Micropropagation, estimation of DNA methylation during multiplication cycles and mycorrhization of seed-derived Dendrocalamus asper (Schultes f.) Backer ex Heyne

• Published in: Plant cell, tissue and organ culture tissue and organ culture, 2023-10, Vol.155 (1), p.41-56
• Main Authors: Himmen, Fernanda Duarte Araújo, de Souza, Francisco Adriano, de Araújo Silva-Cardoso, Inaê Mariê, de Souza, André Lúis Xavier, Scherwinski-Pereira, Jonny Everson
• Format: Article
• Language: English
• Subjects: Bamboo; Benzyladenine; Biomedical and Life Sciences; Colonization; Culture media; Dendrocalamus asper; Deoxyribonucleic acid; DNA; DNA methylation; Hyphae; Kinetin; Life Sciences; Micropropagation; Optimization; Original Article; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plant Sciences; Plantlets; Shoots; Spores; Subculture; Sucrose
• ISSN: 0167-6857; 1573-5044
• DOI: 10.1007/s11240-023-02547-x

Considering the economic significance of Dendrocalamus asper , a woody bamboo species, the objective of this study was to establish an in vitro micropropagation and mycorrhization protocol, and to analyze the DNA methylation dynamics during the multiplication stage of plants of this species. For shoot proliferation, meta - topolin ( m T), 6-benzylaminopurine (BAP) and kinetin (Kin) were evaluated at different concentrations (0, 3.3, 6.6 and 13.3 µM) during three monthly subcultures. For in vitro mycorrhization, Rhizoglomus clarum inoculated in liquid medium was used. Murashige and Skoog (MS) and Strullu and Romand (MSR) culture media were tested with different modifications, such as reduced phosphorus, sucrose, and salts. The evidence of root colonization (spores, hyphae, or vesicles) was assessed. For micropropagation, it was verified that m T was as efficient as BAP for the in vitro multiplication of D. asper shoots. There was also a substantial reduction in the number of shoots in the third subculture, which coincided with a significant decrease in DNA methylation evaluated via ELISA, unprecedented analysis carried out during bamboo micropropagation. As for mycorrhization, colonization was observed only in plants grown on MSR (16% of the plants) and MS/2 medium with 25% of the total amount of sucrose (25% of the plants). These results will support the development of efficient in vitro mycorrhization protocols for D. asper and, consequently, the optimization of the quality of the micropropagated plantlets. Key message In this study a protocol for micropropagation and in vitro mycorrhization of Dendrocalamus asper was developed and DNA hypomethylation was found to be associated with the decrease in multiplication rate of shoots after successive subcultures.