Accelerator trial series in Pinus radiata stands in New Zealand: Trial establishment, site description and initial soil, forest floor and tree data.

Interest in establishing biological-based economies has created increasing and rapidly moving demand for wood and fibre from production forests. Meeting the global demand for timber supply will require investment and development across all components of the supply chain but will ultimately rely on the ability of the forestry sector to increase productivity without compromising the sustainability of plantation management. To address this issue in the context of New Zealand forestry, a trial series was established from 2015 to 2018 to accelerate plantation forest growth by exploring current and future limitations to timber productivity, then altering management practices to overcome these limits. The six sites in this Accelerator trial series were planted with a mix of 12 different types of Pinus radiata D. Don stock expressing various traits related to tree growth, health and wood quality. The planting stock included ten clones, a hybrid and a seed lot representing a widely planted tree stock used throughout New Zealand. At each trial site a range of treatments were applied, including a control. The treatments were designed to address the specific current and predicted limitations to productivity at each location, with consideration for environmental sustainability and impacts on wood quality. Additional site-specific treatments will be implemented across the approximately 30-year life span of each trial. Here we present data describing both the pre-harvest and time zero state of at each trial site. These data provide a baseline that will enable treatment responses to be holistically understood as the trial series matures. This comparison will determine if current tree productivity has been enhanced, and if improvements in site characteristics may also benefit future rotations. The Accelerator trials represent an ambitious research goal that will take planted forest productivity to a new level of enhanced long-term forest productivity without compromising the sustainable management of future forests.

Metal biosorption in lignocellulosic biofuel biorefinery effluent: an initial step towards sustainability of water resources.

Biosorption of metals by microorganisms is a promising technology to remove accumulated non-process elements in highly recycled biorefinery process water. Removal of these elements would enable greater water reuse and reduce the environmental impact of effluent discharge. A model lignocellulosic ethanol biorefinery wastewater was created based on pulp mill effluent. This generated a wastewater with an environmentally realistic high loading of dissolved natural organic matter (900 mg/l), a potentially important factor influencing metal biosorption. Analysis of feedstock and pulp mill effluent indicated that Mn and Zn are likely to be problematic in highly recycled lignocellulosic ethanol biorefinery process water. Therefore, the growth of several bacteria and fungi from existing collections, and some isolated from pulp mill effluent were tested in the model wastewater spiked with Mn and Zn (0.2 mM). Wastewater isolates grew the best in the wastewater. Metal uptake varied by species and was much greater for Zn than Mn. A bacterium, Novosphingobium nitrogenifigens Y88(T), removed the most metal per unit biomass, 35 and 17 mg Mn/g. No other organism tested decreased the Mn concentration. A yeast, Candida tropicalis, produced the most biomass and removed the most total metal (38 % of Zn), while uptake per unit biomass was 24 mg Zn/g. These results indicate that microorganisms can remove significant amounts of metals in wastewater with high concentrations of dissolved natural organic matter. Metal sorption by autochthonous microorganisms in an anaerobic bioreactor may be able to extend water reuse and therefore lower the water consumption of future biorefineries.

Biodiversity of active and inactive bacteria in the gut flora of wood-feeding huhu beetle larvae (Prionoplus reticularis).

Huhu grubs (Prionoplus reticularis) are wood-feeding beetle larvae endemic to New Zealand and belonging to the family Cerambycidae. Compared to the wood-feeding lower termites, very little is known about the diversity and activity of microorganisms associated with xylophagous cerambycid larvae. To address this, we used pyrosequencing to evaluate the diversity of metabolically active and inactive bacteria in the huhu larval gut. Our estimate, that the gut harbors at least 1,800 phylotypes, is based on 33,420 sequences amplified from genomic DNA and reverse-transcribed RNA. Analysis of genomic DNA- and RNA-derived data sets revealed that 71% of all phylotypes (representing 95% of all sequences) were metabolically active. Rare phylotypes contributed considerably to the richness of the community and were also largely metabolically active, indicating their participation in digestive processes in the gut. The dominant families in the active community (RNA data set) included Acidobacteriaceae (24.3%), Xanthomonadaceae (16.7%), Acetobacteraceae (15.8%), Burkholderiaceae (8.7%), and Enterobacteriaceae (4.1%). The most abundant phylotype comprised 14% of the active community and affiliated with Dyella ginsengisoli (Gammaproteobacteria), suggesting that a Dyella-related organism is a likely symbiont. This study provides new information on the diversity and activity of gut-associated microorganisms that are essential for the digestion of the nutritionally poor diet consumed by wood-feeding larvae. Many huhu gut phylotypes affiliated with insect symbionts or with bacteria present in acidic environments or associated with fungi.

Identifying diazotrophs by incorporation of nitrogen from (15)N(2) into RNA.

The diversity and abundance of active diazotrophs was investigated in a New Zealand pulp and paper wastewater by enrichment with (15)N(2). Purified (15)N-RNA was analysed by reverse transcription, molecular cloning and sequence analysis of 16S rRNA to reveal a diverse community of bacteria as indicated by a Shannon Weaver Index value of > 2.8. The major class represented in the enriched culture were the gamma-Proteobacteria at 85% with a secondary group of the phylum Firmicutes present at 8.2%, the remaining sequences were affiliated with the alpha- and beta-Proteobacterial classes (1.4% and 4.3%, respectively). Three dominant genera, Aeromonas, Pseudomonas and Bacillus, were identified by comparison with published sequences and phylogenetic analysis. To confirm that representatives of the taxonomic groups identified from the active enriched nitrogen-fixing community were capable of fixing nitrogen Aeromonas and Pseudomonas species were cultivated and shown to possess nifH genes. In wastewater, fluorescence in situ hybridisation probing revealed that the dominant nitrogen-fixing population identified in this study were present in the population, but at lower levels. The population is, therefore, reliant on a small sub-population of diazotrophs to supply the community's nitrogen needs above that already present in the wastewater.

The bacterial microbiota of Stolotermes ruficeps (Stolotermitidae), a phylogenetically basal termite endemic to New Zealand.

Stolotermes ruficeps is a widespread, primitive, lower termite occupying dead and decaying wood of many tree species in New Zealand's temperate forests. We identified core bacterial taxa involved in gut processes through combined DNA- and RNA (cDNA)-based pyrosequencing analysis of the 16S nucleotide sequence from five S. ruficeps colonies. Most family and many genus-level taxa were common to S. ruficeps colonies despite being sampled from different tree species. Major taxa identified were Spirochaetaceae, Elusimicrobiaceae and Porphyromonadaceae. Others less well known in termite guts were Synergistaceae, Desulfobacteraceae, Rhodocyclaceae, Lachnospiraceae and Ruminococcaceae. Synergistaceae, Lachnospiraceae and Spirochaetaceae were well represented in the RNA data set, indicating a high-protein synthesis potential. Using 130,800 sequences from nine S. ruficeps DNA and RNA data sets, we estimated a high level of bacterial richness (4024 phylotypes at 3% genetic distance). Very few abundant phylotypes were site-specific; almost all (95%) abundant phylotypes, representing 97% of data set sequences, were detected in at least two S. ruficeps colonies. This study of a little-researched phylogenetically basal termite identifies core bacteria taxa. These findings will extend inventories of termite gut microbiota and contribute to the understanding of the specificity of termite gut microbiota.

Novosphingobium nitrogenifigens sp. nov., a polyhydroxyalkanoate-accumulating diazotroph isolated from a New Zealand pulp and paper wastewater.

A diazotroph capable of accumulating significant amounts of polyhydroxyalkanoate was isolated in New Zealand from a bioreactor treating nitrogen-deficient pulp and paper-mill effluent. Strain Y88T is Gram-negative, rod-shaped and positive for catalase, nitrate reductase and urease activities. The complete 16S rRNA gene sequence was most similar to those of other members of the genus Novosphingobium, the highest level of similarity (94.7%) being found with respect to the type strain of Novosphingobium stygium. The combined phenotypic, chemotaxonomic and sequence data show that while strain Y88T belongs to the genus Novosphingobium, it is distinct from all currently recognized Novosphingobium species. Therefore, strain Y88T represents the first nitrogen-fixing species of the genus Novosphingobium, for which the name Novosphingobium nitrogenifigens sp. nov. is proposed. The type strain is Y88T (=ICMP 16470T=DSM 19370T).