segregation distortion loci (SDLs) potentially linked to anther culturability were identified. Among these, SDL5 exhibited an overrepresentation of alleles from the female parent, while SDL1.1, SDL1.2, SDL2, and SDL7 displayed an overrepresentation of alleles from the male parent. Furthermore, six pairs of epistatic interactions (EPIs) that influenced two-locus SDs in the DH population were discovered. A cluster of genetic loci, associated with EPI-1, EPI-3, EPI-4, and EPI-5, overlapped with SDL1.1, indicating that the SDL1.1 locus may play a role in regulating anther culturability via both additive and epistatic mechanisms. These findings provide valuable insights into the genetic control of anther culturability in rice and lay the foundation for future research focused on identifying the causal genes associated with anther culturability.

The development of a new salt–alkaline-tolerant hybrid japonica rice is crucial for enhancing japonica rice supply and ensuring national food security. Utilizing molecular marker-assisted selection (MAS) technology combining Kompetitive Allele-Specific PCR (KASP) markers and a gene breeding chip, the salt-tolerant gene SKC1 was introgressed into a rice genotype Fan 14. This led to the development of Shenyanhui 1, a new high-quality, strongly heterotic, and salt-tolerant japonica restorer line. Subsequently, the high-quality, salt-tolerant japonica three-line hybrid rice variety Shenyanyou 1 was developed by crossing the BT-type japonica cytoplasmic male sterile (CMS) line Shen 21A with the restorer line Shenyanhui 1. Shenyanyou 1 carries the major salt tolerance gene SKC1, exhibiting excellent salt tolerance with seedling stage salt tolerance reaching level 5. Under precise salt tolerance evaluation throughout its growth cycle, Shenyanyou 1 achieved a yield of 3640.5 kg/hm², representing an extremely significant increase of 20.7% over the control variety Yandao 21. Shenyanyou 1 exhibits superior grain quality, meeting the Grade 3 high-quality rice standards issued by the Ministry of Agriculture. Shenyanyou 1 has good comprehensive resistance, aggregating rice blast resistance genes such as Pi2, Pita, Pizt and LHCB5, bacterial blight resistance genes Xa26/Xa3, stripe blast resistance gene STV11, semi-dwarf gene Sdt97, nitrogen-efficient utilization gene NRT1.1B, the light repair activity enhancement gene qUVR-10, the cold resistance gene qLTG3-1, and the iron tolerance gene OsFRO1. It has good resistance to biotic and abiotic stresses. This paper details the breeding process, key agronomic traits, salt tolerance, yield performance, and grain quality characteristics of Shenyanyou 1.

Fusarium fujikuroi is the primary causal agent of rice bakanae disease, which can lead to substantial yield losses. Developing a rapid, highly specific, and accurate method for detecting F. fujikuroi is crucial for effective surveillance, prevention, and control of rice bakanae disease. In this study, a novel detection assay, RPA-Cas12a-F, was developed by integrating recombinase polymerase amplification (RPA) and Cas12a for the detection of F. fujikuroi. This assay demonstrated a limit of detection (LOD) of 1 copy/μL of reference plasmid or 0.1 fg/μL of F. fujikuroi genomic DNA (gDNA). Furthermore, to enable on-site detection, the RPA-Cas12a technique was combined with a lateral flow strip (LFS) for visual readout, thereby developing the RPA-Cas12a-LFS assay. The LOD of the RPA-Cas12a-LFS assay was 1000 copies/μL of plasmid or 10 fg/μL of F. fujikuroi gDNA. The RPA-Cas12a-based assays developed in this study enable rapid, highly accurate, sensitive, and specific detection of F. fujikuroi, making them a promising tool for on-site detection without the need for expensive equipment and time-consuming methodologies.

The acetohydroxyacid synthase (AHAS) is an essential enzyme involved in branched amino acids. Several herbicides wither weeds via inhibiting AHAS activity, and the AHAS mutants show tolerance to these herbicides. However, most AHAS mutations are residue substitutions but not residue deletion. Here, residue deletion was used to engineering the AHAS gene and herbicide-tolerant rice. Molecular docking analysis predicted that the W548 of the AHAS was a residue deletion to generate herbicide tolerance. The AHAS-ΔW548 protein was generated in vitro to remove the W548 residue. Interestingly, the deletion led to the tetramer dissociation of the AHAS, while this dissociation did not reduce the activity of the AHAS. Moreover, the W548 deletion contributed to multi-family herbicides tolerance. Specially, it conferred more tolerance to sulfometuron-methyl and bispyribac-sodium than the W548L substitution. Further analysis revealed that AHAS-ΔW548 had the best performance on the sulfometuron-methyl tolerance compared to the wild-type control. Over-expression of the AHAS-ΔW548 gene into rice led to the tolerance of multiple herbicides in the transgenic line. The T-DNA insertion and the herbicide treatment did not affect the agronomic traits and yields, while more branched-chain amino acids were detected in transgenic rice seeds. Residue deletion of W548 in the AHAS could be a useful strategy for engineering herbicide tolerant rice. The increase of branched-chain amino acids might improve the umami tastes of the rice.

Rapid Detection of Rice Fragrance Allele badh2-E7 by Recombinant Polymerase Amplification (RPA)

The advancement of hybrid japonica rice is pivotal for securing japonica rice supplies and bolstering food security. To address prevalent issues such as inconsistent yields, subpar rice quality, and inadequate seed production in existing cultivars, Shenyou R3 was developed using advanced high-density rice gene chip technology, which is characterized by the expression of specific genes. This late-season, premium aromatic variety, characterized by a popcorn-like aroma, was bred by the Crop Breeding and Cultivation Research Institute of the Shanghai Academy of Agricultural Sciences. Shenyou R3 incorporates superior genes such as badh2-E7, Pi2, Xa21, Sdt97, and Hd17, among which, badh2-E7 and Hd17 are inherited from the maternal line, while Pi2, Xa21, and Sdt97 are inherited from both the maternal and paternal lines. Shenyou R3 offers high-quality rice that adheres to national premium grade 2 standards, with level 1 resistance to blast disease, and yields surpassing the control variety Huayou 14 by over 5% in 2022 Shanghai trials. The new hybrid japonica rice Shenyou R3 has high yield potential and nitrogen utilization efficiency. This paper elaborates on the molecular marker-assisted selection process, key traits, quality metrics, and yield performance of Shenyou R3, while also outlining essential cultivation practices.