Cloning and functional analyses of a gene from sugar beet up-regulated upon cyst nematode infection.
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.
Samuelian
Suren
S
Plant Breeding Institute, Christian-Albrechts-University of Kiel, Olshausenstrasse 40, 24098 Kiel, Germany.
Kleine
Michael
M
Ruyter-Spira
Carolien P
CP
Klein-Lankhorst
René M
RM
Jung
Christian
C
D000818:Animals; D027461:Beta vulgaris / Q000235:genetics* / Q000469:parasitology; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000737:chemistry / Q000235:genetics; D005260:Female; D020869:Gene Expression Profiling; D018506:Gene Expression Regulation, Plant; D005816:Genetic Complementation Test; D007113:Immunity, Innate / Q000235:genetics; D008969:Molecular Sequence Data; D021141:Nucleic Acid Amplification Techniques; D010935:Plant Diseases / Q000235:genetics* / Q000469:parasitology; D010940:Plant Proteins / Q000235:genetics*; D018517:Plant Roots / Q000235:genetics / Q000469:parasitology; D030821:Plants, Genetically Modified; D017422:Sequence Analysis, DNA; D013997:Time Factors; D014158:Transcription, Genetic / Q000235:genetics; D014431:Tylenchoidea / Q000254:growth & development*; D015854:Up-Regulation
D018076:DNA, Complementary; D010940:Plant Proteins