Host-Parasitoid Phenology, Distribution, and Biological Control under Climate Change.

Climate change raises a serious threat to global entomofauna-the foundation of many ecosystems-by threatening species preservation and the ecosystem services they provide. Already, changes in climate-warming-are causing (i) sharp phenological mismatches among host-parasitoid systems by reducing the window of host susceptibility, leading to early emergence of either the host or its associated parasitoid and affecting mismatched species' fitness and abundance; (ii) shifting arthropods' expansion range towards higher altitudes, and therefore migratory pest infestations are more likely; and (iii) reducing biological control effectiveness by natural enemies, leading to potential pest outbreaks. Here, we provided an overview of the warming consequences on biodiversity and functionality of agroecosystems, highlighting the vital role that phenology plays in ecology. Also, we discussed how phenological mismatches would affect biological control efficacy, since an accurate description of stage differentiation (metamorphosis) of a pest and its associated natural enemy is crucial in order to know the exact time of the host susceptibility/suitability or stage when the parasitoids are able to optimize their parasitization or performance. Campaigns regarding landscape structure/heterogeneity, reduction of pesticides, and modelling approaches are urgently needed in order to safeguard populations of natural enemies in a future warmer world.

Endophytic insect pathogenic fungi-host plant-herbivore mutualism: elucidating the mechanisms involved in the tripartite interactions.

Various techniques used by crop plants to evade insect pests and pathogen attacks have been documented. Among these, plant defense strategies induced by endophytic insect pathogenic fungi are arguably one of the most discussed. Endophytic fungi frequently colonize plants and inhabit their internal tissues for a portion of their lifespan without producing visible symptoms of the disease. This phenomenon is widespread and diverse in both natural and agricultural ecosystems, and is present in almost all plant organs. Many fungi can obtain nutrients by infecting and killing insects, and this ability has been developed numerous times in different fungal lineages. These species mainly consist of those in the order Hypocreales (Ascomycota), where the generalist insect pathogens, Beauveria sp. (Cordycipitaceae) and Metarhizium sp. (Clavicipitaceae) are two of the most studied endophytic entomopathogenic fungal genera. However, most fungi that kill insects do not survive in the tissues of living plants. The data published thus far show a high degree of variability and do not provide consistent explanations for the underlying mechanisms that may be responsible for these effects. This implies that available knowledge regarding the colonization of plant tissues by endophytic insect pathogenic fungi, the effects of colonization on plant metabolism, and how this contributes to a decrease in herbivore and pathogens damage is limited. To adequately utilize fungal-based products as biological control agents, these products must be effective and the reduction of pests and infection must be consistent and similar to that of chemical insecticides after application. This article discusses this possibility and highlights the benefits and the specific techniques utilized by endophytically challenged plants in invading insect pests and disease pathogens.

Baseline Analysis of Endophytic Fungal Associates of Solenopsis invicta Buren from Mounds across Five Counties of Guangdong Province, China.

Red imported fire ants mounds have been suggested as a potential reservoir for beneficial entomopathogenic fungal species that are vital for more complex roles in the ecosystem aside from infecting the insects. In the current study, the assemblage of fungal symbionts of the red imported fire ants (RIFA) were obtained across five cities in Guangdong Province, China. The sampling areas were selected because of high occurrence of fire ants mounds in the regions. Mound soils, plant debris within mounds, and ants were collected from three sampling locations in each city for potential isolation of entomopathogenic fungal associates of RIFA. All samples were collected during the spring of 2021. Following successful isolation from substrates, the patterns of fungal species composition, and richness were evaluated. In total, 843 isolates were recovered, and based on their phenotypic distinctiveness and molecular characterization based on DNA sequences of multiple loci including the ITS, SSU, and LSU regions, 46 fungal taxa were obtained, including 12 that were unidentified. Species richness and abundance was highest in the mound soils, while the lowest value was recorded from the ant body. As per the different locations, the highest abundance level was recorded in Zhuhai, where 15 fungal taxa were cultivated. The most common taxa across all substrates and locations was Talaromyces diversus. A baseline analysis of the fungal community composition of RIFA would better our understanding on the interactions between these social ants and their associated microbial organisms, and this knowledge in turn would be important for the successful management of the RIFA.

Interspecific competition predicts the potential impact of little fire ant Wasmannia auropunctata (Roger) (Hymenoptera: Formicidae) invasion on resident ants in southern China.

The little fire ant Wasmannia auropunctata (Roger) is a major invasive species that seriously threatens the biodiversity of invaded areas. W. auropunctata was first reported in Chinese mainland in 2022 and its impact on native species is still unknown. To evaluate the impact of W. auropunctata invasion on the ant communities in southern China, a series of interspecific competition experiments were conducted in this study. The individual aggression index and group aggression experiments showed the advantage of W. auropunctata in competition with 5 resident ants under equal worker numbers. When encountering Anoplolepis gracilipes, Camponotus nicobarensis, Tetramorium bicarinatum, Polyrhachis dives, and Solenopsis invicta, W. auropunctata gradually gained a competitive advantage with an increase in its number of workers. In the group aggression experiments with equal worker numbers, there was a negative correlation between the body length and mortality rate of resident ants. The results of the foraging behavior experiments showed that W. auropunctata was able to dominate food resources under competition with Carebara diversa, which also displayed weak competition in the group aggression bioassay. In addition, the abilities to recruit workers and retrieve food were inhibited under competition with S. invicta and T. bicarinatum. The results of the nesting behavior experiments showed that in the 24-h bout of space resource competition, W. auropunctata was dominant over C. diversa, S. invicta, and T. bicarinatum. The results of this study show that W. auropunctata has certain advantages in competition for food and space resources over resident ants in southern China, and some resident ant species may be replaced in the future.

Comprehensive genome-wide identification and expression profiling of ADF gene family in Citrus sinensis, induced by endophytic colonization of Beauveria bassiana.

Endophytic entomopathogenic species are known to systematically colonize host plants and form symbiotic associations that benefit the plants they live with. The actin-depolymerizing factors (ADFs) are a group of gene family that regulate growth, development, and defense-related functions in plants. Systematic studies of ADF family at the genome-wide level and their expression in response to endophytic colonization are essential to understand its functions but are currently lacking in this field. 14ADF genes were identified and characterized in the Citrus sinensis genome. The ADF genes of C. sinensis were classified into five groups according to the phylogenetic analysis of plant ADFs. Additionally, the cis-acting analysis revealed that these genes play essential role in plant growth/development, phytohormone, and biotic and abiotic responses; and the expression analysis showed that the symbiotic interactions generate a significant expression regulation level of ADF genes in leaves, stems and roots, compared to controls; thus enhancing seedlings' growth. Additionally, the 3D structures of the ADF domain were highly conserved during evolution. These results will be helpful for further functional validation of ADFs candidate genes and provide important insights into the vegetative growth, development and stress tolerance of C. sinensis in responses to endophytic colonization by B. bassiana.

Transcriptome analysis reveals differential effects of beta-cypermethrin and fipronil insecticides on detoxification mechanisms in Solenopsis invicta.

Insecticide resistance poses many challenges in insect pest control, particularly in the control of destructive pests such as red imported fire ants (Solenopsis invicta). In recent years, beta-cypermethrin and fipronil have been extensively used to manage invasive ants, but their effects on resistance development in S. invicta are still unknown. To investigate resistance development, S. invicta was collected from populations in five different cities in Guangdong, China. The results showed 105.71- and 2.98-fold higher resistance against fipronil and beta-cypermethrin, respectively, in the Guangzhou population. The enzymatic activities of acetylcholinesterase, carboxylases, and glutathione S-transferases significantly increased with increasing beta-cypermethrin and fipronil concentrations. Transcriptomic analysis revealed 117 differentially expressed genes (DEGs) in the BC-ck vs. BC-30 treatments (39 upregulated and 78 downregulated), 109 DEGs in F-ck vs. F-30 (33 upregulated and 76 downregulated), and 499 DEGs in BC-30 vs. F-30 (312 upregulated and 187 downregulated). Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis revealed that DEGs associated with insecticide resistance were significantly enriched in metabolic pathways, the AMPK signaling pathway, the insulin signaling pathway, carbon metabolism, peroxisomes, fatty acid metabolism, drug metabolism enzymes and the metabolism of xenobiotics by cytochrome P450. Furthermore, we found that DEGs important for insecticide detoxification pathways were differentially regulated under both insecticide treatments in S. invicta. Comprehensive transcriptomic data confirmed that detoxification enzymes play a significant role in insecticide detoxification and resistance development in S. invicta in Guangdong Province. Numerous identified insecticide-related genes, GO terms, and KEGG pathways indicated the resistance of S. invicta workers to both insecticides. Importantly, this transcriptome profile variability serves as a starting point for future research on insecticide risk evaluation and the molecular mechanism of insecticide detoxification in invasive red imported fire ants.

Genome-Wide Identification and Characterization of Toll-like Receptors (TLRs) in Diaphorina citri and Their Expression Patterns Induced by the Endophyte Beauveria bassiana.

Toll-like receptors (TLRs) are pathogen recognition receptors (PRRs), which play key roles in helping the host immune system fight pathogen invasions. Systematic information on TLRs at the genome-wide level and expression profiling in response to endophytic colonization is very important to understand their functions but is currently lacking in this field. Here, a total of two TLR genes were identified and characterized in Diaphorina citri. The TLR genes of D. citri were clustered into five families according to the phylogenetic analysis of different species' TLRs. The domain organization analyses suggested that the TLRs were constituted of three important parts: a leucine-rich repeat (LRR) domain, a transmembrane region (TR) and a Toll/interleukin-1 receptor (TIR) domain. The mRNA expression levels of the two TLR genes (DcTOLL and DcTLR7) were highly regulated in both nymphs and adults of D. citri. These results elucidated the potentiated TLR gene expression in response to endophytically colonized plants. Furthermore, the 3D structures of the TIR domain were highly conserved during evolution. Collectively, these findings elucidate the crucial roles of TLRs in the immune response of D. citri to entomopathogens systematically established as endophytes, and provide fundamental knowledge for further understanding of the innate immunity of D. citri.

Role of Insect Gut Microbiota in Pesticide Degradation: A Review.

Insect pests cause significant agricultural and economic losses to crops worldwide due to their destructive activities. Pesticides are designed to be poisonous and are intentionally released into the environment to combat the menace caused by these noxious pests. To survive, these insects can resist toxic substances introduced by humans in the form of pesticides. According to recent findings, microbes that live in insect as symbionts have recently been found to protect their hosts against toxins. Symbioses that have been formed are between the pests and various microbes, a defensive mechanism against pathogens and pesticides. Insects' guts provide unique conditions for microbial colonization, and resident bacteria can deliver numerous benefits to their hosts. Insects vary significantly in their reliance on gut microbes for basic functions. Insect digestive tracts are very different in shape and chemical properties, which have a big impact on the structure and composition of the microbial community. Insect gut microbiota has been found to contribute to feeding, parasite and pathogen protection, immune response modulation, and pesticide breakdown. The current review will examine the roles of gut microbiota in pesticide detoxification and the mechanisms behind the development of resistance in insects to various pesticides. To better understand the detoxifying microbiota in agriculturally significant pest insects, we provided comprehensive information regarding the role of gut microbiota in the detoxification of pesticides.

Comprehensive Detoxification Mechanism Assessment of Red Imported Fire Ant (Solenopsis invicta) against Indoxacarb.

The red imported fire ant (Solenopsis invicta) is one of the deadliest invasive ant species that threatens the world by disrupting biodiversity, important functions within a natural ecosystem, and community structure. They are responsible for huge economic losses in the infested countries every year. Synthetic insecticides, especially indoxacarb, have been broadly used to control S. invicta for many years. However, the biochemical response of S. invicta to indoxacarb remains largely undiscovered. Here, we used the sublethal doses of indoxacarb on the S. invicta collected from the eight different cities of Southern China. The alteration in the transcriptome profile of S. invicta following sublethal dosages of indoxacarb was characterized using high-throughput RNA-seq technology. We created 2 libraries, with 50.93 million and 47.44 million clean reads for indoxacarb treatment and control, respectively. A total of 2018 unigenes were regulated after insecticide treatment. Results indicated that a total of 158 differentially expressed genes (DEGs) were identified in the indoxacarb-treated group, of which 100 were significantly upregulated and 58 were downregulated, mostly belonging to the detoxification enzymes, such as AChE, CarE, and GSTs. Furthermore, results showed that most of these DEGs were found in several KEGG pathways, including steroid biosynthesis, other drug metabolizing enzymes, glycerolipid metabolism, chemical carcinogenesis, drug-metabolizing cytochrome P450, glutathione metabolism, glycerophospholipid metabolism, glycolysis/gluconeogenesis, and metabolism of xenobiotics. Together, these findings indicated that indoxacarb causes significant alteration in the transcriptome profile and signaling pathways of S. invicta, providing a foundation for further molecular inquiry.

Insights into insecticide-resistance mechanisms in invasive species: Challenges and control strategies.

Threatening the global community is a wide variety of potential threats, most notably invasive pest species. Invasive pest species are non-native organisms that humans have either accidentally or intentionally spread to new regions. One of the most effective and first lines of control strategies for controlling pests is the application of insecticides. These toxic chemicals are employed to get rid of pests, but they pose great risks to people, animals, and plants. Pesticides are heavily used in managing invasive pests in the current era. Due to the overuse of synthetic chemicals, numerous invasive species have already developed resistance. The resistance development is the main reason for the failure to manage the invasive species. Developing pesticide resistance management techniques necessitates a thorough understanding of the mechanisms through which insects acquire insecticide resistance. Insects use a variety of behavioral, biochemical, physiological, genetic, and metabolic methods to deal with toxic chemicals, which can lead to resistance through continuous overexpression of detoxifying enzymes. An overabundance of enzymes causes metabolic resistance, detoxifying pesticides and rendering them ineffective against pests. A key factor in the development of metabolic resistance is the amplification of certain metabolic enzymes, specifically esterases, Glutathione S-transferase, Cytochromes p450 monooxygenase, and hydrolyses. Additionally, insect guts offer unique habitats for microbial colonization, and gut bacteria may serve their hosts a variety of useful services. Most importantly, the detoxification of insecticides leads to resistance development. The complete knowledge of invasive pest species and their mechanisms of resistance development could be very helpful in coping with the challenges and effectively developing effective strategies for the control of invasive species. Integrated Pest Management is particularly effective at lowering the risk of chemical and environmental contaminants and the resulting health issues, and it may also offer the most effective ways to control insect pests.

Effectiveness of Entomopathogenic Fungi on Immature Stages and Feeding Performance of Fall Armyworm, Spodoptera frugiperda (Lepidoptera: Noctuidae) Larvae.

Maize is a major staple crop in China, and the sustainable productivity of this primary crop has been recently threatened by fall armyworm (FAW), Spodoptera frugiperda, invasion. The five fungal isolates, Aspergillus sp. BM-3 and SE-2-1, Cladosporium tenuissimum SE-10, Penicillium citrinum CTD-24, and Beauveria bassiana ZK-5 were assessed for their efficacy in causing mortality against first to sixth instar eggs and neonate larvae seven days post-treatment, and their effects on the feeding performance of sixth instar S. frugiperda larvae at 48 h post-treatment at three concentrations (1 × 106, 1 × 107, and 1 × 108 conidia mL-1) were also assessed. The six instar S. frugiperda larvae were not susceptible to the five tested fungal isolates. However, B. bassiana ZK-5 caused the highest egg mortality of 40, 70, and 85.6% at 1 × 106, 1 × 107, and 1 × 108 conidia mL-1, respectively, followed by P. citrinum CTD-24 (30.6, 50, and 75.6%) and C. tenuissimum SE-10 (25.6, 40, and 55.6%). In addition, B. bassiana ZK-5 caused the highest neonate mortality of 54.3% at 1 × 108 conidia mL-1. B. bassiana ZK-5 and P. citrinum CTD-24 caused cumulative mortality, including 93.3 and 83.3% mortality of eggs and neonates, respectively, at 1 × 108 conidia mL-1. Furthermore, B. bassiana ZK-5 reduced the feeding efficacy of first to third instar S. frugiperda larvae by 66.7 to 78.6%, while P. citrinum CTD-24 and C. tenuissimum SE-10 reduced larval feeding by 48.3 to 57.1% at 1 × 108 conidia mL-1. However, these fungal isolates were less potent in reducing the feeding activity of fourth to sixth instar S. frugiperda larvae (>46% with B. bassiana at 48 h post-treatment). The tested fungal isolates could play an essential role as microbial biopesticides in suppressing the S. frugiperda population in China after further investigations on their efficacy are obtained in the field.

Model Application of Entomopathogenic Fungi as Alternatives to Chemical Pesticides: Prospects, Challenges, and Insights for Next-Generation Sustainable Agriculture.

In the past few decades, the control of pests and diseases of cultivated plants using natural and biological measures has drawn increasing attention in the quest to reduce the level of dependence on chemical products for agricultural production. The use of living organisms, predators, parasitoids, and microorganisms, such as viruses, bacteria, and fungi, has proven to be a viable and sustainable pest management technique. Among the aforementioned, fungi, most importantly the insect-pathogenic species, have been in use for more than 150years. These include the most popular strains belonging to the genera Beauveria, Metarhizium, Isaria, Hirsutella, and Lecanicillium. Their application is usually through an inundative approach, which inherently involves exposure of the fungal spores to unfavorable humidity, temperature, and solar radiation conditions. These abiotic factors reduce the persistence and efficacy of these insect-pathogenic fungi. Despite these limitations, over 170 strains have been formulated as mycopesticides and are available for commercial use. In the last few decades, numerous studies have suggested that these species of entomopathogenic fungi (EPF) offer far more benefits and have broader ecological functions than hitherto presumed. For instance, aside from their roles as insect killers, it has been well established that they also colonize various host plants and, hence, provide other benefits including plant pathogen antagonism and plant growth promotion and serve as sources of novel bioactive compounds and secondary metabolites, etc. In this light, the potential of EPF as alternatives or perhaps as supplements to chemical pesticides in plant protection is discussed in this review. The paper highlights the numerous benefits associated with endophytic fungal entomopathogen and host plant associations, the mechanisms involved in mediating plant defense against pests and pathogens, and the general limitations to the use of EPF in plant protection. A deeper understanding of these plant host-fungus-insect relationships could help unveil the hidden potentials of fungal endophytes, which would consequently increase the level of acceptance and adoption by users as an integral part of pest management programs and as a suitable alternative to chemical inputs toward sustainable crop production.

General Limitations to Endophytic Entomopathogenic Fungi Use as Plant Growth Promoters, Pests and Pathogens Biocontrol Agents.

The multiple roles of fungal entomopathogens in host plants' growth promotion, pest and pathogen management have drawn huge attention for investigation. Endophytic species are known to influence various activities of their associated host plants, and the endophyte-colonized plants have been demonstrated to gain huge benefits from these symbiotic associations. The potential application of fungal endophytes as alternative to inorganic fertilizers for crop improvement has often been proposed. Similarly, various strains of insect pathogenic fungi have been formulated for use as mycopesticides and have been suggested as long-term replacement for the synthetic pesticides that are commonly in use. The numerous concerns about the negative effects of synthetic chemical pesticides have also driven attention towards developing eco-friendly pest management techniques. However, several factors have been underlined to be militating the successful adoption of entomopathogenic fungi and fungal endophytes as plant promoting, pests and diseases control bio-agents. The difficulties in isolation and characterization of novel strains, negative effects of geographical location, vegetation type and human disturbance on fungal entomopathogens, are among the numerous setbacks that have been documented. Although, the latest advances in biotechnology and microbial studies have provided means of overcoming many of these problems. For instance, studies have suggested measures for mitigating the negative effects of biotic and abiotic stressors on entomopathogenic fungi in inundative application on the field, or when applied in the form of fungal endophytes. In spite of these efforts, more studies are needed to be done to achieve the goal of improving the overall effectiveness and increase in the level of acceptance of entomopathogenic fungi and their products as an integral part of the integrated pest management programs, as well as potential adoption as an alternative to inorganic fertilizers and pesticides.

Impact of invasive ant species on native fauna across similar habitats under global environmental changes.

Biotic invasions can predominantly alter the dynamics, composition, functions, and structure of natural ecosystems. Social insects, particularly ants, are among the most damaging invasive alien species. Invasive ant species are among the supreme threats to ecosystems. There are about 23 species of invasive ants recorded worldwide, according to the ant invasive databases. The ecological impacts of invasive ants comprise predation, hybridization, and competition with native species that changes the ecosystem processes with the biodiversity loss and upsurge of pests. The effects of invasion on native fauna in the same habitats might be catastrophic for the native community through various ecological mechanisms, e.g., habitat disturbance, resource competition, limiting the foraging activity of native species, and various other indirect mechanisms of invasive species. Invasive species may have harmful impacts on habitats and devastating effects on natural flora and fauna, and stopping these new species from being introduced is the most effective way to deter future invasions and maintain biodiversity. This paper reviews the literature to evaluate the effects of invasive ant species on the native species, including vertebrates, invertebrates, and plants sharing the same habitats as the non-native species under global environmental changes. We also highlighted the various management strategies that could be adopted in minimizing the adverse effects of these invasive ant species on the natural ecosystem. To this end, strategies that could regulate the mode and rate of invasion by these alien ant species are the most effective ways to deter future invasions and maintain biodiversity.

Temperature-Dependent Biological Control Effectiveness of Tamarixia radiata (Hymenoptera: Eulophidea) Under Laboratory Conditions.

The parasitism rate and host-feeding rate of Tamarixia radiata (Hymenoptera: Eulophidae), an ectoparasitoid of Diaphorina citri (Hemiptera: Liviidae), were evaluated at 20, 27.5, 30, and 35°C, at 70 ± 5% RH, and 14 h of photoperiod. The biological control efficacy of T. radiata was evaluated by linking the age-stage predation rate with the two-sex life table. The net host-feeding rate (C0) by T. radiata was 32.05, 54.40, 17.25, and 1.92 nymphs per female parasitoid at 20, 27.5, 30, and 35°C, respectively. The total net nymphs killing rate (Z0) was 103.02, 223.82, 72.95, and 6.60 nymphs per female parasitoid at 20, 27.5, 30, and 35°C, respectively. Noneffective parasitism rate was observed at 35°C because of high mortality at this temperature. Our results indicated that temperature had meaningful effects on parasitism and host-feeding rate parameters in the laboratory, and may affect biological control efficiency of the parasitoid in the field. The highest host-feeding rate and total biological control efficiency of T. radiata were recorded at 27.5°C. Most importantly, we found that host-feeding activity of the parasitoid is temperature-dependent, and changed across temperature regimes: the host-feeding rate increased as the temperature increased up to 30°C, started to decrease after this temperature and declined to its minimum level at 35°C. This information is valuable for developing biological control and integrated pest management techniques for Asian citrus psyllid management.

Characterization of mycotoxins from entomopathogenic fungi (Cordyceps fumosorosea) and their toxic effects to the development of asian citrus psyllid reared on healthy and diseased citrus plants.

Entomopathogenic fungi (EPF) produce multiple mycotoxins, which play an essential role in improving fungal pathogenesis and virulence. To characterize various mycotoxins from the crude methanol extract of Cordyceps fumosorosea, a major EPF against various insect pests, we performed ultra-performance liquid chromatography coupled to quadrupole time of flight mass spectrometer (UPLC-QTOF MS) technique, and all compounds were identified through molecular mass and formulae. Bassianolide was assessed against the nymphs and adults of Diaphorina citri reared on healthy and Huánglóngbìng (HLB)-diseased Citrus spp. Plants under laboratory conditions. Overall, 17 compounds were identified from the fungal extract and categorized into three groups, i.e. (1) alkaloids (Isariotins A-C), (2) peptides (Bassianolide, Beauverolides, Beauvericin A, Isaridins and Destruxin E) and (3) polyketide (Tenuipyrone). The detected beauverolides (B, C, F, I, Ja) from C. fumosorosea were novel mycotoxins, and their detection intensity was the highest in the fungal extract. Furthermore, bassianolide caused more than 70% and 80% mortality of D. citri nymphs and adults after two days of application, respectively. After three days of chemical application, all nymphal and adult populations of D. citri were killed by bassianolide. However, the mortality rates of both populations, nymphs and adults, were higher on HLB-diseased plants as compared to healthy plants.

Effects of Seedling Age on Colonization Patterns of Citrus limon Plants by Endophytic Beauveria bassiana and Metarhizium anisopliae and Their Influence on Seedlings Growth.

The inoculation methods, the fungal strains, and several other factors are known to influent the success of fungal entomopathogens colonization in plants. The physiological status of the plant could also be another determinant. In the present study, the ability of three strains of Beauveria bassiana and one strain of Metarhizium anisopliae to successfully colonize Citrus limon plants and the influence of seedling age on endophytic colonization success was examined. Three, 4, and 6 months old seedlings were inoculated with 10 mL of 1 × 108 conidial·mL-1 suspensions of each of the four fungal strains via foliar spraying. All fungal strains successfully colonized citrus seedlings and were sustained up to 2 months in colonized plants irrespective of the seedling age, with differences in the mean percentage colonization recorded at various post-inoculation periods among the fungal strains. The highest percent endophytic fungi recovery rate was recorded in the 3 months old seedlings, where fungal mycelia of inoculated fungi were successfully re-isolated from 65.6% of the untreated newly developed leaf and stem tissues. One strain of B. bassiana, BB Fafu-12, significantly improved seedling height and leaf number. The study demonstrates the influence of seedling age on B. bassiana and M. anisopliae successful colonization in the citrus plant.

Temperature-Dependent Demography and Population Projection of Tamarixia radiata (Hymenoptera: Eulophidea) reared on Diaphorina citri (Hemiptera: Liviidae).

Tamarixia radiata Waterson (Hymenoptera: Eulophidea) is the principal natural enemy used for the control of the major citrus pest Diaphorina citri Kuwayama (Hemiptera: Liviidae). In this study, we collected the life-history data of T. radiata at four different temperatures (20, 27.5, 30, and 35°C) and analyzed them by using the age-stage, two-sex life table. The longest preadult developmental time (16.53 d) was observed at 20°C, whereas the shortest one was 7.29 d at 35°C. The preadult development rate was well-fitted to a linear equation. The lower threshold temperature (T0) was 7.85°C and the thermal summation (K) was 193.36 degree-day. The highest fecundity (F) was 322.7 eggs per female was at 27.5°C, whereas the lowest one was 10.8 eggs per female at 35°C. The net reproductive rate (R0) were 70.97, 169.42, 55.70, and 3.25 offspring at 20, 27.5, 30, and 35°C, respectively; the intrinsic rate of increase (r) were 0.1401, 0.3167, 0.3517, and 0.1143 d-1, respectively. The highest values of fecundity, net reproductive rate, intrinsic rate of increase, and finite rate of increase were observed at 27.5°C. The relationships among F, R0, Nf, and N in all treatments were consistent with R0=F×(Nf/N). The age of peak reproductive value was close to the total preoviposition period in all treatments. Population projections based on the age-stage, two-sex life table showed the dynamics of stage structure and its variability. Faster population growth was observed at 27.5 and 30°C.

Endophytic Beauveria bassiana in Foliar-Treated Citrus limon Plants Acting as a Growth Suppressor to Three Successive Generations of Diaphorina citri Kuwayama (Hemiptera: Liviidae).

Entomopathogenic fungi are commonly applied as inundative sprays to protect plants against insect pests. Their artificial establishment as fungal endophytes to provide other benefits to the host plants aside mere protection against the primary pests has also been widely demonstrated. In the present study, two fungal strains of Beauveria bassiana and one strain of Isaria fumosorosea were assessed in a pathogenicity test against adults of Asian citrus psyllid (Diaphorina citri) and found to induce 50% reduction in the survival rate of D. citri adults within 5 days of exposure. The ability of the three fungal strains to endophytically colonize Citrus limon, the impact on plant growth and the effects of systemic colonization on 3 successive generations of D. citri feeding on colonized plants was evaluated. Citrus seedlings at 4 months post-planting were inoculated with each of the fungal strains via foliar spraying. Both strains of B. bassiana successfully colonized the seedlings. One of the B. bassiana strains (BB Fafu-13) was sustained up to 12 weeks in the colonized seedlings, whereas the other B. bassiana strain (BB Fafu-16) was only recovered up to 8 weeks post-inoculation. Isaria fumosorosea (IF Fafu-1) failed to colonize the plant. Both strains of B. bassiana induced significant improvement in plant height and flush production in endophytically colonized seedlings. In addition, endophytic B. bassiana caused 10-15% D. citri adult mortality within 7 days of exposure. Female D. citri feeding on B. bassiana challenged plants laid fewer eggs as compared to those feeding on endophyte-free seedlings, while reduction in adult emergence was recorded on B. bassiana treated plants. With this study, we present the first evidence of B. bassiana artificial establishment as fungal endophyte in citrus plants and its negative effects on D. citri.

Role of Saponins in Plant Defense Against Specialist Herbivores.

The diamondback moth (DBM), Plutella xylostella (Lepidoptera: Plutellidae) is a very destructive crucifer-specialized pest that has resulted in significant crop losses worldwide. DBM is well attracted to glucosinolates (which act as fingerprints and essential for herbivores in host plant recognition) containing crucifers such as wintercress, Barbarea vulgaris (Brassicaceae) despite poor larval survival on it due to high-to-low concentration of saponins and generally to other plants in the genus Barbarea. B. vulgaris build up resistance against DBM and other herbivorous insects using glucosinulates which are used in plant defense. Aside glucosinolates, Barbarea genus also contains triterpenoid saponins, which are toxic to insects and act as feeding deterrents for plant specialist herbivores (such as DBM). Previous studies have found interesting relationship between the host plant and secondary metabolite contents, which indicate that attraction or resistance to specialist herbivore DBM, is due to higher concentrations of glucosinolates and saponins in younger leaves in contrast to the older leaves of Barbarea genus. As a response to this phenomenon, herbivores as DBM has developed a strategy of defense against these plant biochemicals. Because there is a lack of full knowledge in understanding bioactive molecules (such as saponins) role in plant defense against plant herbivores. Thus, in this review, we discuss the role of secondary plant metabolites in plant defense mechanisms against the specialist herbivores. In the future, trials by plant breeders could aim at transferring these bioactive molecules against herbivore to cash crops.