12 research outputs found
In vitro co-cultures of Pinus pinaster with Bursaphelenchus xylophilus: a biotechnological approach to study pine wilt disease
Abstract
Main conclusion Co-cultures of Pinus pinaster with
Bursaphelenchus xylophilus were established as a
biotechnological tool to evaluate the effect of nematotoxics
addition in a host/parasite culture system.
The pinewood nematode (PWN), Bursaphelenchus xylophilus,
the causal agent of pine wilt disease (PWD), was
detected for the first time in Europe in 1999 spreading
throughout the pine forests in Portugal and recently in
Spain. Plant in vitro cultures may be a useful experimental
system to investigate the plant/nematode relationships in
loco, thus avoiding the difficulties of field assays. In this
study, Pinus pinaster in vitro cultures were established and
compared to in vivo 1 year-old plantlets by analyzing shoot
structure and volatiles production. In vitro co-cultures were
established with the PWN and the effect of the phytoparasite
on in vitro shoot structure, water content and
volatiles production was evaluated. In vitro shoots showed
similar structure and volatiles production to in vivo maritime
pine plantlets. The first macroscopic symptoms of
PWD were observed about 4 weeks after in vitro co-culture
establishment. Nematode population in the culture medium
increased and PWNs were detected in gaps of the callus
tissue and in cavities developed from the degradation of
cambial cells. In terms of volatiles main components,
plantlets, P. pinaster cultures, and P. pinaster with B. xylophilus
co-cultures were all b- and a-pinene rich. Cocultures
may be an easy-to-handle biotechnological approach
to study this pathology, envisioning the understanding
of and finding ways to restrain this highly
devastating nematode.
Keywords Maritime pine ! Monoxenic culture !
Pinewood nematode ! Relative water content ! Shoots
structure ! Volatiles
Abbreviations
BAP 6-Benzylaminopurine
DAI Days after inoculation
EPPO European and Mediterranean Plant Protectio