In Vitro Propagation of A Cuban Arbuscular Mycorrhizal Fungal Strain

Kalyanne Fernández Suarez
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Abstract

In vitro root cultivation techniques based on modified root systems are often used in studies on Arbuscular Mycorrhizal Fungi (AMF). It is a simplified but powerful tool to investigate AMF root colonization and development of the extraradical mycelium. The aim of this study was to establish and characterize the in vitro culture of a Cuban strain of Rhizophagus irregularis (INCAM 11) by using transformed chicory roots. For that, superficially disinfected propagules of R. irregularis were co-culture with the hairy transformed chicory roots on Modified Strullu and Romand (MSR) medium during five months. Spore germination was observed 3-5 days after surface disinfection. The first contact between AMF hyphae and roots occurred 1-3 days after germination and a significant production of extensive extraradical mycelium was observed. New spore formation started within 21-25 days. After 5 months, 2000 spores could be observed per plate which were able to germinate, colonize, establish and reproduce again spores when associated to young transformed roots of chicory. The most frequent associated microorganism to the in vitro culture of INCAM 11 was isolated and identified as Paenibacillus sp.
一株古巴丛枝菌根真菌的离体繁殖
基于改良根系的离体根培养技术常用于丛枝菌根真菌(AMF)的研究。它是研究AMF根定植和根外菌丝体发育的一种简单而有力的工具。本研究的目的是利用转化的菊苣根建立古巴不规则食根菌(INCAM 11)的离体培养。为此,在改良Strullu and Romand (MSR)培养基上,将经表面消毒的不规则小檗繁殖体与毛状转化的菊苣根共培养5个月。表面消毒后3 ~ 5 d观察孢子萌发。AMF菌丝与根的第一次接触发生在萌发后1-3天,并观察到大量根外菌丝的产生。新孢子在21-25天内开始形成。5个月后,每个培养皿可观察到2000个孢子,这些孢子能够在菊苣幼芽转化根上萌发、定植、建立并再次繁殖孢子。体外培养中最常见的相关微生物为Paenibacillus sp.。
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