Soil associated with sea freight (shipping containers, flat racks and used machinery) arriving at New Zealand seaports was sampled for bacteria, fungi, nematodes, macro-invertebrates and plant seeds. Pseudomonads were selectively isolated, as several significant plant pathogens fall within this bacterial group. The mean and median sample weight collected from sea freight was found to be 417.3 and 152.7 g, respectively, with most recovered soil (73%) collected from the underside of shipping containers and flat rack containers. Likewise, for used machinery, most recovered soil (75%) was found under the machinery. Flat rack containers had significantly higher soil contamination compared to shipping containers and used machinery, but generally the counts and incidence of taxa were significantly lower compared to these other freight types. Viable bacteria, fungi, nematodes, seeds and arthropods were associated with the soil, with both counts g-1 and prevalence in samples varying with taxa, freight type, and location on the freight. Various regulated biosecurity organisms were recovered from the samples, including Aphelenchoides besseyi (rice white tip nematode), and seeds from genera such as Brachiaria, Cortaderia, Digitaria, Eragrostis and Sonchus. There were also live arthropod taxa that were not recorded as being present in New Zealand. No known plant pathogenic pseudomonads were identified through sequencing of the 16S ribosomal RNA gene. Shipping containers were found to be an important introduction pathway for exotic species, and therefore require careful monitoring and management. Comparisons of the incidence and mean number of organisms associated with soil on sea freight compared to a previous study with soil on footwear, generally showed that incidence and counts of many taxa were lower on sea freight, indicating that biosecurity risk can vary with pathway. However, prioritising one soil pathway over another according to the risks they present, and differentially allocating resources is problematic because the relative risk is dynamic, dictated by factors such as new pests or diseases entering the respective pathways.

Pastures represent about half of the global agricultural area and productivity losses from weeds are significant. The complex interactions between them and other pasture plants, livestock and the environment imply a need for innovative research that transforms pasture management. To this end, a horizon scan was conducted to identify relevant issues, questions, opportunities, and drivers. The drivers were ranked using three criteria: (1) is this a horizon (is the driver likely to become important in 10-20 years?); (2) will the research require stretchy science (is it currently not well addressed by the science community?); (3) is the research transformative (will successful scientific research in this area lead to significant changes to weed management in pastures?). We identified 11 major issues and 46 subordinate ones. The three highest ranked major issues were: (1) anticipated reductions in access to herbicides; (2) rethinking weed management under an ecosystem services paradigm; (3) responding to shifts in best practice and the regulations that are altering farm system planning to reduce farming's environmental impacts. We conclude that fundamental interdisciplinary research is needed that addresses biosecurity and weed management issues, while reducing the environmental footprint of farming and maintaining productivity.

AbstractIntroductionGlobally, soil‐borne plant pathogens are a significant contributor to plant yield loss in agricultural systems, including pasture production. The extent of soil biological constraints in high‐value dairy systems was assessed across three major dairying regions of New Zealand.Materials and MethodsRegional measures of disease pressure were obtained by comparing white clover, perennial ryegrass and plantain growth in 30 farm soils with (non‐pasteurised) or without (microwave pasteurised) their normal complement of microbial and nematode pathogens.ResultsPasteurising soils from the Waikato region led to significant average increases in clover (35%) and ryegrass (19%) shoot dry matter. Individual site yields increased up to 74% for clover and 38% for ryegrass. For Canterbury and Southland, there was no increase in plant yield with pasteurisation on a regional basis, but increases in either clover or ryegrass were significant at three individual farm sites. Across all regions, negative linear relationships between disease pressure (% growth change) and a proxy of soil organic matter quality (anaerobically mineralizable nitrogen:total nitrogen ratio were found for both clover and ryegrass, accounting for 34% of the variation in both clover and ryegrass growth. A positive linear relationship was found between disease pressure and Heterodera cyst nematode abundance, accounting for 33% and 17% of the variation in clover and ryegrass growth change, respectively. The most prevalent putative fungal and oomycete pathogens isolated from the roots and stem bases of clover and ryegrass seedlings were Fusarium oxysporum, Fusarium culmorum, and various Pythium species.ConclusionOur results suggest that root pathogens on New Zeland dairy farms are most prevalent and damaging in the Waikato region where economic modelling estimated clover and ryegrass root disease to cost 211 kg MS ha−1 year−1 in milk production and $909 ha−1 year−1 in farm profitability. Given the scale of these costs, targeted management of soil borne disease could present an economically viable approach to improving the resilience of these multi‐plant multi‐pathogen ecosystems.

Because plant and soil systems are strongly inter-linked, manipulating plant traits in intensively managed agricultural systems could be used to improve soil functioning and sustainability. However, we have little understanding of whether the impacts of plant traits on soil systems are modified by other management practices, such as fertiliser use. We tested whether relationships among plant traits, soil biota (microbes and nematodes) and soil functions (carbon and nitrogen (N) cycling) change with N fertiliser regime in a field-based, intensively managed experiment with high biomass removal. The experiment consisted of seven plant species compositions crossed with six N fertiliser rates (0-500 kg N ha(-1) year(-1)). Relationships among plant traits, soil biota and soil functions were often consistent across N fertiliser rates. However, the relationship of percentage N(2)fixer toqCO(2)shifted from negative to neutral as N rates increased, and the slope of several relationships of plant traits to N cycling indicators declined when >200 kg N ha(-1) year(-1)was added. The negative relationship of the fungal: bacterial ratio to N cycling indicators also became neutral when > 200 kg N ha(-1) year(-1)was added, and the relationships of bacterivore abundance and the plant parasitic index to respiration changed direction as N inputs increased. Some relationships of plant traits and soil biota to soil functions were in unexpected directions. This was sometimes associated with species-specific effects and inconsistent trait trade-offs within species. In general, conservative plant traits and fungal dominance were associated with low N cycling and cellulose paper decomposition rates, but high respiration rates. Nematode-based variables were better predictors of some functions than microbial ones; their incorporation into plant trait research could improve predictive power and system understanding. Synthesis and applications. Manipulating plant community traits can modify soil functions in intensively managed systems, but may result in larger changes where fertiliser nitrogen inputs are relatively low. Improved modelling that integrates how plant community traits, species-specific effects and management practices interact to determine soil functions will be required before managers can confidently predict the consequences of changing plant community traits.

• The soil microbiome refers to the diverse collection of microorganisms present in soil, many of which can mediate essential soil functions such as nutrient cycling and plant pest and disease suppression. • There is growing international interest in the potential to manipulate the soil microbiome to deliver improved agricultural and environmental outcomes. • Molecular techniques are rapidly increasing understanding of the soil microbiome structure and function but manipulating soil microbiomes for pasture resilience still presents significant science challenges.

We here report a study characterizing the potential for edible insects to act as a prebiotic by altering the bacterial composition of the human fecal microbiome, using batch cultures inoculated with fecal adult human donors. Black field cricket nymphs, grass grub larvae, and wax moth larvae were subjected to anin vitrodigestion to simulate the oral, gastric, and small intestinal stages of digestion. The digested material was then dialyzed to remove small molecules such as amino acids and free sugars to simulate removal of nutrients through upper gastrointestinal tract digestion. The retentate, representing the digestion resistant constituents, was then fermented in fecal batch cultures for 4, 7, and 15 h to represent rapid and longer fermentation times. Batch cultures without any added substrates were also set up to act as controls. Additionally, phosphate-buffered saline was used as a no-protein control and milk powder as "standard" protein control. At the end of the incubation period, the bacterial pellets were collected for microbiome analysis by 16S rRNA gene amplicon sequencing. Analysis of fecal cultures showed striking differences in community composition. Each substrate led to significant differences across a wide range of taxa compared to each other and PBS controls. Among the differences observed, digested grass grub larvae increased proportions ofFaecalibacteriumand thePrevotella2 group. Black field crickets increased the prevalence of theEscherichia-Shigellagroup,Dialistergenus, and a group of unclassifiedLachnospiraceae. Wax moth larvae promoted the expansion of the same group of unclassifiedLachnospiraceaeand theEscherichia/Shigellagroup. The increasedFaecalibacteriumobserved in the cultures with grass grub larvae represents a noteworthy finding as this bacterium is widely thought to be beneficial in nature, with demonstrated anti-inflammatory properties and associations with gut health. We conclude that insects can differentially modulate the microbiome composition in batch cultures inoculated with adult fecal material after simulatedin vitrodigestion. Although the physiological impactin vivoremains to be determined, this study provides sound scientific evidence that investigating the potential for consuming insects for gut health is warranted.

The invertebrate pests most commonly affecting New Zealand's pastoral-based production in 'average' years cause losses of between $1.7B and $2.3B p.a. of which up to $0.9B occur on sheep and beef farms and $1.4B on dairy farms. The native scarab grass grub is the most costly pest causing losses of $140-380 M on dairy farms and $75-205 M on sheep and beef farms annually. The exotic scarab, black beetle, although only affecting approximately 1 M ha, costs dairy farmers up to $223 M and sheep and beef farmers up to $19 M annually. Porina cause losses up to $84 M and $88 M respectively. Pasture nematodes are estimated to cost up to $274 M p.a. for dairy farmers and $326 M p.a. for sheep and beef farmers. Two exotic pests, Argentine stem weevil (ASW) and clover root weevil (CRW) are causing damage estimated at up to $200 M p.a. and $235 M p.a. respectively in dairy and sheep and beef pastures. While CRW is subject to successful biological control management it still causes considerable losses. Lesser pests also contribute to lost production, particularly as they often coexist with more major pests. However, their economic cost to New Zealand is difficult to calculate due to the variable nature of infestations on both temporal and spatial scales. At farm and paddock level, it is abundantly clear that substantial savings could be made if pest management is achieved. It is equally clear that in many instances the tools to do so are limited but if developed would contribute substantially to farm profitability.

Effective surveillance for early detection of invasive alien species in natural ecosystems, or on valued plants found in modified areas, could prevent potentially devastating and costly impacts (whether environmental, economic or cultural) of new invasions on the invaded country. Surveillance technologies are often constrained by a range of factors. Determining which species present a significant risk before they reach the border is an effective strategy to minimize the possibility of invasion and/or the impact of invasion. Surveillance of sentinel plants provides an important tool to strengthen biosecurity programs assisting with i) detecting and identifying insect pests, nematodes and plant diseases that could potentially invade uncolonized countries, and ii) developing pest risk analysis profiles to eliminate or mitigate the risk of arrival. This review examines some of the challenges and opportunities provided by sentinel plant research and discusses the factors that could affect the success of their use for biosecurity risk assessment and surveillance in the New Zealand context.

The sensitivity of four radish (Raphanus sativus L.) cultivars, Baladey “B,” Cherry Belle “CB,” Prinz Rotin “PR,” and Scarlet Globe “SG” to 80 nL L−1 ozone is assessed in fumigation chambers. O3 visible injury symptoms appears as brown spots to chlorotic areas, but the response is cultivar‐specific. O3 induces higher content of H2O2 and lipid peroxidation (it is estimated as malondialdehyde (MDA) content), leading to drastic visible injury symptoms in B compared to the other cultivars. Root and shoot dry weights, Chlorophyll a, net photosynthetic rates, and chlorophyll fluorescence are reduced in B to a greater extent than the other cultivars. On the other hand, stomatal conductance increased in B and CB (+52 and +24%, respectively) due to O3‐exposure, while it was decreased by 35% in PR and SG. Exposure to O3 generates oxidative stress leading to stimulation of antioxidative defense systems. SG shows the highest induction of catalase (CAT), superoxide dismutase (SOD), and glutathione reductase (GR), while B had the lowest activities of these antioxidative enzymes, but had the highest H2O2 and MDA content. The higher accumulation of H2O2, poor induction of antioxidative enzymes, and increased stomatal conductance led to severe visible injury and drastic inhibition in photosynthetic rates and growth in B than other cultivars depicting its higher sensitivity toward O3. Therefore, it can be used as a bioindicator for O3 pollution worldwide.

This experiment, implemented at a Southland site representative of the lower South Island, New Zealand, and monitored for three full years from 2012/2013 to 2015/2016, was one of four throughout New Zealand that investigated the yield and nutritive value rankings of perennial ryegrass cultivars sown with or without white clover and with high (225kgN/ha/annum) or low (50kgN/ha/annum) application rates of nitrogen fertiliser. High inputs of nitrogen (N) fertiliser increased annual total dry matter (DM) yields and seasonal yield in 12 out of 18 measurement occasions. Adding white clover increased both annual dry matter yield and seasonal yield in 7 out of 18 measurement occasions. An interaction between clover and N input occurred on 10 occasions. Over time, the low N minus clover treatment became progressively lower yielding compared with the other three treatments. Ryegrass cultivar by clover interactions occurred in the latter half of the experiment. When analysed using ryegrass characteristics, early- and late-heading ryegrass cultivars produced similar yields when sown without clover, but late-heading ryegrass cultivars had a higher yield than early-heading pastures when sown with white clover (P<.001). Tetraploid ryegrasses had lower yields than diploid ryegrasses when sown without clover (P=.001). This difference was reduced when white clover was included. There was no significant interaction with ryegrass morphology when sown with white clover, as dense ryegrass cultivars always had a higher yield than open ryegrass cultivars (P=.005). Interactions occurred between N input levels and ryegrass cultivar. These occurred in early spring or late spring of each year. Analysis by ryegrass characteristics identified consistent interactions. Ryegrass cultivars with a late-heading date were higher yielding than early-heading cultivars at low N inputs (P=.029), but they were not different at high N inputs. Diploid and tetraploid ryegrass cultivars were similar with low N inputs, but diploid cultivars out-yielded tetraploid cultivars at high N inputs (P=.005). Significant interactions between N inputs and white clover addition affected herbage nutritive value but this did not extend to consistent interactions with ryegrass cultivar. Invertebrate pest numbers were generally low and unlikely to have influenced clover/ryegrass interactions or cultivar ranking. Results from this site suggest that ranking of ryegrass cultivars needs to account for both the inclusion of clover and variation in N fertiliser input.

Poor persistence of perennial ryegrass swards is a common problem; however, there is a lack of long-term studies to understand the mechanisms associated with poor persistence. This study describes an experiment to test the hypothesis that high ryegrass seeding rates (>18 kg seed per ha) reduce long-term population persistence because of smaller plant size and poorer survival during the first year after sowing. Four cultivars, representing four functional types of perennial ryegrass, were sown at five seeding rates (equivalent to 6, 12, 18, 24 and 30 kg seed per ha) with white clover in three regions of New Zealand. Swards were monitored for 5 years. No evidence was found to indicate a lack of persistence of ryegrass-based swards sown at higher seeding rates. During the first year, swards sown at higher seeding rates had greater herbage accumulation (except at the Waikato site), greater ryegrass tiller density and greater ryegrass content. This initial impact of high seeding rates had largely dissipated by the fourth year, resulting in swards with similar annual herbage accumulation, tiller density and botanical composition. Similarly, there were relatively few differences among cultivars for these variables. Although high seeding rates did not negatively impact sward persistence, geographical location did, with strong evidence of ryegrass population decline at the Waikato site for all treatment combinations, some decline in Northland, and stable populations in Canterbury. It is possible that productive perennial ryegrass pastures can only be sustained for 4-5 years in some situations, even when the best ryegrass technology and management practices are used.

The implementation and monitoring of the treatments in the core experiment (eight perennial ryegrass cultivars grown under four combinations of plus/minus clover and high/low nitrogen (N)) at a dryland Waikato site is described. The N x clover interaction was significant in 10 of the 17 seasonal or total annual herbage accumulation (HA) data sets available, caused by substantially lower annual HA in the low N minus clover treatment compared with all other treatments. Five significant scaling-type clover x cultivar interactions in HA were evident. In all cases, HA was greater in plus clover than minus clover treatments, but more so for some cultivars than others. However, the interactions were not consistent across seasons, and different cultivars or phenotypic contrast groups were involved in many cases. The hypothesis that relative HA rankings of ryegrass cultivars do not differ when ryegrass is grown in monoculture versus mixtures with white clover is supported.

This paper describes how the core treatments in the national experimental design (all combinations of with/without clover and high/low nitrogen (N)) were implemented at an irrigated Canterbury site and monitored for three years. There was a consistent N x clover interaction, caused mainly by substantially lower yields in the Low N without clover treatment compared with all other treatments. At the phenotypic contrast level, the main effect of perennial ryegrass heading date on white clover content was significant in four of the nine seasonal data sets available: on these occasions, mixtures based on mid-season heading cultivars had higher clover content than later season-heading cultivars. However, this difference was not sufficient to cause any cloverxcultivar interactions in herbage accumulation. Hence, the hypothesis that relative yield rankings of ryegrass cultivars do not differ when ryegrass is grown in monoculture or in mixture with white clover is supported.

The implementation and monitoring of the treatments in the core experiment (eight perennial ryegrass cultivars grown under four combinations of plus/minus clover and high/low nitrogen (N)) at a dryland Manawatu site are described. Herbage accumulation (HA) was significantly greater for high N than for low N in 14 out of 21 annual and seasonal totals. There were no significant main effects of clover and there were no interactions between clover and N level. With the exception of autumn 2016, there were significant differences among ryegrass cultivars in all seasons, but no clover × cultivar or N × cultivar interactions for HA or nutritive value (NV). With the caveat that successive droughts reduced the proportion of white clover to low levels, the hypothesis that relative HA or NV rankings of ryegrass cultivars do not differ when ryegrass is grown in monoculture or in mixtures with white clover is supported.

Meloidogyne minor Karssen et al. 2004 was collected from perennial ryegrass (Lolium perenne L.) growing in a sports ground in Christchurch, New Zealand. This is a new record for M. minor, the first report of this nematode occurring in New Zealand, and the second report from the southern hemisphere (after Chile). In general, the New Zealand isolate of M. minor corresponds well to the descriptions of M. minor given by Karssen et al. (2004). The New Zealand isolate is characterized by having a female with dorsally curved stylet, 13-14 μm long, with transversely ovoid knobs slightly sloping backwards from shaft; rounded perineal pattern; and male with stylet 16-19 μm long, large transversely ovoid knobs sloping slightly backwards from shaft; head region not set off, labial disc elevated, lateral lips prominent; and second stage juvenile 370-390 μm long, with hemizonid posterior but adjacent to excretory pore; tail 53-63 μm long; and a distinct hyaline tail terminus 14-18 μm long. In addition, molecular phylogeny using near full length small subunit (SSU), D2/D3 expansion segments of the large subunit (LSU), the internal transcribed spacer region (ITS1 and 2), and the intergenic spacer (IGS2) of the ribosomal rDNA supports the identification.

Plant parasitic nematodes (PPN) are known to survive periods of desiccation, an ability that increases the risk of them surviving unintentional transport between countries. To investigate nematode survival in soil subject to prolonged storage, soil collected from a native forest and an organic orchard was stored separately in cupboards at ambient temperature for 36 months. Subsamples were taken at 0, 3, 6, 12, 13, 24 and 36 months to determine the presence of plant parasitic and total nematodes using a standard misting technique. Pratylenchus was used as a model to determine if PPNs that had been under prolonged storage were able to infect plant hosts at 13, 24 and 36 months. Overall, the total number of nematodes recovered from stored soil declined over time, with differences in species diversity determined by molecular methods, related to soil origin. No PPN were recovered in soil stored beyond 13 months using the three-day misting technique. By comparison, Pratylenchus nematodes, using a baiting method, were found to successfully invade host plant roots (ryegrass and white clover) even after 36 months storage and were observed to produce offspring at 13 months. Baiting was not effective for Pratylenchus found in soil originally collected from the forest but was for orchard soil, a result attributed to the lack of suitable host plants for the Pratylenchus species found in forest soil. This study demonstrated, that in protected environments, nematodes could survive for at least 36 months and were observed to produce offspring at 13 months. Baiting with a host plant was more sensitive in detecting nematodes than using the misting extraction technique, although this approach only works where the host plant is known. Without a priori knowledge of the nematode-plant host association, plant baiting may also produce false negatives. In the context of plant biosecurity and providing an accurate risk assessment in soil contaminants, the development of a generic test for PPN that induces nematodes in a resting stage to emerge and respond to a cue would enhance the probability of detection. However, as assessments at the border are often time limited, a molecular based bioassay that can be used to indicate the presence of multiple species of live PPN species may be a more feasible option for risk assessments.

Soil frequently occurs as a contaminant on numerous sea, land and air transport pathways. It can carry unwanted invasive species, is widely recognized as a biosecurity risk, and is usually strictly regulated by biosecurity authorities. However, little is known about relative risk levels between pathways, thus authorities have limited capability to identify and target the riskiest soil pathways for management. We conducted a an experiment to test the hypotheses that biosecurity risks from soil organisms will increase both with declining transport duration and with increasing protection from environmental extremes. Soil was collected from two sites, a native forest remnant and an orchard, and stored on, in and under sea containers, or in cupboards, and assayed after 0, 3, 6 and 12 months for bacteria, fungi, nematodes and seeds. Results showed that viability of Pseudomonas spp., bacteria, nematodes and plants declined over 12 months, irrespective of soil source. Also, mortality of most biota was higher when exposed to sunlight, moisture and desiccation than when protected. However, bacterial and fungal numbers were higher in exposed environments, possibly due to ongoing colonization of exposed soil by airborne propagules. The results were consistent with our observations of organisms in soil intercepted from airports and sea ports, and indicated there is potential to rank risks from transported soils based partly on transport duration and environmental exposure. This would help authorities to optimally allocate management resources according to pathway-specific risks.

Abstract Herbage accumulation, botanical composition, tiller density and insect pest populations were monitored over 6 years for four perennial ryegrass functional types grazed by dairy cows in the Waikato. The aim was to identify genotypic and environmental factors contributing to ryegrass persistence failure in the upper North Island. Perennial ryegrass content of pastures declined as low as 60% of total herbage mass (HM) in summer of the first 3 years but recovered in autumn to at least 75%. However, following two successive severe summer/autumn droughts in Years 2 and 3, and subsequent grass grub larvae populations exceeding the damage threshold of 200/m2, by 4 years after sowing (autumn 2015), ryegrass had fallen to 20% of HM. This pattern was repeated in the following 2 years, and was not prevented by any combination of ryegrass functional type, endophyte, seeding rate, or best-practice dairy cattle grazing and soil nutrient management. The abiotic and biotic environmental stresses dominated all other factors. Keywords: pasture persistence, perennial ryegrass, drought, insects, black beetle, dairy systems

White clover (Trifolium repens) is the key legume component of New Zealand pastoral agriculture due to the high quality feed and nitrogen inputs it provides. Invertebrate pests constrain white clover growth and this study investigated rhizosphere-associated fungal controls for two of these pests and attempts to disentangle the underpinning mechanisms. The degree of suppressiveness of 10 soils, in a latitudinal gradient down New Zealand, to added Meloidogyne hapla and Costelytra zealandica scarab larvae was measured in untreated soil. Most of the soils showed no suppressive activity against these pests but two showed activity against M. hapla and two against C. zealandica. Rhizosphere fungi responsible for pest suppressive responses were elucidated via next-generation sequencing. In the M. hapla-suppressive soils nematode-trapping Orbiliomycetes fungi were present in significantly greater abundance than non-suppressive soils and their abundance increased further with addition of M. hapla. A comparison of plant growth and the rhizosphere fungal community between untreated and irradiated soil was carried out on 5 of the 10 soils using Pyronota as the scarab larvae. Soil irradiation either: reduced (by 60-70%); increased (16×) or made no difference to white clover growth across the five soils tested, illustrating the range of microbial impacts on plant production. In one of the M. hapla suppressive soils irradiation resulted in a significant increase in nematode galling suggesting that Orbiliomycetes fungi were indeed responsible for the suppressive effect. Lack of consistent changes in soil macronutrients and pH post-irradiation suggest these were not responsible for plant or invertebrate responses. The use of next generation sequencing in controlled pot trials has allowed identification of a potential biological control organism and bioindicator for M. hapla suppression.