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Leaf-induced callus formation in two cultivars: hot pepper 'CM334' and bell pepper 'Dempsey'.

ABSTRACT: Pepper (Capsicum annuum), one of the most economically important vegetables of the Solanaceae family, is cultivated worldwide. To apply versatile genome-editing tools to a pepper genome for precise molecular breeding, an in vitro regeneration protocol is indispensable and callus formation is an essential step in the regeneration of pepper. Here, we show that calli were successfully induced from young leaves (3-4 cm) of pepper plants, the hot pepper C. annum 'CM334' ('CM334') and bell pepper C. annum 'Dempsey' ('Dempsey'), grown on soil for less than 7 weeks. The excised leaf segments of 'CM334' produced white calli in B5 medium containing 3% sucrose (3S), 2 mg/L 6-benzylaminopurine (2BAP), and 1 mg/L α-naphthalene acetic acid (1NAA). The calli were able to proliferate in B5 3S 2BAP medium supplemented with 2-morpholinoethanesulphonic acid (MES) and 1.5 mg/L NAA (1.5NAA). The excised leaf segments of 'Dempsey' produced light-yellow and friable calli in MS medium supplemented with B5 vitamins (MSB5), 3S and 1 mg/L 2,4-dichlorophenoxyacetic acid (1 2,4D), and the calli were also maintained in the same medium. Our findings establish the conditions for leaf-derived callus formation, which is the basis for regeneration of whole plants for two different pepper cultivars, for obtaining stable protoplasts, and eventually for applying genome-editing tools to improve the quality of peppers.

SUBMITTER: Kim H

PROVIDER: S-EPMC6619979 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:The plant microbiome is shaped by plant development and microbial interaction. Fungal pathogens infecting bell pepper plants may fluctuate across the growing seasons. Dynamic fluctuation of the microbiome and fungal pathogens in bell pepper plants is poorly understood, and the origin of fungal pathogens causing fruit rot and leaf wilt has been barely investigated. In this study, we used amplicon sequencing (i.e., 16S rRNA and internal transcribed spacer [ITS] sequencing) to explore the compositional variations of the microbiome in bell pepper plants and studied the fluctuation of fungal pathogens across the growing seasons. Co-occurrence network analysis was applied to track the origin and dissemination route of fungal pathogens that infected bell pepper plants. ITS and 16S rRNA sequencing analyses demonstrated that fungal pathogens infecting fruits and leaves probably belonged to the Penicillium, Cladosporium, Fusarium, and unclassified_Sclerotiniaceae genera rather than one specific genus. The dominant fungal pathogens were different, along with the development of bell pepper plants. Both plant development and fungal pathogens shaped microbial communities in bell pepper plants across the growing seasons. Fungal pathogens decreased species richness and diversity of fungal communities in fungus-infected fruit and leaf tissues but not the uninfected stem tissues. Bacterial metabolic functions of xenobiotics increased in fungus-infected leaves at a mature developmental stage. Competitive interaction was present between fungal and bacterial communities in leaves. Co-occurrence network analysis revealed that the origins of fungal pathogens included the greenhouse, packing house, and storage room. Niche differentiation of microbes was discovered among these locations. IMPORTANCE Bell peppers are widely consumed worldwide. Fungal pathogen infections of bell peppers lead to enormous economic loss. To control fungal pathogens and increase economic benefit, it is essential to investigate the shifting patterns of the microbiome and fungal pathogens in bell pepper plants across the growing seasons. In this study, bell pepper plant diseases observed in fruits and leaves were caused by different fungal pathogens. Fungal pathogens originated from the greenhouse, packing house, and storage room, and niche differentiation existed among microbes. This study improves the understanding of dynamic fluctuation and source of fungal pathogens infecting bell pepper plants in the farming system. It also facilitates precise management of fungal pathogens in the greenhouse.

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Leaf-induced callus formation in two cultivars: hot pepper 'CM334' and bell pepper 'Dempsey'.

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