Enhanced resistance to soybean cyst nematode Heterodera glycines in transgenic soybean by silencing putative CLE receptors

CLE peptides are small extracellular proteins important in regulating plant meristematic activity through the CLE‐receptor kinase‐WOX signalling module. Stem cell pools in the SAM (shoot apical meristem), RAM (root apical meristem) and vascular cambium are controlled by CLE signalling pathways. Interestingly, plant‐parasitic cyst nematodes secrete CLE‐like effector proteins, which act as ligand mimics of plant CLE peptides and are required for successful parasitism. Recently, we demonstrated that Arabidopsis CLE receptors CLAVATA1 (CLV1), the CLAVATA2 (CLV2)/CORYNE (CRN) heterodimer receptor complex and RECEPTOR‐LIKE PROTEIN KINASE 2 (RPK2), which transmit the CLV3 signal in the SAM, are required for perception of beet cyst nematode Heterodera schachtii CLEs. Reduction in nematode infection was observed in clv1, clv2, crn, rpk2 and combined double and triple mutants. In an effort to develop nematode resistance in an agriculturally important crop, orthologues of Arabidopsis receptors including CLV1, CLV2, CRN and RPK2 were identified from soybean, a host for the soybean cyst nematode Heterodera glycines. For each of the receptors, there are at least two paralogues in the soybean genome. Localization studies showed that most receptors are expressed in the root, but vary in their level of expression and spatial expression patterns. Expression in nematode‐induced feeding cells was also confirmed. In vitro direct binding of the soybean receptors with the HgCLE peptide was analysed. Knock‐down of the receptors in soybean hairy roots showed enhanced resistance to SCN. Our findings suggest that targeted disruption of nematode CLE signalling may be a potential means to engineer nematode resistance in crop plants.     

Alternative products for Pratylenchus brachyurus and Meloidogyne javanica management in soya bean plants

Nematodes stand out among the main soya bean yield limiting factors due to limitations in the main control methods, mainly when they are applied alone. Given the need of finding new control techniques, the aim of this study was to assess the efficiency of products based on biological control agents and on nutrients or products showing potential to induce resistance for Pratylenchus brachyurus and Meloidogyne javanica control in soya bean plants. The experiments were conducted in a greenhouse; nine treatments with six repetitions for each nematode were assessed in two distinct periods. Overall, all treatments have shown potential to control the nematodes in at least one experiment, mainly the ones containing biological control agents (Trichoderma and Bacillus). The vegetative development results were inconclusive; however, some products have shown potential to improve the growth of infected plants.     

Carbendazim resistance and dimethachlone sensitivity of field isolates of Sclerotinia sclerotiorum from oilseed rape in Henan Province,China

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is one of the most important diseases in oilseed rape‐growing areas of China. To determine the frequency of resistance of field isolates of S. sclerotiorum to carbendazim and dimethachlone, a total of 556 isolates from 10 different regions of Henan Province were obtained between 2015 and 2016. The frequency of isolates with a high‐resistance phenotype and a moderate‐resistance phenotype to carbendazim was 69.2% and 10.8%, respectively. However, S. sclerotiorum isolates resistant to dimethachlone were not detected. The baseline sensitivity of S. sclerotiorum to dimethachlone was distributed as a unimodal curve with a mean EC₅₀ value of 0.39 ± 0.09 μg ml^?1 for the inhibition of mycelial growth. Four dimethachlone‐resistant mutants were obtained from 20 wild‐type isolates induced by exposure to increasing concentrations of the fungicide in vitro. The mutants showed high levels of resistance to dimethachlone, with resistance factors that ranged from 179 to 323. Positive cross‐resistance occurred between dimethachlone and procymidone, iprodione, and fludioxonil; however, no cross‐resistance was observed for carbendazim and boscalid. The fitness of the dimethachlone‐resistant mutants was significantly lower than that of the wild‐type isolates, as measured by mycelial growth, hyphal dry weight, sclerotium number and dry weight, and pathogenicity. Additionally, based on osmotic tests, the inhibition of mycelial growth caused by NaCl applied at different concentrations was significantly higher for the dimethachlone‐resistant mutants than for their wild‐type parents.     

Influence of Selected Cultural Practices on Winter Survival of Pratylenchus brachyurus and Subsequent Effects on Soybean Yield

Planting date of soybean, Glycine max, influenced winter survival of Pratylenchus brachyurus in microplots at two locations in North Carolina. Delayed planting resulted in a linear decrease (P = 0.05) in the numbers of P. brachyurus at soybean harvest. Effects of planting date on nematode numbers persisted over winter, indicating that survival in the absence of a host is density independent. Compared with winter fallow, winter wheat, Triticum aestivum, reduced winter survival of P. brachyurus. Subsequent soybean yields were suppressed by the overwintering population of this nematode at one location but not at another.     

Fusarium root rot of coneflower seedlings and integrated control using <Emphasis Type="Italic">Trichoderma</Emphasis> and fungicides

Fusarium root rot (Fusarium spp.) is one of the most important seedling diseases of coneflower (Echinacea spp.) in Alberta greenhouses. Effects of microbial antagonists (Trichoderma spp.) and fungicides, including difenoconazole, fludioxonil, and a mixture of fludioxonil, metalaxyl and difenoconazole, on the management of this disease, were investigated in Alberta. Twenty Trichoderma isolates demonstrated antagonistic activity to Fusarium in agar plate bioassays, with inhibition rates ranging from 44 to 65%. Some Trichoderma isolates significantly ( p < 0.05) reduced disease incidence and severity on seedlings in greenhouse experiments. An in vitro bioassay indicated that difenoconazole and the mixture equally inhibited the growth of both Fusarium and Trichoderma, but, while fludioxonil strongly inhibited the growth of Fusarium, it had little effect on Trichoderma, according to the dose--response models developed ( p < 0.01, R²= 0.902-0.998). Two Trichoderma isolates, T1 and T13 were applied singly or in combination with a low rate of fludioxonil in greenhouse evaluations. The results suggested that fludioxonil and Trichoderma could be integrated into a disease management program for fusarium root rot in coneflower.     

Influence of Planting Date on Population Dynamics and Damage Potential of Pratylenchus brachyurus on Soybean

Planting date was used as a variable to determine the effects of time and different environmental conditions on the population dynamics and damage potential of Pratylenchus brachyurus on soybean at two locations in North Carolina. An initial population slightly less than the damage threshold (275 nematodes/500 cm³ soil) was used to minimize the influence of host damage on this nematode''s population dynamics and to gain greater precision in characterizing factors which influence the damage potential of P. brachyurus to soybean. Equivalent nematode numbers generally resulted in greater yield suppression of soybean in early plantings. Early planting of soybean also resulted in greater (P = 0.01) population densities of P. brachyurus at midseason which often persisted until soybean harvest. Length of time for reproduction and intraspecific competition occurring when soybeans were stunted by the nematode were the most important factors influencing the population dynamics of P. brachyurus.     

A thorough study of a Paratylenchus sp. in glasshouse‐grown lettuce: Characterisation,population dynamics,host plants and damage threshold as keys to its integrated management

In glasshouses practising monoculture of butterhead lettuce in Belgium, high densities of pin nematodes (Paratylenchus spp.) are frequently associated with reduced plant growth. Growers currently apply chemical soil disinfestation measures to manage this problem, although stricter phytosanitary regulations are forcing a shift towards integrated management. Efficient implementation of such management requires knowledge about the factors influencing nematode population dynamics, and the damage threshold for lettuce. The nematode populations in five Belgian glasshouses were monitored for at least 1 year by frequently soil sampling at 0–30 cm and 30–60 cm depth. An undescribed species of Paratylenchus was identified in all glasshouses based on morphological and molecular features. High nematode densities (>20,000 (100 ml soil)^?1) occurred in winter and spring. Chemical soil disinfestation lowered these populations greatly, although up to 14% survived in the deeper soil layer. After soil steaming under negative pressure, no pin nematodes were found. After 2 months of black fallow pin nematode densities were reduced by 50%–76%. Lamb's lettuce, parsley and wild rocket were found to be poor hosts in a pot experiment, while reproduction factors (P_f/P_i) on lettuce cultivars varied between 1 and 3. In three experiments with butterhead lettuce ‘Cosmopolia’ in pots with a series of 9 or 10 densities of Paratylenchus sp. [up to 35,000 (100 ml soil)^?1], no damage to lettuce heads was observed. However, root weight and root quality were reduced, and the corresponding damage thresholds were rather low [1,754 and 362 Paratylenchus sp. (100 ml soil)^?1, respectively]. Management strategies such as crop rotation, soil disinfestation or fallow are recommended to avoid pin nematode population build‐up.     

Assessment of potential antagonists for anthracnose (Colletotrichum gloeosporioides) disease of mango (Mangifera indica L.) in North Western Ethiopia (Pawe)

Mango (Mangifera indica L.) is considered as one of the most popular fruits among millions of people in the tropical area and increasingly in the developed countries. Anthracnose, caused by the fungus Colletotrichum gloeosporioides, is the most important pre- and post-harvest disease of mango. The objective of this research was to evaluate the prevalence of different promising antagonistic Trichoderma and Bacillus spp. on phyloplane of mango in Ethiopia and to evaluate their antagonistic potential against the pathogen. A total of 19 mango fields were surveyed and anthracnose affected all fields. Culture studies on potato dextrose agar for evaluation of antibiosis activity of Trichoderma spp. and Bacillus spp. revealed that they have inhibitory and lytic effect on C. gloeosporioides, which is an indication of their potential biocontrol agent for management of mango anthracnose as an alternative to chemical control. Significant differences (p?<?0.05) were observed among Bacillus isolates in causing lysis of pathogen mycelium, when inoculated on actively growing colony of C. gloeosporioides. Maximum reduction in growth rate of pathogen was observed with Bacillus spp. (B50), which restricted the growth to 2.7?mm compared to 8.3?mm in the control with 67.5% efficacies. There were similar effects (p?<?0.05) among Trichoderma spp. in formation of inhibition zones and lysis by varying degrees up to 59.7% efficacies in reducing linear growth of the pathogen in dual culture.     

Influence of volatile compounds from herbivore‐damaged soybean plants on searching behavior of the egg parasitoid Telenomus podisi

Parasitoids use herbivore‐induced plant volatiles (HIPVs) to locate their hosts. However, there are few studies in soybean showing the mechanisms involved in the attraction of natural enemies to their hosts and prey. The objective of this study was to evaluate the influence of volatile organic compounds (VOCs) of soybean, Glycine max (L.) Merr. (Fabaceae) (cv. Dowling), that were induced after injury caused by Euschistus heros (Fabricius) (Hemiptera: Pentatomidae), on the searching behavior of the egg parasitoid Telenomus podisi Ashmead (Hymenoptera: Scelionidae). Four HIPVs from soybean, (E,E)‐α‐farnesene, methyl salicylate, (Z)‐3‐hexenyl acetate, and (E)‐2‐octen‐1‐ol, were selected, prepared from standards at various concentrations (10^?6 to 10^?1 m ), and tested individually and in combinations using a two‐choice olfactometer (type Y). Telenomus podisi displayed a preference only for (E,E)‐α‐farnesene at 10^?5 m when tested individually and compared to hexane, but they did not respond to the other compounds tested individually at any concentration or when combinations of these compounds were tested. However, the parasitoids stayed longer in the olfactometer arm with the mixture of (E,E)‐α‐farnesene + methyl salicylate at 10^?5 m than in the arm containing hexane. The results suggest that (E,E)‐α‐farnesene and methyl salicylate might help T. podisi to determine the presence of stink bugs on a plant. In addition, bioassays were conducted to compare (E,E)‐α‐farnesene vs. the volatiles emitted by undamaged and E. heros‐damaged plants, to evaluate whether (E,E)‐α‐farnesene was the main cue used by T. podisi or whether other minor compounds from the plants and/or the background might also be used to locate its host. The results suggest that minor volatile compounds from soybean plants or from its surroundings are involved in the host‐searching behavior of T. podisi.     

Sensitivity to boscalid in field isolates of Sclerotinia sclerotiorum from rapeseed in Henan Province,China

Sclerotinia stem rot caused by Sclerotinia sclerotiorum is an important disease of oilseed rape in Henan province of China. Boscalid belongs to succinate dehydrogenase inhibitor (SDHI) fungicides, many of which have strong antifungal activity against S. sclerotiorum. In 2015, a total of 175 isolates of S. sclerotiorum were collected from diseased oilseed rape plants in seven different regions of Henan Province. The EC₅₀ values of 175 isolates of S. sclerotiorum to boscalid ranged from 0.0073 to 0.3880 μg ml^?1, and the mean EC₅₀ value was 0.15 ± 0.09 μg ml^?1. The frequency distribution was unimodal. There was no cross‐resistance between boscalid and carbendazim, procymidone, iprodione, dimethachlone, fludioxonil or fluazinam. Field experiments showed that control efficacies of treatments with boscalid (50% WG) at 225, 300 and 375 g ai ha^?1 were 71%, 81% and 90%, respectively. In contrast, the control efficacy of carbendazim (50% WP) at 1,500 g ai ha^?1 was only 52%.     

Development and evaluation of ForestGrowth‐SRC a process‐based model for short rotation coppice yield and spatial supply reveals poplar uses water more efficiently than willow

Woody biomass produced from short rotation coppice (SRC) poplar (Populus spp.) and willow (Salix spp.) is a bioenergy feedstock that can be grown widely across temperate landscapes and its use is likely to increase in future. Process‐based models are therefore required to predict current and future yield potential that are spatially resolved and can consider new genotypes and climates that will influence future yield. The development of a process‐based model for SRC poplar and willow, ForestGrowth‐SRC, is described and the ability of the model to predict SRC yield and water use efficiency (WUE) was evaluated. ForestGrowth‐SRC was parameterized from a process‐based model, ForestGrowth for high forest. The new model predicted annual above ground yield well for poplar (r² = 0.91, RMSE = 1.46 ODT ha^?1 yr^?1) and willow (r² = 0.85, RMSE = 1.53 ODT ha^?1 yr^?1), when compared with measured data from seven sites in contrasting climatic zones across the United Kingdom. Average modelled yields for poplar and willow were 10.3 and 9.0 ODT ha^?1 yr^?1, respectively, and interestingly, the model predicted a higher WUE for poplar than for willow: 9.5 and 5.5 g kg^?1 respectively. Using regional mapped climate and soil inputs, modelled and measured yields for willow compared well (r² = 0.58, RMSE = 1.27 ODT ha^?1 yr^?1), providing the first UK map of SRC yield, from a process‐based model. We suggest that the model can be used for predicting current and future SRC yields at a regional scale, highlighting important species and genotype choices with respect to water use efficiency and yield potential.     

Studies on Antagonistic Effects of Trichoderma Isolates Against Phytophthora cactorum

Bioassays were used to demonstrate the antibiotic effect of Trichoderma isolates on P. cactorum. When both fungi were grown on benomyl-containing PDA medium, the mycelial growth of Trichoderma was suppressed. However, the production of antibiotics by this fungus remained active, leading to inhibition of the mycelial growth of P. cactorum. The antibiotic effect of Trichoderma on zoospores and cysts was tested on a PDA substrate precultured with Trichoderma on cellophane sheets. On the substrate of some Trichoderma isolates, lysis of zoospores, formation of extracellular vesicles, and hypertrophy of the water expulsion vesicle did occur, both resulting in the death of the zoospores. Conidial suspensions of Trichoderma isolates also induced zoospore lysis. It is presumed that membrane-active peptide antibiotics (peptaibols) are involved in zoospore lysis. The peptaibol paracelsin caused lysins of zoospores at a concentration of 2.5 × 10^?4 M. The effect on cysts depended on the Trichoderma isolate tested and the age of Trichoderma preculture. Old cultures (after beginning of sporulation) affected cysts more severely than young cultures (before sporulation) which usually were not lethal to the cysts but induced preferably microsporangium formation, inhibition of cyst germination, and retardation of germ tube growth.     

Overexpression of a soybean salicylic acid methyltransferase gene confers resistance to soybean cyst nematode

Salicylic acid plays a critical role in activating plant defence responses after pathogen attack. Salicylic acid methyltransferase (SAMT) modulates the level of salicylic acid by converting salicylic acid to methyl salicylate. Here, we report that a SAMT gene from soybean (GmSAMT1) plays a role in soybean defence against soybean cyst nematode (Heterodera glycines Ichinohe, SCN). GmSAMT1 was identified as a candidate SCN defence‐related gene in our previous analysis of soybean defence against SCN using GeneChip microarray experiments. The current study started with the isolation of the full‐length cDNAs of GmSAMT1 from a SCN‐resistant soybean line and from a SCN‐susceptible soybean line. The two cDNAs encode proteins of identical sequences. The GmSAMT1 cDNA was expressed in Escherichia coli. Using in vitro enzyme assays, E. coli‐expressed GmSAMT1 was confirmed to function as salicylic acid methyltransferase. The apparent Km value of GmSAMT1 for salicylic acid was approximately 46 μm . To determine the role of GmSAMT1 in soybean defence against SCN, transgenic hairy roots overexpressing GmSAMT1 were produced and tested for SCN resistance. Overexpression of GmSAMT1 in SCN‐susceptible backgrounds significantly reduced the development of SCN, indicating that overexpression of GmSAMT1 in the transgenic hairy root system could confer resistance to SCN. Overexpression of GmSAMT1 in transgenic hairy roots was also found to affect the expression of selected genes involved in salicylic acid biosynthesis and salicylic acid signal transduction.     

Effects of vanillin on the community structures and abundances of Fusarium and Trichoderma spp. in cucumber seedling rhizosphere

Soil microbial communities are crucial to the functioning of agricultural systems but little information is available on the effects of allelochemicals on soil microorganisms in vivo. Cucumber seedlings grown in soil were treated with different concentrations of vanillin (0.02–0.2?μmol/g soil), a phenolic compound with autotoxic activity. The community structures and abundances of Fusarium and Trichoderma spp. in cucumber rhizosphere were analyzed by polymerase chain reaction (PCR)-denaturing gradient gel electrophoresis and quantitative PCR, respectively. Results showed that vanillin changed the community structures of Fusarium and Trichoderma spp. Vanillin decreased the number of bands of Fusarium spp., but increased the number of bands, Shannon–Wiener and evenness indices of Trichoderma spp. (p?Fusarium and Trichoderma spp. (p?Fusarium and Trichoderma spp., and that these two microbial groups showed different responses to vanillin.     

A comparison of isoprene and monoterpene emission rates from the perennial bioenergy crops short‐rotation coppice willow and Miscanthus and the annual arable crops wheat and oilseed rape

Biogenic volatile organic compounds (BVOC) emissions from bioenergy crops may differ from those of conventional crops. We compared emission rates of isoprene and a number of monoterpenes from the lignocellulosic bioenergy crops short‐rotation coppice (SRC) willow and Miscanthus, with the conventional crops wheat and oilseed rape. BVOC emission rates were measured via dynamic vegetation enclosure and GC‐MS analysis approximately monthly between April 2010 and August 2012 at a location in England and from SRC willow at two locations in Scotland. The largest BVOC emission rates were measured from willow in England and varied between years. Isoprene emission rates varied between μg g^?1 h^?1. Of the monoterpenes detected from willow, α‐pinene emission rates were highest (μg g^?1 h^?1), followed by μg g^?1 h^?1 for δ‐3‐carene, μg g^?1 h^?1 for β‐pinene and μg g^?1 h^?1 for limonene. BVOC emission rates measured in Scotland were much lower. Low emission rates of isoprene and α‐pinene were measured from Miscanthus in 2010 (μg g^?1 h^?1 and μg g^?1 h^?1, respectively) but were not detected in subsequent years. Emission rates from wheat of isoprene were negligible but relatively high for monoterpenes (μg g^?1 h^?1 and μg g^?1 h^?1 for α‐pinene and limonene, respectively). No significant emission rates of BVOCs were measured from oilseed rape. The measured emission rates followed a clear seasonal trend. Crude extrapolations based solely on data gathered here indicate that isoprene emissions from willow could correspond to 0.004–0.03% (UK) and 0.76–5.5% (Europe) of current global isoprene if 50% of all land potentially available for bioenergy crops is planted with willow.     

Below‐ground nematode herbivory of resistant soybean cultivars impairs the performances of an above‐ground caterpillar and its parasitoid

1. Interactions between below‐ and above‐ground organisms have attracted interest in recent years, but less is known about the effect of root‐feeding nematodes on above‐ground trophic interactions between herbivores and their natural enemies. 2. This study examined whether the presence of the soybean cyst root nematode Heterodera glycines influences the performance of the above‐ground leaf‐chewing tobacco cutworm Spodoptera litura on soybeans, Glycine max, and whether this in turn affects the success of its parasitoid, Meteorus pulchricornis. Using three soybean cultivars that varied in the level of constitutive resistance to the tobacco cutworm, the study determined whether feeding by the nematode altered the developmental and reproductive performances of the caterpillar and its parasitoid. 3. Root feeding by the nematode slowed tobacco cutworm larval development time and reduced adult body weight. Root feeding by the nematode also had a negative effect on the caterpillar's parasitoid, by prolonging development time, decreasing adult body size and reducing fecundity. These effects increased in a linear trend and varied in magnitude depending on levels of soybean constitutive resistance. 4. These findings demonstrate that root feeding by the nematode can prime soybean plants with negative impacts on their herbivore and its parasitoid, and that the impact may vary in magnitude depending on levels of soybean constitutive defence. The results emphasise the need to integrate soybean constitutive and root nematode‐induced defences for a better understanding of below‐ and above‐ground organism interactions, and to allow insights to be gained into the improvement of soybean integrated pest management programmes.     

Variation in Aggressiveness Components in the Hemileia vastatrix Population in Brazil

The Pucciniomycete fungus Hemileia vastatrix causes leaf rust on coffee trees. The pathogen is responsible for considerable yield losses in susceptible coffee cultivars if appropriate management strategies are not implemented. Rapid spread and epidemics of rust fungi are usually associated with the emergence of new races of the pathogen that overcome resistance or with the emergence of more aggressive populations of the pathogen. In Brazil, coffee production is dominated by susceptible cultivars of Coffea arabica and Coffea canephora. We assessed aggressiveness in 46 populations of H. vastatrix from Minas Gerais and Espírito Santo, two of the most important coffee‐producing states in Brazil. We observed a significant difference in the incubation period between the populations from Minas Gerais and Espírito Santo when 183 single‐pustule isolates were inoculated onto Catuaí Vermelho IAC 44, a susceptible C. arabica cultivar. Variation in aggressiveness components was observed between and within localities. Isolates with longer incubation periods also tended to have longer latent periods, although there was only a low correlation between these two aggressiveness components (r² = 0.34, P = 2.2 × 10^?16). Low‐sporulating isolates also had significantly longer incubation and latent periods. The H. vastatrix population from Minas Gerais and Espírito Santo is structured by the formation of groups of individuals with differential level of aggressiveness. Our results indicate that the variation in aggressiveness of the Brazilian H. vastatrix population may be associated with the geographic coffee‐producing areas.