Cutflower Production of Blue Smokebush

A final report prepared for the Rural Industries R & D Corporation by Dr K.A. Seaton

November 2001

ISBN 0 642 58377 3 ISSN 1440-6845

Cutflower Production of Blue Smokebush Publication No. 01/158 Project No. DAW-89A

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 Dr Kevin Seaton Agriculture Western Australia Locked Bag No 4 BENTLEY DELIVERY CENTRE WA 6983 Phone: (08) 9368 3244 Fax: (08) 9367 2625

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]. Website:http://www.rirdc.gov.au

Published in November 2001 Printed on environmentally friendly paper by Canprint Foreword

There is an urgent need to bring new wildflowers into production to diversify production and to command a premium price on export markets. The blue smokebush species, which flower from winter to spring, have particular appeal and bush picked material receives a good price.

The project has characterised a number of blue smokebush selections exhibiting different flowering times, form and colour. Production protocols have been developed for the successful cultivation of blue smokebush. Testing on export markets has shown blue smokebush to be a sought-after cut flower, having specialised use in the ikebana and the wedding trade. With careful marketing this flower should return good profit margins.

This project was funded from RIRDC Core Funds which are provided by the Federal Government

This report, an addition to RIRDC’s diverse range of over 700 research publications, forms part of our Wildflower 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

ii Acknowledgments

The support, cooperation and assistance of Mr Tony Dick and Mr Russell Klopper of Total Flower Exports Pty Ltd are gratefully acknowledged. The support of Mr Digby Growns (project manager Agriculture Western Australia), Mr Mark Webb Kings Gardens and Botanic Park, research colleague Mr Eric Bunn and Mrs Eleanor Bennett formally of KPBG. The assistance of Mr Chris Newell (Tissue culture laboratory manager South Perth) and technical assistants Ms Anouska Cousin, Ms Michelle Crow and Ms Jan Hooper is acknowledged. The work on the project of Medina Research Station manager Mr John Ferguson and the technical assistance of Mr J Berston and Mr Chris McMullan (South Perth) is greatly appreciated. Also the support of Mr Peter Wood (plant pathology AGWEST) and Professor J. Considine (University of Western Australia) is acknowledged as well as the editorial comments of Mr Malcolm Howes (Agriculture Western Australia).

iii Contents

Foreword...... ii Acknowledgments...... iii List of Tables ...... v List of Figures...... vi Executive Summary...... vii 1. Introduction...... 1 2. Objectives ...... 3 3. Methodology...... 4 4. Results:...... 5 4.1. Selection...... 5 4.2. Propagation ...... 6 4.2.1 Initiation of species into culture...... 6 4.2.2 Root initiation and survival...... 8 4.3. Field evaluation...... 11 4.3.1 Establishment and survival ...... 11 4.3.2 Flowering behaviour ...... 11 4.3.3 Stem production...... 13 4.4. Pest and disease control ...... 14 4.5. Postharvest ...... 14 4.6. Production protocol...... 15 5. Discussion of results ...... 16 6. Implications ...... 18 7. Recommendations...... 19 8. Bibliography ...... 20 Appendix I - Agriculture Western Australia Farmnote No. 110/99 ...... 22 Smokebush (Conospermum spp.) for cut flower production...... 22 Appendix II – Photographs ...... 27

iv List of Tables

Table 1. Flower development stages used for assessment flowering of 5 Conospermum spp.

Table 2. Effect of treatment on germination of C. amoenum seeds. The 8 presence of cotyledons are expressed as a percentage of starting number of seeds, and embryos with roots are expressed as a percentage of embryos that produced cotyledons.

Table 3. Effect of hormone type and/or hormone combination on the strike 8 rate of C. brownii.

Table 4. Comparison of selections of blue smokebush (C. eatoniae (Ce), 13 C. caeruleum (Cc) and C. amoenum (Ca)) in terms of flower colour, flowering season and the period flowers were open (days).

Table 5. Stem numbers, stem length and yield of blue smokebush (C. 13 eatoniae (Ce), C. caeruleum (Cc) and C. amoenum (Ca)) selections grown at Medina Research Station.

Table 6. Disease incidence (stems infected per plant) following infection 14 of blue smokebush C. eatoniae and C. caeruleum grown in pots with Phytophthora cinnamomi.

Table 7. Vase life (days) of C. eatoniae (Ce 191 and 196), C. caeruleum 15 (Cc 82) and C. amoenum (Ca 200).

v List of Figures

Figure 1. Flowering score of plants sampled within a natural population 5 of C. eatoniae, showing early and late flowering selections.

Figure 2. Survival of C. amoenum cuttings following initiation into in vitro 7 culture for selection 200, a dark blue selection, pink selection and a juvenile plant.

Figure 3. Root strike rate and plant survival 6 weeks after deflasking from 9 tissue culture for C. eatoniae selections Ce 74, Ce 80, Ce 21A, Ce 24, Ce P5-1, Ce 191, Ce 196 and C. caeruleum Cc 82.

Figure 4. Effect of different rooting treatments on root strike of C. 10 amoenum.

Figure 5. Effect of time of leaving shoots on rooting media on root 10 production expressed as a percentage of shoots tested.

Figure 6. Flowering characteristics of blue smokebush (C. eatoniae (Ce), 12 C. caeruleum (Cc) and C. amoenum (Ca) selections grown at Medina Research Station.

vi Executive Summary

Introduction of novel wildflowers into cut flower production offers considerable marketing advantage. Blue smokebush is one such wildflower, which flowers from winter to spring. The aim of this project was to bring blue smokebush into cultivation. This has involved the selection of germ plasm of three blue smokebush species and the development of a reliable means to propagate these species. It was found that each species had different propagation requirements. For some, clonal propagation was possible using cutting techniques while for others, in vitro tissue culture techniques were necessary. The research has also identified the conditions necessary to maximise shoot and root production in blue smokebush species.

The project has characterised a number of blue smokebush selections exhibiting different flowering times, form and colour. With a blue/dark blue early season (flowering early July-mid Sept), a dark blue mid season (mid July–mid Sept) and a light blue/blue late season (late July -early Oct).

Production protocols have been developed for the successful cultivation of blue smokebush. This package is available as a Farmnote and includes: species selection for a particular climatic zone, site selection, planting layout for maximising yield per row of planting bed, an irrigation and nutritional program, together with pruning, trellising, postharvest treatment and handling strategies.

Testing on export markets has shown blue smokebush to be a sought after cut flower, having specialised use in the ikebana and the wedding trade. With careful marketing this flower should return good profit margins.

vii 1. Introduction

There is an urgent need to bring new wildflowers into production to diversify production and to command a premium price on export markets. A previous RIRDC project, looking at several new wildflowers, identified smokebush (Conospermum spp.) as having cut flower potential (Seaton 1995, 1996, Seaton and Webb 1996). Conospermum spp. occurs in a range of colours including grey/white, cream, pink and blue. (Sainbury 1991, Bennett 1996,) The blue Conospermum species have particular appeal and bush picked material receives a good price on export markets. Blue smoke bush was well received at a FECA ‘Flowerful Australia’ trade exhibition in Tokyo in 1997 (James 1997). Preliminary surveys have identified the blue species C. eatoniae, C. caeruleum and C. amoenum as potential new cut flowers. Presently blue smokebush is almost entirely bush picked and of limited supply with some species not available.

C. eatoniae flowers have good vase life (10 days plus) and have been identified as a potential new cut flower by Japanese importers. It has use in the high value wedding bouquet and ikebana trades. There are also opportunities for blue smokebush to be grown at high density and to gain a share the valuable statice/gypsophila filler flower market as these plants naturally flower in winter when statice and gypsophila require heating and supplementary lighting to flower. Blue smokebush species are generally high yielding and have a long flowering period from July to November. Plantations of blue smokebush (C. eatoniae and C. caeruleum) in Australia have limited plant numbers (approximately between 50 –300 plants). Supplies of nursery stock for planting are also limited because of difficulties in propagation. Blue smokebush plants presently available are limited to one selection each of C.eatoniae and C. caeruleum. C. caeruleum is bush picked from the cooler areas in the south-west of Western Australia. Availability of flowers appear to be uncertain in the future as many populations occur in old growth forest areas in the south-west of Western Australia where picking may be restricted.

C. caeruleum packs well in flower cartons and has favourable freight rates returns per stem on shipments overseas. C. amoenum is quite spectacular in its appearance and has potential to be developed as an excellent export cut flower, having good postharvest handling characteristics. Variation in form and flowering appear to occur in natural populations within these three species of smokebush and this presents an opportunity to develop superior forms on which to build a blue smokebush cut flower industry.

Research is therefore needed to develop these species at a commercial level. Conospermum species are inherently difficult to propagate (Seaton and Webb 1998) and the development of suitable methods of propagation is seen as critical to their production, whether through cuttings, tissue culture or from seed. By working with resources at Agriculture Western Australia, Kings Park Botanic Park and the University of Western Australia, commercial methods of propagation, particularly tissue culture methods, for these species should be possible. The previous RIRDC “New Crops Project” has demonstrated that C. eatoniae can produce commercial quantities of stems (Seaton and Webb 1996), however this is not necessarily the case across the board on flower farms. It is therefore necessary to develop a production

1 system for general application for use by growers such that commercial production (throughout various locations in Australia) is possible.

The project will work closely with industry addressing these problems and informing them of research progress.

In terms of the conservation value of this project, natural populations of these flowers are threatened with extinction from development and are in urgent need of preservation. Many of the natural stands of C. eatoniae have been lost through development and farming. Although C. amoenum was once common in the Perth region (Bennett 1996) surveys have shown that there is now only two known stands, and these are being lost to urbanisation and mining. One effective way to preserve these species is through commercially growing selections for cut flower production. By surveying and collecting a range of types from natural populations and propagating these plants this project will greatly contribute to the species preservation.

2 2. Objectives

To provide the wildflower industry with new opportunities to capture a share of the highly profitable cutflower markets, previously reserved for exotic flowers, by developing blue smokebush (Conospermum spp.), to be sold as a feature filler over an extended season. The blue smokebush species being targeted in this project are: Conospermum eatoniae, Conospermum caeruleum, Conospermum amoenum

3 3. Methodology

Selection. - To define natural populations of C. eatoniae, C. caeruleum and C. amoenum, selecting suitable mother plants in terms of colour, ease of propagation, vigour, stem length and display and flowering time.

Take steps necessary to preserve natural populations of these species in terms of priority listing, maintaining mother plants in stock gardens.

Propagation. - Develop reliable means of propagation to commercial levels. This will involve investigating cutting (propagule, hormone, time to propagate), tissue culture and seed germination as means of propagating plants.

Evaluation. - Establish field plantings of different selections and determine response in terms of production, stem length and flowering. Identify new selections suitable for commercial cut flower production.

Postharvest. - Determine the postharvest vase life of selections and postharvest handling characteristics.

Production protocol. - Determine protocols for maximising production in a commercial situation. Determine methods of establishment, optimum densities, water and nutrient management to maximise stem length and quality. Determine suitable pruning methods. Determine correct postharvest handling procedures for fresh and dried product. Conduct test shipment trials through exporters to determine the performance of blue smokebush on export markets.

Commercialisation. -Organise grower groups, produce printed material to provide details on production of these species and hold demonstrations and field days.

Data were analysed using ANOVA and linear regression analysis using the Genstat® IV statistical package. Mean separation was determined by least significant difference at P = 0.05. Means of treatments and standard errors of means are presented where appropriate.

4 4. Results:

4.1. Selection

C. eatoniae. -Plants were surveyed fortnightly during the flowering season and flowering stages scored according to the following scale (Table 1.).

Table 1. Flower development stages used for rating of flowering of Conospermum spp. Flowering stage Description 1 vegetative buds only present 2 tight flowering buds present 3 flower buds swollen, beginning to open 4 10-25% of flowers open on bush 5 25-50% of flowers open on bush 6 50 – 75% of flowers open on bush 7 75 – 100% of flowers fully open 8 flowers fading and or shedding, (seeds may be present)

Plants were identified which ranged in flowering from early to late types (Fig. 1). For the early flowering plants, flowers opened rapidly reaching stage 5 (or 50% open) by mid June while for the late flowering types, flowers opened more slowly reaching stage 5, by late August. Selections were made of early, late and intermediate flowering types. 9 8 7 6 5 early 4 late 3 Flowering score 2 1 0 0 0 0 000 000 2000 2000 2000 8/20009/2000 05/ 05/ 05/ 0 0 03/ 17/ 31/ 14/06/20028/06/20012/07/20026/07/209/08/223/ 06/ 20/09/2000

Figure 1. Flowering score of plants sampled within a natural population of C. eatoniae, showing early and late flowering selections.

Selections were made with flower perianth colours of light blue (RHS 95A), blue (96A), and dark blue (100A).

5 Plant growth habit ranged from compact with high stem density to tall with sparser stem density. Stem length ranged from 400 mm to 900 mm and stem numbers ranged from 15 to 90 stems per bush.

C. caeruleum. -Populations were surveyed and two forms were identified the eastern more sprawling forms reaching up to 2 m in diameter and the forest forms with a more upright habit.

C. amoenum. - Populations were surveyed and plants identified exhibiting perianth colours of light blue (RHS 92C), blue (92B), deep blue (92B), purple (91C), red (55C) and white (91D). In one population, light blue, blue and deep blue plants and purple occurred in the proportions of 42.4, 31.8, 13.6 and 12.1% respectively. A few white plants and one red plant occurred in other populations. Cuttings and seeds were collected for propagation. Plant selected varied in flowering from mid August to mid October. Stem length ranged from 48 to 75 cm.

4.2. Propagation

4.2.1 Initiation of species into culture

Seeds and cuttings. -Germination of mature seed in seed raising mixes with or with out smoke treatment collected from these species was found to be unreliable with <1% of the seeds germinating. Tests showed that C. eatoniae and C. caeruleum did not strike roots from cuttings using a range of cutting material including basal buds. Strike rate of cuttings of C. amoenum collected from plants in natural populations was very poor. Out of 18 selections only 2 propagated and these had a strike rate of 8 and 24%. These plants were established as mother plants at Medina Research Centre. Cuttings taken in late spring/summer from cultivated plants struck roots in 3% IBA after nearly 3 months under fog on heat beds (26 °C), and had a strike rate of 57.2 ± 2.7 %.

Tissue culture. -Tissue culture methods were developed suitable for these Conospermum species. The involved either embryo rescue of seed collected or initiation of cuttings into in vitro culture.

C.eatoniae and C. caeruleum were most successfully initiated into culture using newly elongated vegetative shoots, which developed after flowering. Fifteen lines were initiated into culture and 7 lines were identified as performing well in culture.

C. amoenum.-It proved more difficult to initiate clonal material of C. amoenum selections into culture than for C. eatoniae and C. caeruleum. The survival of some of the lines of C. amoenum following initiation is shown (Fig. 2). Two lines of C. amoenum were successfully initiated into culture out of the 13 tested. Cuttings were initiated into ½ MS (Murashige and Skoog 1962, Taji, 1995) tissue culture media using 80 mL tubes supplemented with, 60 mM sucrose, pH adjusted to 6.9 (before autoclave), 8 g/L agar and 6-benzylaminopurine (BAP) plus indole-3-butyric acid (IBA). The addition of cytokinins (kinetin or BAP) was found to encourage shoot proliferation (Bunn 1994) and auxin (IBA) encouraged shoot development (Newell pers. comm.). A two-stage disinfestation process for cleaning field material before

6 initiation was used. This involved a 5 sec dip in 0.1% sodium hypochlorite, draining and wrapping in moist paper towel and storing in plastic bags for 24 hours. This was followed by a second wash in 1% sodium hypochlorite for 1 minute with 0.05% Tween80® detergent added and brushing to ensure all parts of the material were washed. The shoot material was then thoroughly washed in sterile water and initiated under sterile conditions into tissue culture. Cultures were maintained at 25°C, 16/8 hour light/dark cycle under fluorescent light with a photo flux density of 40 µM.m- 2.sec-1.

Loss of cultures during initiation was due to contamination as well as attrition (either due to damage from the disinfestation process or senescence of the cutting). Approximately 75 days after initiation there was sufficient multiplication of selection 200 for numbers to increase such that by day 175 or 6 months after initiation there was sufficient material for root initiation trials (Fig. 2).

250

200 sel. 200 150 d. blue pink 100 Juvenile 50 survival/multiplication (%) 0 0 50 100 150 200 time (days)

Figure 2. Survival and multiplication of C. amoenum cuttings following initiation into in vitro culture for selection 200, a dark blue selection, pink selection and a juvenile plant.

Embryo rescue. -None of the seed of 3 selections of C. amoenum, with or without a small portion of testa removed, germinated after 4 weeks at 20°C (Table 2). Viability tests on seed showed that 100% of seeds contained endosperm. By using embryo rescue techniques, to remove and grow the embryo with endosperm in vitro, between 80 and 100% of embryos survived and produced cotyledons and of those 28 to 100% produced a root radicle after 4 weeks.

7 Table 2. Effect of treatment on germination of C. amoenum seeds. The presence of cotyledons is expressed as a percentage of starting number of seeds, and embryos that developed roots are expressed as a percentage of embryos that produced cotyledons.

Means followed by the same letter are not significantly different at P = 0.05.

Selection Endosperm Germination Embryo rescue (%) present (%) Seed coat Seed coat Cotyledons Roots intact damaged 1 (96/11) blue 100 0 0 100 a 100 ± 15.3ab 2 (96/2) blue 100 0 0 80 ± 13.3a 28.6 ± 17.2 b 3 (96/1) blue 100 0 0 80 ± 13.3a 42.9 ± 18.9 b

Therefore embryo rescue appears to overcome the problems of seed coat induced dormancy encountered in germinating hard seeded plants. The technique proved to be a much more efficient means of initiating selections of C. amoenum into culture than using clonal material. The disadvantage was that plants initiated were not clonal.

4.2.2 Root initiation and survival

A) C. eatoniae and C. caeruleum A range of hormones were tested on blue Conospermum species, initially on a C. brownii as part of a student honours project supervised during this RIRDC project (Cousin 1998). Auxins tested included indole-3-butyric acid (IBA), napthalene acetic acid (NAA), indole-3-acetic acid (IAA) and 2,4-dichlorophenoxyacetic acid (2,4-D). These were tested as a single auxin or in combination and at concentrations ranging from 1.25 µM (micro moles/litre) to 10 µM. NAA and IBA + NAA gave significantly higher root strike than other treatments (Table 3).

Table 3. Effect of hormone type and/or hormone combination on the strike rate of C. brownii. Means followed by the same letter are not significantly different at P = 0.05.

Treatment Hormone Root strike rate (%) 1 IAA 0 c 2 IBA 33 b 3 NAA 100 a 4 2,4-D 0 c 5 IAA + IBA 47 b 6 IBA + NAA 100 a

Applying the best two hormones treatments 3 and 6 (from C. brownii, Table 3) to C. eatoniae and C. caeruleum gave a 3 fold increase in root strike compared to other treatments. These hormone treatments were then used for screening other selections of C. eatoniae for rooting ability.

Root strike of C. caeruleum selection Cc 82 and C. eatoniae selections Ce P5-1 and Ce 80 was 63% to 73%, which was significantly higher than that for Ce 74, Ce 24, Ce

8 191 and Ce 196 which was 20 to 26%. The strike rate of Ce 21A was the lowest at 6% (Fig 4).

100

60 root strike Plant survival 40

20 Rootstrike rate and survival (%)

0 74 80 21A 24 P5-1 191 196 82 selection

Figure 3 Root strike rate and survival 6 weeks after deflasking from tissue culture for C. eatoniae selections Ce 74, Ce 80, Ce 21A, Ce 24, Ce P5-1, Ce 191, Ce 196 and C. caeruleum Cc 82. Data show means ± standard error of means, (s.e.).

Survival on deflasking from tissue culture of selections Ce 80, Ce P5-1,Ce 196, Ce 24 and Cc 82 was significantly higher survival (57 % to 73%) than Ce 74, Ce 191 and Ce 21A. (25 to 36%) (Fig. 3). Higher root strike rate was reflected in higher survival rates except for Ce 21A and Ce 24 (Fig. 3). After 6 weeks plants were ready for planting in the field, holding plants in a glasshouse for a longer periods resulted in further losses.

B) C. amoenum Once established in culture C. amoenum shoots were easier to multiply compared with C.eatoniae. However, in contrast it was found to be more difficult to initiate roots in C. amoenum than C.eatoniae. Results in Figure 5 show that treatments 3, 4 and 5 gave 75 to 100% root strike. Treatment 2 gave 10% root strike and the control (treatment No. 1) with no added hormone initiated no roots(Fig 4).

9 100

40 Root strike rate (%)

0 012345 Rooting treatment

Figure 4. Effect of different rooting treatments on root strike of C. amoenum. Data show means ± standard error of means, (s.e.).

Also it was found the time shoots remained on rooting media was important for improving root strike with a critical time of between 7 and 14 days (Fig 5). By leaving shoots on the media too short or too long reduced the root strike percentage. Longer times than 14 days on rooting media favoured the production of masses of fine roots instead of fewer, thicker, more robust roots,

100

Root strike rate (%) 20

0 5 10 15 20 Time on media (days)

Figure 5. Effect of time of leaving shoots on rooting media on root strike expressed as a percentage of shoots tested. Data show means ± standard error of means, (s.e.).

10 Survival of shoots from tissue culture was 67%, however, plants were susceptible to disease and regular fungicide control for Botrytis cinerea and Pythium sp.was imperative to achieve this level of plant survival.

4.3. Field evaluation

Selections were grown at Medina Research Station and evaluated for flowering time, form, production and postharvest.

4.3.1 Establishment and survival

Blue smokebush plants C. eatoniae (Ce) and C caeruleum (Cc) established well at Medina Research Centre. Three months after a November planting, an average of 90 ± 5% of plants remained in the field. After 6 months, 89 ± 5% of plants were alive and after 12 months 71 ± 9% of plants remained. Survival of selections Ce P5-1, Ce 74, Ce 80 was highest with 87.5-100% of plants remaining, while Ce 24 was lowest with only 22% remaining after 12 months. Survival of the selections Ce 74, Ce 191,Ce 196 and Cc82 12 months after planting was between 60-75%. For the C. amoenum selection, Ca 200, which was planted at a different time from the other blue smokebush selections, survival was 56 ± 15% 12 months after planting.

4.3.2 Flowering behaviour

There was a significant difference in terms of flower development for the nine blue smokebush selections tested in the field at Medina (Fig. 6). The spread of flower development times between selections was noticeable when compared at the 50% flowers open (stage 5) (Fig. 6). The earliest selections (Ce 21A, 191 and 80) commencing flowering on 9-12 July and reached 50% on 2 August and were 100% open on 15 t - 17 September. Whereas the longer flowering selections Ce 74 and Ce 24 flowered for another 15 days than these earlier selections. The later selection P5-1 commenced flowering on the 28 of July, 12 days later than Ce 196 (16 July) and reached 100% open 9 days later than Ce 196 on the 6 October. Selection Ce 74 and Ce 24 were relative slow to open taking 80 plus days compared with other selections taking 64 to 70 days (Table 4.) C. amoenum Ca 200 was later by 12 days to commence flowering compared with Ce P5-1, although this selection reached 50% flowering 7 days sooner than Ce P5-1 and flower opening was shorter, taking 56 days.

11 Ce 21A 50% 100% Ce 191 l.s.d P=0.05

Ce 80

Ce 74

Ce 24

Ce 196

Cc 82

CeP5-1

Ca 200

182 192 202 212 222 232 242 252 262 272 282 292

July August September October

Figure 6. Flowering characteristics of blue smokebush (C. eatoniae (Ce), C. caeruleum (Cc) and C. amoenum (Ca) selections grown at Medina Research Station (C. amoenum was planted in a different year and was not compared in the statistical analysis of the other selections). Flowering times include; from first flower opening, 50% open and 100% open. Least significant difference (l.s.d.) at (P = 0.05) is shown.

Form these results several selections of blue smokebush exhibit potential to be marketed as new cut flowers. Selections identified have a range of colour, flowering times and flower duration (Table 4). With a dark blue early season (flowering early July-mid Sept), a light and dark blue mid season (mid July–late Sept) a light blue late season (late July -early Oct) and a light blue very late season (mid Sept to mid Oct) selection.

12 Table 4. Comparison of selections of blue smokebush (C. eatoniae (Ce), C. caeruleum (Cc) and C. amoenum (Ca)) in terms of flower colour, flowering season and the period flower were open (measured from opening (stage 3, Table 1, to 100 % open (stage 7)) (days).

Means followed by the same letter are not significantly different at P = 0.05.

Species selection Flower colour (RHS) Flowering Duration of season opening (days) C. eatoniae Ce21 Dark blue (94A) Early 68 b Ce 191 Blue (96A) early 70 a Ce 80 Dark blue (100A) early 65 b Ce 74 Early (light blue (95A) early 80 a Ce 24 Dark blue (100A) Early-mid 81 a Ce 196 Dark blue (96A) Early 66 b Ce P5-1 Light blue (95A) late 66 b C. caeruleum Cc 82 Dark blue (96A) mid-late 64 b C. amoenum Ca 200 Blue (96A) late 56* * not included in statistical analysis

4.3.3 Stem production

Selections varied in terms if stem length and stem production (Table 5). Ce 80 produced the most stems, however, stem length was short at 41 cm in the first year While in contrast Ce P5-1 stem length was 67 cm with stem production 147 stems/m2.

Table 5. Stem numbers, stem length and yield of blue smokebush (C. eatoniae (Ce), C. caeruleum (Cc) and C. amoenum (Ca)) selections grown at Medina Research Station. Data are means, means in columns followed by the same letter are not significantly different at P = 0.05

Selection Stem No. Stem Yield (Stems/ length (stems/ m2) plant) (cm) Ce 80 111 41.4 222 a Ce 74 35 50.8 74.4 c Ce 24 30 38.5 60 d Ce 21A 34 44.2 68 c Ce P5-1 74 67.2 147.5 b Ce 191 22.6 81.2 47.4 d Ce 196 26.3 72.2 52.6 Cc 82 37 82.7 73.5 c Ca 200 48.7 55.0 97.4* * C. amoenum was not included in the statistical analysis

13 Plantings of C. amoenum Medina Research Station were made at a different time than for C. eatoniae and C. caeruleum selections and results were analysed separately. Stem yield per plant for 18 month old plants was 48.7 ± 3.7, with a stem length of 50- 60 cm. Plant height was 61 ± 9 cm and width of 97 ± 18 cm. Flowering stems were tight racemes with flowers along 50% of the stem giving a good flower display.

4.4. Pest and disease control

Pot tests showed that both C. eatoniae and C. caeruleum were susceptible to infection by Phytophthora cinnamoii (Table 6). The site should also be free from Phytophthora spp.i.e. dieback Indicating the necessity to select and protect the site from infection by die back. This involves ensuring strict quarantine cleaning treatments such as foot baths and water chlorination or UV treatment. Also only buying stock plants from an accredited nurseries to prevent entry of Phytophthora spp. onto the site.

Table 6. Disease incidence (stems infected per plant) following infection of blue smokebush C. eatoniae and C. caeruleum grown in pots with Phytophthora cinnamomi Agriculture Western Australia isolate No. 2864. Plants were scored for disease severity using the following scale: 0, healthy and no wilting, 1, 20% 2, 40%, 3, 60%, 4, 80% and 5, 100% wilting. Data are means, values in columns followed by the same letter are not significantly different at P = 0.05

Species Disease incidence Disease score (%) Control 0 b 0 b C. eatoniae 100 a 3.93 a C. caeruleum 93.3 a 4.20 a

Other fungal diseases such as Botrytis spp. and Alternaria spp. do not appear to infect blue smoke bush species at Medina. The plant does not require a regular spray program for pests or disease. In cooler conditions the situation may be different although there is no evidence of this at sites growing C. caeruleum at Albany 350 km south-east of Perth.

4.5. Postharvest

Vase life varied from 9 to 14 days for different selections (Table 7.) for flowers standing in vases of distilled water. No special vase solutions were necessary for blue smokebush.

Vase life of flowers was measured only for C. eatoniae and C. caeruleum as these species had no leaves on the flowering stems. Other selections of C. eatoniae were not rigorously tested although observation of flowers picked for vases showed these all had a vase life within the 10 to 14 day period. Leaves were tested for vase life as well as flowers in C. amoenum. Stems of C. eatoniae are packed as 5-10 stem bunches, sleeved (perforated) with 30 bunches per 12 kg carton. More bunches of C. caeruleum can be packed per carton without the need for sleeving. C. amoenum packs in 5 stem bunches with 20 bunches per carton.

14 Table 7. Vase life (days) of C. eatoniae (Ce 191 and 196), C. caeruleum (Cc 82) and C. amoenum (Ca 200).

Data show means ± standard error of means, (s.e.).

Species Vase life (days) Flower Leaf C. eatoniae Ce 191 10.8 ±0.8 - Ce 196 8.8 ± 1.1 - C. caeruleum 13.5 ± 1.2 - C. amoenum 13.8 ± 1.9 14.9 ± 1.0

4.6. Production protocol

The information necessary for successfully producing blue smoke bush as a commercial cut flower has been detailed in the following farmnote. Cultivation of smokebush (Conospermum spp.) (Appendix I). This Farmnote has been made available to the industry. It contains information on blue smokebush together with white smokebush (another group of smokebush which do not from part of this project but are also being developed by Agriculture WA as a cut flower and which compliments blue smokebush production). The Farmnote provide information on;

(a) species selection (b) site selection (c) method of establishment (d) nutrition and irrigation (e) pest and disease control (f) pruning and trellising (g) postharvest management

15 5. Discussion of results

The scope of this project was broad in the sense of identifying potential new wildflowers from wild populations, collecting these and propagating plants, then cultivating plants and testing their response to cultivation. The requirement of the project was to complete this process by providing the wildflower industry with new cut flowers suitable for commercial production together with a production protocol for successful stem production. To achieve this objective a number of hurdles have been overcome, new techniques have been developed and our knowledge of the requirements needed to develop new wildflowers greatly increased. The surveys done in the project showed there was the loss of many natural populations of blue smokebush especially C. eatoniae and C. amoenum through clearing or urbanisation. The genetic material collected in the project therefore is a valuable source of material for conservation of these species. Surveys showed there to be a large variation in form, colour and flowering time of blue smokebush species. This opens up a much greater market window adding variety to the blue smokebush that are available. By having flowers available from July to late October, as is possible using species selected in this project, increases the value of this crop to the Australian cut flower industry.

Conventional methods of using cuttings and germinating seeds were found to be inadequate for C.eatoniae and C. caeruleum and the project concentrated on the use of tissue culture methods for these species. It is now possible to reliably propagate blue smokebush using tissue culture methods developed in the project. These methods have been tested to produce plants with well-developed root systems to provide plants suitable for establishment in the field. For C. eatoniae a long lead time from collection of new wild material to establishment into culture was required. This could be up 18 months to two years. It was possible to achieve a multiplication rate of 2-3 times in culture provided suitable procedures were followed.

Success in root initiation was found to vary with the media and auxin used. The project has established a hormone treatment and concentration that reliably produced roots in in vitro culture. The response to this hormone treatment varied with the particular selection and this also depended on the time the plant had been in culture.

The blue smokebush species C. amoenum was found much more difficult to propagate from cuttings using wild material. However, using material from cultivated plants, techniques were developed (which included frequent pruning back of parent material) and gave 57% strike rates provided cuttings were taken at a particular time of year. This compares favourably with 0 to 24% for cuttings taken from wild material. Tissue culture methods using embryo rescue techniques were effective in initiating selections of C. amoenum whereas clonal material was much more difficult. Methods were also developed to achieve good multiplication rates and root initiation. Further work is needed to reliably deflask rooted plants from culture.

Management practices are now available in the form of a Farmnote. Information is also given for these smokebush species in terms of site selection and planting bed preparation, fertiliser and irrigation requirements, as well as pest and disease management. The Farmnote pays special attention to the special requirements of blue

16 smokebush species in terms of site selection, dieback sensitivity, specialised nutrition requirements and irrigation scheduling, trellising, pruning as well as postharvest handling, transport and market uses.

The flowers selected appear free from damaging insects and fungal infection, with little insect loads and mild postharvest treatment; export systems should be more than adequate to cope with these. They have adequate postharvest life although care is needed with postharvest handling to ensure that plant stems are kept hydrated during handling. Sleeving can be used to prevent crushing and tangling when packing for export. Plants would also benefit from maintaining a cool chain.

Test marketing has indicated that demand for blue smokebush is strong with good prices. Having a product available for a longer period as these new selections provide would maintain the consistency of supply and benefit sales. Blue smokebush also offers a unique Australian native wildflower alternative to gypsophila.

17 6. Implications

Introduction of new and interesting wildflowers into export markets is seen as essential to maintaining Australia’s market share. Test marketing during the project has indicated that blue smokebush is a sought after wildflower with use in the upper end of the cut flower trade. It is bought in Japan particularly for special use in ikebana and the wedding bouquet industry as well as providing backdrop to focal displays at conventions and in trade promotions where the blue smokebush is used as a feature filler with its eye catching blue colour and fine flower detail. It is a very adaptable cut flower and can be used en masse to offset a background to central feature flowers such as kangaroo paw, proteas or exotics such as tulips. It can also create interest as single stems in a flower display.

Selections of blue smokebush have been identified from bush populations that exhibit a range of colours from light blue to dark blue and flowering in Perth Western Australia from the first week in July to the first week of October. These selections have been also been characterised from field trialing indicating several with long stem length (70-80 cm) and stem yields of 50-150 stems per square metre. Selections have also been identified which have higher root strike.

We are now aware that the blue smokebush species are not easily propagated from seed, with C.eatoniae and C. caeruleum only being able to be propagated through tissue culture techniques. C. amoenum can be propagated through either tissue culture or cutting propagation techniques although at this stage it is better to propagate C. amoenum using cuttings taken from a cultivated mother plant that has been identified as having a high strike rate.

Information on field management of blue smokebush is now available as a Farnmnote. It provides a protocol for the successful cultivation of blue smokebush and covers aspects of plant characteristics. It should be stressed that blue smoke bush is sensitive to high levels of phosphorous and a fertilser program needs to take this into account. Blue smokebush performs best when irrigated using trickle irrigation. Also plants are susceptible to dieback and that strict site hygiene needs to be practiced. Plants are reasonably tough in terms of postharvest management provided that flowers are kept hydrated.

18 7. Recommendations

Several selections of blue smokebush have been identified which may be developed commercially in the future. They have different forms and a spread of flowering times. Growing some of these would help to open up the marketing window of blue smokebush, and overcome the problems of being restricted to one or two varieties, as is the present situation.

This project has shown that at an experimental level it is possible to propagate blue smokebush given certain protocols are followed. The best chance of successfully achieving this at a commercial level, and make plants available, depends on industry. What is needed is a specialist nursery with the capability to develop the infrastructure and know how to implement propagation systems suitable for the various species of blue smokebush. Their chance of success will increase by using the high strike rate selections of blue smokebush identified in the project.

Field management protocols have been worked out and are available as farmnote. It covers aspects of site preparation, plant establishment, and irrigation methods. Together with fertiliser program and plant trellising, pruning strategies and posthavest handling practices suited to producing blue smoke bush. Growers should adapt these protocols to suit their own farm situation.

19 8. Bibliography

Anon 2000. Annual report Centre for Australian Plants (CAP) 1998-1999. Agriculture Western Australia. 3 Baron Hay Court, South Perth .

Anon 2001. Annual report Centre for Australian Plants (CAP) 1999-2000. Agriculture Western Australia. 3 Baron Hay Court, South Perth .

Bennett, E.M. 1996. Flora of Australia Vol 16. Elaeagnaceae, Proteacea 1. Conospermum. pp 224-486.

Bunn, E. 1994. In vitro propagation of Australian Proteacea. Short report. Kings Park and Botanic Gardens 3pp.

Cousin, A 1998. Changing the state of cell division from shoots to roots in two recalcitrant native woody species: Conospermum brownii and Geleznowia verrucosa, using various auxin treatments in vitro. Honours thesis. Faculty of Agriculture, University of Western Australia. Nov. 1998. pp1-65.

James, K. 1997. Flowerful Australia’ on display in Tokyo. The Floriculture industry Newsletter. No. 43, December. pp 2-3.

Murashige, T. and Skoog, F., 1962 A revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol. Plant. 15: 473-479.

Seaton, K. A. (1995). Introduction of new wildflowers into cultivation. J. West. Aust. Wildflower Producers Assoc. Sept. 1995. p8.

Seaton, K. A. 1996. Conospermum in the search for the new wildflower. Australian. Horticulture. June-July 1996. 4(6): 47-48.

Seaton, K.A. 1999. The introduction of new wildflowers into commercial production. Floriculture Industry Newsletter. No. 47, . p.6.

Seaton, K.A. 2000. Cultivation of smokebush (Conospermum spp.) for cut flower production. Farmnote 110/99. Agriculture Western Australia.

Seaton, K.A. 2000. New developments in blue smokebush research. Floriculture Industry Newsletter. No. 52, December 2000. p.17.

Seaton, K. A. and Webb, M. G. 1996a. Production management and handling of quality new native cutflowers for export. RIRDC final report. 1996. 50 pp.

Seaton, K.A., and Webb, M.G. 1998. Conospermum : a cultivated cutflower. Proc. 26th Annual Conference International Plant Propagators Society Ltd. Perth WA. . 4pp.

20 Sainbury, R.M. 1994. A field guide to smokebush and honeysuckle (Conospermum and Lambertia). University of Western Australia Press. 1994. 97 pp.

Taji, A.M., Dodd, W.A., and Williams, R.P., 1995. Plant Tissue Culture Practice. University of New England, Armidale, NSW. 129 pp.

Tan, B., de. M. Vos and M. Vos. De. 1994. Tree smokebush. Conospermum triplinervium. In vitro germination and propagation. Australian Plants. 18: 28-35.

21 Appendix I - Agriculture Western Australia Farmnote No. 110/99

Smokebush (Conospermum spp.) for cut flower production By Kevin Seaton, Research Officer, Agriculture Western Australia, South Perth Introduction New plants have to be brought into cultivation to sustain the growth and competitiveness of the wildflower industry. Two such plants are Conospermum spp. and Pimelea physodes. Growing new species of export wildflowers may require development of new cultural methods that differ from those used for well established varieties. Returns are generally higher with these new species and trials at Agriculture Western Australia’s field stations have demonstrated that they can be cultivated successfully. Test marketing has also shown that many of these wildflowers can be profitable. One wildflower presently being bush picked is smokebush. Smokebush, is in the family Proteaceae and are members of the genus Conospernum. They offer a wide diversity of flower types ranging from white to blue and flowering from summer to winter. They are almost entirely picked from natural populations and are exported fresh or dried to Japan, Europe and the USA. Species selection Six species of Conospermum have been selected that can be grown as cut flowers. These include both blue flowering types C. eatoniae (blue lace) and C. caeruleum (slender smokebush), and white or grey flowering types C. triplinervium (tree smoke), C. stoechadis (common smokebush), C. incurvum (plume smokebush), and C. crassinervium (tassel smokebush) (Table 1). Conospernum eatoniae has leafless stems with masses of clusters of china blue flowers; it is sought after on Japanese markets. C. caeruleum has large flat leaves that remain on stems with fewer blue flower clusters than C. eatoniae. C. stoechadis has a grey/white flower panicle and narrow leaves and is a high stem producer. C. triplinervium comprises a number of species with broad leaves with various forms from weeping to strong upright stems. One particular form of C. triplinervium has heavy stems and white hairy flowers; it has been named the “elk” smokebush. C. incurvum is a smaller plant than C. triplinervium or stoechadis and has spikes of leafless stem covered in white flowers with a black tip to the perianth. C. crassinervium grows fairly slowly and produces flowers in a corymb forming a tassel. It flowers in summer but continues to flower over an extended period in cultivation.

22 Table 1a. Cut flower characteristics of smokebush species - Blue Flowered Species C. eatoniae C. caeruleum Common name Blue lace slender smokebush Growth habit upright spreading Flowering period July-Aug July-Oct Average production* high more than 50 stems medium 25-50 stems per plant Comments prefers warm climates, in prefers cooler high demand climates, packs well

Table 1b. Cut flower characteristics of smokebush species - White/grey flowered Species C. crassinervium C. triplinervium C. stoechadis C. incurvum Common name tassel smokebush elk smokebush common plume smokebush smokebush Growth habit upright upright spreading to spreading to upright upright Flowering Dec-Feb Aug-Nov July-Oct Aug-Oct period Average low less than 25 high more than high more than medium 25-50 production* stems 50 stems per 50 stems per stems plant plant Comments late flowering, wide area high stem small shrub, dries well suitability range production attractive flowers of types Climate Smokebush can be grown in a wide range of rainfall zones from 350 mm to 850 mm and maximum/minimum average temperature 23/10°C. However, stem production may be lower and the flowering period may be shorter if conditions are too extreme. For example, C. eatoniae performs better in drier, warmer regions while C. caeruleum flowers for a longer period in cooler regions. Smokebush has some frost tolerance. Site requirements Choose a weed free and diseases free site. Preferably one with minimal or no previous fertiliser history. Smokebush prefer free drained, deep acid sand (pH 5.5 to 65 in water). Smokebush is fairly intolerant of residual phosphorus in the soil. Some species will tolerate loamy soils provided they are freely drained. Early weed control is required to significantly reduce weed presence. This can be achieved by cultivation, knockdown herbicides (such as glyphosate and Spray.Seed) and a pre emergence treatment. Planting and establishment Weed control needs to be maintained during establishment to ensure plant survival. Weedmat can be used or alternatively laying 75 to 100 mm of bark mulch aids establishment, suppresses weeds, conserves soil moisture and reduces soil temperature during summer. Also the use of covers such as ‘grow cones’ or similar tree guards aids establishment, particularly on windy sites, and can discourage native animals from nibbling young foliage. Transplants should be greater than l0 cm high. .Planting density varies depending on species from 3300 to 9500 plants/ha. For C. eatoniae,C. caeruleum and C. incurvum plant in double rows, 0.6 m apart and with 0.7 to 1 m between plants on a staggered grid. This gives optimum production. For other species of smokebush plant in single rows with spacing of 1 m between plants in beds 3 m apart.

23 Fertilisers Smokebush have low requirements for nutrients, particularly phosphorus. Our research has shown that they also tolerate low levels of micro nutrients, particularly manganese and zinc. On planting, 10 gram of low phosphorus slow release such as ‘osmocote’ or ‘nutricote’ (low P) should be spread on the soil surface adjacent to plants. Once established, plants after 2 -3 months require a regular feed preferably through the irrigation lines such that plant receive 65-90 kg/ha/annum of nitrogen and 60-90 kg/ha/annum potassium, 8-12 kg/ha/annum of phosphorus, 7-10 kg/ha/annum of calcium together with a balance of trace elements. Injecting fertilisers during watering (fertigation) to meet the requirements of Pimelea physodes can be achieved by making up a stock solution of the following fertilisers in a 200 litre drum of water;

Fertiliser Amount (g/200 litres) Agran 1120 Potassium nitrate 1760 MAP (mono ammonium 400 phosphate) Calcium nitrate 320 trace elements (Librel 32 BMX)

In summer fertigate plants on a daily basis. The frequency of fertiliser injection should be scaled back during autumn to every third day in winter then increasing in frequency through spring. During each watering allow the irrigation system to run for 10 minutes, then inject over the next 40 minutes 10 litres of the stock solution per 800 plants being watered, followed by 10 minutes of watering without fertiliser. Acclimatise young plants to the above fertiliser program by gradually increasing the frequency of injection i.e. first inject fertiliser one day out every three days for a two week period, increasing the frequency until plants are fertigated daily. (refer to Farmnote 99/94 for selection of fertigation equipment). Irrigation Smokebush should be planted into moist soil and watered regularly during establishment using drippers, particularly on sandy soils. In summer watering may need to be split into 3 applications per day. This is best done using an automatic watering system. Once established blue smokebush species are low water users (i.e. 50% evaporation replacement or 4 litres per day per plant in summer). White/grey smoke bush are higher water users (60% evaporation replacement or 5 litres per day in summer).

24 Trellising Conospernum eatoniae need to be supported in their first two years of growth to prevent wind damage, this can be easily achieved by supporting plants on 15 x 15 cm square wire trellis or flower mesh (Cyclone) when 15 to 20 cm high or 2-3 months after planting. White smokebush may need staking in its first year of growth but the stakes should be then removed to strengthen the plant stem. Pruning Smokebush must be pruned in the first year of growth to develop a good productive branch structure in white smokebush or to encourage new stem production in blue smokebush. For white smokebush this is done by cutting back new branches to 10-15 cm from the main stem, leaving green growth below the cut. In the second and subsequent years this process is repeated but on the secondary and tertiary branches. Stem production should increase up to five fold per year. For blue smokebush, all new stems should be pruned off after flowering to 10-20 cm once new stems have appeared taking care to leave new stem growth on the plant. Pest and diseases Very few pests attack smokebush and, except for very late flowering selections, it is rare for flowers to harbor large insect loads that may cause a problem with quarantine . Caterpillars (Lepidoptera) may attack smokebush and this can cause stems to collapse, particularly in C. eatoniae and C. caeruleum. Catasarcus weevil can cause leaf damage on C. triplinervium. These pests can usually be controlled by strategic sprays of permethrin or alpha-cypermethrin. Thrips (Thysanoptera) in flowers may need to be sprayed before picking with fluvalinate or similar insecticides. Conospermum is susceptible to Phytophthora spp. or dieback and Pythium. Test soil and nursery stock before planting to avoid these diseases. Once plants are affected they seldom recover but applications of Fosject may be beneficial. Postharvest treatment and transport Conospermum do not require any special vase solutions but blue species must have their stems stood in water after picking to maximise vase life and quality . C. eatoniae are best picked and packed in 5 stem bunches, in perforated sleeves to prevent tangling . White smokebush respond to 5-10% glycerin for preserving stems. Flowers can be treated for insect disinfestation with an Insectigas/Pestigasaerosol treatment. Bunches should be cooled to 2°C before export. Marketing Blue smokebush is a sought on overseas markets, particularly in Japan as a high value use cut flower for use in the wedding and Ikebana trade. White smokebush is being used as a filler and generally commands a lower price than blue smokebush. Further reading • Farmnote no.111/99 'Qualup bell (Pimelea physodes) for cut flower production' • Farmnote no. 99/94 'Selection of fertigation equipment' • Farmnote no. 1/96 'Pests of export wildflowers and proteas'

25 Two year old C. eatoniae planted in double rows and supported by trellis system. (with insert of detail of trellis system)

Note: Mention of trade names does not imply endorsement of any product by Agriculture Western Australia, and any omission of trade name is unintentional. The support of the wildflower industry and funding from Rural Industries Research and Development Corporation to carry out this work is gratefully acknowledged.

26 Appendix II – Photographs

1. Blue smokebush (C. eatoniae) in a floral arrangement at the FECA ‘Flowerful Australia’ exhibition in Tokyo 1997.

27 2. C. eatoniae in cultivation trials at Medina Research Centre.

3. C. amoenum in cultivation trials at Medina Research Centre.

28 4. In vitro shoot multiplication trials of C. eatoniae.

5. In vitro root growth of C. eatoniae (as viewed from below the plant).