Overcoming barriers to the successful implementation of a classical biological control strategy for the exotic invasive weed Mikania micrantha in the Asia-Pacific region

Mikania micrantha (Asteraceae) commonly known as mikania, is a major invasive alien plant (IAP) in the tropical humid agricultural and forest zones of the Asia-Pacific region. This fast-growing Neotropical vine is able to smother plants in agricultural ecosystems, agroforestry and natural habitats, reducing productivity and biodiversity. Fungal pathogens were first investigated for the classical biological control of this weed in 1996. This resulted in the selection and screening of the highly host-specific and damaging rust pathogen, Puccinia spegazzinii (Pucciniales). It was first released in India and China in 2005/6, although it is not believed to have established. Since then, it has been released successfully in Taiwan, Papua New Guinea (PNG), Fiji and most recently Vanuatu. The rust has established and is spreading rapidly after applying lessons learned from the first releases on the best rust pathotype and release strategy. In PNG, direct monitoring of vegetation change has demonstrated that the rust is having a significant impact on M. micrantha, with no unpredicted non-target impacts. Despite this, the authorities in many countries where mikania is a problem remain cautious about releasing the rust. In Western Samoa, introduction of the rust was not pursued because of a conflict of interest, and the perception that mikania suppresses even worse weeds. For some, ‘pathophobia’ is still a major obstacle. In Indonesia, where insects for weed CBC have been introduced, pathogens will currently not be considered. In other countries such as Bhutan and Myanmar, there are no baseline data on the presence and impact of IAPs and, with no history of CBC, no institutional framework for implementing this approach. Malaysia has a well-developed framework, but capacity needs to be built in the country. Overall, it remains critical to have champions at decision making levels. Hence, even with an effective ‘off-the-shelf’ agent available, implementation of mikania CBC still requires significant inputs tailored to the countries’ specific needs.

Biological control of bellyache bush (Jatropha gossypiifolia) in Australia: South America as a possible source of natural enemies

Bellyache bush (Jatropha gossypiifolia, Euphorbiaceae), a deciduous shrub introduced as an ornamental from tropical America, is a major and expanding weed of rangelands and riparian zones in northern Australia. Biological control is the most economically viable and long-term management solution for this weed. Surveys for potential biological control agents for J gossypiifolia in Mexico,Central America and the Caribbean resulted in release of the seed-feeding jewel bug Agonosoma trilineatum (Hemiptera: Scutelleridae), which failed to establish, and prioritisation of a leaf-rust Phakopsora arthuriana (Puccineales: Phakopsoraceae) for host-specificity testing, which is ongoing. With poor prospects for new agents from Mexico and Central America and the Caribbean, the search for candidate agents on J gossypiifolia shifted to localities south of the equator. Surveys were conducted on the purple-leaf form of J gossypiifolia, Jatropha excisa, Jatropha clavuligera and Jatropha curcas in Peru, Bolivia and Paraguay in 2012 and 2013. A total of 11 insect species, one mite species and the leaf-rust (P. arthuriana) were observed. These include a yet to be described leafmining moth (Stomphastis sp.) (Lepidoptera: Gracillaridae), a shoot and leaf-galling midge Prodiplosis longifila, and leaf-feeding midge Prodiplosis sp. near longifila (both Diptera:Cecidomyiidae) and an unidentified leaf-feeding moth larva (Lepidoptera: Notodontidae). The leafminer is widespread and damaging and has a field host range restricted to the genus Jatropha in Peru and Bolivia, holds the greatest promise as a biological control agent in Australia. Phakopsora arthuriana was recorded for the first time ever from Bolivia and Peru. Further exploration will be conducted in Peru and Bolivia during the wet season to confirm the field host range of collected agents,and to look for more new agents. Promising agents with field host-range restricted to Jatropha spp. will be imported into a quarantine facility in Australia for host-specificity testing.

Integrating different approaches in the definition of biological stocks: A northern Australian multi-jurisdictional fisheries example using grey mackerel, Scomberomorus semifasciatus

A holistic approach to stock structure studies utilises multiple different techniques on the same individuals sampled from selected populations and combines results across spatial and temporal scales to produce a weight of evidence conclusion. It is the most powerful and reliable source of information to use in formulating resource management and monitoring plans. Few examples of the use of a holistic approach in stock structure studies exist, although more recently this is changing. Using such an approach makes integration of results from each technique challenging. An integrated stock definition (ISD) approach for holistic stock structure studies was developed in this study to aid in the appropriate interpretation of stock structure results to guide the determination of fishery management units. The ISD approach is applied herein to a study of the northern Australian endemic grey mackerel, Scomberomorus semifasciatus (Scombridae). Analyses of genetic (mitochondrial DNA and microsatellites), parasite, otolith stable isotope, and growth data are synthesised to determine the stock structure of S. semifasciatus across northern Australia. Integration of the results from all techniques identified at least six S. semifasciatus stocks for management purposes. Further, the use of the ISD approach provided a simple basis for integrating multiple techniques and for their interpretation. The use of this holistic approach was a powerful tool in providing greater certainty about the appropriate management units for S. semifasciatus. Future stock structure studies investigating spatial management questions in the fisheries context should adopt a holistic approach and apply the ISD approach for a more accurate definition of biological stocks to improve fisheries management.

Life cycle and host range of Phycitasp. rejected for biological control of prickly acacia in Australia

Prickly acacia (Vachellia nilotica subsp. indica), a native of the Indian subcontinent, is a serious weed of the grazing areas of northern Australia and is a target for classical biological control. Native range surveys in India identified a leaf webber, Phycita sp. (Lepidoptera: Pyralidae) as a prospective biological control agent for prickly acacia. In this study, we report the life cycle and host-specificity test results Phycita sp. and highlight the contradictory results between the no-choice tests in India and Australia and the field host range in India. In no-choice tests in India and Australia, Phycita sp. completed development on two of 11 and 16 of 27 non-target test plant species, respectively. Although Phycita sp. fed and completed development on two non-target test plant species (Vachellia planifrons and V. leucophloea) in no-choice tests in India, there was no evidence of the insect on the two non-target test plant species in the field. Our contention is that oviposition behaviour could be the key mechanism in host selection of Phycita sp., resulting in its incidence only on prickly acacia in India. This is supported by paired oviposition choice tests involving three test plant species (Acacia baileyana, A. mearnsii and A. deanei) in quarantine in Australia, where eggs were laid only on prickly acacia. However, in paired oviposition choice trials, only few eggs were laid, making the results unreliable. Although oviposition choice tests suggest that prickly acacia is the most preferred and natural host, difficulties in conducting choice oviposition tests with fully grown trees under quarantine conditions in Australia and the logistic difficulties of conducting open-field tests with fully grown native Australian plants in India have led to rejection of Phycita sp. as a potential biological control agent for prickly acacia in Australia.

The leaf-feeding geometrid Isturgia disputaria (Guenee)-A potential biological control agent for prickly acacia, Vachellia nilotica subsp. indica (Benth.) Kyal. & Boatwr. (Mimosaceae) in Australia

Prickly acacia (Vachellia nilotica subsp. indica), a native multipurpose tree in India, is a weed of National significance, and a target for biological control in Australia. Based on plant genetic and climatic similarities, native range surveys for identifying potential biological control agents for prickly acacia were conducted in India during 2008-2011. In the survey leaf-feeding geometrid, Isturgia disputaria Guenee (syn. Tephrina pulinda), widespread in Tamil Nadu and Karnataka States, was prioritized as a potential biological control agent based on field host range, damage potential and no choice test on non target plant species. Though the field host range study exhibited that V. nilotica ssp. indica and V. nilotica ssp. tomentosa were the primary hosts for successful development of the insect, I. disputaria, replicated no - choice larval feeding and development tests conducted on cut foliage and live plants of nine non-target acacia test plant species in India revealed the larval feeding and development on three of the nine non-target acacia species, V. tortilis, V. planiferons and V. leucophloea in addition to the V. nilotica ssp. indica and V. nilotica ssp. tomentosa. However, the proportion of larvae developing into adults was higher on V. nilotica subsp. indica and V. nilotica subsp. tomentosa, with 90% and 80% of the larvae completing development, respectively. In contrast, the larval mortality was higher on V. tortilis (70%), V. leucophloea (90%) and V. planiferons (70%). The no-choice test results support the earlier host specificity test results of I. disputaria from Pakistan, Kenya and under quarantine in Australia. Contrasting results between field host range and host use pattern under no-choice conditions are discussed.

The host range of three biotypes of Dactylopius tomentosus (Lamarck) (Hemiptera: Dactylopiidae) and their potential as biological control agents of Cylindropuntia spp. (Cactaceae) in Australia

The host range of two newly imported biotypes of Dactylopius tomentosus and their potential as biological control agents of Cylindropuntia spp. were investigated. A third biotype (imbricata) of D. tomentosus previously released in Australia to control C. imbricata was also screened to determine if it will feed on other species of Cylindropuntia occurring in Australia. Efficacy trials were conducted to evaluate the ability of the biotypes to retard the growth or kill those plant species supporting development of four or more individuals in the host test trials. The host range of the three biotypes of D. tomentosus was restricted to the genus Cylindropuntia. However, the biotypes showed varying degrees of specificity within this genus. The imbricata biotype was the only biotype to develop on Australian C. rosea provenances, albeit with a range of developmental success on all C. rosea provenances tested. The Spanish provenance supported the highest development success followed by Grawin (NSW), Lorne Station (NSW) while the least preferred was the Mexican provenance. The rosea and cholla biotypes were unsuitable candidates to control C. rosea in Australia. However, the efficacy trials showed that the cholla biotype had a high impact on four of the eight naturalised Cylindropuntia species in Australia. This biotype established rapidly and the sustained feeding of one fecund female and her progeny killed potted plants of C. imbricata and C. fulgida at week 18. This biotype has the potential to be an effective agent against C. fulgida, C. imbricata, C. kleiniae and C. tunicata and, as a consequence, an application seeking its release in Australia has been lodged.

The combined effect of biological control with plant competition on the management of parthenium weed (Parthenium hysterophorus l.)

Parthenium hysterophorus L., (Asteraceae) commonly known as parthenium weed, is a highly invasive plant that has become a problematic weed of pasture lands in Australia and many other countries around the world. For the management of this weed, an integrated approach comprising biological control and plant competition strategies was tested in southern central Queensland. Two competitive pasture plant species (butterfly pea and buffel grass), selected for their high competitive ability, worked successfully with the biological control agent (Epiblema strenuana Walker) to synergistically reduce the biomass of parthenium weed, by between 62 and 69%. In the presence of biological control agent, the corresponding biomass of competitive plants, butterfly pea and buffel grass increased in comparison to when the biological control agent had been excluded, by 15 and 35%, respectively. This suggests that biological control and competitive plants can complement one another to bring about improved management of parthenium weed in Australia. Further, this approach may be adopted in countries where some of the biological control agents are already present including South Africa, Ethiopia, India, Pakistan and Nepal.

Biological data and model development for management of longfinned eels

Objectives: 1. Estimate population parameters required for a management model. These include survival, density, age structure, growth, age and size at maturity and at recruitment to the adult eel fishery. Estimate their variability among individuals in a range of habitats. 2. Develop a management population dynamics model and use it to investigate management options. 3. Establish baseline data and sustainability indicators for long-term monitoring. 4. Assess the applicability of the above techniques to other eel fisheries in Australia, in collaboration with NSW. Distribute developed tools via the Australia and New Zealand Eel Reference Group.

The host specificity and climatic suitability of the gall flyCecidochares connexa(Diptera: Tephritidae), a potential biological control agent for Chromolaena odorata(Asteraceae) in Australia

The gall fly Cecidochares connexa (Diptera: Tephritidae) is a potential biological control agent for Chromolaena odorata in Australia. Its host specificity was determined against 18 species in the tribe Eupatorieae (Family Asteraceae) in which C. odorata belongs, in quarantine in Brisbane, Australia. Oviposition occurred and flies developed on only C. odorata and Praxelis clematidea, both of which are in the subtribe Praxelinae. P. clematidea is considered a weed outside tropical America. In both multiple-species-minus-C. odorata choice tests and single-species no-choice tests, the mean number of galls/plant was significantly greater on C. odorata (48 and 41, respectively) than on P. clematidea (2 and 9, respectively). There were also significantly more adults emerging from C. odorata (mean 129 and 169, respectively) in the two types of tests than from P. clematidea (1 and 8, respectively). Paired choice, multiple generation (continuation) and time dependent tests further clarified the extent that C. connexa could develop on P. clematidea. In these tests, the mean number of galls formed and the mean number of emerging adults were consistently less for P. clematidea than C. odorata and populations of C. connexa could not be maintained on P. clematidea. Galls were not seen on any other plant species tested. This study supports the results of host specificity testing conducted in seven other countries and confirms that C. connexa poses little risk to other plant species in Australia. C. connexa has been released in 10 countries and an application seeking approval to release in Australia has been submitted to the Australian Government.

The host range of four new biotypes of Dactylopius tomentosus (Hemiptera: Dactylopiidae) from southern USA and their potential as biological control agents of Cylindropuntiaspp.(Cactaceae) in Australia: Part II.

Eight Cylindropuntia species have naturalised in Australia and pose serious economic, environmental and social impacts. Two biotypes of Dactylopius tomentosus have been used as bio-control agents to control different Cylindropuntia species. The host range of four additional biotypes of Dactylopius tomentosus from southern USA was investigated. Feeding and development were restricted to the genus Cylindropuntia. However, they showed differences in specificity within this genus and some biotypes discriminated between the provenances of C. rosea and C. tunicata. Efficacy trials were conducted to determine whether populations of each biotype could be sustained on the naturalised Cylindropuntia species and if these populations could retard the growth or kill these plants. The acanthocarpa biotype offers potential control of C. rosea (Lorne Station), while the cylindropuntia sp. biotype shows great potential to control C. rosea (Grawin). The cylindropuntia sp. biotype also had a high impact on C. kleiniae and C. imbricata, and a moderate impact on C. leptocaulis and C. prolifera. The acanthocarpa X echinocarpa biotype had its greatest impact on C. tunicata (Grawin), killing this plant in 18 weeks. A fourth biotype, leptocaulis, was damaging to some species, but was less effective than the other biotypes. Cylindropuntia spinosior is the only naturalised species in Australia where no effective biocontrol agent has been found.

Bending the Rules of Cell Protrusions : Molecular Mechanisms and Biological Roles of Inverse-BAR Proteins in Cell Morphogenesis

Plasma membrane adopts myriad of different shapes to carry out essential cellular processes such as nutrient uptake, immunological defence mechanisms and cell migration. Therefore, the details how different plasma membrane structures are made and remodelled are of the upmost importance. Bending of plasma membrane into different shapes requires substantial amount of force, which can be provided by the actin cytoskeleton, however, the molecules that regulate the interplay between the actin cytoskeleton and plasma membrane have remained elusive. Recent findings have placed new types of effectors at sites of plasma membrane remodelling, including BAR proteins, which can directly bind and deform plasma membrane into different shapes. In addition to their membrane-bending abilities, BAR proteins also harbor protein domains that intimately link them to the actin cytoskeleton. The ancient BAR domain fold has evolved into at least three structurally and functionally different sub-groups: the BAR, F-BAR and I-BAR domains. This thesis work describes the discovery and functional characterization of the Inverse-BAR domains (I-BARs). Using synthetic model membranes, we have shown that I-BAR domains bind and deform membranes into tubular structures through a binding-surface composed of positively charged amino acids. Importantly, the membrane-binding surface of I-BAR domains displays an inverse geometry to that of the BAR and F-BAR domains, and these structural differences explain why I-BAR domains induce cell protrusions whereas BAR and most F-BAR domains induce cell invaginations. In addition, our results indicate that the binding of I-BAR domains to membranes can alter the spatial organization of phosphoinositides within membranes. Intriguingly, we also found that some I-BAR domains can insert helical motifs into the membrane bilayer, which has important consequences for their membrane binding/bending functions. In mammals there are five I-BAR domain containing proteins. Cell biological studies on ABBA revealed that it is highly expressed in radial glial cells during the development of the central nervous system and plays an important role in the extension process of radial glia-like C6R cells by regulating lamellipodial dynamics through its I-BAR domain. To reveal the role of these proteins in the context of animals, we analyzed MIM knockout mice and found that MIM is required for proper renal functions in adult mice. MIM deficient mice displayed a severe urine concentration defect due to defective intercellular junctions of the kidney epithelia. Consistently, MIM localized to adherens junctions in cultured kidney epithelial cells, where it promoted actin assembly through its I-BAR andWH2 domains. In summary, this thesis describes the mechanism how I-BAR proteins deform membranes and provides information about the biological role of these proteins, which to our knowledge are the first proteins that have been shown to directly deform plasma membrane to make cell protrusions.