Variation in Frost Tolerance of the 1,8-Cineole-Rich Variants of the Peppermint Eucalypts, Eucalyptus Radiata and E
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J.C. Doran, A.K. Kar, J.S. Larmour and N. Reid 137 Variation in frost tolerance of the 1,8-cineole-rich variants of the peppermint eucalypts, Eucalyptus radiata and E. dives J.C. Doran1,2, A.K. Kar3, J.S. Larmour1 and N. Reid3 1CSIRO Forestry and Forest Products, PO Box E4008, Kingston, ACT 2604, Australia 2Email: [email protected] 3Ecosystem Management, School of Environmental Science and Natural Resources Management, University of New England, Armidale, NSW 2351, Australia Revised manuscript received 29 March 2005 Summary by possessing entire juvenile and mature leaves with numerous oil glands containing strongly scented essential oils. Frost tolerance is a necessary characteristic for the successful establishment of Eucalyptus radiata subsp. radiata Sieber ex DC Three subspecies are recognised in E. radiata: (1,8-cineole variant) for production of essential oil on frost-prone (i) subsp. radiata ascribing populations on the tablelands, ranges sites. The frost tolerance of seedlings from six commercial oil- and foothills of central and southern NSW, north-eastern and producing provenances of this taxon was examined using the southern central Victoria and in the Lemonthyme area in electrical conductivity method. One provenance of E. radiata northern Tasmania; subsp. robertsonii and two provenances of E. dives Schauer were (ii) subsp. sejuncta L.A.S.Johnson & K.Hill for populations on also included for comparison. To verify the validity of this method, the northern tablelands of NSW with a small extension into frost tolerance was also assessed in two field trials near Armidale southern Queensland; and on the New England Tableland of New South Wales where plants (iii) subsp. robertsonii (Blakely) L.A.S.Johnson & Blaxell for at one year from planting were scored for physical damage populations south from Orange along the western side of the attributed to the effects of frost, viz. leaf fall and stem splitting. Southern Tablelands of NSW and at higher elevations near There were consistent and significant differences in modified Canberra (Brooker and Kleinig 1999; Brooker et al. 2002). relative conductivity (RC) between provenances, and between Intra-specific variation in foliar oil composition abounds in open-pollinated families within provenances, across temperatures. E. radiata and E. dives, where up to six and five chemical variants Eucalyptus radiata subsp. radiata from Isabella River, followed respectively have been described (Johnstone 1984). The 1,8- closely by provenances from Reedy Creek and Yowrie, were the cineole-rich variants of these species are of commercial interest. most cold-tolerant, while the two E. dives provenances and E. Their oils are classed as medicinal Eucalyptus oils and are used radiata subsp. radiata from Alexandra were the least frost tolerant, for many purposes, either neat or in formulations, including based on RC values. Rankings of E. radiata subsp. radiata inhalants, liniments, ointments, aromatherapy, soaps and as provenances based on scores for stem splitting and leaf fall after disinfectants and deodorizers (Boland et al. 1991; Coppen 1995; their first winter in the field corresponded well to the RC rankings, Davis 2002). Natural populations of E. radiata and E. dives that with Reedy Creek the best performing provenance overall. Apart produce a 1,8-cineole-rich oil are limited in number and extent. from limiting frost damage through selection of hardy Commercial production centres based on natural populations provenances, the significant within-provenance family variation include the New South Wales south coast hinterland north of Bega, in frost tolerance, and the substantial variation between individuals the NSW Southern Tablelands around Tumbarumba and within families noted in these trials, suggest good potential for Rosewood, and, in the past, in the ranges outside Canberra. selection and breeding amongst hardy individuals of the best- Production in Australia of oil from 1,8-cineole-rich variants of ranked families to improve frost tolerance. E. radiata and E. dives is now relatively small: probably less than Keywords: genetic variation; frost resistance; provenance trials; essential 10 t annually. oil plants; Eucalyptus radiata; Eucalyptus dives; New South Wales Recognition that these species produce highly marketable medicinal oil has led to interest in establishing plantations of these Introduction species for oil production in Australia. More than 200 ha of E. radiata subsp. radiata oil plantation have been established near In the most recent classification of the genus Eucalyptus L’Her. Bendemeer on the New England Tableland of NSW since 1998 (Myrtaceae), Brooker (2000) placed E. radiata Sieber ex DC (M. Weber1, pers. comm. 2002). Other plantings have taken place, (narrow-leaved peppermint) and E. dives Schauer (broad-leaved peppermint) in the subgenus Eucalyptus, section Aromatica, series Radiatae. Among other attributes, these species are characterised 1Banalasta Oil Plantation Ltd, Bendemeer, NSW 2355 Australian Forestry 2005 Vol. 68 No. 2 pp. 137–143 138 Frost tolerance of peppermint eucalypts The second method was a subjective assessment of leaf fall and stem splitting among 12-mo-old seedlings in two field trials on the New England Tableland of northern New South Wales. The main trial site was at ‘Banalasta’ (30°462 S, 151°142 E, 900 m a.s.l.) near Bendemeer, with a secondary planting at Newholme (30°312 S, 151°402 E, 1000 m a.s.l.) near Armidale (Fig. 1). Species and provenances included Six provenances of E. radiata subsp. radiata, one provenance of E. radiata subsp. robertsonii and two provenances of E. dives were studied in the electrical conductivity experiment and the same six provenances of E. radiata subsp. radiata in the field trials (Table 1, Fig. 1). The material included in these studies originated from seed collected from natural populations. Most provenances were known to produce 1,8-cineole-rich essential oils. The one exception was the population of E. dives from Tumbarumba which is a piperitone- rich form (‘type’ oil). It was included in the electrical conductivity experiment for comparison. The field trials also included four seedlots of E. radiata subsp. radiata from a planted stand on the New England Tableland of unknown provenance (H. Harris2, pers. comm. 2001). Figure 1. Map of provenance location and trial sites Raising, hardening and planting of seedlings Canberra conductivity experiment usually in a farm forestry setting, on several properties along the Species, provenances and families included in this experiment Great Dividing Range in NSW and Victoria in the past 10 y. are described in Table 1. Eucalyptus radiata families were sown into plastic punnets containing a moist sowing mix of 1:1:1 of Young E. radiata subsp. radiata trees at Bendemeer suffer frost peat, perlite and washed sand during November 2000. These were damage each winter in the form of stem splitting, leaf burn and sealed in plastic bags and placed under cold-moist stratification even death. in a refrigerator at 3–5°C for two weeks. They were then The purpose of this study was to screen most of the known 1,8- transferred to a temperature-controlled glasshouse (25/18°C) for cineole variant populations of the peppermint species for frost germination under mist sprays. tolerance as young seedlings to aid in provenance selection for In the first week of January 2001, seedlings had reached the 1–2 oil plantations on frosty sites and to gain an understanding of leaf-pair stage. They were then dibbled into forestry tubes (5 cm variation in seedling frost tolerance in this group of species. A × 5 cm × 12 cm) containing a sterilised potting mix of pine bark secondary aim was to compare two methods of determining frost compost, sand, vermiculite and peat in the ratio of 5:2:2:1, and a tolerance, namely a laboratory-based method (conductivity of leaf mix of fertilisers consisting of Osmocote® 18-2.6-10 (1.6 g L–1), leachates) and subjective scoring of actual frost damage in the Osmocote® 19-2.6-10 (0.6 g L–1), dolomite (1.1 g L–1), Micromaz® field. nutrients (1 g L–1) and lime (0.3 g L–1) as well as Saturade® (1.5 g L–1). The families of E. dives were raised in like fashion Materials and methods except that washed sand was used as the sowing mix and the seed in the punnets was cold-moist stratified for four weeks. Two methods were used to estimate frost damage. The first employed the electrical conductivity method on 6-mo-old The seedlings were raised in a glasshouse for 3 mo. In April 2001, seedlings growing in the open in a nursery. This method was 10 of the most vigorous seedlings from each family were developed by Raymond et al. (1986, 1992a) and Owen and transferred to a raised bench outside. They were arranged in a Raymond (1987). It has been shown to yield the same rankings of latinised row-column design generated by the computer software seedling frost tolerance as those obtained from whole-plant studies package Alpha + (Williams and Talbot 1993) with 8 rows and 10 for E. delegatensis and E. regnans, and it was expected that there columns per replicate. Pots were spaced 6 cm apart from one would be a similar trend for the peppermint eucalypts which are another. Survival was excellent; seedling mortality was less than in the same subgenus, Monocalyptus. 2% at the time of sampling in July 2001. 2EcoResource Development Pty Ltd, Armidale, NSW 2350 Australian Forestry 2005 Vol. 68 No. 2 pp. 137–143 J.C. Doran, A.K. Kar, J.S. Larmour and N. Reid 139 Table 1. Origin of provenances (CSIRO seedlot no.) of the peppermint eucalypts in the trials at CSIRO Canberra (C) and in the field trials at Banalasta (B) and Newholme (N) on the New England Tableland of New South Wales.