Varietal development and disease management of Ixodia achillaeoides for cutflower production

A report for the Rural Industries Research and Development Corporation

Gail Barth and Barbara Hall South Australian Research and Development Institute

March 2001

RIRDC Publication No 00/186 RIRDC Project No DAS-45A

© 2001 Rural Industries Research and Development Corporation. All rights reserved.

ISBN 0 642 58217 3 ISSN 1440-6845

Varietal development and disease management of Ixodia achillaeoides for cutflower production Publication No. 00/186 Project No DAS-45A

The views expressed and the conclusions reached in this publication are those of the author and not necessarily those of persons consulted. RIRDC shall not be responsible in any way whatsoever to any person who relies in whole or in part on the contents of this report.

This publication is copyright. However, RIRDC encourages wide dissemination of its research, providing the Corporation is clearly acknowledged. For any other enquiries concerning reproduction, contact the Publications Manager on phone 02 6272 3186.

Researcher Contact Details Gail Barth South Australian Research and Development Institute 2b Hartley Grove URRBRAE SA 5064 GPO Box 397 ADELAIDE SA 5001

Phone: 08 8303 9400 Fax: 08 8303 9424 Email: [email protected]

RIRDC Contact Details Rural Industries Research and Development Corporation Level 1, AMA House 42 Macquarie Street BARTON ACT 2600 PO Box 4776 KINGSTON ACT 2604

Phone: 02 6272 4539 Fax: 02 6272 5877 Email: [email protected] Web: http://www.rirdc.gov.au

Published in March 2001 Printed on environmentally friendly paper by Canprint

ii Foreword

Ixodia achillaeoides is a wildflower crop which has been bush harvested for many years and cultivated increasingly over the past 20 years. The main objective of this program was to increase the production levels of Ixodia daisy for export markets by overcoming varietal and production based problems which limited its uptake and expansion. To achieve this objective a varietal improvement and disease management program were commenced with the aim of providing improved varieties of Ixodia to industry.

The varietal improvement program incorporated: the collection and preservation of varieties used by industry or in previous assessment programs (SARDI), the location and assessment of major native stands throughout SA and Victoria and collection of ‘type’ varieties, collection of variants and outstanding individual varieties in terms of appearance or vigour and a breeding program. By increasing the availability of varieties, this program aimed to: i. extend the harvest period, ii. provide new products for dried and fresh flower markets and iii. improve quality characteristics including stem length, floral appearance, post harvest life and disease resistance.

Diseases of ixodia required identification, pathogenicity testing, prioritising and development of control measures to assure that ixodia production can proceed as a profitable venture.

This report, a new addition to RIRDC’s diverse range of over 600 research publications, forms part of our Wildflowers and Native Plants R&D program which aims to improve the profitability, productivity and sustainability of the Australian wildflower and native plant industry.

Most of our publications are available for viewing, downloading or purchasing online through our website: • downloads at www.rirdc.gov.au/reports/Index.htm • purchases at www.rirdc.gov.au/eshop

Peter Core Managing Director Rural Industries Research and Development Corporation

iii Acknowledgments

This project was financially supported by the Rural Industries Research and Development Institute. The authors also wish to thank The Banksia Company of Furner, SA (Mary and Tony Whitehead) and Cutting Flowers and Foliage of Millicent, SA (Heather and David Brown) for in-kind support of this project and managing trial sites. Appreciation is extended to all growers who contributed varieties for assessment, especially to Shane and Leila Huebner, Philip Dowling and Kylie Treble who assisted us in the location of new ixodia varieties and for their advice.

The authors wish to especially acknowledge and thank the SARDI staff who participated in this project work through technical assistance: Shona Chinnock, Jan Niejalke, Midori Jones, Claudia Bonnielle, Robin McMahon, Roz Varcoe, Alex Freebairn, Samuel Burton, Kerryn O’Brien and Catherine Hitch.

Associate researchers also contributed to this project including: Dr Greg Walker for undertaking the experimental and identification work on the nematodes, Dr. Trevor Wicks, SARDI Horticultural Pathology and his colleagues who assisted with pathogen identification, Dr. Bob Chinnock of the SA State Herbarium for botanical advice and Debra Partington of Biometrics SA for assistance with statistical analysis. Chris Salter and Mark Bartetzko of Primary Industries, SA are also thanked for their participation in the establishment of trials and technology transfer programs.

iv Contents

Foreword...... iii Acknowledgments...... iv Executive Summary ...... vii 1. Collection and development of Ixodia varieties...... 1 1.1 Introduction...... 1 1.1.1 Description and geography of native stands...... 1 1.1.2 History of cultivation...... 1 1.1.3 Cultivation ...... 2 1.2 Materials and Methods ...... 2 1.2.1 New Field Collections ...... 2 1.2.2 Growers contributions...... 4 1.2.3 Seed evaluation...... 4 1.2.4 Hybridisation ...... 5 1.3 Results...... 5 1.3.1 Characteristics of geographic forms ...... 5 1.3.2 Characteristics of floral form ...... 7 1.3.3 Propagation ...... 8 2. Variety Assessment Program...... 10 2.1 Introduction...... 10 2.2 Materials and methods ...... 10 2.2.1 Yield and performance trials ...... 10 2.2.2 Post harvest assessments...... 11 2.2.3 Stem Growth Measurements ...... 12 2.2.4 Variety Descriptions and Identification...... 12 2.2.5 PBR Comparative Trials ...... 12 2.2.6 Flowering controls and pot plant assessment...... 13 2.2.7 Assessment of Landscape Varieties...... 13 2.3 Results...... 13 2.4 Yield and performance trials...... 14 2.4.1 Field Trials 1997/98 - Lenswood...... 15 2.4.2 Field Trials 1997/98 – Southeast ...... 19 2.4.3 1997/99 Harvest data comparisons – Lenswood...... 22 2.4.4 Extended trial – Varieties 60- 67...... 24 2.5 Post harvest – fresh flower evaluation...... 26 2.6 Stem growth measurements...... 30 2.7 Variety Descriptions and Identification ...... 32 2.7.1 Harvest date records...... 32 2.7.2 Floral descriptions...... 33 2.7.3 Colour data from all the assessed forms ...... 36 2.8 Assessment of dwarf plants for pot plant production...... 37 2.8.1 Length of coolstore treatment affects growth and flowering ...... 38 2.8.2 Coolstore treatment enhances compactness...... 40 2.8.3 Bonzi® treatments affect dwarfing...... 41 2.8.4 Discussion ...... 42 2.9 Landscape variety assessment...... 43

v 3. Diseases of Ixodia: identification, varietal assessment and evaluation of control methods ...... 46 3.1 Introduction...... 46 3.2 Pathogen identification...... 46 3.2.1 Materials and methods ...... 46 3.2.2 Results...... 47 3.3 Pathogenicity studies...... 49 3.3.1 Materials and methods ...... 50 3.3.2 Fungi...... 50 3.3.3 Results...... 50 3.4 Testing methods of inoculum for Phytophthora...... 51 3.4.1 Materials and methods ...... 51 3.4.2 Results...... 52 3.5 Cultivar susceptibility...... 52 3.5.1 Materials and methods ...... 53 3.5.2 Results and discussion...... 54 3.6 Nematode and Verticillium Disease complex ...... 58 3.6.1 Materials and methods ...... 58 3.6.2 Results and discussion...... 58 3.7 Control of Powdery mildew...... 59 3.7.1 Materials and methods ...... 59 3.7.2 Results and discussion...... 60 3.8 Control of Phytophthora...... 62 3.9 Greenhouse experiments ...... 62 3.9.1 Materials and methods ...... 62 3.9.2 Results and discussion...... 63 3.10 Field phytophthora inoculation trial...... 64 3.10.1 Materials and methods...... 65 3.10.2 Results and discussion...... 65 4. Commercialisation...... 68 4.1 Introduction...... 68 4.1.1 Background, relevance and potential benefits...... 68 4.1.2 Economic benefits...... 68 4.2 Research strategies and methodology...... 69 4.3 Communications/adoption/commercialisation strategy ...... 69 5. Appendices ...... 70 5.1 Appendix I.: Records of herbaria searches for locations of Ixodia ahillaeoides ssp.alata 1 collections...... 70 5.2 APPENDIX II: Summary of harvest dates of all Ixodia varieties assessed at the Lenswood site, grouped by geographic origin...... 75 6. References...... 77

vi Executive Summary

Ixodia achillaeoides is a wildflower crop which has been bush harvested for many years and cultivated increasingly over the past 20 years. Demand for dried Ixodia blooms is strong on both export and domestic markets. Ixodia is very diverse in its forms, appearance and geographic origins and there are many unassessed varieties and seedlings under cultivation. Production of varieties with unknown performance records has led to poor quality control of the product and lack of confidence by growers in cultivation practices. With increasing cultivation, a range of disease problems have emerged which have seriously affected or restricted production of ixodia. These diseases require identification, pathogenicity testing and control measures, to allow ixodia production to proceed as a profitable venture.

The main objective of this program was to increase the amount and the quality of Ixodia daisy produced for export markets by overcoming varietal and production based problems which have limited commercial uptake and expansion of this crop. To achieve this objective a varietal collection, assessment and development program was commenced to provide improved varieties of Ixodia to industry.

The varietal improvement program incorporated:

• collection and preservation of varieties used by industry (1995-6) or in previous assessment programs (SARDI) • location and assessment of major native stands of Ixodia throughout SA and Victoria and collection of ‘type’ varieties to represent the complete geographic array • search for variants and outstanding individual varieties in terms of appearance or vigour • a breeding program.

By increasing the availability of varieties, this program aimed to: i. extend the harvest period, ii. provide new products for dried and fresh flower markets and iii. improve quality characteristics including stem length, floral appearance, post harvest life and disease resistance.

Outcomes

Varietal collection, evaluation and improvement program

Sixty varieties from thirty-two populations and selections from contributions of collections from eleven growers were established in cultivation in replicated trials and evaluated for yield, stem characteristics, flowering and post harvest performance. New varieties from hybridisation and selection programs were established in subsequent years for further field testing.

The results of field performance trials of varieties from a wide range of geographic origins has demonstrated that outstanding cut flower forms originate from two distinct geographic regions in South Australia and Victoria. Varieties originating from Nelson and Adelaide Hills districts were found to be capable of producing the highest yielding plants, best adapted to cultivation conditions. Average total yield of all plants assessed at the main site was 1352 gr/plant (4.6 bunches) in the first year of harvest and 1341 (3.9 bunches) in the

vii second year. We suggest that varieties requiring more than an average of 20 stems to form a dried bunch are uneconomical to produce and should be replaced by varieties with 12-20 stems per bunch. New varieties with total yields of <1200 grams in their first year of harvest have been eliminated from the screening program. Yield in stem length classes was also determined for each assessed variety as well as flower and bunch characteristics, wastage and postharvest performance particularly to identify varieties with outstanding potential for fresh flower development.

Growth patterns of 5 varieties representing distinct geographic forms were monitored fortnightly over 18 months to establish differences in growth between forms and varieties which impact on management practices. Quality characteristics of cultivated ixodia have been defined which will allow for the continued development and selection of improved forms to support the economical production of this crop.

Results of post harvest assessment trials suggest that ixodia varieties suited for marketing as a fresh product require vase-life >10 days, stem lengths >40 cm and freedom from stem blackening in water. The average vase life of all the plants assessed in the program was 11.3 days with the longest lasting flowers in the range of 15-17 days. Geographic origin proved important, as forms designated Deep Creek and Mt Richmond performed significantly better in keeping-quality trials than all other forms. To date, the early Nelson varieties have been most frequently grown for fresh flower markets but our results indicate as a whole these have performed poorly in keeping quality.

viii Disease program

A disease survey conducted on ixodia in commercial plantings and native populations recovered 12 species of fungi, 12 nematodes, 2 bacteria and a phytoplasm. The pathogenicity of 7 species of fungi and 1 nematode were confirmed.

The most serious losses of Ixodia in commercial plantings have been from the disease Verticillium dahliae, which is more serious where levels of plant pathogenic nematodes are shown to be high. All 58 cultivars tested in this program were susceptible. There are no known controls for these diseases other than fumigation.

Phytophthora cinnamomi and P. cryptogea, serious diseases of most Australian native flower crops, cause characteristic decline and yellowing of ixodia plants prior to death. Susceptibility to these diseases varied among cultivars with significant differences in susceptibility relating to geographic classes. These results were used in selecting parent varieties in the breeding program.

The recommended treatment procedure resulting from our research for managing phytophthora in Ixodia is, firstly, isolating infections by removing plants and drenching the site and neighbouring plants with a fungicide such as Fongarid®. The remaining crop should then be treated using a regime combining applications of Ridomil Gold® or granules initially and maintenance sprays of Foli-R-Fos® as a preventative treatment. The planting area should then be rotated with a non-susceptible crop in the next planting cycle.

Leaf diseases including powdery mildew and botrytis were found to be increasingly a problem in crowded plantings where overhead irrigation was used or where there was a heavy weed problem. Nine treatments were evaluated for controlling powdery mildew and a wettable sulphur compound was found to be the most effective.

New products

Recent marketing experience suggests that both terminal and spray flowering forms of Ixodia are acceptable marketed as bunches of between 6 and 12 stems depending on stem thickness and number of flowers/stem. This is a distinct departure from the traditional densely packed terminal flower clusters generally marketed. Several selected varieties have been identified of distinct form to be marketed as new products.

Genetically dwarf varieties of Ixodia were selected which were suitable for production of flowering pot plants without the use of growth retardants. In our experimental work, material from eleven distinct geographic regions were assessed and outstanding dwarf forms were consistently sourced from one region in South Australia. Protocols for initiating flowers and scheduling flowering have been investigated for several varieties. Other varieties have been identified with potential for development as landscape plants.

The ongoing developmental program and results from the program have been widely publicised to industry through workshops, national and international conferences, seminars and publications. Key producers have participated in the program throughout the evaluation of varieties and disease surveys.

Recommendations on varieties for commercialisation will be made in a confidential report to RIRDC.

ix 1. Collection and development of Ixodia varieties

1.1 Introduction

1.1.1 Description and geography of native stands

Ixodia achillaeoides is a perennial shrub in the Asteraceae, which grows erect, up to 2 metres in its natural habitat. It is a highly variable species with several recognisable leaf forms which range from narrow and linear to broad-linear to lanceolate. Leaves are attached to the stems without petioles and often form pronounced wings down the stems (Copley, 1982). Flowers are arranged in terminal corymbs (indeterminate flat-topped or convex open inflorescences) or panicles (compound inflorescences) of varying size and density which are dependant on the age and health status of the plant as well as on genetic factors.

The species is classified into three subspecies, chiefly ssp. alata, which is the most suitable for growing as a cut flower crop in terms of plant size, number of flowers and stem length. Ixodia achillaeoides ssp arenicola is native to the coastal district near Mt Richmond, SA to Portland, Victoria. Plants have large thick leaves and form a rounded dense plant with the largest flowers (>15mm) of all ixodia forms. Ixodia achillaeoides ssp achillaeoides occurs in primarily coastal regions where its distribution often overlaps with populations of subspecies alata. Plants in this group are often dwarf and have potential for use as landscape specimens or flowering pot plants.

Ixodia achillaeoides ssp. alata grows most commonly in open stringybark forest communities in the high rainfall districts of the Mt. Lofty Ranges in South Australia and South-western Victoria. This group is also represented on skeletal calcareous or lateritic soils in isolated coastal districts and on Kangaroo Island S.A., where the full range of variation in the subspecies is represented. Ixodia is a prominent understory plant in the Adelaide Hills, Glenelg River and Grampians National Parks in Victoria after fire or disturbance clears away other competing understory plants. Ixodia has been known by the common name Fireweed in some districts because of its regeneration and dominance after fires.

1.1.2 History of cultivation

Ixodia has been bush harvested and sold as a dried flower from the Adelaide Hills for over 50 years. Some pickers moved yearly from Victoria to South Australia following the progression of harvest from November to February. Large quantities of bush harvested material were shipped to West Australian wildflower exporters and were sent primarily to European markets in a mix with other Australian ‘bush flowers’. Ixodia did not develop an identity of its own in this market. Domestically, ixodia has a strong market and market identity with dried flower wholesalers and craftspersons and the quality and pricing of the product varies markedly. Ixodia is known in Australia by the common names of Hills daisy, Mountain daisy, Fireweed and South Australian daisy, however, the names Ixodia and Ixodia daisy are gaining wider recognition as common names for the cultivated flower.

There has been considerable interest in growing cultivated Ixodia over the past 15 years in S.A., Victoria and Western Australia and production has increased together with the developing Australian Wildflower Industry. Some producers directly export to buyers overseas that they have contacted and ship their product green (no attempt made to preserve

1 post harvest life but stems remain pliable) or dried after a period of storage. The main market is still domestic and demand exceeds supply. The total value of the Australian wildflower exports in 1997/98 was 27 million (RIRDC) with the estimated value of ixodia production being $330-520,000 in the same year (Barth, unpublished data).

1.1.3 Cultivation

Ixodia is one of the few everlasting-type flowers that is grown on a perennial shrub rather than as a herbaceous plant. Under cultivation, most forms are productive over a three year period, with subsequent decline in vigour or harvestable stem length after this time.

Variation in the overall plant form in cultivated varieties ranges from upright and narrow (0.5 m) to wide spreading (1.5 m) and vase-shaped. Desirable forms for cultivation are varieties that are self-branching and densely foliaged that can support multiple flowering stems and retain the ability to branch low in the bush after pruning. Most varieties respond favourably to early pinching or pruning to help establish low branching and a fuller bush. Productive plants can reach1.5 to 1.5 m during their second year of harvest.

Although Ixodia has been most commonly marketed as a dried flower, there is considerable potential for its development as a fresh cut flower crop. Special cultivation techniques and selected varieties are needed to produce high quality long stemmed fresh flowers. Early attempts in South Australia have shown good market acceptance of fresh ixodia blooms in Japan. Flowering stems are tolerant of packing and transport conditions and vase-life is adequate on all major forms. The temporary closure of the flower heads on contact with moisture or in conditions of high humidity is a drawback when auction systems are used in marketing. Some stem blackening has occurred in transport which is linked to cold moist storage, poor varietal selection or poor post harvest handling.

1.2 Materials and Methods

1.2.1 New Field Collections

At the start of this study, actively growing populations of ixodia were located with the assistance of local native plant enthusiasts, farmers, government authorities and records of the South Australian and Victorian Herbaria. Some isolated populations visited six years previously, when our department first began research on ixodia, have disappeared. New stands were located through fire records or where farming operations have disturbed native bush areas.

Typically, in an isolated stand, hundreds of individual seedlings were present and collection involved location of type specimens as well as selection of forms with unique characteristics. Representatives from the full clinal range of the species (including all three subspecies) were collected for our disease screening program.

All material collected from the wild was stored in coolers, transported and propagated by stem cuttings. Varieties were designated with accession numbers based on geographic origin and mother block plantings were established in the field (Lenswood Research Centre) with duplicate collections established in 200 mm pots and held in greenhouses and shadehouses at the Plant Research Centre, Adelaide. Stock plant collections were re-propagated every 6 months. The suitability of individual varieties to row culture was determined prior to their inclusion in the main assessment program.

2 Collecting trips in the first year of the research program were made to native stands of Ixodia in the following areas: Adelaide Hills (Forest Range, Lenswood, Balhannah, Deep Creek Conservation Park), Kangaroo Island (primarily Penneshaw and D’Estrees Bay areas), Grampians National Park, Black Ranges State Park and the lower SE of SA and Victoria (Nelson, Victoria, Glenelg River National Park in Victoria and Port McDonnell, SA). Outstanding individual varieties from these collections were included in the main field assessment program commenced in 1996.

Further collection trips were conducted in the second year to those sites were promising populations had been noted from previous visits. Collection permits were obtained from National Parks authorities and herbarium specimens were lodged with the South Australian and Victorian State Herbaria. Notes and GPS positioning were used to describe distinct populations which were not well represented in herbaria and which demonstrated unique characteristics in form. Flowering plants were bagged in the field for seed collection at a later date. New sites included: Pt Lincoln National Park, Anglesea, Victoria, a late blooming site in the Lower Glenelg River National Park in Victoria, Cape du Couedic on Kangaroo Island and more sites in the Adelaide Hills.

i ii

iii iv

Figure 1. Natural populations of Ixodia achillaeoides ssp alata at the extent of its range in: i. the Adelaide Hills in stringbark forest ii. coastal dunes in Port Lincoln National Park on Eyre Peninsula, SA, iii. on coastal cliffs at Anglesea, Victoria and iv. on sand plains in the Grampians National Park.

3 1.2.2 Growers contributions

Contributions of selected forms from growers and propagators were actively sought in the first and second years of the program for inclusion for the field assessment trials. Research agreements were provided to all contributors which protected varieties from commercialisation and provided confidentiality of results during the course of the assessment program. Records were documented of the unique features of each contributed variety to assist in limiting duplications and to avoid replication of large numbers of similar forms. Several contributed varieties were eliminated from the field trials but all questionable material was held in the stock plant collection for further evaluation if required.

Two notable collections were added at the end of the first year of our program, where the growers had themselves conducted evaluation programs over a number of years. In one case, we were only able to propagate one variety from a range of cutting material that was sent to us. Subsequently we visited the site and noted that the age and disease status of the plants made any future propagation impossible and this collection was lost.

Additionally, two varieties from another contribution that appeared very promising were eliminated when they tested positive to plant phytoplasms which were originally suspected after observations of growth of the first generation stock plants. We revisited the source plantings to attempt recollecting clean material, however all subsequent cuttings also tested positive for the disease.

Collections held by SARDI at the commencement of this project included geographic forms and selected varieties from sites in the Adelaide Hills, lower SE, Kangaroo Island and hybrids of geographic forms. These varieties were designated 32-40 and 47 in the main field assessment trial.

1.2.3 Seed evaluation

A third source of potential new varieties was from seed collected from field populations. Seed was collected for evaluation from populations where: i. plants were too old or in poor condition to evaluate, ii. where the population had declined and there were few remaining individuals (e.g. Deep Creek Conservation Park) or iii. where the population demonstrated extreme variability in form of flowers and vegetative traits and there was potential for selecting promising new variants.

Ixodia seed has an after-ripening requirement and may take from 3-6 months after harvest before some germination will occur. Germination was enhanced with heat, gibberellin and liquid smoke treatments. Seedlings were grown on and screened in row culture and in pots for further evaluation and promising individuals (<10%) were propagated for field evaluation or utilised in a crossing program. Off-season flowering of seedlings by manipulating flowering of tube stock with temperature and daylength controls was used to speed up the evaluation process.

4 1.2.4 Hybridisation

With the availability of a wide range of forms from throughout the geographic range of ixodia together in one place in our collections, we commenced a crossing program with the aim of finding intermediate characteristics between widely diverging forms. With this end in mind we are also utilising growth chambers to understand the control of flowering. The ability to control flowering allowed for the crossing of early and late flowering varieties which can not be achieved in the field, as well as assisting in programming pot plants for greenhouse production. (Barth et al, 1999a, Weiss et al ,1996.)

It may prove desirable in the future to chemically distinguish between forms to maintain cultivar integrity and identify cultivars to geographic origin or hybrid sources. Initial isozyme screening conducted by Dr Andrew Granger has shown that the high level of gums and resins in ixodia foliage interferes with electrophoretic analysis. It may be possible to utilise root tissue or separation techniques to allow for foliar analysis, however, these possibilities were not investigated further during the course of this program.

1.3 Results

1.3.1 Characteristics of geographic forms

In our experience, native stands of ixodia appearing after disturbance or fire have a life span of approximately 7 years. Shoot extension growth in years 5-6 is generally less than 5-10 cm, with the base of the plant becoming woody and the majority of the stems brown. Without soil disturbance or fire, populations of ixodia can exist only in the seed bank for many years. Herbarium records point to extensive populations of ixodia which are no longer in evidence due to competition from mature canopies, hand clearing or farming practices.

Stands of ixodia on Kangaroo Island are currently restricted to farming properties which have cleared native vegetation recently by fire or rolling to prepare grazing land. Ixodia can be a dominant plant in a newly disturbed site, and variation in local population characteristics is particularly noticeable in KI.

Deep Creek Conservation Park, which had a large population of Ixodia in 1991, when our department first started collecting forms, had a remnant population of less than 30 plants in 1995. Rooting success of cuttings taken from old or stressed plants is poor, and in the case of the population at Deep Creek, we only succeeded in rooting cuttings from one plant. Seed was collected from this population and after initial flowering and screening in pots, selections were added to the evaluation program. Varieties from this area are very interesting as they appear to be intermediate in many characteristics between Adelaide Hills forms and those from Kangaroo Island. Most importantly, they flower at an intermediate time (late December) which is important in adding continuity to harvest and for possible supply of fresh flowers during an important market period.

The disturbed margins of country fire service tracks in the Adelaide Hills have proven to be a good source of small populations of Ixodia. Collection trips in conservation parks and private land where there have been fires in the past three years also yielded many interesting variants which were selected for evaluation. The construction of a new national highway through a large native population of ixodia caused premature flowering of plants lining the banks of the

5 road, where high levels of exhaust fumes in confined roadways produced an ethylene effect and advanced flowering 8-10 weeks.

Populations of ixodia in the Grampians National Park Victoria are large and diverse throughout their entire range from Mt Zero in the north through to the southern Grampians. Some very distinct small flowered (<5mm diameter) populations occur. Controlled burns are part of the park management program, which assures constant regeneration of stands. In general, bushes are smaller (total height x width) than Adelaide Hills or Nelson forms, and stems and leaves are finer. Some of the most attractive flower forms come from these populations. Because large populations existed at the time of our visits, we were able to screen thousands of seedlings per day for promising variants such as doubled flower forms or extremely vigorous plants.

Table 1.1 provides a summary of some characteristics common to varieties originating from 11 distinct geographic areas in SA and Victoria.

Table 1.1 Summary of attributes of major geographic forms of Ixodia achillaeoides. ORIGIN OTHER BLOOM FLOWER OTHER CHARACTERISTICS NAMES PERIOD SIZE Central Mt. Adelaide SA Jan-Feb 8-10 mm Thick winged stems, tall bushes, Lofty Hills quality blooms, flat terminal Ranges clusters or sprays Lower Mt Deep Creek SA Dec 10-12 mm Intermediate in form and Lofty blooming period. Often well Ranges branched bush Kangaroo Is. SA Jan-Feb 6-8mm., Variable forms: fine foliaged >12mm small to medium flowers, dwarfs South-East SA Nov 12-15 mm Early, high yield first year, short- lived Yorke/Eyre SA Dec-Jan 5-7 mm Fine stems and foliage, small Peninsula bushes or dwarfs: potential for pot plants and landscape use, Nelson Glenelg Vic Nov-Dec 12-15 mm Variable, open sprays and flat River NP topped, Medium stems, high yielding, good quality Donovans SA Jan-Feb 12-15 mm Late form, similar to Nelson varieties Portland/ Vic Dec >15mm Largest flowers, wide leaves and Mt. thick stems. Short stem length Richmond Grampians Grampians Vic Feb-Mar 5-10mm Variable petal forms, small to NP med bushes, fine stems Attractive flower forms Lower Vic Feb-Mar 3-8 mm Smallest flower forms, distinct Grampians thin stems and leaves NP Anglesea, Vic Feb 8-10mm Pink buds, potential colour Vic variants

6 1.3.2 Characteristics of floral form

Variation in capitulum (flower) form can be very distinct within a population and between geographic forms but has not been one of the main criteria for selection of varieties for cultivation. Flower size and form on an individual plant remains constant but can be highly variable between individuals in a population. Flower diameter ranges from 3-15 mm throughout the subspecies (Figure 1.2 i) and papery ray petals can be arranged in single or double rows with petal form varying from elongated to broad with entire, notched or wavy margins. In some forms, the disc flowers are replaced by petaloids (Fig. 1.2 ii) which gives the appearance of a white centre. Some sterile mutants occur which are fully doubled and the capitula consists entirely of papery bracts.

i iii

ii

Figure 1.2 Examples of dried ixodia flowers and bunches: i. capitula from 4 varieties ranging in diameter from 3-12 mm, ii. clusters of dried flowers arranged in corymbs and showing differences in centre colour and iii. 50 and 60 cm bunches of terminally flowered stems.

The inflorescence of ixodia is usually terminal and composed of 3-80 capitula arranged in a corymb. The inflorescence itself can vary in shape from relatively flat to domed with the flowers densely crowded to open in arrangement. The size and shape of the inflorescence is a key criteria in varietal selection for cut flower production.

Floral characteristics which have importance in the floricultural development of this crop include the overall size of the inflorescence, showiness of the flowers, and visibility of the flowers in the inflorescences.

7 The appearance of dried bunches is greatly influenced by the shape of the inflorescence as well as the size, appearance and arrangement of the flowers. Some forms produce a flowering stem shaped as a spray with the development of secondary (smaller) lateral inflorescences subtending the terminal. This is more common during the first cropping season on vigorously growing plants and is also related to daylength conditions during flower initiation (Weiss et al., 1996). A high number of secondary inflorescences on a stem may require an additional hand stripping operation during bunching. Although these short-stemmed flowers have potential use in craft work and pot pourri, in reality they are difficult to market and present poor returns to the grower.

1.3.3 Propagation

Propagation of ixodia plants by stem cuttings has been achieved year-round on most varieties when stock plants are held in greenhouse or shade house conditions. Rooting will take place in 3-4 weeks on a mist bed with bottom heat (24oC). Some varieties appeared to be sensitive to IBA rooting compounds, particularly those formulated in 50% ethanol. Comparison of three rates and two formulations of IBA compared to non-treated controls in two varieties is summarised in Table 1.2. In this trial, cuttings were prepared in July and rooted for a period of 5 weeks. There are 20 replicates of each variety in each of the 5 treatments.

With variety 32, the highest rooting results were achieved with 3000ppm IBA in the Clonex formulation. Shoot damage was evident in all IBA treatments formulated with ethanol which would lead to the recommendation that cuttings remain untreated or that an IBA gel at 3000 ppm be used for highest rooting. With variety 283, the lowest rooting was achieved with Clonex 3000 ppm IBA formulation, where 75% of the cuttings were damaged and only 55% rooted in total. There is no significant difference between the control and the 1000 ppm IBA formulation which both gave 100% rooting. There is depressed rooting from all other formulations.

Varietal differences in these results may be caused by differences in growth flush patterns between varieties and relative lignification or ‘woodiness’ of the cuttings at any particular time cuttings are taken. Because of the evidence of sensitivity to ethanol preparations, talcs or gels at low concentrations would be recommended for any varieties that regularly gave less than 75% rooting when untreated with any rooting compounds.

8 Table 1.2. Rooting results and root ranking for two varieties of ixodia collected from greenhouse grown stock plants.

Damaged Damaged Dead or no Minor Good No damage No damage Variety Treatment rooting rooting rooting Good rooting Best rooting % Rooting 32 Control 4 13+ 3 80% 283 Control 12+ 6 100% 1,000 ppm 32 5 4 5 6 75% IBA 1,000 ppm 283 2 12 6 100% IBA 3,000 ppm 32 1 11 8 100% IBA clonex 3,000 ppm 283 9 6 1 4 55% IBA clonex 3,000 ppm 32 5 3 7 5 75% IBA 3,000 ppm 283 4 4 8 4 80% IBA 6,000 ppm 32 7 5 4 6 65% IBA 6,000 ppm 283 5 7 3 5* 75% IBA

Cuttings removed from mist beds at 5 weeks can be transplanted into 100 cm tubes at this stage or alternatively they can be direct rooted into speedling trays and hardened for a period of 3-5 weeks prior to transplanting into rows in the field.

When cuttings are collected from field grown plants, rooting is most successful on new season growth flushes that have hardened. Cuttings taken from varieties from the Nelson region can produce blind shoots unless terminal cuttings are used. Lateral bud development is strongly depressed by the terminals in these varieties and stem extension of up to 30 cm can occur without axillary buds. Propagation of all varieties during the period April-May has the greatest potential to lead to blind shoots and poor growth and would not be recommended, particularly for Nelson varieties

Cuttings taken from plants growing in the wild can be difficult to root if the plants are stressed from environmental conditions or age. It is often necessary to collect cuttings at different times of the year to establish the optimal growth flush stage for a particular variety or site.

9 2. Variety Assessment Program

2.1 Introduction

The aims of this program are to develop a range of suitable cultivars that: i. exhibit superior yield and quality, ii. extend the harvest period and iii. produce distinct products for dried, fresh and pot plant markets.

Field trials of selected varieties grown in randomised block plantings in South Australia were used to evaluate current varieties being commercially produced throughout Australia and to develop standards against which new cultivars of ixodia daisy can be compared. Criteria for assessment of suitable varieties include: total harvest yield (by weight and stem numbers in three length grades and number of dried bunches), number of stems/marketable bunch, harvest sequence, adaptability to cultivation and disease tolerance. Agronomic and post harvest performance as well as floral descriptions were used for selection of superior varieties for floriculture markets. As a result of a breeding program, 175 new varieties of ixodia have undergone field, greenhouse and post harvest assessments for development of superior cut flower forms.

2.2 Materials and methods

Stock plants of all varieties were assembled in 1995-96 into collections held in duplicate in shadehouses and greenhouses at the Plant Research Centre of SARDI, Urrbrae, SA. Stock plant mother blocks of selected varieties were established at Lenswood Centre in double row field plantings in the ornamental horticulture assessment block in the same years. All propagation material was taken from established plants held in our collections where identities from contributions were verified and replicates eliminated.

2.2.1 Yield and performance trials

Twenty cuttings each of fifty selected varieties were made the week of July 22-26, 1996 and placed under mist for a rooting period of 5 weeks. Plants were planted into 10mm tubes, hardened in a greenhouse, shadehouse and then outdoor holding area prior to field planting in October.

The main varietal assessment trials were established in October, 1996 at Lenswood Horticultural Centre (138°48'30" E; 34°58' S), elevation 600m, annual rainfall 1218 mm, in the Mount Lofty Ranges, SA and at The Banksia Company, Furner, SA (140° 14' E; 37° 21' S), Furner, SA in the lower SE district of the state on sandy soils.

Plants were placed in six randomised blocks spaced over 7 rows, replicated down the sloping site at each location. Plants were spaced at 1 x 2.5 metres to avoid interactions. Pre-plant applications of lime and single superphosphate were used (200 kg/ha rate applied in 1 metre planting strips only) and nitrogen and potassium applied at rates of 200 and 150 kg/ha in split applications, Oct. and Dec. as recommended from previous work at Lenswood centre (Maier et al 1994). These nutrients were applied as side-dressed fertilisers utilising ammonium nitrate and potassium sulphate granules. Nutricote controlled- release granules (equal mixture of 5-6 month and 12 month formulations) were mixed into the planting holes at rates of 5 gr/plant. Irrigation was applied by 4 l/hr pressure compensated drippers at rates of 8-12 l/plant, one to

10 two times per week during the period Oct-May at Lenswood and at the grower’s discretion at the South-east site. The pre-emergent herbicide Yield® was applied after planting and follow up sprays of glyphosate were applied with a controlled droplet applicator as needed.

Measurement of height and width of each plant at both trial sites was made in April, 1997 after 6 months of growth. There were no plant losses at this time.

The first harvest began in Oct. 1997 (early varieties) and extended to March, 1998. Peak harvest period for individual varieties is approximately 3-5 days, when flowers are fully opened and prior to peak pollen shed. In periods of extreme heat, peak harvest time was accelerated.

Harvest of each plant consisted of firstly collecting 3 stems for post-harvest analysis, tagging, placing in water and removing to a cool shed. The remainder of stems were cut at lengths of either 40 or 30 cm and formed into bunches as we worked. All replicates from one variety were harvested sequentially unless maturity on an individual plant varied (rare) and then harvest would be delayed. Bunches from each plant were individually bagged by stem class and the plant was then pruned to shape the bush and remove all damaged stems and undersized blooms. The prunings from each bush were separately bagged for calculation of waste weight. Data on total weights of stem classes and prunings were measured in the field and stem numbers and bunch weights measured in the shed. Bunches were then hung to dry for a period of 1-2 weeks when dry weight measurements were made. During extreme heat period, stems were cut to length in the field but weighed, graded and bunched in a shed to reduce moisture loss.

In the second year of harvest, stem classes were revised to 40 cm and >50 cm to more accurately reflect changing market demand for longer stems. The second harvest began (3/11/98) and was complete in March 1998.

In October 1998, an additional 8 varieties (60 –70) were planted in six randomised plots at the top of the previous trial (incorporated into the same irrigation system).

Also in Oct. 1998, leading varieties from the first year of assessment were established in a production trial which utilised close spacing and flower mesh to protect against plant damage and loss due to wind. 6 or 12 plants of each variety were planted in a double row, spaced alternately .6 m x .6 m and plus or minus support from 2 exterior wires running the length or the row, connected with internal cross wires or bamboo canes between plants. In adjacent rows, 32 new hybrids and field collections were established for evaluation. In October, 1999 additional varieties were established in field blocks.

2.2.2 Post harvest assessments

Three flowering stems were harvested at anthesis from each plant in the field assessment trial in 1997-98. This provided 18 stems of each variety for assessment where all replicates survived. One-year old replicated stockplants of newer varieties were also assessed in the same season. In total, 54 varieties were assessed.

11 At harvest, stems were cut to a length of 400 mm, labelled and immediately placed in buckets of water in the field then removed to a cool shed. Plants were transported within 6 hours of harvest to controlled environment rooms at the Plant Research Centre for vase-life assessment. Stems were assessed in a simple post harvest solution (250 ppm citric acid plus 200 ppm chlorine in deionised water) under continuous light in either a laboratory or coolstore kept at 20-22oC.

End of vase-life was determined by daily observation of leaf and stem decline. Stems and leaves fade quickly from an intense dark green to a dull green indicating stem plugging, or in some varieties the stem portion in water blackens prior to stem drying. Recordings were made of visual observations of blackening, leaf spotting or other evidences of decline in quality. Other quality considerations in post harvest life include: slow maturation of flowers (reduced pollen shedding), colour retention in the flower centres and reduced stem stickiness.

In the 1999/2000 harvest, 34 varieties were assessed, some replicating earlier results. Vase-life data were analysed by analysis of variance with least significant differences tested at P<0.05.

2.2.3 Stem Growth Measurements

Starting in August, 1997, fortnightly measurements were made of 12 marked stems (4 reps x 3 plants) of each of 5 varieties grown as part of the replicated varietal assessment trial at the Lenswood site. These measurements continued over a period of 18 months until January, 1999. New stems were marked on the same bushes after the first harvest and stem growth was plotted as incremental growth over the two harvest-year periods. The five varieties chosen represented three important geographic ecotypes of ixodia: Nelson (var 25), Adelaide Hills (var. 41) and the Grampians (var 49). In addition, var. 43, the earliest flowering commercial variety and var. 32, a hybrid with intermediate flowering dates were used for comparison.

2.2.4 Variety Descriptions and Identification

Flowering dates for all varieties over the 4 years of harvest were recorded at the Lenswood and South-east evaluation site and at commercial properties assessing some varieties. Plants were assessed two or three times per week in the flowering season to determine optimum harvest date, however a potential error of 2-3 days in any record is acknowledged. Floral measurements of all assessed varieties were made utilising dried bunches. Ten replicate stems were used and measurements made of flower diameter and flower centre diameter using a calliper, number of petals and colour of the flower centre (RHS colour guide).

2.2.5 PBR Comparative Trials

It is considered advantageous to protect some varieties of ixodia by plant breeders rights, particularly varieties developed in this program which may be marketed overseas. To determine the most suitable characteristics to use in comparative trials and to demonstrate varietal uniqueness, uniformity and stability, 10 plants each of 4 varieties were planted in October 1998 in a separate block at Lenswwod centre for data collection during their first harvest period in December – February 1999. Four varieties were used: one commercially known variety, two selections from our development program and one of hybrid origin. At harvest, measurements were made of a range of physical characteristics of stem and floral parts to use in variety comparisons.

12 2.2.6 Flowering controls and pot plant assessment It was desirable in our breeding program to manipulate flowering of ixodia plants to enable crossing of important Nelson and Adelaide Hills varieties which naturally flower four months apart. Through a combination of SD treatments and low temperatures, we were able to schedule flowering in different geographic forms to synchronise bloom periods and allowing crossing of varieties that could not be achieved in natural populations. For this work we utilised growth rooms or modified coolstores, with high intensity lighting and ebb and flow irrigation systems. Plants were treated in 150 mm pots for periods of 4-10 weeks for floral initiation and then removed to greenhouses or shadehouses for flower bud development. The length of flower initiation and development stage differed between varieties, however, the 8 week treatment was sufficient for flower initiation in all varieties.

In the development of flowering pot plants, flowering controls and growth retardants are both important in manipulating size and flower numbers. After screening a range of growth retardants, the formulation of pacrobutrazol, registered as Bonzi®, was found to be most effective in a single application in dwarfing ixodia. Three summary trials are presented in the results section which provide examples of the screening that was used to establish protocols for production of flowering pot plants with specific varieties.

2.2.7 Assessment of Landscape Varieties

Nine varieties were established in a field block for assessment as potential landscape selections. These included 7 naturally dwarf growing varieties identified from our collections in 1996-98 a variety held in greenhouse collections and one variety purchased from a nursery as a comparator. The latter was subsequently identified as a Kangaroo Island form after the wholesale nursery it originated from was traced. Varieties were planted in October, 1998 in blocks. 6 replicates planted .8 m apart in the row, 3 rows 2.5 m apart. Plant height and width measurements at bloom period, one year from planting were used to rank dwarfness (height) and size (h x w). Plant floriferousness, longevity and total impact will be used to assess landscape potential.

2.3 Results

Throughout the analysis and presentation of results of variety performance it was found useful to group varieties by classes of geographic origin (see Part 1, Table 1.1). This grouping also effectively sorts varieties by harvest date classes (early, mid and late) and allows for comparison of parameters which may have significance based on similarities arising from isolated populations. The following abbreviations are used throughout this section to refer to geographic classes:

SE = Southeast variety, origin unknown N = Nelson MR = Mt. Richmond DC = Deep Creek KI = Kangaroo Island E = Eyre/Yorke Peninsulas H = Adelaide Hills A = Anglesea G = Grampians

13 2.4 Yield and performance trials

Figure 2.1 Lenswood variety assessment trial at 6 months from planting in March 1996.

Measurements of plant height and width conducted six months after planting on all varieties gave an early indication of size differences between the two sites and significant differences in growth between varieties (Figure 2.1). The results of mean size measurements are presented in Figure 2.2, grouping varieties by their geographic origin which also separates them by harvest periods. The geographic classes in this figure are arranged with the earliest harvested varieties on the left (SE) in order to the latest flowering (G). The numbers in the centre of the bars indicates the number of varieties represented in the analysis. Each variety is represented by 12 plants.

Height x Width at 6 Months

4000

3500

3000

2500 ) 2

2000

HxW (mm 1500

1000 1161 33138 1161 3 3138

500

0 SE N MR DC KI H G SE N MR DC KI H G

South East Lenswood Variety

Figure 2.2 Comparison of average size (height x width) at six months of all varieites in field trials at two sites, grouped by geographic classifications.

14

The first most notable comparison that can be made between results at the two sites is that plants of the same age and variety growing at the South East site are on an average over 37% smaller than plants at the Lenswood site at six months of age. This is an early indication of site problems in the south-east although at this stage no plants had been lost. The variety designated SE is a commercial variety commonly used in this part of the state and well adapted to growing in the sandy soils if adequately irrigated. After six months of growth it is already 50% smaller that plants at Lenswood, indicating general problems, most likely insufficient irrigation.

At both sites, early flowering varieties (SE, N, DC) are significantly larger than later flowering varieties (H, G) which is consistant with their differing growth flushing characteristics (summarised in section 2). Error bars are presented for data summaries of each geographic region, showing that size variation among varieties is greatest at both sites in the KI forms. This is supported by field observations where great variability exists for a range of physical characteristics within most KI populations. The Adelaide Hills forms show less variation than Nelson or Grampian forms, again consistant with field observations.

The data in this table can be used to measure early performance of ixodia varieties or to assess young plantations. Comparisons can be made to the Lenswood data, which represent high quality, well managed plantings with known yield. The most accurate comparisons can be drawn between varieties of known geographic origin and the corresponding data presented here and for plantings established in spring after 6 months of growth.

2.4.1 Field Trials 1997/98 - Lenswood

Data collected from the two sites at first harvest in 1997/98 were analysed separately by general analysis of variance (AOV) using SAS, Version 8: SAS Institute Inc, Cary, N.C., USA. The results of the analysis from the Lenswood site across all variables is summarised in Table 2.1. with a listing of the p-values.

Table 2.1 Pr(F) values across all variables for data grouped as Class (early, mid or late season harvest), Geo (geographic class), Variety or Plant (rep).

15 Lenswood site Year 1 Analyses of variance across all variables Pr(F) d40w 40st 30w 30st was bu Tota f gt em gt em te lhar v C 2 0.04 0.00 0.29 0.00 0.90 0.55 0.18 l 016 058 999 000 393 171 345 a 39 00 52 03 47 35 58 s s G 3 0.00 0.37 0.00 0.00 0.34 0.00 0.00 e 023 424 344 014 578 000 000 o 61 75 47 40 39 01 06 V 3 0.00 0.00 0.00 0.00 0.00 0.00 0.00 a 2 000 000 001 000 000 000 000 r 02 01 51 02 07 00 00 i e t y P 5 0.82 0.63 0.77 0.98 0.96 0.89 0.89 l 962 131 981 140 076 034 905 a 08 80 28 19 03 22 48 n t

Tota 40b 30b Dry lma un un wgt ss 0.71 0.12 0.13 0.77 132 178 094 072 73 10 96 20 0.00 0.00 0.00 0.00 588 114 114 000 34 05 05 29 0.00 0.00 0.00 0.00 000 000 000 000 01 05 40 00 0.99 0.81 0.88 0.69 339 728 117 326 69 43 80 94

This analysis shows that variety is highly significant in all data categories, geographic class is significant for everything except for 40cm stem numbers and waste weight. Class is significant only for 40cm stem number and both classes of stem weight. Plant (rep) is not significant. The analysis of variance table for Total Harvest weight is presented in Table 2.2.

Table 2.2 Analysis of variance table of Total Harvest against the above classes as well as Loc. For Total Harvest: Df Sum of Sq Mean Sq F Value Pr(F) Class 2 619913 309957 1.88329 0.1565200 Loc 1 5986294 5986294 36.37259 0.0000000 Geo 2 140019 70010 0.42538 0.6544972 Variety 32 23686193 740194 4.4974 0.0000000 Residuals 121 19914489 164583

16 A plot of Geographic class versus total harvest shows that the locations KI and G are significantly different from the other Geo classes. The above analysis shows a two level factor “Loc” (1= DC, H AND N, 2= G and KI). After fitting this factor, Geo is not significant, which proves the locations DC, H and N are not significantly different and K and G are not significantly different. These results are illustrated in Figure 2.3.

Figure 2.3 Chart of mean total harvest weight of varieties harvested in Yr 1 at Lenswood, grouped by 5 geographic origin classes. Error bars are illustrated with dotted lines, grand means with solid circles and outlying values with hollow circles.

These results are important in demonstrating that plants originating from the two geographic areas, Nelson and the Adelaide Hills, from which most commercial production to date has been based, perform equally in the important assessment criteria of Total Harvest yield (grams). As these forms have very distinct harvest periods and vegetative and floral appearance, the use of varieties of both groups can be recommended with no required sacrifice of yield. Deep Creek, intermediary in harvest date between the two, also provides varieties with strong yield characteristics and can be an important contribution to providing continuity of harvest.

Kangaroo Island and the Grampians as geographic classes have significantly lower yield that the other groups, however, selected varieties deserved further assessment on yield characters as they approach the means of the N and H groupings.

The results of yield on a varietal basis (leading 18 varieties shown here) are presented in Table 2.3. Yield is reported as Total harvest weight in grams, weight in 2 stem length grades, bunch numbers, total biomass and dry weight. Total biomass includes total harvest weight plus all waste stems and prunings.

Table 2.3. Summary of the 1997/98 harvest results from 18 high yielding varieties at the Lenswood site. Varieties are listed from high to low in each data category, which present yield in terms of weight (grams) or bunch numbers.

17 Total harvest 40wgt 30wgt Bunch # Total Biomass Dry Weight variety mean variety mean variety mean variety mean variety mean variety mean 11 2488 11 1585 20 1150 40 8.9 17 4135 40 930 3 2240 23 1475 40 1077 11 8.7 11 3934 3 854 40 2100 17 1450 32 1028 48 7.0 48 3613 32 845 34 2045 5 1315 3 969 34 6.8 40 3471 17 820 48 2022 8 1283 11 888 10 6.5 37 3372 23 815 32 1995 34 1243 33 864 3 6.4 33 3365 11 814 36 1953 48 1240 7 838 32 6.1 34 3288 48 808 17 1950 37 1198 34 803 20 5.9 10 3140 36 798 5 1936 36 1191 10 755 37 5.8 32 3073 37 787 10 1923 31 1135 24 719 33 5.6 3 2992 34 785 24 1826 10 1125 48 695 17 5.5 50 2966 5 661 37 1800 22 1076 36 667 35 5.4 8 2895 10 661 23 1745 6 1063 16 573 36 5.4 24 2866 50 659 41 1601 24 1058 38 564 8 5.3 23 2765 24 656 44 1588 40 1023 39 563 5 5.2 20 2708 35 628 20 1529 3 1015 37 562 22 5.1 5 2705 6 621 33 1548 43 980 35 515 23 5.0 31 2663 8 579 8 1502 32 968 49 510 50 4.8 35 2604 30 564

All Var 1352 802 550 4.6 2416 545 LSD 473 399 307 1.75 503 200

The Total Harvest category, highlighted here, presents a summary of performance of leading varieties in comparison the to grand mean of 1352 grams for all varieties assessed at this site. The LSD for Total Harvest is 473, which groups the 5 top varieties (TH yields of 2488 - 2022 gr) as not significantly different from each other but with the top variety significantly different to all other varieties below variety 48 on this list. Looking across to the Bunch # category, four of the leading varieties are again represented with the addition of Variety 10. Varieties 11, 23 and 17 produced the highest weight of long stems, significantly different to all other varieties below variety 10 on the list.

The leading varieties are represented again at the top of the Dry Weight listing, with the addition of Variety 23 which has a high dry weight/fresh weight ratio (.467) which is desirable if bunches are being sold on a weight basis. Variety 11, by contrast, has the highest fresh weight and is ranked 6th in dry weight (ratio of .327). The relationship between TH (fresh) and dry weight for all harvested plants is shown in Figure 2.3 with an average ratio of dry weight/fresh weight at .400 for the entire data set. Outlying values (low ratio) represent varieties 25, 41, 33, 10 and 11.

18

Figure 2.3. Relation between total harvest weight of fresh stems and dry weight (grams) for all plants harvested in Year 1 at the Lenswood site.

2.4.2 Field Trials 1997/98 – Southeast

The results from the 1997/98 harvest at the Southeast site are summarised in Table 2.4. These results represent data from166 plants or 60% of the plants originally established in the trial. Plant losses in the period from the 6-month measurements until harvest (6-10 months later) were the result of wind damage and insufficient irrigation. The overall mean of Total Harvest is 745 grams per plant with a range of 2150g to 108g for the 48 varieties assessed. In terms of bunches, the grand mean for the site is 3.1 per plant with a range of variety yields of 8.0 to .5 per plant (LSD + 2.3). The top performing varieties are predominantly of Nelson origin, which may be an indication of site adaptability or more probably related to their early harvest dates and the fact that they would be less affected by insufficient irrigation. Stem length is reduced compared to production at the Lenswood site, with the majority of stems in the shorter, 30 cm class. Again, this is more pronounced in the Hills and Grampian varieties which have late harvest dates and are more reliant on stem elongation during the dry summer period. Variety 45, a Kangaroo Island form, is one of leading varieties at this site and one of the few that performed better here than at Lenswood. Due to the high mortality rate at this site, it was decided not to continue to manage these plants until a second harvest, but to re- establish an additional trial at a more protected site.

19 Table 2.4 Summary yield results of 1997/98 harvest at the Southeast site. Varieties are listed from high to low in each data category and only the top 20 varieties are presented.

Totalharv40wgt 30wgt bu 40stem 30stem Totstem Var mean Var mean Var mean Var meanVar mean Var mean Var mean 22 2150 3B 1430 22 1795 22 8.0 45 50 20 93 45 107 34 1733 9 1198 10 1132 20 7.0 9 48 22 87 22 93 3B 1600 34 1173 20 1023 34 6.2 3B 47 10 73 20 93 9 1537 45 932 32 940 32 6.2 23 41 32 71 18 85 32 1433 23 923 31 807 9 5.3 18 40 33 63 34 84 45 1400 15 498 40 791 45 5.0 15 37 19 61 9 83 23 1214 32 493 3 740 10 4.8 34 33 45 57 32 81 10 1132 24 440 25 693 3B 4.2 16 30 40 56 23 78 31 1063 16 435 37 628 40 4.1 24 27 49 55 49 75 20 1023 30 425 29 593 23 3.9 14 26 48 54 10 73 3 892 18 403 34 560 3 3.5 26 20 38 51 48 70 42 876 2 403 33 538 11 3.2 49 20 34 51 3B 68 40 791 14 400 21 532 31 3.2 30 19 44 48 15 65 25 785 22 355 38 498 48 3.2 2 18 37 47 16 63 21 773 11 350 48 497 33 3.2 42 16 21 46 33 63 48 753 42 334 43 486 42 3.1 48 16 18 45 19 61 2 751 26 283 45 468 21 3.1 11 14 11 44 14 60 16 742 31 257 42 457 14 3.0 21 13 42 42 21 59 11 739 48 256 35 431 2 3.0 47 13 29 40 11 58 Mean 745 290 453 3.1 14 40 54

Although the results at this site clearly reflect management practices that make varietal performance assessment difficult, there are some interesting comparisons that can be made between the two sites. Table 2.5 presents yield results at each site in terms of bunch numbers and the average number of stems in a bunch for each variety..

Yield in terms of commercial grade bunches (approx. 20cm diameter in 1990-97 markets) is reported here for the two sites to allow comparison of performance of individual varieties. The higher yielding varieties at the Lenswood site tended to yield less than 60% in terms of bunch numbers in the SE, with the exceptions of varieties 32 and 20.

20 Table 2.5 Comparison of yield of bunches and the number of stems in a bunch of all varieties assessed at both harvest sites in 1997/98.

Variety Lenswood South-east Combined Av bu/var st/bu bu/var st/bu bunches stems/bu 40 8.9 15.7 4.1 14.4 6.5 15.1 * 11 8.7 15.8 3.2 16.2 5.9 16.0 * 48 7 26.8 3.2 22.2 5.1 24.5 3B 6.8 11.9 4.2 15.7 5.5 13.8 * 34 6.8 16.3 6.2 13.2 6.5 14.7 * 10 6.3 15.0 4.8 14.7 5.5 14.8 * 32 6.1 26.7 6.2 11.5 6.1 19.1 20 5.9 12.1 7.0 15.4 6.5 13.8 * 37 5.8 19.1 2.8 17.2 4.3 18.2 33 5.6 24.1 3.2 18.4 4.4 21.2 17 5.5 17.6 2.5 17.9 4.0 17.8 * 36 5.4 16.6 1 14.0 3.2 15.3 35 5.4 19.7 3 19.0 4.2 19.4 5 5.1 16.2 1.8 16.7 3.5 16.5 * 3A 5 15.4 3.5 13.3 4.3 14.3 * 22 4.9 10.9 4.8 12.7 4.9 11.8 * 31 4.8 15.0 3.2 11.1 4.0 13.1 * 25 4.8 15.4 3.4 12.6 4.1 14.0 * 24 4.7 19.5 2.5 19.1 3.6 19.3 6 4.4 18.1 2.2 14.1 3.3 16.1 41 4.3 15.4 1.8 17.9 3.0 16.7 47 4.1 29.5 1.6 34.0 2.9 31.8 50 4.1 53.6 4.1 53.6 18 3.9 30.3 2.8 26.2 3.3 28.3 30 3.8 13.7 2.8 13.8 3.3 13.8 7 3.8 19.9 1.5 14.9 2.7 17.4 45 3.8 27.8 5.0 21.8 4.4 24.8 44 3.8 36.8 1.7 27.7 2.7 32.2 42 3.7 19.4 3.5 18.6 3.6 19.0 42 3.7 19.4 3.4 18.4 3.6 18.9 43 3.6 10.8 3.6 10.8 38 3.4 29.3 2.0 24.6 2.7 27.0 15 3.3 18.7 2.8 22.5 3.0 20.6 49 3.3 48.4 2.6 24.3 3.0 36.4 19 3.1 47.4 2.0 26.6 2.6 37.0 23 3 14.9 3.9 19.2 3.5 17.0 39 3 15.2 1.9 13.2 2.5 14.2 14 2.9 20.1 3.0 19.3 3.0 19.7 16 2.9 36.2 2.5 26.6 2.7 31.4 2 2.8 25.1 3.0 16.1 2.9 20.6 29 2.6 20.6 3.0 23.3 2.8 21.9 28 2.6 25.4 2.6 25.4 26 2.2 17.4 2.0 19.8 2.1 18.6 46 1.5 37.1 1.0 28.2 1.3 32.7 21 3.1 18.2 3.1 18.2 4 1.8 16.7 1.8 16.7 Grand av 4.5 22.3 3.0 18.6 3.7 20.7

21 The number of stems comprising every bunch was measured at both sites and the average stems/bunch reported here. In contrast to the yield differences between the sites, there is more consistency in varietal response in the stem numbers required to make a bunch. Three notable exceptions are varieties 32, 20 and 34, three of the top performing varieties in the Southeast in terms of total weight. These varieties produced dominant terminal flower heads in the Southeast compared to more lateral branches at the Lenswood site. Stems tended to be larger and thicker with high flower numbers per stem, and fewer stems per bush. Fewer flowering stems were therefore needed to produce a bunch.

This example illustrates that there is some variability in flowering stem form in varieties, which is particularly true in the first year of production. Early floral initiation of terminal flowers will lead to a dominant central leader form with a high concentration of flowers near the top of the stem as compared to a weaker leader which allows for more concentration of flowering in laterals.

The combined averages of bunch characteristics are also reported in this table to reduce variability between sites and allow varietal comparisons. Varieties marked with an asterix all produced bunch yields higher than the average (3.7 bunches) and required fewer than 18 stems to make a bunch. These characteristics would be considered optimum in terms of the current practice of producing compact bunches with a distinct diameter and these varieties would be recommended for top yield.

Stem numbers per bunch is a harvest efficiency issue, with varieties requiring over 25 stems bunch needing reassessment or perhaps a change in management practices to produce a different form of stem for more efficient bunching. As longer stem lengths are required by markets, the standard form of the ixodia dried bunch should be reassessed. In line with other flower crops, a 7-10 stem bunch in the 50-60cm stem length range would be more efficient to produce and may present a better product. Varieties cultivated for use as fresh flowers should be marketed in a different form to traditional dried ixodia to emphasise the differences in use and value. This issue will be discussed further in the commercialisation section.

2.4.3 1997/99 Harvest data comparisons – Lenswood

Tables 2.6 and 2.7 summarise the results of analyses run on the top 15 performing varieties at the Lenswood site, comparing the two harvest years. The LSD values are listed in each category for calculation of significance between values. The average yield for all varieties assessed in Year 2 was 1341g or 3.9 bunches which is n.s. different to the average from Year 1 of 1352g or 4.6 bunches. In the analysis of the top fifteen varieties, Year 2 mean yield (1845g) compares similarly to Year 1 (1855g).

22 Table 2.6 Summary of yield results presented as total harvest weight in grams and number of commercial grade bunches for fifteen leading varieties. Results are presented from both harvest years and as combined data.

Total Harvest (grams) Total Bunches Variety Geo Yr 1 Yr 2 Combined % change Yr 1 Yr 2 Combined % change 34 N 2045 3393 2719 65.9 6.8 8.4 7.6 24.1 40 H 2099 3650 2321 73.9 8.9 15.0 9.8 68.2 17 MR 1950 2675 2313 37.2 5.5 7.0 6.3 27.3 3B N 2149 2157 2152 0.4 6.8 5.5 6.2 -18.5 11 N 2398 1441 2016 -39.9 8.7 3.3 6.6 -62.0 32 DC 1995 1780 1888 -10.8 6.1 5.5 5.8 -10.2 24 N 1776 1935 1844 8.9 4.5 5.3 4.9 17.9 48 H 1812 1743 1784 -3.8 7.0 4.5 6.2 -35.7 31 N 1388 2565 1780 84.9 4.8 7.0 5.5 47.4 23 N 1745 1765 1758 1.1 5.0 4.6 4.8 -7.5 36 H 1858 1498 1738 -19.3 5.3 3.8 4.8 -28.6 10 N 1880 1473 1676 -21.7 6.5 5.0 5.8 -23.1 33 DC 1817 1343 1627 -26.1 6.8 4.4 5.8 -35.2 8 N 1435 1485 1460 3.5 5.3 5.5 5.4 4.8 22 N 1375 1532 1442 11.4 5.1 5.0 5.0 -1.2 All Vars 1352 1341 4.6 3.9 LSD 924 1538 819 3.5 4.1 2.6

Table 2.7 Summary of yield results presented as weight in grams in two stem grade classes (Class 1=30cm (Yr1), 40cm (Yr 2) and Class 2= 40cm (Yr1), 50cm (Yr 2)) for fifteen leading varieties. Results are presented from both harvest years and as combined data.

Yield in Stem Class 1 (grams) Yield in Stem Class 2 (grams) Variety Yr 1 Yr 2 Combined % change Yr 1 Yr 2 Combined % change 34 1243 763 1003 -38.6 803 2630 1716 227.7 40 1023 3180 1331 211.0 1077 470 990 -56.3 17 1450 1215 1333 -16.2 500 1460 980 192.0 3B 1006 1010 1008 0.4 1143 1147 1144 0.4 11 1511 725 1197 -52.0 888 716 819 -19.3 32 968 1645 1306 70.0 1028 135 581 -86.9 24 1058 755 928 -28.6 719 1180 916 64.2 48 1117 1148 1129 2.8 695 793 728 14.1 31 1135 545 938 -52.0 253 2020 842 700.0 23 1475 723 973 -51.0 270 1043 785 286.1 36 1191 1163 1182 -2.3 667 503 626 -24.6 10 1125 673 899 -40.2 755 800 778 6.0 33 842 1346 1042 59.9 975 975 -100.0 8 1230 613 921 -50.2 205 873 539 325.6 22 1064 660 891 -38.0 311 872 551 180.1 LSD 901 778 617 635 1182 196

In Total Harvest, varieties are listed in order of their combined yields, with variety 34 significantly higher than all other varieties except 40, 17, 3B and 11. These top yielding varieties showed significant increases in yield in their second year with the exception of variety 11, the top yielding variety in its first year of harvest which showed decreased yield in the second year. A total of 6 varieties in this analysis showed a depressed yield in the second harvest which is an indication that these varieties are sensitive to over-harvesting or cutting of

23 long stems. The results of Yield of Stem Class 2, the long grade stems, show that these same varieties had a reduction in long stems from Yr 1 to Yr 2 of 19 to 100%.

Varieties 34, 17, and 31 had a high percentage of short stems in Yr 1 but subsequently increased in both total yield and yield of high stem lengths in the second year, which is the overall trend of all the varieties assessed.

The results presented in these tables can be used to compare performance of new varieties or varieties currently under cultivation. Comparisons with average yields of all varieties in either the first or second years of harvest, and with high yielding varieties from the same geographic origins can be used to interpret potential of new varieties.

2.4.4 Extended trial – Varieties 60- 67

The results from the extension of the main field trial (Varieties 60-67) over two years are summarised in Table 2.8. The data presented here is fresh weight of harvestable stems as total weight (TH) and in two stem length classes, 40 cm and >50 cm. In year one, TH of varieties 61 and 62 are significantly higher than all other varieties with the exception of varieties 63 and 60. Variety 61 produced the highest weight of stems in the tall class, 50 cm or greater, and this value is significantly higher than all other varieties except 63. The total yield of var 61 in year 1 represented 7 commercial grade bunches (data not summarised here) which is double that produced by var 63 although there is no significant difference in TH weight.

Table 2.8. Comparison of two years data (1998/99 and 1999/00) of stem classes and total yield (TH) for Varieties 60-67 at the Lenswood site. Data is presented as fresh weight in grams.

Year 1 Weight (gr) Year 2 Weight (gr) Increase Variety Geo 40cm 50cm TH 40cm 50cm TH Yr1-Yr2

60 N 342 626 b 968 bc 390 b 620 abc 1009 bc 4.2 % bcd 61 N 560 b 1129 a 1689 a 661 b 30 c 683 c -59.6 % 62 KI 1023 a 518 b 1541 a 688 b 451 bc 1139 bc -26.0 % 63 N 429 bc 715 b 1144 ab 1620 a 840 ab 2477 a 116.5 % 64 E 250 cd 0 250 d 758 b 0 790 bc 216.0 % 65 N 178 d 454 b 631 bcd 66 H 410 bc 225 b 430 cd 410 b 1175 a 1585 b 268.6 % 67 G 403 bc 191 b 594 543 b 740 ab 970 bc 63.3 % bcd

In the second year, yield of var. 63 is significantly higher than all other varieties in TH and stems in the 40 cm length grade. Yield (TH) of variety 66 is significantly higher than variety 61 which has decreased in TH 60 % over year one. It is probable that high yield in the first year, particularly of stems 50cm and greater, suppressed regrowth in this variety and affected yield in the second year.

24 It is useful to look at combined data from both years to assess varietal performance taking into account suppression of yield that may be occurring from high first harvest. All parameters and interactions are significant, the level of significance is summarised in Table 2.9.

Table 2.9. Level of significance in interactions between year and variety in two stem length class and total harvestable weight.

Significance TH <0.0001 Year*Var <0.0001 40 cm Wt <0.0002 Year* Var <0.0004 50 cm Wt <0.0002 Year* Var <0.0004

TH means increase in Year 2, confirming the predicted increase in yield with increase in plant size. The weight of tall stems (> 50 cm) over all decreases in comparison to the 40cm class. Exceptions to this are varieties 66 and 67 which have low first year yields and then have a high percentage of tall stems in their second year. There are strong Year* Variety interactions in all categories. High yielding first year varieties show a decrease in the mean weight in tall stems in their second year which is consistent with predicted stem length depression with heavy harvesting. This highlights the potential of managing stem length to suit market requirements for tall fresh stems or shorter dried bunches in either the first or second year of harvest for a particular variety.

Table 2.10. Mean total fresh weight yield and yield of stems in two length classes of Varieties 60-67 over two years of harvest.

Year Mean Wt 40cm Wt 50 cm Total Har 1 472.45 546.91 1005.47 2 807.92 523.13 1297.75

When analysing combined yield data from both years, variety 63 in Year 2 is significantly higher than all other varieties by year combinations in TH. Variety 61 in Year 1 is significantly higher than most combinations apart from 63 in Years 1&2 and 66 in Year 2 (Table 2.10).

Varieties 63, 61 and 66 were replanted in further trials in 1999, variety 63 and 61 on the basis of first year performance and variety 66 on its outstanding appearance and uniqueness as a double form.

25 2.5 Post harvest – fresh flower evaluation

The results of vase-life assessments for all varieties in either Year 1 (1997-98) or Year 2 (1998-99) are summarised in Table 2.11. The average mean vase-life of the 74 varieties assessed over two years was 11.3 days with means per variety ranging from a low of 5.2 to a high of 23.3 days. Thirty-three varieties had a mean vase-life of 12 days or better and of these, 50% were from late maturing varieties which are harvested in the period Jan-Feb.

Variety is highly significant in the analysis and there are some significant differences between year results in the same variety. Varieties such as 10, a high yielding form, show n.s. differences in vase-life performance in the two years which is significantly below the average of all varieties. Such performance would indicate that this variety would only be suitable for dry flower production. Var. 11, a similar Nelson form, also shows n.s. differences between years but is also n.s. different from the average, adequate vase-life for fresh flower use.

Because variety differences are significant, all data is presented here, so that vase-life performance of particular varieties can be referred to in discussion of overall assessment. To summarise vase-life performance, varieties are once again grouped by geographic origin and presented in Table 2.12.

26 Table 2.11 Summary of vase-life (days) of fresh stems of 74 selected varieties of Ixodia achillaeoides harvested Nov to Feb (months 11-2) over two years, 1997-8 (Year 1) and 1998- 9 (Year 2).

Variety Geo Month Year Vase-life Stderr Days 1 2 H 12 1 11.33 0.9428 2 3 N 12 1 7.83 0.1667 3 5 H 1 1 14.27 0.3446 4 5 H 2 2 10.00 0.0000 5 6 H 2 1 13.08 0.4345 6 7 H 1 1 13.67 0.5404 7 7 H 1 2 12.83 0.3860 8 8 N 12 1 11.44 0.3768 9 10 N 11 1 7.56 0.5191 10 10 N 11 2 7.00 0.0000 11 11 N 11 2 11.00 0.2582 12 11 N 12 1 12.44 0.3979 13 14 G 11 1 9.50 0.5000 14 14 N 11 1 10.00 0.5774 15 14 G 12 1 10.00 0.5528 16 15 G 12 1 14.50 0.9003 17 16 N 1 1 12.78 0.4888 18 17 MR 12 1 14.00 0.5774 19 17 MR 12 2 15.50 1.0247 20 18 G 2 1 7.53 0.1333 21 19 G 2 1 6.67 0.1260 22 20 G 1 1 13.67 0.8819 23 21 N 11 2 9.17 0.4773 24 22 N 11 2 12.00 0.6853 25 22 N 12 1 10.42 0.7226 26 23 N 11 2 10.00 0.0000 27 23 N 12 1 11.89 0.5122 28 24 N 11 2 15.67 0.9888 29 24 N 12 1 12.75 0.9385 30 25 N 11 1 10.06 0.4535 31 26 N 11 1 12.56 1.3449 32 28 G 1 2 9.00 0.0000 33 28 G 12 1 13.43 0.9241 34 29 N 12 1 10.17 0.2973 35 30 N 12 1 9.58 0.3580 36 31 N 12 1 11.33 1.1156 37 32 DC 12 1 15.89 0.9639 38 33 DC 1 1 16.53 0.7860 39 33 DC 1 2 12.00 0.0000 40 34 N 11 2 8.33 0.4410 41 34 N 12 1 5.17 0.4014 42 35 H 2 1 10.20 0.2619 43 36 H 2 1 7.56 0.1205 44 37 H 2 1 10.42 0.1487 45 38 H 2 1 8.27 0.5474 46 39 H 2 1 8.58 0.1930 47 40 H 1 1 12.44 0.5253 48 40 H 1 2 12.00 0.0000

27

Variety Geo Month Year Vase-life Stderr Days 49 41 H1 1 10.39 0.2306 50 42 H1 1 13.61 0.3146 51 43 N111 14.00 0.0000 52 44 KI 1 1 11.44 0.3768 53 45 KI 1 1 7.00 0.3893 54 46 KI 1 1 11.07 0.4505 55 47 DC 12 1 17.00 0.7140 56 48 H2 1 9.72 0.2658 57 48 H2 2 13.00 0.0000 58 49 G2 1 10.92 0.9167 59 50 G2 1 10.00 0.3086 60 60 N112 9.00 0.0000 61 61 N111 14.08 0.0833 62 62 KI 2 1 10.89 0.2003 63 63 N112 8.36 0.3377 64 64 E102 11.33 0.6667 65 67 G2 1 10.89 0.5386 66 72 G2 1 9.44 0.2422 67 76 G2 1 6.50 0.3416 68 110 H2 2 9.33 0.3333 69 112 H2 1 9.78 0.4006 70 112 H2 2 12.67 1.1738 71 114 H2 2 8.00 0.0000 72 115 H2 2 9.83 0.1667 73 142 DC 1 2 23.33 0.9189 74 160 DC 1 2 16.44 0.7837 75 161 DC 2 2 16.67 0.2357 76 223 N112 8.50 0.2236 77 224 N112 11.00 0.2582 78 225 N112 10.75 0.1306 79 231 N112 8.00 0.0000 80 232 N121 19.67 1.3162 81 280 N112 10.67 0.3333 82 280 N122 10.33 0.6667 83 313 G2 1 9.17 0.3073 84 316 G2 1 9.33 0.3333 85 331 G2 1 14.00 0.7882 86 352 G1 2 11.67 0.3333 87 411 A2 2 9.44 0.1757 88 413 A2 2 12.00 0.7071 89 415 A2 2 12.56 0.1757 90 416 A2 2 9.56 0.2422 91 518 G2 1 16.17 0.4014

28 Table 2.12 Summary of mean vase-life (days) of all varieties grouped by Geographic origin.

Geo. class Observations Vase-life (days) Deep Creek 72 16.8 a Mount Richmond 9 15.0 a Eyre 9 11.3 b Adelaide Hills 258 11.1 b Angelsey 36 10.9 b Nelson 311 10.8 b Grampians 173 10.7 b Kangaroo Island 44 10.2 b

Varieties from the Southern Adelaide Hills region designated Deep Creek have significantly higher mean vase-life than the other major classes designated Adelaide Hills and Nelson where the majority of current cut-flower varieties originate. Varieties 33 and 32 with higher than 15 days vase-life have also been ranked outstanding in terms of yield and floral characteristics.

The large flowered, coastal Mt Richmond forms also show high potential for use as fresh flowers, although most forms evaluated in our program where dwarf and did not produce the tall straight stems that are desirable for a quality fresh product. The exception to this is variety 17 which performed well over 2 years (mean 14 and 15.5 days) and is a high-yielding long- stemmed variety.

Using vase-life as a prime indicator of success of fresh product performance, our results show that the Nelson forms (which are currently most frequently used for fresh flower exports) are most variable in performance of the main geographic groups and that varietal screening for vase-life is perhaps more critical than in other groups. This variability may be due to the rapid stem growth elongation occurring in spring (see Figure 2.4) which results in a softer shoot than the slower maturing varieties harvested in late summer. Presumably a softer shoot, harvested on a warm day is more vulnerable to water loss and deterioration than woodier material that has developed in the more predictable weather patterns experienced in summer production. Spring weather in South Australia and Victoria is characteristically highly variable with strong changes in mean temperatures which impact on flower quality. Coastal cool-climate growing conditions would be strongly recommended for production of early Nelson varieties which are being produced for fresh flower markets.

Nelson varieties which performed well in vase-life trials include: 24, 11, 61 and 8. Of these, varieties 24 and 11 are the most consistent high-yielding varieties assessed. Nelson forms have other quality characteristics which make them desirable as fresh cut flowers. They produce the highest number of long stems ( >60 cm in their first year of production), there are many distinct spray forms with reduced stem numbers/bunch and they produce less of the sticky resins that can rub off on workers and florists hands.

29 Adelaide Hills forms characteristically produce straight tall stems with large terminal flower heads with very showy tightly packed flowers. Stems and leaves are an intense dark green which contrasts well with the white petals and either light yellow or maroon flower centres. High-yielding varieties assessed with higher than average vase-life and excellent potential for fresh flower production include: 40, 48 and 36.

2.6 Stem growth measurements

The results of the fortnightly measurements of stem growth over 18 months are summarised in Figures 2.4. and 2.5. In the first figure, variety 25, a Nelson form, is harvested in mid- November and commences new growth in early Feb. in contrast to the Hills form, var. 41 which is harvested in late January and commences new growth in mid-March. The Nelson form has a higher rate of growth through the autumn/winter period than the Hills form, with a distinct growth flush in late September prior to flowering. The Hills variety has a longer growth flush that peaks in early mid-November and stops during summer months until harvest. Variety 32, a hybrid form, has intermediate characteristics.

Figure 2.5 compares the earliest harvested var. 43 with a late harvested Grampians form. Both varieties show more sustained growth over the year than the varieties in the previous figure, however, var. 43 has a period of rapid stem elongation in early September, similar to the Nelson variety. Soft growth at this time of year can make these varieties vulnerable to frost and unseasonable weather changes in interior climates. They are, however, protected from prolonged exposure to heat injury which may be experienced by new growth in the later months.

An understanding of stem growth patterns for a variety can assist in management practices to optimise yield and quality. For example, fertiliser applications in spring are more critical for Hills and Grampians varieties which experience the majority of their stem growth elongation in a short period prior to flower initiation. Fertiliser applications after harvest are particularly critical for SA and Nelson varieties which establish considerable stem length over late summer/autumn periods.

30

12.0

10.0

Year 1 Year 2 8.0

6.0 25 41 32 4.0

2.0

0.0

8 9 97 9 98 98 98 997 998 998 998 998 -19 1 -1997 -1 -19 -1 -19 -19 -199 p t- v c-1997 r-1 r-19 n l g p v c e p p u ec-1 e eb-1998 A u e -F - -Au 2-S 6-No 3 4-Mar 2 3-J 7-J 4 0-S 2-No 30-Sep-199729-O 2 30-D 29-A 1 15-Oct-19981 15-D

Figure 2.4 Fortnight growth increases in stem length (cm) plotted over an 18 month period from 3 varieties grown at the Lenswood site.

12.0

10.0

Year 1 Year 2 8.0

43 6.0

49 4.0

2.0

0.0

7 7 8 8 97 98 98 98 9 998 9 998 998 99 999 199 1997 1999 - -1 r-1 -1 -1 -19 t-199 -1 - ct- b a g-1 c v ep-1997 ov-1 e pr Jul u ep-19 o ec-1998an -S -O N F M A A N D J 2-Sep-199 3- 4- 2- 3-Jun-19987- 2- 30 29 26- 30-Dec 29-Apr 4- 10-S 15-O 1 15- 21-

Figure 2.5. Fortnightly growth increases in stem length (mm) plotted over an 18 month period from 2 varieties grown at the Lenswood site.

31 2.7 Variety Descriptions and Identification

2.7.1 Harvest date records

Flowering dates for all varieties over 3-4 years of harvest in our trials is summarised in Appendix. Variation of 7-10 days for peak harvest date is common over different years, but the order of bloom of individual varieties remains similar each year. Flowering date sequence is useful tool in identifying varieties and geographic origins of a variety if it is unknown. Placing an unknown variety into a geographic grouping is helpful in understanding many other characteristics of a variety and its performance under cultivation. Figure 2.4 summarises the period of flowering for all assessed varieties (averaged over four years) grouped by their geographic origin. In the case of forms designated SE (early Southeast commercial variety) and MR (Mt. Richmond), only one variety is represented.

G

A

H

E

KI Variety DC

MR

N

SE 8-Jan 7-Feb 9-Dec 9-Nov 20-Oct 30-Oct 18-Jan 28-Jan 17-Feb 27-Feb 19-Dec 29-Dec 19-Nov 29-Nov

Date average

Figure 2.6 Average flowering periods over four years of all varieties at Lenswood site grouped by geographic origin. The Bar represents the grand mean of all varieties in each group.

The Nelson (N) varieties are distinguished from all other varieties by their early flowering time. It is understandable that these varieties have been extensively used in the past few years by the wildflower industry as early market periods are generally favourable to supply domestic markets and spring harvest corresponds more closely to other wildflower crops which may be produced in a mixed wildflower enterprise.

32 At this site in the Adelaide Hills there is a distinct gap in production in an important fresh market period in early to mid December. For this reason the varieties designated DC are particularly important for potential fresh Christmas sales. The Deep Creek forms are distinct geographically isolated population in the lower Adelaide Hills region with intermediate characteristics between Hills (H) and some Kangaroo Island populations. Two hybrid forms are also grouped in this class

2.7.2 Floral descriptions

The results of floral measurements of overall size, size of centres and # petals is presented by individual variety, grouped by geographic origin in Figures 2.7 to 2.10.

In comparing the first two tables of Nelson and Adelaide Hills varieties, the overall size of the Nelson flowers is consistently greater that the Adelaide Hills forms, although there is more variation in the size of the flower centres. As small centres are often rated as desirable in appearance criteria, then varieties with a high ratio of flower size to centre size such as 8, 9 and 22 would be more desirable than vars. 11 or 224. Petal count is highly variable in the Hills forms which are represented by some varieties which are classified as ‘doubles’, although the petal count here includes only primary petals and not the petaloids which comprise the centres in many double flowers.

Flowers from the Grampians region (Figure 2.9) are composed of the small flowering forms from the southern regions (vars. 18-20) and several isolated populations throughout the range which vary in size up to the double var. 76, which is as large as a Nelson form. Two large flowering forms (vars. 43 and 17) are presented in Figure 2.10. which groups representatives of other geographic collections. The Angelsey forms have a desirable flower to centre size ratio and the Eyre form (var. 64) illustrates the typical small, low petal count flowers that are typical of coast forms from the Eyre and Yorke peninsulas.

Nelson Region

16 25 14 20 12

10 15 8 6 10 Diameter (mm)

4 Number of Petals 5 2 0 0 2 8 9 23 11 10 22 30 60 34 26 24 29 63 31 25 225 224 230 63x22 Variety Flower diameter Centre diameter # petals

Figure 2.7 Flower descriptive data for twenty selected varieties and hybrids originating from the Nelson region

33

Adelaide Hills Region

16 25 14 20 12

10 15 8 6 10 Diameter (mm)

4 Number of Petals 5 2 0 0 6 5 7 66 36 48 38 39 37 41 40 35 42 33 32 47 115 110 114 160 Hills Deep Creek Variety Flower diameter Centre diameter # petals

Figure 2.8 Flower descriptive data for twenty varieties originating from the Adelaide Hills region including 4 varieties and hybrids designated Deep Creek, from the lower Fleurieu Peninsula.

Grampians Region

14 35 12 30 10 25 8 20 6 15 4 10 Diameter (mm) 2 5 Number of Petals 0 0 18 20 19 16 50 67 14 28 28 49 15 76 352 354 Variety

Flower diameter Centre diameter # petals

Figure 2.9 Flower descriptive data for fourteen varieties originating from the Grampians region.

34 Other Locations

20 30 25 15 20 10 15 10 5

Diameter (mm) 5 Number of Petals 0 0 4 43 64 62 44 45 46 17 413 423 427 518

South Angelsea Eyre Penninsula Kangaroo Island Mt Richmond Unknown East Variety (number and region) Flower diameter Centre diameter # petals

Figure 2.10 Flower descriptive data for ten varieties originating from four geographic areas designated, Anglesea, Eyre, Kangaroo Island and Mt Richmond and two varieties of unknown origin.

Comparison of Geographic Forms

18 HillsDeep Creek Nelson 25 16 14 20 s ) 12 15 10 8 10 6 Diameter (mm Number of Petal 4 5 2 0 0 40 48 66 AV 33 160 47 AV 11 24 63 AV Variety Flower diameter Centre diameter # petals

Figure 2.11 Average and representative flower descriptive data for varieties originating from the three primary geographic groups in our study. Standard errors of averages are indicated by vertical bars.

Figure 2.11 presents the average data from all varieties assessed in the Adelaide Hills, Deep Creek and Nelson geographic origin classes. Representative varieties from each of these

35 classes is also presented for comparison. Standard errors of the averages are represented by vertical bars. The Adelaide Hills forms have an average flower diameter of 9 mm and centre of 2.6 mm. By comparison, the Nelson varieties average 12 mm in diameter with a centre of 3.5 mm. Deep Creek varieties are intermediate in form with an average of 10.3 mm and centre of 2.9 mm but they are more variable as a group than the other groups as is indicated by the error bar. The data presented here indicates how floral data can be used to identify or distinguish varieties and geographic forms and particularly in describing new varieties and hybrids which may be required in seeking Plant Breeder’s Rights protection.

2.7.3 Colour data from all the assessed forms

Histogram of Flower Centre Colours

25

20

15

10 number

5

0 155b 155c 158a 161a 161b 161c 162a 162b 162c 163b 163c 199a 199c 199d RHS colour

Figure 2.12 Summary of mean flower centre colours of dried samples of all assessed varieties using RHS colour charts, 1995.

The results of flower centre records presented in Figure 2.12. show that colours range from a white (155a) through to brown (199a) with the majority in the 161-162 beige to ochre range. The largest variation in centre colour comes from the Grampians group, with Nelson varieties being the most similar with the 161a colour group predominating. There appears to be a preference for light coloured centres or extremely dark ones among growers and florists. Fresh flower colours predominate in the yellow groups (8-11) or greyed purple group (183- 187).

PBR Comparative Trials

Measurements were made at harvest of a range of vegetative and floral characteristics of three varieties of Adelaide Hills origin and one of hybrid origin (33). One commercially known variety (5) was used as a comparator for two selections from our development program (40, 41).

Measurements made included: total # of flower in the inflorescences, # of flowers in the top 5 cm of the inflorescence, flower and flower centre diameter, stem diameter at 3cm, angle of laterals from the central axis at 10cm and 20 cm, fresh weight of stem (40cm) and number and length of laterals at 20cm from the top. The results are shown in Tables 2.13 and 2.14.

Table 2.13 Comparison of stem characteristics of flowering shoots of 4 varieties of Ixodia achillaeoides.

36

Mean Fresh Stem Latera Flower Centre Length at Variety weights (gr) diameter l # diameter diameter 20 cm (mm) (mm) 5 55.00 a 7.47a 6.4 9.0 a 2.75 a 4.82 a 40 54.19 a 5.52 c 5.7 9.3 a 2.67 a 22.15 b 33 41.25b 4.67 d 6.6 7.7 b 2.08 b 17.45 b 41 41.22 b 6.57 b 5.9 9.0 a 2.42 ab 5.86 a

w Mean separation within columns by LSD, P< 0.05, no letters indicates n.s.

Table 2.14 Comparison of mean terminal and total flower numbers and angle of branching of laterals from main axis at 10 and 20 cm from the top of flowering shoots of 4 varieties of ixodia.

Mean Flower # Total Angle of branching from main Variety Top 5 cm Flower # axis w At 10cm At 20 cm 5 153.3 b 292.1 bc 44.95 a 39.80 ab 40 327.1 a 416.9 a 37.95 ab 46.55 a 33 94.1 c 356.0 ab 34.45 b 34.45 b 41 170.9 b 261.7 cd 41.50 ab 42.20 ab

w Mean separation within columns by LSD, P< 0.05

By comparing stem characteristics, varieties 33 and 41 are shown to be significantly different than the comparator var. 5 in weight and stem diameter and var. 40 different to the comparator in stem diameter and length of laterals at 20 cm. When comparing flower characteristics, var. 40 proves to be a useful comparitor with significant differences to all varieties in terminal flower numbers and all but one variety in total flowers. Angle of lateral branching is a less useful parameter to separate these varieties but at least one variety is significantly different from all others at each position.

2.8 Assessment of dwarf plants for pot plant production

A range of trials were conducted from 1997 to 2000 to assess dwarf ixodia varieties as flowering and foliage pot plants. As this was not in the original brief of this research program, original collections did not concentrate on enlarging or improving the few dwarf lines that we possessed. Fortunately, the dwarfs we had continue to show high potential for pot plant development and with the wide range of trials in which they were incorporated, now give us a good understanding of how each variety can be manipulated for flowering pot plants. New assessions, hybrids and seedlings have subsequently been added to our collection and traditional cut flower varieties have been treated with growth retardants for assessment. Following are three trials that illustrate the manipulation of flowering and size that can be achieved in the development of ixodia pot plants.

37 2.8.1 Length of coolstore treatment affects growth and flowering

In this trial, four dwarf varieties and one standard cut flower variety (62) were grown for 16 weeks from cuttings and potted into 155 mm squat pots prior to treatment. Varieties 534 and 615 had been previously assessed as dwarf plants, the others were new assessions with potential to be grown as flowering pot plants. Height and size measurements were recorded at the start of the trial on 31.12.98 and plants were given the following treatments: A) 4 weeks in 14°C coolstore, SD lighting B) 8 weeks in 14°C coolstore, SD lighting and C) 8 weeks in 22- 26°C greenhouse (LD conditions). There were 8 reps per treatment and a total of 120 plants. After treatment plants were all grown on for a period of 8 additional weeks in a controlled environment greenhouse (20-26°C) on ebb and flow beds under natural light until flowering. Growth and flowering data were recorded on 27.4.99. Data was analysed as a treatment x variety factorial by general AOV using Statistix v.3.1.

The results in terms of growth are presented in Table 2.15. In all cases, CS treatment for 8 weeks significantly reduced height of plants over GH controls, and 4 weeks of treatment significantly reduced height only in varieties 62 and 428. Significant size reductions with 8 weeks of CS treatment were seen in Vars. 62, 428 and 534 and with 4 weeks of CS in Vars. 62 and 428.

Table 2.15 Results of coolstore treatment on growth of 5 ixodia varieties grown as flowering pot plants.

Heightw Height x Widthz Variety Variety Treatment 62 427 428 534 615 62 427 428 534 615 A 4 weeks in 4205. 1873. 1762. 2214. 2082. 78.3 46.7 39.0 45.3 47.2 Cool Store 5 2 7 0 3 B 8 weeks in 3651. 1792. 1294. 1780. 1772. 76.8 45.8 35.0 40.0 41.3 Cool Store 3 0 5 3 8 C 8 weeks in 5209. 1946. 2120. 2154. 1995. 85.2 41.2 44.0 44.2 47.7 20-24°C GH 0 5 2 3 8 wLSD (0.05) for mean separation between values = 3.8021 zLSD (0.05) for mean separation between values = 323.9988

Comparing varieties across treatments, Var. 428 is significantly shorter in height and smaller in size than all other varieties with the 8 week coolstore treatment. Variety 62 (a standard cut flower variety) is significantly taller than all other varieties across all treatments.

38 Table 2.16 Results of coolstore treatment on flowering of terminal and lateral branches of 5 varieties of ixodia grown as flowering pot plants.

Flowering of terminalsw Number of flowers on lateral branchesz Variety Variety Treatment 62 427 428 534 615 62 427 428 534 615 A 4 weeks in 0 0 3 14 10 0 0 1.2 0 0 Cool Store B 8 weeks in 6.2 3.2 3.2 16.2 10.8 7.8 1.8 9.5 0 13.8 Cool Store C 8 weeks in 0 0 0 0 0 0 0 0 0 0 20-24°C GH wLSD (0.05) for mean separation between values = 2.0059 zLSD (0.05) for mean separation between values = 3.5950

Table 2.16 presents flowering data for these plants, in two categories: flowering terminals and flowers on lateral branches. Flowering of lateral branches has been shown to be enhanced by SD treatment in comparison to LD treatment which will reduce time to flower initiation produce flowers only on the terminals (Weiss et al, 1996). In these results, all varieties show a significant increase in flowering of terminal branches with 8 weeks of coolstore treatment over controls. In varieties 428, 532 and 615, 4 weeks of coolstorage also produced significant flowering over the controls. Lateral branch flowering is significantly enhanced by 8 weeks of CS treatment compared to both the control (see Figure 2.13) and 4 week treatment in all varieties except var. 428 where there is n.s. difference between the 4 and 8 week coolstore treatment.

Figure 2.13 Results of SD treatment at 14°C on Var 534. Plant on left in the Control, then 4 week and 8 week treatments.

These results confirm the results of Weiss et al., that SD treatment enhances lateral branch flowering, which is a desirable characteristic for a well rounded flowering pot plant. As well these results show that there are varietal differences in response time to flower initiation as varieties 534 and 615 had significant number of flowering terminals over the other varieties with only 4 weeks of coolstore treatment which was not significantly increased by the longer 8 week treatment. With variety 615, however, a large number of lateral branches also flowered with the 8 week treatment which would be recommended in preference to the 4

39 week for enhanced flowering. Figures 2.14 and 2.15 illustrate flowering pot plant products resulting from this trial.

2.8.2 Coolstore treatment enhances compactness

In this trial, four dwarf varieties and one standard cut flower variety (63) were grown for 10 weeks from cuttings and potted into 125 mm squat pots prior to treatment. There were 8 reps per treatment. Height and size measurements were recorded at this time and plants were given treatments of : A) Bonzi® growth retardant (20ml/l solution, 50ml drench per pot) plants grown in 20-24C greenhouse, B) Bonzi® drench treatment as above, plants grown in 14°C coolstore under SD conditions for 8 weeks and C) No drench, plants grown in coolstore, 14°C, SD, as above. Again, the data was analysed as a treatment x variety factorial by general AOV using Statistix v.3.1

The results of growth measurements after 8 weeks of treatment are presented in Table 2.17. In all varieties, there is n.s. difference in height or size between treatments B and C, Bonzi® treated and untreated plants grown in the coolstore. There is a trend towards dwarfness in varieties 615, 530 and 63, which suggests that a higher rate of Bonzi® may be needed to significantly reduce size. In all varieties there is a significant reduction in height and size with the CS treatments (B and C) compared to the Greenhouse grown treatment, A. The main agent of dwarfing and size reduction in this trial was the coolstore treatment which is used to induce flowering.

Table 2.17 Results of Bonzi® and coolstore treatments on growth (change in height and size (height x width) in cm) of 5 varieties of ixodia grown as flowering pot plants.

Change in Heightw Change in Size (Height x Width)z Treatment Variety Variety 615 530 534 63 615 530 534 63 A Bonzi®, 716. GH 8.3 7.0 8.8 18.7 364.8 372.5 325.8 5 20-24C B Bonzi®, CS 201. 2.1 .8 2.4 6.5 70.6 76.1 92.2 14C, 8 weeks 0 C No Bonzi® 239. 2.3 1.1 2.5 6.6 96.2 110.7 87.0 14C, 8 weeks 0 wLSD (0.05) for mean separation between values = 1.1958, zLSD (0.05) for mean separation between values = 58.4587

Comparing varieties across treatments, var. 530 is significantly shorter in height than all other varieties in all treatments but n.s. smaller in size. Variety 63, the cut flower form used as a control, is significantly taller and larger than all other varieties across all treatments. It is interesting to note the large effect that the CS treatment had on the size of a non-genetic dwarf variety. There may be potential to produce a suitable pot plant out of standard cut flower form with appropriate rates of growth retardants and cold treatments.

40 (B) (A)

Figure 2.14 Example of pot plants showing dwarf variety (530) compared to standard cut flower variety (63) and finished pot plant of variety 615.

2.8.3 Bonzi® treatments affect dwarfing

In this trial, three new dwarf varieties and two standard cut flower varieties (62 and 280) were grown for 12 weeks from cuttings and potted into 125 mm squat pots prior to treatment. Height and size measurements were recorded at this time and plants were given treatments of : A) plants grown in 14°C coolstore under SD conditions for 8 weeks B) Bonzi® growth retardant (20ml/l solution, 50ml drench per pot) plants grown in 14C coolstore under SD conditions for 8 weeks C) Bonzi® drench treatment as above, plants grown in greenhouse 22- 26C C) No drench, plants grown in greenhouse 22-26°C.

The results of growth measurements after 8 weeks of treatment are presented in Table 2.18. In all cases, coolstore treatment alone significantly reduced growth in terms of height and size compared to greenhouse grown controls, with the exception of height in var. 62. When a Bonzi® drench treatment was incorporated with coolstore treatment, plants were reduced in size and height but not significantly compared to coolstore treatment alone. Compared to greenhouse grown controls, all varieties were significantly reduced in height and size.

The results of treatment C show that in 3 varieties, 280, 621 and 624, Bonzi® drenches significantly reduce height and size in greenhouse grown plants compared to controls, however, these size reductions may not be large enough to produce a well proportioned pot plant. This treatment was included here because potential exists for producing not flowering foliage plants with some varieties of ixodia that produce a natural topiary shape and have scented foliage.

41 Table 2.18 Results of coolstore and Bonzi® drench treatments on growth (change in height and size (height x width) in cm) of 5 varieties of ixodia grown as flowering pot plants.

Change in Height (cm)w Change in Height x Width (cm2)z Variety Variety Treatment 62 280 621 622 624 62 280 621 622 624 A 14°C, 8 wks 13. 10.4 3.3 4.0 8.4 332.5 412.6 106.6 126.5 90.2 Coolstore 2 B 14°C, 8 wks 11. Coolstore 12.4 2.3 2.6 6.5 223.3 438.9 83.5 123.8 68.9 6 Bonzi® drench C 20-24°C GH 13. 26.2 6.4 5.6 12.9 389.6 1189 157.7 193.3 202.5 Bonzi® drench 7 D 20-24°C 17. 33.2 10.8 8.4 17.5 481.6 1370 314.3 331.9 346.8 GH 2 wLSD (0.05) for mean separation between values = 4.14761, zLSD (0.05) for mean separation between values = 136.9862

2.8.4 Discussion The examples of pot plant trials summarised here illustrate that flowering and size control can be manipulated in ixodia with cold treatments and day length control to enable the production of suitable flowering pot plants utilising dwarf varieties. Ixodia has been shown to respond to Bonzi® growth retardants, however, the effect is varietal dependant and also cumulative with cold treatments in size reduction. Bonzi® rates can also be manipulated to affect size of dwarf and standard cut flower lines of ixodia (data not presented here). A combination of suitable treatments can be developed for each specific variety to enhance flowering and size control for production of flowering and foliage pot plant lines.

The development of these specific protocols for flowering pot plant production is tied to commercialisation of individual varieties and should be sold with the rights for production. Dwarf ixodia varieties have excellent potential for European and North American markets because they can be produced in a cool greenhouse under SD conditions in winter. By responding to daylength and growth retardant manipulation, they can be grown for target market periods or as a rotational crop to utilise cool growing conditions. Some varieties can be grown without the use of growth retarding chemicals, which has added benefits to producers and some target markets that are valuing chemical-free plants. The dark green dense foliage mimics other non-flowering pot plant lines currently popular in these markets and the star-like fine flowers of ixodia further enhance the foliage.

In assessing an individual variety’s potential as a flowering pot plant, other factors such as leaf size and density, foliage colour, flower (capitulum) size and appearance are also important in the overall appearance of the product. The flower size of ixodia blooms are small relative to other members of the Asteraceae used for pot plant production (e.g. Helichrysum sp.), therefore the appearance of individual flowers needs to be optimal (bright white colour, high petal count, relationship of centre to petals) and foliage dense, dark green and preferably perfumed.

42 2.9 Landscape variety assessment

The results of the first year assessment of landscape varieties is presented in Table 2.19. Varieties are sorted from the shortest to the tallest with rankings given in terms of dwarfness or overall size of the plant.

Table 2.19 Results of evaluation of 9 dwarf varieties with potential for landscape use.

Variety Geo Av H Av HxW Dwarfness Size Ranking Ranking 531 KI 35 a 2349 a 1 2 426 AV 36 a 2453 a 2 3 64 E 44 ab 1930 a 3 1 534 KI 50 b 4519 b 4 4 721 MR 56 bc 5367 b 5 6 428 AV 59 c 5069 b 6 5 427 AV 75 d 6857 c 7 8 530 KI 79 de 8319 d 8 9 49 G 89 e 5854 bc 9 7

Three varieties; 531, 426 and 64 are significantly smaller in height and overall size to all other varieties with the exception of varieties 64 and 534. The remainder of the varieties group into an intermediate and larger class, all of which form dense mounding shrubs with the exception of variety 49 which is tall and upright in habit. Results after 24 months of growth will rank floriferousness and longevity and recommendations can be drawn on landscape potential.

43

(A) (B)

(C) (D)

(E) (F)

Figure 2.15 Ixodia variety field assesssment program: Planting new trial in the Southeast First year harvest of an ixodia bush showing size of a bunch Harvest of stock plants grown in comparative trial, 1999-2000. Field portion of landscape variety evaluation Drying shed where bunches were assessed. Weighing fresh bunches and recording data after harvest.

44

(A) (B)

(C) (D)

(E) (F)

Figure 2.16 Pot plant assessment on ebb and flow beds Portion of stock plant collection held in greenhouses Set up of postharvest evaluations Plants undergoing SD low temperature treatment to initiate flowering hand pollination of ixodia stock plants Grower meeting to assess new ixodia varieties at Millicent library.

45 3. Diseases of Ixodia: identification, varietal assessment and evaluation of control methods

3.1 Introduction

Diseases account for significant yield and plant losses in commercial plantings of Ixodia daisy. Over a period of eight years prior to the commencement of this program, numerous pathogen specimens were collected from commercial plantings and identified in the SA Department of Agriculture’s and later SARDI’s Horticultural Plant Pathology Unit. Systemic diseases such as fusarium and verticillium accounted for the most serious losses in plantings, however, no study had been made on the incidence of spread of these diseases or relative susceptibility of the more commonly used varieties. Nematodes were known to be involved with losses of Ixodia in plantings in the South-east of SA and a suspected association with verticillium was thought to account for the greatest losses.

At the start of our program in 1996, surveys of natural populations and commercial plantings were conducted to collect diseases associated with Ixodia and pathogenicity tests were were used to confirmed 7 fungi and 1 nematode as pathogens of Ixodia. 61 cultivars of Ixodia, which were assemnbled as part of the variety development program, were used for screening in inoculation trials, to establish if varieties existed with a relative resistance to the major diseases. The results were utilised in the varietal development program.

Control trials were carried out on two diseases of primary concern to producers. Powdery mildew trials assessed ten treatments including fungicides and biological controls. Control of two phytophthora species were evaluated in greenhouse and field trials over the course of three years. Chemical controls and a management program was developed to these diseases.

3.2 Pathogen identification

Wilting, yellowing and dying plants of Ixodia achillaeoides have been observed in natural and commercial plantings since 1988. Plants exhibiting these symptoms, either collected from sites in the Adelaide Hills and South East of South Australia during 1995 or submitted to the Horticultural Diagnostic service for disease identification, were tested for the presence of disease causing organisms.

3.2.1 Materials and methods

The plants were surface sterilised in 0.4% Sodium Hypochlorite for 3 minutes and rinsed in demineralised water. Pieces from the edge of the diseased tissue were then plated onto artificial media, either Cornmeal agar (CMA), Potato dextrose agar (PDA) Water agar (WA) or Phytophthora specific media (PVP10+). After 5-14 days incubation at 22oC plates were examined and any fungi present was identified by morphological characteristics using standard keys.

Infected leaf pieces were also placed into paper lined trays with wet towelling and enclosed in a plastic bag to create 100% humidity. After 3-7 days incubation at 22oC, pieces were

46 examined and any fungi present was identified by morphological characteristics using standard keys.

Soil was baited for Phytophthora by placing 60-100g of soil in a small tub, filling with demineralised water, and either placing lupin sprouts with the root in the top of the water, or Camellia leaf discs floated on the water. The Camellia leaf discs were removed after 48 hours and plated onto PVP10+. The lupins were examined at 7-10 day for the presence of sporangia in the root, and infected roots were then plated onto PVP10+.

Any pathogenic fungi were subcultured and forwarded to Dr M Priest, mycologist with NSW Agriculture for confirmation of identification, and new records were entered in to the Australian herbarium collection.

Powdery mildew was first observed on a plantation in the Adelaide Hills area of South Australia. The leaves were covered with mycelial and conidial growth with some localised necrosis. The fungus, designated an Oidium sp., appeared to be a new species and was identified by I. Pascoe (Agriculture Victoria, Knoxfield).

The crown gall and phytoplasma were identified by visual inspection and forwarded to Dr K Ophel-Keller, scientist with SARDI or Dr N Habili, AGPROBE Diagnostics, Adelaide, respectively for confirmation.

Any samples with bacteria present were forwarded to the Crop Health Services laboratory, Agriculture Victoria, for identification.

Nematodes were extracted from a 200g sample of soil incubated on Whitehead and Hemming trays for five days at 22°C. Roots were cut into 2cm lengths and transferred to plastic bags for extraction of nematodes by incubation in 3% hydrogen peroxide at 22°C for five days. Nematodes were identified by Dr G Walker, nematologist with SARDI, and entered into the Waite collection after confirmation of identification by Dr F Reay, nematologist with the University of Adelaide.

3.2.2 Results

12 Fungi, 12 nematodes, 2 bacteria and a phytoplasma were all recovered from the Ixodia tested and are summarised below in Table 3.1.

47 Table 3.1. Symptoms of disease and the associated fungi identified on Ixodia achillaeoides.

Symptoms Fungi recovered Identified by and collection number* Leaf lesions and Alternaria alternata B Hall damage Stemphyllium sp. B Hall Oidium sp. (Powdery mildew) I Pascoe (VPRI20670) Capnodium sp. (Sooty mould) B Hall Tip dieback Botrytis cinerea B Hall Fusarium oxysporum M Priest (DAR72349) Verticillium dahliae M Priest (DAR69284) Stem staining Fusarium oxysporum M Priest (DAR72349) Fusarium tabacinum M Priest Fusarium acuminatum M Priest (DAR72352) Verticillium dahliae M Priest (DAR69284) Stem lesions Botrytis cinerea B Hall Alternaria alternata B Hall Root rotting Fusarium oxysporum M Priest (DAR72349) Phytophthora cryptogea M Priest (DAR72351) Phytophthora erythroseptica M Priest (DAR72350) Rhizoctonia solani B Hall Pythium sp. B Hall Stem cankers Phytophthora cryptogea M Priest (DAR72351) Phytophthora erythroseptica M Priest (DAR72350) Rhizoctonia solani B Hall Soil from root Phytophthora cryptogea M Priest (DAR72351) zone declining Phytophthora erythroseptica M Priest (DAR72350) plants

DAR are national herbarium numbers, NSW Agriculture, Orange. VPRI are Agriculture Victoria collection numbers, Knoxfield Victoria.

48 Table 3.2. Symptoms of disease and the associated disease causing organisms identified on Ixodia achillaeoides.

Symptoms Miscellaneous Identified by Stem damage with Pseudomonas marginalis CHS yellow gumming/exudation Galls on stem of Agrobacterium sp. (Crown Gall) K Ophel-Keller propagated plants in greenhouse Stunting, small leaves, Phytoplasma N Habili apical proliferation

Table 3.3. Symptoms of disease and the associated nematodes identified on Ixodia achillaeoides.

Symptoms Plant parasitic nematodes Identified by and recovered collection number* Root knots and Meloidogyne sp. G Walker galls Meloidogyne hapla G Walker (WINC1027) Pratylenchus sp. G Walker (WINC1040) Soil from root Paralongidorus sp. G Walker (WINC1040) zone of Paratrichodorus sp. G Walker (WINC1040) declining plants Paratylenchus sp. G Walker Tylenchorhynchus sp. G Walker (WINC1040) Hemicycliophora truncata G Walker (WINC1030) Morulaimus sp. G Walker (WINC1030) Radopholoides sp. G Walker (WINC1030) Radopholus sp. G Walker (WINC1030) Rotylenchus sp. G Walker (WINC1030) *WINC are numbers of the nematode collection at Waite Agricultural Research Institute, University of Adelaide, SA

3.3 Pathogenicity studies

Pathogenicity studies were undertaken on potted plants in the greenhouse to confirm that the organisms recovered from the diseased plants were actually causing the disease. This is done by using a pure culture of the organism to infect a healthy plant with the organism. Pathogenicity was confirmed if the same symptoms occured after the inoculation, and the same organism was recovered from the affected plant.

49 3.3.1 Materials and methods

Healthy plants were inoculated with the organisms by various methods (Table 3.4), and after symptoms were observed, plant material was tested as previously described to confirm the presence of the same organism. Tested plants were kept in a growth room with 12 hour day night cycle at 25oC.

3.3.2 Fungi

Leaves were inoculated by spraying them with a 104 – 106 spores/ml suspension of the fungi and enclosing in a plastic bag to provide near 100% humidity for 24 hours to induce infection. Some leaves were pierced with a needle to provide entry points for infection. The control leaves were sprayed with sterile water.

Pure cultures of the fungi were grown on artificial media (PDA or CMA) and after 2 weeks incubation at 22oC, colonies on 5 plates were macerated in 1000ml sterile water. This suspension of hyphae and spores was poured into 4 holes made around the plant and onto the surface of the soil of 8-12 week old potted Ixodia plants, 50ml for 3” pots and 100ml for 6” pots.

Bacteria

Stems were inoculated by dipping a needle into a suspension of 109 bacteria/ml and piercing the stem of the plant with the needle. The control was pierced with a needle dipped in sterile water.

Nematodes

Populations of Meloidogyne sp were multiplied on greenhouse tomato plants. Tomato roots were washed and shaken for 4 min in a sodium hypochlorite solution with 0.5% available chlorine, the resulting suspension passed through nested sieves with 250 and 32µm apertures. Nematode eggs collected on the latter sieve were used to inoculate I. achillaeoides rootlings by adding the suspension to four holes made in the soil around each plant. Nematode-free wash water, obtained by passing the suspension from sodium hypochlorite treated tomato roots through Whatman ® No. 1 filter paper, and containing associated micro-organisms, was added to all pots not inoculated with nematodes.

3.3.3 Results

The pathogenicity of 7 Fungi and 1 nematode species were confirmed (Table 3.4). Other organism were either not tested due to time constraints (nematodes) or because the organisms could not be successfully recultured to obtain a clean and plentiful supply for the testing procedure. Psuedomonas marginalis was recovered from infected stems and was suspected to be an opportunistic bacteria invading after mechanical damage had occurred.

50 Table 3.4 Pathogenicity test methods and results of disease causing organisms recovered from Ixodia.

Organism Test method* Pathogenicity confirmed* Alternaria alternata Leaf inoculation X Stemphyllium sp. - X Capnodium sp. (Sooty mould) - X Oidium sp. (Powdery mildew) Leaf inoculation √ Botrytis cinerea Leaf inoculation √ Fusarium oxysporum Soil inoculation √ Fusarium tabacinum Soil inoculation √ Fusarium acuminatum Soil inoculation X Verticillium dahliae Soil inoculation √ Phytophthora cryptogea Soil inoculation √ Phytophthora erythroseptica Soil inoculation √ Rhizoctonia solani Soil inoculation X Pythium sp. - X Meloidogyne sp. Soil inoculation √ Meloidogyne hapla Soil inoculation √ All other nematodes - X Pseudomonas marginalis Stem inoculation X Agrobacterium sp. (Crown Gall) - X Phytoplasma - X ∗ - not tested ∗ √ pathogenicity confirmed ∗ x pathogenicity not confirmed

3.4 Testing methods of inoculum for Phytophthora.

As previous work with Phytophthora had indicated that applying macerated hyphae to the soil was not the best method of infecting soil and plants with the fungus, an alternative method was evaluated. This involved using vermiculite as the substrate for growing the fungus.

3.4.1 Materials and methods

100ml of V8 juice was mixed with 1g of calcium carbonate and diluted 1:4 with sterile water. 300ml of this solution was added to 600ml of vermiculite and autoclaved for 1 hour. The Phytophthora was then added by cutting squares of agar and mycelia and mixing with the vermiculite. After 2-3 weeks growth at 22oC, 5ml, 25ml or 50ml of the infected vermiculite was then placed around the plant on the soil surface. 22 plants per treatment were used. The plants were flooded for 24 hours on 2 occasions by placing the pots in bags of water to encourage infection by Phytophthora. Inoculated and flooded plants were compared to a flooded not inoculated control, and a not flooded not inoculated control. The results were also

51 compared to those of the same cultivars previously tested using the macerated hyphae inoculation technique.

After 6 weeks, the plant health was rated on a 1-5 scale, where 1= healthy, 2= leaves starting to yellow and die, 3=half dead, 4=>75% of the plant dead to 5 = plant dead. Diseased material from all plants was then tested to ensure the presence of Phytophthora.

3.4.2 Results

Adding 50mls of inoculum gave no better infection rate than 25ml, but both infected or killed more plants than 5ml of inoculum. All levels of vermiculite inoculum infected the plants better than the macerated hyphae, so it was decided in all future testing to apply 25ml of vermiculite inoculum. Flooding plants without inoculation also caused some plant death, indicating that waterlogging may be the cause of plant death even without the added factor of a pathogen present.

Table 3.5. Effect of different methods of inoculation on infection of plants by Phytophthora.

Treatment # plants # plants # with rating # plants with tested dead below 3 Phytophthor a 50ml vermiculite 22 10 4 18 inoculum 25ml vermiculite 22 10 5 20 inoculum 5ml vermiculite 22 5 4 15 inoculum Flooded, no 22 3 13 1 inoculum Not flooded, no 7 0 7 0 inoculum 50ml macerated 22 0 13 12 hyphae

3.5 Cultivar susceptibility

The susceptibility of cultivars from all 3 subspecies of I. achillaeoides to the fungi Verticillium dahliae, Fusarium oxysporum, Phytophthora cryptogea, P. erythroseptica, and Oidium sp. (Powdery mildew) and the species of the nematode Meloiodogyne were tested on potted plants in the greenhouse (Table 3.6). Although not quantified, it was observed that white fly, mites and light brown apple moth also infected most of the plants during testing.

52 3.5.1 Materials and methods

Fungi

Pure cultures of F. oxysporum and V. dahliae were grown on Potato Dextrose Agar (PDA) and used to inoculate potted Ixodia plants as described previously. This was repeated 2 weeks later, and the plant health was assessed weekly for 8-15 weeks after the final inoculation. The plants were rated on a 1-5 scale, where 1=healthy, 2=leaves starting to yellow and die, 3=half dead, 4=>75% of the plant dead to 5 = plant dead.

Inoculation of the Phytophthora sp. was initially carried out using macerated hyphae, but was then repeated using vermiculite inoculum with flooding as previously described. The plant health was assessed after 8-10 weeks as described above.

Plants were inoculated with powdery mildew by dry brushing conidia over the leaves and enclosing all plants in a plastic bag for 24 hours. The level of infection was rated from 1-5, where 1=no infection, 2=1-20%, 3=20-50%, 4=50-75% and 5=>75% leaf area infected.

4 separate tests were undertaken to test all the cultivars, with some cultivars being repeated in each test to provide a continuity. Fusarium was not tested in the 3rd and 4th trials as it was considered to be a less virulent pathogen than the others tested.

Nematodes

One greenhouse experiment was undertaken to test “Hills” and “South East” forms of Ixodia for susceptibility to Meloidogyne sp. This nematode had been recovered from both areas, the symptoms including general wilting, stunting and root galling. Three species of Meloidogyne were tested, M. hapla, M. javanica and M. incognita. Root galling was not severe, particularly from M. hapla, and nematode reproduction was used as a more reliable indicator for susceptibility.

Populations of M. hapla, M. incognita and M. javanica, were multiplied on greenhouse tomato plants. Tomato roots were washed and shaken for 4 min in a sodium hypochlorite solution with 0.5% available chlorine and the resulting suspension passed through nested sieves with 250- & 32-µm apertures. Nematode eggs collected on the latter sieve were used to inoculate I. achillaeoides rootlings by adding the suspension to four holes made in the soil around each plant. Nematode-free wash water, obtained by passing the suspension from NaOCl-treated tomato roots through Whatman® No. 1 filter paper, and containing associated micro- organisms, was added to all pots not inoculated with nematodes.

Fresh root weight and both fresh and dry (72 h at 70°C) shoot weights were measured on all plants. Nematodes were extracted from a 200g subsample of soil per pot on Whitehead and Hemming trays for five days at 22°C and the total number of nematodes per pot counted. Roots were cut into 2cm lengths and transferred to plastic bags for extraction of nematodes by incubation in 3% hydrogen peroxide at 22°C for five days.

Due to a number of plants dying, and low numbers of each cultivar tested, it was difficult to statistically analyse results from each cultivar separately, so those from each district were assessed together as Hills or South East cultivars. One experiment was also not watered for 4 days, so the effect of drought on the infected plants was also observed.

53 3.5.2 Results and discussion

Verticillium dahliae

V. dahliae, isolated from vascular staining in lower and upper stems of wilting plants, was the most destructive of the pathogens tested. Symptoms included chlorosis or desiccation of leaves and death.

All the 58 cultivars tested were susceptible (Table 3.6), with the first symptoms of leaf yellowing and desiccation appearing within 2 weeks and the first infected plants dying within 3 weeks of the second inoculation. All plants that had not died by the final assessment had severe vascular discolouration. V. dahliae was recovered from a random selection of plants exhibiting this staining. Fusarium oxysporum

F. oxysporum caused a slow decline of the plants accompanied by leaf chlorosis. Ixodia plants were inoculated and assessed as described for Verticillium. F oxyporum, was not included in the final 2 tests, as it was considered to be less pathogenic than Verticillium or Phytophthora.

All 25 cultivars of Ixodia tested were susceptible (Table 3.6), but plant death occurred in only a few of the plants, less than half the number of those that died when inoculated with V. dahliae. Most of the inoculated plants had vascular staining, from which F. oxysporum was successfully recovered from a random selection.

Phytophthora sp.

P. erythroseptica and P. cryptogea were isolated from stem cankers and rotted roots of chlorotic plants. Root rotting was severe and cankers often extended into the lower branches.

All 53 cultivars tested were susceptible to both species when grown in soil artificially infested with Phytophthora (Table 3.6). Some of the cultivars that had not been affected in the first 2 trials were retested and found to be susceptible using the vermiculite inoculum. Phytophthora was recovered from a random selection of affected plants in each trial, and from soil baits from pots of dead plants.

To determine if the geographic origin of the cultivars played a role in relative susceptibility to these phytophthora species, one-way AOV was run on ranked?? Ratings, grouping varieties from 4 major geographic classes: Adelaide Hills (H), Nelson Area, Vic (N), Grampians, Vic (G) and Kangaroo Island (KI) and Mt. Richmond,Vic. (MR).

54 The results of means for P. cryptogea susceptibility are:

Geo Mean Grampians 4.7500 a

Nelson 4.0455 b

Hills 3.3333 c

Kangaroo Island 3.1667c

The results of means for P erythroseptica susceptibility are:

Geo Mean Grampians 4.7500 a

Nelson 4.0909ab

Hills 3.7500bc

Kangaroo Island 3.5000bc

Mt Richmond 2.5000c

Powdery mildew

Of the 61 cultivars tested, all but 7 were susceptible (Table 3.6).

Table 3.6. Susceptibility of cultivars to the common fungal pathogens. Geographical classes are as used in Part II.

Assess. Geo. Verticilliu Fusarium Phytophthora Phytophthora Powdery oxysporum cryptogea erythroseptica mildew # m dahliae

18 G 5 5 5 5 19 G 3 5 5 3 20 G 3 3 3 4 5 49 G 5 5 5 2 50 G 5 5 5 2 67 G 4 5 4 4 312 G 5 5 5 2 316 G 5 5 5 1 151 H 5 3 5 3 5 2 H 5 3 3 4 3 5 H 5 3 3 5 2 6 H 4 3 2 2 1 7 H 5 4 3 2 1

55 36 H 3 4 5 2 37 H 2 39 H 4 4 3 2 1 40 H 4 5 4 4 4 42 H 4 3 3 4 2 47 H 3 3 3 4 2 48 H 5 4 5 2 115 H 3 3 5 2 541 KI 5 4 5 46 KI 3 3 3 3 62 KI 2 3 3 4 1 510 KI 3 3 3 4 2 516 KI 4 5 4 2 521 KI 4 4 2 2 4 527 KI 3 2 3 4 5 17 MR 5 3 3 2 4 721 MR 5 5 3 3 4 65 N 4 5 5 3 21 N 3 4 5 2 25 N 3 3 26 N 4 3 27 N 5 2 3 235 N 4 3 236 N 5 4 3 N 5 3 5 4 4 8 N 4 4 3 2 9 N 4 5 4 3 10 N 4 5 4 2 11 N 5 5 5 3 14 N 4 4 4 2 15 N 4 4 4 2 16 N 4 3 3 3 1 22 N 4 3 3 2 23 N 5 4 3 3 24 N 5 3 3 1 25 N 5 4 4 2 26 N 4 2 3 2 27 N 5 4 4 2 29 N 5 4 5 2 30 N 5 3 4 2 31 N 5 5 5 3 34 N 3 4 5 5 348 N 1 5 5 3 711 N 5 4 5 3 43 SE 5 3 4 4 5 13 SE 5 4 5 32 DC 4 3 5 5 3 644 E 3 5 4 4

56

Nematodes

All plants tested were susceptible to the nematodes, however the susceptibility varied between Ixodia populations, and between Meloidogyne spp. within a population (Table 3.7). There was a much lower number of juveniles of either M. incognita or M. hapla found in the Hills cultivars compared to the South East cultivars.

Root and shoot biomass were consistently lower in inoculated plants of both populations, however, only M. incognita and M. javanica significantly reduced shoot growth (Table 3.7). M. javanica and M. hapla significantly reduced root growth of the South East population. These growth reductions indicate that these populations were intolerant of these particular nematodes.

When plants were infected with Meloidogyne , the plants (particularly Hills cultivars infected with M. incognita and M. javanica) were much less drought tolerant. Two of the Hills cultivars, LS311T and FRS95(2) appeared to be the least tolerant to drought and prior infection with Meloidogyne could cause severe losses in these lines under drought conditions.

Table 3.7. Resistance and tolerance of 2 I. achillaeoides populations to 3 Meloidogyne spp. in greenhouse experiments.

Meloidogyne % dead Plant weight (g) Number of juveniles spp. plants raw and (transformed data ln(x+1) ) Dry shoots Fresh roots Roots Roots + soil South East cultivars Uninoculated 0 2.3 1.3 0 0 0 0 M. incognita 10 1.7 1.1 2097 (6.97) 2177 (7.22) M. javanica 0 1.4 0.7 1114 (6.32) 1208 (6.46) M. hapla 0 2.0 0.8 542 (4.90) 553 (5.04) LSD (P=0.05) - 0.4 0.4 (1.42) (1.20) Hills cultivars Uninoculated 0 4.1 1.9 0 0 0 0 M. incognita 30 2.2 1.0 521 (5.85) 523 (5.83) M. javanica 40 3.1 1.3 362 (4.37) 409 (4.69) M. hapla 0 3.7 1.3 559 (6.06) 681 (6.34) LSD (P=0.05) - 0.5 NS (1.18) (1.08)

57 3.6 Nematode and Verticillium Disease complex

It had been observed in commercial plantings that Ixodia infected with both Verticillium and Meloidogyne spp caused more severe wilting and plant death than plants infected with only one of the pathogens. A greenhouse trial was established to examine the effect of infection by one or both of these pathogens.

3.6.1 Materials and methods

V. dahliae was grown on potato-dextrose agar (PDA) for 20 days at 22°C. The fungus and agar from five 90mm diameter petri plates were dispersed in 1 L deionised water in a Waring blender. I. achillaeoides plants were inoculated with 10 ml of this suspension containing 22 × 107 conidia and 7.8 × 103 microsclerotia per ml by adding the suspension to four holes made in the soil around each plant. Plants not inoculated with V. dahliae received 10 ml of a suspension made up from five uninoculated PDA plates dispersed in 1 L deionised water.

Vascular tissue in stems of plants were examined for discolouration, and stem tissue from all plants was incubated on PDA amended with streptomycin sulphate (200 mg/L) for 14 days at 22°C to isolate V. dahliae.

3.6.2 Results and discussion

Symptoms similar to those seen in the field (severe wilting and death of plants) were only observed in this greenhouse experiment in plants that were inoculated with both V. dahliae and Meloidogyne spp. Although V. dahliae reduced root and shoot growth, it did not kill plants except when plants were also inoculated with Meloidogyne spp. (Table 3.8). Severe staining of vascular tissue of the basal stem, associated with infection by V. dahliae, was also only observed in dying plants infected with both pathogens. This test used a lower inoculum dose than the cultivar susceptibility tests, which may explain the much higher disease levels observed in the latter tests.

The disease complex between these two pathogens was not reflected in more severe reductions in plant growth following inoculation with both pathogens compared with either alone (Table 3.8). However, this analysis excluded the plants which had already died during the course of the experiment and, if included, these would have reduced substantially the mean weights for plants inoculated with both pathogens. Root growth of plants inoculated with V. dahliae alone was nevertheless significantly lower than that of plants inoculated with both pathogens (Table 3.8).

58 Table 3.8. Effects of V. dahliae and Meloidogyne spp., alone and in combination, on growth and longevity of I. achillaeoides (South East population) plants in the greenhouse.

Treatment Plant weight (g) % dead or Number of Meloidogyne dying juveniles (± S.E.) Dry Fresh plants Roots Roots + soil shoots roots Uninoculated 3.0 2.1 0 0 0 V. dahliae 2.0 1.0 0 0 0 Meloidogyne spp 2.3 1.8 0 325 ± 97 400 ± 106 V. dahliae and M. 2.5* 1.9* 40 480 ± 225 536 ± 229 spp LSD (P=0.05) 0.6 0.7 - - - * dead plants excluded from plant weight data

3.7 Control of Powdery mildew

Two experiments were undertaken in the greenhouse using potted Ixodia plants severely infected with powdery mildew to test the efficacy of various fungicides against the disease.

3.7.1 Materials and methods

Inoculation

Leaves infected with powdery mildew were initially collected from a planting in the Adelaide Hills. Spores were brushed from the infected leaves onto the leaves of potted plants and covered with a plastic bag for 24 hours to provide high humidity. After several plants were heavily infected, these were used to inoculate more plants. However after the spores had been brushed onto the plants, one of the infected plants was enclosed in the bag with several inoculated ones.

Experiment 1.

10 plants per treatments were tested using the fungicides and rates outlined in Table 3.9. The 10 plants were chosen from the inoculated plants so all treatments had the same cultivars with similar levels of infection for each treatment. All chemicals were mixed with water and sprayed to run off using a hand held atomiser, applying between 300 and 400ml per treatment. 3 occasions at 10 day intervals. A wetter was added to all fungicide treatments.

Plants were watered without wetting the leaves. All treatments were applied in a separate area and allowed to dry before being returned to the greenhouse to prevent contamination of other treatments from the chemicals.

Plants were assessed for the percent leaf area covered with mildew on 6 occasions, from prior to the initial spray to 34 days after the third spray was applied.

59 Table 3.9. Fungicides and rates used for powdery mildew control, Experiment 1. Treatment Active ingredient Rate of product Control Water Bayfidan 250 EC® (+citowett®) 250g/Kg Triadimenol 0.1ml/L (+0.1ml/L) Ampol DC-Tron ® (+citowett®) 839 g/L Petroleum oil 10ml/L (+0.1ml/L) Kumulus® (+citowett®) 800g/Kg Sulphur 3g/L (+0.1ml/L) Citowett® 1000g/L Alkylaryl polyglycol ether 0.4 ml/L

Experiment 2.

10 plants per treatment were tested using the fungicides and rates outlined in Table 3.10. Treatments were applied to the plants as described previously on 5 occasions at 7-10 day intervals. Plants were assessed as previously described on 8 occasions from prior to the first spray to 3 weeks after the last spray was applied. The amount of mildew on the leaves was rated from 0-4, where 0=no disease, 1=1-25% , 2=26-50%, 3=51-75% and 4=76-100% of leaf area covered with mildew.

Table 3.10. Fungicides and rates used for powdery mildew control, Experiment 1.

Treatment Active ingredient Rate of product Control Water Kumulus® 800g/Kg sulphur 3g/L Citowett® 1000g/L alkylaryl polyglycol ether 0.1ml/L Ampol DC-Tron® 839 g/L petroleum oil 10ml/L Baking soda 10g/L Topas 100EC® 100g/L penconazole 1.5ml/L Bayleton® 50g/Kg triadimefon 0.65g/L Rotam Flosul SC® 800g/L sulphur 2ml/L Rubigan 120 EC® 625g/L fenarimol 0.2ml/L Benlate® 500g/Kg benomyl 0.5g/L

3.7.2 Results and discussion

Experiment 1

All treatments but Citowett® significantly reduced the level of mildew compared to the control (Fig 3.1). Kumulus® was the most effective of the treatments, completely controlling the mildew within 14 days from the first spray, and no regrowth of the fungus was observed up to 38 days after the third and final application (Fig 3.1). Bayfidan® and Ampol DC Tron® were also effective in controlling the fungus when sprayed, but the small amount not controlled with the fungicide caused reinfection of the leaves after the final spray.

60 Fig 3.1. Level of mildew on Ixodia plants with and without fungicides, Experiment 1.

100 90 Sprays applied Control® 80 70 Citowett®

60 L.S.D. (P=0.05) = 31 50

40 Bayfidan® Kumulus® 30

% leaf area with mildew Mean 20 Ampol DC Tron® 10

0 0 9 14 21 29 59 Days from first spray

Experiment 2

All treatments significantly reduced the level of mildew at the final measurement compared to the untreated control (Fig 3.2). Kumulus®, Rotam® sulphur, Bayleton® and Topas® were the most effective, with no regrowth of the fungus up to 21 days after the final application. Citowett® was again the least effective, while the other treatments, although reducing the level of mildew, did not prevent re-infection once spraying was ceased (Fig 3.3).

Fig 3.2. Level of mildew on Ixodia plants with and without fungicides, Experiment 2.

4.5 Spray dates

4

3.5 Control Kumulus Citowett DC Tron Plus 3 Baking Soda Topas 2.5 Bayleton Rotam 2 Rubigan Benlate

1.5

1 Mean Powdery Mildew Rating 0.5

0 24/07/1998 31/07/1998 10/08/1998 17/08/1998 24/08/1998 31/08/1998 7/09/1998 14/09/1998

Assessment date

61 Fig 3.3. Level of powdery mildew on Ixodia 1 and 3 weeks after the application of the 5th and final spray of various fungicides.

4.5

g 4 1 week after final spray 3.5 3 weeks after the final spray 3 2.5 2 1.5 1

Mean powdery mildew ratin 0.5 0

te n s s wett oda eton ulu o l otam Control g S Topa R Cit Benla Rubiga Bay Kum Tron Plus Bakin DC Treatments

3.8 Control of Phytophthora

2 greenhouse and 1 field trial were undertaken to evaluate the efficacy of registered fungicides to control Phytophthora on Ixodia. Phytophthora cryptogea and P. erythroseptica previously isolated from Ixodia were used in all trials to make vermiculate inoculum as previously described. The vermiculate from the 2 species was combined 1:1 and used together to infect the plants.

3.9 Greenhouse experiments

The 2 greenhouse experiments were undertaken on several susceptible cultivars, using the same number of each cultivar in each treatment.

3.9.1 Materials and methods

Potted Ixodia were inoculated with 25ml of vermiculite as described previously. Treatments and rates applied are outlined in Table 3.11 (Expt 1) and Table 3.12 (Expt 2). All pots were flooded twice before the “after inoculation” treatments were applied. Between 15 and 20 plants per treatment were used, with foliar sprays using 300-350 ml/treatment. Plant health was assessed 6-16 weeks after inoculation using the 1-5 rating system as previously described. Soil in the pots was mixed and baited, and diseased material from plants was tested to detect the presence of Phytophthora.

62 Table 3.11. Treatments and rates applied to potted Ixodia to test efficacy against Phytophthora, greenhouse experiment 1.

Treatment Application time and method Rate of product Control – inoculated Foli-R-Fos® Foliar spray before inoculation 10ml/L Foli-R-Fos® Foliar spray before and after inoculation 10ml/L Foli-R-Fos® Foliar spray after inoculation 10ml/L Acrobat Soil drench after inoculation 50g/L, 20ml/pot Ridomil® granules Soil treatment after inoculation 1g/pot

Table 3.12. Treatments and rates applied to potted Ixodia to test efficacy against Phytophthora, greenhouse experiment 2.

Treatment Application time and method Rate of product Control (flooded)* Control (not flooded)* Foli-R-Fos® Foliar spray before inoculation 10ml/L Foli-R-Fos® Foliar spray before and after inoculation 10ml/L Foli-R-Fos® Foliar spray after inoculation 10ml/L Ridomil® granules Soil treatment after inoculation 1g/pot

*controls were not inoculated

3.9.2 Results and discussion

None of the treatments controlled the Phytophthora, the lowest plant death occurring with Foli-R-Fos® applied before and after inoculation (Table 3.13, 14). With the flooding of the pots, the fungus would spread and establish in the soil and possibly start to infect the plant before the treatment was applied. Spraying Foli-R-Fos® before inoculation would have provided some level of protection for the plant. All plants tested were infected with the fungus, even if they had not yet died (Table 3.13). Ridomil® granules and the pre and post inoculation application of Foli-R-Fos® both reduced the level of fungus in the soil, as in some pots the fungus could not be detected by the baits (Table 3.14).

There was also some plant death in the not inoculated controls (Table 3.14), especially those that were flooded. This supports the earlier finding that waterlogging alone would cause dead of Ixodia.

63 Table 3.13. Effect of treatments on plant death at various times after inoculation with Phytophthora, greenhouse experiment 1.

Treatment No plants % plants dead % plants with tested 13 weeks 16 weeks Phytophthora Control - 15 53 87 100 inoculated Foli-R-Fos® pre 15 27 60 100 inoculation only Foli-R-Fos® pre 15 33 47 100 and post inoculation Foli-R-Fos® post 15 53 60 100 inoculation only Acrobat 15 67 93 100 Ridomil® granules 15 47 60 100

Table 3.14. Effect of treatments on plant death at various times after inoculation with Phytophthora, greenhouse experiment 2.

Treatment No plants % plants dead soil baits with tested 22 days 41 days Phytophthora %/no. tested Control not 15 0 7 0 / 15 flooded* Control flooded* 20 10 25 0 / 11 Ridomil® granules 17 0 76 67 / 12 Foli-R-Fos® pre 17 12 100 100 / 14 inoculation only Foli-R-Fos® pre 17 12 65 93 / 14 and post inoculation Foli-R-Fos® post 17 24 88 100 / 15 inoculation only

*These treatments were not inoculated.

3.10 Field phytophthora inoculation trial

The field experiment was undertaken in a small plot at the Lenswood Research Centre that had previously been used for potato pink rot trials, and was infected with P. erythroseptica. Ixodia were planted on 3rd December 1998 in 12 rows of 18 plants, and allowed to establish for almost 2 months before being inoculated on 24th Feb 1999. Any plants that died in this time were replaced with healthy plants.

64 3.10.1 Materials and methods.

50ml of vermiculite inoculum was placed in a shallow trench around the base of the plant and covered with a layer of soil. The plants were waterlogged on at least 2 occasions after inoculation by watering with overhead irrigation for >6 hours.

4 replicates of each treatment were laid out in a randomised block, the treatments and rates applied as in Table 3.15. After the Ridomil® granules were applied, the plot was irrigated for 2-3 hours to leach the chemical into the soil profile. The foliar sprays of Ridomil® or Foli-R- Fos® were applied the following day.

The number of dead plants was assessed on 6th July and 28th Sept., 1999, and all dead plants were tested for the presence of Phytophthora.

A second inoculation was applied on 5th October 1999, with the same treatment regime applied. Plants were reassessed on 8th March and 1st June 2000.

Table 3.15. Treatments and rates applied to Ixodia to test efficacy against Phytophthora in a field experiment.

Treatment Application time and method Rate of product Control not inoculated Control inoculated Ridomil® granules 25G Soil treatment 6 days after 50g/m2 (25g/Kg) inoculation Ridomil Gold® MZ Foliar spray 1, 4, 9 weeks after 2.5g/L (40g/Kg) inoculation Foli-R-Fos® (200g/L) Foliar spray 7 days before 10ml/L inoculation, 1, 4, 9 weeks after inoculation Foli-R-Fos® (200g/L) Foliar spray 1, 4, 9 weeks after 10ml/L inoculation

3.10.2 Results and discussion

The first assessment and second assessments, undertaken between 4 and 6 months after the first inoculation, showed Ridomil® Gold to be the only treatment to effectively reduce the number of dead plants (Table 3.16). However by March the following year, 6 months after the second inoculation, Ridomil® granules and Foli-R-Fos® applied before and after inoculation had more plants still alive.

65 Table 3.16. Effect of treatments on plant death at various times after inoculation with Phytophthora, field experiment.

Treatment Total plants dead (%) Total plants still 6/7/99 28/9/99 8/3/00 alive (%) 8/3/00 Control not inoculated 22 22 41 59 Control inoculated 16 28 34 66 Ridomil® granules 13 28 28 72 Ridomil® Gold 3 9 41 59 Foli-R-Fos® pre and post 19 22 28 72 inoculation Foli-R-Fos® post inoculation only 16 22 47 53

By the final assessment, the Foli-R-Fos® foliar spray applied before and after inoculation was the most effective treatments, with 66% of the plants still healthy (Table 3.17).

Table 3.17. Effect of treatments on plant death on 1st June 2000, 9 months after second inoculation with Phytophthora, field experiment.

Treatment Total Total Total plants plants dead plants still healthy (%) dying (%) (%) Control Not Inoculated 41 18 41 Control Inoculated 34 35 31 Ridomil® granules 28 18 44 Ridomil Gold® 41 12 47 Foli-R-Fos® pre and post 28 6 66 inoculation Foli-R-Fos® post inoculation only 47 12 41

Phytophthora was recovered from at least 50% of the soil baits from dead plants in inoculated treatments (Table 3.18). Phytophthora was also recovered from all soil tests on plants from the inoculated control, and 2 of the 6 from the not inoculated control. This was not unexpected, as the plot area had been previously infected with P. erythroseptica, and there would also have been some movement of the fungus in the soil from the inoculated plots.

66 Table 3.18. Result of testing for Phytophthora from dead and dying plants, field experiment. Treatment Sample 8/3/00 Sample 1/6/00 No dead No soil No root No dying No soil plants baits +ve isolations plants baits +ve tested +ve tested Control 6 2 0 5 4 Not Inoculated Control 5 5 1 4 4 Inoculated Ridomil® 0 9 6 granules Ridomil Gold® 10 5 3 2 2 Foli-R-Fos® pre 2 2 0 2 2 and post inoculation Foli-R-Fos® 8 4 3 2 1 post inoculation only

These results indicate that none of the chemicals currently registered will eradicate the fungus from the soil. Continual treatment would be necessary to keep plants healthy for the useful life of the plant. Foli-R-Fos® appeared to have the most residual effect of the fungicides tested, but was more effective when applied before infection as well as after. Ridomil Gold® was very effective initially, but did not have any long term benefit. Ridomil® granules were effective for longer, but the residual effect did not appear to be as long as with the Foli-R- Fos®. The most effective spray regime would therefore seem to be a combination of these fungicides, perhaps using Ridomil® Gold or granules initially, with maintenance sprays of Foli-R-Fos®.

67 4. Commercialisation

4.1 Introduction

The objective of the assessment and development program described in the previous sections of this report has been to recommend and deliver improved varieties of Ixodia achillaeoides to the Australian floriculture industry. The chief outcome of this program is that varieties are available for commercialisation that have known flowering, cultivation and postharvest characteristics and which have been assessed by growers and wholesalers for market performance. Performance standards have been determined by the continued evaluation of selected varieties in replicated field trials supported by laboratory and greenhouse investigations. Meetings with growers groups and trial shipments have been used to evaluate and predict market performance.

Superior varieties have now been identified for commercialisation and in partnership with RIRDC and industry, the best methods of protecting and disseminating varieties for the benefit of industry will be developed.

4.1.1 Background, relevance and potential benefits

The provision of selected varieties of ixodia with improved yield, appearance and performance and known flowering and cultural requirements will provide the following outcomes:

• Producers can make informed choices on varieties to grow with known performance standards which best suit their sites, production systems and targeted markets. • Major disease problems of ixodia have be identified and selections tolerant to these diseases evaluated and incorporated into the improvement program. Disease control measures have been evaluated and recommendations made for commercial production. • Based on the results of this program, producers have the technical information required to operate a quality assurance program . • An understanding of the economics of ixodia production based on high quality, reliable clones would lead to a confident promotion and growth of further plantings to help meet market demand. Ixodia will become a profitable crop to produce.

4.1.2 Economic benefits

The results of this project will conservatively realise a 4-5 fold increase in the value of production of Ixodia to a level of $2 million two years after initial adoption of new varieties. The flow of improved varieties would see this value continue to increase over the following five years. This project has the potential to benefit fresh and dried flower producers in southern and western Australia where ixodia is growing as their primary crop or as an alternative line. Ixodia is not currently exported from other overseas countries with active flower export industries. With proper protection in the introduction of new varieties, Australian producers will directly benefit from the results of this program. Potential exists for the marketing of pot plant lines to overseas markets which would result in financial benefits which would flow back to wildflower research through RIRDC and SARDI.

68 4.2 Research strategies and methodology

Progeny from field collections and controlled crosses performed since 1997 have been and will continue to be evaluated in greenhouse trials and promising individuals ’fast tracked’ in field evaluations. The potential exists for new varieties to be slowly introduced to industry over several years to stimulate interest and continued development of the crop.

Mother block plantings of all superior varieties have been established at Lenswood Centre and will be maintained to serve as stock plants for multiplication of selected varieties for commercialisation. Source material for all varieties evaluated and collected in the program (>300) will be maintained in containers and field for reference and future breeding programs until given to a commercial partner.

Leading Australian floriculture and plant development companies have been contacted and initial discussions held in relation to commercialisation of both cut flower and pot plant lines. Issues such as protection of varieties, overseas agencies, propagation methods and marketing in Australia and overseas have been discussed. There is excellent potential for an Australian company to successfully commercialise these lines.

4.3 Communications/adoption/commercialisation strategy

It is suggested that flower producers in SA, Victoria and WA primarily be targeted with results of research, information on new varieties, and market data.

In consultation with SARDI, RIRDC and industry representatives a commercialisation strategy should be developed with the successful commercialisation partner. This will allow for early benefits to the Australian cutflower industry in building up production levels to develop and sustain domestic export markets. The commercialisation partner will propagate, promote and market new varieties. PBR protection is recommended for outstanding varieties such as double steriles, forms for protected cultivation or pot plant lines.

Identification of initial varieties for commercialisation has been made and is included in a confidential report to RIRDC. It is recommended that the new ixodia varieties released for cut flower production be marketed under the identified trademark of Southern Star Series. This may include some PBR protected varieties as well as some that are not registered. Recommendations will be made that some grower contributed varieties be included in this commercialisation program. Agreement has been made in principle with these growers to participate in discussions with the selected commercial partner, concerning the inclusion of their varieties in the program.

69 5. Appendices

5.1 Appendix I.: Records of herbaria searches for locations of Ixodia ahillaeoides ssp.alata 1 collections. 1 unless otherwise noted

SOUTH AUSTRALIAN HERBARIUM RECORDS

LOCATION COLLECTION COMMENTS Southern Mt Lofty Ranges DATE

McHarg Creek. Bull Creek Range. 24.6.1989 Small flower. Common.

Finniss. Ca. 55km S.S.E. of Adelaide. 1.2.1926 Ca. 15km W. of Milang.

Cox's Scrub National Park. 18.3.72 Flowers up stem. Near Mt. Observation. Ca. 55km S.S.E. of Adelaide.

Kyeemain National Park. 12.3.77 Long narrow leaf. Could be some interesting Sth boundary. Kyeemain ca. 45km ones in this area. S.S.E. of Adelaide.

Mt. Compass 26.1.41

Kuitpo 15.2.53

Millbrook Reservoir. 1.2.76 Long fine leaf. Small terminal flower. ( 34 48' S. 138 49' E. )

Deep Creek 22.1.81

Kangaroo Island

Cape du Cauedic. Near lighthouse. 11.11.58 Bud

Cape du Cauedic Lighthouse. 27.12.57 Just open.

20yds N. Remarkables Rocks. 6.1.66 Prostrate shrub. Roundabout Cape du Cauedic

Cape du Cauedic. 7.12.80 T.Reichstein 3110 . Photo.

Cape du Cauedic. Top of 100ft cliff E. of 0.12.68 Flowers not quite open. the Cape.

Remarkable Rocks. Ca. 5km E. of Cape 13.1.62 6 - 9" high x 18" wide. du Cauedic.

Willson River Mouth. Mouth Flat. 17.11.89 Buds. ( 35 50' 20" S. 137 57' 35" E. )

Willson's River. 23.1.43 Large flowers.

70 Ca. 3km S. Antechamber Bay 23.1.43 Bigger flower.

Cape Borda. 4.3.26 Flowers 4 - 5mm over 100 in head. Head 60mm across.

Penneshaw. Hillside above beach. 4.3.77 Flowers large. 60cm high shrub.

Penneshaw Rubbish Dump. 4.3.77 Longer broader leaves.

4.5mls E. of Penneshaw Rubbish Dump on 4.3.77 Longer broader leaves. road to Cape Willoughby.

7.5km E. of Penneshaw Rubbish Dump, 4.3.77 Long leaves. Big flowers. Dudley Peninsula.

False Cape. Ca. 3.5km N.E. of S. coast on 14.12.86 Buds. fire access track. Dudley Peninsula.

Cape Willoughby 9.3.1884

Parndana. 0.3.84 Large flowers

Stunsail Boom River area. 14.11.71 Plant 5ft high. Flowers larger than usual with fewer heads.

Hilltop 1.5mls S. of Nth coast road along 9.3.77 Small flowers. Longer leaves. road between Stokes Bay & Parndana.

South-east

Noolook Forest. Bagdad Scrub. 7.1.85 Smallish flowers, compact head with fairly large number of flowers.

Douglas Point. W. of Pt. MacDonnell 25.3.78 Ixodia achilleaoides ssp arenicola

Carpenters Rocks " ( 37 55' S. 140 24' E. )

Yorke Peninsula Compact dense plants. Cape Spencer. 19.3.77 Flowers old. (Prostrate). Dwarf compact.

1.8km W. of Marion Bay. 19.3.77

Innes National Park. Cliffs above wreck 14.5.84 Common. Forming rounded hummocks. of Ethel . ( 35 16' S. 136 50' E. )

Innes National Park. N. tip Gym Beach. 10.10.74 Just in bud.

Stenhouse Bay. 13.10.63 Bud.

Pondalowie Bay. Ca. 10km N.W. of 0.12.1928 Centre of flower closed. Compact plant. Cape Spencer. Between Corney Point & Cape Spencer.

Warrenben Conservation Park.Southern 30.1.84 Full flower.Small flowers, strong bush.

71 boundary near Stone Wall. Flowers all the way up the stem. ( 35 07' S. 137 00' E. )

Marion Bay. 18.1.73 Small domed shrub.

14.2km N.E. of Marion Bay on 19.3.77 Warooka Road.

Between Marion & Stenhouse Bay. 7.2.71 Plant aprox. 4m across & 2 1/2m high.

Flinders Ranges Ixodia flindersica

Mt Hack. Ca. 50km S.E. of Liegh 15.5.77 Flowers look double. Creek

Mt. Serle. Ca. 45km E. of Liegh 8.4.77 Creek. Creek beds, lower slopes.

Mt. Serle. S.E. slopes. 8.4.77

Near Mt. Serle. Ca. 40km E.S.E. of 13.10.62 Flowers open. Liegh Creek.

Mt. Rose. 25km on Mt. Serle track. 2.11.89 Just opening. Near summit S.W. facing rock wall.

Mt. Rose. N. Mt. Serle H.S. 30.7.80 ( 30 19' S. , 138 56' E. )

Mudlapena Gap. Angepena Station 31.7.80 Flowers gone. ( 30 36' S , 138 48' E. )

Freeling Heights." Yudnamutana." 21.7.80 Leaves much broader from this location. 1:50,000 map 6737-1 Grid ref. 457-642 Common. ( 30 09' S., 139 23' E. )

Freeling Heights 19.7.80 Flowers gone. Common.

Mt. Livingstone Cave. 20km N. 7.3.76 Flowers mature.Only 3 plants. Flowers off Freeling Heights. ( 30 09' S., 139 23' E ) white.

S. of Bolla Bollana Creek. 27.10.80 Flowers just open. ( 30 24 S. 139 13 E . )

( 30 24' 30" S. 139 11' 00" E. ) 30.11.80 Flowers just open.

Balcanoona Range. Gammon Ranges 29.9.84 Buds. Plants infrequent. National Park.( 30 33' S. 138 10' E. )

Gammon Range National Park. 2.10.81 Bud. Not common. Near Wildflower Creek.

Mt. McKinlay. Gammon Range. 21.7.80 Flowers gone. ( 30 31' S. 139 06' E. )

72 Mt. Andra. N.W. Narrina H.S. 3.6.80 Very mature flowers. Common. ( 30 56' S. 138 54' E. )

Mt. Uro Range. Wankowoodna Gap 3.6.80 N. of Narrina H.S.

Cocks Comb. N. of Narrina H.S. 3.6.80 Flowers gone. Common.

Ann Hill. Adjacent to Point Well H.S. 3.6.80 No flowers. Common. S. of Narrina H.S.

Mt. Tilley. W. Narrina H.S. 1.6.80 Flowers gone. Uncommon.

Mt. Patawerta. N. of Blinman. 1.6.80 Flowers old. Common.

Balancing Rock Gorge. Ca. 4km E.N.E. 16.7.80 Broad leaf. Flowers gone. Yudnamutana Mines. ( 30 10' S. 139 17' E. )

Yudnamutana Waterfall. Ca. 1 1/2km 18.7.80 Common. S.S.E. Yudnamutana Mines.

Gorge E. Patsy Springs. Depot Springs 28.7.80 Common. H.S. ( 30 31' S. 138 52' E. )

Mt. Shanahan Mine. 5.12.81 Slightly broader leaf. 1 plant seen. ( 30 05' S. 139 23' E. )

Victoria

S.W. 20km S.W. of Edenhope. 7.3.84 Roadside. Fine leaf. Flowers have several Grid. D. 10 rows of petals. Almost looks like BB-AC2

Anglesea 1.2.46 Fine leaf.

Torquay 0.1.1926 Multy headed, fine short leaves.

Portland Road 18mls E. of Nelson. 6.11.67 Flowers just open. Several rows of Coastal sandhill. notched petals.

Nelson. 1km along Beach Road from 9.12.90 About ready to harvest. corner of Main Rd. S. side of Rd just below Southern most parking bay.

Near Donovans Landing. Glenelg River. 20.2.81 Lots of flowers.

Glenelg River estuary. Cliffs overlooking 17.1.85 Mud Lake.

Portland. Coast near Alcoa aluminium 20.2.81 Ixodia achilleaoides ssp arenicola smelter.

73 VICTORIAN HERBARIUM. LOCATION COLLECTION COMMENTS DATE

12mls S.W. of Portland. Along 22.1.69 Bushes 2' tall . Abundant along coast. coast at Cape Nelson.

Dunes of Bridgewater Bay. Portland 31.7.80 38 22' S , 141 29' E

Portland. Gorae West. 0.12.45 Big flowers.

Scenic Rd. S. of Portland. 15.11.83 Big flowers , terminal. 38 12' S. , 141 36' E.

Lower Bridgewater. 17.10.51 Pink form. 10 - 12mm flw. Leaves 4mm diam. 2cm long terminal , large no. of flowers.

Nth boundary of Mt. Richmond 22.10.60 Fl. not quite open. Nat. Pk.

Between Gorae West & 16.10.52 4 - 10' high. Fl. not quite open. Mt. Richmond.

Roseneath Flora Res. 14.3.84 Small centre,several rows of notched petals.

Serra Range E. of Minnaratwa 24.5.68 Thin 1 - 1.5cm stems. Very small flowers. 39.1mls SSW of Stawell

Otway Plain, N. side of Point Addis 28.3.93 Full bloom. Thin 2cm leaves. Browning from Rd. 2km from Anglesea Rd. base.

Point Addis coastal Res. 6km ENE 19.1.79 of Anglesea.

Anglesea Flora Res. 13.3.82 Fine leaf , terminal flower.

Anglesea. Near the intersection of 17.2.85 the Anglesea Rd. & Forest Rd.

Great Ocean Rd. Anglesea. 1.5.43 Heathland. ( Towards Torquay. )

Portland, Bridgewater. 0.1.45 Ixodia achilleaoides ssp arenicola

South Australia

Cape Wiles near Pt Lincoln 6.3.60 1cm leaves thick, flowers 7mm.

Memory Cove. Eyre Peninsula. 19.2.88 Flowers small, 5mm. Thin leaves. Compact 34 59' S. , 135 59' E. plant.

5km down Cape Hart Rd. from 13.4.91 not very big flower heads. intersection.35 52' S. , 138 02' E.

74 5.2 APPENDIX II: Summary of harvest dates of all Ixodia varieties assessed at the Lenswood site, grouped by geographic origin.

Geographic Year 1 Year 2 Year 3 Year 4 Average Variety Region 96/97 97/98 98/99 99/00 Harvest Date 426 A 5.1.00 5/1 427 A 5.1.00 5/1 428 A 5.1.00 5/1 413 A 9.2.00 9/2 411 A 9.2.00 9/2 415 A 15.2.00 15/2 416 A 15.2.00 15/2 47 DC 19.12.96 30.12.97 23.12.98 24/12 32 DC 30.12.96 31.12.97 23.12.98 5.1.00 30/12 33 DC 31.1.97 20.1.98 19.1.99 31.1.00 25/1 160 DC 5.1.00 5/1 161 DC 7.1.00 7/1 166 DC 26.1.00. 26/1 64 E 7.1.97 23.12.98 10.12.99 23/12 615 E 13.1.99 13/1 611 E 3.2.99 3/2 14 G 7.12.97 1.12.98 4/12 15 G 17.12.96 4.12.97 1.12.98 7/12 28 G 31.12.97 29.12.98 5.1.00 1/1 319 G 5.1.00 5/1 20 G 31.1.97 28.1.98 19.1.99 25.11.99 10/1 331 G 19.1.99 19/1 352 G 19.1.00 19/1 72 G 28.1.99 28/1 49 G 4.2.98 3.2.99 3/2 76 G 3.2.99 3/2 312 G 3.2.99 3/2 313 G 3.2.99 3/2 316 G 3.2.99 3/2 332 G 3.2.99 3/2 50 G 10.2.98 3.2.99 6/2 67 G 11.2.00 11/2 18 G 26.2.97 10.2.98 3.2.99 13/2 19 G 26.2.97 17.2.98 9.2.99 17/2 160 H 5.1.00 5/1 40 H 14.1.97 16.1.98 13.1.99 31.1.00 18/1 7 H 22.1.97 28.1.98 13.1.99 28.1.00 22/1 42 H 31.1.97 20.1.98 19.1.99 23/1 41 H 4.2.97 20.1.98 19.1.99 1.2.00 26/1 5 H 4.2.97 28.1.98 27.1.99 30/1 38 H 4.2.98 28.1.99 31/1 48 H 28.1.98 27.1.99 15.2.00 2/2 112 H 28.1.99 9.2.00 3/2 6 H 18.2.97 31.1.98 27.1.99 11.2.00 6/2 39 H 11.2.97 3.2.98 7/2 66 H 3.2.99 11.2.00 7/2 35 H 26.2.97 3.2.98 27.1.99 8/2 110 H 9.2.00 9/2 115 H 9.2.00 9/2 36 H 26.2.97 10.2.98 28.1.99 11/2 37 H 5.3.97 3.2.98 27.1.99 11/2 114 H 11.2.00 11/2 161 H 11.2.00 11/2

75

Geographic Year 1 Year 2 Year 3 Year 4 Average Variety Region 96/97 97/98 98/99 99/00 Harvest Date 511 KI 21.12.99 21/12 518 KI 21.12.99 21/12 516 KI 29.12.98 29/12 44 KI 22.1.97 20.12.98 13.1.99 8/1 46 KI 11.1.98 13.1.99 12/1 45 KI 22.1.97 15.1.98 12.1.99 16/1 534 KI 19.1.99 19/1 530 KI 28.1.99 28/1 62 KI 26.2.97 3.2.99 11.2.00 13/2 17 MR 17.12.96 4.12.97 19.11.98 9.12.99 4/12 224 N 27.10.99 27/10 230 N 27.10.99 27/10 225 N 1.11.99 1/11 231 N 1.11.99 1/11 61 N 12.11.96 3.11.98 1.11.99 5/11 60 N 20.11.96 17.11.98 18.11.99 18/11 222 N 18.11.99 18/11 10 N 3.12.96 28.11.97 17.11.98 18.11.99 24/11 21 N 25.11.99 25/11 223 N 25.11.99 25/11 280 N 25.11.99 25/11 3A N 1.12.97 24.11.98 27/11 31 N 3.12.97 24.11.98 28/11 63 N 3.12.96 3.12.98 18.11.99 28/11 29 N 9.12.96 1.12.97 19.11.98 29/11 34 N 3.12.96 4.12.97 24.11.98 25.11.99 29/11 25 N 17.12.96 27.11.97 17.11.98 30/11 22 N 19.12.96 3.12.97 19.11.98 25.11.99 1/12 23 N 17.12.96 4.12.97 26.11.98 18.11.99 2/12 232 N 3.12.98 2/12 8 N 19.12.96 3.12.97 19.11.98 3/12 26 N 17.12.96 4.12.97 26.11.98 5/12 30 N 17.12.96 4.12.97 24.11.98 5/12 3B N 11.12.97 1.12.98 6/12 11 N 17.12.97 30.11.99 8/12 24 N 23.12.96 11.12.97 1.12.98 30.11.99 8/12 65 N 8.12.98 8/12 9 N 3.12.97 19.1198 26/12 16 N 22.1.97 15.1.98 13.1.99 16/1 43 SE 12.11.96 18.11.97 3.11.98 11/11

76 6. References

Gail E. Barth and Shona Chinnock, 1999a. Comparisons of yield, quality and floral characteristics of selected and improved cultivars of the Australian wildflower, Ixodia achillaeoides. International Symposium on New Floriculture Crop Development, Chania, Crete, Greece Acta Horticulturae ( in press).

Barth, Gail E, and Shona Chinnock, 1999b. Improvement of Ixodia Daisy for Cut Flower Production. Proc. 5th Australian. Wildflower Conf., Melbourne p 27-28.

Barth, Gail E. 1998. Ixodia Daisy. Chpt in The New Rural Industries: A handbook for farmers and investors. RIRDC, PO Box 4776, Kingston ACT 2604 p.521-526.

Barth, Gail E. 1998. Development of Ixodia achillaeoides as a cut flower crop. Proc. 3rd Int. Symp. on Dev. of New Floriculture Crops, Perth W.A.. Acta Horticulturae 454: 177- 182.

Bennell, M. and G. Barth. 1990. Ixodia as a new cut flower crop. Fact sheet FS/90, Agdex 282/11, South Australian Department of Agriculture:Adelaide.

Copley, P.B. 1982. A taxonomic revision of the Genus Ixodia (Asteraceae). J. Adelaide Bot. Gard. 6(1): 41-54.

Hall, B.H., M.K. Jones, T.J. Wicks, G. Walker and G. Barth. 1996. First report of the diseases of Ixodia achillaeoides in South Australia. Australasian Plant Pathology 25:215.

Maier, N.A., G. Barth and M. Bennell. 1994. Effect of nitrogen, potassium and phosphorous on the yield, growth and nutrient status of Ixodia daisy (Ixodia achillaeoides ssp. alata). Australian J. of Experimental Agriculture 34:681-9.

Rural Industries Research and Development Corporation. 1994. The Australian Flower Industry: A Review. RIRDC Res. Pap. No. 94/9 301p.

Salmon, Alexander. 1996. Breeding approaches to the development of selected native daisies for pot culture . Proc Int. Soc. Plant Prop. 46 (in press).

Weiss, D. and O. Ohana. 1996. Flowering control of Ixodia achillaeoides. Scientia Horticulturae 65 (1) 59-64.

(bold denotes articles produced as a result of this research program)

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