Highlights d Quantum-dot technology can be used to track trading strategies of mycorrhizal fungi d Increasing exposure to inequality stimulates trade and resource movement d Fungal trade strategies are not uniform across the symbiotic network d Fungi can capitalize on trade by first moving resources to areas of high demand
Parasponia andersonii is a fast-growing tropical tree that belongs to the Cannabis family (Cannabaceae). Together with 4 additional species, it forms the only known non-legume lineage able to establish a nitrogen-fixing nodule symbiosis with rhizobium. Comparative studies between legumes and P. andersonii could provide valuable insight into the genetic networks underlying root nodule formation. To facilitate comparative studies, we recently sequenced the P. andersonii genome and established Agrobacterium tumefaciens-mediated stable transformation and CRISPR/Cas9-based genome editing. Here, we provide a detailed description of the transformation and genome editing procedures developed for P. andersonii. In addition, we describe procedures for the seed germination and characterization of symbiotic phenotypes. Using this protocol, stable transgenic mutant lines can be generated in a period of 2-3 months. Vegetative in vitro propagation of T 0 transgenic lines allows phenotyping experiments to be initiated at 4 months after A. tumefaciens co-cultivation. Therefore, this protocol takes only marginally longer than the transient Agrobacterium rhizogenes-based root transformation method available for P. andersonii, though offers several clear advantages. Together, the procedures described here permit P. andersonii to be used as a research model for studies aimed at understanding symbiotic associations as well as potentially other aspects of the biology of this tropical tree. Video LinkThe video component of this article can be found at https://www.jove.com/video/59971/ 4 . Additionally, it was demonstrated that the root nodule symbiosis in legumes is founded on the much older, and widespread arbuscular mycorrhizal symbiosis 5 . Phylogenomic comparisons suggest that the nitrogen-fixing nodule symbioses of legumes, Parasponia, as well as, the so-called actinorhizal plant species that host diazotrophic Frankia bacteria, have a shared evolutionary origin 6,7,8 . To determine whether the genes identified to be involved in the legume nodule formation are the part of a conserved genetic basis, studies on non-legume species are essential. To this end, we propose to use P. andersonii as a comparative research model, alongside legumes, to identify the core genetic networks underlying root nodule formation and functioning.P. andersonii is a pioneer that can be found on the slopes of volcanic hills. It can meet growth speeds of 45 cm per month and reach lengths of up to 10 meters 9 . P. andersonii trees are wind-pollinated, which is facilitated by the formation of separate male and female flowers 3,10. We recently sequenced and annotated the diploid genome (2n = 20; 560 Mb/1C) of P. andersonii, and assembled draft genome sequences of 2 additional Parasponia species; P. rigida and P. rugosa 6 . This revealed ~35,000 P. andersonii gene models that can be clustered in >20,000 orthogroups together with genes from M. truncatula, soybean (Glycine max), Arabidopsis thaliana, woodland strawberry (Fragaria vesca), Trema orientalis, black cot...
Strains of arbuscular mycorrhizal fungi (AMF) differ markedly in the growth benefits they provide to plants. We investigated whether these differences depend on the chemical form of inorganic phosphorus. The closely related AMF Glomus custos and Rhizophagus irregularis were compared using Plantago lanceolata as the host plant, grown in quartz sand with either soluble orthophosphate or sparingly soluble hydroxyapatite as a sole source of phosphorus. In a growth experiment with AMF-inoculated plants in a climate chamber, sampling at 3-wk intervals enabled a detailed time-resolved analysis of shoot and root phosphorus concentrations and growth performance of P. lanceolata. The ability of AMF to enhance plant growth and deliver phosphate depended strongly on the identity of the available phosphorus source. In orthophosphate-amended substrate only modest differences in plant growth performance (dry matter accumulation and allocation, phosphorus acquisition) were observed between the two AMFs, despite evident AMF root colonization as shown by strain-specific mtLSU qPCR analysis. The treatment with hydroxyapatite however, created stringent growth-limiting conditions and significantly increased the growth benefit provided by R. irregularis over G. custos and the non-mycorrhizal treatment. Plants with R. irregularis could acquire much more phosphorus from apatite compared to G. custos. There were also differences in shoot-to-root dry matter allocation and plant tissue phosphorus concentrations between the R. irregularis and G. custos treatments. Our observations suggest that in experiments on the symbiosis between plants and mycorrhizal fungi more attention should be paid to the chemical form of phosphorus in soil. 2−), may contribute significantly to the total phosphorus content of a soil, up to 75-90%, especially in the
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