First record of Fusarium concentricum (Hypocreales: Hypocreaceae) isolated from the moth Polychrosis cunninhamiacola (Lepidoptera: Tortricidae) as an entomopathogenic fungus.

Fusarium concentricum Nirenberg & O' Donnell (Ascomycota: Hypocreales) is a fungal species known to infect plants, but never reported as entomopathogenic. Polychrosis cunninhamiacola Liu et Pei (Lepidoptera: Tortricidae: Olethreutinae) is a major and widespread insect pest causing economic losses to cultivated Chinese fir Cunninghamia lanceolata (Lamb.) Hook. It is routinely controlled by extensive use of chemical insecticides, which is perceived as environmentally unsustainable. During March and April of 2019-2020, muscardine cadavers of larvae and pupae of P. cunninhamiacola infected with growing fungus were collected in a fir forest in northern Guangdong Province, China. Conidia were isolated and cultured on PDA medium, from which the fungal strain was identified as F. concentricum FCPC-L01 by morphology and by sequence alignment match with Tef-1α gene. Pathogenicity bioassays at the conidial concentration 1 × 107 revealed P. cunninhamiacola adults and Danaus chrysippus (L.) (Lepidoptera: Nymphalidae) larvae are sensitive to the fungal infection, but not the fire ant Solenopsis invicta Buren (Hymenoptera: Formicidae). We believe results indicate this fungal strain might be applicable against specific target insect pests. As this is the first record of a natural infection caused by F. concentricum in insects, we propose host specificity tests should be done to evaluate its potential as a biocontrol agent.

Metarhizium Entomopathogenic Fungi Against the Beetle Brontispa longissima (Coleoptera: Hispidae): Isolation and Species Identification.

Seven entomopathogenic fungi strains (M1-7) were isolated from field-obtained dead coconut hispine beetles Brontispa longissima (Gestro), identified to species, and bioassayed for their pathogenicity. According to ITS sequences, all isolates belong in the genus Metarhizium, mainly M. flavoviride and M. anisopliae. Measured median lethal times (LT50) of 1×107 conidia/ml of M1-7 against fourth-instar B. longissima larvae within 15 d following exposure were, respectively: 5.43, 10.64, 11.26, 10.93, 6.62, 4.73, and 5.95 d. The isolate M6 yielded the highest mortality to fourth-instar larvae, and was thus selected to be tested against other larval instars and adults of B. longissima, after Time-Dose-Mortality (TDM) models. M6 proved more pathogenic against larvae than adults. The obtained bioassays data produced a good fit to the TDM models, yielding estimated LC50 and LT50 for each of the tested developmental stages of B. longissima. Both the obtained dose (ß) and time effect (ri) parameters from TDM models suggest that first-instar larvae are the most susceptible life stage of the pest insect, while adults are more resistant to M6 infection. Calculated LC50 values were, respectively, 1.23×103 and 1.15×106 conidia/ml for first-instar larvae and adults, on the 15th day following M6 inoculation. Estimated LT50 were 3.3 and 5.9 d for first-instar larvae and adults, respectively, at 1×108 conidia/ml. Taken together, these results would suggest Metarhizium M6 as an option for the biological control of B. longissima in the field.

New records of Celoporthe guangdongensis and Cytospora rhizophorae on mangrove apple in China.

BACKGROUND: Sonneratia apetala Francis Buchanan-Hamilton (Sonneratiaceae, Myrtales), is a woody species with high adaptability and seed production capacity. S. apetala is widely cultivated worldwide as the main species for mangrove construction. However, the study of diseases affecting S. apetala is limitted, with only a few fungal pathogens being recorded. Cryphonectriaceae (Diaporthales) species are the main pathogens of plants. They can cause canker diseases to several trees and thereby seriously threaten the health of the hosts. These pathogens include Cryphonectria parasitica (Cryphonectriaceae) causing chestnut blight on Castanea (Rigling and Prospero 2017) and Cytospora chrysosperma (Cytosporaceae) causing polar and willow canker to Populus and Salix (Wang et al. 2015) . Therefore, the timely detection of of Cryphonectriaceae canker pathogens on S. apetala is extremely important for protecting the mangrove forests. NEW INFORMATION: Two diaporthalean fungi, Celoporthe guangdongensis and Cytospora rhizophorae have been reported for the first time to cause canker on the branches of S. apetala. C. guangdongensis is significantly pathogenic and C. rhizophorae is saprophytic on S. apetala.

Differential Behavioral Responses of Solenopsis invicta (Hymenoptera: Formicidae) Workers Toward Nestmate and Non-Nestmate Corpses.

The removal of corpses (aka 'necrophoric behavior') is critical to sanitation in ant colonies. However, little is known about differences in the necrophoric responses of Solenopsis invicta workers towards corpses of nestmates and non-nestmates. We introduced corpses of S. invicta workers from either intracolony (i.e., nestmate) or intercolony (i.e., non-nestmate) origin at the entrance of artificial nests, and recorded workers' aggressive responses and necrophoric behaviors for analysis. Solenopsis invicta workers displayed distinct responses towards corpses of different origins. Specifically, resident workers were more likely to remove fresh non-nestmate corpses than nestmate corpses, but there was no difference regarding corpses that had been dead for 15 min or longer. Resident workers reacted more aggressively to, and removed more quickly, fresh non-nestmate corpses than corpses of their nestmates. On the other hand, there was no significant difference in the removal time between nestmate and non-nestmate corpses that had been dead for 15 min or longer. Resident workers always displayed stronger aggressiveness towards non-nestmate corpses than nestmate corpses, excepting to corpses that had been dead for 6 h, which elicited a response. No significant correlation between the removal times and aggressiveness levels were detected in any treatments. It remains to be tested whether this differential response is adaptive in how it influences colony fitness and competition.

A Chemosensory Protein Gene Si-CSP1 Associated With Necrophoric Behavior in Red Imported Fire Ants (Hymenoptera: Formicidae).

Necrophoric behavior is essential to colony health in social insects. Little is known about the genes that are responsible for necrophoric behavior. Here, we show that a chemosensory protein gene Si-CSP1 was expressed significantly higher in the antennae than in other tissues such as the legs and heads of Solenopsis invicta Buren workers. Furthermore, Si-CSP1-silenced workers moved significantly fewer corpses of their nestmates than normal workers. Finally, Si-CSP1-silenced workers exhibited weaker antennal responses to oleic acid and linoleic acid than controls. These results suggest that Si-CSP1 functions by sensing oleic acid and linoleic acid associated with dead colony members and regulating the necrophoric behavior of workers in S. invicta.

On the Molecular Basis of Division of Labor in Solenopsis invicta (Hymenoptera: Formicidae) Workers: RNA-seq Analysis.

The fire ant Solenopsis invicta Buren is an important invasive pest. Among S. invicta workers behavioral changes depend on age where younger ants are nurses and older ants foragers. To identify potential genes associated with this division of labor, we compared gene expression between foragers and nurses by high-throughput sequencing. In total, we identified 1,618 genes significantly differently expressed between nurses and foragers, of which 542 were upregulated in foragers and 1,076 were upregulated in nurses. Several pathways related to metabolism were significantly enriched, such as lipid storage and fatty acid biosynthesis, which might contribute to the division of labor in S. invicta. Several genes involved in DNA methylation, transcription, and olfactory responses as well as resistance to stress were differentially expressed between nurses and foragers workers. Finally, a comparison between previously published microarray data and our RNA-seq data in S. invicta shows 116 genes overlap, and the GO term myofibril assembly (GO: 0030239) were simultaneously significantly enriched. These results advance knowledge of potentially important genes and molecular pathways associated with worker division of labor in S. invicta. We hope our dataset will provide . candidate target genes to disrupt organization in S. invicta as a control strategy against this invasive pest.

Metarhizium anisopliae infection alters feeding and trophallactic behavior in the ant Solenopsis invicta.

In social insects, social behavior may be changed in a way that preventing the spread of pathogens. We infected workers of the ant Solenopsis invicta with an entomopathogenic fungus Metarhizium anisopliae and then videotaped and/or measured worker feeding and trophallactic behavior. Results showed that fungal infected S. invicta enhanced their preference for bitter alkaloid chemical quinine on 3days after inoculation, which might be self-medication of S. invicta by ingesting more alkaloid substances in response to pathogenic infection. Furthermore, infected ants devoted more time to trophallactic behavior with their nestmates on 3days post inoculation, in return receiving more food. Increased interactions between exposed ants and their naive nestmates suggest the existence of social immunity in S. invicta. Overall, our study indicates that S. invicta may use behavioral defenses such as self-medication and social immunity in response to a M. anisopliae infection.

[Bahaviour of Solenopsis invicta workers to protect pupae from infection by Metarhizium anisopliae].

Previous studies have focused on how ants deal with workers infected by pathogens but how pupae are protected from infection by fungi is not well understood. The behavioral mechanisms adopted by Solenopsis invicta (red imported fire ants, RIFA) adult workers to protect pupae against Metarhizium anisopliae infection were studied. We observed the behavioral changes of M. anisopliae infected adult workers in the brood chamber as well as the behavioral changes of healthy workers to fungus exposed pupae. The time of fungus infected workers spent in the pupal chamber reduced significantly from 103.4 s on the first day to 38.5 s on the third day. Moreover, the percentage of time spending on brood care in the pupal chamber reduced significantly from 13.6% on the first day to 3.5% on the third day. When pupae were infected by M. anisopliae, workers performed 5.3 times more grooming to fungus exposed pupae than controls, and the duration of each grooming bout to fungus exposed pupae was 5.2 times longer than controls. Grooming did remove many conidia on the surface of fungus exposed pupae. The mean numbers of conidia on the surface of pupae were 103.1, 51.6 and 31.3 when no workers, two workers and ten workers accompanied a pupa, respectively. The presence of workers resulted in a lower germination rate of conidia on the surface of pupae. The mean germination rates of conidia after 20 h of inoculation on the surface of pupae were 95.1%, 80.4% and 59.9%, in the treatments with no worker, two workers and ten workers respectively. There was a positive correlation between the emergence rate of pupae and the number of accompanying workers. RIFA protect their pupae from infection by M. anisopliae through social be- haviors which enable the sustainable development of their population.