This workshop will be taught by seven international Fusarium experts. Participants will be introduced to standard morphological, genetic and molecular biological techniques used to identify and characterize strains of Fusarium. Participants will learn to use morphological characters to identify the most common Fusarium species, how to make tests for vegetative compatibility groups (VCGs) and cross-fertility, and how to extract, PCR amplify DNA, and to analyze DNA sequences. More than half of the time will be spent in the laboratory working with standard strains. Students may bring some of their own strains.

Topics include:

Laboratory Strain Identification

VCG Analysis

Mating Types and Crosses

Species Concepts

Molecular Identification

Mycotoxigenic species

Attendance will also broaden my Fusarium expertise as well as increase my international contacts with Fusarium experts.

The three major areas of learning will involve:

1.) Knowledge and technical expertise to correctly identify numerous Fusarium species using morphological techniques as well as molecular sequencing.

2). Learn traditional techniques including the preparation of single spore cultures, vegetative compatibility grouping, and extensive familiarisation with the morphological characteristic of a range of Fusaria grown on specialised media.

3). Learn molecular identification of Fusarium species including how to extract DNA from the isolate, amplification of DNA using PCR, DNA sequence, and running the DNA sequence through a sequence database.

The Cotton Australia Grower RD&E Advisory Panels provide a critical role within the cotton industry by providing practical advice on research, development and extension needs and priorities. This advice is important guidance to CRDC in its formation of five-year Strategic R&D Plans, Annual Operational Plans, Expressions of Interest for RD&E and resultant CRDC decisions as to project investments.

Cotton Australia facilitates 4 advisory panels that are aligned with the CRDC strategic plan priorities. The panels consist of up to 40 grower, consultant and ginners members from every cotton growing region.

The TIMS committee is facilitated by Cotton Australia and CRDC. It functions as a cotton industry stewardship group, with broad representation from growers, research organisations, crop consultants and members of the pulse and grains industries. The agricultural chemical, biotechnology and planting seed companies that provide crop protection tools to Australian cotton growers approach the TIMS Committee for advice on issues associated with developing or amending resistance management plans for new or existing technologies. Cotton Australia is represented by 6 grower representatives, the TIMS Committee Chair, the Chairs of the three technical panels and the Executive Officer.

The Cotton Australia-facilitated Industry Biosecurity Group (IBG) has traditionally met annually to ensure that the cotton industry’s responsibilities under the Emergency Plant Pest Response (EPPR) Deed are met. Development of an implementation plan through recent revision of the Industry Biosecurity Plan has led to a need to formalise this group to address key biosecurity needs of the industry. It is anticipated that the formalised IBG will constitute 1 staff and up to 3 grower representatives of Cotton Australia as well as representatives from CRDC, Crop Consultants Australia, CSIRO, NSW DPI, QDAF and Plant Health Australia.

The Green Vegetable Bug, Nezara viridula, has recently become a more significant pest of Australian cotton but it is not a problem every season. This project addressed aspects of the multiple host use and movement of N. viridula as they relate to cotton, as well as the genetic relationship between Australian N. viridula and the global populations of this pest.

Samples of N. viridula were collected from northern and eastern Australia and from a variety of weed and crop hosts, with an emphasis on cotton. The abundance of N. viridula was low for the duration of the project but about 800 adult insects were collected overall. Comprehensive phylogenetic and population genetics methods were used to address each of the questions. The methods developed during this study will be made available to other researchers through scientific publications.

The Australia populations of N. viridula come from two different evolutionary lineages, one European and one Asian. The former is distributed across eastern Australia and the other across northern Australia. At some point in the past some individuals of the Asian lineage have mated with individuals of the European lineage in northern Queensland but these events appear to have occurred only rarely. Across the different host plant species there are no genetic differences between N.viridula that would indicate separate host-specific gene pools.

The N. viridula in eastern Australia are more genetically distinct from one another the greater the geographic distance that separates the sampling localities from which they were collected. This slight genetic differentiation over geographic distance is present in insects collected across two years and this indicates that N. viridula populations remain relatively localised in the short term. This result indicates that the host plants available to N. viridula within each cotton growing region will be the most relevant for predicting the abundance of this insect in cotton. Pest pressure from N. viridula was low for the duration of the project and so this pattern may be different during seasons when N. viridula is present in high numbers. In years of high abundance host plants might be found between growing regions, and allow for the recruitment of N. viridula over a wider area.

Future research that addresses the host use of N. viridula should investigate populations from each cotton growing region independently, as local conditions, such as the crop and weed host plants used by N. viridula each season before cotton becomes attractive, will be the most relevant to late season numbers of this insect in cotton. A previous CRDC funded project has already addressed N. viridula host use in central New South Wales. If cotton is grown regularly in northern Australia then it would be prudent to treat the N. viridula population there as a separate entity, as there may be significant differences in their biology which could affect their host use and abundance in cotton. Any differences would therefore influence the development of management strategies.

Cotton leaf curl disease (CLCuD) presents a major biosecurity threat to the Australian industry. The exotic Cotton blue disease (CBD) is also of concern. The industry has invested in preparedness for CLCuD and CBD through project research which commenced in 2008. These projects delivered surveillance of Australian cotton crops, a draft contingency plan for each exotic disease and a surveillance strategy for detection of CLCuD through trapping its whitefly vector.

The 2012 workshop on whitefly transmitted viruses highlighted a need for ongoing investment in this area and a need for improved testing of imported plant material. The detection of Bemisia tabaci Biotype B in northern Australia is a concern given the close proximity with Indonesia. Results indicated Indonesia has many different begomoviruses, although the full diversity is unknown. The diversity of these viruses and satellites in other areas north of Australia is also unknown. The detection of an exotic aphid-transmitted virus of vegetables in Kununurra and Darwin in 2011 demonstrates a potential pathway into northern Australia. A structured surveillance system for exotic cotton viruses both pre- and post-border was the major aim of this project.

The purpose of this travel to the USA was to visit the US Department of Agriculture Arid-land Research Center during 16 May – 31 May 2014. Supervised by Dr. James Hagler, to learn the protocols for Enzyme-Linked Immunosorbent Assay (ELISA), and how to use this technique to identify the presence of pests in the gut of natural predators in cotton fields.

ELISA is a very sensitive test that detects the presence of a specific protein using antibody tagging. These specific proteins can be used to mark prey items (pests), which can then be identified in the gut-content of a predator (beneficial in cotton fields). We evaluated two methods: indirect ELISA and “sandwich” ELISA; and two protein marks: rabbit and chicken IgG. There was a difference in the detectability of the protein tags in different pest preys. Whitefly marks were difficult to detect, regardless of the method of application, while small topical marks on the tarnished bug Lygus spp showed positive in the assays.

The CRDC funded “Automation of recycle system- IREC Field Station” project saw the supply & installation of variable speed drive pump, sensor and recycle pump automation technology to complement the state-of-the-art automated irrigation system in use.

This project enables IREC to demonstrate to irrigators and visitors the benefits associated with a fully integrated automated irrigation and recycle system.

It has been funded significantly by CRDC with contributions from MI, Bidgee Automation, Padman Stops and other commercial businesses.

The IREC Field Station will now be able to exhibit a wide range of technologies and, management practices as well as share knowledge gained from the various research projects taking place at the site.

Bt cotton has enabled the cotton industry to reduce its use of insecticides and improve the lifestyles of cotton growers by producing a more reliable and sustainable crop through controlling the major pests, Helicoverpa armigera and H.punctigera. However, unexpectedly high numbers of H. armigera and H. punctigera moths have been emerging from Bt cotton field tests, suggesting that about half the moths emerging from the cotton/refuge system could be originating from Bt cotton.

As the moths emerging from refuges do not appear to be overwhelming those emerging from Bt cotton, the above results indicate a potential weakness in the refuge strategy of the Resistance Management Plan (RMP). Consequently, we need to quantify the relative number of moth emergences from Bt cotton and refuges as Bollgard III is adopted, to test that our anti-resistance strategies are working with Bollgard III.

Given the apparently low numbers of moths emerging from refuges, other sources of susceptible moths, such as unstructured refuges, may be more important than previously expected. Researchers will measure the ability of some unstructured refuges to produce moths and therefore support the RMP.

From the RMP perspective, survival in Bollgard III is relative to survival in refuges, but refuge attractiveness & productivity is very variable, again compromising the refuge strategy. Here we aim to reduce refuge variability by calibrating moth emergence from well managed and poorly managed refuges.

Despite the higher than expected numbers of Helicoverpa emerging from Bt cotton, these moths were not resistant to Bt toxins. Instead, they apparently survived on poorly expressing flowers and bolls and therefore could have been tolerating low amounts of toxin. Work from CES 1304 indicated that the F2 generation of field caught moths tolerated proportionately higher levels of Cry1Ac toxin (c. 8x higher) than Cry2Ab toxin (NS) when compared to laboratory colonies. There was further evidence that moths emerging from Bt crops were particularly tolerant to Cry1Ac.

Although Bollgard III with Vip3A will reduce the resistance risk, both the expression of Vip3A and the resistance frequencies of Helicoverpa are less than ideal, so control via Cry1Ac & Cry2Ab remain crucial. Consequently, researchers will continue the work of CES 1304 to compare the level of resistance and amount of tolerance to Bt toxins in moths emerging from Bt crops and refuges.

Travel to the USDA Kika De La Garcia Agricultural Research Centre at Weslaco in South

West Texas. This is an area of mixed cropping that includes cotton, sugarcane, soybean, corn,

and a range of other crops. Dr John Adamczyk, who is based at this institute, visited Narrabri

in 2007 to review research on IPM and develop collaborations (costs were largely covered by

USDA). Dr Adamczyk has extensive experience, especially with the implementation of Bt cottons

and evaluation of factors affecting their field performance and efficacy. He also has

considerable experience with management and research on other cotton pests. This visit was

valuable, especially in planning some of our own research to understand the interaction

between mirid control and mite outbreaks. The discussions also significantly benefitted

Cotton CRC PhD student Baoqain Lu in planning his research to understand how susceptible

Helicoverpa larvae survive in cotton and the economic significance of the damage they

cause.

Dr Adamczyk invited me to visit in June to review cotton research they are doing with the

natural enemy complex in the cotton/soybean system. However, I spent most of my time on

other issues including reviewing progress with Bao’s thesis, mostly directly related to cotton

production, but some related to management of other pests in different systems – which I

thought may be a source of ideas that could be relevant to cotton. I was asked to present a

seminar on cotton research in Australia, especially in relation to management of mirids in

Bollgard II cotton systems.

The Cotton export market is highly competitive and when it comes to quality Australia needs to be

the world's best. To realise this goal, the whole of the Australian Cotton supply chain must

continuously improve its supply of premium upland cotton.

Cotton spinning mills already recognise that Australian cotton has desirable fibre characteristics and

low contamination. These attributes increase efficiency for spinners and they actively seek

Australian cotton and are sometimes prepared to pay a premium. To maintain this reputation

continuous improvement across the whole supply chain is essential.

The Australian cotton industry and CSIRO have expanded investment in post-harvest cotton

processing research. The aim is to discover ways of maintaining and enhancing the quality of cotton

produced by Australian growers.

In July 2008 Rene van der SIuijs and the CSIRO team in Geelong opened the doors of their facility and

hosted the 7th 'Cotton Field to Fabric Course'. This was the 7th course run in Geelong and it has been

attended by participants from the length and breadth of the supply chain. They have included

Agronomists, Growers, Researchers, Ginners and even students studying design.

The course provided participants with an opportunity to see firsthand how cotton is processed from

a bale into fabric. At Geelong they have both full scale and miniature versions of the equipment used

in cotton processing factories used overseas including drawing and carding machines, spinning

frame, weaving machines, and dyeing facilities. Understanding how these processes occur helps

participants understand the importance quality standards and how our actions impact on the chain.

The Australian cotton industry will benefit from a focus on its customer's needs and a desire to

exceed their expectations. The' field to fabric 'course is one activity that the industry is undertaking

to increase knowledge of cotton quality. It comes highly recommended by all who have

participated.

The major output from this trip was to disseminate my latest research results to peers by presenting a spoken paper entitled "Frequency and characteristics of alleles conferring resistance to the Bt toxin Cry2Ab in populations of the low profile target pest Helicoverpa punctigera". In this talk I presented the first information on Cry2Ab resistance in H. punctigera in a context that complimented a prior presentation by Rod Mahon on Cry2Ab resistance in H. armigera. These presentations were attended by leading experts throughout the world in the field of Bt resistance, and were well received. Since returning from South Africa I have written some of this work into a scientific paper that is currently in internal review within CSIRO.

The talks that I attended in the symposia on “Pesticides, Resistance and Transgenics” gave an international overview of the latest thinking in this field. Many of the presentations were review papers rather than disseminations of empirical data. Stimulated largely by a recent controversial paper by Tabashnik et al.2, there was considerable discussion about the definition of resistance and the merits of different approaches for measuring it. A couple of presenters noted that the significant workload involved with performing F2/F1 screens precluded them from being a preferred method, and highlighted the potential danger of focusing on one gene(s) using the F1 tests while neglecting others. However, the feedback from these people after our talks was positive and complimentary of our approach, and acknowledged the appropriateness of the method for our situation.