BOLLGARD 3 REFUGE PLANNING GUIDE GET THE MOST OUT OF YOUR REFUGE

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1 BOLLGARD 3 REFUGE PLANNING GUIDE GET THE MOST OUT OF YOUR REFUGE Refuge crops are part of the Australian cotton industry s resistance management plan to proactively preserve leading insect resistance technology. It is critical that refuges are well planned and managed to ensure technology preservation for the future and it s important that growers are aware of the importance of their obligations. Australian cotton growers are the first Bollgard 3 cotton growers in world. Bollgard 3 provides a more flexible resistance management plan, with reduced refuge area, allowing for greater areas of productive cotton to be produced. Growers have a critical role to play to implement the Bollgard 3 Resistance Management Plan (RMP) on their farm, including planting effective refuge crops. Effective refuges are critical to the longevity of the leading insect technology in Australia. Dr Kristen Knight, Monsanto Entomologist

2 What are some of the refuge challenges faced by growers? Poor pigeon pea stand due to cool (and often wet) conditions during establishment and poor seedling vigor. Late replanting of pigeon pea crops resulting in a lack of flowering synchronization with Bollgard 3 cotton, thus reducing Helicoverpa attractiveness. Control of Ratoon and volunteer cotton within refuge crops - while this has improved this is an area where continued vigilance is required. Weed control in Pigeon Pea refuges. Moisture stress in refuge crops relative to the Bollgard 3 cotton. Avoiding spraying of refuges with Bt insectides. Even when accidental, this can render the refuge ineffective. The importance of a wellplanned refuge in maximising effectiveness Refuge Rules of Thumb Refuges must be: healthy and either Pigeon Pea or Unsprayed conventional cotton. planted and managed so that they are attractive to Helicoverpa spp. during the entire growing period of the Bollgard 3 cotton. planted within specific windows relative to Bollgard 3 cotton for dryland and irrigated crops (refer to page 9 for details). non-moisture stressed for the entirety of the cotton field s growing season relative to the health of Bollgard 3 cotton. planted within the farm unit growing the Bollgard II and Bollgard 3 cotton. If cultivation is required in the refuge, the Bollgard 3 cotton should also be cultivated at the same time. Once flowering commences in the Bollgard 3 cotton the refuge must not be cultivated. All reasonable effort should be taken to plant the refuge adjacent to Bollgard 3 cotton fields. All Bollgard 3 fields must be within 2 km from the nearest pigeon pea refuge crop. If fields are Bollgard 3/Roundup Ready Flex or Roundup Ready Flex, up to 200 metres of buffer between the crops can be left to prevent herbicide drift. The minimum refuge size is 0.5 Hectares and 2 Hectares if there is any sprayed conventional cotton on the farm unit. Refuge Planting How close does my refuge sowing date need to be to my Bollgard 3 cotton? If refuges are planted outside of the recommended RMP windows it is unlikely that they will be attractive at the same time as your Bollgard 3 cotton crop. (Refer to Tables 1, 2, 4 & 5 for refuge requirements and planting windows). The aim of the refuge is to produce large numbers of susceptible moths during the period when moths could emerge from Bollgard 3 cotton. This cannot happen if your refuge is not attractive/flowering at the same time as the Bollgard II crop. Irrigated: It is preferable that all refuge is planted within the 2-week period prior to planting Bollgard 3. If this is not possible, refuge planting must be completed within 3 weeks of the first day of sowing of Bollgard 3. Dryland: A dryland refuge must be planted within the 2-week period prior to the first day of planting Bollgard 3 cotton. Dryland pigeon pea refuges must be approved by your local Monsanto RBM prior to planting. For further information on general conditions for refuges refer to the Bollgard 3 Resistance Management Plan: wp-content/uploads/2015/10/bollgard-3- Resistance-Management-Plan.pdf

3 Refuge Distances How close to Bollgard 3 cotton does a refuge have to be? Why 2km? The distance that a refuge is planted from Bollgard 3 cotton is critical for resistance management based on assumptions behind the theory of gene flow during mating. Resistance genes are considered rare and refuge placement is critical in ensuring susceptible moths dominate populations. This is achieved by ensuring that the large majority of resistant insects (RR) that survive in the Bollgard 3 environment mate with insects reared on plants that do not contain Bt, thus are likely to be homozygous susceptible (SS). All growers are required to grow a refuge crop that can produce large numbers of Helicoverpa spp. moths which have not been exposed to Bollgard 3 proteins. These will dominate matings with any survivors from Bollgard 3 crops and maintain resistance at low levels. Gary Fitt (CSIRO), Peter Gregg (UNE and Cotton CRC) and colleagues have undertaken dispersal studies of H. armigera. Findings have shown that there is an exponential decrease in recaptured marked moths as distance increases from the source of the marked moths i.e. the area they were released from. This is represented in Figure 1. BOLLGARD 3 25 % BOLLGARD % BOLLGARD % moth movement from centre of one field through to the other 1 km moth movement from centre of one field through to the other 2 km REFUGE REFUGE REFUGE moth movement from centre of one field through to the other Figure 1. Helicoverpa spp moth movement from refuge to Bollgard 3 cotton field relative to distance. Why is refuge width important? The aim of a refuge is to produce large numbers of susceptible insects. Therefore, the resistance management plans for Bollgard 3 cotton require for refuges to be a minimum of 24m wide and 0.5 Hectare in size. Refuge dimensions are put in place to ensure that a refuge will not be affected by potential insecticide drift. It is also to encourage canopy density, making it attractive for moths to lay their eggs within the refuge. Studies have found that egg densities are significantly higher in wider refuges (Caprio et al, 2004).

4 Unsprayed Pigeon Pea Refuges for Bollgard 3 Cotton Key advantages of a pigeon pea refuge Unsprayed pigeon pea has the ability to generate twice the number of susceptible moths as the same area of unsprayed cotton when used as a refuge 50% of the refuge area required compared to the same area of unsprayed cotton refuges (refer to Tables 1 & 2 for Bollgard 3 areas) Independent research continues to demonstrate the effectiveness of Pigeon Pea as a refuge Pigeon Peas are a legume and have nitrogen fixing capacity in soils where low residual nitrogen exists. Pigeon Pea Bollgard 3 Cotton Resistance Management Plan Requirements Pigeon Pea refuge area requirements Crop Condition % of Bollgard 3 Central Queensland If planted between August 1 October 31 If planted between November 1 December 31 Irrigated & Dryland* Irrigated & Dryland* Pigeon Pea Unsprayed pigeon peas 2.5% 5% *Subject to approved management plan by Monsanto Table 1. Central Queensland Bollgard 3 RMP pigeon pea requirements

5 Crop Condition % of Bollgard 3 Southern Queensland, NSW and Victoria All cotton planted between August 1 December 31 Irrigated & Dryland* Pigeon Pea Unsprayed pigeon peas 2.5% *Subject to approved management plan by Monsanto Table 2. Southern Queensland, NSW and Victoria Bollgard 3 RMP pigeon pea requirements. Control of Volunteer and Ratoon cotton Volunteer and ratoon cotton, within fields planted back to back may impose additional selection pressure on Helicoverpa spp. to develop resistance to the Bollgard 3 proteins. Therefore, it is critical that all fields planned for pigeon pea sowing are clean of Bollgard 3 volunteer or ratoon cotton prior to planting. The easiest way to avoid this is to ensure that refuges are not sown into last years Bollgard 3 stubble. Pigeon pea refuges must be kept clean of volunteer and ratoon cotton for the entire growing period. If ratoon Bollgard 3 cotton is left to persist Helicoverpa spp. larvae could develop beyond the third instar on the refuge and move onto the volunteer Bollgard 3 cotton (Figure 1). This poses a high risk for resistance development. Pupae Destruction in Pigeon Pea Refuges To ensure maximum emergence of late pupae from the pigeon pea refuge, soil disturbance in the refuge should not occur until after the pupae busting in the Bollgard 3 cotton is complete. Ideally, the pigeon pea refuge should be left uncultivated until the following spring if possible. Pigeon pea refuges do not have to be cultivated. Remember the more susceptible moths that emerge from the refuge the greater the genetic dilution of any resistant survivors. Direct drilling winter crops into pigeon pea refuge is permitted. Pigeon Pea harvest or crop destruction Harvest or crop destruction of aerial parts of the pigeon pea refuge can only occur after the Bollgard 3 cotton lint has been removed.

6 Pigeon Pea Agronomic Considerations Simple Steps to produce an effective Pigeon Pea Refuge Field Selection: Avoid planting pigeon pea into last year s Bollgard 3 cotton field. Where possible plant pigeon pea into well drained soils; pigeon peas are less tolerant to waterlogging than cotton. Planting Seed Quality: Always perform a germination test prior to sowing and or purchase QA seed. Adjust the sowing rate to ensure a plant stand equivalent to the Bollgard 3 cotton is achieved. Inoculate pigeon pea seed to maximise nitrogen fixation opportunities within the rotation. Certified seed is always preferred to ensure variety purity, which will improve synchrony with the Bollgard 3 cotton flowering period. If saving seed from the previous pigeon pea crop, do not use glyphosate as a desiccant, if seed maturity is not complete, then seed viability is likely to be affected. Weed management: Apply a residual herbicide for long term weed control. Prometryn, Trifluralin and Sencor are all registered. Remember don t let this year s refuge generate next year s weeds.

7 Active Ingredient Mode of Action Concntration and formulation Application rate of product Comment Prometryn* Trifluralin C D 500 g/l CS Apply up to 4.5 L/ha Apply up to the maximum rate pre planting and incorporate, or as a post-emergent directed spray towards the base of establishment 900 g/kg WDG Apply up to 2.5 L/ha plantss (Per13758) 480 g/l EC* Apply up to 2.3 L/ha Apply up to the maximum rate pre planting and incorporate 530 g/l EC Apply up to 1.5 L/ha NSW and ACT only 600 g/l EC Apply up to 1.4 L/ha NSW and ACT only Butroxydim* A 250 g/l WDG Apply 180 g/ha 212 g/l EC Apply 1 L/ha Fluazifop-p* A 128 g/l EC Apply 1.6 L/ha Haloxyfop* A 130 g/l Apply 0.6 L/ha Apply the specified rate as a post-emergence spray over the top of the pigeon pea crops. Refer to label as rates are different depending on weed being controlled. (Per13758) Haloxyfop* A 520 g/l EC Apply L/ha Sethoxydim* A 186 g/l EC Apply 1 L/ha Clethodim* A 240 g/l EC Quizalofop* A 99.5 g/l EC L/ha (2 3 leaf stage) L/ha (dependent on growth stage and species of weed) Apply specified rate as a post-emergence spray over the top of the pigeon pea crops. (Per13758) Always apply with D-C-trate at 2 L/100 L or Hasten or Kwickin at 1 L/100 L. Uptake at 500 ml/100 L spray volume. The lower doses will provide effective control if applied under ideal conditions to weed that are smaller, actively growing and free from temperature or water stress. (Per13758) Refer to permit for growth stages of species and critical comments. (Per13758) Diquat L 200 g/l AC and SL 2 3 L/ha Harvest aid Diquat/paraquat L 115 g/l g/l SC L/ha Apply pre-sowing, in minimum L water. Apply specified rates for certain weeds at particular growth stages, refer to label. 330 g/l EC L/ha Pendimethalin D 435 g/l EC L/ha 440 g/l EC L/ha 455 g/l EC L/ha Incorporate into the soil within 24 hours of application. Use higher rate on heavy textured soils or those high in organic matter. May be applied by aerial or ground spraying. In Macquarie Valley area, only apply by air when ground is too wet for ground application. 475 g/l EC L/ha Metribuzin C 480 g/l SC 700 g/kg WG L/ha 0.47 kg/ha Furrow irrigated: apply after furrowing out, within 2 weeks before sowing and incorporate. For post-emergence: apply to actively growing seedling stage weeds provided crop plants have at least 2 trifoliate leaves. Do not spray if rain 750 g/kg WG 0.47 kg/ha is likely to fall within several hours. Overhead irrigated: apply pre-emergence then irrigate. *Use of these products is under permit (Per13758). NOTE: Only apply to pigeon pea crops that are to be destroyed at the end of the season or to be harvested for seed for refuge replanting only. No crop product or crop residue is to be fed to livestock. Refer to all labels and permit conditions. Please go to to check allowable usages. Source: Pest Management Guide (pp. 84) Table /17 Pest Management Guide Herbicides available for use in pigeon pea

8 Spray Limitations in the Pigeon Pea Refuge: No Bollgard 3 sprays (Dipel TM ) are allowed for the full season. No insecticides that control Helicoverpa spp. are allowed for the full season. Sunrise New pigeon pea variety for 2017/18 planting An ongoing, commitment towards improving the quality of refuge crops has enabled a new variety of pigeon pea to be developed, which is available to cotton growers this season. Pigeon pea represents about 49 percent of the refuge area cultivated, mainly due to its high productivity as a refuge crop allowing growers to plant a smaller area of refuge, and maximise their commercial cotton production area. Monsanto Entomologist, Dr Kristen Knight, says that whilst pigeon pea is a popular refuge option, it is critical that we continue to research and develop high performing varieties for Australian cotton growers. Cotton growers have found it difficult to reliably source Quality Assured pigeon pea seed over the past 10 years, said Dr Knight. Over this time, the cotton growing landscape as well as grower practices have changed. We have been constantly monitoring refuges to ascertain their attractiveness and effectiveness to ensure the ongoing sustainability of Bt technology. In 2014, the TIMS Committee endorsed a Bollgard 3 Resistance Management Plan (RMP) that included changes to the mandated refuge area for crops. The unsprayed pigeon pea refuge requirement was reduced to 2.5 percent of the area of Bollgard 3 cotton, and pigeon pea was included as an option for dryland Bollgard 3 cotton. This emphasised the need for refuges to be well managed to maximise their effectiveness. One of the focus areas for refuge management is seed quality for pigeon pea. In some circumstances, the mixing of the original varieties and saving of seed from undamaged refuges was resulting in the self-selection of peas that flower late in the season. This has resulted in reduced synchrony with the cotton flowering period. Food sprays cannot be used. Trichogramma or any other biological control agent cannot be released. Secondary pests can be controlled in the refuge as long as the product has no activity on Helicoverpa spp. With the cessation of pigeon pea grain production, and therefore no availability of commercial quantities of pure lines, we needed to start afresh. This meant we had the opportunity to ensure growers had access to a variety of pigeon pea most suitable for use as a refuge. A collaboration between Queensland Department of Agriculture and Fisheries (QDAF), Associated Grains and Monsanto has subsequently led to the development of a new variety, Sunrise. Sunrise exhibits excellent vigour under furrow irrigation on a range of soil types, commences flowering prior to January and has a strongly indeterminate growth habit that allows for repeat flowering, particularly after sustaining insect attack. Assessments of the attractiveness of Sunrise to Helicoverpa and resultant pupae production compared with existing pigeon pea varieties have been very promising. Limited quantities of seed were released last season across the cotton growing regions so growers could see the variety first hand. The commercial release of Sunrise for the 2017/18 season will see Sunrise commercially available to growers. Dr Knight says Monsanto will continue to work in partnership with QDAF to collect additional data for the refuge efficacy of Sunrise. We recognise growers have been continuing to improve refuge management year on year and with the introduction of Sunrise, it will make it easier for growers to manage their pigeon pea refuge crops. It is important that we continue to monitor the development of pigeon pea varieties that will assist growers in producing optimal refuges. The information we gather will be critical for future resistance management planning through Monsanto and the Australian cotton industry. If you have any concerns or questions regarding pigeon pea refuge crops please do not hesitate to contact your Monsanto Regional Business Manager.

9 Sunrise Pigeon Pea GROW A GREAT REFUGE TO SLOW DOWN HELICOVERPA RESISTANCE IN AUSTRALIAN COTTON. Sunrise Key advantages over existing industry saved seed Varietal integrity, reliable growth and flowering. This variety has been purposefully selected for use as a Bollgard 3 refuge based on its growth habit and flowering. It will perform reliably unlike much of the seed currently available that has no known heritage and is increasingly failing to flower and therefore is useless as a refuge. Sunrise will typically flower in early January. Excellent germination, Uniformity of seed quality and reliable supply. This new variety has excellent germination characteristics. Should no longer need to plant large compensatory amounts of seed. Excellent stand ability and vigour on flood irrigation. Whilst Sunrise is still susceptible to water logging it is much better adapted compared to existing material in the market place and exhibits very strong vigour. A vigorous grower with an indeterminate flowering pattern that enables Sunrise Pigeon Pea to produce large numbers of flowers and pods over an extended period, so whilst common peas often flower and stop growing Sunrise will continue to re-flower provided it has adequate soil moisture. From a refuge perspective Sunrise is about twice as effective as the common peas that are available producing high numbers of susceptible Helicoverpa over an extended period. More attractive to Helicoverpa than conventional cotton, hence a more efficient refuge option. Planting rate kg/ha. For more information please contact: Geoff Birch Associated Grain Seed Manager Ph: Mobile: geoff@associatedgrain.com.au Website:

10 Unsprayed Cotton Refuge for Bollgard 3 Cotton Key advantages of unsprayed cotton as a refuge Unsprayed cotton refuge must be planted at 5% of the Bollgard 3 area (refer to Tables 4 & 5.) Planting of unsprayed cotton allows for the refuge to be harvested and income also generated from this area. However, the refuge must not be sprayed to control Helicoverpa spp. at any time. Use of Roundup Ready Flex cotton as a refuge has the added benefit of being able to control in crop weeds with Roundup Ready Herbicide with Plant Shield by Monsanto. Cotton refuges act as a host for other Lepidopteran species such as Rough Bollworm. Unsprayed Cotton Bollgard 3 Resistance Management Plan Requirements Unsprayed cotton refuge area requirements Crop Condition % of Bollgard 3 Central Queensland If planted between August 1 October 31 If planted between November 1 December 31 Irrigated & Dryland* Irrigated & Dryland* Unsprayed cotton Unsprayed pigeon peas 5% 10% *Subject to approved management plan by Monsanto Table 4. Central Queensland Bollgard 3 RMP unsprayed cotton requirements

11 Crop Condition % of Bollgard 3 Southern Queensland, NSW and Victoria Irrigated & Dryland Cotton Unsprayed cotton refuge 5% Table 5. Southern Queensland, NSW and Victoria Bollgard 3 RMP unsprayed cotton requirements Control of Volunteer and Ratoon cotton: Volunteer and ratoon cotton, within fields may impose additional selection pressure on Helicoverpa spp. to develop resistance to Bollgard 3 proteins. Ensure the field that the unsprayed cotton is to be sown into is clean of volunteer or ratoon cotton prior to planting. The easiest way to avoid this is to ensure that refuges are not sown into last years Bollgard 3 stubble. Unsprayed cotton refuges must be kept clean of volunteer and ratoon cotton for the entire growing period. For further information on volunteer cotton and ratoon management visit: products/documents/biotech%20topic%20-%20 Volunteer%20and%20ratoon%20control.pdf Unsprayed Cotton defoliation and harvest: Defoliation of the unsprayed cotton refuge cannot commence until defoliation of the Bollgard 3 cotton has commenced. Pupae Destruction in Unsprayed Cotton Refuges: To ensure maximum emergence of late pupae from the cotton refuge, soil disturbance in the refuge should not occur until after the pupae busting in the Bollgard 3 cotton is complete. Ideally, the unsprayed cotton refuge should be left uncultivated until the following spring if possible. Remember, the more susceptible moths that emerge from the refuge the greater the genetic dilution of any resistant survivors. Spray Limitations in the Unsprayed Cotton Refuge: No Bollgard 3 sprays (DipelTM) are allowed for the full season. No Helicoverpa insecticides are allowed for the full season. Food sprays cannot be used. Trichogramma or any other biological control agent cannot be released.

12 Unsprayed Cotton Agronomic Considerations Simple Steps to creating an effective Unsprayed Cotton Refuge: Field Selection: Avoid planting cotton into last year s cotton field. Do not plant unsprayed cotton refuge into cotton stubble that has not been root cut effectively. Planting Seed Quality: Always perform a germination test (utilise CSD s services) prior to sowing if using old seed. Adjust the sowing rate to ensure a plant stand equivalent to the Bollgard 3 cotton is achieved. Weed management: Apply a residual herbicide for long term weed control. Consult the Roundup Ready Flex Cotton Weed Management Guide for clear recommendations for weed control practices in a Roundup Ready Flex cotton or refuge crop. The guide includes a range of herbicides which offer different modes of action throughout the season, reducing the risk of glyphosate resistance developing on your farm and saving you time and money in the future. The Roundup Ready Flex Cotton Weed Resistance Management Plan details strategies that can be implemented to minimise the risk of glyphosate resistance developing in weeds on-farm. These guides are available from: products/pages/cotton-stewardship.aspx Unsprayed Cotton Refuge for Dryland Cotton: The field history of the refuge and the Bollgard 3 cotton fields must have a similar field history. This is to ensure similar stored moisture levels are available for both crops. Choose a conventional Ready Flex cotton variety as the refuge, to improve your weed control options. Remember don t let this year s refuge generate next year s weeds.

13 FOR ADDITIONAL INFORMATION PLEASE CONTACT: Rohan Sippel Regional Business Manager Central Queensland & Dawson/Callide Mark Dawson Sales Lead Keryn McLean Marketing Lead Kate Lang Product and Stewardship Manager Mick Fing Regional Business Manager Darling Downs & St George/Dirranbandi Tom Luff Regional Business Manager Gwydir, Macintyre & Mungindi Paul Brady Regional Business Manager Namoi & Walgett Luke Sampson Regional Business Manager Lachlan, Macquarie, Bourke & Southern NSW Dr Kristen Knight Entomologist Monsanto Australia Head Office: Level 12, 600 St Kilda Road, Melbourne, VIC 3004 Post: PO Box 6051, St Kilda Road Central, VIC 8008 Phone: Fax: Roundup Ready Flex, Bollgard II and Bollgard 3 are registered trademarks of Monsanto Technologies LLC, used under licence by Monsanto Australia Ltd. Cotton Choices is a trademark of Monsanto Technology LLC used under license by Monsanto Australia Ltd. All other marks are the property of their respective owners. Insect control technology incorporated into these seeds is commercialised under a license from Syngenta Crop Protection AG. MON1422