Plant-associated fungi affect above- and belowground pest responses to soybean plants.

AIM: The objective of this research was to screen fungal isolates originally isolated from cotton plants and measure their effects on the interactions between soybean and two aboveground pests (cabbage looper; Trichoplusia ni and soybean looper; Chrysodeixis includens) as well as a belowground pest (soybean cyst nematode; Heterodera glycines). METHODS AND RESULTS: For aboveground pests, we measured the leaf area consumed and larval weight. For our belowground pest tests, we measured shoot height, shoot fresh weight, root fresh weight and number of cysts. Out of the 50 fungal isolates tested, we tested 30 fungi in the interaction with cabbage looper, 36 for soybean looper, 41 for soybean cyst nematode. We tested 23 isolates against all pests and identified multiple isolates that significantly changed the response of pests on inoculated soybean plants versus controls. CONCLUSIONS: We identified three fungal isolates that significantly reduced both leaf area consumed aboveground by caterpillars and number of cysts produced belowground by nematodes. These isolates were an Epicoccum italicum, a Chaetomium undulatum and a Stemphylium majusculum. SIGNIFICANCE AND IMPACT OF STUDY: Overall, this study provides important insights into plant-fungal interactions and their effect on both above- and belowground pests. This study also highlights an important first step towards harnessing the potential of microbial inoculates as a tool for integrated pest management in soybeans.

Lessons from the Far End: Caterpillar FRASS-Induced Defenses in Maize, Rice, Cabbage, and Tomato.

Plant defenses to insect herbivores have been studied in response to several insect behaviors on plants such as feeding, crawling, and oviposition. However, we have only scratched the surface about how insect feces induce plant defenses. In this study, we measured frass-induced plant defenses in maize, rice, cabbage, and tomato by chewing herbivores such as European corn borer (ECB), fall armyworm (FAW), cabbage looper (CL), and tomato fruit worm (TFW). We observed that caterpillar frass induced plant defenses are specific to each host-herbivore system, and they may induce herbivore or pathogen defense responses in the host plant depending on the composition of the frass deposited on the plant, the plant organ where it is deposited, and the species of insect. This study adds another layer of complexity in plant-insect interactions where analysis of frass-induced defenses has been neglected even in host-herbivore systems where naturally frass accumulates in enclosed feeding sites over extended periods of time.

Fungal seed treatments of cotton affect boll weevil development.

BACKGROUND: Nonpathogenic fungi associated with plants can enhance plant defenses against stress factors, including herbivory. This study assessed whether cotton plants grown from seeds treated with different fungi affected boll weevil, Anthonomus grandis grandis Boheman, development and reproduction along with plant tolerance. We used whole plants grown from seeds treated with different fungi (Chaetomium globosum TAMU520 and TAMU559, Phialemonium inflatum TAMU490, and Beauveria bassiana) versus non-treated controls to test insect growth, reproduction, and plant tolerance assays in a greenhouse. RESULTS: Regarding boll weevil reproduction, fewer larvae hatched and fewer adults emerged from fungal-treated plants. In addition, the developmental time from oviposition to adult emergence was delayed in the plants treated with all fungi. For plant tolerance, B. bassiana-treated plants attacked by boll weevils shed fewer squares than nonfungal-treated plants. CONCLUSION: Fungal treatments can affect boll weevil performance and reproduction on cotton plants, with potentially negative effects on population growth. Collectively, these results support the potential for cottonseed treatments with fungi as a novel tool for boll weevil management in the field. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

RNA-Seq and molecular docking reveal multi-level pesticide resistance in the bed bug.

BACKGROUND: Bed bugs (Cimex lectularius) are hematophagous nocturnal parasites of humans that have attained high impact status due to their worldwide resurgence. The sudden and rampant resurgence of C. lectularius has been attributed to numerous factors including frequent international travel, narrower pest management practices, and insecticide resistance. RESULTS: We performed a next-generation RNA sequencing (RNA-Seq) experiment to find differentially expressed genes between pesticide-resistant (PR) and pesticide-susceptible (PS) strains of C. lectularius. A reference transcriptome database of 51,492 expressed sequence tags (ESTs) was created by combining the databases derived from de novo assembled mRNA-Seq tags (30,404 ESTs) and our previous 454 pyrosequenced database (21,088 ESTs). The two-way GLMseq analysis revealed ~15,000 highly significant differentially expressed ESTs between the PR and PS strains. Among the top 5,000 differentially expressed ESTs, 109 putative defense genes (cuticular proteins, cytochrome P450s, antioxidant genes, ABC transporters, glutathione S-transferases, carboxylesterases and acetyl cholinesterase) involved in penetration resistance and metabolic resistance were identified. Tissue and development-specific expression of P450 CYP3 clan members showed high mRNA levels in the cuticle, Malpighian tubules, and midgut; and in early instar nymphs, respectively. Lastly, molecular modeling and docking of a candidate cytochrome P450 (CYP397A1V2) revealed the flexibility of the deduced protein to metabolize a broad range of insecticide substrates including DDT, deltamethrin, permethrin, and imidacloprid. CONCLUSIONS: We developed significant molecular resources for C. lectularius putatively involved in metabolic resistance as well as those participating in other modes of insecticide resistance. RNA-Seq profiles of PR strains combined with tissue-specific profiles and molecular docking revealed multi-level insecticide resistance in C. lectularius. Future research that is targeted towards RNA interference (RNAi) on the identified metabolic targets such as cytochrome P450s and cuticular proteins could lay the foundation for a better understanding of the genetic basis of insecticide resistance in C. lectularius.

A Beneficial Plant-Associated Fungus Shifts the Balance toward Plant Growth over Resistance, Increasing Cucumber Tolerance to Root Herbivory.

Plants allocate their limited resources toward different physiological processes, dynamically adjusting their resource allocation in response to environmental changes. How beneficial plant-associated microbes influence this allocation is a topic that continues to interest plant biologists. In this study, we examined the effect of a beneficial fungus, Phialemonium inflatum, on investment in growth and anti-herbivore resistance traits in cucumber plants (Cucumis sativus). We inoculated cucumber seeds with P. inflatum spores and measured several growth parameters, including germination rate, above and belowground biomass, and number of flowers. We also examined plant resistance to adult and larval striped cucumber beetles (Acalymma vitattum), and quantified levels of defense hormones in leaves and roots. Our results indicate that P. inflatum strongly enhances cucumber plant growth and reproductive potential. Although fungus treatment did not improve plant resistance to cucumber beetles, inoculated plants were more tolerant to root herbivory, experiencing less biomass reduction. Together, these findings document how a beneficial plant-associated fungus shifts plant investment in growth over herbivore resistance, highlighting the importance of microbes in mediating plant-herbivore interactions. These findings also have important implications for agricultural systems, where beneficial microbes are often introduced or managed to promote plant growth or enhance resistance.

Proteomic analysis of labial saliva of the generalist cabbage looper (Trichoplusia ni) and its role in interactions with host plants.

Insect saliva is one of the first secretions to come in contact with plants during feeding. The composition and role of caterpillar saliva has not been as thoroughly studied as that of sucking insects. This study focuses on characterizing the proteome of the cabbage looper (Trichoplusia ni) saliva using iTRAQ labeling and LC-MS/MS. We also measured how the saliva proteome changed when larvae were reared on different diets - cabbage, tomato, and an artificial pinto bean diet. We identified 254 proteins in the saliva out of which 63 were differentially expressed. A large percentage (56%) of the proteins identified function in protein metabolism, followed by proteins involved in vesicle transport (6%) and oxidoreductase activity (5%), among other categories. Several proteins identified are antioxidants or reactive oxygen species (ROS) scavengers. Among these ROS scavengers, we identified a catalase and further analyzed its gene expression and enzymatic activity. We also applied commercial, purified catalase on tomato and measured the activity of defensive proteins - trypsin proteinase inhibitor, polyphenol oxidase and peroxidase. Catalase gene expression was significantly higher in the salivary glands of larvae fed on tomato. Also, catalase suppressed the induction of tomato trypsin proteinase inhibitor levels, but not the induction of polyphenol oxidase or peroxidase. These results add to our understanding of proteomic plasticity in saliva and its role in herbivore offense against plant defenses.

Genomics of Lepidoptera saliva reveals function in herbivory.

Lepidoptera herbivores deposit copious amounts of saliva when feeding. Their saliva is produced by the paired mandibular and labial glands and evidence indicates that it may play an important role in allowing an herbivore to establish on its host plant. Genomic studies of Lepidoptera saliva are beginning to reveal the role of saliva in herbivory. Molecules involved in digestion, detoxification, immunity, defense against plant secondary chemicals, chemoreception and so on have been identified using high throughput genomic tools. These genomic tools have also revealed changes that occur in Lepidoptera saliva when caterpillars feed on different host plants. However, there are other factors either biotic or abiotic (e.g., larval stage, larval health, temperature, water stress, etc.) that might also affect its composition. Though further functional and ecological studies are still necessary to fully understand the role of Lepidoptera saliva on herbivory, here we review current trends.

Host plant driven transcriptome plasticity in the salivary glands of the cabbage looper (Trichoplusia ni).

Generalist herbivores feed on a wide array of plants and need to adapt to varying host qualities and defenses. One of the first insect derived secretions to come in contact with the plant is the saliva. Insect saliva is potentially involved in both the pre-digestion of the host plant as well as induction/suppression of plant defenses, yet how the salivary glands respond to changes in host plant at the transcriptional level is largely unknown. The objective of this study was to determine how the labial salivary gland transcriptome varies according to the host plant on which the insect is feeding. In order to determine this, cabbage looper (Trichoplusia ni) larvae were reared on cabbage, tomato, and pinto bean artificial diet. Labial glands were dissected from fifth instar larvae and used to extract RNA for RNASeq analysis. Assembly of the resulting sequencing reads resulted in a transcriptome library for T. ni salivary glands consisting of 14,037 expressed genes. Feeding on different host plant diets resulted in substantial remodeling of the gland transcriptomes, with 4,501 transcripts significantly differentially expressed across the three treatment groups. Gene expression profiles were most similar between cabbage and artificial diet, which corresponded to the two diets on which larvae perform best. Expression of several transcripts involved in detoxification processes were differentially expressed, and transcripts involved in the spliceosome pathway were significantly downregulated in tomato-reared larvae. Overall, this study demonstrates that the transcriptomes of the salivary glands of the cabbage looper are strongly responsive to diet. It also provides a foundation for future functional studies that can help us understand the role of saliva of chewing insects in plant-herbivore interactions.

Cues from chewing insects - the intersection of DAMPs, HAMPs, MAMPs and effectors.

Chewing herbivores cause massive damage when crushing plant tissues with their mandibles, thus releasing a vast array of cues that may be perceived by the plant to mobilize defenses. Besides releasing damage cues in wounded tissues, herbivores deposit abundant cues from their saliva, regurgitant and feces that trigger herbivore specific responses in plants. Herbivores can manipulate the perception mechanisms and defense signals to suppress plant defenses by secreting effectors and/or by exploiting their associated oral microbes. Recent studies indicate that both the composition of herbivore cues and the plant's ability to recognize them are highly dependent upon the specific plant-herbivore system. There is a growing amount of work on identifying herbivore elicitors and effectors, but the most significant bottleneck in the discipline is the identification and characterization of plant receptors that perceive these herbivore-specific cues.

Allelopathic effects of glucosinolate breakdown products in Hanza [Boscia senegalensis (Pers.) Lam.] processing waste water.

Boscia senegalensis is a drought resistant shrub whose seeds are used in West Africa as food. However, the seeds, or hanza, taste bitter which can be cured by soaking them in water for 4-7 days. The waste water resulting from the processing takes up the bitter taste, which makes it unsuitable for consumption. When used for irrigation, allelopathic effects were observed. Glucosinolates and their breakdown products are the potential causes for both the bitter taste and the allelopathic effects. The objectives of this study are to identify and quantify the glucosinolates present in processed and unprocessed hanza as well as different organs of B. senegalensis, to analyze the chemical composition of the processing water, and to pinpoint the causal agent for the allelopathic properties of the waste water. Hanza (seeds without testa), leaves, branches, unripe, and ripe fruits were collected in three populations and subjected to glucosinolate analyses. Methylglucosinolates (MeGSL) were identified in all plant parts and populations, with the highest concentrations being found in the hanza. The levels of MeGSLs in the hanza reduced significantly during the soaking process. Waste water was collected for 6 days and contained large amounts of macro- and micronutrients, MeGSL as well as methylisothiocyanate (MeITC), resulting from the conversion of glucosinolates. Waste water from days 1-3 (High) and 4-6 (Low) was pooled and used to water seeds from 11 different crops to weeds. The High treatment significantly delayed or reduced germination of all the plant species tested. Using similar levels of MeITC as detected in the waste water, we found that germination of a subset of the plant species was inhibited equally to the waste water treatments. This confirmed that the levels of methylisiothiocyanate in the waste water were sufficient to cause the allelopathic effect. This leads to the possibility of using hanza waste water in weed control programs.