The anthocyanin biosynthesis of rice is a major concern due to the potential nutritional value. Purple appears in various organs and tissues of rice such as pericarp, flower organs, leaves, leaf sheaths, internodes, ligules, apex, and stigma. At present, there are many studies on the color of rice pericarp, but the gene and mechanism of other organs such as leaves are still unclear, and the gene regulatory network of specific organ coloring has not been systematically understood. In this study, genetic analysis demonstrated that the purple leaf traits of rice were regulated by a recessive gene. The green leaf cultivar Y58S and purple leaf cultivar XianHongB were used to construct the mapping population. A set of near isogenicline (NIL) (BC3F1) was bred via crossing and back-crossing. The generations of BC3F2 appeared to separate four phenotypes, pl1, pl2, pl3, and pl4, due to the occurrence of a purple color in different organs. We constructed three bulked segregant analysis (BSA) pools (pl1–pl2, pl1–pl3, and pl1–pl4) by using the separated generations of BC3F5 and mapped the purple leaf gene plr4 to the vicinity of 27.9–31.1 Mb on chromosome 4. Subsequently, transcriptome sequencing (RNA-Seq) for pl3 and pl2 was used to analyze the differentially expressed genes in the localization interval, where 12 unigenes exhibited differential expression in which two genes (Os04g0577800, Os04g0616400) were downregulated. The two downregulated genes (Os04g0577800 and Os04g0616400) are possible candidate genes because of the recessive genetic characteristics of the purple leaf genes. These results will facilitate the cloning of plr4 and illustrate the molecular mechanisms of the anthocyanin synthesis pathway.
The ideal plant architecture is a new strategy for super high yield breeding of rice. Tiller angle is an important plant architecture character of rice. A reasonable tiller angle is a key factor for the ideal plant architecture and achieving high-yield breeding. Molecular design breeding is the most potential new direction of crop breeding in the future. The development of accurate and efficient functional molecular markers of target trait genes is crucial for molecular design breeding. The TAC1 (Tiller Angle Controlling) gene is the primary gene that regulates tiller angle in rice. This gene can be used to improve the compact plant architecture of indica and japonica rice varieties. The SNP variation from A to G at the fourth intron 3′ splicing point in TAC1 changes plant architecture. Based on the SNP variation, PM-TAC1 was successfully developed as a fluorescent functional molecular marker, via the penta-primer amplification refractory mutation system. Ninety-three rice materials were genotyped using this marker, and the marker was effectively used in rice plant architecture breeding. The successful development of this marker will contribute to the molecular breeding of rice plant architecture.
Rice (Oryza sativa) is one of the world’s major staple foods. However, stable rice production is constrained by various biotic and abiotic and stresses. Breeding and cultivation of rice varieties with resistance to multiple pathogens and environmental stresses is the most effective strategy to mitigate the adverse effect of pathogen attacks and abiotic stresses. Recently, researchers have focused on the exploitation of CRISPR/Cas9 technology to manipulate some negative defense-regulator genes to generate rice varieties with broad-spectrum resistance against rice pathogens. In this study, four negative regulator genes of rice blast, OsMads26, OsBsr-1, OsELF3-2 and OsERF922, were selected as CRISPR/Cas9 targets. By simultaneously knocking out all four genes via CRISPR/Cas9 technology, we created three mads26/bsr-1/elf3-2/erf922 quadruple knockout mutants. Our results demonstrated that all quadruple mutants exhibited much higher resistance not only to rice blast and bacterial blight but also to drought and salt stresses than the wildtype. Interestingly, grain yield of all three quadruple mutants was also drastically increased by 17.35% to 21.95%. Therefore, this study provides a novel strategy to rapidly improve rice varieties with broad-spectrum resistance to pathogens, elevated tolerance to abiotic stresses and enhanced yield potential.
The complete chloroplast genome sequence of wild cucumber (Cucumis sativus var. hardwickii) was generated by de novo assembly with low-coverage whole-genome sequence data. The 155 277 bp genome containing a pair of inverted repeats (IRs) of 25 198 bp separated by a large single-copy region of 86 618 bp and a small single-copy region of 18 263 bp. The chloroplast genome contains 130 known genes, including 89 protein-coding genes, eight rRNA genes (four kinds), and 37 tRNA genes (30 kinds). Eighteen genes are duplicated in the inverted repeat regions, 16 genes contain one intron, two genes, and one ycf contain two introns. Phylogenomic analysis showed that C. sativus var. Hardwickii is closely related to C. sativus and C. hystrix.
Rice blast and bacterial blight cause severe harm to rice production, and the breeding of resistant varieties guarantees the safety of rice production. Meanwhile, multigene pyramiding breeding based on molecular marker-assisted selection is a crucial approach for rice breeding to combat multiple diseases. This study aimed to develop accurate and efficient PARMS markers for rice blast resistance genes Pita, Pigm, and Pi2, and bacterial blight resistance gene Xa23. A systematic genotyping analysis of the resistant alleles of these 4 genes was performed on 384 major cultivated varieties in production. The results showed that only 5.21% of the varieties harbored more than two resistant alleles simultaneously. Using traditional breeding strategies in combination with the developed PARMS markers, the high-quality three-line male sterile line Ruanfeng A (pyramiding Pita and Pigm) and the strong restorer line Gui 610 (pyramiding Pi2 and Xa23) were bred. Crossing these lines produced a new hybrid rice variety, Ruanfengyou 610. Ruanfengyou 610 pyramids 4 resistance genes (Pita/Pigm/Pi2/Xa23), exhibits resistance to both rice blast and bacterial blight, has prominent heterosis and excellent grain quality, and has strong application potential, which is of great significance for ensuring the safety of rice production.