Effect of abscisic acid pretreatments on stomatal reopening in Vicia faba

After a pretreatment of 2 h exposure to a solution containing 2 × 10⁻⁴ M ABA, reopening of stomata occurred in epidermal strips of Vicia faba L. cv. Cavalier on an ABA-free incubation solution. After pretreatment with exogenous ABA stomatal apertures were greater when higher levels of KCl were incorporated into the solution used for reopening. Prolonged exposure to exogenous ABA (14 h) did not prevent stomatal reopening upon transfer to ABA-free solutions. However, for both ABA and ABA-free pretreatments, prolonged incubation (1 day after removal of epidermis) resulted in enhanced stomatal apertures when the epidermal strips were exposed to light. This effect was lost 2 days after removal of the epidermis and opening did not occur after 3 days. Epidermal strips containing endogenous ABA were obtained from wilted leaves. Reopening was greatly reduced by the endogenous ABA treatment, and variation of KCl concentration in the incubation solution had little effect on stomatal aperture. It is postulated that during wilting endogenous ABA becomes reversibly bound without loss of activity for a longer period than is obtained using exogenous ABA. The presence of other unidentified compounds may be involved in this process.     

Adelphocorisella Miyamoto and Yasunaga newly recorded from Australia, with the description of a new species (Heteroptera: Miridae: Mirinae)

Adelphocorisella australis sp. n. is described from north Queensland. This, the first representative of the genus known from Australia, is compared with the two previously described species, both from Japan.     

Rapid "open-source" engineering of customized zinc-finger nucleases for highly efficient gene modification

Custom-made zinc-finger nucleases (ZFNs) can induce targeted genome modifications with high efficiency in cell types including Drosophila, C. elegans, plants, and humans. A bottleneck in the application of ZFN technology has been the generation of highly specific engineered zinc-finger arrays. Here we describe OPEN (Oligomerized Pool ENgineering), a rapid, publicly available strategy for constructing multifinger arrays, which we show is more effective than the previously published modular assembly method. We used OPEN to construct 37 highly active ZFN pairs which induced targeted alterations with high efficiencies (1%-50%) at 11 different target sites located within three endogenous human genes (VEGF-A, HoxB13, and CFTR), an endogenous plant gene (tobacco SuRA), and a chromosomally integrated EGFP reporter gene. In summary, OPEN provides an "open-source" method for rapidly engineering highly active zinc-finger arrays, thereby enabling broader practice, development, and application of ZFN technology for biological research and gene therapy.     

Targeted mutagenesis of duplicated genes in soybean with zinc-finger nucleases

We performed targeted mutagenesis of a transgene and nine endogenous soybean (Glycine max) genes using zinc-finger nucleases (ZFNs). A suite of ZFNs were engineered by the recently described context-dependent assembly platform--a rapid, open-source method for generating zinc-finger arrays. Specific ZFNs targeting dicer-like (DCL) genes and other genes involved in RNA silencing were cloned into a vector under an estrogen-inducible promoter. A hairy-root transformation system was employed to investigate the efficiency of ZFN mutagenesis at each target locus. Transgenic roots exhibited somatic mutations localized at the ZFN target sites for seven out of nine targeted genes. We next introduced a ZFN into soybean via whole-plant transformation and generated independent mutations in the paralogous genes DCL4a and DCL4b. The dcl4b mutation showed efficient heritable transmission of the ZFN-induced mutation in the subsequent generation. These findings indicate that ZFN-based mutagenesis provides an efficient method for making mutations in duplicate genes that are otherwise difficult to study due to redundancy. We also developed a publicly accessible Web-based tool to identify sites suitable for engineering context-dependent assembly ZFNs in the soybean genome.     

Collaborative research partnerships inform monitoring and management of aquatic ecosystems by Indigenous rangers

Aquatic ecosystems are critical to the long-term viability and vibrancy of communities and economies across northern Australia. In a region that supports significant cultural and ecological water values, partnerships between Indigenous and non-Indigenous stakeholders can benefit aquatic ecosystem management. We present, as a case study from the Kimberley region of Western Australia, a collaborative research program that successfully documented Indigenous and Western Scientific knowledge of remote wetlands, using a variety of field-based activities, questionnaires, interviews and workshops. The sharing of knowledge between Indigenous and non-Indigenous research partners facilitated a comprehensive understanding of ecosystem values, threats, processes, management priorities and aspirations. These formed the basis of a management plan and monitoring tools, designed to build the capacity of an Indigenous ranger group to engage in research, monitoring and management of wetlands. The project provides a useful example of the benefits of collaborations in the context of remote-area management where local communities are responsible for environmental management and monitoring, such as is the case in northern Australia and presumably other areas of the world.