abstract
This study presents a new in vitro method to culture light-sensitive arbuscular mycorrhizal fungi with Lunularia cruciata. Mycothalli inoculated with Glomus clarum and Gigaspora margarita were successfully cultivated on 150 mL of dual-layer medium composed of macro- and micro-nutrients and without any of the growth promoters usually found in root organ cultures. The upper layer had the addition of 2 g L-1 of activated charcoal. Mature cultures of both fungi were obtained in less than 150 days and could be maintained viable for more than 500 days, with container size being the limiting factor for plant growth. Lunularia cruciata colonisation by both fungi showed the characteristic Paris-type morphology and was more abundant within the mycothalli's central midrib parenchyma cells than in other cells. For Gi. margarita the thallus colonisation was relatively small and shallow, suggesting a discrete distribution of the fungus within the plant and that penetration occurs mainly through new entry points of external hyphae, i.e., thallus-to-thallus penetration via appressorium. Conversely, for Gl. clartum higher colonisation was probably due to internal fungal growth along the midrib parenchyma, towards the new formed cells on the thallus' apical meristem.
keywords
DNA TRANSFORMED ROOTS; GLOMUS-PROLIFERUM; ORGAN-CULTURES; IN-VITRO; SYMBIOSIS; GLOMEROMYCOTA
subject category
Plant Sciences
authors
Fonseca, HMAC; Berbara, RLL; Pereira, ML
our authors
Maria de Lourdes Gomes Pereira
Associate Professor with Aggregationacknowledgements
The authors would like to thank Dr Ashley A.Bell for her critical and constructive comments and Alexandra Azevedo for her work in maintaining the AMF species and transgenic root collections. Finally authors acknowledged grants (1) (FCT) PEst-C/CTE/UI4035/2011 and (2) (CNPq) PEst-C/CTM/LA0011/2013.