Lima Bean Plants Research Articles

Combined application of humic substances and PGPR inoculated and co-inoculated in plants of Phaseolus lunatus (L.) and Leucaena leucocephala (Lam.) de Wit

The objective of this research was to evaluate the effect of inoculation and co-inoculation of rhizobia and Azospirillum brasilense combined with humic substances (HS) in growth promotion of Phaseolus lunatus (lima bean) and Leucaena leucocephala (leucaena). For this, experiments in a greenhouse with the cultivation of each plant species were carried out. A randomized complete block experimental design with five repetitions was followed. Plant seeds were sown and then inoculated with rhizobia and co-inoculated with A. brasilense. Subsequently, HS were added at the dose recommended by the manufacturer. In the experiments with both plants, control treatments with the addition of nitrogen (N) with or without HS were used. After 45 days, the shoot dry mass (SDM), root dry mass (RDM), shoot-accumulated N (Nac) and relative efficiency index (REI) were determined. In addition, the mass of dry nodules (MDN) in the lima bean plants and the number of nodules (NN) in leucaena plants were determined. The results showed that in the lima bean and leucaena plants there was a greater increase in SDM, RDM and Nac in treatments that received HS and co-inoculation with rhizobia and A. brasilense than in treatments that were inoculated only with rhizobia and HS and in treatments that received N and HS compared to addition of N and isolated inoculation of rhizobia. The combined application of HS and rhizobia in co-inoculation with A. brasilense had a greater effect on the increase of MDN in lima bean and NN in leucaena, than in treatments where only rhizobia with HS were added. These results indicate the existence of potential interaction of the use of HS with the co-inoculation of rhizobia and A. brasilense, showing promise for the production of sustainable agricultural crops.

Rhizobia-legume symbiosis mediates direct and indirect interactions between plants, herbivores and their parasitoids

Microorganisms associated with plant roots significantly impact the quality and quantity of plant defences. However, the bottom-up effects of soil microbes on the aboveground multitrophic interactions remain largely under studied. To address this gap, we investigated the chemically-mediated effects of nitrogen-fixing rhizobia on legume-herbivore-parasitoid multitrophic interactions. To address this, we initially examined the cascading effects of the rhizobia bean association on herbivore caterpillars, their parasitoids, and subsequently investigated how rhizobia influence on plant volatiles and extrafloral nectar. Our goal was to understand how these plant-mediated effects can affect parasitoids. Lima bean plants (Phaseoulus lunatus) inoculated with rhizobia exhibited better growth, and the number of root nodules positively correlated with defensive cyanogenic compounds. Despite increase of these chemical defences, Spodoptera latifascia caterpillars preferred to feed and grew faster on rhizobia-inoculated plants. Moreover, the emission of plant volatiles after leaf damage showed distinct patterns between inoculation treatments, with inoculated plants producing more sesquiterpenes and benzyl nitrile than non-inoculated plants. Despite these differences, Euplectrus platyhypenae parasitoid wasps were similarly attracted to rhizobia- or no rhizobia-treated plants. Yet, the oviposition and offspring development of E. platyhypenae was better on caterpillars fed with rhizobia-inoculated plants. We additionally show that rhizobia-inoculated common bean plants (Phaseolus vulgaris) produced more extrafloral nectar, with higher hydrocarbon concentration, than non-inoculated plants. Consequently, parasitoids performed better when fed with extrafloral nectar from rhizobia-inoculated plants. While the overall effects of bean-rhizobia symbiosis on caterpillars were positive, rhizobia also indirectly benefited parasitoids through the caterpillar host, and directly through the improved production of high quality extrafloral nectar. This study underscores the importance of exploring diverse facets and chemical mechanisms that influence the dynamics between herbivores and predators. This knowledge is crucial for gaining a comprehensive understanding of the ecological implications of rhizobia symbiosis on these interactions.

Two-way plant-mediated interactions between a plant parasitic nematode and a foliar herbivore arthropod

Interactions between belowground and aboveground heterotrophic communities with no direct physical contact can be connected by the plant as a mediator. Plants respond to the attack of herbivores by producing a suite of defensive compounds that can affect the choice and performance of other herbivores. The aim of this study was to determine the impact of one herbivore's activity on the acceptability of that plant to another species of herbivore. Two herbivores were tested; root-knot nematode, Meloidogyne incognita (RKN), a belowground plant-parasitic nematode, and the two-spotted spider mite, Tetranychus urticae (TSSM), an aboveground folivore. We conducted herbivore preference and performance tests on Lima bean (LB) (Phaseolus lunatus cv. Henderson) as an optimal host for TSSM, and tomato (Solanum lycopersicum cv. Rutgers) which is a sub-optimal host for TSSM but optimal host for RKN. We used two-choice glass olfactometers to measure the response of RKN to plants that were exposed to TSSM versus a clean LB plant. RKN infected the clean plants at a significantly greater rate than the TSSM exposed plants. TSSM preference was measured, using leaf discs and two-choice olfactometers containing a RKN infected plant versus a clean plant at different days post-inoculation (DPI) of the RKN. TSSM preferred the clean plants to those with 25-day old RKN infections on LB, but preferred RKN infected tomato plants at 1 DPI. We also tested the effect of the inoculation (1 DPI) of the entomopathogenic nematode (EPN) Steinernema carpocapsae on tomato plants and TSSM preferred the EPN inoculated plants. We carried out a non-choice performance test for TSSM on both LB and tomato on plants inoculated with RKN versus clean plants and observed no effect of RKN exposure on TSSM performance. This research shows that plants can mediate interactions between below and aboveground herbivores that share the same plant.

Rhizobacteria control damping-off and promote growth of lima bean with and without co-inoculation with Rhizobium tropici CIAT899.

Rhizoctonia solani compromises the production of lima bean, an alternative and low-input food source in many tropical regions. Inoculation of bacterial strains has been used, but research on their biocontrol and growth promotion potential on lima bean is scarce. The objective of this study was to evaluate the effects of inoculation with rhizobacterial strains of the genera Bacillus, Brevibacillus, Paenibacillus, Burkholderia, Pseudomonas, and Rhizobium in combination or not with N2-fixing Rhizobium tropici on the control of damping-off disease and growth promotion in lima bean plants. Greenhouse experiments were conducted to evaluate the inoculation with bacterial strains with biocontrol potential in combination or not with R. tropici in substrate infected with R. solani CML 1846. Growth promotion of these strains was also assessed. Strains of Brevibacillus (UFLA 02-286), Pseudomonas (UFLA 02-281 and UFLA 04-885), Rhizobium (UFLA 04-195), and Burkholderia (UFLA 04-227) co-inoculated with the strain CIAT 899 (Rhizobium tropici) were the most effective in controlling R. solani, reducing the disease incidence in 47-60% on lima bean. The promising strains used in the biocontrol assays were also responsive in promoting growth of lima bean under disease and sterile conditions. A positive synergistic effect of co-inoculation of different genera contributed to plant growth, and these outcomes are important first steps to improve lima bean production.

First report of Rhizoctonia solani AG 4 causing brown bean of lima bean in Delaware.

Lima bean production has been an economically valuable staple in Delaware agriculture for almost a century, with annual revenue approaching 8 million dollars (USDA-NASS, 2019; Evans et al. 2007). From 2019-2021, lima beans displaying symptoms of brown discoloration, referred to as "brown bean" were observed in the green baby lima variety 'Cypress' across multiple commercial and research fields. Symptoms were present in approximately 1-5% of beans and not visible until pods were opened for harvest. Thirty-seven symptomatic beans were collected and surface disinfested in 0.85% sodium hypochlorite for 30 s, rinsed in sterile deionized water for 30 s, sectioned into four pieces and plated onto potato dextrose agar (PDA) amended with 50 µg/ml penicillin G and streptomycin sulfate. Petri dishes were incubated at 23ºC and observed for colony morphology. Pure cultures were obtained with tan colonies that had mycelia with right angle branching and septations near the branch, consistent with the description of Rhizoctonia solani Kuhn (Sneh et al. 1991). DNA extraction and pathogen identification was confirmed by sequencing of the internal transcribed spacer (ITS) region of nuclear ribosomal DNA using primers ITS4/ITS5 (White et al. 1990) for thirty-seven isolates collected in 2019 and 2020. Isolates were identified as Rhizoctonia solani AG 4 (99.9% sequence identity with GenBank Accession [MN106359.1].) A representative isolate was selected to complete Koch's postulates and the sequence was deposited in GenBank as accession number MW560551. To observe colonization ability, 10 detached pods were sterilized in 75% EtOH for 60 s, then rinsed in Milli-Q water. The detached pods were divided among two 150 mm Petri dishes containing a single 150 mm filter paper saturated with Milli-Q water. Five 1 mm2 agar plugs colonized with the representative R. solani isolate were placed 0.5 cm apart along the length of the pod. Plates were sealed with parafilm and left at room temperature. Control pods were kept in identical conditions but inoculated using clean agar plugs. The trial was repeated and a second trial was conducted on 12 attached asymptomatic pods from C-Elite Select lima bean plants at the succulent seed stage to complete Koch's Postulates. Pods were surface disinfested with 70% ethanol. Three attached pods were wounded with the tip of a sterile scalpel blade where a colonized agar plug was placed and loosely wrapped with a thin parafilm layer to maintain contact. Three attached pods not wounded were also inoculated with a colonized agar plug and wrapped by parafilm. Three wounded and non-wounded pods received clean agar plug controls. Both attached and detached pods were kept at room temperature for one week until symptoms began to manifest on the pod surfaces, at which point the beans from infected pods were removed and placed on PDA, three to a plate. In the attached assay, all beans of both wounded and non-wounded pods developed symptoms. The plates were stored in identical conditions and monitored for 5 days until tan colonies were observed. Culture morphology was consistent with the original isolate in all beans. Sequencing of the ITS region confirmed identity as R. solani AG-4. No symptoms were observed on control pods or seeds. Rhizoctonia solani is most frequently associated with symptoms of root rot (Sharma-Poudyal et al. 2015), but no stem symptoms are associated with the late season "brown bean" that has been observed throughout production in recent years. To our knowledge, this is the first report of Rhizoctonia solani AG 4 causing symptoms of brown bean of lima bean in Delaware. In preliminary observations, symptoms seem to be worse in pods that could have had contact with the soil directly or via rain splash. This disease cannot be detected until pods are split open, which has potential to reduce lima bean quality at harvest. Further monitoring should be conducted to quantify yield impacts and develop appropriate preventative and curative techniques.

Early damage enhances compensatory responses to herbivory in wild lima bean.

Damage by herbivores can induce various defensive responses. Induced resistance comprises traits that can reduced the damage, while compensatory responses reduce the negative effects of damage on plant fitness. Timing of damage may be essential in determining the patterns of induced defenses. Here, we tested how timing and frequency of leaf damage affect compensatory responses in wild lima bean plants in terms of growth and seed output, as well as their effects on induced resistance to seed beetles. To this end, we applied mechanical damage to plants at different ontogenetical stages, at one time point (juvenile stage only) or two time points (seedling and juvenile stage or juvenile and reproductive stage). We found that plants damaged at the seedling/juvenile stage showed higher compensatory growth, and seed output compared to plants damaged only at the juvenile stage or juvenile/reproductive stage. Seeds from plants damaged at the juvenile and juvenile/reproductive stages had fewer beetles than seeds from undamaged plants, however this was driven by a density dependent effect of seed abundance rather than a direct effect of damage treatments. We did not find differences in parasitism rate by parasitoid wasps on seed beetles among plant treatments. Our results show that damage at the seedling stage triggers compensatory responses which implies that tolerance to herbivory is enhanced or primed by early damage. Herbivory often occurs at several time points throughout plant development and this study illustrates that, for a full understanding of the factors associated with plant induced responses in a dynamic biotic environment, it is important to determine the multitrophic consequences of damage at more than one ontogenetical stage.

Top‐down cascading effects of seed‐feeding beetles and their parasitoids on plants and leaf herbivores

Abstract When feeding on a plant, herbivorous insects alter the quality of the plant as a food source. This affects other organisms interacting with the same plant. These so‐called ‘plant‐mediated interactions’ can be altered by parasitoids that attack the herbivores. So far, this research area has mainly focussed on interactions at the leaf level, and very little is known about plant‐mediated interactions via seeds. It is still poorly understood if seeds that survive insect damage have fewer resources to allocate to plant growth and defence against leaf herbivores, and whether parasitoids that kill seed‐feeding insects mitigate such negative effects. Using seeds of wild lima bean plants Phaseolus lunatus, we studied the effect of the intensity of infestation by seed beetles Zabrotes subfasciatus and their parasitoids Stenocorse bruchivora on the following parameters under lab conditions: seed mass and germination, plant growth and defensive compounds (cyanogenic glycosides and flavonoids) and performance of a leaf herbivore species Spodoptera latifascia. In addition, we performed a field experiment using seeds with or without insect damage to investigate the consequences on plant performance and fitness in the wild. Seed beetle infestation had an overall negative impact on seed germination. Lab experiments revealed that damaged seeds produced plants with slower growth and reduced concentration of defensive compounds, which increased the performance of the leaf herbivores. Effects of seed‐feeding on seed germination and plant growth were attenuated by parasitism, resulting in a net increase of the number of viable offspring. In the field, we observed that seed damage impaired germination, delayed flowering time and increased leaf herbivory. Our results show that plant‐mediated interactions between insect herbivores are not limited to leaf herbivores, but extend to seed herbivores. In our study system, parasitoids had no apparent effect on these interactions, despite their strong beneficial effects on germination and plant performance. These findings confirm the long‐lasting consequences of indirect plant‐mediated interactions in a community‐wide ecological context. Furthermore, they contribute to a better understanding of the important but understudied effects of parasitoids on plant fitness. Read the free Plain Language Summary for this article on the Journal blog.

Viability and receptivity of reproductive structures in Lima bean (Phaseolus lunatus L.) for improvement programs

Artificial hybridization in Phaseolus lunatus L. is still little used in genetic improvement programs in Brazil due to the low establishment percentages. This work aimed to evaluate pollen viability and stigma receptivity in different Lima bean genotypes throughout the day. Accessions ‘Roxinha -MT’ and ‘Cana Longa-PB’ were used in the experiment. Five flower buds were collected from each genotype at 6:00 a.m., 10:00 a.m., 2:00 p.m., and 6:00 p.m. The stigma receptivity was determined using 3% hydrogen peroxide, whereas pollen viability was determined with 2% acetocarmine colouration. The genotype and time factors acted independently on the percentage of viable and non-viable pollen grains and stigma receptivity. The genotype ‘Roxinha’ presented 88.54% viable pollen grains, a value 84.27% higher than that of the ‘Cana Longa’ genotype. This percentage varied as a function of time, with the highest means observed at 6:00 a.m. Stigma receptivity was influenced only by the time of the day, undergoing a great reduction in receptivity, from 88% in the early morning to slightly more than 40% around noon. These results indicate that 6:00 a.m. is the most suitable time to perform artificial crosses in Lima bean plants.

Bacterial type III effector-induced plant C8 volatiles elicit antibacterial immunity in heterospecific neighbouring plants via airborne signalling.

Upon sensing attack by pathogens and insect herbivores, plants release complex mixtures of volatile compounds. Here, we show that the infection of lima bean (Phaseolus lunatus L.) plants with the non‐host bacterial pathogen Pseudomonas syringae pv. tomato led to the production of microbe‐induced plant volatiles (MIPVs). Surprisingly, the bacterial type III secretion system, which injects effector proteins directly into the plant cytosol to subvert host functions, was found to prime both intra‐ and inter‐specific defense responses in neighbouring wild tobacco (Nicotiana benthamiana) plants. Screening of each of 16 effectors using the Pseudomonas fluorescens effector‐to‐host analyser revealed that an effector, HopP1, was responsible for immune activation in receiver tobacco plants. Further study demonstrated that 1‐octen‐3‐ol, 3‐octanone and 3‐octanol are novel MIPVs emitted by the lima bean plant in a HopP1‐dependent manner. Exposure to synthetic 1‐octen‐3‐ol activated immunity in tobacco plants against a virulent pathogen Pseudomonas syringae pv. tabaci. Our results show for the first time that a bacterial type III effector can trigger the emission of C8 plant volatiles that mediate defense priming via plant–plant interactions. These results provide novel insights into the role of airborne chemicals in bacterial pathogen‐induced inter‐specific plant–plant interactions.

Herbivory and jasmonate treatment affect reproductive traits in wild Lima bean, but without transgenerational effects.

PremisePlant responses to herbivores and their elicitors include changes in traits associated with phenology, defense, and reproduction. Induced responses by chewing herbivores are known to be hormonally mediated by the jasmonate pathway and can cascade and affect late‐season seed predators and pollinators. Moreover, herbivore‐induced plant responses can be transmitted to the next generation. Whether herbivore‐induced transgenerational effects also apply to phenological traits is less well understood.MethodsHere, we explored responses of wild lima bean plants (Phaseolus lunatus) to herbivory and jasmonate treatment and possible transgenerational effects of herbivore‐induced early flowering. In a controlled field experiment, we exposed lima bean plants to herbivory by leaf beetles or methyl jasmonate sprays (MJ). We then compared plant development, phenology, reproductive fitness and seed traits among these treatments and undamaged, untreated control plants.ResultsWe found that MJ and leaf herbivory induced similar responses, with treated plants growing less, flowering earlier, and producing fewer seeds than undamaged plants. However, seed size, phenolics and cyanogenic glycosides concentrations did not differ among treatments. Seed germination rates and flowering time of the offspring were similar among maternal treatments.ConclusionsOverall, the results confirm that responses of lima bean to herbivory by leaf beetles are mediated by jasmonate; however, effects on phenological traits are not transmitted to the next generation. We discuss why transgenerational effects of herbivory might be restricted to traits that directly target herbivores.

Avoidance of the Plant Hormone Cis-Jasmone by Aedes aegypti Depends On Mosquito Age in Both Plant and Human Odor Backgrounds

Adults of many mosquito species feed on plants to obtain metabolic energy and to enhance reproduction. Mosquitoes primarily rely on olfaction to locate plants and are known to respond to a range of plant volatiles. We studied the olfactory response of the yellow fever mosquito Aedes aegypti to methyl jasmonate (MeJA) and cis-jasmone (CiJA), volatile compounds originating from the octadecanoid signaling pathway that plays a key role in plant defense against herbivores. Specifically, we investigated how Ae. aegypti of different ages responded to elevated levels of CiJA in two attractive odor contexts, either derived from Lima bean plants or human skin. Aedes aegypti females landed significantly less often on a surface with CiJA and MeJA compared to the solvent control, CiJA exerting a stronger reduction in landing than MeJA. Odor context (plant or human) had no significant main effect on the olfactory responses of Ae. aegypti females to CiJA. Mosquito age significantly affected the olfactory response, older females (7–9 d) responding more strongly to elevated levels of CiJA than young females (1–3 d) in either odor context. Our results show that avoidance of CiJA by Ae. aegypti is independent of odor background, suggesting that jasmonates are inherently aversive cues to these mosquitoes. We propose that avoidance of plants with elevated levels of jasmonates is adaptive to mosquitoes to reduce the risk of encountering predators that is higher on these plants, i.e. by avoiding ‘enemy-dense-space’.

Dispensing a Synthetic Green Leaf Volatile to Two Plant Species in a Common Garden Differentially Alters Physiological Responses and Herbivory

Herbivore-induced plant volatile (HIPV)-mediated eavesdropping by plants is a well-documented, inducible phenomenon that has practical agronomic applications for enhancing plant defense and pest management. However, as with any inducible phenomenon, responding to volatile cues may incur physiological and ecological costs that limit plant productivity. In a common garden experiment, we tested the hypothesis that exposure to a single HIPV would decrease herbivore damage at the cost of reduced plant growth and reproduction. Lima bean (Phaseolus lunatus) and pepper (Capsicum annuum) plants were exposed to a persistent, low dose (~10 ng/h) of the green leaf volatile cis-3-hexenyl acetate (z3HAC), which is a HIPV and damage-associated volatile. z3HAC-treated pepper plants were shorter, had less aboveground and belowground biomass, and produced fewer flowers and fruits relative to controls, while z3HAC-treated lima bean plants were taller and produced more leaves and flowers than did controls. Natural herbivory was reduced in z3HAC-exposed lima bean plants, but not in pepper. Cyanogenic potential, a putative direct defense mechanism in lima bean, was lower in young z3HAC-exposed leaves, suggesting a growth–defense tradeoff from z3HAC exposure alone. Plant species-specific responses to an identical volatile cue have important implications for agronomic costs and benefits of volatile-mediated interplant communication under field conditions.

English

The aim of this research was to evaluate the capacity of symbiotic efficiency of native rhizobia from soils of the state of Rio Grande do Sul (RS), Brazil in Phaseolus lunatus L. plants. Soil and nodule samples from predominant legume plants such as Desmodium species were collected in seven locations of the state of RS. For the isolation of the rhizobia from the soil samples, lima bean “olho de cabra preto” variety and “leucena” (Leucena leucocephala) and plants were used as baits. Subsequently, the symbiotic characterization of the isolates was performed by inoculation on lima bean plants in vitro conditions. The selected bacterial isolates were evaluated for biological nitrogen fixation efficiency in a greenhouse experiment, being determined after: shoot dry mass, root dry mass, nodule dry mass, nitrogen (N) accumulated shoot and calculated the N fixation relative efficiency index. Among the 28 rhizobia isolates tested, 11 induced nodule formation in lima bean. The rhizobia Plu03 and Plu14 stimulated a greater increase of plant dry mass, nodule dry mass and nitrogen accumulation in the shoot, reflecting in a higher relative efficiency index. These results suggest that the isolates Plu03 and Plu14 are more efficient in promoting growth of lima bean, which could be recommended for future agronomic efficiency studies. Key words: Rhizobia, lima bean, biological nitrogen fixation.

A survey of RNA genome viruses in Lima bean crops of Northeastern Brazil

ABSTRACT Lima bean plants showing mosaic and leaf distortion symptoms, similar to those induced by viruses, were observed in several production areas in the states of Ceara and Piaui, Northeastern Brazil. The aim of this study was to identify RNA genome viruses that infect lima bean. Field research was conducted from 2017 to 2018. Fifty-five symptomatic samples were randomly collected from seven properties in five municipalities and tested by RT-PCR and DNA sequencing with specific or universal primers for two viruses and two virus groups. Four virus species were identified: cucumber mosaic virus (CMV), cowpea mild mottle virus (CPMMV), cowpea aphid-borne mosaic virus (CABMV) and cowpea severe mosaic virus (CPSMV), which had infection levels of 21.8, 52.7, 47.2, and 1.8%, respectively, as well as double and triple infections. The CMV isolates belonged to subgroup IA. The CPMMV isolates had high nucleotide identity with CPMMV isolates from Brazil, USA and Mexico. The CABMV isolates showed moderate nucleotide identity with Brazilian isolates. Only one sample was infected with CPSMV. This is the first record of CPMMV and CPSMV naturally infecting lima bean. Approaches to virus control are discussed.

First Report of Didymella americana on Lima Bean (Phaseolus lunatus) in Delaware and Maryland

Delaware and Maryland have a long history of commercial production of lima bean (Phaseolus lunatus) for the processing market. In 2015, we observed lima bean plants that had irregular tan lesions with reddish borders in two fields in Sussex County, DE. Lesions ranged from 0.4 to 2.0 cm in diameter. Approximately 10% of leaves were symptomatic, and severity ranged between 5 and 10%. Symptomatic leaves (n = 6 to 10) were collected in September and October from each field for isolation and evaluation. Similar symptoms were observed on lima bean plants at the University of Maryland Lower Eastern Shore Research and Education Center in 2018, and additional symptomatic leaves (n = 6) were collected. Isolations were conducted from six diseased leaves per field (n = 18) by placing 2- to 4-mm sections of leaf on potato dextrose agar (PDA). All lesions resulted in isolates that appeared similar in morphology. Hyphae from the representative colonies were placed on water agar, and after 24 h, hyphal tips were excised. The hyphal tip procedure was repeated once to obtain pure cultures. Six isolates from Delaware and one from Maryland were stored at –80°C until use. Morphological evaluation of isolates grown on oat agar (OA), malt extract agar (MEA), and PDA revealed conidial morphology and dimensions consistent with previously published reports of Didymella americana (Aveskamp et al. 2010; Gorny et al. 2016). Two isolates, De15-002 and Md18-06, were selected for molecular analysis. DNA was extracted from colonies grown on OA; the internal transcribed spacer (ITS) region was amplified with primers ITS1/ITS4, and the partial actin locus region was amplified with primers act-512/act-783. Basic local alignment searches compared sequences to the ITS (GenBank accession no. FJ426972) and partial actin (FJ426870) sequences of D. americana reference strain CBS 185.85 (Chen et al. 2017; Gorny et al. 2016). The ITS sequences of isolates De15-002 and Md18-06 (MN698269 and MN944417, respectively) shared a 100% similarity with the reference strain. The partial actin sequences of isolates De15-002 (MN820723) and Md18-06 (MN820722) were also identical and differed from the reference sequence by 1 bp (99% similarity). To fulfill Koch’s postulates, four 3-week-old lima bean plants (cv. C-Elite) were sprayed with a conidial suspension (3 × 10⁵ conidia/ml) from 30-day-old cultures grown on MEA or with distilled water (control). Following 28 days of incubation at approximately 20°C, lesions were observed on all plants that were inoculated with isolates De15-001, De15-002, De15-004, De15-005, De15-007, De15-009, and Md18-06, but not on the water-inoculated control plants. Reisolations (n = 4 per isolate) onto water agar and MEA were examined after 7 days to confirm morphology similar to the original isolates. Based on the morphological and molecular characteristics, the pathogen was identified as D. americana. This is the first report of D. americana on lima bean in the Mid-Atlantic region of the United States. Lima bean is the most widely grown vegetable crop in Delaware, and in 2018, the state led production nationally. A new disease on the crop is important to the agricultural economy of Delaware, and the yield loss potential of this pathogen needs assessment in the region. This species has only been reported in 2016 on lima bean, in New York (Gorny et al. 2016).

Parasitoids of leaf herbivores enhance plant fitness and do not alter caterpillar‐induced resistance against seed beetles

Abstract Organisms of the third trophic level can indirectly interact with plants. However, whether parasitoids of herbivores have a positive effect on plant fitness has been controversial. In addition to possible effects on plant fitness, parasitoid‐mitigated herbivory can modify plant physiological responses and thereby alter the plant‐mediated indirect interactions between different herbivore species. These types of indirect multitrophic interactions remain largely unexplored. Thus, to understand the full effect of the third trophic level on plants, it is necessary to consider the context of the community of interacting species, both herbivores and their enemies. Here, we investigated whether parasitoids of leaf‐feeding caterpillars affect plant fitness (seed quantity and quality) and the consequences for seed‐dwelling insects at the second and third trophic levels through plant‐mediated effects. To test this, we exposed lima bean plants (Phaseolus lunatus), under controlled field conditions, to caterpillars (Spodoptera latifascia) that were unparasitized or parasitized by the parasitoid species Cotesia marginiventris. Later in the season, we measured seed production and infestation by seed beetles and their parasitoids. We found that parasitoids significantly reduced the leaf damage inflicted by the caterpillars, such that the plants suffered no loss in seed production. Yet, parasitoids had no effect on the emergence of seed beetles (Zabrotes subfasciatus and Acanthoscelides obtectus), which was equally reduced in plants attacked by unparasitized and by parasitized caterpillars. Seeds from undamaged plants were significantly more attacked by Z. subfasciatus beetles. Parasitism rates of seed beetle larvae were similar for all treatments. Although parasitized caterpillars did not damage the plants enough to reduce seed production (unlike unparasitized caterpillars), the damage they inflicted induced resistance against other herbivores. Taken together, these results show how parasitoids can indirectly enhance plant fitness in the context of the local multitrophic ecological networks. These findings have significant implications for natural and agricultural systems since they reveal that the indirect interaction between plants and parasitoids can be beneficial in communities with multiple herbivore species.

Role of cyanogenic glycosides in the seeds of wild lima bean, Phaseolus lunatus: defense, plant nutrition or both?

Cyanogenic glycosides present in the seeds of wild lima bean plants are associated with seedling defense but do not affect seed germination and seedling growth. Wild lima bean plants contain cyanogenic glycosides (CNGs) that are known to defend the plant against leaf herbivores. However, seed feeders appear to be unaffected despite the high levels of CNGs in the seeds. We investigated a possible role of CNGs in seeds as nitrogen storage compounds that influence plant growth, as well as seedling resistance to herbivores. Using seeds from four different wild lima bean natural populations that are known to vary in CNG levels, we tested two non-mutually exclusive hypotheses: (1) seeds with higher levels of CNGs produce seedlings that are more resistant against generalist herbivores and, (2) seeds with higher levels of CNGs germinate faster and produce plants that exhibit better growth. Levels of CNGs in the seeds were negatively correlated with germination rates and not correlated with seedling growth. However, levels of CNGs increased significantly soon after germination and seeds with the highest CNG levels produced seedlings with higher CNG levels in cotyledons. Moreover, the growth rate of the generalist herbivore Spodoptera littoralis was lower in cotyledons with high-CNG levels. We conclude that CNGs in lima bean seeds do not play a role in seed germination and seedling growth, but are associated with seedling defense. Our results provide insight into the potential dual function of plant secondary metabolites as defense compounds and storage molecules for growth and development.

Identification of pea leaf distortion virus and Ludwigia leaf distortion betasatellite associated with yellow leaf curl disease of lima bean in Nepal

Lima bean (Phaseolus lunatus) plants exhibiting yellow mosaic and stunted growth symptoms, typical of begomovirus infection, were collected in Rampur city Nepal. Symptomatic plants were shown to be infected by a monopartite begomovirus in association with a betasatellite. The two begomovirus isolates associated with the disease showed 98.7% nucleotide sequence identity to previously characterized isolates of the recently identified begomovirus species Pea leaf distortion virus (PLDV) from Nepal. The associated betasatellite isolates showed highest nucleotide sequence identity at 99% to Ludwigia leaf distortion betasatellite (LuLDB) from Nepal. This is the first identification of PLDV and LuLDB infecting the grain legume Phaseolus lunatus.

Silicon as resistance inducer in to control black aphid Aphis craccivora Koch, 1854 in Phaseolus lunatus lima beans

ABSTRACT In Brazil, there are few records of insects associated with the cultivation of lima beans; among them, there is the black aphid Aphis craccivora Koch, 1854. The objective of this study was to evaluate the effects of silicon application on the resistance induction of lima bean plants, Phaseolus lunatus, to the black aphid A. craccivora. The experiment was conducted in the Entomology Laboratory of the Phytosanitary Sector of Centro de Ciências Agrárias, Universidade Federal do Piauí (UFPI), Brazil. The effects of the following treatments on biological aspects of the insect were evaluated: silicon applied to soil; silicon applied to soil + leaf; silicon applied to leaf; and control, without silicon application. The following biological variables were evaluated: generation period, reproductive period, and the fertility and daily average of produced nymphs per female. Plant silicon and lignin content were also evaluated. A 1% solution of silicic acid (2.0 g of product diluted in 200 mL of water) was applied around the plant stem (on soil), 15 days after emergence. Leaf application was performed with a 1-L spray, 5 days after the soil application. The non-preference of A. craccivora on lima beans was also evaluated. The evaluations were performed after 48 and 72 hours of infestation by counting nymphs and adults at each leaf section. Silicon application reduces nymph production, thereby interfering in the biological aspects of A. craccivora. Therefore, it can be used in cowpea pest management programs.

Induction of resistance to Colletotrichum truncatum in lima bean

ABSTRACT: Anthracnose (Colletotrichum truncatum) is one of the major diseases of the lima bean culture, found in production fields, causing decrease in productivity. The objective of this study was to evaluate the effect of abiotic and biotic inducers applied in lima bean plants to reduce anthracnose severity. Lima bean accessions were used and treated with: the abiotic inducers calcium silicate (Agrosilício Plus) and silicate clay (Rocksil), using the 3 g dose.L-1; the biotic inducer citric biomass extract (Ecolife), at the dose of 3 mL.L-1; and distilled water as control. To evaluate the resistance induction, the results of severity and degree of resistance of fava bean at 7, 11, 15, 19 and 23 days after inoculation were considered according to the scale of grades and classes of reactions. The design was a randomized block in a factorial arrangement 4 × 15 (treatments × hits) with four blocks. Resistance inducers Agrosilicon Plus, Ecolife and Rocksil have potential to be used in the management of anthracnose in fava bean. The studied accesses showed degree of resistance, being able to be explored in relation to obtaining anthracnose resistant cultivars in future breeding programs of this crop.