Climatic Drivers of Revegetation Management Practices in Australia: Analysis of a Social Survey

Investment in small and large-scale revegetation in Australia is growing in response to concerns regarding the sustainability and productivity of agricultural landscapes. Site preparation and management––such as soil cultivation, weed control, fertilising, mulching, use of treeguards and watering––are major costs associated with small-scale revegetation. The aim of this study has been to investigate local revegetation knowledge and practices to determine the usefulness of each management practice for achieving success and to determine whether some practices are more suited to particular climatic zones. A national online revegetation survey was conducted to ascertain current small-scale revegetation practices and the factors that drive these choices. Management practices were found to be strongly associated with climate. Mulch, fertiliser, weed control and watering were applied more frequently in higher rainfall and higher temperature zones. Soil cultivation and treeguards were used more frequently in lower rainfall and lower temperature zones. These findings suggest that there may be some benefit in modifying existing revegetation guidelines to reflect climatic zones and management flexibility.

Proteomic analysis of membrane preparations from developing Pinus radiata compression wood

For coniferous gymnosperms, few data exist as to the contribution of the membrane-associated proteome to cell wall and wood formation. In this study, we begin to address this knowledge deficiency by examining the proteomic profile of Golgi-enriched membrane preparations derived from developing Pinus radiata compression wood. These membrane preparations were generated by a combination of discontinuous sucrose gradient centrifugation and Triton X-114-based phase separation. Fractionation by phase separation removed contaminating proteins associated with the cytoskeleton and enabled the discrimination between soluble and membrane-bound/integral proteins. The proteomic analysis of the resulting aqueous and detergent phases using high-performance liquid chromatography-tandem mass spectrometry resulted in the identification of 175 proteins. The majority of the identified proteins were membrane bound/integral and originated from cellular components such as the nucleus, plastids, endoplasmic reticulum, plasma membrane and Golgi vesicles. On the basis of bioinformatic analysis, many of the identified proteins were predicted to be involved either in the regulation of wood formation or in cell wall biosynthesis, which indicated that the proteomic analysis of non-cytosolic proteins in developing xylem is a useful strategy to investigate the molecular aspects of wood formation in pine.     

Detecting the undetectable: Characterization,optimization, and validation of an eDNA detection assay for the federally endangered dwarf wedgemussel,Alasmidonta heterodon (Bivalvia: Unionoida)

1. Environmental DNA (eDNA) assays are valuable tools for monitoring the presence and distribution of cryptic species.
2. Like many freshwater mussels, the numbers of dwarf wedgemussel, Alasmidonta heterodon, have dwindled and its range has diminished. As of its listing in 1993, only 10–20 locations were known to persist out of the 70 Atlantic slope locations known historically.
3. A quantitative polymerase chain reaction (qPCR) assay to detect the presence of A. heterodon was developed that uses two probes to accommodate a single‐nucleotide polymorphism (SNP) in the probe‐binding site within the cytochrome c oxidase subunit I (COI) gene. This SNP defines northern and southern major phylogenetic lineages.
4. The primers match exactly the previously determined COI sequences of 20 dwarf wedgemussel individuals representing Atlantic slope populations from North Carolina, Virginia, Maryland, New York, and New Hampshire. Other than for the qPCR assay described here, these primers can be used for sequencing or metabarcoding to further delineate dwarf wedgemussel populations phylogenetically.
5. A simple eDNA preparation method is introduced using flocculation to concentrate free DNA in solution as well as cellular material (including shed animal cells, bacteria, viruses, and dissolved DNA).
6. In addition to the specific application described here, the methodological approaches used in this study are widely applicable to conservation, including, but not limited to, general aquatic biodiversity, phylogenetic studies, and the detection of pathogenic microbes.

     

Gonadal Stage and Sex Steroid Correlations in Siberian Sturgeon,Acipenser baeri,Habituated to a Semitropical Environment

Stages of gonadal development, in association with plasma concentrations of the sex steroids 17β‐estradiol (E2), progesterone (P4), testosterone (T), and 11‐ketotestosterone (11‐KT), were investigated for a single time point during a natural breeding season in 7‐yr‐old Siberian sturgeon, Acipenser baeri Brandt, exposed lifelong to a warmwater environment. Among females, examination of gonadal tissue showed variation in ovarian stage, with 12.5, 47.5, 22.5, and 17.5% of females found at Stages 2 (previtellogenic), 3 (early vitellogenic), 4 (mid‐vitellogenic), and 5 (migratory nucleus), respectively. Although patterns varied among the hormones, plasma concentrations of E2, T, and 11‐KT became increasingly elevated in females as maturation progressed. On the basis of histological criteria, males were classified as either premeiotic (quiescent) or meiotic and 50% of the males sampled were found at each stage. Significant elevations in circulating concentrations of plasma E2 and T were observed in meiotic versus premeiotic males, and there was a rise in plasma 11‐KT concentration that approached significance (P = 0.056).     

Molecular characterization of a stable antisense chalcone synthase phenotype in strawberry (Fragaria x ananassa)

An octaploid (Fragaria x ananassa cv. Calypso) genotype of strawberry was transformed with an antisense chalcone synthase (CHS) gene construct using a ripening related CHS cDNA from Fragaria x ananassa cv. Elsanta under the control of the constitutive CaMV 35S promoter via Agrobacterium tumefaciens. Out of 25 transgenic lines, nine lines showed a reduction in CHS mRNA accumulation of more than 50% as compared to the untransformed cv. Calypso control. The antisense CHS construct was found to be integrated into the genome, with a copy number ranging from one to four. The pigmentation of the fruit was only affected when less than 5% of the control CHS expression level was detected. A stable antisense phenotype over a period of 4 years was obtained in the primary transgenic lines at a rate of 1:20. As a consequence of the reduced activity of CHS, the levels of anthocyanins, flavonols, and proanthocyanidins were downregulated and precursors of the flavonoid pathway were shunted to the phenylpropanoid pathway leading to highly increased levels of cinnamoyl glucose (520% of control), caffeoyl glucose (816% of control), and feruloyl glucose (1092% of control) as well as p-coumaryl alcohol (363% of control) and p-coumaryl-1-acetate (1079% of control), which occur only as trace components in untransformed control fruits. These results demonstrate that the introduction of an antisense CHS construct in strawberry results in an unpredictable biochemical phenotype, thereby confirming that CHS function is an important regulatory point of substrate flow between the flavonoid and the phenylpropanoid pathways.     

Denitrification potential of intermittently saturated floodplain soils from a subtropical perennial stream and an ephemeral tributary

Denitrification has the potential to remove excess nitrogen from groundwater passing through riparian buffers, thus improving water quality downstream. In regions with markedly seasonal precipitation, transient stream flow events may be important in saturating adjacent floodplain soils and intermittently providing the anaerobic conditions necessary for denitrification to occur. In two experiments we characterised the denitrification potential of soils from two contrasting floodplains that experience intermittent saturation. We quantified under controlled laboratory conditions: 1) potential rates of denitrification in these soils with depth and over time, for a typical period of saturation; and 2) the influences on rates of nitrate and organic carbon. Treatments differed between experiments, but in each case soil-water slurries were incubated anaerobically with differing amendments of organic carbon and nitrate; denitrification rates were measured at selected time intervals by the acetylene-block technique; and slurry filtrates were analysed for various chemical constituents. In the first experiment (ephemeral tributary), denitrification was evident in soils from both depths (0-0.3 m; 0.3-1.1 m) within hours of saturation. Before Day 2, mean denitrification rates at each depth were generally comparable, irrespective of added substrates; mean rates (Days 0 and 1) were 5.2 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0-0.3 m) and 1.6 ± 0.2 mg N kg dry soil⁻¹ day⁻¹ (0.3-1.1 m). Rates generally peaked on Days 2 or 3. The availability of labile organic carbon was a major constraint on denitrification in these soils. Acetate addition greatly increased rates, reaching a maximum in ephemeral floodplain soils of 17.4 ± 1.8 mg N kg dry soil⁻¹ day⁻¹ on Day 2: in one deep-soil treatment (low nitrate) this overcame differences in rates observed with depth when acetate was not added, although the rate increase in the other deep-soil treatment (high nitrate) was significantly less (P ≤ 0.01). Without acetate, peak denitrification rates in this experiment were 6.9 ± 0.4 and 2.8 ± 0.2 mg N kg dry soil⁻¹ day⁻¹ in surface and deep soils, respectively. Differences in rates were observed with depth on all occasions, despite similar initial concentrations of dissolved organic carbon (DOC) at both depths. Levels of substrate addition in the second experiment (perennial stream) more closely reflected natural conditions at the site. Mean denitrification rates were consistently much higher in surface soil (P ≤ 0.001), while the source of water used in the slurries (surface water or groundwater from the site) had little effect on rates at any depth. Mean rates when all treatments retained nitrate were: 4.5 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0-0.3 m depth); 0.8 ± 0.3 mg N kg dry soil⁻¹ day⁻¹ (0.3-1.0 m); and 0.6 ± 0.1 mg N kg dry soil⁻¹ day⁻¹ (1.8-3.5 m). For comparable treatments and soil depths, denitrification potentials at both sites were similar, apart from higher initial rates in the ephemeral floodplain soils, probably associated with their higher DOC content and possibly also their history of more frequent saturation. The rapid onset of denitrification and the rates measured in these soils suggest there may be considerable potential for nitrate removal from groundwater in these floodplain environments during relatively short periods of saturation.