Interactions between 14C-labelled atrazine and the soil microbial biomass in relation to herbicide degradation

A laboratory incubation experiment was set up to determine the effects of atrazine herbicide on the size and activity of the soil microbial biomass. This experiment was of a factorial design (0, 5, and 50 g g^–1 soil of non-labelled atrazine and 6.6×10³ Bq g^–1 soil of ¹⁴C-labelled atrazine) x (0, 20, and 100 g g^–1 soil of urea-N) x (pasture or arable soil with a previous history of atrazine application). Microbial biomass, measured by substrate-induced respiration and the fumigation-incubation method, basal respiration, incorporation of ¹⁴C into the microbial biomass, degradation of atrazine, and ¹⁴C remaining in soil were monitored over 81 days. The amount of microbial biomass was unaffected by atrazine although atrazine caused a significant enhancement of CO₂ release in the non-fumigated controls. Generally, the amounts of atrazine incorporated into the microbial biomass were negligible, indicating that microbial incorporation of C from atrazine is not an important mechanism of herbicide breakdown. Depending on the type of soil and the rate of atrazine application, 18–65% of atrazine was degraded by the end of the experiment. Although the pasture soil had twice the amount of microbial biomass as the arable soil, and the addition of urea approximately doubled the microbial biomass, this did not significantly enhance the degradation of atrazine. This suggests that degradation of atrazine is largely independent of the size of the microbial biomass and suggests that other factors (e.g., solubility, chemical hydrolysis) regulate atrazine breakdown. A separate experiment conducted to determine total amounts of ¹⁴C-labelled atrazine converted into CO₂ by pasture and arable soils showed that less than 25% of the added ¹⁴C-labelled atrazine was oxidised to ¹⁴CO₂ during a 15-week period. The rate of degradation was significantly greater in the arable soil at 24%, compared to 18% in the pasture soil. This indicates that soil microbes with previous exposure to atrazine can degrade the applied atrazine at a faster rate.     

Prediction of field atrazine persistence in an allophanic soil with Opus2

A modified version of the model Opus was applied to measurements of soil water dynamics and atrazine (6-chloro-N2-ethyl-N4-isopropyl-1,3,5-triazine-2,4-diamine) persistence in a Bruntwood silt loam soil (Haplic Andosol, FAO system) in Hamilton, New Zealand. The modified model, Opus2, is briefly described and parameter estimation for the simulations is discussed. Soil water dynamics were more accurately described by applying measured soil hydraulic properties than by estimating them using pedotransfer functions. A parameter sensitivity analysis revealed that degradation was the most relevant process in simulating pesticide behaviour by Opus2. The Arrhenius equation incorporated in Opus2 did not correctly describe the effect of temperature on degradation rates obtained at 10, 20 and 30 degrees C. However, as the Arrhenius coefficient is a very sensitive parameter and soil temperature variation was relatively narrow in the field, the Arrhenius coefficient was approximated from the laboratory study. The simulation results obtained were superior to modelling at constant temperature. Field measured persistence of atrazine in the topsoil was underpredicted using the half-life determined in the laboratory at 10 degrees C. Modelling with a lag phase followed by accelerated degradation by use of a sigmoidal degradation equation in Opus2 significantly improved the modelling results. Nevertheless, degradation processes in the laboratory under controlled conditions did not accurately represent field dissipation, however well the laboratory degradation data could be described by simple kinetic equations. The study indicates the importance of improving field techniques for measuring degradation, and developing laboratory protocols that yield degradation data that are more representative of pesticide dynamics in field soils.     

DNA vaccine encoding avian influenza virus H5 and Esat-6 of Mycobacterium tuberculosis improved antibody responses against AIV in chickens

The H5 gene of avian influenza virus (AIV) strain A/chicken/Malaysia/5744/2004(H5N1) was cloned into pcDNA3.1 vector, and Esat-6 gene of Mycobacterium tuberculosis was fused into downstream of the H5 gene as a genetic adjuvant for DNA vaccine candidates. The antibody level against AIV was measured using enzyme-linked immunosorbent assay (ELISA) and haemagglutination inhibition (HI) test. Sera obtained from specific-pathogen-free chickens immunized with pcDNA3.1/H5 and pcDNA3.1/H5/Esat-6 demonstrated antibody responses as early as 2 weeks after the first immunization. Furthermore, the overall HI antibody titer in chickens immunized with pcDNA3.1/H5/Esat-6 was higher compared to the chickens immunized with pcDNA3.1/H5 (p < 0.05). The results suggested that Esat-6 gene of M. tuberculosis is a potential genetic adjuvant for the development of effective H5 DNA vaccine in chickens.     

Mapping QTLs using a novel source of salinity tolerance from Hasawi and their interaction with environments in rice

Background

Salinity is one of the most severe and widespread abiotic stresses that affect rice production. The identification of major-effect quantitative trait loci (QTLs) for traits related to salinity tolerance and understanding of QTL × environment interactions (QEIs) can help in more precise and faster development of salinity-tolerant rice varieties through marker-assisted breeding. Recombinant inbred lines (RILs) derived from IR29/Hasawi (a novel source of salinity) were screened for salinity tolerance in the IRRI phytotron in the Philippines (E1) and in two other diverse environments in Senegal (E2) and Tanzania (E3). QTLs were mapped for traits related to salinity tolerance at the seedling stage.

Results

The RILs were genotyped using 194 polymorphic SNPs (single nucleotide polymorphisms). After removing segregation distortion markers (SDM), a total of 145 and 135 SNPs were used to construct a genetic linkage map with a length of 1655 and 1662 cM, with an average marker density of 11.4 cM in E1 and 12.3 cM in E2 and E3, respectively. A total of 34 QTLs were identified on 10 chromosomes for five traits using ICIM-ADD and segregation distortion locus (SDL) mapping (IM-ADD) under salinity stress across environments. Eight major genomic regions on chromosome 1 between 170 and 175 cM (qSES1.3, qSES1.4, qSL1.2, qSL1.3, qRL1.1, qRL1.2, qFWsht1.2, qDWsht1.2), chromosome 4 at 32 cM (qSES4.1, qFWsht4.2, qDWsht4.2), chromosome 6 at 115 cM (qFWsht6.1, qDWsht6.1), chromosome 8 at 105 cM (qFWsht8.1, qDWsht8.1), and chromosome 12 at 78 cM (qFWsht12.1, qDWsht12.1) have co-localized QTLs for the multiple traits that might be governing seedling stage salinity tolerance through multiple traits in different phenotyping environments, thus suggesting these as hot spots for tolerance of salinity. Forty-nine and 30 significant pair-wise epistatic interactions were detected between QTL-linked and QTL-unlinked regions using single-environment and multi-environment analyses.

Conclusions

The identification of genomic regions for salinity tolerance in the RILs showed that Hasawi possesses alleles that are novel for salinity tolerance. The common regions for the multiple QTLs across environments as co-localized regions on chromosomes 1, 4, 6, 8, and 12 could be due to linkage or pleiotropic effect, which might be helpful for multiple QTL introgression for marker-assisted breeding programs to improve the salinity tolerance of adaptive and popular but otherwise salinity-sensitive rice varieties.

     

Variability in concentrations of zinc in serum and feed when using zinc oxide as a supplement for the prevention of facial eczema

AIMS: To determine the variability of concentrations of Zn in feed, when used as a supplement to prevent facial eczema, and to determine the variability in concentrations of Zn in serum between cows and herds that are being supplemented with ZnO in feed, using in-shed feeders or on a feed pad.

METHODS: Sixteen commercial dairy farms in the Waikato region of New Zealand were enrolled, that were supplementing cows with ZnO in the feed using either an automatic in-shed feeder (ASF) or a feed pad (FP) using a feed-out or mixer wagon. On each farm 10 cows were selected by the farmer, that were assumed to be representative of the age and liveweight of the herd. Four hours after supplement feeding, each cow was weighed and a blood sample collected for measurement of concentrations of Zn in serum. Three samples of feed were collected from each farm for Zn analysis, from the beginning, middle and end of the feed being distributed. Levene’s test for homoscedasticity was used to analyse whether there were differences in variation of individual concentrations of Zn in serum, and in the feed, between the two feeding systems. Multivariable linear regression was used to examine associations between age, feeding method or liveweight and concentrations of Zn in serum, after accounting for the variability between farms.

RESULTS: Of the 163 cows sampled, concentrations of Zn in serum were between 20–35?µmol/L in 75/163 (46 (95% CI=38–54)%) cows; were <20?µmol/L in 71/163 (44 (95% CI=36–52)%) cows, and >35?µmol/L in 17/163 (10 (95% CI=6–16)%) cows. The variation in concentrations of Zn in serum in individual cows differed between farms (p<0.001), and the variability was greater for cows fed using a FP than ASF (p<0.001). There was no difference in the variation of concentrations of Zn in feed between the two feeding methods (p=0.54), but concentrations of Zn in serum were associated with the amount of Zn offered in feed (p=0.008).

CONCLUSIONS AND CLINICIAL RELEVENCE: There was significant variability between farms in the concentrations of Zn in the serum of cows being supplemented with ZnO in feed. Only 46% of cows sampled had concentrations of Zn between 20–35?µmol/L. Effective management of facial eczema should include monitoring Zn in the feed and in serum to ensure cows are receiving the correct dose they require.     

A high virulence and pathotype diversity of Puccinia striiformis f.sp. tritici at its centre of diversity,the Himalayan region of Pakistan

Information on the pathogen virulence profile and diversity across locations is crucial for host germplasm improvement and deployment. The rapid acquisition of virulence to host resistance by the wheat yellow/stripe rust pathogen (Puccinia striiformis f.sp. tritici: PST), makes it crucial to know about its virulence and pathotype diversity. Recent studies have shown the plausible centre of origin of the pathogen in the Himalayan region, with Pakistan being the most ancestral to all other worldwide populations. To assess the status of virulence and pathotype diversity in the Himalayan region of Pakistan, a set of 127 PST infected wheat samples from eight locations were collected, multiplied and pathotyped using a set of 36 differential lines from the world set, European and Chinese sets, and 9 Avocet Yr isolines. Virulence (Vr) was recorded to 18 out of 24 tested yellow rust resistance (Yr) genes, while a total of 53 pathotypes were detected out of 127 isolates tested. Virulence was found to the resistance genes rarely deployed in Pakistan (Vr8) or even worldwide level (Vr5), while virulence to Vilmorin 23 (Yr3+) was absent in Pakistan, which is common in Europe. None of the pathotypes was dominant across all locations, however, no clear spatial structuring was observed for the studied locations. Our results suggested a high virulence and pathotype diversity in line with the previously proposed potential role of sexual recombination in the temporal maintenance of PST in the Himalayan region of Pakistan. This information should be useful in host resistance gene improvement and deployment.     

Co‐infection by Botryosphaeriaceae and Ilyonectria spp. fungi during propagation causes decline of young grafted grapevines

Decline of newly planted, grafted grapevines is a serious viticultural problem worldwide. In the Riverina (New South Wales, Australia), characteristic symptoms include low fruit yields, very short shoots and severely stunted roots with black, sunken, necrotic lesions. To determine the cause, roots and wood tissue from affected plants in 20 vineyards (Vitis vinifera cv. Chardonnay grafted to V. champini cv. Ramsey rootstock) were assayed for microbial pathogens. Ilyonectria spp. (I. macrodidyma or I. liriodendra, producers of phytotoxin brefeldin A, BFA, and cause of black foot disease of grapevines) and Botryosphaeriaceae spp. (predominantly Diplodia seriata) were isolated from rootstocks of 100 and 95% of the plants, respectively. Togninia minima and Phaeomoniella chlamydospora (cause of grapevine Petri disease) were isolated from 13 and 7% of affected plants, respectively. All Ramsey rootstock stems of grafted plants sampled from a supplier nursery were infected with Ilyonectria spp. and D. seriata. Diplodia seriata, but not Ilyonectria spp., was also isolated from 25% of canes sampled from the rootstock source block. Root inoculation of potted, disease‐free Chardonnay plants with Ilyonectria isolates from diseased vineyards caused typical disease symptoms, while co‐inoculation with Botryosphaeriaceae spp. increased disease severity. This is the first study to show that a major cause of young grapevine decline can be sequential infection by Botryosphaeriaceae from rootstock cuttings and Ilyonectria spp. from nursery soil. Although the Petri disease fungi were less common in young declining grafted grapevines in the Riverina, they are likely to contribute to the decline of surviving plants as they mature.     

Effects of Ufra Disease on Yield Loss of Deepwater Rice in Bangladesh

Abstract

Ufra, caused by the stem nematode, Ditylenchus angustus (Butler, 1913) Filipjev, 1936, is a serious disease of deepwater rice in southern Bangladesh. It was studied between 1977 and 1979 as part of a joint project with the Overseas Development Administration at the Bangladesh Rice Research Institute. Three symptom types are distinguished according to the extent of panicle emergence: Ufra1 (none), Ufra2 (partial) and Ufra3 (complete). All three represent total loss from that panicle. The number of Ufra2 as a percentage of the total number of panicles present in a specified area is selected as a disease index (UfraII). A function relating yield to the level of UfraII is proposed on the basis of selective samples from a field with a well-defined ufra patch. This function is compared with another derived from random samples in different fields along a linear transect through an area where ufra is endemic. Loss of panicle density, i.e. panicles/m², is identified as a major component of yield loss, approximately equal to the loss associated with all three symptom types added together. Although severe and widely distributed throughout the southern part of the deepwater rice region in 1977 and 1978, it was very difficult to find in 1979 when there was a spring drought. Ufra is worse in wet years, in areas which flood early, and where the crop is harvested late. The length of the over-winter decay phase appears to be a critical factor regulating the survival of the pathogen from one season to the next. This suggests an approach for ufra control.     

Two new species,Phytophthora nagaii sp. nov. and P. fragariaefolia sp. nov., causing serious diseases on rose and strawberry plants,respectively, in Japan

A new disease of rose was noticed in Chiba Prefecture of Japan in 1968, and the pathogen was initially identified as Phytophthora megasperma based on morphological characteristics. Similar Phytophthora isolates have since been collected from rose plants in Chiba, Kanagawa, and Shizuoka Prefectures. In 2005, several Phytophthora isolates were recovered from crowns of strawberry plants in Hokkaido Prefecture. These were considered to be members of a new species. In this study, we re-examined all these isolates using morphological and physiological studies and a multilocus phylogenetic analysis. The rose and strawberry isolates were mostly similar morphologically and physiologically, with some exceptions. The rose isolates differed significantly from P. megasperma sensu stricto and other related Phytophthora species. The rose and strawberry isolates had external proliferation of sporangia, characteristic funnel-shaped oogonia, predominantly paragynous antheridia, and fast growth rates of 10.5 mm/24 h at an optimum temperature of 28 °C. In the multilocus phylogenetic tree constructed using sequences from the rDNA ITS regions, rDNA LSU, and the translation elongation factor 1-α, β-tubulin and coxI genes, they formed a distinct monophyletic group in clade 7 with strong bootstrap support. The rose and strawberry isolates separated into two distinct groups. The results indicate that the rose and strawberry isolates constitute two separate species, designated here as Phytophthora nagaii and P. fragariaefolia.