Evolution of specificity at the D14L signaling node for plant development and symbiosis

主题：农业与粮食安全揭示D14 L在基生植物地钱中的功能。KAI2/D14 L在拟南芥和水稻的萌发、幼苗生长以及与丛枝菌根真菌（AM真菌）的有益结合中起着重要作用，但D14 L在基部植物中的功能尚不清楚。另一方面，Karrikins（KARs）是在烟雾中发现的非生物化合物，可刺激追火植物的种子萌发并影响幼苗生长。SL和KAR的受体D14和D14L/KAI2是旁系同源物。SL-不敏感的d14突变体保留被AM真菌定殖的能力。相反，水稻D14L是不可缺少的根的AM真菌的感知和共生相互作用的启动。D14和D14L的共同进化起源解释了SL和KAR信号通路之间的显著相似性，其最终导致对F盒蛋白DWARF3（D3）/更多轴突生长（MAX 2）的共同需求。KAR/KL信号传导组分D14 L、SMAX1和D3的直系同源物存在于所有陆地植物中，而SL信号传导蛋白D14和D53在基础植物中不存在。因此，通过D14-D3-D53的典型SL信号传导是Embryota的创新。尽管如此，SL被发现在基础植物，这确实有外源SL类似物，可能通过D14 L介导的发育反应。D14 L介导的机制可能对早期陆地植物发育和AM共生相互作用很重要。我们将利用我们的集体专业知识来研究D14L在基底植物和火适应物种中的作用。(A)基生苔类包括遗传上易处理的地钱科。我们将CRISPR/Cas9技术应用于菌根物种地钱（Marchantia paleacea），以验证D14 L是植物发育和AM共生所必需的假设。(B)为了评估D14 L在共生中的作用是否在整个植物界中是保守的，将水稻hebibaAOC与M. paleacea和N.将进行衰减的D14L基因。在非菌根拟南芥中引入地钱D14 L未能补充kai2发育表型。如果在水稻中对AM信号进行类似的观察，M.将paleacea D3和SMAX1引入hebibaAOC中并与MpalD14L表达系杂交以验证功能保守性。类似地，将通过水稻D14L的工程等效取代来检查在目的2A中鉴定的赋予KL-配体优先性的D14L变体介导共生信号传导的能力。ENWW：-基因D14L破坏后菌根和地钱的转录组和蛋白质组分析。这些数据集还将与从谷物物种收集的转录组学/蛋白质组学数据进行比较。菌根定殖的评价M。通过生物成像和评估的数量性状的统计分析，

Theme: Agriculture and Food SecurityUnravel the function of D14L in the basal plant Marchantia paleacea. KAI2/D14L plays central roles in germination, seedling growth, and beneficial associations with arbuscular mycorrhizal (AM) fungi in Arabidopsis and rice, but the function of D14L in basal plants remains unknown.Strigolactones (SLs) are plant hormones that regulate plant development and growth of AM fungi. Karrikins (KARs), on the other hand, are abiotic compounds found in smoke that stimulate seed germination of fire-chasing plants and influence seedling growth. The receptors for SLs and KARs, D14 and D14L/KAI2, are paralogs. SL-insensitive d14 mutants retain the ability to be colonized by AM fungi. In contrast, Rice D14L is indispensable for root-perception of AM fungi and for initiation of the symbiotic interaction. The common evolutionary origin of D14 and D14L explains the remarkable parallels between the SL and KAR signaling pathways, which culminate in a shared requirement for the F-box protein DWARF3(D3)/MORE AXILLARY GROWTH (MAX2). Orthologues of the KAR/KL-signaling components D14L, SMAX1 and D3 occur in all land plants, whereas the SL-signaling proteins D14 and D53 are absent from basal plants. Therefore, canonical SL signaling via D14-D3-D53 was an innovation of Embryophyta. Nonetheless SLs are found in basal plants, which indeed have developmental responses to exogenous SL analogs, possibly mediated via D14L. Plausibly, a D14L-mediated mechanism may have been important for early land plant development and AM symbiotic interactions. We will use our collective expertise to investigate the role of D14L in basal land plants and fire-adapted species.(A) The basal liverworts include the genetically tractable Marchantiaceae. We will apply the CRISPR/Cas9 technology to the mycorrhizal species Marchantia paleacea to test the hypothesis that D14L is required for plant development and AM symbiosis in basal plants. (B) To assess whether the role of D14L in symbiosis is conserved across the plant kingdom, genetic complementation of rice hebibaAOC with both M. paleacea and N. attenuata D14L genes will be performed. In non-mycorrhizal Arabidopsis introduction of Marchantia polymorpha D14L failed to complement kai2 developmental phenotypes. If similar observations were made for AM signaling in rice, M. paleacea D3 and SMAX1 will be introduced into hebibaAOC and crossed to the MpalD14L-expressing line to verify functional conservation. Similarly, D14L variants identified in Objective 2A to confer preference for KL-ligands will be examined for their ability to mediate symbiosis signaling by engineering equivalent substitutions of rice D14L.ENWW:- Transcriptome and proteome analysis of Mycorrhiza and Marchantia paleacea upon disruption of the gene D14L. These datasets will additionally be compared to transcriptomics/proteomics data collected from cereal species.- Assessment of mycorrhizal colonization of M. paleacea via biological imaging and statistical analysis of the quantitative traits assessed.