Identification of a new Polerovirus (family Luteoviridae) associated with cotton bunchy top disease in Australia

Cotton bunchy top (CBT) disease has caused significant yield losses in Australia and is now managed by control of its vector, the cotton aphid (Aphis gossypii). Its mode of transmission and similarities in symptoms to cotton Blue Disease suggested it may also be caused by a luteovirus or related virus. Degenerate primers to conserved regions of the genomes of the family Luteoviridae were used to amplify viral cDNAs from CBT-affected cotton leaf tissue that were not present in healthy plants. Partial genome sequence of a new virus (Cotton bunchy top virus, CBTV) was obtained spanning part of the RNA-dependent-RNA-polymerase (RdRP), all of the coat protein and part of the aphid-transmission protein. CBTV sequences could be detected in viruliferous aphids able to transmit CBT, but not aphids from non-symptomatic plants, indicating that it is associated with the disease and may be the causal agent. All CBTV open-reading frames had their closest similarity to viruses of the genus Polerovirus. The partial RdRP had 90 % amino acid identity to the RdRP of Cotton leafroll dwarf virus (CLRDV) that causes cotton blue disease, while other parts of the genome were more similar to other poleroviruses. The sequence similarity and genome organization of CBTV suggest that it should be considered a new member of the genus Polerovirus. This partial genome sequence of CBTV opens up the possibility for developing diagnostic tests for detection of the virus in cotton plants, aphids and weeds as well as alternative strategies for engineering CBT resistance in cotton plants through biotechnology. © 2012 Australasian Plant Pathology Society Inc.

Stunted cotton (Gossypium hirsutum L.) fully recovers biomass and yield of seed cotton after delayed root inoculation with spores of an arbuscular mycorrhizal fungus (Glomus mosseae)

In the northern grain and cotton region of Australia, poor crop growth after long periods of fallow, called 'long-fallow' disorder, is caused by a decline of natural arbuscular-mycorrhizal fungi (AMF). When cotton was grown in large pots containing 22 kg of Vertisol from a field recently harvested from cotton in Central Queensland, plants in pasteurised soil were extremely stunted compared with plants in unpasteurised soil. We tested the hypothesis that this extreme stunting was caused by the absence of AMF and examined whether such stunted plants could recover from subsequent treatment with AMF spores and/or P fertiliser. At 42 days after sowing, the healthy cotton growing in unpasteurised soil had 48% of its root-length colonised with AMF, whereas the stunted cotton had none. After inoculation with AMF spores (6 spores/g soil of Glomus mosseae) and/or application of P fertiliser (50 mg P/kg soil) at 45 days after sowing, the stunted plants commenced to improve about 25 days after treatment, and continued until their total dry matter and seed cotton production equalled that of plants growing in unpasteurised soil with natural AMF. In contrast, non-mycorrhizal cotton grown without P fertiliser remained stunted throughout and produced no bolls and only 1% of the biomass of mycorrhizal cotton. Even with the addition of P fertiliser, non-mycorrhizal cotton produced only 64% of the biomass and 58% of the seed cotton (lint + seed) of mycorrhizal cotton plants. These results show that cotton is highly dependent on AMF for P nutrition and growth in Vertisol (even with high rates of P fertiliser), but can recover from complete lack of AMF and consequent stunting during at least the first 45 days of growth when treated with AMF spores and/or P fertiliser. This corroborates field observations in the northern region that cotton may recover from long-fallow disorder caused by low initial levels of AMF propagules in the soil as the AMF colonisation of its roots increases during the growing season.

Reducing the incidence of under-skin browning in 'Honey Gold' mango (Mangifera indica L.) fruit

'Honey Gold' mango is a relatively new cultivar in Australia, with an appealing skin colour and a sweet fibre-free flesh. However, fruit can develop 'under-skin browning' (USB), which appears several days after packing as a distinct 'bruise'-like discolouration under the epidermis and can affect large areas of the fruit surface. We investigated the anatomy of USB and the impact of post-harvest fruit handling conditions on the disorder. Starch accumulated around the resin canals and discoloured cells in the affected area, with no visible change to the cuticle or epidermis. Delays of 1 d at ambient temperature (27 degrees - 35 degrees C) before packing, and 2 d at 18 degrees - 20 degrees C (after packing), before placing fruit at 12 degrees - 14 degrees C and road transportation, reduced the incidence of USB by 83% compared to placing fruit at 12 degrees - 14 degrees C within 13 h of picking. The incidence of USB was 88 100% higher in fruit that were cooled to 12 degrees - 14 degrees C within 13 h of picking, then commercially road-freighted for 4 d at 12 degrees - 14 degrees C, than in fruit held under similar temperature conditions, but not road-freighted. Wrapping each fruit in bubble-wrap to minimise direct contact with other fruit, with the plastic insert, or with the cardboard tray, reduced the incidence of USB by 84% after road-freight compared to not using bubble-wrap. These results suggest that USB is a unique disorder of mango skin associated with a rapid post-harvest reduction in temperature, from high ambient temperatures to 12 degrees - 14 degrees C, and with physical damage during road-freight.

Mango (Mangifera indica L.) peel extract fractions from different cultivars differentially affect lipid accumulation in 3T3-L1 adipocyte cells

Plant phytochemicals are increasingly recognised as sources of bioactive molecules which may have potential benefit in many health conditions. In mangoes, peel extracts from different cultivars exhibit varying effects on adipogenesis in the 3T3-L1 adipocyte cell line. In this study, the effects of preparative HPLC fractions of methanol peel extracts from Irwin, Nam Doc Mai and Kensington Pride mangoes were evaluated. Fraction 1 contained the most hydrophilic components while subsequent fractions contained increasingly more hydrophobic components. High content imaging was used to assess mango peel fraction effects on lipid accumulation, nuclei count and nuclear area in differentiating 3T3-L1 cells. For all three mango cultivars, the more hydrophilic peel fractions 1-3 inhibited lipid accumulation with greater potency than the more hydrophobic peel fractions 4. For all three cultivars, the more lipophilic fraction 4 had concentrations that enhanced lipid accumulation greater than fractions 1-3 as assessed by lipid droplet integrated intensity. The potency of this fraction 4 varied significantly between cultivars. Using mass spectrometry, five long chain free fatty acids were detected in fraction 4; these were not present in any other peel extract fractions. Total levels varied between cultivars, with Irwin fraction 4 containing the highest levels of these free fatty acids. Lipophilic components appear to be responsible for the lipid accumulation promoting effects of some mango extracts and are the likely cause of the diverse effects of peel extracts from different mango cultivars on lipid accumulation.

Pond apple (Annona glabra L.) - Investigating novel mechanical control options

Pond apple usually occurs in swampy areas, but mechanical control may be a viable option in some locations during drier periods. Two machines, the Positrack™ and the Tracksaw™, have been trialled for initial kill rate, amount of follow-up control required, safety to field operators, cost-efficiency and selectivity (effect on native vegetation), compared to other control options. The Positrack™ is a tracked bobcat with a slasher-type attachment that cuts individual trees off near ground level and mulches them. It has no on-board herbicide application capability and requires an additional on-ground operator to apply herbicide by hand. The Tracksaw™ is a tracked mini-excavator with a chainsaw bar and spray applicator on the boom that cuts individual trees off near ground level and applies chemical immediately to the cut stump, requiring only a single operator. Initial trials were done in infestations of similar sizes and densities at the Daintree (Positrack™) and in Innisfail (Tracksaw™) in late 2009. Kill rates to date are 83% for Positrack™ mechanical, 95% for Positrack™ mechanical plus herbicide, and 78% for the Tracksaw™ combined treatment. If ongoing comparison proves either of these machines to be more cost effective, selective, and safer than traditional control methods, mechanical control methods may become more widely used.

Can competition with pasture be used to manipulate bellyache bush (Jatropha gossypiifolia L.) population biology?

Bellyache bush (Jatropha gossypiifolia L.) is an invasive weed that poses economic and environmental problems in northern Australia. Competition between pasture and bellyache bush was examined in North Queensland using combinations of five pasture treatments (uncut (control); cut as low, medium, and high pasture; and no pasture) and four bellyache bush densities (0, 2, 6 and 12plantsm(-2)) in a buffel grass (Cenchrus ciliaris L.) dominated pasture. The pasture treatments were applied approximately once per year but no treatments were applied directly to the bellyache bush plants. Measurements of bellyache bush flowering, seed formation, and mortality were undertaken over a 9-year period, along with monitoring the pasture basal cover and plant species diversity. Maximum flowering rates of bellyache bush occurred after 9 years (97%) in plots containing no pasture, with the lowest rates of 9% in uncut control plots. Earliest flowering (322 days after planting) and seed formation (411 days) also occurred in plots with no pasture compared with all other pasture treatments (range 1314-1393 days for seed formation to occur). No seeds were produced in uncut plots. At the end of 9 years, mortality rates of bellyache bush plants initially planted averaged 73% for treatments with some pasture compared with 55% under the no-pasture treatment. The percentage of herbaceous plant basal cover in uncut plots was increased 5-fold after 9 years, much greater than the average 2% increase recorded across the low, medium, and high pasture treatments. The number of herbaceous species in uncut plots remained largely unchanged, whereas there was an average reduction of 46% in the cut pasture treatments. Buffel grass remained the species with the greatest basal cover across all cut pasture treatments, followed by sabi grass (Urochloa mosambicensis (Hack.) Dandy) and then red Natal grass (Melinis repens (Willd.) Ziska). These results suggest that grazing strategies that maintain a healthy and competitive pasture layer may contribute to reducing the rate of spread of bellyache bush and complement traditional control techniques such as the use of herbicides.

Natural outbreak of Streptococcus agalactiae (GBS) infection in wild giant Queensland grouper, Epinephelus lanceolatus (Bloch), and other wild fish in northern Queensland, Australia

Ninety-three giant Queensland grouper, Epinephelus lanceolatus (Bloch), were found dead in Queensland, Australia, from 2007 to 2011. Most dead fish occurred in northern Queensland, with a peak of mortalities in Cairns in June 2008. In 2009, sick wild fish including giant sea catfish, Arius thalassinus (Ruppell), and javelin grunter, Pomadasys kaakan (Cuvier), also occurred in Cairns. In 2009 and 2010, two disease epizootics involving wild stingrays occurred at Sea World marine aquarium. Necropsy, histopathology, bacteriology and PCR determined that the cause of deaths of 12 giant Queensland grouper, three wild fish, six estuary rays, Dasyatis fluviorum (Ogilby), one mangrove whipray, Himantura granulata (Macleay), and one eastern shovelnose ray, Aptychotrema rostrata (Shaw), was Streptococcus agalactiae septicaemia. Biochemical testing of 34 S.agalactiae isolates from giant Queensland grouper, wild fish and stingrays showed all had identical biochemical profiles. The 16S rRNA gene sequences of isolates confirmed all isolates were S.agalactiae; genotyping of selected S.agalactiae isolates showed the isolates from giant Queensland grouper were serotype Ib, whereas isolates from wild fish and stingrays closely resembled serotype II. This is the first report of S.agalactiae from wild giant Queensland grouper and other wild tropical fish and stingray species in Queensland, Australia.

Phosphine resistance in Sitophilus oryzae (L.) from eastern Australia: Inheritance, fitness and prevalence

The inheritance and fitness of phosphine resistance was investigated in an Australian strain of the rice weevil, Sitophilus oryzae (L.), as well as its prevalence in eastern Australia. This type of knowledge may provide insights in to the development of phosphine resistance in this species with the potential for better management. This strain was 12.2 × resistant at the LC50 level based on results for adults exposed for 20 h. Data from the testing of F1 adults from the reciprocal crosses (R♀ × S♂ and S♀ × R♂) showed that resistance was autosomal and inherited as an incompletely recessive trait with a degree of dominance of -0.88. The dose-response data for the F1 × S and F1 × R test crosses, and the F2 progeny were compared with predicted dose-response assuming monogenic recessive inheritance, and the results were consistent with resistance being conferred by one major gene. There was no evidence of fitness cost based on the frequency of susceptible phenotypes in hybridized populations that were reared for seven generations without exposure to phosphine. Lack of fitness cost suggests that resistant alleles will tend to persist in field populations that have undergone selection even if selection pressure is removed. Discriminating dose tests on 107 population samples collected from farms from 2006 to 2010 show that populations containing insects with the weak resistant phenotype are common in eastern Australia, although the frequency of resistant phenotypes within samples was typically low. The prevalence of resistance is a warning that this species has been subject to considerable selection pressure and that effective resistance management practices are needed to address this problem. Crown Copyright © 2014.

Field evaluation of transgenic pineapple (Ananas comosus (L.) Merr.) cv. ‘Smooth Cayenne’ for resistance to blackheart under subtropical conditions

A comparative analysis of transgenic pineapple lines transformed with a polyphenol oxidase (PPO) gene (ppo) and the untransformed cultivar ‘Smooth Cayenne’ was made from plants grown in a series of field trials under cool subtropical conditions in southeast Queensland. In the four field trials where blackheart was recorded, all of the control lines expressed blackheart on each occasion and exhibited the greatest incidence (50%) and severity (34%) of symptoms. Irrespective of the gene transfer method or the gene construct used, 38% of the lines produced were regarded as blackheart resistant, having no blackheart symptoms in two or more trials. Five blackheart resistant transgenic lines consistently performed as well as or better than control plants in terms of fruit characteristics and quality.

Resistance gene analogues in sugarcane and sorghum and their association with quantitative trait loci for rust resistance

Fifty-four different sugarcane resistance gene analogue (RGA) sequences were isolated, characterized, and used to identify molecular markers linked to major disease-resistance loci in sugarcane. Ten RGAs were identified from a sugarcane stem expressed sequence tag (EST) library; the remaining 44 were isolated from sugarcane stem, leaf, and root tissue using primers designed to conserved RGA motifs. The map location of 31 of the RGAs was determined in sugarcane and compared with the location of quantitative trait loci (QTL) for brown rust resistance. After 2 years of phenotyping, 3 RGAs were shown to generate markers that were significantly associated with resistance to this disease. To assist in the understanding of the complex genetic structure of sugarcane, 17 of the 31 RGAs were also mapped in sorghum. Comparative mapping between sugarcane and sorghum revealed syntenic localization of several RGA clusters. The 3 brown rust associated RGAs were shown to map to the same linkage group (LG) in sorghum with 2 mapping to one region and the third to a region previously shown to contain a major rust-resistance QTL in sorghum. These results illustrate the value of using RGAs for the identification of markers linked to disease resistance loci and the value of simultaneous mapping in sugarcane and sorghum.

Responses of Super Rice (Oryza sativa L.) to Different Planting Methods for Grain Yield and Nitrogen-Use Efficiency in the Single Cropping Season

To break the yield ceiling of rice production, a super rice project was developed in 1996 to breed rice varieties with super high yield. A two-year experiment was conducted to evaluate yield and nitrogen (N)-use response of super rice to different planting methods in the single cropping season. A total of 17 rice varieties, including 13 super rice and four non-super checks (CK), were grown under three N levels [0 (N0), 150 (N150), and 225 (N225) kg ha−1] and two planting methods [transplanting (TP) and direct-seeding in wet conditions (WDS)]. Grain yield under WDS (7.69 t ha−1) was generally lower than TP (8.58 t ha−1). However, grain yield under different planting methods was affected by N rates as well as variety groups. In both years, there was no difference in grain yield between super and CK varieties at N150, irrespective of planting methods. However, grain yield difference was dramatic in japonica groups at N225, that is, there was an 11.3% and 14.1% average increase in super rice than in CK varieties in WDS and TP, respectively. This suggests that high N input contributes to narrowing the yield gap in super rice varieties, which also indicates that super rice was bred for high fertility conditions. In the japonica group, more N was accumulated in super rice than in CK at N225, but no difference was found between super and CK varieties at N0 and N150. Similar results were also found for N agronomic efficiency. The results suggest that super rice varieties have an advantage for N-use efficiency when high N is applied. The response of super rice was greater under TP than under WDS. The results suggest that the need to further improve agronomic and other management practices to achieve high yield and N-use efficiency for super rice varieties in WDS.

Refining low-volume, high-concentration herbicide applications to control Chromolaena odorata (L.) King & Robinson (Siam weed) in remote areas

Chromolaena odorata (L.) King and Robinson (Siam weed) is a highly invasive plant and a high priority for control in north Queensland. It can be effectively treated using high-volume, groundbased herbicide spray equipment, but operational information shows that this control method becomes increasingly difficult in areas where vehicle access is prevented by rougher terrain. Low-volume, high-concentration herbicide applications have proven capable of causing high mortality in these remote situations. Two trials were undertaken between May 2010 and May 2012 to refine effective rates of aminopyralid/fluroxypyr, fluroxypyr and metsulfuron-methyl, only using low-volume, high-concentration applications on Siam weed. Fluroxypyr on its own was as effective as aminopyralid/fluroxypyr as both herbicides caused 95-100% mortality at overlapping rates containing 5 to 18.85 g a.i. L-1 of fluroxypyr. Metsulfuron-methyl caused 100% mortality when applied at 3 and 6 g a.i. L-1. Effective control was achieved with approximately 16 to 22 mL of the solutions per plant, so a 5 L mixture in a backpack could treat 170 to 310 adult plants. There are several options for treating Siam weed on the ground and the choice of methods reflects the area, plant density and accessibility of the infestation. Control information from Siam weed field crews shows that low volume, high concentration herbicide applications applied using a splatter gun are a more efficient method for controlling larger, denser remote infestations than physical removal. By identifying effective herbicides that are applied through low-volume equipment, these trials provide an additional and more efficient tool for controlling Siam weed in remote areas.

Effects of light conditions and plant density on growth and reproductive biology of Cascabela thevetia (L.) Lippold

Cascabela thevetia (L.) Lippold (Apocynaceae) is an invasive woody weed that has formed large infestations at several locations in northern Australia. Understanding the reproductive biology of C. thevetia is vital to its management. This paper reports results of a shade house experiment that determined the effects of light conditions (100% or 30% of natural light) and plant densities (one, two, four or eight plants per plot) on the growth, time to flowering and seed formation, and monthly pod production of two C. thevetia biotypes (peach and yellow). Shaded plants were significantly larger when they reached reproductive maturity than plants grown under natural light. However, plants grown under natural light flowered earlier (268 days compared with 369 days) and produced 488 more pods per pot (a 5-fold increase) over 3 years. The yellow biotype was slightly taller at reproductive maturity but significantly taller and with significantly greater aboveground biomass at the end of the study. Both biotypes flowered at a similar time under natural light and low plant densities but the yellow biotype was quicker to seed (478 versus 498 days), produced significantly more pods (364 versus 203 pods) and more shoot growth (577 g versus 550 g) than the peach biotype over 3 years. Higher densities of C. thevetia tended to significantly reduce the shoot and root growth by 981 g and 714 g per plant across all light conditions and biotypes over 3 years and increase the time taken to flower by 140 days and produce seeds by 184 days. For land managers trying to prevent establishment of C. thevetia or to control seedling regrowth once initial infestations have been treated, this study indicates that young plants have the potential to flower and produce seeds within 268 and 353 days, respectively. However, with plant growth and reproduction most likely to be slower under field conditions, annual surveillance and control activities should be sufficient to find and treat plants before they produce seeds and replenish soil seed banks. The most at-risk part of the landscape may be open areas that receive maximum sunlight, particularly within riparian habitats where plants would consistently have more favourable soil moisture conditions.