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Title: Consuming Royal Jelly Causes Mosquitoes to Shift Into and Out of Their Overwintering Dormancy
Females of the Northern house mosquito, Culex pipiens, enter an overwintering dormancy, or diapause, in response to short day lengths and low environmental temperatures. Diapausing female mosquitoes feed exclusively on sugar-rich products rather than human or animal blood, thereby reducing disease transmission. During diapause, Major Royal Jelly Protein 1 (MRJP1) is upregulated in females of Cx. pipiens. This protein is highly abundant in royal jelly, a substance produced by honey bees (Apis mellifera), that is fed to future queens throughout larval development and stimulates longevity and fecundity. However, the role of MRJP1 in Cx. pipiens is unknown. We investigated how supplementing the diets of both diapausing and nondiapausing females of Cx. pipiens with royal jelly affects gene expression, egg follicle length, fat content, protein content, longevity, and metabolic profile. We found that feeding royal jelly to long day-reared females significantly reduced the egg follicle lengths of females and switched their metabolic profiles to be similar to diapausing females. In contrast, feeding royal jelly to short day-reared females significantly reduced lifespan and switched their metabolic profile to be similar nondiapausing mosquitoes. Moreover, RNAi directed against MRJPI significantly increased egg follicle length of short day-reared females, suggesting that these females averted diapause, although RNAi against MRJP1 also extended the lifespan of short day-reared females. Taken together, our data show that consuming royal jelly reverses the seasonal responses of Cx. pipiens and that these responses are likely mediated in part by MRJP1.  more » « less
Award ID(s):
1944324
NSF-PAR ID:
10422846
Author(s) / Creator(s):
; ; ;
Date Published:
Journal Name:
bioRxiv
ISSN:
2692-8205
Format(s):
Medium: X
Sponsoring Org:
National Science Foundation
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  1. Introduction

    Females of the Northern house mosquito,Culex pipiens, enter an overwintering dormancy, or diapause, in response to short day lengths and low environmental temperatures that is characterized by small egg follicles and high starvation resistance. During diapause,Culex pipiensMajor Royal Jelly Protein 1 ortholog (CpMRJP1) is upregulated in females ofCx. pipiens. This protein is highly abundant in royal jelly, a substance produced by honey bees (Apis mellifera), that is fed to future queens throughout larval development and induces the queen phenotype (e.g., high reproductive activity and longer lifespan). However, the role of CpMRJP1 inCx. pipiensis unknown.

    Methods

    We first conducted a phylogenetic analysis to determine how the sequence of CpMRJP1 compares with other species. We then investigated how supplementing the diets of both diapausing and nondiapausing females ofCx. pipienswith royal jelly affects egg follicle length, fat content, protein content, starvation resistance, and metabolic profile.

    Results

    We found that feeding royal jelly to females reared in long-day, diapause-averting conditions significantly reduced the egg follicle lengths and switched their metabolic profiles to be similar to diapausing females. In contrast, feeding royal jelly to females reared in short-day, diapause-inducing conditions significantly reduced lifespan and switched their metabolic profile to be similar nondiapausing mosquitoes. Moreover, RNAi directed againstCpMRJPIsignificantly increased egg follicle length of short-day reared females, suggesting that these females averted diapause.

    Discussion

    Taken together, our data show that consuming royal jelly reverses several key seasonal phenotypes ofCx. pipiensand that these responses are likely mediated in part by CpMRJP1.

     
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  3. Abstract

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    Results

    By early October 2021, roughly 95% ofCulex pipienscollected in culverts were in diapause and 98% ofCx. erraticuswere in diapause. Furthermore, gravid and blood-fedCulex salinarius,Cx. pipiens, andCx. restuanswere collected in late November in 2019 and 2021 in standard mosquito traps. In the winter of 2021, the proportions of non-diapausingCulexdecreased within culverts. The last non-diapausingCx. erraticuswas collected in late December 2021 while the final non-diapausingCx. pipienswas collected in mid-January 2022, both in culverts. Roughly 50% ofCx. pipiensterminated diapause by mid-March 2022, further supported by our collections of gravid females in late March in all 3 years of mosquito collection. In fact, male mosquitoes ofCx. pipiens,Cx. restuans, andCx. territanswere collected by the 1st week of May in 2022, indicating that multiple species ofCulexproduced a second generation that reached adulthood by this time.

    Conclusions

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    Graphical Abstract 
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