Essential Oils - RIRDC Completed Projects in 2005-2006 & Research in Progress as at June 2006

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RIRDC Completed Projects in 2005-2006 & Research in Progress as at June 2006
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1.4 Essential Oils - Completed Projects
PROJECT No	PROJECT TITLE	RESEARCHER	PHONE	ORGANISATION

Development and Improvement of Products

DAQ-328A	A literature review and assessment of Morinda citrifolia (Noni)	Ms. Helen MacPherson	(07) 3406 8589	Department of Primary Industries and Fisheries (Qld)
UCS-31A	Improvement of lavender varieties by manipulation of chromosome number	Dr. Nigel Urwin	(02) 6933 2450	Charles Sturt University
UT-39A	Development of an internationally competitive saffron industry	Prof. Robert Menary	(03) 6226 2723	University of Tasmania

Development and Improvement of Products

Project Title:	*A literature review and assessment of Morinda citrifolia* (Noni)**
RIRDC Project No.:	DAQ-328A
Researcher:	Ms Helen MacPherson
Organisation:	Department of Primary Industries (Qld) Centre for Food Technology 19 Hercules Street, HAMILTON QLD 4007
Phone:	(07) 3406 8589
Fax:	(07) 3406 8677
Email:	helen.macpherson@dpi.qld.gov.au
Objectives	To investigate the potential for a new value adding industry by conducting an extensive literature review on the plant Morinda citrifolia (Noni) and its juice extract. The objectives are to assess: ·1 market opportunities and examine potential returns to Australian producers ·2 the broad range of supposed therapeutic effects ·3 the fermentation process for quality control ·4 existing agronomic best practices
Background	Noni grows widely throughout the Pacific and is native to Australia. There are many claims as to the efficacy of noni juice from its use by traditional healers but there appears to be little in the way of scientifically reproducible studies. A group of growers in Far North Queensland have commenced planting noni trees and have formed an association (Australian Noni Producers Inc.) to initiate the growth of this opportunity to produce plant extracts from both the native and exotic varieties of this species. The association has identified that information is required to progress the development of this industry.
Research	By searching, compiling and reviewing the extensive existing literature on the plant Morinda citrifolia (noni) and its juice extract, the researchers were able to assess the broad range of agronomic best practices, the fermentation process, supposed therapeutic effects and the market opportunities and potential returns to Australian producers.
Outcomes	Agronomic best practices from both international and local sources have been reviewed and extensive information provided on the botany, ecology and cultural practices of the plant Morinda citrifolia (noni). It has been demonstrated that the basic mechanisms and associated microflora of the reported noni juice fermentation process is not well understood. Further research is required to identify the mechanisims and microflora involved in the reported fermentation process to develop commercial process parameters, potential processing aids (starter cultures) and industry standards. In vitro research and limited animal experiments have shown that noni has antimicrobial, anti-cancer, antioxidant, anti-inflammatory, analgesic and cardiovascular activity. However more studies are needed to identify the specific compounds and their mechanisms of action in relation to potential health benefits.
Implications	Based on the results of this review of existing literature, it is clear that further scientific investigations are required to address important issues in agronomic and processing processes, possible health benefits and marketability of the noni fruit as a potential crop in Australia.

Development and Improvement of Products

Project Title:	Improvement of lavender varieties by manipulation of chromosome number
RIRDC Project No.:	UCS-31A
Researcher:	Dr. Nigel Urwin
Organisation:	Charles Sturt University School of Agriculture Locked Bag 588 WAGGA WAGGA NSW 2678
Phone:	(02) 6933 2450
Fax:	(02) 2933 2812
Email:	nurwin@csu.edu.au
Objectives	This project aimed to breed improved varieties of Lavandula angustifolia (lavender) and Lavandula x intermedia (lavandin) both of which are grown commercially for their essential oil, dried and cut flowers. Plant tissue culture techniques and the use of mitotic spindle inhibitors will be used to induce polyploidy. The specific outcomes of the proposed research are as follows: Polyploid L. angustifolia and L. x intermedia cultivars, with improved oil yield and vigour. Double haploid L. angustifolia and fertile L. x intermedia cultivars, which can be propagated by seed.
Background	Lavender, (L. angustifolia) is grown for its essential oil, which is used in the perfume industry. Lavandin, (L. x intermedia) cultivars are sterile hybrids of the species L. angustifolia and L. latifolia. These are also grown for their oil, which is of lower quality but used in various toiletries. Lavender is also grown for cut and dried flowers and several of the 39 species within the genus are grown as ornamentals. The Australian lavender industry produces less than 1% of total oil produced worldwide but recently there has been increased interest in lavender as an alternative crop given the increased demand for essential oils particularly in complimentary therapies. Little effort has been put into breeding improved cultivars of lavender for commercial oil production. Polyploidy induction (producing plant with more than two sets of chromosomes) in agricultural and horticultural crops is commonly used for plant improvement producing bigger plants with increased vigour. L. angustifolia cultivars which produce the highest grade oils are small plants with smaller flowers in comparison to L. x intermedia cultivars. L. x intermedia cultivars produce 3-5 times more oil than L. angustifolia cultivars. Polyploid varieties of L. angustifolia and L. x intermedia are likely to be larger, have bigger flowers and consequently yield more oil. The new germplasm would be useful for future breeding. This project aims to give Australian lavender growers access to lavender cultivars improved with respect to oil yield and quality. The Australian horticulture industry will also benefit from marketing these plants internationally. For example a single ornamental lavender (1 million plants) sold in the US under licence with a 10cent royalty per plant would yield >AUD$250,000 in income to Australia alone. If these cultivars are adopted for commercial oil production worldwide the benefit from royalties alone is likely to be at least 10-fold greater. It is hoped the availability of these plants will encourage growth in the Australian lavender industry.

Research	Using standard plant propagation techniques, plant tissue culture techniques and colchicine (a mitotic spindle inhibitor) we have produced varieties of lavender and lavandin with more than the usual two sets of chromosomes (polyploids). Using a variety of methods we have begun to characterise these plants in terms of their genome size and morphological characteristics, particularly those which will affect their productive capacity such as flower size, oil gland size, oil yield and quality.
Outcomes	Fertile tetraploid L. angustifolia varieties producing perfume quality oil, infertile polyploid (probably triploid) L. angustifolia and L. x intermedia varieties with improved vigour, larger flowers, potentially higher oil yield and improved oil quality.
Implications	The Australian lavender industry will be encouraged to adopt and benefit from the new germplasm.
Publications	Urwin N, Horsnell, J and Moon T (2005) Improvement of Lavender by Manipulation of Chromosome number. Lavender Bag, 23:5-10 Urwin N (2004) Breeding New Lavenders. The Australian Lavender Growers Association (TALGA) Annual conference, Halls Gap, Victoria September 2004. Oral presentation.

Development and Improvement of Products

Project Title:	Development of an internationally competitive saffron industry
RIRDC Project No.:	UT-39A
Researcher:	Prof Robert Menary
Organisation:	University of Tasmania School of Agricultural Science Private Bag 54 HOBART TAS 7001
Phone:	(03) 6226 2723
Fax:	(03) 6226 7609
Email:	r.menary@utas.edu.au
Objectives	·1 To meet ISO standard for Tas-Saff saffron. ·2 To identify and design improved saffron drying technology (This was changed with the agreement of RIRDC following request in the 2004 Milestone Report). ·3 To identify potential fragrance products from waste flowers and alkaloids in dormant corms. ·4 Work closely with management and staff of Tas-Saff Pty.Ltd. to focus research and implement recommendations.
Background	Tas-Saff is the only significant producer of saffron in Australia and have been marketing product locally for over 10 years. The company is aiming to achieve import replacement by capturing 30% of the domestic market by 2013 as well as access export markets, but is competing with cheaper products from countries with much lower labour costs. The Industry Partners have therefore identified the need to differentiate their product as being of premium quality. To this end they have sought to both analytically demonstrate the quality of their current product relative to competitors and determine how their post harvest processing methods might be altered to further improve this quality and/or develop new products from the crop and thus provide competitive advantage.
Research	ISO standard and alternative analytical measurement techniques were applied and compared for Tas-Saff samples from different local growers and imported products. These techniques were also used to investigate the stigma drying process to experimentally determine the optimum conditions for retention of the colour and production of aroma components in the spice. Distillation and extraction techniques, combined with organoleptic and chemical analysis, were used to investigate potential fragrance products from the waste flower parts. Extraction and chemical analysis methods were also used to investigate the reported presence of commercially valuable alkaloids in the dormant Crocus corms.

Outcomes

Local saffron products generally compared well to imported products for colour, but not for aroma contents. The current drying technique (employing air-flow at moderate temperature) does not produce optimum aroma development and/or retention and carries the risk of insufficient drying leading to colour degradation in storage. It was shown that by drying at a significantly higher temperature, without air-flow, but in an elevated humidity atmosphere; highly significant increases in the content of the main aroma compound were produced. Furthermore, these conditions could enhance the retention and/or availability of the main pigments in the saffron.

A waxy solvent extract from the waste flower parts was identified as having potential as a new commercial fragrance product although a small addition of saffron to these flowers may be required to give the extract sufficient aroma impact. Distillation and corm extraction did not identify potential products.

Implications

This project has shown that by applying the appropriate drying conditions, it is possible to significantly and consistently improve the quality of the local product such that it is superior to that produced by overseas competitors. To this end it is recommended that the local producers use fan-less ovens, either modified or specifically constructed, that are sufficiently enclosed to allow enhanced humidity from a container of water placed in the chamber.

This project also found that there is potential for value adding to the saffron crop.

1.4 Essential Oils - Research in Progress
PROJECT No	PROJECT TITLE	RESEARCHER	PHONE	ORGANISATION

Development and Improvement of Products

BRA-4A	Development and economic assessment of Artemisia for production of anti-malarial extract	Dr. Rowland Laurence	(03) 6425 5888	Botanical Resources Australia Ltd
UNC-21A	Analysis of Dodonea viscosa as a potential remedy for anti-inflammatory and antibacterial applications	Dr. Douglas Stuart	(02) 4348 4124	The University of Newcastle

Improved Production Systems

DAQ-329A	Commercial production of milkweed, a plant with anticancer properties	Mr. Larry Cooper	(07) 3824 9541	Department of Primary Industries and Fisheries (Qld)
UT-38A	Rejuvenation of peppermint crops through establishment of quality nursery stock	Prof. Robert Menary	(03) 6226 2723	University of Tasmania
UT-40A	Improved oil quality and yield from Tasmanian-grown French lavender	Prof. Robert Menary	(03) 6226 2723	University of Tasmania
UT-49A	Increased yields from peppermint crops through improved micronutrient nutrition	Prof. Robert Menary	(03) 6226 2723	University of Tasmania
ISO-1A	Production of newsletter of the Essential Oil Producers Association of Australia	Dr Ian Southwell	(02) 6624 2453	Ian Southwell

Regulatory Approvals

UT-45A

Quality assurance protocols to minimise pesticide residues in essential oils

Prof. Robert Menary

(03) 6226 2723

University of Tasmania

Development and Improvement of Products

Project Title Development and economic assessment of Artemisia for production of anti-malarial extract

RIRDC Project No.: BRA-4A

Start Date: 01-Jan-06

Finish Date: 31-Dec-09

Researcher: Dr Rowland Laurence

Organisation: Botanical Resources Australia
44-46 Industrial Drive
ULVERSTONE TAS 7315

Phone: (03) 6425 5888

Fax: (03) 6425 4468

Email: tgroom@botanicalra.com.au

Objectives To develop commercially viable techniques for the production of marketable extracts of artemisia, through the investigation of ways to maximise the yield of artemisinin, the anti-malarial compound, present in Artemisia annua L.

Current Progress This project, funded from January 2006, will attempt to determine a production method for Artemisia annua, which will be economically viable in Australia. Key areas for investigation will be seed handling and sowing techniques, weed control and harvesting systems.
Seed of two improved lines of A. annua have been acquired to date and sowing in the field attempted, but with poor emergence of the small and fragile seed. Initial investigations of the plant’s tolerance of herbicides have also been attempted but with little result to date due to this poor emergence in the field.
Greenhouse-grown seedlings also were transplanted in the field in January and an initial assessment of their tolerance of some commercial herbicides was made. These results have been more encouraging and an attempt to verify this data will be made next season. The remainder of these transplants will be harvested shortly and artemisinin content determined.
These plants have not flowered in the Tasmanian environment to date and strategies to produce additional seed for experimentation are being considered.

Development and Improvement of Products

Project Title Analysis of Dodonea viscose as a potential remedy for anti-inflammatoryand anti-bacterial applications

RIRDC Project No.: UNC-21A

Start Date: 01-Jul-03

Finish Date: 31-Jul-07

Researcher: Mr Andrew Pengelly

Organisation: The University of Newcastle
Ourimbah Campus
Science Building, Room SOE1.23
Brush Road
OURIMBAH NSW 2258

Phone: (02) 4349 4490

Fax: (02) 4348 4145

Email: andrew.pengelly@newcastle.edu.au

Objectives ·1 To develop analytical techniques to analyse for saponins, flavonoids, coumarins and diterpenes found in Dodonaea viscose var. Angustifolia.
·2 To determine levels of each constituent group in various plant sections and determine the effects of some variables such as growing location
·3 To test extracts with a known biochemical matrix for antiinflammatory and antibacterial properties.

Current Progress Anti-oxidant capacity
The previous report (November 2005), presented results of peroxyl-trapping activity of Dodonaea viscosa leaf extracted with ethanol, aqueous-water and hot water. Subsequently these assays were replicated using more standardised concentrations and including extracts prepared with cold water. In order to add more statistical power to the results 10 replicates were performed.
With the exception of a 0.5% ethanol extract (mean inhibition 20%) all Dodonaea leaf extracts demonstrated levels of inhibition that were equivalent to or higher than TROLOX, the vitamin E analogue. Hot water extracts showed 95% inhibition at 4%, 92% inhibition at 2% and 79% inhibition at 1%. The other notable result came from the 4% ethanol extract, which produced a mean inhibition rate of 96%.
Fibroblast Proliferation assay
To assess the proliferative effects of Dodonaea extracts on human fibroblasts, cells grown to near confluence are combined with different concentrations of extracts.
Cell proliferation induces chemical reduction of the media, resulting in change of colour from blue to red, and is determined by measuring absorbance at 570nm and 600nm in a microplate reader.
Currently fibroblasts are being cultured and sub-cultured in preparation for this assay.

Improved Production Systems

Project Title Commercial production of milkweed, a plant with anti-cancer properties

RIRDC Project No.: DAQ-329A

Start Date: 01-Jul-05

Finish Date: 30-Nov-07

Researcher: Mr Larry Cooper

Organisation: Department of Primary Industries and Fisheries (Qld)
PO Box 327
CLEVELAND QLD 4163

Phone: (07) 3824 9541

Fax: (07) 3286 3094

Email: larry.cooper@dpi.qld.gov.au

Objectives Milkweed (Euphorbia peplus) has bioactives known as PEP compounds and these are suitable for use in treating various cancers, particularly skin cancer. This research will build on the system for commercially growing of E. peplus, which was developed as a result of previous projects funded by RIRDC. Cost effective production techniques will be enhanced to enable producers to meet market needs and enable the even and reliable production of PEP compounds all year round for a larger market. Efficient production systems for both intensive cropping in artificial structures and less intensive broad-acre field production will be developed. Research to date was focused on production of material for clinical trials. This research will result in the development of intensive production techniques for commercial scale production and extraction. The growing sector which will benefit from this research would be small holdings and holdings with some covered or protected areas such as shade or plastic tunnel. The growing E. peplus would aso be attractive to organic growers.

Current Progress Trial initiated in early March 2006 to look at plant spacing, planting method (plugs vs direct seeding), plastic mulch vs bare ground and fertilizers (amount of nitrogen). Harvesting commenced early May and is still in progress. Determination of PEP compound concentrations will occur after all harvesting is completed.
Seed storage testing and supply system to growers was started. Seed is stored at DPI&F Redlands Research Station and distributed, via Postpack, on request from Peplin. This system is working well to supply seed to plug and plant growers in Queensland and New South Wales. Germination testing is still to be fully explored and a commercial lab may be needed.
Meeting of researchers, growers and Peplin was held on 11^th April 2006 at Stanthorpe and was attended by 9 growers, 4 researchers and Steve Ogbourne from Peplin. Discussions and presentations focussed on light/shade, plugs/direct seed, use of pesticides and harvesting issues. Main outcomes were that growers preferred the use of plugs, plastic mulch (for weed control) and Peplin have now agreed to contract DPI&F staff to submit any minor use permit applications for pesticides. Growers networked and shared ideas freely.

Improved Production Systems

Project Title Rejuvenation of peppermint crops through establishment of quality nursery stock

RIRDC Project No.: UT-38A

Start Date: 18-May-03

Finish Date: 31-Jul-06

Researcher: Prof Robert Menary

Organisation: University of Tasmania
School of Agricultural Science
GPO Box 252-54
HOBART TAS 7001

Phone: (03) 6226 2723

Fax: (03) 6226 7609

Email: r.menary@utas.edu.au

Objectives The objective of the project is to establish a source of productive, clean peppermint material and the best method of propagation to achieve high yields. The project will deliver both an established nursery of clean, high yielding peppermint stock and a report to the industry on the best method for future production of such a nursery.

Current Progress Of the 200 clones tested in the first two years of this project, 51 were selected for further testing in a field trial. The trial includes a number of clones with potential rust resistance and a control group based upon a random selection of the original 200 mother plants. Samples are currently being steam distilled to assess oil content and oil composition. The data will be used to determine which clones should be further tested under commercial production conditions.
The propagation method trial showed that the plants were rejuvenated by both micropropagation and propagation by tip cuttings, resulting in plants with faster growth rates, higher oil yield and a shorter time to maturity and therefore harvest. Micropropagation did not provide evidence of sufficiently improved vigour to warrant further investigation of this method. Tip cuttings are a far more practical alternative and the results here suggest propagation of tip cuttings for the establishment of purpose-grown nursery sites should provide substantial benefits to growers.

Improved Production Systems

Project Title Improved oil quality and yield from Tasmanian-grown French lavender

RIRDC Project No.: UT-40A

Start Date: 01-Nov-03

Finish Date: 30-Nov-06

Researcher: Prof Robert Menary

Organisation: University of Tasmania
School of Agricultural Science
GPO Box 252-54
HOBART TAS 7001

Phone: (03) 6226 2723

Fax: (03) 6226 7609

Email: r.menary@utas.edu.au

Objectives The production of French lavender oil conforming to the ISO standard is possible only if suitable clones are harvested at the appropriate time. The primary objective of this proposal is to screen the available lavender germplasm for those clones producing oils of a desirable composition. These will be used to establish a nursery of elite mother stock from which material will be available for future industry expansion. Optimum harvest date for each clone will be determined by monitoring changes in oil composition through the season. The industry partner will conduct a field trial to determine the feasibility of a more intensive production system with increased nitrogen and irrigation inputs. The university will undertake the monitoring of oil yield and quality from this trial.

Current Progress A clone bank of 80 lavender clones is established and is available for multiplication and release to industry, as nursery stock, at the completion of the project. The multiplication phase will be based upon use of the improved micropropagation technique developed in the previous season.
Further field-grown flower samples were collected from each of these clones for further testing. These samples are currently being steam-distilled and will be available shortly.
Data assessing oil composition changes are now available for two seasons. These data indicate that harvest should be targeted to post-senescence of the corolla. At such a time there is a risk of yield loss due to floret abscission but examination of flowers in the field indicated that florets which set seed are tenacious and that the florets which fall are those which have not set seed. Future research should investigate the correlations between oil yield, oil composition and seed set in lavender.
Flower and herb samples have been collected from the intensive production system trial. These are being analysed for correlations between tissue nitrogen content, oil yield and oil composition.

Improved Production Systems

Project Title Increased yields from peppermint crops through improved micronutrient nutrition

RIRDC Project No.: UT-49A

Start Date: 01-Jul-05

Finish Date: 30-Nov-07

Researcher: Prof Robert Menary

Organisation: University of Tasmania
School of Agricultural Science
GPO Box 252-54
HOBART TAS 7001

Phone: (03) 6226 2723

Fax: (03) 6226 7609

Email: r.menary@utas.edu.au

Objectives Determine current micronutrient status of commercial fields and undertake diagnosis and recommendation for remedial action.

Current Progress A field trial was established to determine responses to S,Mo,Cu and B. Oil analysis of the first harvest has revealed that nutritional treatments have influenced composition of the oil. Menthone and menthyl acetate varied with treatment but menthol and menthofuran were not affected.The variation in oil composition is likely to be related to the physiological age of plants at harvest with, high levels of menthone in immature plants and high menthyl acetate in mature plants. A higher % oil yield was observed in treatments receiving B.
Glasshouse trials were conducted to establish critical levels in tissue for S,Mo,Cu and B. Dry weight per plant indicated that critical levels for growth were established for S, Mo and B. Levels of Cu in the soil were too high and deficiency levels were not recorded in these peppermint plants.
Work is continuing on the distillation and chemical analysis of the second oil harvest. Information from the first and second harvests will be combined to assess the overall effect of nutrition on growth oil yield and composition.

Improved Production Systems

Project Title Production of newsletter of the Essemtial Oil Producers Association of Australia

RIRDC Project No.: ISO-1A

Start Date: 02-Jan-06

Finish Date: 30-Jun-07

Researcher: Dr Ian Southwell

Organisation: 22 Canterbury Chase
GOONELLABAH NSW 2480

Phone: (02) 6624 2453

Email: southwells@optusnet.com.au

Objectives To inform growers, oil producers and traders of national and international developments in the trade of essential oils
Three issues of the newsletter will be published over a period of two years.

Current Progress Newsletter Issue 13 has been published and distributed. It contained information on: ·1 Forthcoming meetings
·2 Notes on recent conferences and symposia
·3 Regulatory matters
·4 Essential oils standards(Australian)
·5 Constitution of EOPAA
·6 Recently published articles and books
·7 RIRDC News and Publications

Regulatory Approvals

Project Title Quality assurance protocols to minimise pesticide residues in essential oils

RIRDC Project No.: UT-45A

Start Date: 01-Jun-04

Finish Date: 30-Aug-07

Researcher: Prof Robert Menary

Organisation: University of Tasmania

Phone: (03) 6226 2723

Fax: (03) 6226 7609

Email: r.menary@utas.edu.au

Objectives ·1 Develop a paper based on a chemical management plant for an approved supplier program
·2 Optimise analytical procedures for analysis of pesticides in essential oils
·3 Monitor pesticide behaviour in cropping cycle
·4 Undertake hazard analysis for pesticides used in crop production.

Current Progress The paper based chemical management plan and the HACCP based approved supplier program has been presented to our industry partners for assessment and implementation.
In response to industry requests, Select® (clethodim) , Eclipse® (metosulam) , Spinflo® (carbendazim), Asulox® (asulam) and dimethoate have been added to the list of potential targets for pesticide screening.
The implementation of routine solid phase extraction and cleanup steps to enhance pesticide detection levels remains elusive. This is primarily due to a wide range of chemical pesticides used in production and the complex matrices of essential oils.
Analysis of the field trials involving pesticide behaviour throughout the production cycle has been completed for peppermint, blackcurrant and fennel. The parsley trial is in the process of development and should be completed within the next month.
Spray boom calibration and maintenance procedures have been finalised and are ready to present to producers for assessment and implementation as part of an audited quality assurance programme.

Project Title	Development and economic assessment of Artemisia for production of anti-malarial extract
RIRDC Project No.:	BRA-4A
Start Date:	01-Jan-06
Finish Date:	31-Dec-09
Researcher:	Dr Rowland Laurence
Organisation:	Botanical Resources Australia 44-46 Industrial Drive ULVERSTONE TAS 7315
Phone:	(03) 6425 5888
Fax:	(03) 6425 4468
Email:	tgroom@botanicalra.com.au
Objectives	To develop commercially viable techniques for the production of marketable extracts of artemisia, through the investigation of ways to maximise the yield of artemisinin, the anti-malarial compound, present in Artemisia annua L.
Current Progress	This project, funded from January 2006, will attempt to determine a production method for Artemisia annua, which will be economically viable in Australia. Key areas for investigation will be seed handling and sowing techniques, weed control and harvesting systems. Seed of two improved lines of A. annua have been acquired to date and sowing in the field attempted, but with poor emergence of the small and fragile seed. Initial investigations of the plant’s tolerance of herbicides have also been attempted but with little result to date due to this poor emergence in the field. Greenhouse-grown seedlings also were transplanted in the field in January and an initial assessment of their tolerance of some commercial herbicides was made. These results have been more encouraging and an attempt to verify this data will be made next season. The remainder of these transplants will be harvested shortly and artemisinin content determined. These plants have not flowered in the Tasmanian environment to date and strategies to produce additional seed for experimentation are being considered.

Project Title	*Analysis of Dodonea viscose* as a potential remedy for anti-inflammatoryand anti-bacterial applications**
RIRDC Project No.:	UNC-21A
Start Date:	01-Jul-03
Finish Date:	31-Jul-07
Researcher:	Mr Andrew Pengelly
Organisation:	The University of Newcastle Ourimbah Campus Science Building, Room SOE1.23 Brush Road OURIMBAH NSW 2258
Phone:	(02) 4349 4490
Fax:	(02) 4348 4145
Email:	andrew.pengelly@newcastle.edu.au
Objectives	·1 To develop analytical techniques to analyse for saponins, flavonoids, coumarins and diterpenes found in Dodonaea viscose var. Angustifolia. ·2 To determine levels of each constituent group in various plant sections and determine the effects of some variables such as growing location ·3 To test extracts with a known biochemical matrix for antiinflammatory and antibacterial properties.
Current Progress	Anti-oxidant capacity The previous report (November 2005), presented results of peroxyl-trapping activity of Dodonaea viscosa leaf extracted with ethanol, aqueous-water and hot water. Subsequently these assays were replicated using more standardised concentrations and including extracts prepared with cold water. In order to add more statistical power to the results 10 replicates were performed. With the exception of a 0.5% ethanol extract (mean inhibition 20%) all Dodonaea leaf extracts demonstrated levels of inhibition that were equivalent to or higher than TROLOX, the vitamin E analogue. Hot water extracts showed 95% inhibition at 4%, 92% inhibition at 2% and 79% inhibition at 1%. The other notable result came from the 4% ethanol extract, which produced a mean inhibition rate of 96%. Fibroblast Proliferation assay To assess the proliferative effects of Dodonaea extracts on human fibroblasts, cells grown to near confluence are combined with different concentrations of extracts. Cell proliferation induces chemical reduction of the media, resulting in change of colour from blue to red, and is determined by measuring absorbance at 570nm and 600nm in a microplate reader. Currently fibroblasts are being cultured and sub-cultured in preparation for this assay.

Project Title	Commercial production of milkweed, a plant with anti-cancer properties
RIRDC Project No.:	DAQ-329A
Start Date:	01-Jul-05
Finish Date:	30-Nov-07
Researcher:	Mr Larry Cooper
Organisation:	Department of Primary Industries and Fisheries (Qld) PO Box 327 CLEVELAND QLD 4163
Phone:	(07) 3824 9541
Fax:	(07) 3286 3094
Email:	larry.cooper@dpi.qld.gov.au
Objectives	Milkweed (Euphorbia peplus) has bioactives known as PEP compounds and these are suitable for use in treating various cancers, particularly skin cancer. This research will build on the system for commercially growing of E. peplus, which was developed as a result of previous projects funded by RIRDC. Cost effective production techniques will be enhanced to enable producers to meet market needs and enable the even and reliable production of PEP compounds all year round for a larger market. Efficient production systems for both intensive cropping in artificial structures and less intensive broad-acre field production will be developed. Research to date was focused on production of material for clinical trials. This research will result in the development of intensive production techniques for commercial scale production and extraction. The growing sector which will benefit from this research would be small holdings and holdings with some covered or protected areas such as shade or plastic tunnel. The growing E. peplus would aso be attractive to organic growers.
Current Progress	Trial initiated in early March 2006 to look at plant spacing, planting method (plugs vs direct seeding), plastic mulch vs bare ground and fertilizers (amount of nitrogen). Harvesting commenced early May and is still in progress. Determination of PEP compound concentrations will occur after all harvesting is completed. Seed storage testing and supply system to growers was started. Seed is stored at DPI&F Redlands Research Station and distributed, via Postpack, on request from Peplin. This system is working well to supply seed to plug and plant growers in Queensland and New South Wales. Germination testing is still to be fully explored and a commercial lab may be needed. Meeting of researchers, growers and Peplin was held on 11^th April 2006 at Stanthorpe and was attended by 9 growers, 4 researchers and Steve Ogbourne from Peplin. Discussions and presentations focussed on light/shade, plugs/direct seed, use of pesticides and harvesting issues. Main outcomes were that growers preferred the use of plugs, plastic mulch (for weed control) and Peplin have now agreed to contract DPI&F staff to submit any minor use permit applications for pesticides. Growers networked and shared ideas freely.

Project Title	Rejuvenation of peppermint crops through establishment of quality nursery stock
RIRDC Project No.:	UT-38A
Start Date:	18-May-03
Finish Date:	31-Jul-06
Researcher:	Prof Robert Menary
Organisation:	University of Tasmania School of Agricultural Science GPO Box 252-54 HOBART TAS 7001
Phone:	(03) 6226 2723
Fax:	(03) 6226 7609
Email:	r.menary@utas.edu.au
Objectives	The objective of the project is to establish a source of productive, clean peppermint material and the best method of propagation to achieve high yields. The project will deliver both an established nursery of clean, high yielding peppermint stock and a report to the industry on the best method for future production of such a nursery.
Current Progress	Of the 200 clones tested in the first two years of this project, 51 were selected for further testing in a field trial. The trial includes a number of clones with potential rust resistance and a control group based upon a random selection of the original 200 mother plants. Samples are currently being steam distilled to assess oil content and oil composition. The data will be used to determine which clones should be further tested under commercial production conditions. The propagation method trial showed that the plants were rejuvenated by both micropropagation and propagation by tip cuttings, resulting in plants with faster growth rates, higher oil yield and a shorter time to maturity and therefore harvest. Micropropagation did not provide evidence of sufficiently improved vigour to warrant further investigation of this method. Tip cuttings are a far more practical alternative and the results here suggest propagation of tip cuttings for the establishment of purpose-grown nursery sites should provide substantial benefits to growers.

Project Title	Improved oil quality and yield from Tasmanian-grown French lavender
RIRDC Project No.:	UT-40A
Start Date:	01-Nov-03
Finish Date:	30-Nov-06
Researcher:	Prof Robert Menary
Organisation:	University of Tasmania School of Agricultural Science GPO Box 252-54 HOBART TAS 7001
Phone:	(03) 6226 2723
Fax:	(03) 6226 7609
Email:	r.menary@utas.edu.au
Objectives	The production of French lavender oil conforming to the ISO standard is possible only if suitable clones are harvested at the appropriate time. The primary objective of this proposal is to screen the available lavender germplasm for those clones producing oils of a desirable composition. These will be used to establish a nursery of elite mother stock from which material will be available for future industry expansion. Optimum harvest date for each clone will be determined by monitoring changes in oil composition through the season. The industry partner will conduct a field trial to determine the feasibility of a more intensive production system with increased nitrogen and irrigation inputs. The university will undertake the monitoring of oil yield and quality from this trial.
Current Progress	A clone bank of 80 lavender clones is established and is available for multiplication and release to industry, as nursery stock, at the completion of the project. The multiplication phase will be based upon use of the improved micropropagation technique developed in the previous season. Further field-grown flower samples were collected from each of these clones for further testing. These samples are currently being steam-distilled and will be available shortly. Data assessing oil composition changes are now available for two seasons. These data indicate that harvest should be targeted to post-senescence of the corolla. At such a time there is a risk of yield loss due to floret abscission but examination of flowers in the field indicated that florets which set seed are tenacious and that the florets which fall are those which have not set seed. Future research should investigate the correlations between oil yield, oil composition and seed set in lavender. Flower and herb samples have been collected from the intensive production system trial. These are being analysed for correlations between tissue nitrogen content, oil yield and oil composition.

Project Title	Increased yields from peppermint crops through improved micronutrient nutrition
RIRDC Project No.:	UT-49A
Start Date:	01-Jul-05
Finish Date:	30-Nov-07
Researcher:	Prof Robert Menary
Organisation:	University of Tasmania School of Agricultural Science GPO Box 252-54 HOBART TAS 7001
Phone:	(03) 6226 2723
Fax:	(03) 6226 7609
Email:	r.menary@utas.edu.au
Objectives	Determine current micronutrient status of commercial fields and undertake diagnosis and recommendation for remedial action.
Current Progress	A field trial was established to determine responses to S,Mo,Cu and B. Oil analysis of the first harvest has revealed that nutritional treatments have influenced composition of the oil. Menthone and menthyl acetate varied with treatment but menthol and menthofuran were not affected.The variation in oil composition is likely to be related to the physiological age of plants at harvest with, high levels of menthone in immature plants and high menthyl acetate in mature plants. A higher % oil yield was observed in treatments receiving B. Glasshouse trials were conducted to establish critical levels in tissue for S,Mo,Cu and B. Dry weight per plant indicated that critical levels for growth were established for S, Mo and B. Levels of Cu in the soil were too high and deficiency levels were not recorded in these peppermint plants. Work is continuing on the distillation and chemical analysis of the second oil harvest. Information from the first and second harvests will be combined to assess the overall effect of nutrition on growth oil yield and composition.

Project Title	Production of newsletter of the Essemtial Oil Producers Association of Australia
RIRDC Project No.:	ISO-1A
Start Date:	02-Jan-06
Finish Date:	30-Jun-07
Researcher:	Dr Ian Southwell
Organisation:	22 Canterbury Chase GOONELLABAH NSW 2480
Phone:	(02) 6624 2453
Email:	southwells@optusnet.com.au
Objectives	To inform growers, oil producers and traders of national and international developments in the trade of essential oils Three issues of the newsletter will be published over a period of two years.
Current Progress	Newsletter Issue 13 has been published and distributed. It contained information on: ·1 Forthcoming meetings ·2 Notes on recent conferences and symposia ·3 Regulatory matters ·4 Essential oils standards(Australian) ·5 Constitution of EOPAA ·6 Recently published articles and books ·7 RIRDC News and Publications

Project Title	Quality assurance protocols to minimise pesticide residues in essential oils
RIRDC Project No.:	UT-45A
Start Date:	01-Jun-04
Finish Date:	30-Aug-07
Researcher:	Prof Robert Menary
Organisation:	University of Tasmania
Phone:	(03) 6226 2723
Fax:	(03) 6226 7609
Email:	r.menary@utas.edu.au
Objectives	·1 Develop a paper based on a chemical management plant for an approved supplier program ·2 Optimise analytical procedures for analysis of pesticides in essential oils ·3 Monitor pesticide behaviour in cropping cycle ·4 Undertake hazard analysis for pesticides used in crop production.
Current Progress	The paper based chemical management plan and the HACCP based approved supplier program has been presented to our industry partners for assessment and implementation. In response to industry requests, Select® (clethodim) , Eclipse® (metosulam) , Spinflo® (carbendazim), Asulox® (asulam) and dimethoate have been added to the list of potential targets for pesticide screening. The implementation of routine solid phase extraction and cleanup steps to enhance pesticide detection levels remains elusive. This is primarily due to a wide range of chemical pesticides used in production and the complex matrices of essential oils. Analysis of the field trials involving pesticide behaviour throughout the production cycle has been completed for peppermint, blackcurrant and fennel. The parsley trial is in the process of development and should be completed within the next month. Spray boom calibration and maintenance procedures have been finalised and are ready to present to producers for assessment and implementation as part of an audited quality assurance programme.