Isolation of Beauveria Strains and Their Potential as Control Agents for Lema bilineata Germar (Coleoptera: Chrysomelidae)

Lema bilineata Germar (Coleoptera: Chrysomelidae) was recently reported to damage Physalis peruviana crops in Brazil. Given the potential for inflicting damage on other Solanaceae species and the lack of alternatives for controlling this pest, we assessed the pathogenicity of 15 Beauveria isolates against L. bilineata adults in vitro. In addition, three of these isolates were tested for their ovicidal effect against L. bilineata eggs. Fungal strains were isolated from mummified corpses of L. bilineata collected in a non-commercial field in Paraná, Brazil. The isolates were identified as Beauveria bassiana using molecular markers. Lema bilineata adults were susceptible to conidial suspensions of all these isolates at a concentration of 108 conidia mL−1. Deaths caused by fungal extrusion were confirmed. Three strains were found to be more virulent against L. bilineata adults and showed ovicidal effects. This is the first study on entomopathogenic fungi isolated from dead insects collected from P. peruviana crops and tested against L. bilineata carried out in Brazil. The results obtained in the laboratory indicate the high potential of the use of three B. bassiana strains against L. bilineata as a biocontrol agent.

Effectiveness of Cordyceps sp. and Beauveria Bassiana Against Large Cabbage-heart Caterpillar, Crocidolomia Pavonana Fabricius (Lepidoptera: Crambidae)

Crocidolomia pavonana Fabr. (Lepidoptera: Crambidae) is a major pest on various cabbage crops in Indonesia. Various insecticides have been used to control this insect pest including several entomopathogenic fungi, and this cabbage caterpillar has become resistant to various insecticides. In order to develop insecticides from entomopathogenic fungi, species or varieties from natural area such as forest are needed. A bioassays of Cordyceps sp. and Beauveria bassiana has been carried out in the laboratory. Inoculum of Cordyceps sp. obtained from the Dipterocarpaceae forest in South Kalimantan, Indonesia recovered from dead ants by the mycelium of Cordyceps sp. Four concentration of Cordyceps sp. and Beauveria bassiana inoculum and tween80 as a control were tested against 3rd instar larvae of C. pavonana with five replications. Field efficacy trial was carried out in a screen house (8 x 12 M2) using Chinese cabbage plants (Brassica juncea) aged 28 days after sowing, with a spacing of 0.4 M between rows. Three formulation of Cordyceps sp. and tween80 as a control were used with 6 replications. The result of the bioassays in the laboratory showed that Cordyceps sp. with concentration of 1 x 107 conidia mL-1 was able to give 100% mortality od C. pavonana larvae, while B. bassiana with the same concentration caused morality of C. pavonana larvae of 98.33%, although statistically not significantly different. The results of probit analysis also showed that LC90 Cordyceps sp. against cabbage caterpillar larvae was 1.57 x 106 conidia mL-1, while B. bassiana required a concentration of 2.6 x 106 conidia mL-1 to reach the LC90 value. The results of filed trial showed that the formulation of Cordyceps sp. 1 x 107 conidia mL-1 using palm oil cooking oil was able to give an average mortality of 86.7%, and significantly higher (P<0.05) than the other treatment. The results of this study indicate that the inoculum of Cordyceps sp. from Dipterocarpaceae forest is very promising to be developed as an insecticide to control C. pavonana in cabbage

A comparative study on the virulence of entomopathogenic fungi against Trogoderma granarium (Everts) (Coleoptera: Dermestidae) in stored grains rice

Entomopathogenic fungi (EPF) now a possible safer microbial control measure that could be considered as a substitute for chemical control of insect pests. Three EPF viz., Metarihizium anisopliae, Isaria furnosoroseus and Beauveria bassiana were evaluated for their virulence against the grubs of Khapra beetle, Trogoderma granarium (Everts) under laboratory conditions. The isolates were applied by two methods viz., diet incorporation and an immersion method with 3rd instar 20 grubs of T. granarium for each. The virulence of EPF was determined using percent mortality. Significantly higher mortality was observed in M. anisopliae applied through immersion (98.33%) and diet incorporation (93.33%) methods followed by B. bassiana (90.83 and 85.83%, respectively). The mortality caused by I. furnosoroseus was statistically lower in immersion and diet incorporation methods i.e. 81.67 and 73.33%, respectively. Based on the immersion method, all EPF were studied for multiple conidial concentration i.e., 1×104, 1×105, 1×106, 1×107 and 1×108 under the same in-vitro conditions. All the isolates were pathogenic to grub of T. granarium at the highest conidial concentration. M. anisopliae was proved the most effective virulent resulting in 98.33% mortality of the pest with LT50 4.61 days at 1 × 108 conidial concentration followed by 90.83 and 81.67 percent mortality with 5.07 and 8.01 days LT50, in the application of B. bassiana and I. furnosoroseus, respectively. M. anisopliae showed higher efficacy and could be considered as promising EPF for the development of myco-insecticides against effective biocontrol of T. granarium.

Root-Associated Entomopathogenic Fungi Modulate Their Host Plant’s Photosystem II Photochemistry and Response to Herbivorous Insects

The escalating food demand and loss to herbivores has led to increasing interest in using resistance-inducing microbes for pest control. Here, we evaluated whether root-inoculation with fungi that are otherwise known as entomopathogens improves tomato (Solanum lycopersicum) leaflets’ reaction to herbivory by Spodoptera exigua (beet armyworm) larvae using chlorophyll fluorescence imaging. Plants were inoculated with Metarhizium brunneum or Beauveria bassiana, and photosystem II reactions were evaluated before and after larval feeding. Before herbivory, the fraction of absorbed light energy used for photochemistry (ΦPSII) was lower in M. brunneum-inoculated than in control plants, but not in B. bassiana-inoculated plants. After herbivory, however, ΦPSII increased in the fungal-inoculated plants compared with that before herbivory, similar to the reaction of control plants. At the same time, the fraction of energy dissipated as heat (ΦNPQ) decreased in the inoculated plants, resulting in an increased fraction of nonregulated energy loss (ΦNO) in M. brunneum. This indicates an increased singlet oxygen (1O2) formation not detected in B. bassiana-inoculated plants, showing that the two entomopathogenic fungi differentially modulate the leaflets’ response to herbivory. Overall, our results show that M. brunneum inoculation had a negative effect on the photosynthetic efficiency before herbivory, while B. bassiana inoculation had no significant effect. However, S. exigua leaf biting activated the same compensatory PSII response mechanism in tomato plants of both fungal-inoculated treatments as in control plants.

Ophiocordyceps aphrophoridarum sp. nov., a new entomopathogenic species from Guizhou, China

Ophiocordyceps is the largest genus in the family Ophiocordicipitaceae, including many entomopathogenic species. In recent years, many species have been described in this genus, with a wide range of host insects. Entomopathogenic fungi include ecologically, economically and medicinally important species, but a large portion of their diversity remains to be discovered and described. In this study, a new species, Ophiocordyceps aphrophoridarum sp. nov, parasitising Aphrophoridae sp. (Hemiptera) is proposed from China, based on evidence from morphology and molecular phylogenetic analyses. This species is characterised by fibrous, pigmented stromata, cylindrical asci and filiform ascospores. Compared to its closest relative, O. tricentri, the new species has wider perithecia and longer asci. Molecular phylogenetic analyses of a multilocus dataset (consisting of SSU, ITS, LSU, TEF1, RPB1 and RPB2) confirm its placement in Ophiocordyceps. Ophiocordyceps aphrophoridarum is morphologically described and illustrated with colour photographs. Morphological comparisons with closely-related species are also presented in tabulated format.

Lobesia botrana: A Biological Control Approach with a Biopesticide Based on Entomopathogenic Fungi in the Winter Season in Chile

Lobesia botrana (Denis and Shiffermüller) (Lepidoptera: Tortricidae) is one of the main pests that affect the production and export of table grapes in Chile. Because this pest has quarantine status, the fruit must be fumigated with methyl bromide, which reduces the fruit’s export competitiveness in the destination market. In the present study, to help resolve this issue, six native entomopathogenic fungi were identified through multilocus analysis, including three Beauveria pseudobassiana and three Metarhizium robertsii. These fungi were evaluated in the laboratory to control L. botrana in its pupal stage in a silk cocoon and compared against a biological control product. Formulations with additional carbon sources improved the performance of the fungi. The treatments with outstanding performance contained the fungal strains B. pseudobassiana RGM 2184 and M. robertsii RGM 678. These strains were evaluated in the field during the winter season in two different regions of the country; the strains reached maximum efficacies of 80% and 88%, respectively, at 21 days post first application. Therefore, entomopathogenic fungi can contribute to reducing pupal populations in winter, thereby decreasing the moth population in spring–summer.

Efficacy of Entomopathogenic Trichoderma Isolates against Sugarcane Woolly Aphid, Ceratovacuna lanigera Zehntner (Hemiptera: Aphididae)

Ceratovacuna lanigera Zehntner (Hemiptera: Aphididae) is a destructive insect pest of sugarcane that is responsible for reducing the quality, yield, and sugar content. Ecofriendly methods of pest control, such as the use of biological control, is encouraged since it does not exhibit the harmful effects that are usually seen when using chemical-based pesticides. Here, we evaluated the efficacy of several entomopathogenic fungi isolates against C. lanigera under laboratory conditions. A total of seven isolates were obtained from agricultural soils that were collected from various locations from Sabah, Malaysia, using the insect bait method. Among the seven, four isolates (TMS114, TMS623, TMS628, and TMS707) were positively identified as T. harzianum (Hypocreales: Hypocreaceae), two isolates (TMS45, TMS321) as T. asperellum (Hypocreales: Hypocreaceae), and one isolate (TMS633) as T. hamatum (Hypocreales: Hypocreaceae) based on a combination of morphological characteristics and molecular data using ITS 1-5.8 S-ITS 2 regions of the rDNA. Four different concentrations (1 × 105, 1 × 106, 1 × 107, and 1 × 108 mL−1) of conidia suspensions of the isolates were applied separately on nymphs and adults of C. lanigera. The results showed that mortality in both nymphs and adults was dependent on the conidia concentration and exposure time. Among the four concentrations, the highest mortality of C. lanigera was found to range from 59.99 to 75.70% for nymphs after 72 h and from 57.45 to 72.31% for adults after 6 days of treatments at the concentration of 1 × 108 conidia mL−1. Trichodermaharzianum (TMS623) showed significantly highest mortality of 75.70% for nymphs and 72.31% for adults among the seven isolates. . However, the nymph and adult mortality rates for the isolates were 70.00% & 68.00% for TMS114, 66.42% & 63.10% for TMS628, 67.85% & 65.24% for TMS707, 61.42% & 58.80% for TMS45, 59.99% & 57.45% for TMS321 and 63.56% & 60.91% for TMS633, respectively. For nymph mortality, T. harzianum (TMS623) showed a LC50 (lethal concentration) value of 6.30 × 105 conidia mL−1 and LC90 value of 3.01 × 109 conidia mL−1, respectively, and for adult mortality, the values were found of 6.16 × 105 conidia mL−1 for LC50 and 2.23 × 1010 conidia mL−1 for LC90, respectively. The lethal time (LT) values for nymph and adult mortality were found of 42.65 h and 3.89 days for LT50 and 93.32 h and 8.70 days for LT90 by T. harzianum (TMS623). Therefore, T. harzianum (TMS623) showed more pathogenic potential against C. lanigera and can be further applied to determine its efficacy under field conditions. To our best knowledge, this is the first report of T.harzianum,T.hamatum, and T.asperellum species that are used as entomopathogenic fungi against sugarcane woolly aphid, C. lanigera.

Evaluation of the Entomopathogenic Potential of Beauveria bassiana, Metarhizium anisopliae and Isaria fumosorosea for Management of Cosmopolites sordidus Germar (Coleoptera: Curculionidae)

The banana weevil (BW), Cosmopolites sordidus, is the main coleopteran pest of banana, causing up to 100% yield loss. In this study, we screened 20 isolates of entomopathogenic fungi (EPF) for the management of BW. In the lab, eight Beauveria bassiana isolates caused >50% mortality of the adult BW, whereas Metarhizium anisopliae and Isaria fumosorosea isolates were less pathogenic. B. bassiana isolates ICIPE 648, ICIPE 660 and ICIPE 273 were the most pathogenic, killing ≥80% of adult BW. B. bassiana isolate ICIPE 622 yielded the highest spores per BW cadaver (1.84 × 108 spores), followed by ICIPE 660, ICIPE 273 and ICIPE 648—1.17 × 108, 3.8 × 107 and 3.6 × 107 spores, respectively. ICIPE 273 had the shortest LT50 (5.3 days) followed by ICIPE 648 (9.8 days) and 660 (11.1 days). Similarly, the LC50 values for the three isolates were 5.18 × 107, 5.49 × 107 and 5.2 × 107 spores mL−1, respectively. In the field, ICIPE 273 and ICIPE 648 had the highest (31.3%) and lowest (20.8%) pathogenicity, respectively. This study indicates that the B. bassiana isolates ICIPE 273, ICIPE 648 and ICIPE 660 are potential candidates for the environmentally sustainable management of BW.