Factors Affecting the Regeneration, via Organogenesis, and the Selection of Transgenic Calli in the Peach Rootstock Hansen 536 (Prunus persica × Prunus amygdalus) to Express an RNAi Construct against PPV Virus
| Autor: | Cecilia Limera, Bruno Mezzetti, Silvia Sabbadini, Dania Baldoni, A. Ricci, Luca Capriotti |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Prunus spp
0106 biological sciences 0301 basic medicine Agrobacterium organogenesis Organogenesis PPV Plant Science Biology GFP 01 natural sciences Article 03 medical and health sciences Prunus Basal shoot food Prunus amygdalus Ecology Evolution Behavior and Systematics Ecology fungi Botany biology.organism_classification food.food Horticulture Transformation (genetics) 030104 developmental biology Callus RNAi QK1-989 genetic transformation Rootstock 010606 plant biology & botany |
| Zdroj: | Plants, Vol 8, Iss 6, p 178 (2019) Plants Volume 8 Issue 6 |
| ISSN: | 2223-7747 |
| Popis: | Prunus spp. is one of the most recalcitrant fruit tree species in terms of in vitro regeneration and transformation, mostly when mature tissues are used as explants. The present study describes the in vitro regeneration via indirect organogenesis, and Agrobacterium tumefaciens-mediated transformation of the peach rootstock Hansen 536 (Prunus persica × Prunus amygdalus) through the use of meristematic bulks (MBs) as starting explants. Efficient adventitious shoot regeneration was obtained when Hansen 536 MBs were cultured on an optimized medium consisting of modified McCown Woody Plant medium (WPM) enriched with 4.4 M 6-Benzyladenine (BA), 0.1 M 1-Naphthaleneacetic acid (NAA) and 6.0 g L&minus 1 plant agar S1000 (B& V). MB slices were used later as starting explants for Agrobacterium-mediated transformation to introduce an RNAi construct &ldquo ihp35S-PPV194&rdquo against PPV virus. Transgenic events were identified by both green fluorescent protein (GFP) screening and kanamycin selection at different concentrations (0, 17 or 42 M). GFP-fluorescent proliferating callus lines were selected and confirmed to stably express the ihp35S-PPV194::eGFP gene construct by molecular analysis. Although shoot regeneration from these transgenic calli has not been obtained yet, this represents one of the few examples of successful attempts in peach genetic transformation from somatic tissues, and also serves as a useful in vitro system for future gene functional analysis in peach. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|