Reproductive strategies of two invasive tilapia species Oreochromis mossambicus and Tilapia mariae in northern Australia

The reproductive biology of two invasive tilapia species, Oreochromis mossambicus and Tilapia mariae, resident in freshwater habitats in north-eastern Australia was investigated. Oreochromis mossambicus exhibited plasticity in some of its life-history characteristics that enhanced its ability to occupy a range of habitats. These included a shallow, weed-choked, freshwater coastal drain that was subject to temperature and dissolved oxygen extremes and water-level fluctuations to cooler, relatively high-altitude impoundments. Adaptations to harsher conditions included a decreased total length (LT) and age ( A) at 50% maturity (m50), short somatic growth intervals, early maturation and higher relative fecundities. Potential fecundity in both species was relatively low, but parental care ensured high survival rates of both eggs and larvae. No significant difference in the relative fecundity of T. mariae populations in a large impoundment and a coastal river was found, but there were significant differences in relative fecundities between several of the O. mossambicus populations sampled. Total length ( LT) and age at 50% maturity of O. mossambicus populations varied considerably depending on habitat. The LTm50 and Am50 values for male and female O. mossambicus in a large impoundment were considerably greater than for those resident in a small coastal drain. Monthly gonad developmental stages and gonado-somatic indices suggested that in coastal areas, spawning of O. mossambicus and T. mariae occurred throughout most of the year while in cooler, high-altitude impoundments, spawning peaked in the warmer, summer months. The contribution these reproductive characteristics make to the success of both species as colonizers is discussed in the context of future control and management options for tilapia incursions in Australia.

In vitro Induction of Banana Autotetraploids by Colchicine Treatment of Micropropagated Diploids

Alternative breeding strategies, based on colchicine-induced autotetraploids, have been proposed as a means of introducing disease resistance into banana breeding programs. This paper describes techniques for the in vitro induction of banana autotetraploids by the use of colchicine on cultured explants. The technique can be readily applied and large numbers of autotetraploids produced. The optimum treatment involved immersing shoot tips in a 0.5% w/v colchicine solution for 2 h under aseptic conditions. Dimethyl sulfoxide (DMSO) was applied with the colchicine treatments to increase cell permeability and so absorption of colchicine, resulting in the optimum treatment unchanged at 0.5% colchicine, but including the addition of 2% v/v DMSO. Of the shoot tips treated over 30% were induced to the autotetraploid level. Methods for in vitro selection of induced tetraploids from treated diploid plantlets were also developed. Tetraploid plants were more robust with thicker pseudostems, roots and broader leaves than diploids and they could be selected on these morphological characteristics. Mean stornatal lengths of diploid banana plants growing in vitro were significantly smaller (16.0 pm) than the tetraploids (26.9pm) and were used as a more reliable indicator of ploidy than morphological criteria alone. A root tip squash technique using carbol fuchsin was developed for positive confirmation of ploidy change by chromosome counts. Although chimerism and reversion to the diploid form occurred, it was not considered a problem because of the large number of autotetraploids induced. Stable autotetraploids were recovered and established in the field and were characterised by their large, drooping leaves and thick pseudostems. They have retained these characteristics for more than 3 years in the field.

A new data source for fisheries resource assessment: genetic estimates of the effective number of spawners. Final Report to the Fisheries Research and Development Corporation.

The development of innovative methods of stock assessment is a priority for State and Commonwealth fisheries agencies. It is driven by the need to facilitate sustainable exploitation of naturally occurring fisheries resources for the current and future economic, social and environmental well being of Australia. This project was initiated in this context and took advantage of considerable recent achievements in genomics that are shaping our comprehension of the DNA of humans and animals. The basic idea behind this project was that genetic estimates of effective population size, which can be made from empirical measurements of genetic drift, were equivalent to estimates of the successful number of spawners that is an important parameter in process of fisheries stock assessment. The broad objectives of this study were to 1. Critically evaluate a variety of mathematical methods of calculating effective spawner numbers (Ne) by a. conducting comprehensive computer simulations, and by b. analysis of empirical data collected from the Moreton Bay population of tiger prawns (P. esculentus). 2. Lay the groundwork for the application of the technology in the northern prawn fishery (NPF). 3. Produce software for the calculation of Ne, and to make it widely available. The project pulled together a range of mathematical models for estimating current effective population size from diverse sources. Some of them had been recently implemented with the latest statistical methods (eg. Bayesian framework Berthier, Beaumont et al. 2002), while others had lower profiles (eg. Pudovkin, Zaykin et al. 1996; Rousset and Raymond 1995). Computer code and later software with a user-friendly interface (NeEstimator) was produced to implement the methods. This was used as a basis for simulation experiments to evaluate the performance of the methods with an individual-based model of a prawn population. Following the guidelines suggested by computer simulations, the tiger prawn population in Moreton Bay (south-east Queensland) was sampled for genetic analysis with eight microsatellite loci in three successive spring spawning seasons in 2001, 2002 and 2003. As predicted by the simulations, the estimates had non-infinite upper confidence limits, which is a major achievement for the application of the method to a naturally-occurring, short generation, highly fecund invertebrate species. The genetic estimate of the number of successful spawners was around 1000 individuals in two consecutive years. This contrasts with about 500,000 prawns participating in spawning. It is not possible to distinguish successful from non-successful spawners so we suggest a high level of protection for the entire spawning population. We interpret the difference in numbers between successful and non-successful spawners as a large variation in the number of offspring per family that survive – a large number of families have no surviving offspring, while a few have a large number. We explored various ways in which Ne can be useful in fisheries management. It can be a surrogate for spawning population size, assuming the ratio between Ne and spawning population size has been previously calculated for that species. Alternatively, it can be a surrogate for recruitment, again assuming that the ratio between Ne and recruitment has been previously determined. The number of species that can be analysed in this way, however, is likely to be small because of species-specific life history requirements that need to be satisfied for accuracy. The most universal approach would be to integrate Ne with spawning stock-recruitment models, so that these models are more accurate when applied to fisheries populations. A pathway to achieve this was established in this project, which we predict will significantly improve fisheries sustainability in the future. Regardless of the success of integrating Ne into spawning stock-recruitment models, Ne could be used as a fisheries monitoring tool. Declines in spawning stock size or increases in natural or harvest mortality would be reflected by a decline in Ne. This would be good for data-poor fisheries and provides fishery independent information, however, we suggest a species-by-species approach. Some species may be too numerous or experiencing too much migration for the method to work. During the project two important theoretical studies of the simultaneous estimation of effective population size and migration were published (Vitalis and Couvet 2001b; Wang and Whitlock 2003). These methods, combined with collection of preliminary genetic data from the tiger prawn population in southern Gulf of Carpentaria population and a computer simulation study that evaluated the effect of differing reproductive strategies on genetic estimates, suggest that this technology could make an important contribution to the stock assessment process in the northern prawn fishery (NPF). Advances in the genomics world are rapid and already a cheaper, more reliable substitute for microsatellite loci in this technology is available. Digital data from single nucleotide polymorphisms (SNPs) are likely to super cede ‘analogue’ microsatellite data, making it cheaper and easier to apply the method to species with large population sizes.

Host-delivered RNAi: An effective strategy to silence genes in plant parasitic nematodes

Root-knot nematodes (Meloidogyne spp.) are obligate, sedentary endoparasites that infect many plant species causing large economic losses worldwide. Available nematicides are being banned due to their toxicity or ozone-depleting properties and alternative control strategies are urgently required. We have produced transgenic tobacco (Nicotiana tabacum) plants expressing different dsRNA hairpin structures targeting a root-knot nematode (Meloidogyne javanica) putative transcription factor, MjTis11. We provide evidence that MjTis11 was consistently silenced in nematodes feeding on the roots of transgenic plants. The observed silencing was specific for MjTis11, with other sequence-unrelated genes being unaffected in the nematodes. Those transgenic plants able to induce silencing of MjTis11, also showed the presence of small interfering RNAs. Even though down-regulation of MjTis11 did not result in a lethal phenotype, this study demonstrates the feasibility of silencing root-knot nematode genes by expressing dsRNA in the host plant. Host-delivered RNA interference-triggered (HD-RNAi) silencing of parasite genes provides a novel disease resistance strategy with wide biotechnological applications. The potential of HD-RNAi is not restricted to parasitic nematodes but could be adapted to control other plant-feeding pests.

Biological and economic management strategy evaluations of the eastern king prawn fishery

Stock assessment of the eastern king prawn (EKP) fishery, and the subsequent advice to management and industry, could be improved by addressing a number of issues. The recruitment dynamics of EKP in the northern (i.e., North Reef to the Swain Reefs) parts of the fishery need to be clarified. Fishers report that the size of the prawns from these areas when they recruit to the fishing grounds is resulting in suboptimal sizes/ages at first capture, and therefore localised growth overfishing. There is a need to assess alternative harvest strategies of the EKP fishery, via computer simulations, particularly seasonal and monthly or lunar-based closures to identify scenarios that improve the value of the catch, decrease costs and reduce the risk of overfishing, prior to implementing new management measures.

Veneer Recovery Analysis of Plantation Eucalypt Species Using Spindleless Lathe Technology

The Australian hardwood plantation industry is challenged to identify profitable markets for the sale of its wood fibre. The majority of the hardwood plantations already established in Australia have been managed for the production of pulpwood; however, interest exists to identify more profitable and value-added markets. As a consequence of a predominately pulpwood-focused management regime, this plantation resource contains a range of qualities and performance. Identifying alternative processing strategies and products that suit young plantation-grown hardwoods have proved challenging, with low product recoveries and/or unmarketable products as the outcome of many studies. Simple spindleless lathe technology was used to process 918 billets from six commercially important Australian hardwood species. The study has demonstrated that the production of rotary peeled veneer is an effective method for converting plantation hardwood trees. Recovery rates significantly higher than those reported for more traditional processing techniques (e.g., sawmilling) were achieved. Veneer visually graded to industry standards exhibited favourable recoveries suitable for the manufacture of structural products.

Comparison of three pollination methods for Eucalyptus argophloia, a small-flowered eucalypt

Context Most studies assess pollination success at capsule maturity, and studies of pre-zygotic processes are often lacking. Aims This study investigates the suitability of controlled pollination for a potential forestry plantation species, Eucalyptus argophloia, by examining pre- and post-zygotic pollination success. Methods Pollen tube development, capsule set and seed set are compared following three-stop pollination, artificially induced protogyny (AIP), AIP unpollinated and open pollination. The fecundity of stored pollen was compared with that of fresh pollen. Results Three-stop pollination, AIP and open pollination had similar numbers of pollen tubes, but AIP unpollinated had none. Open pollination produced significantly more capsules and total number of seeds than the other treatments. There were significantly more seeds per retained capsule for the open pollination and three-stop pollination treatments than for the AIP and AIP unpollinated pollination treatments. There were no significant differences relative to the age of pollen. Conclusions Pre-zygotic success in terms of pollen tubes was similar for open-pollinated, three stop and AIP, but was not reflected in post-zygotic success when the open pollination and three-stop method produced significantly more seeds per retained capsule than the AIP treatments and open pollination yielded more seeds. Capsule set and total seed set for open pollination, and fewer capsules in controlled pollinations, may reflect physical damage to buds because of the small E. argophloia flowers. Suitable alternative breeding strategies other than controlled pollinations are discussed for this species.

A comparison of alternative plant mixes for conservation bio-control by native beneficial arthropods in vegetable cropping systems in Queensland Australia.

Cucurbit crops host a range of serious sap-sucking insect pests, including silverleaf whitefly (SLW) and aphids, which potentially represent considerable risk to the Australian horticulture industry. These pests are extremely polyphagous with a wide host range. Chemical control is made difficult due to resistance and pollution, and other side-effects are associated with insecticide use. Consequently, there is much interest in maximising the role of biological control in the management of these sap-sucking insect pests. This study aimed to evaluate companion cropping alongside cucurbit crops in a tropical setting as a means to increase the populations of beneficial insects and spiders so as to control the major sap-sucking insect pests. The Population of beneficial and harmful insects, with a focus on SLW and aphids, and other invertebrates were sampled weekly oil four different crops which could be used for habitat manipulation: Goodbug Mix (GBM; a proprietary seed Mixture including self-sowing annual and perennial herbaceous flower species); lablab (Lablab purpureus L. Sweet); lucerne (Medicago sativa L.); and niger (Guizotia abyssinica (L.f.) Cass.). Lablab hosted the highest numbers of beneficial insects (larvae and adults of lacewing (Mallada signata (Schneider)), ladybird beetles (Coccinella transversalis Fabricius) and spiders) while GBM hosted the highest numbers of European bees (Apis mellifera Linnaeus) and spiders. Lucerne and niger showed little promise in hosting beneficial insects, but lucerne hosted significantly more spiders (double the numbers) than niger. Lucerne hosted significantly more of the harmful insect species of aphids (Aphis gossypii (Glover)) and Myzus persicae (Sulzer)) and heliothis (Heliothis armigera Hubner). Niger hosted significantly more vegetable weevils (Listroderes difficillis (Germar)) than the other three species. Therefore, lablab and GBM appear to be viable options to grow within cucurbits or as field boundary crops to attract and increase beneficial insects and spiders for the control of sap-sucking insect pests. Use of these bio-control strategies affords the opportunity to minimise pesticide usage and the risks associated with pollution.

IPM strategies in pome and stonefruit in Western Australia and Queensland

In association with Western Australia develop monitoring and alternative control measures pest thrip species of apples and stonefruit in Queensland.

Developing and communicating strategies for controlling virus diseases in vegetable cucurbit crops

Virus diseases cause serious yield and quality losses in field grown cucurbit crops worldwide. In Australia, the main viruses of cucurbits are Papaya ringspot virus (PRSV), Squash mosaic virus (SqMV), Watermelon mosaic virus (WMV) and Zucchini yellow mosaic virus (ZYMV). Plants infected early have severely distorted fruit. High infection incidences, of ZYMV and PRSV in crops cause losses of marketable fruit of up to 100% and infected crops are often abandoned. Two new alternative hosts of ZYMV were identified, the native cucurbit Cucumis maderaspatanus and wild legume Rhyncosia minima. No new alternative hosts of PRSV, SqMV or WMV were found in Western Australia or Queensland. Seed transmission of ZYMV (0.7%) was found in seedlings grown from ZYMV-infected fruit of zucchini but not of pumpkin. None was detected with PRSV or SqMV in zucchini or pumpkin seedlings, respectively. ZYMV spread to pumpkins by aphids was greater downwind than upwind of a virus source. Delaying sowing by 2 weeks decreased ZYMV spread. Millet non-host barriers between pumpkin plantings slowed ZYMV infection. Host resistance gene (zym) in cucumber cultivars was effective against ZYMV. Pumpkin cultivars with resistance gene (Zym) became infected under high virus pressure but leaf symptoms were milder and infected plants higher yielding with more market-acceptable fruit than those without Zym. Most zucchini cultivars with Zym developed severe leaf and fruit symptoms. ZYMV, PRSV, WMV and SqMV spread readily from infected to healthy cucurbit plants by direct leaf contact. ZYMV survives and remains infective on diverse surfaces for up to 6 hours but can be inactivated by some disinfectants. Phylogenetic analysis indicates at least three separate introductions of ZYMV into Australia, with new introductions rarely occurring. ZYMV isolates clustered into three groups according to collection location i) Kununurra, ii) Northern Territory and iii) Carnarvon, Qld and Vic. A multiplex Real-Time PCR was developed which distinguished between the three groups of Australian isolates. Integrated disease management (IDM) strategies for virus diseases of vegetable cucurbit crops grown in the field were improved incorporating the new information gathered. These strategies are aimed at causing using minimal extra expense, labour demands and disruption to normal practices.

Reproductive variation in naturally occurring populations of the weed Parthenium hysterophorus (Asteraceae) in Australia

Parthenium weed, an annual herb native to tropical America, causes severe economic, human, and animal health and environmental impacts in Australia and in many countries in Asia, Africa, and the Pacific. There is little known about variation in reproductive output in naturally occurring populations of this weed. This information is vital to develop plant population models, devise management strategies to reduce seed output, and formulate parthenium weed pollen-induced human health (e.g., dermatitis and hay fever) risk assessment. Here, the variations in the number of capitula produced by the parthenium weed at two sites in Queensland, Australia, over a 4-yr period are reported. Under field conditions, parthenium weed produced up to 39,192 capitula per plant (> 156,768 seeds per plant), with majority of the plants (approximate to 75%) producing between 11 and 1,000 capitula, and less than 0.3% of the plants producing more than 10,000 capitula (> 40,000 seeds per plant). The number of capitula per plant in the field (297 +/- 22) was much lower than those reported from glasshouse and laboratory studies. Plant biomass contributed to 50 to 80% of the variation in capitulum production between plants within plots at each site, and weed density accounted for 62 to 73% of the variation in capitulum production between plots within each site. As plant size is directly correlated with reproductive output, plant size distributions in parthenium weed can be used to estimate effective population size. Information on variation in reproductive output will be used to implement management strategies to reduce parthenium weed seed output, resulting in reduced soil seed bank and weed seed spread.

Seed bank longevity and age to reproductive maturity of Calotropis procera (Aiton) W.T. Aiton in the dry tropics of northern Queensland

Understanding the reproductive biology of Calotropis procera (Aiton) W.T. Aiton, an invasive weed of northern Australia, is critical for development of effective management strategies. Two experiments are reported on. In Experiment 1 seed longevity of C. procera seeds, exposed to different soil type (clay and river loam), pasture cover (present and absent) and burial depth (0, 2.5, 10 and 20 cm) treatments were examined. In Experiment 2 time to reach reproductive maturity was studied. The latter experiment included its sister species, C. gigantea (L.) W.T. Aiton, for comparison and two separate seed lots were tested in 2009 and 2012 to determine if exposure to different environmental conditions would influence persistence. Both seed lots demonstrated a rapid decline in viability over the first 3 months and declined to zero between 15 and 24 months after burial. In Experiment 1, longevity appeared to be most influenced by rainfall patterns and associated soil moisture, burial depth and soil type, but not the level of pasture cover. Experiment 2 showed that both C. procera and C. gigantea plants could flower once they had reached an average height of 85 cm. However, they differed significantly in terms of basal diameter at first flowering with C. gigantea significantly smaller (31 mm) than C. procera (45 mm). On average, C. gigantea flowered earlier (125 days vs 190 days) and set seed earlier (359 days vs 412 days) than C. procera. These results suggest that, under similar conditions to those that prevailed in the present studies, land managers could potentially achieve effective control of patches of C. procera in 2 years if they are able to kill all original plants and treat seedling regrowth frequently enough to prevent it reaching reproductive maturity. This suggested control strategy is based on the proviso that replenishment of the seed bank is not occurring from external sources (e.g. wind and water dispersal).