In Vitro Conditions for a Successful Biolistics transformation System of Pineapples (ananas comosus merr.), in 'Contributing to a Sustainable Future'

In order to develop an efficient and reliable biolistics transformation system for pineapples parameters need to be optimised for growth, survival and development of explants pre- and post transformation. We have optimised in vitro conditions for culture media for the various stages of plant and callus initiation and development, and for effective selection of putative transgenic material. Shoot multiplication and proliferation is best on medium containing MS basic nutrients and vitamins with the addition of 0.1 mg/L myo-inositol, 20 g/L sucrose, 2.5 mg/L BAP and 3 g/L Phytagel, followed by transfer to basic MS medium for further development. Callus production on leaf base explants is best on MS nutrients and vitamins, to which 10 mg/L of BAP and NAA each was added. Optimum explant age for bombardment is 17-35 week old callus, while a pre-bombardment osmoticum treatment in the medium is not required. By comparing several antibiotics as selective agent, it has been established that a two-step selection of 2 fortnightly sub-cultures on 50 μg/mL of geneticin in the culture medium, followed by monthly sub-cultures on 100 μg/mL geneticin is optimal for survival of transgenic callus. Shoot regeneration from callus cultures is optimal on medium containing MS nutrients and vitamins, 5% coconut water and 400 mg/L casein hydrolysate. Plants can be readily regenerated and multiplied from transgenic callus through organogenesis. Rooting of shoots does not require any additional plant hormones to the medium. A transformation efficiency of 1 – 3.5% can be achieved, depending on the gene construct applied.

Factors influencing the release and establishment of weed biological control agents

Mike Day and colleagues recently published their paper 'Factors influencing the release and establishment of weed biocontrol agents' in Proceedings of the 16th Australian Weeds Conference. The CRC for Australian Weed Management facilitated an investigation into the factors influencing the release and establishment of weed biological control agents on a wide variety of Australian weeds. The investigation improved the understanding of post-release ecology of biocontrol agents and generated recommendations for best practice. Factors affecting successful establishment on the weed include host plant characteristics, size of releases, dispersal power of the agent, predation and parasitism, and climate. A best practice guide was produced by the CRC to assist practitioners in designing robust release strategies to increase rates of establishment.

Defining critical soil nutrient concentrations in soils supporting grains and cotton in Northern NSW and Queensland

This project focussed on the phosphorus (P) and potassium (K) status of northern cropping soils. Stores of P and K have been depleted by crop removal and limited fertiliser application, with depletion most significant in the subsoil. Soil testing strategies are confounded by slowly available mineral reserves with uncertain availability. The utility of new soil tests was assessed to measure these reserves, their availability to plants quantified and a regional sampling strategy undertaken to identify areas of greatest P and K deficit. Fertiliser application strategies for P and K have been tested and the interactions between these and other nutrients have been determined in a large field program.

Influence of different nitrogen rates and DMPP nitrification inhibitor on annual N2/O emissions from a subtropical wheat–maize cropping system

Global cereal production will need to increase by 50% to 70% to feed a world population of about 9 billion by 2050. This intensification is forecast to occur mostly in subtropical regions, where warm and humid conditions can promote high N2O losses from cropped soils. To secure high crop production without exacerbating N2O emissions, new nitrogen (N) fertiliser management strategies are necessary. This one-year study evaluated the efficacy of a nitrification inhibitor (3,4-dimethylpyrazole phosphate—DMPP) and different N fertiliser rates to reduce N2O emissions in a wheat–maize rotation in subtropical Australia. Annual N2O emissions were monitored using a fully automated greenhouse gas measuring system. Four treatments were fertilized with different rates of urea, including a control (40 kg-N ha−1 year−1), a conventional N fertiliser rate adjusted on estimated residual soil N (120 kg-N ha−1 year−1), a conventional N fertiliser rate (240 kg-N ha−1 year−1) and a conventional N fertiliser rate (240 kg-N ha−1 year−1) with nitrification inhibitor (DMPP) applied at top dressing. The maize season was by far the main contributor to annual N2O emissions due to the high soil moisture and temperature conditions, as well as the elevated N rates applied. Annual N2O emissions in the four treatments amounted to 0.49, 0.84, 2.02 and 0.74 kg N2O–N ha−1 year−1, respectively, and corresponded to emission factors of 0.29%, 0.39%, 0.69% and 0.16% of total N applied. Halving the annual conventional N fertiliser rate in the adjusted N treatment led to N2O emissions comparable to the DMPP treatment but extensively penalised maize yield. The application of DMPP produced a significant reduction in N2O emissions only in the maize season. The use of DMPP with urea at the conventional N rate reduced annual N2O emissions by more than 60% but did not affect crop yields. The results of this study indicate that: (i) future strategies aimed at securing subtropical cereal production without increasing N2O emissions should focus on the fertilisation of the summer crop; (ii) adjusting conventional N fertiliser rates on estimated residual soil N is an effective practice to reduce N2O emissions but can lead to substantial yield losses if the residual soil N is not assessed correctly; (iii) the application of DMPP is a feasible strategy to reduce annual N2O emissions from sub-tropical wheat–maize rotations. However, at the N rates tested in this study DMPP urea did not increase crop yields, making it impossible to recoup extra costs associated with this fertiliser. The findings of this study will support farmers and policy makers to define effective fertilisation strategies to reduce N2O emissions from subtropical cereal cropping systems while maintaining high crop productivity. More research is needed to assess the use of DMPP urea in terms of reducing conventional N fertiliser rates and subsequently enable a decrease of fertilisation costs and a further abatement of fertiliser-induced N2O emissions.

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.

First report of Pythiogeton ramosum (Pythiales) in Australia

In 2012, a project was initiated to assess if the soft rot disease of ginger in Australian fields was associated with pathogens other than Pythium myriotylum. Together with nine Pythium spp., ten isolates of a Pythium-like organism were also recovered from ginger with soft rot symptoms. These Pythium-like isolates were identified as Pythiogeton (Py.) ramosum based on its morphology and ITS sequences. In-vitro pathogenicity tests allowed confirmation of pathogenicity of Py. ramosum on excised carrot (Daucus carota), sweet potato (Ipomoea batatas) and potato (Solanum tubersum) tubers, although it was not pathogenic on excised ginger (Zingiber officinale) and radish (Raphanus sativus) rhizome/roots. In addition it was found to be pathogenic on bean (Phaseolus vulgaris), capsicum (Capsicum annuum) and cauliflower (Brassica oleracea var. botrytis) seedlings. This is the first record of Py. ramosum and its pathogenicity in Australia.

Controlling Pythium in Ginger: Phase 2

The fungal disease Pythium Soft Rot is regarded by the ginger industry as their most serious disease threat. This project was developed to further investigate the factors contributing to the persistence and spread of Pythium Soft Rot on ginger farms and to identify measures for their control. The study demonstrated that the pathogen capable of causing Pythium Soft Rot in ginger was spread in contaminated ‘seed’, soil and water and that it can be managed through a combination of strategies that rely on early detection, reducing pathogen levels in soils, preventing water logging and restricting movement of contaminated ‘seed’, soil and water. However, in order to have an effective level of control, all strategies need to be integrated in an effective manner.

Pythium soft rot of ginger: Detection and identification of the causal pathogens, and their control

Ginger is considered by many people to be the outstanding member among 1400 other species in the family Zingiberaceae. Not only it is a valuable spice used by cooks throughout the world to impart unique flavour to their dishes but it also has a long track record in some Chinese and Indian cultures for treating common human ailments such as colds and headaches. Ginger has recently attracted considerable attention for its anti-inflammatory, antibacterial and antifungal properties. However, ginger as a crop is also susceptible to at least 24 different plant pathogens, including viruses, bacteria, fungi and nematodes. Of these, Pythium spp. (within the kingdom Stramenopila, phyllum Oomycota) are of most concern because various species can cause rotting and yield loss on ginger at any of the growth stages including during postharvest storage. Pythium gracile was the first species in the genus to be reported as a ginger pathogen, causing Pythium soft rot disease in India in 1907. Thereafter, numerous other Pythium spp. have been recorded from ginger growing regions throughout the world. Today, 15 Pythium species have been implicated as pathogens of the soft rot disease. Because accurate identification of a pathogen is the cornerstone of effective disease management programs, this review will focus on how to detect, identify and control Pythium spp. in general, with special emphasis on Pythium spp. associated with soft rot on ginger.

Pythiogeton ramosum, a new pathogen of soft rot disease of ginger (Zingiber officinale) at high temperatures in Australia

Pythium soft rot (PSR) of ginger caused by a number of Pythium species is of the most concern worldwide. In Australia, PSR outbreaks associated with Pythium myriotylum was recorded in 2007. Our recent pathogenicity tests in Petri dishes conducted on ginger rhizomes and pot trials on ginger plants showed that Pythiogeton (Py.) ramosum, an uncommon studied oomycete in Pythiaceae, was also pathogenic to ginger at high temperature (30–35 °C). Ginger sticks excised from the rhizomes were colonised by Py. ramosum which caused soft rot and browning lesions. Ginger plants inoculated with Py. ramosum showed initial symptoms of wilting and leave yellowing, which were indistinguishable from those of Pythium soft rot of ginger, at 10 days after inoculation. In addition, morphological and phylogenetic studies indicated that isolates of Py. ramosum were quite variable and our isolates obtained from soft rot ginger were divided into two groups based on these variations. This is also for the first time Py. ramosum is reported as a pathogen on ginger at high temperatures.

Legumes or nitrification inhibitors to reduce N2O emissions from subtropical cereal cropping systems in Oxisols?

The DAYCENT biogeochemical model was used to investigate how the use of fertilizers coated with nitrification inhibitors and the introduction of legumes in the crop rotation can affect subtropical cereal production and N2O emissions. The model was validated using comprehensive multi-seasonal, high-frequency dataset from two field investigations conducted on an Oxisol, which is the most common soil type in subtropical regions. Different N fertilizer rates were tested for each N management strategy and simulated under varying weather conditions. DAYCENT was able to reliably predict soil N dynamics, seasonal N2O emissions and crop production, although some discrepancies were observed in the treatments with low or no added N inputs and in the simulation of daily N2O fluxes. Simulations highlighted that the high clay content and the relatively low C levels of the Oxisol analyzed in this study limit the chances for significant amounts of N to be lost via deep leaching or denitrification. The application of urea coated with a nitrification inhibitor was the most effective strategy to minimize N2O emissions. This strategy however did not increase yields since the nitrification inhibitor did not substantially decrease overall N losses compared to conventional urea. Simulations indicated that replacing part of crop N requirements with N mineralized by legume residues is the most effective strategy to reduce N2O emissions and support cereal productivity. The results of this study show that legumes have significant potential to enhance the sustainable and profitable intensification of subtropical cereal cropping systems in Oxisols.

Sorption and desorption of sulfuryl fluoride by wheat, flour and semolina

Sulfuryl fluoride (SF) has been developed as a fumigant for control of insect pests in stored grain. However, there is very limited information on the sorption behaviour of this fumigant, which can be critical to its bioactivity, application and potential for residues. We undertook a comprehensive laboratory study of the sorption and desorption of SF by wheat (bread and durum), flour and semolina at 15, 25 and 35 °C, moisture contents 12% and 15%, and concentration × time combinations at CT = 1500 mgh/L (4.167 mg/L × 360 h, 8.928 mg/L × 168 h and 31.25 mg/L × 48 h). At each dosage, sorption rate increased as commodity temperature and moisture content increased. The highest rates of sorption occurred at 35 °C and 15% m.c., and lowest rates at 15 °C and 12% m.c., and the rate was independent of initial concentration. Sorption followed first order reaction kinetics described by the exponential decay equation, Ct = C0·e−k*t, where k is the sorption rate constant. The most important factors determining the rate of sorption were commodity particle size (exposed surfaces) and temperature. Little sorption of fumigant occurred within the first 24 h whereas longer fumigation times resulted in significant sorption. Unbound SF was rapidly lost from the commodity upon aeration with no further desorption detected under any of the test conditions.

Does Fumigation of Durum Wheat and Semolina with Sulfuryl Fluoride affect Quality of the Grain, Semolina and derived Spaghetti and Bread?

There is no information on the effect of sulfuryl fluoride (SF) on durum wheat technological properties and products made from fumigated durum wheat. Durum wheat and semolina were exposed to a range of SF applications under conditions that might be typically encountered in bulk storage facilities used in many countries. SF greatly reduced the germination percentage of fumigated durum wheat with increasing impact under higher SF concentration, grain moisture content, and fumigation temperature. SF greatly reduced seed germination percentage impacting more the higher the SF concentration. SF had little to no effects on grain test weight, 1000 grain weight, hardness, protein content, semolina ash content and mixograph properties. At the highest SF concentration (31.25 mg/L for 48 h) there was a tendency for pasta cooking loss to be increased but still acceptable while other pasta properties were largely unaffected. Fumigation with SF did not have any impact on the baking properties of a wholemeal durum flour-commercial flour mix. Therefore, SF is not recommended if the grains are to be used as seeds for agricultural production but for the production of semolina, pasta and bread, SF used under typical fumigation conditions has little to no impact on technological properties of durum wheat.