WO2019073443A1

WO2019073443A1 - Composition, system, and method for attracting flying pollinating insects

Info

Publication number: WO2019073443A1
Application number: PCT/IB2018/057906
Authority: WO
Inventors: Ashraf M. El-Sayed; Rikard Unelius
Original assignee: The New Zealand Institute For Plant And Food Research Limited
Priority date: 2017-10-13
Filing date: 2018-10-12
Publication date: 2019-04-18

Abstract

Described are compositions for attracting flying insect pollinators, in particular flying insect pollinators of the orders Lepidoptera, Diptera and Hymenoptera, in particular bumblebees, comprising isophorone and optionally 4-oxoisophorone. Also disclosed are systems and methods for attracting, for attracting and trapping, and for detecting, surveying, monitoring and/or controlling flying pollinating insects.

Description

COMPOSITION, SYSTEM, AND METHOD FOR ATTRACTING FLYING

POLLINATING INSECTS

1. FIELD OF THE INVENTION

The invention provides compositions, systems and methods that can be used to attract flying pollinating insects, in particular flying pollinating insects of the orders Lepidoptera, Diptera and Hymenoptera, in particular bumblebees.

2. BACKGROUND OF INVENTION

Mountain beech forests (Nothofagus spp.) are the largest remaining indigenous forest type in New Zealand, covering a quarter of its land area (ca. 1.2 million ha). These beech forests have become a home to various invasive hymenopteran species including social wasps (two Vespula species) and bumblebees Bombus species, at forest margins. These two invasive insect groups are thriving in association with the beech forest mainly because of the abundant carbohydrate-rich honeydew and the lack of natural enemies.

Bumblebees were brought from England to New Zealand in 1885 by settlers, mainly for red clover pollination. Since their introduction, they have spread all over New Zealand. Four species of bumblebees occur in New Zealand, including the two short-tongued bumblebees Bombus terrestris (Linnaeus) and Bombus subterraneus (Linnaeus), and the two long-tongued bumblebees Bombus ruderatus (Fabricius) and Bombus hortorum (Linnaeus). B. terrestris is the most abundant of the four species.

Bumblebees, especially B. terrestris, are important pollinators of pastoral legumes. They are efficient pollinators of flowers with widely separated styles and stamens, notably kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp) and Brassica (Brassica rapa). The introduction of bumblebees for pollination in New Zealand was the first successful introduction of this genus in the world. This population later acted as a Noah's Ark for the United Kingdom population which had undergone local extinction.

Along with other flying insect pollinators, bumblebees are key components of both natural and agricultural ecosystem. However, disease, pollution and anthropogenic change are threatening many pollinator species, leaving plant-pollinator networks vulnerable. In 1999 the Convention on Biological Diversity issued the Sao Paulo Declaration on Pollinators, in recognition of their crucial role in supporting terrestrial productivity. Consequently, compositions that attract insect pollinators can help monitor plant-pollinator networks, as well as enhancing crop pollination. Improved pollination has a significant impact on the yield and quality of many crops, so pollinator attractants can also help increase crop yields.

It is therefore an object of the invention to provide compositions, systems and methods for attracting flying pollinator insects, in particular, bumblebees, or to at least provide the public with a useful choice.

3. SUMMARY OF THE INVENTION

The inventors have surprisingly discovered that selected compounds present in honey attract bumblebees and other flying insect pollinators. Isophorone (3,5,5-trimethyl-2- cyclohexene-l-one) in combination with 4-oxoisophorone (2,6,6-trimethyl-2-cyclohexene- 1,4-dione) has been found to be particularly efficacious.

Accordingly, in one aspect the invention provides a composition for attracting flying insect pollinators comprising isophorone and optionally 4-oxoisophorone.

In another aspect the invention provides a system for detecting, surveying, monitoring and/or controlling flying insect pollinators comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

In one embodiment, the system comprises a killing agent.

In another aspect the invention provides a system for attracting flying insect pollinators comprising a dispenser which dispenses a vapour blend of a composition comprising isophorone and optionally 4-oxoisophorone.

In another aspect, the invention provides a system for attracting and trapping flying insect pollinators comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device. In another aspect, the invention provides a method for detecting, surveying, monitoring and/or controlling flying pollinating insect populations in an area comprising : placing in the area, a system comprising :

(b) a trapping device.

In another aspect, the invention provides a method of attracting flying pollinating insects to a location, the method comprising placing a composition of the invention at that location.

In various embodiments, the method further comprises trapping one or more of the flying pollinating insects.

In various embodiments, the method comprises placing in the area, a system comprising :

(a) a dispenser which dispenses a vapour blend of a composition of claim 1, and

(b) a trapping device.

In various embodiments, the trapping of the one or more of the flying pollinating insects and/or the trapping device is non-lethal.

In various embodiments, the method further comprises domesticating the one or more trapped flying pollinating insects.

In another aspect, the present invention provides a method for attracting and trapping flying pollinating insects comprising : placing at a location, a system comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

In various embodiments, the trapping device is non-lethal.

In another aspect the invention provides a method for increasing crop yields by attracting flying pollinating insects to the location of the crops, the method comprising placing a composition of the invention at that location. In some embodiments, the crop is selected from the group consisting of fruit and vegetable crops, such as kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp), brassica (Brassica rapa), avocados (Persea americana), apples (Malus domestica), carrots (Daucus carota), onions (Allium cepa), tomatoes (Solanum lycopersicum), strawberry (Fragaria x ananassa), capsicum (Capsicum annum), eggplant (Solanum melongena), legume crops, such as bean

(Phaseolus vulgaris) and pea (Pisum sativum), nut and seed crops, such as almonds (Prunus dulcis), coffee (Coffea spp.), buckwheat (Fagopyrum esculentum), sunflower (Helianthus annuus), macadamia (Macadamia spp. ), pigeonpea (Cajanus cajan), clover (Trifolium repens), annatto (Bixa orellana), fruit crops, such as cherry (Prunus spp.), watermelon (Citrullus lanatus), blueberry (Vaccinnium spp.), mango (Mangifera spp), grapefruit (Citrus x paradisi), longan (Dimocarpus longan), acerola (Malpighia

emarginata), vegetable crops, such as pumpkin (Curcubita pepo), and fibre crops, such as cotton (Gossypium spp) and rapeseed (Brassica napus). In one embodiment the crop is selected from the group comprising kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp) and Brassica (Brassica rapa).

In some embodiments, the crop is grown in an open field. In other embodiments, the crop is grown in a greenhouse.

The following embodiment may relate to any of the above aspects in any combination.

In various embodiments, the composition comprises a synergistic mixture of isophorone and 4-oxoisophorone. In one embodiment, the composition comprises a synergistic vapour mixture of isophorone and 4-oxoisophorone.

In various embodiments, the composition comprises isophorone and 4-oxoisophorone in a wt ratio of about 100: 0 to 10 :90, preferably about 90 : 10 to about 50: 50.

In various embodiments, the wt ratio of isophorone to 4-oxoisophorone may be about 100: 0, 95: 5, 90: 10, 85: 15, 80: 20, 75 : 25, 70 : 30, 65 : 35, 60 :40, 55:45, 50: 50, 45: 55, 40 : 60, 35: 65, 30 : 70, 25: 75, 20 : 80, 15: 85, 10: 90 or 5:95, and useful ranges may be selected between any of these values (for example, about 95: 5 to about 60:40; about 90 : 10 to about 40: 60; about 90 : 10 to about 50: 50).

In various embodiments, the flying pollinating insect is an insect of one of the orders selected from Lepidoptera, Diptera and Hymenoptera. In various embodiments, the flying pollinating insect is of the order Hymenoptera.

Preferably, the flying pollinating insect is a honey bee or bumblebee, more preferably a bumblebee. In certain embodiments, the flying pollinating insect is a bumblebee queen.

In various embodiments, the flying pollinating insect is of the order Diptera. Preferably, the flying pollinating insect is a Tachinidae fly or a Syphid fly.

In various embodiments, the flying pollinating insect is of the order Lepidoptera.

Preferably, the flying pollinating insect is a Magpie moth.

In various embodiments where the flying pollinating insect is of the order Hymenoptera, preferably a honey bee or bumblebee, more preferably a bumblebee, for example a bumblebee queen, the wt ratio of isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to 50:50, 95:0 to 50:50, 90:10 to 50:50, 100:0 to 75:25, 95:5 to 75:25, or 90:10 to 75:25.

In various embodiment where the flying pollinating insect is of the order Diptera, preferably Tachinidae fly or a Syphid fly, the wt ratio of isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to about 25:75, 95:5 to about 25:75, 90:10 to 25:75, 80:20 to 25:75, 75:25 to 25:75, 100:0 to about 40:60, 95:5 to about 40:60, 90:10 to 40:60, 80:20 to 40:60, 75:25 to 40:60, 100:0 to 50:50, 95:0 to 50:50, 90:10 to 50:50, 80:20 to 50:50, or 75:25 to 50:50

In various embodiments where the flying pollinating insect is of the order Lepidoptera, preferably a Magpie moth, the wt ratio of isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to 50:50, 95:0 to 50:50, 90:10 to 50:50, 100:0 to 75:25, 95:5 to 75:25, 90:10 to 75:25.

It is intended that reference to a range of numbers disclosed herein (for example, 1 to 10) also incorporates reference to all rational numbers within that range (for example, 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9 and 10) and also any range of rational numbers within that range (for example, 2 to 8, 1.5 to 5.5 and 3.1 to 4.7) and, therefore, all sub-ranges of all ranges expressly disclosed herein are hereby expressly disclosed. These are only examples of what is specifically intended and all possible combinations of numerical values between the lowest value and the highest value enumerated are to be considered to be expressly stated in this application in a similar manner.

In this specification where reference has been made to patent specifications, other external documents, or other sources of information, this is generally for the purpose of providing a context for discussing the features of the invention. Unless specifically stated otherwise, reference to such external documents is not to be construed as an admission that such documents, or such sources of information, in any jurisdiction, are prior art, or form part of the common general knowledge in the art.

The invention may also be said broadly to consist in the parts, elements and features referred to or indicated in the specification of the application, individually or collectively, in any or all combinations of two or more of said parts, elements or features, and where specific integers are mentioned herein which have known equivalents in the art to which the invention relates, such known equivalents are deemed to be incorporated herein as if individually set forth.

Although the present invention is broadly as defined above, those persons skilled in the art will appreciate that the invention is not limited thereto and that the invention also includes embodiments of which the following description gives examples.

4. BRIEF DESCRIPTION OF THE FIGURES

The invention is now described with reference to the figures in which :

Figure 1 shows the mean ± SE numbers of insects captured in red delta traps baited with six specific honey compounds at the margins of New Zealand Nothofagus beech forests. Figure 1A = bumblebee workers, Figure IB = bumblebee queens and Figure 1C = honeybee workers. The treatments, from left to right, are as follows: (i) benzaldehyde lOOmg; (ii) benzyl alcohol lOOmg; (iii) phenylacetaldehyde lOOmg; (iv) 2-phenylethanol lOOmg; (v) isophorone lOOmg; (vi) o-methoxyacetophenone lOOmg; (vii) negative control. Treatments labelled with the same case letters are not significantly different (P > 0.05). Treatments that caught no bumblebees were not included in the analyses.

Figure 2 shows the mean ± SE numbers of insects captured in red delta traps baited with lures containing different ratios of isophorone and 4-oxoisophorone at the margins of New Zealand Nothofagus beech forests. Figure 2A = bumblebee workers, Figure 2B = bumblebee queens and Figure 2C = honeybee workers. The treatments, from left to right, are as follows: (i) isophorone lOOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) isophorone 50mg and 4-oxoisophorone 50mg; (iv) isophorone lOmg and 4- oxoisophorone 90mg; (v) blank. Treatments labelled with the same case letters are not significantly different (P > 0.05). Treatments that caught no bumblebees were not included in the analyses. Figure 3 shows the mean ± SE numbers of insects captured in red delta traps baited with lures containing two blends of benzaldehyde, benzyl alcohol, phenylacetaldehyde, and 2- phenylethanol and isophorone at the margins of New Zealand Nothofagus beech forests. Figure 3A = bumblebee workers, Figure 3B = bumblebee queens and Figure 3C = honeybee workers. The treatments, from left to right, are as follows: (i) benzaldehyde 25mg, benzyl alcohol 25mg, phenylacetaldehyde 25mg, 2-phenylethanol 25mg, isophorone 90mg, and 4-oxoisophorone lOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) benzaldehyde 25mg, benzyl alcohol 25mg, phenylacetaldehyde 25mg, and 2- phenylethanol 25mg; (iv) honey; (v) blank. Treatments labelled with the same case letters are not significantly different (P > 0.05). Treatments that caught no bumblebees were not included in the analyses.

Figure 4 shows the mean ± SE numbers of insects captured in red delta traps baited with lures containing isophorone and ortho-methoxyacetophenone in a brassica field in 2016. The treatments, from left to right, are as follows: (i) isophorone lOOmg ; (ii) o- methoxyacetophenone lOOmg; (iii) blank. Treatments labelled with the same case letters are not significantly different (P > 0.05). Treatments that caught no bumblebees were not included in the analyses. Black bars = Lepidoptera; light grey bars = Syrphidae; dark grey bars = other Diptera.

Figure 5 shows the mean ± SE numbers of insects captured in red delta traps baited with lures containing different ratios of isophorone and 4-oxoisophorone in brassica field in

2016. The treatments, from left to right, are as follows: (i) isophorone lOOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) isophorone 50mg and 4-oxoisophorone 50mg; (iv) isophorone lOmg and 4-oxoisophorone 90mg; (v) blank. Treatments labelled with the same case letters or characters are not significantly different (P > 0.05).

Treatments that caught no insects were not included in the analyses. Black bars =

Lepidoptera; light grey bars = Syrphidae; dark grey bars = other Diptera.

Figure 6 shows the mean ± SE numbers of insects captured in red delta traps baited with lures containing different ratios of isophorone and 4-oxoisophorone in a brassica field in

2017. The treatments, from left to right, are as follows: (i) isophorone lOOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) isophorone 50mg and 4-oxoisophorone 50mg; (iv) isophorone lOmg and 4-oxoisophorone 90mg; (v) blank. Treatments labelled with the same case letters or characters are not significantly different (P > 0.05).

Lepidoptera; light grey bars = Syrphidae; dark grey bars = other Diptera. Figure 7 shows the mean ± SE numbers of Tachinidae insects captured in red delta traps baited with lures containing different ratios of isophorone and 4-oxoisophorone in a brassica field. The treatments, from left to right, are as follows: (i) isophorone lOOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) isophorone 50mg and 4-oxoisophorone 50mg; (iv) isophorone lOmg and 4-oxoisophorone 90mg; (v) blank. No significant difference was observed between treatments (P > 0.05).

Figure 8 shows the mean ± SE numbers of Magpie moth insects captured in red delta traps baited with lures containing different ratios of isophorone and 4-oxoisophorone in a pine forest. The treatments, from left to right, are as follows: (i) isophorone lOOmg; (ii) isophorone 90mg and 4-oxoisophorone lOmg; (iii) isophorone 50mg and 4-oxoisophorone 50mg; (iv) isophorone lOmg and 4-oxoisophorone 90mg; (v) blank. Treatments labelled with the same case letters or characters are not significantly different (P > 0.05).

Treatments that caught no insects were not included in the analyses.

5. DETAILED DESCRIPTION OF THE INVENTION

5.1 Definitions

The term 'comprising' as used in this specification and claims means 'consisting at least in part of . When interpreting statements in this specification and claims which include the term 'comprising', other features besides the features prefaced by this term in each statement can also be present. Related terms such as 'comprise' and 'comprised' are to be interpreted in similar manner.

The term "crop" refers to plants which are grown by humans for various purposes such as, but not limited to, obtaining food or any other material from the plant or plant parts including products such as oils, carbohydrates, medicines; ornamental purposes;

landscaping purposes. Crops may be grown in a field, gardens, greenhouses or any other place, on a large or small scale.

The term "pollinator" means an animal that moves pollen from the male anther of a flower to the female stigma of a flower, which helps bring about fertilisation of the ovules of the flower by the male gametes in the pollen grains. "Flying insect pollinators" and "flying pollinating insects" are flying insects whose behaviour results in pollination of one of more species of plant. The terms "flying pollinating insect" and "flying insect pollinator" are used interchangeably herein.

As used herein "(s)" following a noun means the plural and/or singular forms of the noun. As used herein the term "and/or" means "and" or "or" or both.

5.2 The compositions of the invention and their uses

Isophorone (CAS registry 78-59-1) and 4-oxoisophorone (CAS registry 1125-21-9) are norisoprenoids present in honey. The inventors have surprisingly discovered that isophorone attracts flying pollinator insects and acts synergistically when combined with 4- oxoisophorone.

Accordingly, in one aspect the invention provides a composition for attracting flying insect pollinators comprising isophorone and optionally 4-oxoisophorone. In one embodiment, the composition comprises a synergistic mixture of isophorone and 4-oxoisophorone. In one embodiment, the composition comprises a vapour mixture of isophorone and 4- oxoisophorone.

The ratio of isophorone to 4-oxoisophorone may be about 100: 0, 95: 5, 90: 10, 85 : 15, 80 : 20, 75: 25, 70: 30, 65: 35, 60 :40, 55 :45, 50: 50, 45 : 55, 40: 60, 35: 65, 30: 70, 25 : 75, 20 : 80, 15: 85, 10:90 or 5:95, and useful ranges may be selected between any of these values (for example, about 95 : 5 to about 60:40; about 90: 10 to about 40: 60; about 90 : 10 to about 50: 50). In one embodiment, the composition comprises isophorone and 4-oxoisophorone in a wt ratio of about 100 : 0 to 10:90, preferably about 90 : 10 to about 50 : 50.

The most effective ratio of isopharone to 4-oxoisophorone may vary depending on the target pollinator and other conditions. A person skilled in the art could readily determine the optimum composition for the particular purpose.

The compositions of the invention may also comprise carriers, such as solvents (eg, ether, alcohol, acetone, hexane and the like) or oils. Water is generally not a very suitable carrier, as the active agents are not very water soluble. The composition may also comprise other ingredients such as nutrients, diluents, emulsifiers, wetting agents, surfactants, dispersants, stabilisers and the like. The other components should have no negative biological effects on plants.

The percentage of active agents isophorone and 4-oxoisophorone in the composition of the invention may vary depending on the activity of the composition towards the target insect species, the type of formulation, the site and mode of application. The percentage of active agents in the composition of the invention may at about 1%, 2%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 99% or even 100% wt/wt or wt/vol of the composition.

(b) a trapping device.

In one embodiment, the system comprises a killing agent.

In another aspect, the invention provides a system for attracting and trapping flying insect pollinators insects comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

The systems of the invention dispense the composition of the invention into the

surrounding air where it is detected by the insect. In one embodiment, the dispenser dispenses a vapour blend of the composition of the invention. In some embodiments, the dispenser dispenses an aerosol of the composition of the invention.

A variety of devices have been described that can evaporate or dispense volatile compounds and compositions into the atmosphere; see for example, US 6,582,714 and US 7,481,380. The dispenser may be active or passive. Active dispensers include aerosol sprayers that propel minute droplets of composition into the air. These may be operated by pressure, air displacement or pump action. Dispensers may also utilise heating or burning to evaporate the attractant composition. Other dispensing methods include substrates such as paper or fabrics impregnated with volatile active ingredients. Evaporative surface dispensers may use a wick or porous surface that provides a large surface area from which the volatile liquid can more quickly evaporate passively in the air. Evaporative surfaces may be provided by materials such as cotton, paper, textiles, and polymeric matrices in the form of plugs or pellets.

In other embodiments, the dispenser may constitute an impermeable membrane or wall that includes a small portion of permeable membrane or one or more small openings. The composition of the invention or its components are contained within the impermeable membrane or wall, and enter the atmosphere via the permeable membrane or small openings. Examples of suitable dispensers are described in Leonhardt et al. (Insect Pheromone Technology: Chemistry and Applications, ACS Symposium Series 90, 1982, incorporated by reference).

The dispenser for use in the systems of the invention may contain or hold the liquid compounds which together comprise the composition of the invention, and also dispense these as a vapour blend. Alternatively, the dispenser may hold and release the liquid compounds separately which then form a vapour blend when they meet on release.

In one embodiment, the systems of the invention also includes a sex pheromone, or kairomone. The sex pheromone or kairomone may be present in the dispenser, or could be released from another part of the system.

As well as the rate at which the composition of the invention is released into the atmosphere, factors such as the insect population density, age distribution of the population, temperature and wind velocity may all influence the numbers of insects attracted and/or trapped in the methods of the invention. Therefore, the rate of release can be adjusted to account for these external factors.

The rate of release of the composition of the invention can be adjusted by adjusting the dispenser for example, by allowing a bigger surface area to be exposed to the

atmosphere. Factors such as temperature, and wind velocity will influence the release rate. The configuration of the system range can be readily determined by a dose response field test. Where the system of the invention is used for detecting, surveying, monitoring and/or controlling flying pollinating insects, it includes a trapping device. The trapping device can be any suitable device including but not limited to a sticky trap, unitrap, bucket trap and the like. In one embodiment, the trapping device is a sticky trap, such as a Delta trap.

In use, the insects are attracted to the composition dispensed from the system and are then trapped in the trapping device, allowing them to be counted. Trapped insects can either be disposed of, if still alive when the system is checked, or the system can include a killing agent. In some cases the trapping device will constitute a killing agent.

Killing agents include, but are not limited to, insecticides, soapy water and the like.

In one embodiment, the killing agent is a carbamate, organophosphorous compound, nitrophenol, nitromethylene, phenylbenzoylurea, pyrethroid, chlorinated hydrocarbon or microbial insecticide. Preferably, the killing agent is a pyrethroid insecticide.

In the above methods, systems of the invention can be widely distributed to detect the presence of the insects in a particular area. Insect populations can be surveyed by applying mathematical analyses to the results. Changes in populations over time can also be monitored by comparing results with earlier data. If initial results warrant it, mass trapping programmes can be initiated, to control population numbers.

Systems of the invention to be used for attracting insects to crops for pollination do not include trapping devices and merely dispense the composition of the invention.

Systems of the invention to be used for attracting and trapping flying insect pollinators comprise a trapping device. As described herein with respect to the system for detecting, surveying, monitoring and/or controlling flying pollinating insects, the trapping device may be any suitable device. The trapping device may be lethal or non-lethal, depending on the purpose for trapping the insects. In some embodiments, a lethal trapping device may be used, for example, to control of an insect population in an area. In some embodiments, a non-lethal trapping device may be used for trapping one or more insects to be

domesticated, for example bumblebees, such as bumblebee queens, or other domesticable flying pollinating insects. The non-lethal trapping device traps the one or more insects without killing them, and preferably without harming them. Accordingly, in some embodiments, the methods of the invention comprise trapping one or more flying pollinating insects and domesticating the one or more trapped insects.

Other compounds and materials may be added to the composition or system of the invention provided they do not substantially interfere with the release of the active agents, or their attractant properties. Whether or not this occurs can be determined by standard test formats comparing the efficacy of the composition or system and without the added compound or material.

In the trials described in the Examples, the compositions of the invention were placed inside a permeable polyethylene bag (100 μηη wall thickness, 20 mm x 20 mm, Masterton, New Zealand) with a piece of felt (15 mm x 45 mm) inserted as a carrier material. The polyethylene sachets were placed in the centre of the sticky base.

Pollinating insects include but are not limited to bees, flies, moths, butterflies, wasps and ants. The compositions and systems of the invention are effective attractants for flying pollinating insects.

In one embodiment, the flying pollinating insect is an insect of one of the orders selected from Lepidoptera, Diptera and Hymenoptera.

In one embodiment the flying pollinating insect is of the order Hymenoptera, more preferably a bumblebee (Bumbus terrestris), for example a bumblebee queen .

In one embodiment the flying pollinating insect is of the order Lepidoptera, preferably a Magpie moth {Nyctemera annulata) or Red Admiral butterfly {Vanessa gonerilla).

In one embodiment the flying pollinating insect is of the order Diptera, preferably a Tachinidae fly or a Syphid fly.

In another aspect the invention provides a method for increasing crop yields by attracting flying pollinating insects to the location of the crops, the method comprising placing a composition of the invention at that location. The composition may be placed within the area in which the crops are planted, or near to the area.

The compositions of the invention can be used to increase the yield of any crop that is affected by pollination rates.

Examples of suitable crops include but are not limited to fruit and vegetable crops, such as kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp), brassica (Brassica rapa), avocados (Persea americana), apples (Malus domestica), carrots (Daucus carota), onions (Allium cepa), tomatoes (Solanum lycopersicum), strawberry (Fragaria x ananassa), capsicum (Capsicum annum), eggplant (Solanum melongena), legume crops, such as bean (Phaseolus vulgaris) and pea (Pisum sativum), nut and seed crops, such as almonds (Prunus dulcis), coffee {Coffea spp.), buckwheat (Fagopyrum esculentum), sunflower (Helianthus annuus), macadamia

(Macadamia spp.), pigeonpea (Cajanus cajan), clover (Trifolium repens), annatto (Bixa orellana), fruit crops, such as cherry (Prunus spp.), watermelon (Citrullus lanatus), blueberry (Vaccinnium spp. ), mango (Mangifera spp), grapefruit (Citrus x paradisi), longan (Dimocarpus longan), acerola (Malpighia emarginata), vegetable crops, such as pumpkin (Curcubita pepo), fibre crops, such as cotton (Gossypium spp) and rapeseed (Brassica napus), and the like. In one embodiment the crop is selected from the group comprising kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp) and Brassica (Brassica rapa).

The following non-limiting examples are provided to illustrate the present invention and in no way limit the scope thereof.

6. EXAMPLES

Chemicals

The compounds used in field trials: benzaldehyde (98%), benzyl alcohol (99%), phenylacetaldehyde (90%), 2-phenylethanol ( 5^99%), isophorone = 3,5,5- trimethylcyclohex-2-enone (97%), 4-oxoisophorone = 2,6,6-trimethylcyclohex-2-ene-l,4- dione (98%), and ortho-methoxyacetophenone (99%) were obtained from Sigma-Aldrich Chemical Company (St Louis, Missouri).

Field Experimental Protocol for Examples 1 - 3

Field experiments were conducted on a matagouri-covered riverbed adjacent to a mountain beech forest in Hawdon Valley near Arthur Pass in Canterbury, the South Island, New Zealand (42°59.54'S, 171°47.60 E). Red delta traps made of plastic corflute with an adhesive-coated base (Clare et al. 2000) were used. Traps baited with different compounds in five replicates were assigned in five rows, each containing treatments tested in a randomized block design. Traps were positioned 1.7 m above the ground in the matagouri vegetation, and were spaced 20 m apart in each row. The attractant blends were formulated in a permeable polyethylene bag (100 μηη wall thickness, 20 mm x 20 mm, Masterton, New Zealand) of 100 μηη wall thickness (45 mm x 50 mm), with a piece of felt (15 mm x 45 mm) inserted as a carrier substrate. The polyethylene sachets were placed in the center of the sticky base. Sticky bases were checked and insects were identified and counted during the experimental period. Although various other insect species were caught in the trapping trials, only the catch data of bumblebee workers, queen and honeybee workers is presented . In all trials, traps with a dispenser without chemicals was used as the negative control .

Data Analysis

The va riance of mean captures obtained with each compound or each blend of compounds was stabilized using the V (x + 1) transformation of counts and the significance of treatment effects tested using ANOVA. Significantly different treatment means were identified using Fisher's protected least significant difference test (SAS Institute Inc, 1998) .

Example 1: Testing individual compounds as attractants for bumblebees

This field trial, conducted from 25 to 27 February 2012, investigated the relative attractiveness of individual compounds found in honey. Lures of benzaldehyde, benzyl alcohol, phenylacetaldehyde, 2-phenyethanol, isophorone and ortho- methoxyacetophenone were made by dispensing 100 mg of each compound on a piece of felt inside a permeable polyethylene bag that was then heat-sealed . As shown in Figure 1, significantly more bumblebee workers were attracted to isophorone and ortho- methoxyacetophenone, than to the other test compounds or the negative control

(Treatment, F3, 16 = 0.16, P = 0.92) . Isophorone significantly attracted more bumblebee queens and honeybee workers than other tested lures and negative control (bumblebee queen : Treatment, F3, 16 = 0.16, P = 0.92, honeybee worker: Treatment, Fl,8 = 5.6, P = 0.046) .

Example 2: Optimizing the ratio of isophorone (Ip) and 4-oxoisophorone (OIp) as bumblebee attractants

In the second trapping trial ( 1-3 March 2012), the relative attractiveness of various binary blends containing various ratios (100 : 0, 10 : 90, 50 : 50, 10 : 90, and 0 : 100 mg) of isophorone and 4-oxoisophorone was investigated .

Changing the ratio between isophorone and 4-oxoisophorone significantly affected the number of bumblebee workers captured (Fig ure 2) . Bumblebee workers : Treatment, F3,ie = 0.16, P = 0.92, honeybee queens : Treatment, F.,s = 5.6, P = 0.046. The largest number of bumblebee workers caught in traps baited with the bina ry blend at ratio of 90 : 10 (Fig 2) . The catch of bumblebee workers in traps baited with isophorone alone, and binary blends of isophorone and 4-oxoisophorone at ratios of 50 : 50, and 10 : 90 was not significantly different (Fig . 2) . The largest number of bumblebee queens caught in traps baited with the binary blend at ratio of 90 : 10 (Fig 2) . There was no significance difference in honeybee workers caught between treatments (Fig 2) . Example 3: Investigating synergistic effects of benzene derivatives

In the final trapping trial (8-10 March 2012), the relative attractiveness of three blends was investigated. Blend 1 contained benzaldehyde, benzyl alcohol, phenylacetaldehyde, 2- phenylethanol, isophorone, and 4-oxoisophorone at ratio 25: 25 : 25: 25: 90: 10 mg, respectively. Blend 2 contained isophorone, and 4-oxoisophorone at ratio 90 : 10 mg) respectively. Blend 3 contained benzaldehyde, benzyl alcohol, phenylacetaldehyde, 2- phenylethanol at ratio (25 : 25: 25: 25 mg) respectively. Traps baited with 1 gram of clover honeydew honey were used as positive controls.

The addition of benzaldehyde, benzyl alcohol, phenylacetaldehyde, and 2-phenylethanol to a binary blend of isophorone and 4-oxoisophorone did not result in a significant increase in the catch of bumblebee or honeybee workers (Figure 3). In contrast, the addition of these compounds to the binary blend resulted in a significance decrease in the number bumblebee queens caught (Figure 3). The catch of bumblebee or honeybee workers and bumblebee queens in a binary blends containing isophorone and 4-oxoisophorone was similar to the catch in traps baited with honey (Figure 3).

Example 4: Testing individual compounds as attractants for Lepidoptera and Diptera

This trial was conducted in a brassica field from 23 March to 12 April 2016 to investigate the relative attractiveness of the individual compounds. Lures of isophorone, ortho- methoxyacetophenone were made by dispensing 100 mg of each compound on a piece of felt inside a permeable polyethylene bag that was then heat-sealed. Otherwise, the protocol was the same as for Examples 1-3.

Significantly more syrphid flies were caught in traps baited with isophorone than in traps baited with o-methoxyacetophenone (Figure 4).

Example 5: Optimizing the ratio of isophorone and 4-oxoisophorone for syrphid flies

In a further trial (1-3 March 2012), in beech forest margin the relative attractiveness of various binary blends containing various ratios (100: 0, 10:90, 50: 50 and 10:90 mg) of isophorone and 4-oxoisophorone was investigated.

In addition, this trial was repeated in 2016 (12 Mar 15 to 12 April) 2017 (1 Apr to 15 Apr) in brassica field (43°39'49.9 S; 171°55'55.2 E) . In trials conducted in brassica field, a large number of syrphid flies were caught in traps baited with either single component or a binary blend of the two components. In the trial conducted in 2016, significantly large number of syrphid flies were caught in a blend containing isophorone alone or isophorone and 4-oxoisophorone at a ratio of (10:90) (Figure 5). Significantly more Lepidoptera and other Diptera were caught in traps baited with all synthetic blend than in blank control (Figure 5). In the second trial conducted in 2017, significantly more syrphid flies and other Diptera were caught in traps baited with a blend of isophorone and 4-oxoisophorone at a ratio of 50: 50 and 10 :90 (Figure 6).

Example 6: Isophorone and 4-oxoisophorone as attractant for Tachinidae flies and magpie moth

In a trapping trial (10-21 Feb 2015), the relative attractiveness of various binary blends containing various ratios (100 : 0, 90 : 10, 50 : 50 and 10 :90 mg) of isophorone and 4- oxoisophorone was investigated in a brassica field. In this trial, changing the ratio between isophorone and 4-oxoisophorone affected the number of Tachinidae flies captured (Figure 7). No flies were captured in the blank trap.

In an earlier trapping trial (10-27 Feb 2013), the relative attractiveness of various binary blends containing various ratios (10: 0, 10:90, 50: 50 and 10 :90 mg) of isophorone and 4- oxoisophorone was investigated in pine forest in Canterbury, South Island, New Zealand. Changing the ratio between isophorone and 4-oxoisophorone significantly affected the number of magpie moth, Nyctemera annulata (Boisduval) captured (Figure 8). The largest number of magpie moth were caught in traps baited with isophorone alone and with a binary blend at a ratio of 90: 10 (Figure 8).

7. INDUSTRIAL APPLICABILITY

The compositions, systems, and methods are useful for various purposes as would be appreciated by those skilled in the art. As described herein, the compositions, systems, and methods are useful for attracting flying insect pollinators, for example to trap or capture such insects or to enhance the pollination of crops by such insects.

In certain exemplary embodiments, the compositions, systems, and methods are useful for attracting bumblebees. The ecological impact of the large earth bumble bee populations adjacent to the New Zealand mountain beech forests may have been overlooked in the shadow of the vast ecological catastrophe imposed by the predatory Vespula wasps. Potentially, New Zealand native insect species that specifically feed on the same food resource as the bumble bees may locally be in competition. The identification of a potent bumblebees attractants will enable their population to be monitored in the mountain beech forests across New Zealand .

In addition, these attractants could also be used to manage bumble bees in recreation areas where large number of bumble bees has led to an increasing number of humans getting stung accidentally. These attractants could also be used to enhance crop pollination by attracting bumblebees and other pollinators to the right place.

8. REFERENCE

Clare G, Suckling DM, Bradley SJ, Walker JTS, Shaw PW, Daly JM, McLaren GF, Wearing CH. (2000) Pheromone trap colour determines catch of non-target insects. New Zealand Plant Protection 53: 216-220.

Any section headings used herein are for organizational purposes only and are not to be construed as limiting the subject matter described.

Although the invention has been described by way of example and with reference to particular embodiments, it is to be understood that modifications and/or improvements may be made without departing from the scope of the invention.

Claims

1. A composition for attracting flying insect pollinators comprising isophorone and

optionally 4-oxoisophorone.

2. A system for detecting, surveying, monitoring and/or controlling flying insect

pollinators comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

3. The system of claim 2, wherein the system comprises a killing agent.

4. A system for attracting flying insect pollinators comprising a dispenser which dispenses a vapour blend of a composition comprising isophorone and optionally 4- oxoisophorone.

5. A system for attracting and trapping flying insect pollinators comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

6. A method for detecting, surveying, monitoring and/or controlling flying pollinating insect populations in an area comprising : placing in the area, a system comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

7. A method of attracting flying pollinating insects to a location, the method comprising placing a composition of claim 1 at that location.

8. The method of claim 7, wherein the method further comprises trapping one or more of the flying pollinating insects.

9. A method for attracting and trapping flying pollinating insects comprising : placing at a location, a system comprising :

(a) a dispenser which dispenses a vapour blend of a composition comprising

isophorone and optionally 4-oxoisophorone, and

(b) a trapping device.

10. The method of claim 9, wherein the trapping device is non-lethal.

11. The method of any one of claims 8-10, wherein the method further comprises domesticating the one or more trapped flying pollinating insects.

12. A method for increasing crop yields by attracting flying pollinating insects to the

location of the crops, the method comprising placing a composition of claim 1 at that location.

13. The method of claim 12, wherein the crop is selected from the group the crop is

selected from the group consisting of fruit and vegetable crops, such as kiwifruit (Actinidia chinesis), passion fruit (Passiflora edulis), feijoa (Acca sellowiana), cucurbits (Cucurbita spp), brassica (Brassica rapa), avocados (Persea americana), apples (Malus domestica), carrots (Daucus carota), onions (Allium cepa), tomatoes (Solanum lycopersicum), strawberry (Fragaria x ananassa), capsicum (Capsicum annum), eggplant (Solanum melongena), legume crops, such as bean (Phaseolus vulgaris) and pea (Pisum sativum), nut and seed crops, such as almonds (Prunus dulcis), coffee (Coffea spp.), buckwheat (Fagopyrum esculentum), sunflower (Helianthus annuus), macadamia (Macadamia spp.), pigeonpea (Cajanus cajan), clover (Trifolium repens), annatto (Bixa orellana), fruit crops, such as cherry (Prunus spp.), watermelon

(Citrullus lanatus), blueberry (Vaccinnium spp.), mango (Mangifera spp), grapefruit (Citrus x paradisi), longan (Dimocarpus longan), acerola (Malpighia emarginata), vegetable crops, such as pumpkin (Curcubita pepo), and fibre crops, such as cotton (Gossypium spp) and rapeseed (Brassica napus).

14. The composition, system, or method of any one of the preceding claims, wherein the composition comprises a synergistic mixture of isophorone and 4-oxoisophorone.

15. The composition, system, or method of any one of the preceding claims, wherein the composition comprises a synergistic vapour mixture of isophorone and 4- oxoisophorone.

16. The composition, system, or method of any one of the preceding claims, wherein the composition comprises isophorone and 4-oxoisophorone in a wt ratio of about 100 : 0 to 10 :90.

17. The composition, system, or method of claim 16, wherein the wt ratio of isophorone to 4-oxoisophorone is from about 90: 10 to about 50: 50.

18. The composition, system, or method of any one of the preceding claims, wherein the wt ratio of isophorone to 4-oxoisophorone is about 100 : 0, 95 : 5, 90: 10, 85: 15, 80 : 20, 75 : 25, 70: 30, 65: 35, 60:40, 55:45, 50 : 50, 45: 55, 40 : 60, 35: 65, 30: 70, 25: 75, 20 : 80, 15: 85, 10:90 or 5:95.

19. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is an insect of one of the orders selected from Lepidoptera, Diptera and Hymenoptera.

20. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is of the order Hymenoptera.

21. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is a honey bee or bumblebee.

22. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is a bumblebee.

23. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is a bumblebee queen.

24. The composition, system, or method of any one of claims 21-23, wherein the wt ratio of isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to 50:50, 95:0 to 50:50, 90:10 to 50:50, 100:0 to 75:25, 95:5 to 75:25, or 90:10 to 75:25.

25. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is of the order Diptera.

26. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is a Tachinidae fly or a Syphid fly.

27. The composition, system, or method of claim 25 or 26, wherein the wt ratio of

isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to about 25:75, 95:5 to about 25:75, 90:10 to 25:75, 80:20 to 25:75, 75:25 to 25:75, 100:0 to about 40:60, 95:5 to about 40:60, 90:10 to 40:60, 80:20 to 40:60, 75:25 to 40:60, 100:0 to 50:50, 95:0 to 50:50, 90:10 to 50:50, 80:20 to 50:50, or 75:25 to 50:50.

28. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is of the order Lepidoptera.

29. The composition, system, or method of any one of the preceding claims, wherein the flying pollinating insect is a Magpie moth.

30. The composition, system, or method of claim 28 or 29, wherein the wt ratio of

isophorone to 4-oxoisophorone is from about 100:0 to about 5:95, 95:5 to 5:95, 100:0 to 10:90, 95:5 to 10:90, 90:10 to 10:90, 100:0 to 50:50, 95:0 to 50:50, 90:10o 50:50, 100:0 to 75:25, 95:5 to 75:25, 90:10 to 75:25.