Impact and management of the eriophyoid mite Trisetacus juniperinus on the evergreen cypress Cupressus sempervirens

Abstract 1 In the Mediterranean region, the eriophyoid mite Trisetacus juniperinus causes considerable damage to the evergreen cypress, Cupressus sempervirens L., particularly in nurseries and young stands, disturbing the apical growth of the tree. 2 The impact of mites on survival and apical growth of two commercial clones of cypress (Agrimed and Bolgheri), as well as the results of differently timed pesticide applications to suppress mite population on newly grafted trees, were evaluated. 3 Mites easily infested clonal scions from rootstocks that were previously infested in the nursery, inducing tip deformation and disturbance of the growth. Apical growth was significantly lower in infested than in control trees 2 years following the graft. 4 Deformed apical buds were left early by mites, which dispersed in the crown and may have incurred high mortality. This is interpreted as a defensive reaction of the cypress to the mite attack, which involves costs resulting in reduced apical growth in both clones. However, mites partly overcame tree defences in the Bolgheri clone. 5 Healthy rootstock and graft material should be used to limit damage and maintain plant growth because natural infestations rarely occur. In case of attack, a pesticide should be applied as soon as possible because precocious mite infestation has a log‐lasting effect on tree growth, with considerable economic damage.     

Biological characteristics ofCryptosporiopsis abietina on Hinoki cypress and its antagonistic effect to other microorganisms

In isolation tests on different parts of sound Hinoki cypress (Chamaecyparis obtusa), the coelomyceteCryptosporiopsis abietina was isolated, mostly from immature cone scales and inner bark areas of branches and main stems. In isolation tests using branches or stems of 3-yr-old seedlings to 35-yr-old adult cypresses from various localities,C. abietina was isolated in 28 to 100% of the cypresses tested. It was also confirmed that the fungus rarely infects seeds of the cypress, and can survive in dried seeds for several years, though it is not a seed-borne fungus. No symptoms appeared in inoculation tests with the fungus on 8-yr-old cypress. In inoculation test in a greenhouse, reliable infection without symptoms was obtained on wounded cypress seedlings. These results reveal thatC. abietina is a common endophytic fungus in Hinoki cypress. Confrontation and volatile antagonistic tests in Petri dishes indicated that some isolates ofC. abietina produce antifungal substances in vitro. On these isolates, the reproduction of a mycophagous nematodeBursaphelenchus xylophilus was retarded. The results of inoculation tests suggest that the possibility ofC. abietina as a primary causal agent of the resinous stem canker of Hinoki cypress is low. The role ofC. abietina in the cypress is briefly discussed.     

Induced resistance inSolanum dulcamara triggered by the gall miteAceria cladophthirus (Acari: Eriophyoidea)

After infestation ofSolanum dulcamara leaves by the eriophyoid gall miteAceria cladophthirus (Nalepa), induced resistance became operative against subsequent eriophyoid attacks. The protective effect, manifested by reduction in plant damage and/or mite proliferation, lasted up to 40 days. When the challenger wasA. cladophthirus, the number and size of lesions decreased significantly but mite mortality was not enhanced. When the rust miteThamnacus solani was used as challenger, both russeting symptoms and mite survival rate were strongly reduced. AsA. cladophthirus-induced resistance is relatively permanent, it opens new prospects for eriophyoid control in the field.     

Utilisation of a tropical bay as a nursery area by sharks of the families Carcharhinidae and Sphyrnidae

Synopsis At least eight species of sharks of the families Carcharhinidae and Sphyrnidae use Cleveland Bay in northern Australia as a communal nursery area.Carcharhinus dussumieri, C. fitzroyensis, C. limbatus andC. tilstoni use the bay as a seasonal primary nursery, with juveniles occurring in it for only a few months each year immediately after birth. Alternatively,Carcharhinus sorrah, Rhizoprionodon acutus andR. taylori use the bay as a year-round primary and secondary nursery, with juveniles remaining in it up to the size at maturity. AdultR. taylori also persist in the bay, a behavioural pattern possibly explained by their small maximum size. While present immediately after birth the type of utilisation pattern displayed bySphyrna lewini could not be clarified in this study. Although diets of these species in the bay are similar, there is probably little direct competition for food due to the highly productive habitats in the bay supporting an abundance of food resources. The highest numbers of juveniles occur when prey species are the most abundant, and when temporal separation of some seasonally-occurring species of sharks in effect.     

Spread of plant pathogens and insect vectors at the northern range margin of cypress in Italy

The Mediterranean cypress (Cupressus sempervirens) is a multi-purpose tree widely used in the Mediterranean region. An anthropogenic range expansion of cypress has taken place at the northern margin of the range in Italy in recent decades, driven by ornamental planting in spite of climatic constraints imposed by low winter temperature. The expansion has created new habitats for pathogens and pests, which strongly limit tree survival in the historical (core) part of the range. Based on the enemy release hypothesis, we predicted that damage should be lower in the expansion area. By comparing tree and seed cone damage by pathogens and pests in core and expansion areas of Trentino, a district in the southern Alps, we showed that tree damage was significantly higher in the core area. Seed cones of C. sempervirens are intensively colonized by an aggressive and specific pathogen (the canker fungus Seiridium cardinale, Coelomycetes), associated with seed insect vectors Megastigmus wachtli (Hymenoptera Torymidae) and Orsillus maculatus (Heteroptera Lygaeidae). In contrast, we observed lower tree damage in the expansion area, where a non-aggressive fungus (Pestalotiopsis funerea, Coelomycetes) was more frequently associated with the same insect vectors. Our results indicate that both insect species have a great potential to reach the range margin, representing a continuous threat of the arrival of fungal pathogens to trees planted at extreme sites. Global warming may accelerate this process since both insects and fungi profit from increased temperature. In the future, cypress planted at the range margin may then face similar pest and pathogen threats as in the historical range.     

Resistance and virulence interactions between two white pine species and blister rust in a 30-year field trial

We monitored infection and mortality of full-sib families of sugar pine (SP) and western white pine (WWP) selected for different mechanisms of resistance to white pine blister rust for more than 30 years in a field test in northern California. Natural infection was enhanced by interplanting alternate host Ribes spp. among test seedlings. Parents of the families were from three geographic provinces, representing the northern Rocky Mountains of Idaho (WWP), the Cascade Range of Oregon and Washington (WWP), and the Siskiyou Mountains of California and Oregon (SP). Several families of each pine species had major gene resistance (MGR), or genes for partial resistance (PR), or both types. Under the chronic epidemic conditions that prevailed, susceptible controls of both species became infected and died within a few years. Families with MGR segregated in expected Mendelian ratios (susceptibility/resistance), but each R gene—Cr1 in sugar pine and Cr2 in western white pine—was eventually defeated by biotypes of the rust with specific virulence to it (vcr1 and vcr2, respectively). WWP families from Idaho, which lacked Cr2, performed the best overall, ranging from 44 to 64% rust-free; a few WWP families from the Cascades, both with and without Cr2, performed comparably. Families with PR were not specifically vulnerable to vcr1 or vcr2. PR was less frequent in SP than WWP, and SP families had steeper infection rate curves than WWP. Combining PR in pedigrees with MGR may prolong protection of MGR by inhibiting selection and epidemic increase of vcr genes in the rust.     

Oviposition Sites of the Cypress Seed Bug <Emphasis Type="Italic">Orsillus maculatus</Emphasis> and Response of the Egg Parasitoid <Emphasis Type="Italic">Telenomus</Emphasis> gr. <Emphasis Type="Italic">Floridanus</Emphasis>

Phytophagous insects have many strategies to escape parasitoids, for example by hiding eggs into plant tissues, but oviposition in holes made by another insect is rather scarce. The cypress seed bug Orsillus maculatus Fieber (Heteroptera: Lygaeidae) is strictly dependent on the availability of cones of Cupressus sempervirens L. to oviposit. Females lay eggs either in exit holes cut through the cone scale by emerging adults of the cypress seed chalcid, Megastigmus wachtli Seitner (Hymenoptera: Torymidae), or on the inner side of scales of partly open cones. A recently discovered egg parasitoid belonging to the genus Telenomus has been shown to attack bug eggs in both oviposition sites. In this paper we investigated the parasitoid performance according to oviposition sites. Field samplings were conducted in two evergreen cypress orchards located in the south of France. The distribution and condition of the egg patches were compared between the two locations and oviposition sites. Seed bugs preferred to oviposit in emergence holes of M. wachtli, and parasitoid performance was higher in eggs laid on cone scales. The chalcid emergence holes seemed to ensure bug eggs with enemy-free space. Oviposition site selection could be an adaptive strategy to escape parasitoid attack.     

A Field Experiment to Assess the Transplant Success of Salt Marsh Plants into Tidal Wetlands

Field plots were used to assess the restoration potential of three salt marsh species, Juncus maritimus, Leptocarpus similis and Schoenoplectus pungens, within an established salt marsh near Christchurch, New Zealand. A split-block design was used to asses the effects of soil type, (soil from a proposed marsh construction site or estuarine mud from a natural salt marsh), species and plant source (commercial nursery stocks or natural marsh stock). In December, after 9 months, there was no mortality in L. similis and a significant increase in the biomass compared with J. maritimus. S. pungens failed to regenerate following seasonal die-back. Plant biomass was unaffected by soil type, despite occasional higher salinities recorded in the reclamation soil. Natural stocks had significantly higher biomass than nursery stocks (p < 0.01). These results suggest that both L. similis and J. maritimus are appropriate species for transplanting in the Canterbury region and that the soil from the proposed area is suitable for the restoration and construction of tidal wetlands. The success of the transplants could be enhanced by sourcing hardy stock and management regimes may be necessary to reduce salinity extremes and herbivory by rabbits.     

Root Morphology, Stem Growth and Field Performance of Seedlings of Two Mediterranean Evergreen Oak Species Raised in Different Container Types

Outplanting container-grown oak seedlings with undesirable shoot and root characteristics result in poor establishment and reduced field growth. The objective of this study was to determine the influence of container type on both above-and below-ground nursery growth and field performance of one-year old tap-rooted seedlings Quercus ilex L. and Quercus coccifera L. The experiment was conducted in an open-air nursery and the seedlings were grown in three container types. At the end of the nursery, growth period seedlings’ shoot height, diameter (5 mm above root collar), shoot and root biomass, root surface area, root volume and total root length were assessed. Then the seedlings were planted in the field and their survival and growth were recorded for two growing seasons after outplanting. The results showed a difference between the Quercus species in the effect of container type. Q. ilex seedlings raised in paper-pot had significantly greater height, diameter, shoot and root biomass and root volume than those raised in the other two container types. Similarly, Q. coccifera seedlings raised in paper-pot, had significantly greater above-and below-ground growth than those raised in the other two container types. Both oak species showed relatively low survival in the field; the mortality was mainly observed the first year after outplanting, especially after the summer dry period. However, 2 years after outplanting, the paper-pot seedlings of the two oak species showed better field performance.     

Enhancing planting stock quality of Italian cypress (Cupressus sempervirens L.) by pre-cultivation in mini-plugs

Native species are important in ecological restoration of degraded forest ecosystems provided that their ecological engineering potential is exploited. Their failure to get established is usually due to the use of low-quality planting stock and adverse site conditions. Mini-plug transplants are a relatively new and promising production system in the forest nursery area. Objectives of our study were the evaluation of seedling production of Italian cypress (Cupressus sempervirens) in mini-plug containers, the study of the effect of mini-plug density and substrate on planting stock quality, the comparison of the performance between mini-plug seedlings and the standard planting stock produced by Greek nurseries and the evaluation of physiological and morphological variables as predictive indicators of Italian cypress field performance. Our results showed that cypress seedlings could be produced using the mini-plug technique, resulting in higher quality seedlings than the standard stock type. Pre-cultivation of cypress seedlings under favorable conditions for a period of 5 weeks using peat and high mini-plug densities (1800–3500 mini-plugs m^?2) could be recommended. Grading of seedlings by using both easily measured morphological variables (root length, leaf area, root and shoot dry weight) and physiological tests, such as shoot electrolyte leakage, may improve survival and transplanting success. It is concluded that mini-plugs can serve ecological restoration of degraded forest ecosystems more efficiently than standard planting stock.     

The fungus-specificity of mycorrhization helper bacteria (MHBs) used as an alternative to soil fumigation for ectomycorrhizal inoculation of bare-root Douglas-fir planting stocks with Laccaria laccata

Mycorrhization helper bacteria (MHBs) isolated and selected from the Douglas fir-Laccaria laccata symbiotic system have previously been shown to be fungus-specific: they promote ectomycorrhizal establishment of Laccaria laccata but inhibit mycorrhiza formation by other fungi. In this paper, two experiments in a nursery producing two years-old bare-root Douglas-fir planting stocks confirm the specificity of MHBs under field conditions. They also show that, by selectively helping the introduced L. laccata against the resident symbionts, MHBs are an interesting alternative (safer and easier) to soil fumigation for the success of routine controlled mycorrhization of planting stocks in forest nurseries.     

Variability of haemocyte and haemolymph parameters in European flat oyster Ostrea edulis families obtained from brood stocks of different geographical origins and relation with infection by the protozoan Bonamia ostreae

A research project to compare productive traits (growth and mortality), disease susceptibility and immune capability between Ostrea edulis stocks was performed. This article reports the results on the immune capability and its relation with infection by the intrahaemocytic protozoan Bonamia ostreae. Four to five oyster spat families were produced from each of four European flat oyster populations (one from Ireland, one from Greece and two from Galicia, Spain) in a hatchery. The spat were transferred to a raft in the Ría de Arousa (Galicia) for on growing for 2 years. Total haemocyte count (THC) and differential haemocyte count (DHC) were estimated monthly through the second year of growing-out. Three types of haemocytes were distinguished: granulocytes (GH), large hyalinocytes (LHH) and small hyalinocytes (SHH). Significant correlations between the mean relative abundance of GH and SHH of the families and the mean prevalence of B. ostreae, the overall incidence of pathological conditions and the cumulative mortality of the families were found; these correlations supported the hypothesis that high %GH and low %SHH would enhance oyster immune ability and, consequently, would contribute to lower susceptibility to disease and longer lifespan. Infection by B. ostreae involved a significant increase of circulating haemocytes, which affected more markedly the LHH type. The higher the infection intensity the higher the %LHH. This illustrates the ability of B. ostreae to modulate the immune responses of the O. edulis to favour its own multiplication. A significant reduction of the phenoloxidase activity in the haemolymph of oysters O. edulis infected by B. ostreae was observed. Nineteen enzymatic activities in the haemolymph of O. edulis and Crassostrea gigas (used as a B. ostreae resistant reference) were measured using the kit api ZYM^®, Biomerieux. Qualitative and quantitative differences in enzyme activities in both haemocyte and plasma fractions between B. ostreae noninfected O. edulis from different origins were recorded. However, no clear positive association between enzyme activity and susceptibility to bonamiosis was found. The only enzyme detected in the resistant species C. gigas that was not found in the susceptible one O. edulis was β-glucosidase (in plasma). B. ostreae infected O. edulis showed significant increase of some enzyme activities and the occurrence of enzymes that were not detected in noninfected oysters. These changes could be due to infection-induced enzyme synthesis by the host or to enzyme synthesis by the parasite.     

Survival ofXanthomonas campestris pv.vesictoria in pepper seeds and roots in symptomless and dry leaves in non-host plants and in the soil

Summary A population ofXanthomonas campestris pv.vesicatoria developed as endophytes in the leaves and rhizosphere of apparently symptomless plants grown under mist but not under dry conditions. The pathogen survired for long periods on, and could be isolated from, the surface of infested dried seeds, inoculated sandy loam soil, dried leaves, and the rhizosphere of pepper and of other non-host plants. In addition, small numbers of the pathogen survived for 18 months in a field previously cropped with pepper diseased with bacterial scab. Healthy nursery or mature plants developed symptoms while growing in soil containing infested leaves, either buried or placed on the soil surface.     

The in vitro screening of 118 Trichoderma isolates for antagonism to Rhizoctonia solani and an evaluation of different environmental sites of Trichoderma as sources of aggressive strains

Trichoderma isolates were collected from different sources and screened for their in vitro parasitism of Rhizoctonia solani. They were grouped according to the different sources and each group compared statistically. 74% of the total isolates collected were regarded as antagonistic to R. solani in vitro. Isolates associated with pine bark source were very aggressive. The most aggressive strains were isolated from soil samples collected under the Speedling^® trays of a commercial seedling nursery.     

<Emphasis Type="Italic">Cupressus</Emphasis> callus and cell suspension cultures: Effect of seiridins on their growth and sensitivity

Summary Cupressus macrocarpa and C. arizonica were examined for callus and cell culture production in vitro. Both species produced callus on agar-solidified MSCY medium supplemented with vitamins, antioxidants, 0.14 μM kinetin (KIN), and 10 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Suspension cultures of both species were established in liquid MSCY medium. Seiridin (SE) and iso-seiridin (ISE), two phytotoxic butenolides produced by Seiridium cardinale, S. cupressi, and S. unicorne, the causal agents of many canker diseases of cypress, were tested on callus or cell suspension cultures. In the medium without other plant growth regulators (PGR), SE promoted cell proliferation of cypress better than ISE, for callus initiation, callus maintenance, and cell suspension cultures. The growth rates of cypress callus tissues and suspension cultures of both cypress species on media containing 50–150 μM SE or ISE were measured. At concentrations of 50 μM and higher, growth rates increased exponentially with the SE concentration. A comparison with KIN and 2,4-D indicated that 50 μM SE promoted growth of callus tissues and cell suspension cultures more than 100 μM ISE. SE can also interact with, or counteract, KIN and 2,4-D. It was demonstrated that SE could replace KIN in the medium for C. arizonica. SE could be involved in cell enlargement and proliferation processes. The less susceptible cypress species (C. arizonica) had a high content of terpenoids than that of the more susceptible species (C. macrocarpa). SE could be a useful tool as a phytohormonal-like regulator to manipulate physiological changes at the cellular level and as an elicitor of sensitivity or tolerance of cypress germplasm to the phytotoxin.     

Evaluation of the strains of Acinetobacter and Enterobacter as potential biocontrol agents against Ralstonia wilt of tomato

Bacterial wilt (Ralstonia solanacearum) of tomato, Lycopersicon esculentum, causes a considerable amount of damage to tomato in Southern China. Biological control is one of the more promising approaches to reduce the disease incidence and yield losses caused by this disease. Based on antagonistic activity against R. solanacearum and three soil-borne fungal pathogens as well as biocontrol efficacy in the greenhouse, two bacterial strains Xa6 (Acinetobacter sp.) and Xy3 (Enterobacter sp.) were selected out of fourteen candidates as potential biocontrol agents. In order to find a suitable antagonist inoculation method, we compared the methods of root-dipping with soil-drenching in the aspects including rhizocompetence, biocontrol efficacy, and effect of promoting plant growth under greenhouse conditions. The drenching treatment resulted in a higher biocontrol efficacy and plant-yield increase, and this method was also easier to operate in the field on a large scale. Field trials were conducted for further evaluation of these two antagonistic strains. In both greenhouse and field experiments, the strain Xy3 had a better control effect against bacterial wilt than Xa6 did, while Xa6 caused higher biomass or yield increases. As recorded on the 75th day after treatment in two field experiments, biocontrol efficacy of Xy3 was about 65% in both field trials, and the yield increases caused by Xa6 were 32.4 and 40.7%, respectively, in the two trials. This is the first report of an Acinetobacter sp. strain used as a BCA against Ralstonia wilt of tomato.     

Two new Ig VH gene families in Oncorhynchus mykiss

Genes encoding the immunoglobulin heavy-chain variable region (Ig VH) in rainbow trout (Oncorhynchus mykiss) have been grouped into 11 families. While obtaining a baseline assessment of the various gene families utilized by trout in the production of secreted antibody, we discovered two new families. These proposed Ig VH families, Families XII and XIII, were rarely observed; only two VH sequence types were detected for each new family, suggesting that they may not be commonly used in response to antigens, or that the captive environment may not lead to typical exposures seen in the wild. Additionally, unlike preceding studies, we found at least one representative gene sequence for each of the 11 reported Ig VH gene families, possibly indicating that the repertoire of trout Ig VH gene families may be more universal among different stocks than previously realized. GenBank accession numbers: Family XII—DQ453185 and DQ453150; Family XIII—DQ453153 and DQ453146; others DQ453143, DQ453156, DQ831723, DQ831825.     

A simple model for selection and rapid advancement of transgenic progeny in sorghum

To select agronomically useful transgenic plants, a large number of transgenic events are initially produced, gene transfer confirmed, and advanced to obtain homozygous lines for testing in field trials. Direct in planta assays for identifying the transgene carriers in the segregating progeny are based on the activity of selectable marker gene and are easy, simple and inexpensive. For this purpose, expression of bar gene as measured by tolerance to damage by glufosinate ammonium, the active ingredient in the herbicide BASTA, was investigated. Dose damage curves were generated by leaf paint tests with BASTA on four genotypes of sorghum. Transgenic plants were characterized in terms of sensitivity to the concentration of glufosinate ammonium. In transgenics, symptoms of BASTA swab tests at different growth stages and PCR analysis for cry1B were carried out and correlated. Germination tests could not be employed for large scale evaluation of transgenic progeny because of mortality of tolerant seedlings after transplantation to soil. Based on the above findings, a simple, inexpensive, time-saving, two-step scheme for effective evaluation of transgenics and their progeny containing bar gene as selection marker using BASTA swab tests is described.     

Control of charcoal root rot in <Emphasis Type="Italic">Pinus radiata</Emphasis> nurseries with antagonistic bacteria

The objective of this study was to determine the potential of antagonistic bacteria to control charcoal root rot of coniferous seedlings caused by Macrophomina phaseolina (Tassi) Goid. in forest nurseries. Bacterial isolates were collected from nurseries located between Region Metropolitana and the VIII Region of Chile. Antagonists were initially evaluated in in vitro assays based on the ability to inhibit mycelial growth of M. phaseolina, and subsequently in two trials in a Pinus radiata nursery with a natural infestation of the pathogen. For nursery trials, the isolates were selected according to in vitro and field trial pathogen controls. The bacteria were applied as seed treatments and via water irrigation. The trials were conduced in a completely randomized block design. Among 568 bacterial isolates tested in vitro, 19.8% displayed some capacity to inhibit the mycelial growth of M. phaseolina, with inhibition between 1.7% and 67.6%. In the first nursery trial, Bacillus amyloliquefaciens VII 015, Bacillus pumilus IX 030, Bacillus stearothermophilus TM 008 and other two Bacillus sp. (VI 009 and IX 049) strains, significantly reduced the total, pre- and post-emergency mortality of seedlings, but no isolate reduced the incidence of M. phaseolina in seedlings. In the second trial, Bacillus sp. IX 049, VI 099, B. subtilis (IX 007) and a non-identified isolate V 005, decreased the incidence of charcoal root rot. It is concluded that the best of these bacterial antagonists have the potential to control M. phaseolina in P. radiata nurseries.