Plant Molecular Biology 54: 147–156, 2004.
© 2004 Kluwer Academic Publishers. Printed in the Netherlands.
Cloning and functional analyses of a gene from sugar beet up-regulated
upon cyst nematode infection
, Michael Kleine
, Carolien P. Ruyter-Spira
e M. Klein-Lankhorst
and Christian Jung
Plant Breeding Institute, Christian-Albrechts-University of Kiel, Olshausenstrasse 40, 24098 Kiel, Germany
author for correspondence; e-mail email@example.com);
Plant Research International, Postbus
16, 6700 AA Wageningen, Netherlands; present addresses:
New York State Agricultural Experimental Station,
Cornell University, Hedrick Hall, 630 West North Street, Geneva, NY 14456, USA;
Planton GmbH, Am Kiel-Kanal
44, 24106 Kiel, Germany
Received 8 November 2003; accepted in revised form 15 January 2004
Key words: Beta vulgaris, cDNA-AFLP, hairy roots, nematode resistance
The cDNA-AFLP technique was used to isolate sugar beet genes up-regulated upon infection with the beet cyst
nematode Heterodera schachtii. Hairy root cultures were obtained from resistant plants carrying a Beta procumbens
translocation as well as from a non-resistant control. mRNA was isolated from hairy root clones and sugar beet
plants infected or not with the beet cyst nematode and 8000 transcript-derived fragments (TDFs) were analysed.
One TDF was found to be differentially expressed in both materials and was further investigated. Real-time PCR
conﬁrmed that this TDF is speciﬁcally up-regulated in resistant sugar beet upon nematode infection and its full-
length cDNA was isolated. Sequence analysis suggests that the gene encodes a 317 amino acid polypeptide of
unknown function. No homology to any sequence present in the public databases could be detected. To further
elucidate its function in resistance to the beet cyst nematode, the cDNA was transformed into hairy roots of
susceptible sugar beet under the control of the 35S promoter and hairy root clones were inoculated with nematodes.
The number of developing females was signiﬁcantly reduced in 12 out of 15 clones resulting from independent
transgenic events suggesting that the gene can be used for inducing cyst nematode resistance in plants.
Abbreviations: BCN, beet cyst nematode; cDNA-AFLP, cDNA-ampliﬁed fragment length polymorphism; dai,
days after inoculation; J2, second-stage juvenile; Q-PCR, quantitative expression analysis by real-time PCR; TDF,
Plant pathogenic nematodes pose a threat to many
crops worldwide. Among them, sedentary plant-
parasitic nematodes from the genera Meloidogyne,
The sequences reported in this paper have been deposited in the
NCBI and dbEST databases. The accession numbers are incorpor-
ated in the text. The accession number of BvKi1 is AY232456. The
accession numbers of the ESTs are: TDF_6, CB280904; TDF_9,
CB280905; TDF_13, CB280906; TDF_14, CB280907; TDF_20,
CB280908; TDF_58, CB280910; TDF_65, CB280911; TDF_106,
CB280912; TDF_171, CB280913; TDF_173, CB280914;
TDF_233, CB280915; TDF_401, CB280916.
Globodera and Heterodera are the most important
ones (Sasser and Freckman, 1987). The beet cyst
nematode (BCN) Heterodera schachtii Schm. infects
sugar beet (Beta vulgaris L.) and many other plant
species. Second-stage infective juveniles (J2) penet-
rate the roots of host plants and subsequently establish
and maintain specialized feeding structures called syn-
cytia (Jones, 1981; Hussey, 1989; Wyss et al., 1992;
Sijmons et al., 1994; Williamson and Hussey, 1996).
Water and solutes are transferred into the syncytia
leading to the distortion of the entire root system. In-
fected plants show decreased growth, wilted leaves,