Department of Plant Production, Misión Biológica de Galicia, Sede Santiago de Compostela, CSIC, Santiago de Compostela, Spain.
Maceda Nursery, Grupo Tragsa, Maceda, Ourense, Spain.
Methods Mol Biol. 2024;2759:167-181. doi: 10.1007/978-1-0716-3654-1_16.
Shoots from European chestnut and hybrids of European and Asian chestnuts can be efficiently proliferated in liquid medium by temporary immersion (TIS) using RITA and Plantform™ bioreactors. The main challenges of applying TIS to these species were the lack of growth and hyperhydricity; problem solutions included the manipulation of the hormone concentration, the explant type, immersion frequency, and a support for maintaining shoots in a vertical position. After protocol optimization, explants cultured by TIS produced more rootable shoots than explants growing in semisolid medium, enabling increased number of rooted and acclimatable shoots. In this chapter, we will describe the protocols for proliferating chestnut by TIS in RITA and Plantform™ bioreactors, together with tips for avoiding the main pitfalls of the technique. The strategies applied to chestnut can be useful for culturing other woody plants in bioreactors.
利用 RITA 和 Plantform™生物反应器中的暂态浸没（TIS）方法，欧洲栗和欧洲栗与亚洲栗的杂交品种的芽可以在液体培养基中高效增殖。将 TIS 应用于这些物种的主要挑战是生长不良和过度水合；问题的解决方案包括激素浓度、外植体类型、浸没频率的操纵，以及为保持芽处于垂直位置的支撑。在优化方案后，通过 TIS 培养的外植体比在半固态培养基中生长的外植体产生更多可生根的芽，从而增加了生根和适应环境的芽的数量。在本章中，我们将描述在 RITA 和 Plantform™生物反应器中通过 TIS 增殖栗的方案，以及避免该技术主要缺陷的技巧。应用于栗树的策略对于在生物反应器中培养其他木本植物也很有用。
