Ginger (Zingiber officinale) autotetraploids with improved processing quality produced by an in vitro colchicine treatment

Ginger autotetraploids were produced by immersing shoot tips in a 0.5% w/v colchicine, 2% v/v dimethyl sulfoxide solution for 2 h. Stomatal measurements were used as an early indicator of ploidy differences in culture with mean stomata length of tetraploids (49.2 μm) being significantly larger than the diploid (38.8 µm). Of the 500 shoot tips treated, 2% were characterised as stable autotetraploid lines following field evaluation over several seasons. Results were confirmed with flow cytometry and, of the 7 lines evaluated for distinctness and uniformity, 6 were solid tetraploid mutants and 1 was a periclinal chimera. Significant differences were noted between individual tetraploid lines in terms of shoot length, leaf length, leaf width, size of rhizome sections (knob weight) and fibre content. The solid autotetraploid lines had significantly wider, greener leaves than the diploids, they had significantly fewer but thicker shoots and, although ‘Queensland’ (the diploid parent from which the tetraploids were derived) had a greater total rhizome mass at harvest, its knob size was significantly smaller. From the autotetraploid lines, one line was selected for commercial release as ‘Buderim Gold’. It compared the most favourably with ‘Queensland’ in terms of the aroma/flavour profile and fibre content at early harvest, and had consistently good rhizome yield. More importantly it produced large rhizome sections, resulting in a higher recovery of premium grade confectionery ginger and a more attractive fresh market product.

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.

Insecticide resistance and implications for future aphid management in Australian grains and pastures: A review

Aphids can cause substantial damage to cereals, oilseeds and legumes through direct feeding and through the transmission of plant pathogenic viruses. Aphid-resistant varieties are only available for a limited number of crops. In Australia, growers often use prophylactic sprays to control aphids, but this strategy can lead to non-target effects and the development of insecticide resistance. Insecticide resistance is a problem in one aphid pest of Australian grains in Australia, the green peach aphid (Myzus persicae). Molecular analyses of field-collected samples demonstrate that amplified E4 esterase resistance to organophosphate insecticides is widespread in Australian grains across Australia. Knockdown resistance to pyrethroids is less abundant, but has an increased frequency in areas with known frequent use of these insecticides. Modified acetylcholinesterase resistance to dimethyl carbamates, such as pirimicarb, has not been found in Australia, nor has resistance to imidacloprid. Australian grain growers should consider control options that are less likely to promote insecticide resistance, and have reduced impacts on natural enemies. Research is ongoing in Australia and overseas to provide new strategies for aphid management in the future.

A core metabolic enzyme mediates resistance to phosphine gas

Phosphine is a small redox-active gas that is used to protect global grain reserves, which are threatened by the emergence of phosphine resistance in pest insects. We find that polymorphisms responsible for genetic resistance cluster around the redox-active catalytic disulfide or the dimerization interface of dihydrolipoamide dehydrogenase (DLD) in insects (Rhyzopertha dominica and Tribolium castaneum) and nematodes (Caenorhabditis elegans). DLD is a core metabolic enzyme representing a new class of resistance factor for a redox-active metabolic toxin. It participates in four key steps of core metabolism, and metabolite profiles indicate that phosphine exposure in mutant and wild-type animals affects these steps differently. Mutation of DLD in C. elegans increases arsenite sensitivity. This specific vulnerability may be exploited to control phosphine-resistant insects and safeguard food security.

The insecticidal spider toxin SFI1 is a knottin peptide that blocks the pore of insect voltage-gated sodium channels via a large β-hairpin loop

Spider venoms contain a plethora of insecticidal peptides that act on neuronal ion channels and receptors. Because of their high specificity, potency and stability, these peptides have attracted much attention as potential environmentally friendly insecticides. Although many insecticidal spider venom peptides have been isolated, the molecular target, mode of action and structure of only a small minority have been explored. Sf1a, a 46-residue peptide isolated from the venom of the tube-web spider Segesteria florentina, is insecticidal to a wide range of insects, but nontoxic to vertebrates. In order to investigate its structure and mode of action, we developed an efficient bacterial expression system for the production of Sf1a. We determined a high-resolution solution structure of Sf1a using multidimensional 3D/4D NMR spectroscopy. This revealed that Sf1a is a knottin peptide with an unusually large β-hairpin loop that accounts for a third of the peptide length. This loop is delimited by a fourth disulfide bond that is not commonly found in knottin peptides. We showed, through mutagenesis, that this large loop is functionally critical for insecticidal activity. Sf1a was further shown to be a selective inhibitor of insect voltage-gated sodium channels, consistent with its 'depressant' paralytic phenotype in insects. However, in contrast to the majority of spider-derived sodium channel toxins that function as gating modifiers via interaction with one or more of the voltage-sensor domains, Sf1a appears to act as a pore blocker.