Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree <em>Parasponia andersonii</em>

Parasponia andersonii is a fast-growing tropical tree that belongs to the Cannabis family (Cannabaceae). Together with 4 additional species, it forms the ...

Wageningen University & Research

&#x8C46;&#x79D1;&#x809D;&#x9171; SCR RGA GAI&#xFF09;-&#x952E;&#x5165;&#x8F6C;&#x5F55;&#x56E0;&#x5B50;&#x7ED3;&#x7624;&#x4FE1;&#x53F7; PATHWAY1 (NSP1) &#x548C; NSP2 &#x662F;&#x6839;&#x7624;&#x83CC; Nod &#x56E0;&#x5B50;&#x8BF1;&#x5BFC;&#x7ED3;&#x7624;&#x56FA;&#x6C2E;&#x7684;&#x5FC5;&#x8981;&#x6761;&#x4EF6;&#x3002;&#x8FD9;&#x4E24;&#x79CD;&#x86CB;&#x767D;&#x8D28;&#x88AB;&#x8BA4;&#x4E3A;&#x662F; Nod &#x56E0;&#x5B50;&#x54CD;&#x5E94;&#x56E0;&#x7D20;&#x540E;&#x5171;&#x751F;&#x4FE1;&#x53F7;&#x8C03;&#x8282;&#x57FA;&#x56E0;&#x8868;&#x8FBE;&#x3002;&#x7136;&#x800C;&#xFF0C;&#x8C46;&#x7C7B; NSP1 &#x548C; NSP2 &#x53EF;&#x4EE5;&#x662F;&#x529F;&#x80FD;&#x4E0A;&#x6539;&#x4E3A; nonlegume orthologs&#xFF0C;&#x5305;&#x62EC;&#x5927;&#x7C73; (&#x6C34;&#x7A3B;) NSP1 &#x548C; NSP2&#xFF0C;&#x8868;&#x660E;&#x8FD9;&#x4E24;&#x79CD;&#x86CB;&#x767D;&#x8D28;&#x7684;&#x529F;&#x80FD;&#x4E0A;&#x4FDD;&#x5B88;&#x5728;&#x9AD8;&#x7B49;&#x690D;&#x7269;&#x4E2D;&#x3002;&#x5728;&#x8FD9;&#x91CC;&#xFF0C;&#x6211;&#x4EEC;&#x8868;&#x660E; NSP1 &#x548C; NSP2 strigolactone (SL) &#x751F;&#x7269;&#x5408;&#x6210;&#x5728;&#x8C46;&#x79D1;&#x82DC;&#x84FF;&#x548C;&#x5927;&#x7C73;&#x4E2D;&#x4E0D;&#x53EF;&#x6216;&#x7F3A;&#x3002;&#x7A81;&#x53D8; nsp1 &#x690D;&#x7269;&#x4E0D;&#x4EA7;&#x751F;&#x8865;&#x5145;&#x52B3;&#x5DE5;&#x8BA1;&#x5212;&#xFF0C;&#x800C;&#x5728; M.&#x622A;&#x5F62;&#x82DC;&#x84FF;&#xFF0C;NSP2 &#x662F;&#x5FC5;&#x4E0D;&#x53EF;&#x5C11;&#x7684; orobanchol &#x8F6C;&#x5316;&#x4E3A; didehydro-orobanchol&#xFF0C;&#x8FD9;&#x5C31;&#x662F;&#x7531;&#x8FD9;&#x4E00;&#x7269;&#x79CD;&#x7684;&#x4E3B;&#x8981; SL&#x3002;&#x5728; nsp1 nsp2 &#x7A81;&#x53D8;&#x4F53;&#x80CC;&#x666F;&#x4E2D; SL &#x5408;&#x6210;&#x7684;&#x4E0D;&#x5B89;&#x4E0E;&#x51CF;&#x5C11;&#x8868;&#x8FBE;&#x7684; DWARF27&#xFF0C;SL &#x751F;&#x7269;&#x5408;&#x6210;&#x5173;&#x952E;&#x57FA;&#x56E0;&#x5173;&#x8054;&#x3002;&#x6C34;&#x7A3B;&#x548C; M.&#x622A;&#x5F62;&#x82DC;&#x84FF;&#x4EE3;&#x8868;&#x4E0D;&#x540C;&#x7684;&#x4EB2;&#x7F18;&#x8840;&#x7EDF;&#xFF0C;&#x62C6;&#x5206;&#x5927;&#x7EA6; 1 &#x4EBF; 5000 &#x4E07;&#x591A;&#x5E74;&#x524D;&#x3002;&#x56E0;&#x6B64;&#xFF0C;&#x6211;&#x4EEC;&#x7684;&#x7ED3;&#x8BBA;&#x7531; NSP1 &#x548C; NSP2 &#x7684; SL &#x751F;&#x7269;&#x5408;&#x6210;&#x7684;&#x8C03;&#x63A7;&#x662F;&#x5728;&#x9AD8;&#x7B49;&#x690D;&#x7269;&#x4FDD;&#x80B2;&#x7956;&#x4F20;&#x51FD;&#x6570;&#x3002;NSP1 &#x548C; NSP2 &#x662F;&#x5355;&#x62F7;&#x8D1D;&#x57FA;&#x56E0;&#x5728;&#x8C46;&#x79D1;&#x690D;&#x7269;&#xFF0C;&#x8FD9;&#x610F;&#x5473;&#x7740;&#x8FD9;&#x4E24;&#x79CD;&#x86CB;&#x767D;&#x8D28;&#x5C65;&#x884C;&#x53CC;&#x91CD;&#x76D1;&#x7BA1;&#x804C;&#x80FD;&#x6839;&#x7624;&#x83CC;&#x8BF1;&#x5BFC;&#x4FE1;&#x53F7;&#x4EE5;&#x53CA; SL &#x5408;&#x6210;&#x7684; nonsymbiotic &#x6761;&#x4EF6;&#x4E4B;&#x540E;&#x63A7;&#x5236;&#x4E0B;&#x6E38;&#x7684;&#x76EE;&#x6807;&#x3002;

Strigolactone &#x5408;&#x6210;&#x7684;&#x82DC;&#x84FF;&#x548C;&#x6C34;&#x7A3B;&#x9700;&#x8981;&#x5171;&#x751F;&#x7684;&#x809D;&#x9171;&#x578B;&#x8F6C;&#x5F55;&#x56E0;&#x5B50; NSP1 &#x548C; NSP2&#x3002;

Legume GRAS (GAI, RGA, SCR)-type transcription factors NODULATION SIGNALING PATHWAY1 (NSP1) and NSP2 are essential for rhizobium Nod factor-induced nodulation. Both proteins are considered to be Nod factor response factors regulating gene expression after symbiotic signaling. However, legume NSP1 and NSP2 can be functionally replaced by nonlegume orthologs, including rice (Oryza sativa) NSP1 and NSP2, indicating that both proteins are functionally conserved in higher plants. Here, we show that NSP1 and NSP2 are indispensable for strigolactone (SL) biosynthesis in the legume Medicago truncatula and in rice. Mutant nsp1 plants do not produce SLs, whereas in M. truncatula, NSP2 is essential for conversion of orobanchol into didehydro-orobanchol, which is the main SL produced by this species. The disturbed SL biosynthesis in nsp1 nsp2 mutant backgrounds correlates with reduced expression of DWARF27, a gene essential for SL biosynthesis. Rice and M. truncatula represent distinct phylogenetic lineages that split approximately 150 million years ago. Therefore, we conclude that regulation of SL biosynthesis by NSP1 and NSP2 is an ancestral function conserved in higher plants. NSP1 and NSP2 are single-copy genes in legumes, which implies that both proteins fulfill dual regulatory functions to control downstream targets after rhizobium-induced signaling as well as SL biosynthesis in nonsymbiotic conditions.

Strigolactone biosynthesis in Medicago truncatula and rice requires the symbiotic GRAS-type transcription factors NSP1 and NSP2.

L&eacute;gumineuse GRAS (GAI, RGA, SCR)-tapez PATHWAY1 NODULATION de signalisation des facteurs de transcription (NSP1) et NSP2 sont essentiels pour la nodulation d&#39;induite par le facteur Nod rhizobium. Les deux prot&eacute;ines sont consid&eacute;r&eacute;s comme facteurs de r&eacute;ponse facteur Nod r&eacute;guler l&#39;expression des g&egrave;nes apr&egrave;s signalisation symbiotique. Cependant, l&eacute;gumineuse NSP1 et NSP2 peut &ecirc;tre fonctionnellement remplac&eacute; par nonlegume orthologues, y compris le riz (Oryza sativa) NSP1 et NSP2, ce qui indique que les deux prot&eacute;ines sont fonctionnellement conserv&eacute;s chez les plantes sup&eacute;rieures. Ici, nous montrons que NSP1 et NSP2 sont indispensables pour la biosynth&egrave;se de strigolactone (SL) dans le l&eacute;gumineuses Medicago truncatula et riz. Les plantes mutantes nsp1 ne produisent pas de SLs, tandis que chez le M. truncatula, NSP2 est essentiel pour la conversion des orobanchol en didehydro-orobanchol, qui est la principale SL produite par cette esp&egrave;ce. La biosynth&egrave;se de SL perturb&eacute;e dans nsp1 nsp2 mutant arri&egrave;re-plans est corr&eacute;l&eacute;e avec la r&eacute;duction de l&#39;expression de DWARF27, un g&egrave;ne essentiel pour la biosynth&egrave;se de SL. Riz et M. truncatula repr&eacute;sentent des lign&eacute;es phylog&eacute;n&eacute;tiques distinctes qui divisent environ 150 millions d&#39;ann&eacute;es. C&#39;est pourquoi, nous concluons que la r&eacute;gulation de la biosynth&egrave;se de SL par NSP1 et NSP2 est une fonction ancestrale conserv&eacute;e chez les plantes sup&eacute;rieures. Nsp1 et NSP2 sont des copies uniques des g&egrave;nes chez les l&eacute;gumineuses, qui implique que les deux prot&eacute;ines remplissent deux fonctions r&eacute;glementaires pour contr&ocirc;ler des cibles en aval apr&egrave;s induite par le rhizobium signalisation ainsi que la biosynth&egrave;se de SL dans des conditions non symbiotiques.

Strigolactone biosynth&egrave;se dans Medicago truncatula et riz exige que les facteurs de transcription de GRAS de type symbiotiques NSP1 et NSP2.

Legume GRAS (GAI, RGA, SCR)-geben Sie Transkriptionsfaktoren NODULATION Signalisierung PATHWAY1 (NSP1) und NSP2 sind f&uuml;r Kn&ouml;llchenbakterien Nod Faktor induziert Nodulation. Beide Proteine gelten Nod Faktor Antwort regulierende Genexpression nach symbiotische Signalisierung Faktoren. Jedoch Legume NSP1 und NSP2 k&ouml;nnen funktionell werden ersetzt durch Nonlegume Orthologen, einschlie&szlig;lich Reis (Oryza Sativa) NSP1 und NSP2, die darauf hinweist, dass beide Proteine funktionell in h&ouml;heren Pflanzen konserviert werden. Hier zeigen wir, dass NSP1 und NSP2 f&uuml;r Strigolactone (SL)-Biosynthese in der H&uuml;lsenfrucht Medicago Truncatula und Reis unverzichtbar sind. Mutant nsp1 Pflanzen produzieren keine SLs, M. Truncatula, NSP2 ist notwendig f&uuml;r die Umwandlung von Orobanchol in Didehydro-Orobanchol, welches ist die wichtigste SL produziert von dieser Spezies. Die gest&ouml;rte SL-Biosynthese in nsp1 nsp2 mutant Hintergr&uuml;nde korreliert mit reduzierten Ausdruck des DWARF27, ein Gen f&uuml;r SL-Biosynthese. Reis und M. Truncatula repr&auml;sentieren unterschiedliche phylogenetische Linien, die vor etwa 150 Millionen Jahren aufgeteilt. Daher schlie&szlig;en wir, dass die Regulierung der SL Biosynthese von NSP1 und NSP2 eine uralte Funktion in h&ouml;heren Pflanzen konserviert ist. NSP1 und NSP2 sind Single Copy Gene Leguminosen, was bedeutet dass beide Proteine duale Regulierungsfunktionen nachgelagerte Ziele Steuern nach induziert durch Kn&ouml;llchenbakterien Signalisierung sowie SL Biosynthese in nonsymbiotic Bedingungen erf&uuml;llen.

Strigolactone-Biosynthese in Medicago Truncatula und Reis erfordert die symbiotischen GRAS-Typ-Transkriptionsfaktoren NSP1 und NSP2.

Legume GRAS (GAI, RGA, SCR)-tipo di fattori di trascrizione PATHWAY1 NODULATION segnalazione (NSP1) e NSP2 sono essenziali per rhizobium Nod indotta dal fattore nodulation. Entrambe le proteine sono considerate essere risposta fattore Nod i fattori che regolano l&#39;espressione di gene dopo segnalazione simbiotica. Tuttavia, legume NSP1 e NSP2 pu&ograve; essere funzionalmente sostituito da nonlegume orthologs, tra cui riso (Oryza sativa) NSP1 e NSP2, che indica che entrambe le proteine sono funzionalmente conservate in pi&ugrave; alte piante. Qui, vi mostriamo che NSP1 e NSP2 sono indispensabili per la biosintesi strigolactone (SL) il legume Medicago truncatula e riso. Piante mutanti nsp1 non producono SLs, mentre nella M. truncatula, NSP2 &egrave; essenziale per la conversione di orobanchol dideidro-orobanchol, che &egrave; il principale SL prodotto da questa specie. La biosintesi di SL disturbata in nsp1 nsp2 mutante sfondi correla con ridotta espressione di DWARF27, un gene essenziale per la biosintesi di SL. Riso e M. truncatula rappresentano distinti lignaggi filogenetiche che spaccano circa 150 milioni di anni fa. Pertanto, si conclude che la regolazione della biosintesi di SL da NSP1 e NSP2 &egrave; un&#39;ancestrale funzione conservata in pi&ugrave; alte piante. NSP1 e NSP2 sono geni singola copia nei legumi, che implica che entrambi proteine compiono le funzioni di regolamentazione duale per controllare i bersagli a valle dopo rhizobium-indotta di segnalazione cos&igrave; come la biosintesi SL in condizioni nonsymbiotic.

Biosintesi Strigolactone in riso e Medicago truncatula richiede i fattori di trascrizione GRAS-tipo simbiotici NSP1 e NSP2.

&#x30DE;&#x30E1; &#x30D5;&#x30A9;&#x30A2;&#x30B0;&#x30E9; (&#x30AC;&#x30A4;&#x3001;RGA&#x3001;SCR)-&#x8EE2;&#x5199;&#x56E0;&#x5B50; PATHWAY1 &#x306E;&#x6839;&#x7C92;&#x5F62;&#x6210;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x4F1D;&#x9054;&#x3092;&#x5165;&#x529B; (NSP1) &#x3068; NSP2 &#x304C;&#x6839;&#x7C92;&#x83CC; Nod &#x8A98;&#x5C0E;&#x56E0;&#x5B50;&#x6839;&#x7C92;&#x5F62;&#x6210;&#x306E;&#x305F;&#x3081;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3042;&#x308A;&#x307E;&#x3059;&#x3002;&#x4E21;&#x86CB;&#x767D;&#x8CEA;&#x306F;&#x3046;&#x306A;&#x305A;&#x304F;&#x56E0;&#x5B50;&#x5FDC;&#x7B54;&#x5F8C;&#x5171;&#x751F;&#x30B7;&#x30B0;&#x30CA;&#x30EB;&#x8ABF;&#x7BC0;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x767A;&#x73FE;&#x8981;&#x56E0;&#x3092;&#x3059;&#x308B;&#x3068;&#x898B;&#x306A;&#x3055;&#x308C;&#x307E;&#x3059;&#x3002;&#x305F;&#x3060;&#x3057;&#x3001;&#x30DE;&#x30E1;&#x79D1;&#x690D;&#x7269; NSP1 &#x3068;&#x6A5F;&#x80FD;&#x7684; NSP2 &#x3053;&#x3068; &#xFF08;&#x30A4;&#x30CD;&#xFF09; &#x3092;&#x542B;&#x3080; nonlegume &#x30AA;&#x30EB;&#x30BD;&#x30ED;&#x30B0;&#x306B;&#x3088;&#x3063;&#x3066; NSP1 &#x3068;&#x4E21;&#x86CB;&#x767D;&#x8CEA;&#x304C;&#x9AD8;&#x3044;&#x690D;&#x7269;&#x306E;&#x6A5F;&#x80FD;&#x7684;&#x4FDD;&#x5B58;&#x3055;&#x308C;&#x3066;&#x3044;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3059; NSP2 &#x3092;&#x4EA4;&#x63DB;&#x3057;&#x3066;&#x304F;&#x3060;&#x3055;&#x3044;&#x3002;&#x3053;&#x3053;&#x3067;&#x306F;&#x3001;NSP1 &#x3068; NSP2 &#x304C;&#x30DE;&#x30E1;&#x79D1;&#x690D;&#x7269;&#x30A2;&#x30EB;&#x30D5;&#x30A1;&#x30EB;&#x30D5;&#x30A1; truncatula &#x3068;&#x7C73;&#x30B9;&#x30C8;&#x30EA;&#x30B4;&#x30E9;&#x30AF;&#x30C8;&#x30F3; (SL) &#x306E;&#x751F;&#x5408;&#x6210;&#x304C;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3042;&#x308B;&#x3053;&#x3068;&#x3092;&#x793A;&#x3057;&#x307E;&#x3059;&#x3002;M. truncatula &#x3067; NSP2 &#x3053;&#x306E;&#x7A2E;&#x306B;&#x3088;&#x3063;&#x3066;&#x751F;&#x6210;&#x3055;&#x308C;&#x308B;&#x30E1;&#x30A4;&#x30F3;&#x306E; SL &#x306F;&#x30AA;&#x30ED;&#x30D0;&#x30F3;&#x306E; didehydro &#x30AA;&#x30ED;&#x30D0;&#x30F3;&#x3078;&#x306E;&#x5909;&#x63DB;&#x306E;&#x305F;&#x3081;&#x306B;&#x4E0D;&#x53EF;&#x6B20;&#x3067;&#x3059;&#x306B;&#x5BFE;&#x3057;&#x5909;&#x7570; nsp1 &#x690D;&#x7269; SLs &#x3092;&#x751F;&#x6210;&#x3057;&#x307E;&#x305B;&#x3093;&#x3002;Nsp1 nsp2 &#x5909;&#x7570;&#x80CC;&#x666F;&#x306B;&#x90AA;&#x9B54;&#x306E; SL &#x306E;&#x751F;&#x5408;&#x6210;&#x306E; SL &#x306E;&#x751F;&#x5408;&#x6210;&#x907A;&#x4F1D;&#x5B50; DWARF27 &#x306E;&#x767A;&#x73FE;&#x3068;&#x76F8;&#x95A2;&#x3057;&#x307E;&#x3059;&#x3002;&#x7C73;&#x3068; M. truncatula &#x7D04; 1 &#x5104; 5000 &#x4E07;&#x5E74;&#x524D;&#x306B;&#x5206;&#x5272;&#x500B;&#x5225;&#x306E;&#x7CFB;&#x7D71;&#x306E;&#x7CFB;&#x7D71;&#x3092;&#x8868;&#x3057;&#x307E;&#x3059;&#x3002;&#x3057;&#x305F;&#x304C;&#x3063;&#x3066;&#x3001;&#x6211;&#x3005; NSP1 &#x3068; NSP2 SL &#x751F;&#x5408;&#x6210;&#x306E;&#x8ABF;&#x7BC0;&#x304C;&#x9AD8;&#x3044;&#x690D;&#x7269;&#x306E;&#x4FDD;&#x5B58;&#x3055;&#x308C;&#x3066;&#x3001;&#x5148;&#x7956;&#x4F1D;&#x6765;&#x306E;&#x95A2;&#x6570;&#x3067;&#x3042;&#x308B;&#x3068;&#x7D50;&#x8AD6;&#x3057;&#x307E;&#x3059;&#x3002;NSP1 &#x3068; NSP2 &#x5358;&#x4E00;&#x30B3;&#x30D4;&#x30FC;&#x907A;&#x4F1D;&#x5B50;&#x306E;&#x30DE;&#x30E1;&#x79D1;&#x690D;&#x7269;&#x3001;&#x4E21;&#x86CB;&#x767D;&#x8CEA;&#x6839;&#x7C92;&#x83CC;&#x304C;&#x8A98;&#x5C0E;&#x3055;&#x308C;&#x305F;&#x30B7;&#x30B0;&#x30CA;&#x30EA;&#x30F3;&#x30B0;&#x5F8C; SL &#x751F;&#x5408;&#x6210; nonsymbiotic &#x306E;&#x6761;&#x4EF6;&#x3067;&#x3060;&#x3051;&#x3067;&#x306A;&#x304F;&#x4E0B;&#x6D41;&#x6A19;&#x7684;&#x3092;&#x5236;&#x5FA1;&#x3059;&#x308B;&#x30C7;&#x30E5;&#x30A2;&#x30EB;&#x306E;&#x898F;&#x5236;&#x306E;&#x6A5F;&#x80FD;&#x3092;&#x679C;&#x305F;&#x3059;&#x3053;&#x3068;&#x3092;&#x610F;&#x5473;&#x3067;&#x3059;&#x3002;

&#x30A6;&#x30DE;&#x30B4;&#x30E4;&#x30B7; truncatula &#x304A;&#x3088;&#x3073;&#x30A4;&#x30CD;&#x306B;&#x304A;&#x3051;&#x308B;&#x30B9;&#x30C8;&#x30EA;&#x30B4;&#x30E9;&#x30AF;&#x30C8;&#x30F3;&#x751F;&#x5408;&#x6210;&#x5171;&#x751F;&#x30D5;&#x30A9;&#x30A2;&#x30B0;&#x30E9;&#x578B;&#x8EE2;&#x5199;&#x56E0;&#x5B50; NSP1 &#x3068; NSP2 &#x304C;&#x5FC5;&#x8981;&#x3067;&#x3059;&#x3002;

&#xCF69;&#xACFC; &#xC2DD;&#xBB3C; (&#xC0C1;&#xAC00;, RGA, SCR) &#xADF8; &#xB77C;-&#xC804;&#xC0AC; &#xC778;&#xC790; PATHWAY1 NODULATION &#xC2E0;&#xD638; &#xC785;&#xB825; (NSP1) &#xACE0; NSP2 rhizobium &#xB044; &#xB355; &#xC784; &#xD329;&#xD130; &#xC720;&#xB3C4; &#xB41C; nodulation&#xC5D0; &#xB300; &#xD55C; &#xD544;&#xC218;&#xC801;&#xC778; &#xC694;&#xC18C;&#xC785;&#xB2C8;&#xB2E4;. &#xB450; &#xB2E8;&#xBC31;&#xC9C8;&#xC740; &#xB044; &#xB355; &#xC784; &#xD329;&#xD130; &#xC751;&#xB2F5; &#xC694;&#xC778; &#xACF5;&#xC0DD; &#xC2E0;&#xD638; &#xD6C4; &#xC870;&#xC808; &#xC720;&#xC804;&#xC790; &#xBC1C;&#xD604; &#xB420; &#xC5EC;&#xACA8;&#xC9C4;&#xB2E4;. &#xADF8;&#xB7EC;&#xB098;, &#xCF69;&#xACFC; &#xC2DD;&#xBB3C; NSP1 &#xBC0F; &#xAE30;&#xB2A5;&#xC744; &#xC218;&#xD589; &#xD558;&#xB294; NSP2 &#xC218; NSP1 &#xBC0F; NSP2, &#xB450; &#xB2E8;&#xBC31;&#xC9C8;&#xC5D0;&#xC11C; &#xB354; &#xB192;&#xC740; &#xC2DD;&#xBB3C; &#xBCF4;&#xC874; &#xAE30;&#xB2A5;&#xC801;&#xC73C;&#xB85C; &#xB098;&#xD0C0;&#xB0B4;&#xB294; &#xC300; (Oryza sativa)&#xB97C; &#xD3EC;&#xD568; &#xD558; &#xC5EC; nonlegume orthologs&#xB85C; &#xB300;&#xCCB4;. &#xC5EC;&#xAE30;, &#xC6B0;&#xB9AC;&#xAC00; NSP1 &#xBC0F; NSP2 strigolactone (SL) &#xC0DD; &#xD569;&#xC131; &#xCF69;&#xACFC; &#xC2DD;&#xBB3C; &#xAC1C;&#xC790;&#xB9AC;&#xC18D; truncatula&#xC640; &#xC300;&#xC5D0; &#xB300; &#xD55C; &#xD544;&#xC218; &#xB418;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4; &#xBCF4;&#xC5EC;&#xC90D;&#xB2C8;&#xB2E4;. &#xBC18;&#xBA74; M. truncatula NSP2 didehydro orobanchol orobanchol&#xC758; &#xBCC0;&#xD658;&#xC5D0; &#xB300; &#xD55C; &#xD544;&#xC218;&#xC774; &#xC218; &#xC885;&#xC5D0; &#xC758;&#xD574; &#xC0DD;&#xC0B0; &#xD558;&#xB294; &#xC8FC;&#xC694; SL&#xC740; &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; nsp1 &#xC2DD;&#xBB3C;&#xC5D0;&#xB294; SLs, &#xC0DD;&#xC0B0; &#xD558;&#xC9C0; &#xC54A;&#xC2B5;&#xB2C8;&#xB2E4;. Nsp1 nsp2 &#xB3CC;&#xC5F0;&#xBCC0;&#xC774; &#xBC30;&#xACBD;&#xC5D0;&#xC11C; &#xBC29;&#xD574; SL &#xC0DD; &#xD569;&#xC131; &#xAC10;&#xC18C; &#xC2DD; DWARF27, SL &#xC0DD; &#xD569;&#xC131;&#xC5D0; &#xD544;&#xC218;&#xC801;&#xC778; &#xC720;&#xC804;&#xC790;&#xC758; &#xC5F0;&#xAD00; &#xC2DC;&#xD0B5;&#xB2C8;&#xB2E4;. &#xC300;&#xACFC; M. truncatula &#xC57D; 150 &#xC5B5; &#xB144; &#xC804; &#xBD84;&#xD560; &#xB69C;&#xB837;&#xD55C; phylogenetic &#xACC4;&#xBCF4;&#xB97C; &#xB098;&#xD0C0;&#xB0C5;&#xB2C8;&#xB2E4;. &#xB530;&#xB77C;&#xC11C;, &#xC6B0;&#xB9AC;&#xB294; SL NSP1 &#xBC0F; nsp2&#xC5D0; &#xC758;&#xD574; &#xC0DD; &#xD569;&#xC131;&#xC758; &#xADDC;&#xC81C;&#xB294; &#xB354; &#xB192;&#xC740; &#xC2DD;&#xBB3C;&#xC5D0; &#xBCF4;&#xC874; &#xC870;&#xC0C1;&#xC758; &#xD568;&#xC218; &#xACB0;&#xB860; &#xC9C0;&#xC5C8;&#xB2E4;. NSP1 &#xBC0F; nsp2&#xB294; &#xB2E8;&#xC77C; &#xBCF5;&#xC0AC; &#xC720;&#xC804;&#xC790; &#xCF69;&#xB294; &#xC758;&#xBBF8; &#xD558;&#xB294; &#xB450; &#xB2E8;&#xBC31;&#xC9C8; SL nonsymbiotic &#xC870;&#xAC74;&#xC5D0;&#xC11C; &#xC0DD; &#xD569;&#xC131; &#xBFD0;&#xB9CC; &#xC544;&#xB2C8;&#xB77C; &#xC2E0;&#xD638; rhizobium &#xC720;&#xB3C4; &#xD6C4; &#xB2E4;&#xC6B4;&#xC2A4;&#xD2B8;&#xB9BC; &#xB300;&#xC0C1;&#xC5D0; &#xCEE8;&#xD2B8;&#xB864; &#xB4C0;&#xC5BC; &#xADDC;&#xC815; &#xD558;&#xB294; &#xAE30;&#xB2A5;&#xC744; &#xC218;&#xD589; &#xD569;&#xB2C8;&#xB2E4;.

&#xAC1C;&#xC790;&#xB9AC;&#xC18D; truncatula &#xBC0F; &#xC300; Strigolactone &#xC0DD; &#xD569;&#xC131; &#xACF5;&#xC0DD; &#xADF8; &#xB77C; &#xD615; &#xC804;&#xC0AC; &#xC778;&#xC790; NSP1 &#xBC0F; NSP2 &#xD544;&#xC694;&#xD569;&#xB2C8;&#xB2E4;.

Legumbre GRAS (GAI, RGA, SCR)-tipo de factores de transcripci&oacute;n PATHWAY1 se&ntilde;alizaci&oacute;n de NODULACI&Oacute;N (NSP1) y NSP2 son esenciales para la nodulaci&oacute;n de inducida por el factor de cabeceo de rhizobium. Ambas prote&iacute;nas se consideran factores de Nod factor respuesta reguladora expresi&oacute;n g&eacute;nica despu&eacute;s de se&ntilde;alizaci&oacute;n simbi&oacute;tica. Sin embargo, leguminosas NSP1 y NSP2 puede ser funcionalmente sustituido por ort&oacute;logos de desestabilizaci&oacute;n, incluyendo el arroz (Oryza sativa) NSP1 y NSP2, indicando que ambas prote&iacute;nas funcionalmente se conservan en las plantas superiores. A continuaci&oacute;n, os mostramos que NSP1 y NSP2 son indispensables para la bios&iacute;ntesis de Estrigolactona (SL) en la leguminosa Medicago truncatula y arroz. Nsp1 mutante plantas no producen SLs, mientras que en M. truncatula, NSP2 es esencial para la conversi&oacute;n de orobanchol en didehidro-orobanchol, que es el principal SL producido por esta especie. La bios&iacute;ntesis de SL disturbada en nsp1 nsp2 mutante fondos se correlaciona con la expresi&oacute;n reducida de DWARF27, un gen esencial para la bios&iacute;ntesis de SL. Arroz y M. truncatula representan distintos linajes filogen&eacute;ticos que divide aproximadamente 150 millones de a&ntilde;os. Por lo tanto, concluimos que la regulaci&oacute;n de la bios&iacute;ntesis de SL por NSP1 y NSP2 es una ancestral funci&oacute;n conservada en las plantas superiores. NSP1 y NSP2 son solo-copia genes en las leguminosas, que implica que ambas prote&iacute;nas cumplen dos funciones reguladoras para controlar objetivos aguas abajo despu&eacute;s de rhizobium-inducida de se&ntilde;alizaci&oacute;n as&iacute; como de la bios&iacute;ntesis de SL en condiciones nonsymbiotic.

Bios&iacute;ntesis de Estrigolactona en Medicago truncatula y arroz requiere los factores de transcripci&oacute;n tipo GRAS simbi&oacute;ticos NSP1 y NSP2.

Legumes (Fabaceae or Leguminosae) are unique among cultivated plants for their ability to carry out endosymbiotic nitrogen fixation with rhizobial bacteria, a process that takes place in a specialized structure known as the nodule. Legumes belong to one of the two main groups of eurosids, the Fabidae, which includes most species capable of endosymbiotic nitrogen fixation. Legumes comprise several evolutionary lineages derived from a common ancestor 60 million years ago (Myr ago). Papilionoids are the largest clade, dating nearly to the origin of legumes and containing most cultivated species. Medicago truncatula is a long-established model for the study of legume biology. Here we describe the draft sequence of the M. truncatula euchromatin based on a recently completed BAC assembly supplemented with Illumina shotgun sequence, together capturing ∼94% of all M. truncatula genes. A whole-genome duplication (WGD) approximately 58 Myr ago had a major role in shaping the M. truncatula genome and thereby contributed to the evolution of endosymbiotic nitrogen fixation. Subsequent to the WGD, the M. truncatula genome experienced higher levels of rearrangement than two other sequenced legumes, Glycine max and Lotus japonicus. M. truncatula is a close relative of alfalfa (Medicago sativa), a widely cultivated crop with limited genomics tools and complex autotetraploid genetics. As such, the M. truncatula genome sequence provides significant opportunities to expand alfalfa's genomic toolbox.

The Medicago genome provides insight into the evolution of rhizobial symbioses.

 represents five fast-growing tropical tree species in the Cannabaceae and is the only plant lineage besides legumes that can establish nitrogen-fixing nodules with rhizobium. Comparative analyses between legumes and  allows identification of conserved genetic networks controlling this symbiosis. However, such studies are hampered due to the absence of powerful reverse genetic tools for . Here, we present a fast and efficient protocol for -mediated transformation and CRISPR/Cas9 mutagenesis of . Using this protocol, knockout mutants are obtained within 3 months. Due to efficient micro-propagation, bi-allelic mutants can be studied in the T generation, allowing phenotypic evaluation within 6 months after transformation. We mutated four genes - , and  - that control cytokinin, ethylene, or strigolactone hormonal networks and that in legumes commit essential symbiotic functions. Knockout mutants in  and  displayed developmental phenotypes, namely reduced procambium activity in  and disturbed sex differentiation in  mutants. The symbiotic phenotypes of  and  mutant lines differ from those in legumes. In contrast,  and  are essential for nodule formation, a phenotype similar as reported for legumes. This indicates a conserved role for these GRAS-type transcriptional regulators in rhizobium symbiosis, illustrating the value of  trees as a research model for reverse genetic studies.

CRISPR/Cas9-Mediated Mutagenesis of Four Putative Symbiosis Genes of the Tropical Tree  Reveals Novel Phenotypes.

Nodules harboring nitrogen-fixing rhizobia are a well-known trait of legumes, but nodules also occur in other plant lineages, with rhizobia or the actinomycete  as microsymbiont. It is generally assumed that nodulation evolved independently multiple times. However, molecular-genetic support for this hypothesis is lacking, as the genetic changes underlying nodule evolution remain elusive. We conducted genetic and comparative genomics studies by using  species (Cannabaceae), the only nonlegumes that can establish nitrogen-fixing nodules with rhizobium. Intergeneric crosses between  and its nonnodulating relative  demonstrated that nodule organogenesis, but not intracellular infection, is a dominant genetic trait. Comparative transcriptomics of  and the legume  revealed utilization of at least 290 orthologous symbiosis genes in nodules. Among these are key genes that, in legumes, are essential for nodulation, including  () and  (). Comparative analysis of genomes from three  species and related nonnodulating plant species show evidence of parallel loss in nonnodulating species of putative orthologs of , , and  Parallel loss of these symbiosis genes indicates that these nonnodulating lineages lost the potential to nodulate. Taken together, our results challenge the view that nodulation evolved in parallel and raises the possibility that nodulation originated ∼100 Mya in a common ancestor of all nodulating plant species, but was subsequently lost in many descendant lineages. This will have profound implications for translational approaches aimed at engineering nitrogen-fixing nodules in crop plants.

Marijke Hartog

Laboratory of Molecular Biology,
Department of Plant Sciences,
Laboratory of Molecular Biology, Department of Plant Sciences

Wageningen University & Research

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii

Marijke Hartog

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

Wageningen University & Research

Transforming, Genome Editing and Phenotyping the Nitrogen-fixing Tropical Cannabaceae Tree Parasponia andersonii

Laboratory of Molecular Biology,
Department of Plant Sciences,
Laboratory of Molecular Biology, Department of Plant Sciences