Proceedings of the XLVI Italian Society of Agricultural Genetics - SIGA Annual Congress

Giardini Naxos, Italy - 18/21 September, 2002

ISBN 88-900622-3-1

Poster Abstract - 3.21

ISOLATION OF COLD-TOLERANT SOMACLONAL VARIANTS FROM THE SPECIES VETIVERIA ZIZANIOIDES (L.) NASH

MASSARDO D.R.*, CAPUANO M.*, VENEZIANO A.*, BORRELLI G.M.**, MAFFEI M.***, DEL GIUDICE L.*

*) Istituto di Genetica e Biofisica “Adriano Buzzati Traverso”-CNR, Via G. Marconi 10, 80125 Napoli

**) Istituto Sperimentale per la Cerealicoltura, Sezione di Foggia, S.S. 16 Km 675, 71100 Foggia

***) Dipartimento di Biologia Vegetale, Università di Torino, Viale P.A. Mattioli 25, 10125 Torino

callus, somaclonal variation, cold-tolerant plantlets, Vetiver

Vetiveria zizanioides (Maffei, 2002), a graminaceous plant native to India, is cultivated in many tropical and subtropical areas and have the capability of being both economically and ecologically important. The economic importance of the species Vetiveria zizanioides (L.) Nash depends on its ability to produce odorous roots, which can be used for the extraction of an essential oil of great economic importance. The ecological importance is due to the ability of the plant to act as a natural barrier against erosion and soil pollution. Since the Vetiver is very sensitive to temperature under 0°C, presents difficulties in cultivation in temperate and Mediterranean countries. Due to both its economical and ecological properties we were stimulated in a research program aimed to the utilization of this plant species for environmental application in our regions, through the isolation of cold-tolerant somaclonal variants, by means of in vitro selecting methods. Basal, non-differentiated, chlorophyll-free parts of leaves of Vetiveria zizanioides (L.) Nash were used as starting tissue. Cuttings were washed with sterile water, surface-sterilized in 70% ethanol for 10 sec, soaked for 15 min in a 0.1% solution of sodium hypochloride plus 2 drops of tween-20 per 500 ml and rinsed several times with sterile distilled water. Small pieces of inner leaf tissues (3-4 mm in length) were used as explants.

Callus induction was obtained from these explants cultured on Murashige and Skoog (MS) medium supplemented with 9.0 mM 2,4D, 5.7 mM IAA and 4.6 mM Kinetin (Mucciarelli et al., 1993). Calli were maintained on MS medium with the addition of 0.9 mM 2,4D, and 2.3 mM Kinetin.

In a tentative to isolate a cold-tolerant somaclonal variants, shoot formation was obtained from fast growing 14-day-old-callus on the same basal medium supplemented with 0.9 mM 2,4D and 9.3 mM Kinetin culturing in growth chambers at different night temperatures (22/24°C day, either 12°C or 15°C or 18°C night temperatures, respectively), with 14 h photoperiod 37 fluorescent TLD Philips lamps (Mucciarelli et al., 1993). Experiments on regeneration from callus cultures were carried out by placing a total of 300 ± 25 average number of callus pieces per treatment each different night temperature incubation. After 60-80 days of regenerating cultures a total of 62 plantlets deriving from embryogenic structures germinated producing green shoots, were then transferred to basal medium either with a small amount of auxin or without growth regulators, to promote root formation and the complete development of the plants.

Work is in progress to move the isolated plantlets initially in aseptic growth chamber then to experimental plots of outside environment. The plant variants presenting cold-tolerant phenotype will be selected and successively analyzed both at biochemical and molecular levels.

References

- M. Maffei (2002). Introduction to the genus Vetiveria. In “Vetiveria. The genus Vetiveria” (ed. M. Maffei), p. 1-18. Taylor & Francis, London and New York.

- M. Mucciarelli, M. Gallino, S. Scannerini, M. Maffei (1993). Callus induction and plant regeneration in Vetiveria zizanioides. Plant Cell Tiss. and Org. Cult., 35, 267-271.

This work was supported by Regione Campania, Assessorato alla Ricerca Scientifica, Legge Regionale 31.12.94, n. 41/1999, to L. Del Giudice.