Production of Bean yellow mosaic virus resistant subterranean clover (Trifolium subterraneum) plants by transformation with the virus coat protein gene

Transgenic lines of subterranean clover were constructed that contained three different Bean yellow mosaic virus (BYMV) coat protein (CP) gene constructs; full-length CP, the core region of the CP, and full-length CP plus the 3′ untranslated region of the viral genome. Transgenic plants containing the full-length and core CP gene constructs showed high and moderate levels of BYMV resistance. Resistance was measured as a lack or amelioration of viral disease symptoms, which was correlated with a reduction in virus levels and yield loss. A range of different resistance phenotypes was observed. They included reduced infection rates, delay and reduction in local lesion development, and delay and reduction in severity of systemic symptom development. Resistance levels were not correlated with transgene mRNA levels and no transgene-encoded protein was detected in any of the transgenic lines. This is the first example of genetically engineered virus resistance in a clover.     

Effects of cereal borders, admixture with cereals and plant density on the spread of bean yellow mosaic potyvirus into narrow-leafed lupins (Lupinus angustifolius)

In studies of virus control measures, field experiments in 1987–1991 investigated the effects of cereal and fallow borders, admixture with cereals and plant density on spread of bean yellow mosaic potyvirus (BYMV) from pastures dominated by subterranean clover (Trifolium subterraneum) into plots of narrow-leafed lupins (Lupinus angustifolius). Virus spread was mainly monocyclic because BYMV killed infected lupin plants and between systemic movement and death there was only a brief period for BYMV acquisition and transmission to other plants by vector aphids. In plots with cereal borders, the rate and extent of BYMV spread into the lupins was decreased; at final assessment the numbers of infected plants were 43–60% less than in plots with fallow borders. Admixture with cereals also decreased the rate and extent of BYMV spread into lupin plots, numbers of infected plants being decreased by 76–96% at the time of final assessment. When lupins were sown at different seeding rates to generate a range of plant densities and weeds were removed, high densities decreased BYMV infection. The higher incidences of BYMV infection in sparse stands were attributed partly to smaller plant numbers and partly to incoming viruliferous vector aphids being more attracted to plants with bare earth around them, than to a plant canopy. BYMV infection decreased grain yield of samples from infected lupin plants by 94–100%. In plots with 34% infection and sparse stands, grain yield was decreased by about one third. Plotted progress curves for the accumulated numbers of alate aphids of the BYMV vector species Acyrthosiphon kondoi and Myzus persicae resembled those for numbers of BYMV infected plants in 1990, but in 1991 only the curve plotted for M. persicae did so. There was a 2 week delay between the curves for aphid numbers and virus counts which reflected the time taken for obvious systemic necrotic symptoms to develop in lupins.     

Somatic Hybrids Between Potato and S. berthaultii Show Partial Resistance to Soil‐Borne Fungi and Potato Virus Y

Three tetraploid somatic hybrid lines produced by protoplast fusion between a dihaploid potato, Solanum tuberosum, cultivar BF15 and the wild potato species Solanum berthaultii were evaluated here for their response to different soil‐borne pathogens, that is Fusarium solani, Pythium aphanidermatum and Rhizoctonia solani as well as to infection by potato virus Y (PVY). Both hybrid and BF15 plants grown in vitro were inoculated with the tested pathogen strains, that is R. solani, P. aphanidermatum, or F. solani. The growth level and disease severity index of these plants were compared to the susceptible commercial cultivar Spunta. A better growth of inoculated hybrid plants and restricted disease symptoms were observed in comparison with the commercial plants. Under glasshouse conditions and after inoculation with R. solani and P. aphanidermatum, improved resistance of the hybrid plants to these pathogens was confirmed. Indeed, these plants showed no significant damage following inoculation and a better development in R. solani‐infected plants. The susceptibility of the hybrid tubers to R. solani, P. aphanidermatum, and to F. solani infection was also determined. A significant reduction of tissue colonisation was observed in all the hybrid lines compared to the cultivated cultivars. The STBc and STBd hybrids also showed improved resistance to the PVY ordinary strain (PVY^o) under glasshouse conditions.     

Climate change is predicted to alter the current pest status of Globodera pallida and G. rostochiensis in the United Kingdom

The potato cyst nematodes Globodera pallida and G. rostochiensis are economically important plant pathogens causing losses to UK potato harvests estimated at £50 m/ year. Implications of climate change on their future pest status have not been fully considered. Here, we report growth of female G. pallida and G. rostochiensis over the range 15 to 25°C. Females per plant and their fecundity declined progressively with temperatures above 17.5°C for G. pallida, whilst females per plant were optimal between 17.5 and 22.5°C for G. rostochiensis. Relative reproductive success with temperature was confirmed on two potato cultivars infected with either species at 15, 22.5 and 25°C. The reduced reproductive success of G. pallida at 22.5°C relative to 15°C was also recorded for a further seven host cultivars studied. The differences in optimal temperatures for reproductive success may relate to known differences in the altitude of their regions of origin in the Andes. Exposure of G. pallida to a diurnal temperature stress for one week during female growth significantly suppressed subsequent growth for one week at 17.5°C but had no effect on G. rostochiensis. However, after two weeks of recovery, female size was not significantly different from that for the control treatment. Future soil temperatures were simulated for medium‐ and high‐emission scenarios and combined with nematode growth data to project future implications of climate change for the two species. Increased soil temperatures associated with climate change may reduce the pest status of G. pallida but benefit G. rostochiensis especially in the southern United Kingdom. We conclude that plant breeders may be able to exploit the thermal limits of G. pallida by developing potato cultivars able to grow under future warm summer conditions. Existing widely deployed resistance to G. rostochiensis is an important characteristic to retain for new potato cultivars.     

Effects of Temperature and Moisture Variables on Brown Rust Epidemics in Sugarcane

Epidemics of brown rust in sugarcane, caused by Puccinia melanocephala, vary in severity between seasons. Natural epidemics were studied to determine the effects of temperature and moisture variables on epidemic onset, severity and decline. Variables were monitored with disease severity in two cultivars, each grown at a different location in Louisiana. Maximum daily temperature was the variable most correlated with seasonal epidemic development and decline. Disease severity was high during 2009 and low during 2010. This contrast allowed evaluation of the effects of conducive and limiting environmental conditions on severity. Lower severity resulted from a combination of unfavourable temperature and leaf wetness conditions that delayed onset then reduced the rate of disease increase. An accumulation of 23–25 days with leaf wetness periods of at least 7 h after the daily minimum temperature exceeded 17°C preceded the onset of disease on young leaves in both severe and mild epidemics. Severe epidemics in both cultivars declined once maximum ambient daily temperature was 32°C or higher. Low and high limiting temperatures determined the initiation and decline of an epidemic, respectively, under Louisiana climatic conditions. The availability of leaf wetness was then an important determinant of disease severity during the epidemic.     

Host-pathogen-environment interactions predict survival outcomes of adult sockeye salmon (Oncorhynchus nerka) released from fisheries

Incorporating host-pathogen(s)-environment axes into management and conservation planning is critical to preserving species in a warming climate. However, the role pathogens play in host stress resilience remains largely unexplored in wild animal populations. We experimentally characterized how independent and cumulative stressors (fisheries handling, high water temperature) and natural infections affected the health and longevity of released wild adult sockeye salmon (Oncorhynchus nerka) in British Columbia, Canada. Returning adults were collected before and after entering the Fraser River, yielding marine- and river-collected groups, respectively (N = 185). Fish were exposed to a mild (seine) or severe (gill net) fishery treatment at collection, and then held in flow-through freshwater tanks for up to four weeks at historical (14°C) or projected migration temperatures (18°C). Using weekly nonlethal gill biopsies and high-throughput qPCR, we quantified loads of up to 46 pathogens with host stress and immune gene expression. Marine-collected fish had less severe infections than river-collected fish, a short migration distance (100 km, 5–7 days) that produced profound infection differences. At 14°C, river-collected fish survived 1–2 weeks less than marine-collected fish. All fish held at 18°C died within 4 weeks unless they experienced minimal handling. Gene expression correlated with infections in river-collected fish, while marine-collected fish were more stressor-responsive. Cumulative stressors were detrimental regardless of infections or collection location, probably due to extreme physiological disturbance. Because river-derived infections correlated with single stressor responses, river entry probably decreases stressor resilience of adult salmon by altering both physiology and pathogen burdens, which redirect host responses toward disease resistance.     

Effect of Elevated Atmospheric CO2 and Temperature Increases on the Severity of Basil Downy Mildew Caused by Peronospora belbahrii Under Phytotron Conditions

Three experimental trials have been carried out on the basil (Ocimum basilicum)–downy mildew (Peronospora belbahrii) pathosystem, under phytotron conditions, to evaluate the effect of simulated elevated atmospheric CO₂ concentrations and temperatures as well as that of their interaction. Six CO₂ and temperature combinations were tested to establish their effect on disease development. The photosynthetic efficiency (PI) and chlorophyll content index (CCI) of the basil plants were monitored throughout the trials. Average disease incidence was 43.8% under standard conditions (18–22°C and 400–450 ppm of CO₂)_, while average disease severity was 22.1%. In the same temperature regime, a doubled level of CO₂ caused a significant increase in both disease incidence and severity. When temperatures ranged between 18 and 26°C, CO₂ at 800–850 ppm increased disease incidence. At the highest temperatures tested, that is at 26–30°C, which are not favourable for downy mildew development, the increase in CO₂ had no significant effect on disease incidence. A decreasing trend of PI was observed for the PI values of the inoculated plants. This trend was particularly pronounced for high CO₂ levels at the end of the experiment. In the same way as for disease development, lower values were recorded for the inoculated plants at the end of the experiment at 18–22°C for both CO₂ concentrations and at 22–26°C for 850 ppm of CO₂. The non‐inoculated plants showed higher photosynthetic efficiency than the inoculated plants. Similar trends were also observed for the CCI, thus confirming that downy mildew incidence and severity, which in particular caused foliar damage at high CO₂ concentrations, led to a decrease in the physiological performances.     

Identification of a bacterial pathogen associated with Porites white patch syndrome in the Western Indian Ocean

Porites white patch syndrome (PWPS) is a coral disease recently described in the Western Indian Ocean. This study aimed to isolate and identify potential pathogens associated with PWPS utilizing both culture and nonculture screening techniques and inoculation trials. A total of 14 bacterial strains (those dominant in disease lesions, absent or rare in healthy tissues and considered potential pathogens in a previous study) were cultured and used to experimentally inoculate otherwise healthy individuals in an attempt to fulfil Henle–Koch's postulates. However, only one (P180R), identified as closely related (99–100% sequence identity based on 1.4 kb 16S RNA sequence) to Vibrio tubiashii, elicited signs of disease in tank experiments. Following experimental infection (which resulted in a 90% infection rate), the pathogen was also successfully re‐isolated from the diseased tissues and re‐inoculated in healthy corals colonies, therefore fulfilling the final stages of Henle–Koch's postulates. Finally, we report that PWPS appears to be a temperature‐dependent disease, with significantly higher tissue loss (anova : d.f. = 2, F = 39.77, P < 0.01) occurring at 30 °C [1.45 ± 0.85 cm² per day (mean ± SE)] compared to ambient temperatures of 28 and 26 °C (0.73 ± 0.80 cm² per day (mean ± SE) and 0.51 ± 0.50 cm² per day (mean ± SE), respectively).     

Relationship Between Ralstonia solanacearum Diversity and Severity of Bacterial Wilt Disease in Tomato Fields in China

A field survey was conducted to determine the relationship between Ralstonia solanacearum diversity and severity of bacterial wilt disease in tomato plants grown in plastic greenhouses. Both vegetative and reproductive stages of the plants were surveyed, and the symptoms were empirically categorized into five scales: 0 (asymptomatic): 1st, 2nd, 3rd and 4th. The bacterial wilt pathogen was isolated from infected plants at each disease scale; pathogenic characteristics and population densities of the bacterial strains were assessed. Two hundred and eighty‐two isolates were identified as R. solanacearum, which were divided into three pathogenic types, virulent, avirulent and interim, using the attenuation index (AI) method and a plant inoculation bioassay. Ralstonia solanacearum was detected in all asymptomatic and symptomatic tomato plants, with population numbers, ranging from 10.5 to 86.7 × 105 cfu/g. However, asymptomatic plants harboured only avirulent or interim R. solanacearum, whereas tomato plants displaying 1st or 2nd disease degree contained interim and virulent strains. Additionally, 3rd and 4th degree plants harboured only virulent strains. The disease was more severe in vegetative‐stage plants (disease severity index (DSI) 0.20) with higher total numbers of interim and virulent R. solanacearum strains than those in reproductive‐stage plants (DSI 0.12). Three pathotypes of R. solanacearum coexisted in a competitive growth system in the tomato field, and their distribution closely correlated with the severity of tomato bacterial wilt.     

Changing fitness of a necrotrophic plant pathogen under increasing temperature

Warmer temperatures associated with climate change are expected to have a direct impact on plant pathogens, challenging crops and altering plant disease profiles in the future. In this study, we have investigated the effect of increasing temperature on the pathogenic fitness of Fusarium pseudograminearum, an important necrotrophic plant pathogen associated with crown rot disease of wheat in Australia. Eleven wheat lines with different levels of crown rot resistance were artificially inoculated with F. pseudograminearum and maintained at four diurnal temperatures 15/15°C, 20/15°C, 25/15°C and 28/15°C in a controlled glasshouse. To quantify the success of F. pseudograminearum three fitness measures, these being disease severity, pathogen biomass in stem base and flag leaf node, and deoxynivalenol (DON) in stem base and flag leaf node of mature plants were used. F. pseudograminearum showed superior overall fitness at 15/15°C, and this was reduced with increasing temperature. Pathogen fitness was significantly influenced by the level of crown rot resistance of wheat lines, but the influence of line declined with increasing temperature. Lines that exhibited superior crown rot resistance in the field were generally associated with reduced overall pathogen fitness. However, the relative performance of the wheat lines was dependent on the measure of pathogen fitness, and lines that were associated with one reduced measure of pathogen fitness did not always reduce another. There was a strong correlation between DON in stem base tissue and disease severity, but length of browning was not a good predictor of Fusarium biomass in the stem base. We report that a combination of host resistance and rising temperature will reduce pathogen fitness under increasing temperature, but further studies combining the effect of rising CO₂ are essential for more realistic assessments.     

Biological properties of necrotic and non-necrotic strains of bean yellow mosaic virus in cool season grain legumes

A collection of 51 bean yellow mosaic virus (BYMV) isolates was transmitted from infected Trifolium subterraneum (subterranean clover) to Lupinus angustifolius (narrow‐leafed lupin) by Myzus persicae (green peach aphid). Depending on isolate and L. angiistifolius genotype used, two distinct responses developed in L. angustifolius plants, either systemic necrosis and plant death or non‐necrotic reactions of varying severity. Ten isolates caused necrosis and plant death in cv. Danja. However, when nine of these were inoculated to breeding line 90L423‐07‐13, seven induced non‐necrotic reactions, while two caused necrosis and plant death. Thirty seven isolates always produced non‐necrotic reactions regardless of genotype of L. angustifolius inoculated. Non‐necrotic and necrotic isolates originally came both from lupins and other species, and the non‐necrotic isolates were no less efficiently transmitted by M. persicae than the necrotic ones. When one isolate of each type was inoculated together to T. subterraneum and nine months later this culture was used as an acquisition source for aphid transmission to L. angustifolius, only the necrotic type was detected. Previous infection of L. angustifolius plants with a non necrolic isolate prevented subsequent infection by a necrotic one. All necrotic and non‐necrotic isolates reacted with BYMV antiserum in ELISA but only two cross‐reacted with antiserum to clover yellow vein virus (CYVV). When selected necrotic and non‐necrotic isolates were inoculated to differential hosts, all behaved like BYMV and not CYVV. When three isolates of each type were transmitted to 11 other cool season grain legume species, except in Cicer arietinum (chickpea), there were no necrotic reactions, but symptom severity varied with the isolate and species inoculated. The two isolates that caused necrosis in C. arietinum did not do so in L. angustifolius. The six isolates from Vicia faba (faba bean) all caused non‐necrotic reactions in L. angustifolius cv. Danja and 90L423‐07‐13. These and two necrotic isolates readily infected five genotypes of V. faba always causing severe symptoms. However, three non‐necrotic isolates from L. angustifolius and a further necrotic isolate were poorly infectious on V. faba in which they generally induced mild symptoms. These results show that at least three strain groups of BYMV can be distinguished by their reactions in different L. angustifolius genotypes, one causing necrosis and death in cv. Danja and 90L423‐07‐13, one causing necrosis and death in Danja but not 90L423‐07‐13, and one causing non‐nccrotic reactions in both. These strain groups could not be distinguished when representative necrotic and non‐necrotic isolates were inoculated to other grain legume species. However, inoculation to V. faba distinguished two other BYMV strain groupings differing in severity of symptoms and ability to infect this species.     

Determining the effectiveness of grazing and trampling by livestock in transmitting white clover mosaic and subterranean clover mottle viruses

Glasshouse and mini-sward experiments were done to determine the relative roles of grazing and trampling by livestock in transmitting white clover mosaic (WC1MV) and subterranean clover mottle (SCMoV) viruses between clover plants in pastures. Wounding due to grazing was simulated by repeatedly cutting plants with serrated scissors (glasshouse) or mowing (mini-swards), while wounding due to trampling was simulated by repeatedly bashing plants with the flat end of a wooden hammer handle (glasshouse) or rolling (mini-swards). In glasshouse experiments, cutting was more effective than bashing in transmitting WC1MV to white clover (Trifolium repens) plants but cutting and bashing transmitted it to subterranean clover (T. subterraneum) plants at similar rates. In an experiment with white clover mini-swards, mowing was more effective than rolling in transmitting WC1MV, and when both were combined, initially spread exceeded that obtained when the spread from mowing and rolling alone was added together. In glasshouse experiments, bashing was more effective than cutting in transmitting SCMoV to subterranean clover plants. In one experiment, neither mowing nor rolling spread SCMoV in mini-swards of subterranean clover. When transmission to subterranean clover cultivars which were ‘susceptible’ or ‘moderately susceptible’ to SCMoV was compared in glasshouse experiments, repeated bashing spread the virus more slowly to the ‘moderately susceptible’ cultivars. When mixed with ruminant saliva, infective sap containing WC1MV or SCMoV was still infective to clover plants after 4 wk storage at room temperature. When infective sap was allowed to dry naturally on a metal surface, SCMoV still infected clover plants when the dried sap was taken up in tap water after 4 but not 14 days, while WC1MV was infective after 24 h but not 4 days. These results suggest that grazing and mowing are more effective than trampling at transmitting WC1MV to white clover plants in pastures, while trampling is more effective at spreading SCMoV to subterranean clover. However, both transmitted WC1MV to subterranean clover at similar rates. Possible reasons for these differences are discussed in relation to differences in clover plant morphology and virus-specific factors.     

Effect of strain-specific hypersensitive resistance on spatial patterns of virus spread

Spatial patterns of spread were compared between strains of Bean yellow mosaic virus (BYMV) that differ in causing systemic necrotic (hypersensitive) or non‐necrotic symptoms in narrow‐leafed lupin (Lupinus angustifolius). Both types of BYMV were spread naturally by aphids from adjacent infected pasture into a large lupin block (‘natural spread site’), or from clover plants introduced as virus sources into two field experiments with lupin. Cumulative spatial data for plants with disease symptoms from a range of times in the growing period were assessed using Spatial Analysis by Distance IndicEs (SADIE). At the‘natural spread site’, with non‐necrotic BYMV, the extent of clustering of plants with symptoms increased gradually over time, while with necrotic BYMV there was less clustering and no increase over time. In both experiments, for the type of BYMV that was introduced into a plot, there was a gradual increase in clustering, but with this being greater with non‐necrotic BYMV. In the second experiment, there was also significant clustering of plants with symptoms of non‐necrotic BYMV in plots without introduced non‐necrotic foci but not for necrotic BYMV in plots without introduced necrotic foci. When clustering data for plants with newly recorded symptoms was tested for spatial association between successive assessment dates, association was positive for both BYMV types though stronger for the non‐necrotic type, declining as the temporal lag increased. Generally, association was strongest for assessments 2–3 wk apart, corresponding approximately to the period for BYMV to move systemically in plants and for obvious symptoms to appear in shoot tips. Contour maps for local association between dates showed that the strongest spatial associations were from coincidence of infection gaps rather than infection patches. The combination of information from clustering and association analysis showed that spread of non‐necrotic BYMV is less diffuse, with considerably more localised infection surrounding the infection sources. This work demonstrates how spatial virus spread can be diminished when hypersensitive (necrotic) resistance is deployed, and the limitations associated with employing hypersensitivity that is strain specific.     

Identification and Pathogenicity of Lasiodiplodia Species from Eucalyptus urophylla × grandis,Polyscias balfouriana and Bougainvillea spectabilis in Southern China

Species of Lasiodiplodia are important pathogens of a wide variety of plants covering a wide geographical distribution. These fungi can be associated with different symptoms such as stem cankers, shoot blights, fruit rots, dieback and gummosis. Diseases caused by Lasiodiplodia were surveyed on Eucalyptus urophylla × grandis, Polyscias balfouriana and Bougainvillea spectabilis in a nursery in southern China. Based on morphology characteristics and phylogenetic analyses of ITS rDNA sequences and translation elongation factor 1‐alpha (TEF‐1α) gene regions, four species of Lasiodiplodia were identified. Lasiodiplodia theobromae was identified from E. urophylla × grandis, P. balfouriana and B. spectabilis. L. hormozganensis, L. iraniensis and L. pseudotheobromae were identified from B. spectabilis. To our knowledge, with the exception of L. theobromae on E. urophylla × grandis, this study represents the first report of these fungi on the host plants. Pathogenicity tests showed that all Lasiodiplodia spp. obtained in this study are virulent to E. urophylla × grandis and B. spectabilis, and L. theobromae was virulent to P. balfouriana.     

Tolerance and overcompensation to infection by Phytophthora infestans in the wild perennial climber Solanum dulcamara

Studies of infection by Phytophthora infestans—the causal agent of potato late blight—in wild species can provide novel insights into plant defense responses, and indicate how wild plants might be influenced by recurrent epidemics in agricultural fields. In the present study, our aim was to investigate if different clones of Solanum dulcamara (a relative of potato) collected in the wild differ in resistance and tolerance to infection by a common European isolate of P. infestans. We performed infection experiments with six S. dulcamara genotypes (clones) both in the laboratory and in the field and measured the degree of infection and plant performance traits. In the laboratory, the six evaluated genotypes varied from resistant to susceptible, as measured by degree of infection 20 days post infection. Two of the four genotypes susceptible to infection showed a quadratic (concave downward) relationship between the degree of infection and shoot length, with maximum shoot length at intermediate values of infection. This result suggests overcompensation, that is, an increase in growth in infected individuals. The number of leaves decreased with increasing degree of infection, but at different rates in the four susceptible genotypes, indicating genetic variation for tolerance. In the field, the inoculated genotypes did not show any disease symptoms, but plant biomass at the end of the growing season was higher for inoculated plants than for controls, in‐line with the overcompensation detected in the laboratory. We conclude that in S. dulcamara there are indications of genetic variation for both resistance and tolerance to P. infestans infection. Moreover, some genotypes displayed overcompensation. Learning about plant tolerance and overcompensation to infection by pathogens can help broaden our understanding of plant defense in natural populations and help develop more sustainable plant protection strategies for economically important crop diseases.     

Effect of temperature on immature development and longevity of two introduced opiine parasitoids on Bactrocera invadens

Temperature‐dependent development, parasitism and longevity of the braconid parasitoids, Fopius arisanus Sonan and Diachasmimorpha longicaudata Ashmed on Bactorcera invadens Drew Tsuruta & White, was evaluated across five constant temperatures (15, 20, 25, 30 and 35°C). Developmental rate decreased linearly with increasing temperature for both the parasitoid species. Linear and Brière‐2 nonlinear models were used to determine the lower temperature threshold at which the developmental rate (1/D) approached zero. For F. arisanus, lower thresholds to complete development estimated with the linear and nonlinear models were 10.1 and 6.9°C, respectively. The total degree‐days (DD) required to complete the development estimated by the linear model for F. arisanus was 360. In D. longicaudata, the linear and nonlinear models estimated lower thresholds of 10.4 and 7.3°C, respectively, and the total DD estimated was 282. In F. arisanus, percentage parasitism differed significantly across all temperatures tested and was highest at 25°C (71.1 ± 2.5) and lowest at 15°C (46.4 ± 1.4). Parasitoid progeny sex ratio was female biased at all temperatures except at 20°C. In D. longicaudata, percentage parasitism was highest at 20°C (52.2 ± 4.0) and lowest at 15°C (27.7 ± 2.5). Parasitoid progeny sex ratio was female biased and similar for all temperatures. Adult longevity of both parasitoids was shortest at 35°C and longest at 15°C, and females lived significantly longer than males at all temperatures tested. Our findings provide some guidance for future mass rearing and field releases of the two parasitoids for the management of B. invadens in Africa.     

Rapid thermal adaptation in photosymbionts of reef‐building corals

Climate warming is occurring at a rate not experienced by life on Earth for 10 s of millions of years, and it is unknown whether the coral‐dinoflagellate (Symbiodinium spp.) symbiosis can evolve fast enough to ensure coral reef persistence. Coral thermal tolerance is partly dependent on the Symbiodinium hosted. Therefore, directed laboratory evolution in Symbiodinium has been proposed as a strategy to enhance coral holobiont thermal tolerance. Using a reciprocal transplant design, we show that the upper temperature tolerance and temperature tolerance range of Symbiodinium C1 increased after ~80 asexual generations (2.5 years) of laboratory thermal selection. Relative to wild‐type cells, selected cells showed superior photophysiological performance and growth rate at 31°C in vitro, and performed no worse at 27°C; they also had lower levels of extracellular reactive oxygen species (exROS). In contrast, wild‐type cells were unable to photosynthesise or grow at 31°C and produced up to 17 times more exROS. In symbiosis, the increased thermal tolerance acquired ex hospite was less apparent. In recruits of two of three species tested, those harbouring selected cells showed no difference in growth between the 27 and 31°C treatments, and a trend of positive growth at both temperatures. Recruits that were inoculated with wild‐type cells, however, showed a significant difference in growth rates between the 27 and 31°C treatments, with a negative growth trend at 31°C. There were no significant differences in the rate and severity of bleaching in coral recruits harbouring wild‐type or selected cells. Our findings highlight the need for additional Symbiodinium genotypes to be tested with this assisted evolution approach. Deciphering the genetic basis of enhanced thermal tolerance in Symbiodinium and the cause behind its limited transference to the coral holobiont in this genotype of Symbiodinium C1 are important next steps for developing methods that aim to increase coral bleaching tolerance.     

Evaluation of Tomato Hybrids Carrying Ty‐1 and Ty‐2 Loci to Japanese Monopartite Begomovirus Species

Tobacco leaf curl Japan virus, Honeysuckle yellow vein mosaic virus and Tomato yellow leaf curl virus are three begomoviruses that infect tomato crops in Japan. Tomato infection by begomoviruses has increased in Japan after the development of a high level of resistance to certain insecticides in some populations of the vector B. tabaci biotypes ‘B and Q’. Ty‐1 and Ty‐2 homozygous tomato hybrids were evaluated for reaction to monopartite begomovirus species in Japan by Agrobacterium‐mediated inoculation. Test plants were evaluated by a disease assessment scale (DAS), varying from 1 = no symptoms to 4 = severe symptoms, and systemic infection was evaluated by polymerase chain reaction (PCR), using specific begomovirus primers for each virus. Ty‐1 hybrids showed tolerance to HYVMV and with a large number of plants being neither virus‐free nor symptom‐free. The response of Ty‐1 hybrids was also resistant to moderately resistant against TbLCJV. The response of Ty‐2 hybrids was resistant to highly resistant against the three monopartite begomoviruses, when compared with susceptible plants.     

Identification and antifungal activity analysis of two biocontrol antagonists to Colletotrichum musae

Postharvest anthracnose of banana caused by Colletotrichum musae is one of the major diseases resulting in huge economic losses worldwide. To control this disease using biocontrol agents, two antagonistic strains SD7 and NB20 with significant inhibitory effects on mycelial growth and conidial germination of C. musae were identified and evaluated in this study. The inhibitory effects of cell‐free culture filtrates of SD7 and NB20 on conidial germination of C. musae were both 100%, and those on mycelial growth of C. musae were 97.7 ± 0.9% and 95.0 ± 0.6%, respectively. The antifungal activities of cell‐free culture filtrates of both strains were still stable after they were stored at 4°C for 6 months. The control efficacies of cell‐free culture filtrates of SD7 and NB20 on postharvest anthracnose of banana were 55.9 ± 4.1% and 33.2 ± 3.9%, respectively. The disease severity (mean scale value) in banana fruit fingers was significantly lower after the treatment with a cultural suspension of the bacterial strain SD7 (1.4 ± 0.49) or actinomycete strain NB20 (2.0 ± 0.63), compared to that in the control (4.8 ± 0.40). After subculturing for 10 generations, the antifungal efficiency of NB20 remained stable, whereas that of strain SD7 declined obviously. Lastly, based on the morphological, physio‐biochemical and molecular characteristics, the bacterial strain SD7 was identified as Burkholderia cepacia, while the actinomycete strain NB20 was identified as Streptomyces katrae. The results from this study will provide the basis for developing an effective and novel biofungicide to control banana anthracnose disease.