Plant Cell, Tissue and Organ Culture (PCTOC)

, Volume 118, Issue 3, pp 519–530 | Cite as

In vitro co-culture of Solanum tuberosum hairy roots with Meloidogyne chitwoodi: structure, growth and production of volatiles

  • Jorge M. S. Faria
  • Inês Sena
  • Carla M. Maleita
  • Inês Vieira da Silva
  • Lia Ascensão
  • Isabel Abrantes
  • Richard N. Bennett
  • Manuel Mota
  • A. Cristina Figueiredo
Original Paper

Abstract

Meloidogyne spp., commonly known as root-knot nematodes (RKNs), are economically important plant sedentary endoparasites that cause galls on susceptible hosts. The Columbia root-knot nematode (CRKN), M. chitwoodi, is a quarantine A2 type pest by the European and Mediterranean Plant Protection Organization since 1998. This nematode has been found associated with economically important crops such as potato and tomato, causing severe damage and making the agricultural products unacceptable for the fresh market and food processing. In vitro co-culture of host and parasite offers an advantageous experimental system for studying plant-RKN interactions. The structure, growth and production of volatiles of Solanum tuberosum hairy roots (HR) and of S. tuberosum HR/CRKN co-cultures were compared. HR were induced by inoculation of aseptic potato tuber segments with Rhizobium rhizogenes. Co-cultures were initiated by inoculating HR with sterilized CRKN eggs. Infection with CRKN induced the RKN symptomatology in the HR and several nematode life stages were observed by light and scanning electron microscopy. Potato HR and HR/CRKN co-cultures exhibited similar growth patterns, evaluated by measuring fresh and dry weight and by the dissimilation method. Volatiles, isolated by distillation–extraction and analyzed by gas chromatography (GC) and gas chromatography coupled to mass spectrometry, revealed that palmitic acid (37–52 %), n–pentadecanal (10–16 %) and linoleic acid (2–16 %) were the main constitutive components of S. tuberosum HR, and of the HR/CRKN co-cultures (24–44, 8–22 and 4–18 %, respectively). S. tuberosum HR/CRKN co-cultures can be considered a suitable biotechnological tool to study RKN infection mechanism by mimicking what occurs under field conditions.

Keywords

Plant biotechnology Columbia root-knot nematode Potato Hairy root structure Root gall structure Volatiles 

Abbreviations

µ

Specific growth rate

CRKN

Columbia root-knot nematode (Meloidogyne chitwoodi)

DAI

Days after inoculation

dt

Doubling time

DW

Dry weight

EPPO

European and Mediterranean Plant Protection Organization

FW

Fresh weight

GC

Gas chromatography

GC–MS

Gas chromatography coupled to Mass Spectrometry

HgCl2

Mercuric chloride

HR

Hairy roots

J2

Nematode second-stage juveniles

LM

Light microscopy

NaOCl

Sodium hypochlorite

OsO4

Osmium tetroxide

PAS

Periodic acid–Schiff’s reagent

RI

Retention index

RKN

Root-knot nematode

SEM

Scanning electron microscopy

SH

Schenk and Hildebrandt medium

t

Trace

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Jorge M. S. Faria
    • 1
  • Inês Sena
    • 1
  • Carla M. Maleita
    • 2
  • Inês Vieira da Silva
    • 1
  • Lia Ascensão
    • 1
  • Isabel Abrantes
    • 3
  • Richard N. Bennett
    • 4
  • Manuel Mota
    • 5
    • 6
  • A. Cristina Figueiredo
    • 1
  1. 1.Universidade de Lisboa, Faculdade de Ciências de Lisboa, DBV, IBB, Centro de Biotecnologia Vegetal, C2, Campo Grande, 1749-016 Lisboa, PortugalLisbonPortugal
  2. 2.CIEPQPF, Departamento de Engenharia Química, Universidade de Coimbra, 3030-790 Coimbra, PortugalCoimbraPortugal
  3. 3.IMAR-CMA, Departamento de Ciências da Vida, Universidade de Coimbra, 3004-517 Coimbra, PortugalCoimbraPortugal
  4. 4.Universidade de Trás-os-Montes e Alto DouroVila RealPortugal
  5. 5.NemaLab, ICAAM-Instituto de Ciências Agrárias e Ambientais MediterrânicasUniversidade de ÉvoraÉvoraPortugal
  6. 6.INIAV/Unidade Estratégica de Investigação e Serviços de Sistemas Agrários e Florestais e Sanidade VegetalOeirasPortugal

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