Delivery of Nucleic Acids through Embryo Microinjection in the Worldwide Agricultural Pest Insect, <em>Ceratitis capitata</em>

The Mediterranean fruit fly (medfly) Ceratitis capitata (Wiedemann) (Diptera: Tephritidae) is a pest species with extremely high agricultural relevance. ...

University of Pavia

In embryos the maternal-to-zygotic transition (MTZ) integrates post-transcriptional regulation of maternal transcripts with transcriptional activation of the zygotic genome. Although the molecular mechanisms underlying this event are being clarified in Drosophila melanogaster, little is know about the embryogenic processes in other insect species. The recent publication of expressed sequence tags (ESTs) from embryos of the global pest species Ceratitis capitata (medfly) has enabled the investigation of embryogenesis in this species and has allowed a comparison of the embryogenic processes in these two related dipteran species, C. capitata and D. melanogaster, that shared a common ancestor 80-100 mya.

Sex and the single embryo: early deveiopment in the Mediterranean fruit fly, Ceratitis capitata.

Diptera have an extraordinary variety of sex determination mechanisms, and Drosophila melanogaster is the paradigm for this group. However, the Drosophila sex determination pathway is only partially conserved and the family Tephritidae affords an interesting example. The tephritid Y chromosome is postulated to be necessary to determine male development. Characterization of Y sequences, apart from elucidating the nature of the male determining factor, is also important to understand the evolutionary history of sex chromosomes within the Tephritidae. We studied the Y sequences from the olive fly, Bactrocera oleae. Its Y chromosome is minute and highly heterochromatic, and displays high heteromorphism with the X chromosome.

Interchromosomal duplications on the Bactrocera oleae Y chromosome imply a distinct evolutionary origin of the sex chromosomes compared to Drosophila.

Glossina fuscipes fuscipes is the main vector of human and animal trypanosomiasis in Africa, particularly in Uganda. Attempts to control/eradicate this species using biological methods require knowledge of its reproductive biology. An important aspect is the number of times a female mates in the wild as this influences the effective population size and may constitute a critical factor in determining the success of control methods. To date, polyandry in G.f. fuscipes has not been investigated in the laboratory or in the wild. Interest in assessing the presence of remating in Ugandan populations is driven by the fact that eradication of this species is at the planning stage in this country.

Polyandry is a common event in wild populations of the Tsetse fly Glossina fuscipes fuscipes and may impact population reduction measures.

&#x6027;&#x6210;&#x719F;&#x548C;&#x5728;&#x6606;&#x866B;&#x4EA4;&#x914D;&#x4E00;&#x822C;&#x9644;&#x6709;&#x4E3B;&#x8981;&#x751F;&#x7406;&#x548C;&#x884C;&#x4E3A;&#x53D8;&#x5316;&#x3002;&#x5F88;&#x591A;&#x8FD9;&#x4E9B;&#x53D8;&#x5316;&#x6D89;&#x53CA;&#x5230;&#x67E5;&#x627E;&#x4E00;&#x4E2A;&#x4F34;&#x4FA3;&#x7684;&#x9700;&#x8981;&#x548C;&#x968F;&#x540E;&#xFF0C;&#x81F3;&#x4E8E;&#x5973;&#x6027;&#xFF0C;&#x4ECE;&#x4EA4;&#x914D;&#x5230;&#x4EA7;&#x5375;&#x884C;&#x4E3A;&#x641C;&#x7D22;&#x5207;&#x6362;&#x3002;&#x5730;&#x4E2D;&#x6D77;&#x5B9E;&#x8747;&#x3001; &#x89D2;&#x819C;&#x7ED3;&#x819C;&#x708E;&#x7ED3;&#x7403;&#x7518;&#x84DD;&#xFF0C;&#x60CA;&#x4EBA;&#x7684;&#x7E41;&#x6B96;&#x80FD;&#x529B;&#x90FD;&#x662F;&#x5BFC;&#x81F4;&#x4E86;&#x5176;&#x4F5C;&#x4E3A;&#x4E00;&#x79CD;&#x5916;&#x6765;&#x6709;&#x5BB3;&#x7269;&#x79CD;&#x7684;&#x6210;&#x529F;&#x7684;&#x56E0;&#x7D20;&#x4E4B;&#x4E00;&#x3002;&#x82E5;&#x8981;&#x6807;&#x8BC6;&#x4E0E;&#x6210;&#x719F;&#x548C;&#x5728;&#x7537;&#x6027;&#x548C;&#x5973;&#x6027;&#x7684;&#x679C;&#x8747;&#x4EA4;&#x914D;&#x72B6;&#x6001;&#x6709;&#x5173;&#x7684;&#x5206;&#x5B50;&#x53D8;&#x5316;&#xFF0C;&#x57FA;&#x4E8E;&#x5FAE;&#x9635;&#x5217;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x65B9;&#x6CD5;&#x7528;&#x4E8E;&#x6BD4;&#x8F83;&#x6027;&#x672A;&#x6210;&#x719F;&#x7684;&#x3001; &#x6210;&#x719F;&#x7684;&#x5904;&#x5973;&#xFF0C;&#x548C;&#x914D;&#x5BF9;&#x4E2A;&#x4EBA;&#x5934; transcriptomes&#x3002;&#x88AB;&#x96C6;&#x4E2D;&#x6CE8;&#x610F;&#x4E30;&#x5BCC;&#x7684;&#x8A8A;&#x672C;&#x6709;&#x5173;&#x751F;&#x6B96;&#x3001; &#x884C;&#x4E3A;&#x3001; &#x611F;&#x5B98;&#x77E5;&#x89C9;&#x7684;&#x5316;&#x5B66;&#x523A;&#x6FC0;&#x548C;&#x514D;&#x75AB;&#x7CFB;&#x7EDF;&#x8FDB;&#x7A0B;&#x4E2D;&#x6240;&#x505A;&#x7684;&#x66F4;&#x6539;&#x3002;&#x5E7F;&#x6CDB;&#x7684;&#x8F6C;&#x5F55;&#x53D8;&#x5316;&#x5F55;&#x671F;&#x95F4;&#x5973;&#x6027;&#x7684;&#x6210;&#x719F;&#xFF0C;&#x800C;&#x76F8;&#x6BD4;&#x4E4B;&#x4E0B;&#xFF0C;&#x5973;&#x6027;&#x7684; post-mating &#x8F6C;&#x5F55;&#x53D8;&#x5316;&#x5219;&#x6E29;&#x548C;&#x3002;&#x5728;&#x96C4;&#x6027;&#x679C;&#x8747;&#xFF0C;&#x8F6C;&#x5F55;&#x7684;&#x53D8;&#x5316;&#x662F;&#x4E00;&#x81F4;&#x7684;&#x6210;&#x719F;&#x8FC7;&#x7A0B;&#x548C;&#x7ED3;&#x679C;&#x4EA4;&#x914D;&#x3002;&#x7279;&#x522B;&#x503C;&#x5F97;&#x6CE8;&#x610F;&#x7684;&#x662F;&#x5DF2;&#x88AB;&#x5F55;&#x5236;&#xFF0C;&#x679C;&#x8747;&#xFF0C;&#x8FD9;&#x53EF;&#x80FD;&#x662F;&#x7531;&#x4E8E;&#x4E0D;&#x540C;&#x7684;&#x751F;&#x6B96;&#x7B56;&#x7565;&#x7684;&#x8FD9;&#x4E9B;&#x7269;&#x79CD;&#x4EA4;&#x914D;&#x8BF1;&#x5BFC;&#x514D;&#x75AB;&#x5E94;&#x7B54;&#x7684;&#x7F3A;&#x4E4F;&#x3002;&#x8FD9;&#x9879;&#x7814;&#x7A76;&#xFF0C;&#x589E;&#x52A0;&#x6211;&#x4EEC;&#x7684;&#x80CC;&#x540E;&#x6210;&#x719F;&#x7684;&#x5206;&#x5B50;&#x673A;&#x5236;&#x7684;&#x7406;&#x89E3;&#x548C;&#x5728;&#x679C;&#x8747;&#x4EA4;&#x914D;&#x5DF2;&#x786E;&#x5B9A;&#x5728;&#x6B64;&#x866B;&#x5BB3;&#x7684;&#x672A;&#x6765;&#x7BA1;&#x7406;&#x53EF;&#x80FD;&#x6709;&#x7528;&#x7684;&#x91CD;&#x8981;&#x57FA;&#x56E0;&#x76EE;&#x6807;&#x3002;

&#x6027;&#x6210;&#x719F;&#x8F6C;&#x5F55;&#x7EC4;&#x5206;&#x6790;&#x548C;&#x7BA1;&#x5728;&#x5730;&#x4E2D;&#x6D77;&#x5B9E;&#x8747;&#x3001; &#x89D2;&#x819C;&#x7ED3;&#x819C;&#x708E;&#x7ED3;&#x7403;&#x7518;&#x84DD;&#x3002;

Sexual maturation and mating in insects are generally accompanied by major physiological and behavioural changes. Many of these changes are related to the need to locate a mate and subsequently, in the case of females, to switch from mate searching to oviposition behaviour. The prodigious reproductive capacity of the Mediterranean fruit fly, Ceratitis capitata, is one of the factors that has led to its success as an invasive pest species. To identify the molecular changes related to maturation and mating status in male and female medfly, a microarray-based gene expression approach was used to compare the head transcriptomes of sexually immature, mature virgin, and mated individuals. Attention was focused on the changes in abundance of transcripts related to reproduction, behaviour, sensory perception of chemical stimulus, and immune system processes. Broad transcriptional changes were recorded during female maturation, while post-mating transcriptional changes in females were, by contrast, modest. In male medfly, transcriptional changes were consistent both during maturation and as a consequence of mating. Of particular note was the lack of the mating-induced immune responses that have been recorded for Drosophila melanogaster, that may be due to the different reproductive strategies of these species. This study, in addition to increasing our understanding of the molecular machinery behind maturation and mating in the medfly, has identified important gene targets that might be useful in the future management of this pest.

Transcriptome Profiling of Sexual Maturation and Mating in the Mediterranean Fruit Fly, Ceratitis capitata.

La maturation sexuelle et l&#x26;#39;accouplement chez les insectes sont g&#xE9;n&#xE9;ralement accompagn&#xE9;s par de grands changements physiologiques et comportementaux. Beaucoup de ces changements sont li&#xE9;s &#xE0; la n&#xE9;cessit&#xE9; de rep&#xE9;rer une femelle et par la suite, dans le cas des femmes, pour passer de la recherche compagnon &#xE0; la ponte des comportements. La prodigieuse capacit&#xE9; de reproduction de la mouche m&#xE9;diterran&#xE9;enne des fruits, Ceratitis capitata, est l&#x26;#39;un des facteurs qui a conduit &#xE0; son succ&#xE8;s comme une esp&#xE8;ce envahissante contre les ravageurs. Pour identifier les modifications mol&#xE9;culaires li&#xE9;s &#xE0; la maturation et l&#x26;#39;&#xE9;tat d&#x26;#39;accouplement dans la mouche des fruits m&#xE2;les et femelles, une approche bas&#xE9; sur un micror&#xE9;seau d&#x26;#39;expression g&#xE9;nique a &#xE9;t&#xE9; utilis&#xE9;e pour comparer les transcriptomes t&#xEA;te de sexuellement immature, vierge mature, et se sont accoupl&#xE9;s individus. L&#x26;#39;attention a &#xE9;t&#xE9; concentr&#xE9;e sur les changements dans l&#x26;#39;abondance des transcrits li&#xE9;s &#xE0; la reproduction, le comportement, la perception sensorielle du stimulus chimique, et les processus du syst&#xE8;me immunitaire. Les grands changements de transcription ont &#xE9;t&#xE9; enregistr&#xE9;es au cours de la maturation des femmes, tandis que l&#x26;#39;apr&#xE8;s-accouplement changements transcriptionnels chez les femmes &#xE9;taient, en revanche, modeste. Dans mouche m&#xE2;le, les changements de transcription &#xE9;taient conformes &#xE0; la fois pendant la maturation et &#xE0; la suite de l&#x26;#39;accouplement. Notons en particulier le manque des r&#xE9;ponses induites par l&#x26;#39;accouplement immunitaires qui ont &#xE9;t&#xE9; enregistr&#xE9;s pour Drosophila melanogaster, qui peut &#xEA;tre due &#xE0; des strat&#xE9;gies diff&#xE9;rentes en mati&#xE8;re de reproduction de ces esp&#xE8;ces. Cette &#xE9;tude, en plus d&#x26;#39;accro&#xEE;tre notre compr&#xE9;hension de la machinerie mol&#xE9;culaire derri&#xE8;re la maturation et l&#x26;#39;accouplement de la mouche m&#xE9;diterran&#xE9;enne, a identifi&#xE9; des cibles de g&#xE8;nes importants qui pourraient &#xEA;tre utiles dans la gestion future de ce ravageur.

Profilage du transcriptome de la maturation sexuelle et l&#x26;#39;accouplement chez la mouche m&#xE9;diterran&#xE9;enne des fruits, Ceratitis capitata

Sexuelle Reifung und Paarung bei Insekten werden im Allgemeinen von physiologischen und Verhaltensst&ouml;rungen Haupt&auml;nderungen begleitet. Viele dieser &Auml;nderungen beziehen sich auf die Notwendigkeit, einen Partner zu finden, und sp&auml;ter, bei Weibchen von Mate suchen zur Eiablage Verhalten zu wechseln. Die ungeheure reproduktive Kapazit&auml;t der mediterranen Fruchtfliege Ceratitis Capitata, ist einer der Faktoren, die zu seinem Erfolg als invasive Pest Arten gef&uuml;hrt hat. Um die molekularen Ver&auml;nderungen, die im Zusammenhang mit Reifung und Paarung Status in der m&auml;nnlichen und weiblichen Medfly zu identifizieren, war ein Microarray-basierte Gen-Ausdruck-Ansatz verwendet, um den Kopf Transcriptomes sexuell unreif, Reife Jungfrau und gestecktem Personen vergleichen. Aufmerksamkeit konzentrierte sich auf die &Auml;nderungen im &Uuml;berfluss der Transkripte mit Bezug zu Fortpflanzung, Verhalten, Sinneswahrnehmung von chemischen Reiz und Immunsystem-Prozessen. Breite transcriptional &Auml;nderungen wurden w&auml;hrend der weibliche Reifung, aufgezeichnet, w&auml;hrend post-mating transkriptionelle &Auml;nderungen bei Frauen dagegen bescheiden waren. In m&auml;nnlichen Medfly waren transkriptionelle &Auml;nderungen konsequent, sowohl w&auml;hrend der Reifung und als Folge der Paarung. Besonders erw&auml;hnenswert war das Fehlen der Paarung-induzierte Immunantworten, die bei Drosophila Melanogaster festgestellt wurden, die aufgrund der unterschiedlichen reproduktiven Strategien dieser Arten sein k&ouml;nnen. Diese Studie, zus&auml;tzlich zu der erh&ouml;hten unser Verst&auml;ndnis der molekularen Maschinerie hinter Reifung und Paarung in der Medfly, hat wichtige gen Ziele identifiziert, die in die zuk&uuml;nftige Verwaltung von dieser Pest n&uuml;tzlich sein k&ouml;nnten.

Transkriptom-Profiling der sexuellen Reifung und Paaren im Mittelmeer Fruchtfliege Mittelmeerfruchtfliege.

Maturazione sessuale e l&#39;accoppiamento negli insetti sono generalmente accompagnate da importanti cambiamenti fisiologici e comportamentali. Molti di questi cambiamenti sono legati alla necessit&agrave; di individuare un compagno e successivamente, nel caso di femmine, per passare dal compagno di ricerca al comportamento della deposizione delle uova. La prodigiosa capacit&agrave; riproduttiva della Mosca mediterranea della frutta, Ceratitis capitata, &egrave; uno dei fattori che ha portato al suo successo come specie invasiva dei parassiti. Per identificare i cambiamenti molecolari correlati alla maturazione e status degli amori in medfly maschile e femminile, un approccio di espressione genica basata su microarray &egrave; stato utilizzato per confrontare la testa transcriptomes di individui sessualmente immaturi, maturi vergini e riproduttive. L&#39;attenzione &egrave; concentrata sui cambiamenti nell&#39;abbondanza delle trascrizioni relative alla riproduzione, comportamento, percezione sensoriale di stimoli chimici e processi del sistema immunitario. Ampie transcriptional modifiche sono state registrate durante la maturazione femminile, mentre post-mating transcriptional cambiamenti nelle femmine erano, al contrario, modesti. Nel maschio medfly, transcriptional cambiamenti erano coerenti durante la maturazione e in conseguenza di accoppiamento. Di particolare nota &egrave; stata la mancanza della risposta immunitaria indotta da corrispondersi che sono state registrate per la Drosophila melanogaster, che pu&ograve; essere dovuto le diverse strategie riproduttive di queste specie. Questo studio, oltre ad aumentare la nostra comprensione di macchine molecolari dietro la maturazione e l&#39;accoppiamento in medfly, ha individuato gli obiettivi importanti gene che potrebbero essere utili nella gestione futura di questo parassita.

Profilatura transcriptome di maturazione sessuale e corrispondersi nella Mosca mediterranea della frutta, Ceratitis capitata.

&#x6606;&#x866B;&#x306E;&#x6027;&#x6210;&#x719F;&#x3068;&#x4EA4;&#x914D;&#x306F;&#x4E00;&#x822C;&#x7684;&#x306B;&#x4E3B;&#x8981;&#x306A;&#x751F;&#x7406;&#x5B66;&#x7684;&#x304A;&#x3088;&#x3073;&#x884C;&#x52D5;&#x306E;&#x5909;&#x5316;&#x3092;&#x4F34;&#x3063;&#x3066;&#x3044;&#x308B;&#x3002;&#x3053;&#x308C;&#x3089;&#x306E;&#x5909;&#x66F4;&#x306E;&#x591A;&#x304F;&#x306F;&#x7523;&#x5375;&#x884C;&#x52D5;&#x306B;&#x691C;&#x7D22;&#x30E1;&#x30A4;&#x30C8;&#x304B;&#x3089;&#x5207;&#x308A;&#x66FF;&#x3048;&#x308B;&#x306B;&#x306F;&#x3001;&#x5973;&#x6027;&#x306E;&#x5834;&#x5408;&#x306B;&#x306F;&#x3001;&#x305D;&#x306E;&#x5F8C;&#x30E1;&#x30A4;&#x30C8;&#x3092;&#x898B;&#x3064;&#x3051;&#x3066;&#x3001;&#x5FC5;&#x8981;&#x6027;&#x306B;&#x95A2;&#x9023;&#x3057;&#x3066;&#x3044;&#x307E;&#x3059;&#x3002;&#x5730;&#x4E2D;&#x6D77;&#x30DF;&#x30D0;&#x30A8;&#x3001;&#x30C1;&#x30C1;&#x30E5;&#x30A6;&#x30AB;&#x30A4;&#x30DF;&#x30D0;&#x30A8;&#x306E;&#x9A5A;&#x7570;&#x7684;&#x306A;&#x751F;&#x6B96;&#x80FD;&#x529B;&#x306F;&#x3001;&#x4FB5;&#x8972;&#x6027;&#x5BB3;&#x866B;&#x7A2E;&#x3068;&#x3057;&#x3066;&#x306E;&#x6210;&#x529F;&#x306B;&#x3064;&#x306A;&#x304C;&#x3063;&#x305F;&#x8981;&#x56E0;&#x306E;&#x4E00;&#x3064;&#x3067;&#x3042;&#x308B;&#x3002;&#x6210;&#x719F;&#x96C4;&#x3068;&#x96CC;medfly&#x306E;&#x4EA4;&#x5C3E;&#x72B6;&#x6CC1;&#x306B;&#x95A2;&#x9023;&#x3057;&#x305F;&#x5206;&#x5B50;&#x306E;&#x5909;&#x5316;&#x3092;&#x8B58;&#x5225;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x3001;&#x30DE;&#x30A4;&#x30AF;&#x30ED;&#x30A2;&#x30EC;&#x30A4;&#x30D9;&#x30FC;&#x30B9;&#x306E;&#x907A;&#x4F1D;&#x5B50;&#x767A;&#x73FE;&#x306E;&#x30A2;&#x30D7;&#x30ED;&#x30FC;&#x30C1;&#x306F;&#x3001;&#x6027;&#x7684;&#x306B;&#x672A;&#x719F;&#x306A;&#x3001;&#x6210;&#x719F;&#x3057;&#x305F;&#x51E6;&#x5973;&#x306E;&#x30D8;&#x30C3;&#x30C9;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30AF;&#x30EA;&#x30D7;&#x30C8;&#x30FC;&#x30E0;&#x3092;&#x6BD4;&#x8F03;&#x3059;&#x308B;&#x305F;&#x3081;&#x306B;&#x4F7F;&#x7528;&#x3055;&#x308C;&#x3001;&#x500B;&#x4EBA;&#x3092;&#x4EA4;&#x914D;&#x3055;&#x305B;&#x305F;&#x3002;&#x6CE8;&#x76EE;&#x306F;&#x3001;&#x518D;&#x751F;&#x306B;&#x95A2;&#x9023;&#x3057;&#x305F;&#x8EE2;&#x5199;&#x7523;&#x7269;&#x306E;&#x5B58;&#x5728;&#x91CF;&#x306E;&#x5909;&#x5316;&#x3001;&#x884C;&#x52D5;&#x3001;&#x5316;&#x5B66;&#x7684;&#x523A;&#x6FC0;&#x306E;&#x77E5;&#x899A;&#x3001;&#x514D;&#x75AB;&#x30B7;&#x30B9;&#x30C6;&#x30E0;&#x306E;&#x30D7;&#x30ED;&#x30BB;&#x30B9;&#x306B;&#x7126;&#x70B9;&#x3092;&#x5F53;&#x3066;&#x307E;&#x3057;&#x305F;&#x3002;&#x96CC;&#x3067;&#x306F;&#x4EA4;&#x914D;&#x5F8C;&#x306E;&#x8EE2;&#x5199;&#x5909;&#x5316;&#x306F;&#x3001;&#x5BFE;&#x7167;&#x7684;&#x306B;&#x3001;&#x63A7;&#x3048;&#x3081;&#x3067;&#x3042;&#x3063;&#x305F;&#x9593;&#x306B;&#x5E83;&#x7BC4;&#x306A;&#x8EE2;&#x5199;&#x306E;&#x5909;&#x66F4;&#x306F;&#x3001;&#x96CC;&#x306E;&#x6210;&#x719F;&#x4E2D;&#x306B;&#x8A18;&#x9332;&#x3055;&#x308C;&#x305F;&#x3002;&#x7537;&#x6027;medfly&#x3067;&#x306F;&#x3001;&#x8EE2;&#x5199;&#x306E;&#x5909;&#x5316;&#x306F;&#x3001;&#x6210;&#x719F;&#x4E2D;&#x304A;&#x3088;&#x3073;&#x4EA4;&#x914D;&#x306E;&#x7D50;&#x679C;&#x3068;&#x3057;&#x3066;&#x4E21;&#x65B9;&#x3068;&#x3082;&#x4E00;&#x81F4;&#x3057;&#x3066;&#x3044;&#x305F;&#x3002;&#x7279;&#x306B;&#x6CE8;&#x76EE;&#x3059;&#x3079;&#x304D;&#x306F;&#x3001;&#x3053;&#x308C;&#x3089;&#x306E;&#x7A2E;&#x306E;&#x7570;&#x306A;&#x308B;&#x7E41;&#x6B96;&#x6226;&#x7565;&#x304C;&#x539F;&#x56E0;&#x3067;&#x3042;&#x308B;&#x53EF;&#x80FD;&#x6027;&#x304C;&#x3042;&#x308A;&#x30AD;&#x30A4;&#x30ED;&#x30B7;&#x30E7;&#x30A6;&#x30B8;&#x30E7;&#x30A6;&#x30D0;&#x30A8;&#x306E;&#x305F;&#x3081;&#x306B;&#x8A18;&#x9332;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x4EA4;&#x914D;&#x306B;&#x3088;&#x3063;&#x3066;&#x8A98;&#x767A;&#x3055;&#x308C;&#x308B;&#x514D;&#x75AB;&#x5FDC;&#x7B54;&#x306E;&#x6B20;&#x5982;&#x304C;&#x3042;&#x3063;&#x305F;&#x3002;&#x672C;&#x7814;&#x7A76;&#x3067;&#x306F;&#x3001;medfly&#x306E;&#x6210;&#x719F;&#x3068;&#x4EA4;&#x914D;&#x306E;&#x80CC;&#x5F8C;&#x306B;&#x3042;&#x308B;&#x5206;&#x5B50;&#x6A5F;&#x69CB;&#x306E;&#x6211;&#x3005;&#x306E;&#x7406;&#x89E3;&#x3092;&#x6DF1;&#x3081;&#x308B;&#x3053;&#x3068;&#x306B;&#x52A0;&#x3048;&#x3066;&#x3001;&#x3053;&#x306E;&#x5BB3;&#x866B;&#x306E;&#x5C06;&#x6765;&#x306E;&#x7BA1;&#x7406;&#x306B;&#x6709;&#x7528;&#x3067;&#x3042;&#x308B;&#x53EF;&#x80FD;&#x6027;&#x306E;&#x3042;&#x308B;&#x91CD;&#x8981;&#x306A;&#x907A;&#x4F1D;&#x5B50;&#x6A19;&#x7684;&#x3092;&#x540C;&#x5B9A;&#x3057;&#x305F;&#x3002;

&#x30C1;&#x30C1;&#x30E5;&#x30A6;&#x30AB;&#x30A4;&#x30DF;&#x30D0;&#x30A8;&#x3001;&#x30C1;&#x30C1;&#x30E5;&#x30A6;&#x30AB;&#x30A4;&#x30DF;&#x30D0;&#x30A8;&#x306E;&#x6027;&#x7684;&#x6210;&#x719F;&#x3068;&#x5D4C;&#x5408;&#x306E;&#x30C8;&#x30E9;&#x30F3;&#x30B9;&#x30AF;&#x30EA;&#x30D7;&#x30C8;&#x30FC;&#x30E0;&#x30D7;&#x30ED;&#x30D5;&#x30A1;&#x30A4;&#x30EA;&#x30F3;&#x30B0;

&#xC131;&#xC801; &#xC131;&#xC219;&#xACFC; &#xACE4;&#xCDA9;&#xC758; &#xC9DD;&#xC9D3;&#xAE30;&#xB294; &#xC77C;&#xBC18;&#xC801;&#xC73C;&#xB85C; &#xC8FC;&#xC694; &#xC0DD;&#xB9AC; &#xBC0F; &#xD589;&#xB3D9; &#xBCC0;&#xD654;&#xC5D0; &#xC758;&#xD574; &#xB3D9;&#xBC18; &#xB429;&#xB2C8;&#xB2E4;. &#xC774;&#xB7EC;&#xD55C; &#xBCC0;&#xD654;&#xC758; &#xB9CE;&#xC740; &#xC9DD;&#xC744; &#xCC3E;&#xC744; &#xD544;&#xC694;&#xAC00; &#xD558; &#xACE0; &#xADF8; &#xD6C4;, &#xC5EC;&#xC131;, &#xC0B0;&#xB780; &#xD589;&#xB3D9;&#xC744; &#xAC80;&#xC0C9; &#xCE5C;&#xAD6C;&#xC5D0;&#xC11C; &#xC804;&#xD658;&#xC758; &#xACBD;&#xC6B0; &#xAD00;&#xB828; &#xB429;&#xB2C8;&#xB2E4;. &#xC9C0;&#xC911;&#xD574; &#xACFC;&#xC77C; &#xD30C;&#xB9AC; Ceratitis capitata&#xC758; &#xB9C9;&#xB300; &#xD55C; &#xC0DD;&#xC2DD; &#xB2A5;&#xB825; &#xCE68;&#xC785; &#xD574;&#xCDA9; &#xC885;&#xC871;&#xC73C;&#xB85C; &#xC131;&#xACF5;&#xC744; &#xC8FC;&#xB3C4;&#xD558;&#xB294; &#xC694;&#xC778; &#xC911; &#xD558;&#xB098;&#xC785;&#xB2C8;&#xB2E4;. &#xC131;&#xC219; &#xD558; &#xACE0; &#xB0A8;&#xC131;&#xACFC; &#xC5EC;&#xC131;&#xC758; medfly &#xC9DD;&#xC9D3;&#xAE30; &#xC0C1;&#xD0DC;&#xC5D0; &#xAD00;&#xB828; &#xB41C; &#xBD84;&#xC790; &#xBCC0;&#xACBD; &#xB0B4;&#xC6A9;&#xC744; &#xC2DD;&#xBCC4;, &#xC720;&#xC804;&#xC790; microarray &#xAE30;&#xBC18; &#xC2DD; &#xC811;&#xADFC; &#xC131;&#xC801;&#xC73C;&#xB85C; &#xBBF8; &#xC131;&#xC219;&#xD55C;, &#xC131;&#xC219;&#xD55C; &#xCC98;&#xB140;, &#xADF8;&#xB9AC;&#xACE0; &#xC131;&#xAD00;&#xACC4; &#xAC1C;&#xC778;&#xC758; &#xBA38;&#xB9AC; transcriptomes &#xBE44;&#xAD50; &#xC0AC;&#xC6A9; &#xB418;&#xC5C8;&#xB2E4;. &#xC8FC;&#xC758; &#xC7AC;&#xC0DD;, &#xD589;&#xB3D9;, &#xAC10;&#xAC01; &#xC9C0; &#xAC01;&#xC758; &#xD654;&#xD559; &#xC790;&#xADF9; &#xBC0F; &#xBA74;&#xC5ED; &#xC2DC;&#xC2A4;&#xD15C; &#xD504;&#xB85C;&#xC138;&#xC2A4;&#xC640; &#xAD00;&#xB828; &#xB41C; &#xC131;&#xC801; &#xC99D;&#xBA85;&#xC11C;&#xC758; &#xD48D;&#xBD80; &#xD558; &#xAC8C;&#xC5D0;&#xC11C; &#xBCC0;&#xD654;&#xC5D0; &#xC9D1;&#xC911; &#xD588;&#xB2E4;. &#xC5EC;&#xC131;&#xC5D0;&#xC11C; post-mating transcriptional &#xBCC0;&#xD654; &#xD588;&#xB2E4; &#xBC18;&#xBA74;, &#xACB8;&#xC190; &#xD55C; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xAD11;&#xBC94;&#xC704; &#xD55C; transcriptional &#xBCC0;&#xD654; &#xC5EC;&#xC131;&#xC758; &#xC131;&#xC219; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xAE30;&#xB85D; &#xB418;&#xC5C8;&#xB2E4;. &#xC131;&#xC219; &#xD558;&#xB294; &#xB3D9;&#xC548; &#xADF8;&#xB9AC;&#xACE0; &#xAD50;&#xBC30;&#xC758; &#xACB0;&#xACFC;&#xB85C; &#xB0A8;&#xC131; medfly transcriptional &#xBCC0;&#xACBD; &#xC77C;&#xAD00; &#xD588;&#xB2E4;. &#xD2B9;&#xD788; Drosophila melanogaster &#xC774;&#xB7EC;&#xD55C; &#xC885;&#xC758; &#xB2E4;&#xB978; &#xC0DD;&#xC2DD; &#xC804;&#xB7B5;&#xC73C;&#xB85C; &#xC778;&#xD574; &#xC788;&#xC744; &#xC218; &#xC788;&#xC2B5;&#xB2C8;&#xB2E4;&#xC5D0; &#xB300; &#xD55C; &#xAE30;&#xB85D; &#xB41C; &#xC9DD;&#xC9D3;&#xAE30; &#xC720;&#xBC1C; &#xB41C; &#xBA74;&#xC5ED; &#xBC18;&#xC751;&#xC758; &#xBD80;&#xC871;&#xC774; &#xC774;&#xC5C8;&#xB2E4;. &#xC774; &#xC5F0;&#xAD6C;&#xB294; &#xC131;&#xC219; &#xB4A4;&#xC5D0; &#xBD84;&#xC790; &#xAE30;&#xACC4;&#xC5D0; &#xB300; &#xD55C; &#xC6B0;&#xB9AC;&#xC758; &#xC774;&#xD574;&#xB294; medfly &#xC9DD;&#xC9D3;&#xAE30; &#xC99D;&#xAC00; &#xC678;&#xC5D0;&#xC774; &#xD574;&#xCDA9;&#xC758; &#xD5A5;&#xD6C4; &#xAD00;&#xB9AC;&#xC5D0; &#xC720;&#xC6A9;&#xD560; &#xC218; &#xC788;&#xB294; &#xC911;&#xC694; &#xD55C; &#xC720;&#xC804;&#xC790; &#xD45C;&#xC801;&#xC744; &#xD655;&#xC778; &#xD588;&#xB2E4;.

&#xC131;&#xC801; &#xC131;&#xC219;&#xC758; Transcriptome &#xD504;&#xB85C; &#xD30C;&#xC77C;&#xB9C1;&#xACFC; Mating Ceratitis capitata &#xC9C0;&#xC911;&#xD574; &#xACFC;&#xC77C; &#xD30C;&#xB9AC;&#xC5D0;.

La maduraci&#xF3;n sexual y el apareamiento de los insectos son generalmente acompa&#xF1;ados por los principales cambios fisiol&#xF3;gicos y de comportamiento. Muchos de estos cambios est&#xE1;n relacionados con la necesidad de ubicar a un compa&#xF1;ero y, posteriormente, en el caso de las mujeres, para cambiar de compa&#xF1;ero en busca de comportamiento de oviposici&#xF3;n. La prodigiosa capacidad reproductiva de la mosca mediterr&#xE1;nea de la fruta, Ceratitis capitata, es uno de los factores que ha contribuido a su &#xE9;xito como una especie de plagas invasoras. Para identificar los cambios moleculares relacionados con la maduraci&#xF3;n y el estado de apareamiento en mosca de la fruta macho y hembra, un enfoque de microarrays de genes basado en la expresi&#xF3;n se utiliza para comparar la transcriptomes la cabeza de la madurez sexual, virgen madura, y se aparearon los individuos. La atenci&#xF3;n se centr&#xF3; en los cambios en la abundancia de las transcripciones relacionadas con la reproducci&#xF3;n, el comportamiento, la percepci&#xF3;n sensorial de los est&#xED;mulos qu&#xED;micos y los procesos del sistema inmunol&#xF3;gico. Amplios cambios transcripcionales se registraron durante la maduraci&#xF3;n hembra, mientras apareamiento post-transcripcional cambios en las hembras estaban, por el contrario, modesto. En moscamed macho, los cambios transcripcionales fueron consistentes tanto durante la maduraci&#xF3;n y como consecuencia de apareamiento. De particular inter&#xE9;s fue la falta de la respuesta inmune inducida por el apareamiento que se han registrado para la Drosophila melanogaster, que puede ser debido a las diferentes estrategias reproductivas de estas especies. Este estudio, adem&#xE1;s de aumentar nuestra comprensi&#xF3;n de la maquinaria molecular detr&#xE1;s de la maduraci&#xF3;n y el apareamiento de la mosca del Mediterr&#xE1;neo, ha identificado los objetivos importantes de genes que podr&#xED;an ser &#xFA;tiles en el futuro manejo de esta plaga.

Transcriptome de perfiles de la maduraci&#xF3;n sexual y el apareamiento de la mosca mediterr&#xE1;nea de la fruta, Ceratitis capitata

Human malaria, a major public health burden in tropical and subtropical countries, is transmitted exclusively by the bite of a female Anopheles mosquito. Malaria control strategies aimed at inducing sexual sterility in natural vector populations are an attractive alternative to the use of insecticides. However, despite their importance as disease vectors, limited information is available on the molecular mechanisms regulating fertility in Anopheles mosquitoes. In the major malaria vector, An. gambiae, the full complement of sperm and seminal fluid required for a female's lifelong egg production is obtained from a single mating event. This single mating has important consequences for the physiology and behavior of An. gambiae females: in particular, they become refractory to further insemination, and they start laying eggs. In other insects including Drosophila, similar post-copulatory changes are induced by seminal proteins secreted by the male accessory glands and transferred to the female during mating. In this review, we analyze the current state of knowledge on the function and characterization of male seminal proteins in An. gambiae, and provide a comparative assessment of the role of these male reproductive factors in other mosquito vectors of human disease in which female post-copulatory behavior has been studied. Knowledge of the factors and mechanisms regulating fertility in An. gambiae and other vectors can help the design of novel control strategies to fight the spread of disease.

Function and composition of male accessory gland secretions in Anopheles gambiae: a comparison with other insect vectors of infectious diseases.

Insect seminal fluid is a complex mixture of proteins, carbohydrates and lipids, produced in the male reproductive tract. This seminal fluid is transferred together with the spermatozoa during mating and induces post-mating changes in the female. Molecular characterization of seminal fluid proteins in the Mediterranean fruit fly, Ceratitis capitata, is limited, although studies suggest that some of these proteins are biologically active.

Transcriptional profiles of mating-responsive genes from testes and male accessory glands of the Mediterranean fruit fly, Ceratitis capitata.

Molecular interactions between male and female factors during mating profoundly affect the reproductive behavior and physiology of female insects. In natural populations of the malaria mosquito Anopheles gambiae, blood-fed females direct nutritional resources towards oogenesis only when inseminated. Here we show that the mating-dependent pathway of egg development in these mosquitoes is regulated by the interaction between the steroid hormone 20-hydroxy-ecdysone (20E) transferred by males during copulation and a female Mating-Induced Stimulator of Oogenesis (MISO) protein. RNAi silencing of MISO abolishes the increase in oogenesis caused by mating in blood-fed females, causes a delay in oocyte development, and impairs the function of male-transferred 20E. Co-immunoprecipitation experiments show that MISO and 20E interact in the female reproductive tract. Moreover MISO expression after mating is induced by 20E via the Ecdysone Receptor, demonstrating a close cooperation between the two factors. Male-transferred 20E therefore acts as a mating signal that females translate into an increased investment in egg development via a MISO-dependent pathway. The identification of this male-female reproductive interaction offers novel opportunities for the control of mosquito populations that transmit malaria.

The interaction between a sexually transferred steroid hormone and a female protein regulates oogenesis in the malaria mosquito Anopheles gambiae.

The Mediterranean fruit fly, Ceratitis capitata (medfly), is an extremely invasive agricultural pest due to its extremely wide host range and its ability to adapt to a broad range of climatic conditions and habitats. Chemosensory behaviour plays an important role in many crucial stages in the life of this insect, such as the detection of pheromone cues during mate pursuit and odorants during host plant localisation. Thus, the analysis of the chemosensory gene repertoire is an important step for the interpretation of the biology of this species and consequently its invasive potential. Moreover, these genes may represent ideal targets for the development of novel, effective control methods and pest population monitoring systems. Expressed sequence tag libraries from C. capitata adult heads, embryos, male accessory glands and testes were screened for sequences encoding putative odorant binding proteins (OBPs). A total of seventeen putative OBP transcripts were identified, corresponding to 13 Classic, three Minus-C and one Plus-C subfamily OBPs. The tissue distributions of the OBP transcripts were assessed by RT-PCR and a subset of five genes with predicted proteins sharing high sequence similarities and close phylogenetic affinities to Drosophila melanogaster pheromone binding protein related proteins (PBPRPs) were characterised in greater detail. Real Time quantitative PCR was used to assess the effects of maturation, mating and time of day on the transcript abundances of the putative PBPRP genes in the principal olfactory organs, the antennae, in males and females. The results of the present study have facilitated the annotation of OBP genes in the recently released medfly genome sequence and represent a significant contribution to the characterisation of the medfly chemosensory repertoire. The identification of these medfly OBPs/PBPRPs permitted evolutionary and functional comparisons with homologous sequences from other tephritids of the genera Bactrocera and Rhagoletis.

Sniffing out chemosensory genes from the Mediterranean fruit fly, Ceratitis capitata.

Anopheles gambiae mosquitoes are major African vectors of malaria, a disease that kills more than 600,000 people every year. Given the spread of insecticide resistance in natural mosquito populations, alternative vector control strategies aimed at reducing the reproductive success of mosquitoes are being promoted. Unlike many other insects, An. gambiae females mate a single time in their lives and must use sperm stored in the sperm storage organ, the spermatheca, to fertilize a lifetime's supply of eggs. Maintenance of sperm viability during storage is therefore crucial to the reproductive capacity of these mosquitoes. However, to date, no information is available on the factors and mechanisms ensuring sperm functionality in the spermatheca. Here we identify cellular components and molecular mechanisms used by An. gambiae females to maximize their fertility. Pathways of energy metabolism, cellular transport, and oxidative stress are strongly regulated by mating in the spermatheca. We identify the mating-induced heme peroxidase (HPX) 15 as an important factor in long-term fertility, and demonstrate that its function is required during multiple gonotrophic cycles. We find that HPX15 induction is regulated by sexually transferred 20-hydroxy-ecdysone (20E), a steroid hormone that is produced by the male accessory glands and transferred during copulation, and that expression of this peroxidase is mediated via the 20E nuclear receptor. To our knowledge, our findings provide the first evidence of the mechanisms regulating fertility in Anopheles, and identify HPX15 as a target for vector control.

Mating activates the heme peroxidase HPX15 in the sperm storage organ to ensure fertility in Anopheles gambiae.

Female insects generally mate multiple times during their lives. A notable exception is the female malaria mosquito Anopheles gambiae, which after sex loses her susceptibility to further copulation. Sex in this species also renders females competent to lay eggs developed after blood feeding. Despite intense research efforts, the identity of the molecular triggers that cause the postmating switch in females, inducing a permanent refractoriness to further mating and triggering egg-laying, remains elusive. Here we show that the male-transferred steroid hormone 20-hydroxyecdysone (20E) is a key regulator of monandry and oviposition in An. gambiae. When sexual transfer of 20E is impaired by partial inactivation of the hormone and inhibition of its biosynthesis in males, oviposition and refractoriness to further mating in the female are strongly reduced. Conversely, mimicking sexual delivery by injecting 20E into virgin females switches them to an artificial mated status, triggering egg-laying and reducing susceptibility to copulation. Sexual transfer of 20E appears to incapacitate females physically from receiving seminal fluids by a second male. Comparative analysis of microarray data from females after mating and after 20E treatment indicates that 20E-regulated molecular pathways likely are implicated in the postmating switch, including cytoskeleton and musculature-associated genes that may render the atrium impenetrable to additional mates. By revealing signals and pathways shaping key processes in the An. gambiae reproductive biology, our data offer new opportunities for the control of natural populations of malaria vectors.

Sexual transfer of the steroid hormone 20E induces the postmating switch in Anopheles gambiae.

Recent advances in genetic engineering are bringing new promise for controlling mosquito populations that transmit deadly pathogens. Here we discuss past and current efforts to engineer mosquito strains that are refractory to disease transmission or are suitable for suppressing wild disease-transmitting populations.

Engineering the control of mosquito-borne infectious diseases.

In the Mediterranean fruit fly (medfly), Ceratitis capitata, a highly invasive agricultural pest species, polyandry, associated with sperm precedence, is a recurrent behaviour in the wild. The absence of tools for the unambiguous discrimination between competing sperm from different males in the complex female reproductive tract has strongly limited the understanding of mechanisms controlling sperm dynamics and use.

Polyandry in the medfly - shifts in paternity mediated by sperm stratification and mixing.

The highly invasive agricultural insect pest Ceratitis capitata (Diptera: Tephritidae) is the most thoroughly studied tephritid fruit fly at the genetic and molecular levels. It has become a model for the analysis of fruit fly invasions and for the development of area-wide integrated pest management (AW-IPM) programmes based on the environmentally-friendly Sterile Insect Technique (SIT). Extensive transcriptome resources and the recently released genome sequence are making it possible to unravel several aspects of the medfly reproductive biology and behaviour, opening new opportunities for comparative genomics and barcoding for species identification. New genes, promotors and regulatory sequences are becoming available for the development/improvement of highly competitive sexing strains, for the monitoring of sterile males released in the field and for determining the mating status of wild females. The tools developed in this species have been transferred to other tephritids that are also the subject of SIT programmes.

Paolo Gabrieli

Department of Biology and Biotechnology

University of Pavia

Delivery of Nucleic Acids through Embryo Microinjection in the Worldwide Agricultural Pest Insect, Ceratitis capitata

Paolo Gabrieli

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Biology and Biotechnology

University of Pavia

Delivery of Nucleic Acids through Embryo Microinjection in the Worldwide Agricultural Pest Insect, Ceratitis capitata

Department of Biology and Biotechnology