Sugarcane white leaf (SCWL)-diseased sugarcane plants collected from Udornthani Province, in north-eastern Thailand, were the source for tissue culture experiments. Explants from axillary buds, meristem tips, and leaves grew optimally in Murashige-Skoog medium containing 0.5 mg/l α-naphthaleneacetic acid, 0.5 mg/l 6-benzylaminopurine, and 15% coconut water. Callus development and shoot/root proliferation were more rapid in cultures from diseased than from healthy plants. Disease symptoms continued for 6 years after culture initiation, and SCWL phytoplasma persisted, as confirmed by polymerase chain reaction using both 16S rDNA and 16S-23S rDNA primers. Phytoplasmas in the cultured plantlets were transmissible by grafting to sugarcane and periwinkle, and by feeding of the leafhopper vector Matsumuratettix hiroglyphicus to sugarcane. Although 50% of the plantlets were killed by oxytetracycline at 500 mg/ml, 70–100% of plantlets grown with 200–500 mg/ml oxytetracycline showed symptom remission through 5–8 subcultures. Typical phytoplasma-like bodies, visible by electron microscopy in sieve tubes of untreated diseased plantlets, were absent in antibiotic-treated plantlets. Thus, tissue culture provides a convenient and reliable in vivo system for investigation of SCWL phytoplasma.
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This work was supported by a grant from the Thailand Research Fund and Khon Kaen University, and by the Oklahoma Agricultural Experiment Station project # 2052.
Communicated by H. Judelson
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Wongkaew, P., Fletcher, J. Sugarcane white leaf phytoplasma in tissue culture: long-term maintenance, transmission, and oxytetracycline remission. Plant Cell Rep 23, 426–434 (2004). https://doi.org/10.1007/s00299-004-0847-2
- Sugarcane tissue culture
- SCWL phytoplasma
- Graft transmission
- Insect transmission
- Oxytetracycline treatment