Agroecological characterization of Xiphinema index in Spain1

The agroecological characteristics of Xiphinema index were determined by studying its spatial and temporal distribution in different environmental conditions in Spain. The host range, life cycle in the Mediterranean continental climate and its relationships with grapevine fanleaf nepovirus are also reported. X. index is widespread in grape-growing areas in a wide range of soil textures and pH, even at high percentage of carbonates. However, it seems to prefer sandy-loam and sandy-clay loam soils, and populations decrease when soil carbonates increase. Its spatial distribution is random but it has a tendency to be found in the deepest clay horizons, where soil moisture is retained. Availability of moisture seems to be the limiting factor for nematode development. Grapevine and fig are the main host plants but X. index is also associated with fruit trees, ornamental crops, vegetable crops, woodlands and other uncultivated soils. Its life cycle in central Spain, where temperature ranges from <0°C until March to above 40°C from May—June until September, takes 6–8 weeks to complete. X. index was found in 14% of all vineyards sampled and in 50% of the fanleaf virus-infected vineyards. Transmission tests using bait plants were successful and the detection of the virus by means of ELISA test in a mimimun of five nematodes was achieved. Fanleaf-like symptoms often occurred in the absence of the virus, as demonstrated by ELISA.     

Cytochrome c oxidase subunit 1 analysis of <Emphasis Type="Italic">Xiphinema diversicaudatum,X. pachtaicum,X. simile</Emphasis> and <Emphasis Type="Italic">X. vuittenezi</Emphasis> (Nematoda,Dorylaimida)

Genetic analyses using sequences of partial cytochrome c oxidase subunit 1 (cox1) gene of mitochondrial DNA were conducted to determine the extent of genetic variation within and among Xiphinema diversicaudatum, X. pachtaicum, X. simile and X. vuittenezi populations. Pairwise distance among the four species was 22.5 to 31.2%. Four different sequence variants of cox1 were determined among six populations of X. diversicaudatum and three variants among three populations of X. simile. Nucleotide variation was detected at 18 of 414 bp (1.9 to 2.7%) in X. diversicaudatum and 4 of 435 bp (0.2 to 0.9%) in X. simile. All changes were at silent sites. No nucleotide variation was detected within three populations of X. pachtaicum and within three populations of X. vuittenezi.     

Development of a real‐time PCR method for the detection of the dagger nematodes Xiphinema index,X. diversicaudatum,X. vuittenezi and X. italiae,and for the quantification of X. index numbers

The ectoparasitic dagger nematodes Xiphinema index and Xiphinema diversicaudatum, often at low numbers in the soil, are vectors of grapevine nepoviruses, which cause huge agronomical problems for the vineyard industry. This study reports a method, based on real‐time PCR, for the specific detection of these species and of the closely related non‐vector species Xiphinema vuittenezi and Xiphinema italiae. Specific primers and TaqMan probes were designed from the ribosomal DNA internal transcribed spacer 1 (ITS1), enabling the specific detection of single individuals of each of the X. index, X. diversicaudatum, X. italiae and X. vuittenezi species whatever the nematode population. The specificity of detection and absence of false positive reaction were confirmed in samples of each species mixed with the three other Xiphinema species or mixed with nematodes representative from other genera (non‐plant‐parasitic Dorylaimida, Longidorus sp., Meloidogyne spp., Globodera spp. and Pratylenchus sp.). The method was shown to be valid for the relative quantification of X. index numbers through its use, from crude nematode extracts of soil samples, in a greenhouse assay of grapevine accessions ranging from highly susceptible to resistant. As an alternative to time‐consuming microscopic identification and counting, this real‐time PCR method will provide a fast, sensitive and reliable diagnostic and relative quantification technique for X. index nematodes extracted from fields or controlled conditions.     

Prevalence, polyphasic identification, and molecular phylogeny of dagger and needle nematodes infesting vineyards in southern Spain

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of grapevine plants in southern Spain were investigated. Nematode surveys were conducted on 77 vineyards during the spring seasons of 2006, 2007 and 2008 in the main Andalusian grapevine-growing areas, including the provinces of Cádiz, Córdoba, and Huelva. Morphological and morphometrical studies identified two Longidorus and nine Xiphinema species, viz.: Longidorus alvegus, L. magnus, Xiphinema adenohystherum, X. hispidum, X. index, X. italiae, X. lupini, X. nuragicum, X. pachtaicum, X. rivesi, and X. turcicum. Overall, frequencies of infestation were, in decreasing order: X. pachtaicum 90.8%, X. index 30.3%, X. italiae 13.2%, L. magnus 11.8%, X. hispidum 7.9%, X. lupini 3.9%, L. alvegus and X. rivesi 2.6%, and X. adenohystherum, X. nuragicum and X. turcicum 1.3%. Xiphinema hispidum, X. lupini, L. alvegus and L. magnus were compared with nematode type specimens and are reported for the first time in Spain. Furthermore, the male of L. alvegus is described for the first time in the literature. Molecular characterisation of these species using D2–D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids. The monophily of the genera Xiphinema and Longidorus was accepted and the genera Paralongidorus and Xiphidorus were rejected by the Shimodaira-Hasegawa test based on tree topologies.     

Validation of the specificity and sensitivity of species-specific primers that provide a reliable molecular diagnostic for <Emphasis Type="Italic">Xiphinema diversicaudatum</Emphasis>, <Emphasis Type="Italic">X. index</Emphasis> and <Emphasis Type="Italic">X. vuittenezi</Emphasis>

Xiphinema diversicaudatum and X. index are vector nematode species of economic importance in viticulture regions as they can transmit Arabis Mosaic, Grapevine Fanleaf and Strawberry Latent Ringspot viruses to grapevine. Wang et al. (2003) designed species-specific diagnostic primers from ribosomal genes for both these vector species as well as a vector and a non-vector species X. italiae and X. vuittenezi, respectively. Our study aimed to confirm the specificity and determine the sensitivity and reliability of the primers for the two vector species, X. diversicaudatumand X. indexwhen challenged with closely related longidorid species and general nematode communities typical of vineyard soil. With one exception, no PCR product was observed when the primers were tested against six Longidorus, one Paralongidorus and one Xiphinema non-target species. Occasionally (three out of eight replicate PCR reactions) a weak PCR product was noted when primers for X. index were tested with L. elongatus. Furthermore, when challenged with a range of non-target nematode species comprising the nematode community typical of viticulture soil, no PCR product was amplified. An experimental dilution series of extracted DNA rigorously demonstrated that DNA from an equivalent single specimen of the target virus-vector species, X. diversicaudatum and/or X. index, could be detected amongst 1000 equivalent non-targetX. vuittenezi. Also, extracted DNA from an equivalent single target specimen was detected when added to DNA extracted from the overall soil nematode community. The primers were assessed further by using serial mixtures of actual nematodes rather than extracted DNA to simulate field soil. Using this method, a single target nematode could be detected amongst 200 non-target specimens. Given their specificity, sensitivity and reliability, it appears that these diagnostic primers will be of great benefit to phytosanitary/quarantine services related to the viticulture industry.     

Nicotinamide dehydrogenase subunit 4 analysis of Xiphinema diversicaudatum and Xiphinema simile (Nematoda: Longidoridae)

Regions within the mitochondrial gene encoding for the nicotinamide dehydrogenase subunit 4 (nad4) were characterized to evaluate the extent of genetic variation within and among Xiphinema diversicaudatum and Xiphinema simile populations. Four different sequence variants of nad4 were determined among eight populations of X. diversicaudatum and three variants among three populations of X. simile. Nucleotide variation was detected in 28 of 411 bp (2.43 to 4.87 %) in X. diversicaudatum and in three of 395 bp (0.25 to 0.76 %) in X. simile. This represents the first study based on the characterization of the nad4 gene for the analysis of population genetic of two Xiphinema species.     

Integrative identification and molecular phylogeny of dagger and needle nematodes associated with cultivated olive in Tunisia

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated olive (cvs. Chemlali and Chétoui) in Tunisia were investigated. Morphological and morphometrical studies identified three Longidorus and six Xiphinema species, with frequencies of prevalence as following: Longidorus africanus (23.0 %), L. euonymus (4.5 %), L. glycines (13.7 %), Xiphinema conurum (13.7 %), X. italiae (36.4 %), X. meridianum (13.7 %), X. pachtaicum (18.2 %), X. robbinsi (9.1 %), and Xiphinema sp. (4.5 %). The three Longidorus species were reported for the first time in Tunisia, in addition to two species of Xiphinema (viz. X. meridianum and X. robbinsi). Molecular characterisation using D2-D3 expansion regions of 28S rRNA and ITS1-rRNA was carried out and Bayesian inference analysis was used to reconstruct phylogenetic relationships among these species and with other longidorids. Twenty-five new D2-D3 of 28S rRNA gene sequences were obtained in the present study, seven for Longidorus and 18 for Xiphinema spp., as well as 14 new ITS1 rRNA gene sequences (seven for Longidorus and seven for Xiphinema spp.).     

Influence of environmental factors on glyphosate efficacy when applied to Avena fatua or Urochloa panicoides

Glasshouse experiments were conducted to determine the effects of soil moisture content, irradiance, temperature and relative humidity on the efficacy of glyphosate applied to six isogenic lines of Avena fatua L. (wild oat) and four near-isogenic lines of Urochloa panicoides Beauv. (liverseed grass). The variables examined were four soil moisture conditions (29%, 42%, 55% and 100% of field capacity), two levels of irradiance (400 and 800 μmol m^?2 s^?1), two levels of relative humidity (>92% and 65%) and four temperature regimes (20/15, 25/20, 30/25 and 35/30 °C: day/night), representing the environmental conditions of winter or summer fallows in the north-east grain region of Australia. The efficacy of 360 g acid equivalent ha^?1 glyphosate was greatest under well-watered (100% of field capacity), warm (30/25 °C for A. fatua and 35/30 °C for U. panicoides) and humid (>92/90%) conditions. The efficacy was least under severe water stress (29% of field capacity), warm (30/25 °C for A. fatua and 35/30 °C for U. panicoides) and moderately humid (65/60%) conditions. Efficacy was not altered by the level of irradiance nor was it different between isogenic lines.     

Integrative diagnosis and molecular phylogeny of dagger and needle nematodes of olives and grapevines in the island of Crete,Greece, with description of Xiphinema cretense n. sp. (Nematoda,Longidoridae)

The occurrence and geographic distribution of longidorid nematode species inhabiting the rhizosphere of cultivated and wild olive and grapevine in Crete Island were investigated. Morphological and morphometrical studies identified five Longidorus and six Xiphinema species, with frequencies of prevalence (for wild and cultivated olives and grapevines, respectively) as follows: Longidorus closelongatus (2.0–13.3 %), L. cretensis (1.0–6.7 %), L. moesicus (13.3 % only in grapevines), L. orientalis (3.3 % only in grapevines), L. pseudoelongatus (7.0 % only in olives), Xiphinema cretense n. sp. (3.0 % only in olives), X. index (3.0–23.3 %), X. israeliae (6.3 % only in olives), X. italiae (3.3–10.0 %), X. pachtaicum (26.7–42 %) and X. simile (3.3 % only in grapevines). Xiphinema cretense n. sp. is characterized by a body size 3,872–6,135 μm long, lip region anteriorly rounded, separated from the rest of the body by a depression, odontostyle and odontophore 140.6 and 80.3 μm long respectively, vulva position at 46.0–50.5 %, female tail 31.0–38.0 μm long, nearly hemispherical with curvature essentially dorsal and with a tip completely rounded or presenting a very short bulge, c ratio (119.1–186.9), c’ ratio (0.7–0.8). Molecular characterisation using D2-D3 expansion regions of 28S rRNA, 18S rRNA and ITS1-rRNA was carried out and maximum likelihood and Bayesian inference analysis were used to reconstruct phylogenetic relationships among these species and with other longidorids.     

A high temperature requirement for after ripening of imbibed dormant Poa annua L. seeds

Freshly harvested seeds of Poa annua L. collected in south Louisiana were stored in moist soil at seven temperatures between 5°C and 35°C. At monthly intervals, seed lots were removed and germinated at each of the seven temperatures. Seed were dormant for at least 1 month at all test temperatures. Seeds stored for 2 months at 30 and 35°C showed conditional dormancy; there was 100% germination at 10 or 15°C, and poorer germination at 5 or 20°C. Seeds started to lose viability after 2 months at 35°C and were dead after 7 months. In seeds stored at 10–30°C, there were increased percentages and a wider range of germination temperatures as storage time or storage temperatures increased. Seeds stored at 10°C remained dormant for 9 months, but by 12 months of storage the seeds germinated only at 5 or 10°C. Nearly all seeds stored at the same temperatures in air dry soil remained dormant for 6 months, regardless of storage temperature. These results differ from other reports of low temperatures breaking seed dormancy in Poa annua L. and suggest an adaptation to subtropical climates.     

The degradation of pirimiphas-methyl on stored grains

The degradation of pirimiphos-methyl on grains stored at moisture contents of ≈ 13 and ≈20% has been investigated. Wheat and rice grains were sprayed with [¹⁴C]- pirimiphos-methyl and stored at 20°C under controlled humidity conditions for periods of 12 and 24 weeks. At a moisture content of 13%, degradation was slow, so that at least 70% of the radioactive residue was unchanged pirimiphos-methyl after 24 weeks. Faster degradation occurred at 20% moisture content, but the major component of the radioactive residue (at least 52%) was still unchanged pirimiphos-methyl after 24 weeks. Two major degradation products were formed on the grains; one (18–23.4% of the residue) was characterised as 2-diethylamino-6-methyl-pyrimidin-4-ol, and the other (1.0–9.7% of the residue) was converted to the first by acid hydrolysis. In addition, trace amounts of O-2-ethylamino-6-methyl-pyrimidin-4-yl OO-dimethyl phosphorothioate, 2-ethylamino-6-methyl-pyrimidin-4-ol, 2- amino-6-methyl-pyrimidin-4-ol and an unidentified compound were also detected.     

Viability of herbicide-treated seeds of Eupatorium odoratum L.

The 75–80% viability of Eupatorium odoratum seeds was found to be affected by the temperature of the environment where they were stored. The loss of viability increased with longer periods of storage. Seeds stored under — 10°C or 8°C still retained their viability after 26 months, those stored under 27.5°C or 37°C lost their viability during the same period. There was a gradual loss of viability of herbicide-treated seeds stored under — 10°C or 8°C. The rate of loss was greater for herbicide-treated seeds stored under 27.5°C and 37°C and eventually led to the complete loss of viability after a short storage period of up to 13 months. EPTC and molinate had the greatest effects on seed viability.     

Effect of environmental factors on herbicidal activity of diphenamid

Diphenamid (N,N-dimethyl-2,2-diphenylacetamide) in an aqueous solution in plastic bottles was partially detoxified when exposed to sunlight for 1 week. Varying spray volumes from 300 to 1,800 I/ha did not have an appreciable effect on the phytotoxicity of diphenamid, sprayed on a coarse or fine soil surface. The marked dissipation of diphenamid which occurred from the soil surface was attributed to photodecomposition and volatilization. Diphenamid phytotoxicity was greater when the first irrigation after spraying was applied in four increments of 100 m³/ha or two increments of 200 m¹/ha than when it was applied in a single 400 m¹/h watering; the latter caused more leaching of the herbicide. The diphenamid fraction leached out of a 4-cm soil layer increased as the organic matter content in the soil decreased, from 25% in peat (22.3% o.m.) to >88% in sandy loam (0.9% o.m.). The herbicidal activity remaining after leaching was lower in sandy loam and in peat than in soil with medium organic matter content (11.6% and 6.2%). Diphenamid degradation rate in soil at 50% field capacity moisture level, increased when temperature was increased from 10° to 30°C. After 4 months of incubation at 10°C, 40-50% of the original herbicide was detoxified, while at 20° and 30°C the loss exceeded 90%. Within the range of day-temperatures of 10° to 40°C in soil and of 10° to 35°C in nutrient solution, diphenamid phytotoxicity to tomato seedlings increased with temperature.     

Viruses transmitted by Xiphinema species in the Netherlands

Samenvatting De in Nederland voorkomendeXiphinema-soorten te wetenX. diversicaudatum enX. coxi, werden bestudeerd op hun vermogen om vier virussen over te brengen (Tabel 1).X. coxi bleek een vector van het tobacco ringspot virus te zijn. Het strawberry latent ringspot virus werd voor de eerste maal in Nederland overgebracht metX-diversicaudatum.     

GROWTH,TUBER FORMATION AND SPREAD OF CYPERUS ROTUNDUS L. FROM SINGLE TUBERS*

Summary. Single tubers of Cyperus rotundus L. were planted at intervals over the year. Plant growth was slow and sprouting of tubers was inhibited at temperatures below 20°C, but tubers overwintered at temperatures above freezing point. In the warm season, plant growth and tuber formation rate closely followed air temperature and tubers were forming within 1 month from planting. No inflorescence appeared during the cool season. In autumn-planted C. rotundus grown in containers, the ratio of aerial to subterranean weight decreased from 1·1 in December to 0·2–0·4 in summer. The weight of tubers in mid-summer was about 10 times more than that present in December. Tubers formed at ail times of year and at various locations on plants sprouted readily in laboratory tests (76–100% sprouting). C. rotundus planted in March at wide spacings was grown in field conditions free of other plant competition for 20 months. Within 2 months the plants had spread to 90 cm. At the end of the first and the second summer of growth, the mean area of one plant was 7·6 m² and 56·7 m², respectively, and patches had expanded then by 2·8 m and 5·4 m, respectively, from the initial shoot. After 20 months of growth all tubers were present within the 0–40 cm soil depth, 60–70% of them in the 0–20 cm layer. About 30% of the tubers were within 1 m and 60% within 2 m of the plant centre. Under the patch centre there were about 1000 tubers per m² with 0·3 kg dry weight; in the upper 20 cm more than 3500 tubers weighing 0·9 kg were present per m³ of soil. Croissance, formation de tubercules et propagation de Cyperus rotundus L. issu de tubercules uniques     

Nematode response to methyl bromide and 1,3‐dichloropropene soil fumigation at different temperatures

Several heat‐based methods, such as soil solarization, are being developed as alternative practices for managing soil‐borne pests and pathogens. The effectiveness of these practices is often inconsistent or marginal, thus commanding the need for their integration with other methods. The main objective of this study was to determine synergistic interaction between soil fumigants and temperature. Soil infested with citrus nematode Tylenchulus semipenetrans was exposed to methyl bromide or 1,3‐dichloropropene at various temperatures. Fumigant degradation was concurrently measured and concentration‐time index (ct) was calculated and correlated to the recovered nematode population. In untreated soil, nematode survival was not affected by temperatures of 20–30 °C, but was strongly reduced at ≥ 40 °C. In fumigated soil, nematode suppression was much greater at 30 °C than at 20 °C, and the ct required for nematode elimination at 30 °C was < 50% of that needed at 20 °C for both fumigants. These results suggest that these fumigants became more active with increasing temperature in the sub‐lethal temperature range. It also implies that, when integrated with a heat‐based practice, reduced rates of fumigants may provide adequate pest control, thus minimizing the environmental input of chemical fumigants. © 2000 Society of Chemical Industry     

Dormancy and viability of Phalaris minor seed in a rice–wheat cropping system

Seed placement, soil temperature and soil moisture content influenced the process of after-ripening in Phalaris minor seeds. Seeds of P. minor collected from the soil just after wheat harvesting exhibited higher germination than seeds from P. minor threshed directly. There was a pronounced impact of periodic inhabitation of seed into the soil on germination after its dispersal. Germination was strongly inhibited when the seed was kept in soil at more than field capacity (FC) or in water. Maximum germination of seed incubated in soil at FC occurred at 30°C while a temperature of 40°C favoured after-ripening of seed when mixed with dry soil or kept dry without any medium. Release from conditional dormancy was quicker in the seed retrieved from the soil kept at 20°C than at 10°C. Seed release from conditional dormancy and germination increased with a rise in temperature from 30 to 40°C when the seed was retrieved from incubation in soil at FC for 70 days. The seed kept immersed in water was least responsive to a rise in temperature. Seed recovered from dry soil, or kept without any medium, responded quickly at both temperatures. Light enhanced the germination of Phalaris minor seed. The seedbank subjected to rice (Oryza sativa) field management conditions lost vigour in comparison with the seed stored in laboratory. There was significant variability in seed viability when exposed to differential water management conditions in rice.     

Temperature requirements for after-ripening in buried seeds of four summer annual weeds

Freshly matured, seeds of the four summer annuals Ambrosia artemisiifolia, Polygonum pensylvanicum, Amaranthus hybridus and Chenopodium album were buried in soil at (12/12 h) daily thermoperiods of 15/6, 20/10, 25/15, 30/15 and 35/20°C and at a constant temperature of 5°C. After 0, 1, 3 and 5 months, seeds of each species at each temperature were exhumed and tested at a 14-h daily photoperiod at all six temperatures. Fresh seeds of A. artemisiifolia and P. pensylvanicum did not germinate at any temperature, those of A, hybridus germinated to 4 and 64% at 30/15 and 35/20°C, respectively, and those of C. album to 11–20% at 25/15, 30/15 and 35/20°C. Seeds of A. artemisiifolia and P. pensylvanicum, which germinate only in spring, required exposure to low (5, 15/6°C) temperature to after-ripen completely (i.e., to gain the ability to germinate over a wide range of temperatures), and little or no after-ripening occurred at high (25/15, 30/15 and 35/20°C) temperatures. Seeds of A. hybridus and C. album, which germinate in spring and summer, required exposure to low temperature to after-ripen completely, but at high temperatures they rapidly gained the ability to germinate at high temperatures. Regardless of the burial temperatures and species, when after-ripening occurred, seeds firs germinated at high and then at low temperatures. The minimum germination temperature for a species decreased with after-ripening temperature and with an increase in the length of the burial period.     

Chlorpyrifos degradation in soil at termiticidal application rates

Chlorpyrifos [O,O-diethyl O-(3,5,6-trichloro-2-pyridyl) phosphorothioate] is an organophosphorus insecticide applied to soil to control pests both in agricultural and in urban developments. Typical agricultural soil applications (0.56 to 5.6 kg ha^?1) result in initial soil surface residues of 0.3 to 32 μg g^?1. In contrast, termiticidal soil barrier treatments, a common urban use pattern, often result in initial soil residues of 1000 μg g^?1 or greater. The purpose of the present investigation was to understand better the degradation of chlorpyrifos in soil at termiticidal application rates and factors affecting its behaviour. Therefore, studies with [¹⁴C]chlorpyrifos were conducted under a variety of conditions in the laboratory. Initially, the degradation of chlorpyrifos at 1000 μg g^?1 initial concentration was examined in five different soils from termite-infested regions (Arizona, Florida, Hawaii, Texas) under standard conditions (25°C, field moisture capacity, darkness). Degradation half-lives in these soils ranged from 175 to 1576 days. The major metabolite formed in chlorpyrifos-treated soils was 3,5,6-trichloro-2-pyrid-inol, which represented up to 61% of applied radiocarbon after 13 months of incubation. Minor quantities of [¹⁴C]carbon dioxide (< 5%) and soil-bound residues (? 12%) were also present at that time. Subsequently, a factorial experiment examining chlorpyrifos degradation as affected by initial concentration (10, 100, 1000 μg g^?1), soil moisture (field moisture capacity, 1.5 MPa, air dry), and temperature 15, 25, 35°C) was conducted in the two soils which had displayed the most (Texas) and least (Florida) rapid rates of degradation. Chlorpyrifos degradation was significantly retarded at the 1000 μg g^?1 rate as compared to the 10 μg g^?1 rate. Temperature also had a dramatic effect on degradation rate, which approximately doubled with each 10°C increase in temperature. Results suggest that the extended (3–24 + years) termiticidal efficacy of chlorpyrifos observed in the field may be due both to the high initial concentrations employed (termite LC ₅₀ = 0.2– 2 μg g^?1) and the extended persistence which results from employment of these rates. The study also highlights the importance of investigating the behaviour of a pesticide under the diversity of agricultural and urban use scenarios in which it is employed.     

Persistence of pronamide in soil

Under field conditions, there was little loss of herbicidal activity following spring application of pronamide when the soil temperature remained below about 13°c, but under normal summer conditions loss was rapid (half-life 2–4 weeks). The rate of loss was retarded when the surface soil became very dry. After autumn application, there was no change in activity during the winter months and assays on samples taken in the following spring showed that little leaching had taken place from the surface 5 cm. In laboratory studies, breakdown was shown to follow first-order kinetics. Half-lives at 10% soil moisture were 29 days at 23°c, 63 days at 15°c and 140 days at 8°c. At 23°c the half-life was extended to 52 days when the soil moisture content was reduced by half.