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Isolated microspore culture has been implemented in breeding programs to produce doubled haploid (DH) lines and thus accelerates the breeding process. However, low microspore embryogenesis frequency in flowering Chinese cabbage remains a key obstacle to the practical application of this technique. This study aimed to establish an efficient microspore culture protocol for flowering Chinese cabbage that would be applied for heterosis breeding. Microspores of five genotypes, 19AY05, 19AY06, 19AY10, 19AY12, and 19AY15, were successfully induced to produce embryos in NLN-13 medium. Microspores of two genotypes, 19AY05 and 19AY15, were cultivated in NLN-13 medium supplemented with different concentrations (0, 0.01, 0.05, 0.1, or 0.2 mg·L−1) of compound sodium nitrophenol (sodium nitrophenol, 5-nitrophenol) to enhance microspore embryogenesis and plant regeneration without an intervening callus phase. The results showed that 0.05 ~ 0.1 mg· L−1 sodium nitrophenol and 0.01 ~ 0.2 mg· L−1 of 5-nitrophenol significantly promoted the induction of microspore embryogenesis of two genotypes, and the best concentrations required for different genotypes are different. Moreover, 0.1 mg· L−1 sodium nitrophenol can significantly increase the plant regeneration rate of the two genetypes. The 5-nitrophenol at 0.01 mg·L−1 significantly increased rate of embryos directly convert to plant in 19AY15. In addition, the average doubled haploid rates in the five genotypes were close to 63%. Horticultural traits of DH lines from 19AY05 were identified and all of them were self-incompatible lines. They showed a high uniformity and consistency that can be directly used for hybrid breeding. Furthermore, the hybrid combination was prepared with the selected DH lines and the Guangdong nucleus genic sterile line GMS019 to screen the excellent hybrid combination for the flowering Chinese cabbage breeding program. This method accelerates the application of microspore culture in hybrid breeding of flowering Chinese cabbage.
相似文献Arbuscular mycorrhizal (AM) fungi can form symbiosis with 90% of the vascular plants and play important roles in ecosystem. To realize the AM fungal colonization at different succession stages in saline-alkali land and screen AM fungi species with great functions, roots and soil samples were collected from the three succession stages of Songnen saline-alkali grassland. The soil properties and AM fungal colonization were measured, and the fungus distributed extensively in three stages was annotated by sequencing for AML1/AML2 target, subsequently, maize was selected as the host to verify its colonization. The results showed that the soil properties improved with the succession of saline-alkali grassland. The plants’ communities of the three stages could be colonized by AM fungi, and the colonization rate of Leymus chinensis (the third stage) ranged from 66.67% to 100%, Puccinellia tenuiflora (the second stage) ranged from 50% to 80%, while the Suaeda glauca (the first stage) was only 35%–60%. Glomeraceae sp1 was identified as the dominant AM fungi species which occurred frequently in the succession of saline-alkali land with the isolation frequency, relative abundance, and importance value of 100%, 18.1%, and 59.1%, respectively. The colonization rate of Glomeraceae sp1 in maize ranged from 80% to 87% and similar mycorrhizal characteristics were detected in the roots of P. tenuiflora, S. glauca, and L. chinensis, indicating that Glomeraceae sp1 colonized the samples in the field. The correlation matrix indicated that colonization rate, colonization intensity, and vesicle abundance were closely related to soil conditions most, and they were related significantly to all the soil properties except cellulase activity. Besides, redundancy analysis (RDA) showed that soil properties drove the changes of AM fungal colonization and sporulation. These results will provide theoretical support for realizing the relationship between AM fungal colonization and soil conditions, and also for the exploration of AM fungi species with great functions.
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