Cahya Prihatna, Martin J. Barbetti and Susan J. Barker
Frontiers in Microbiology 8 June 2018
The reduced mycorrhizal colonization (rmc) tomato mutant is unable to form mycorrhiza and is more susceptible to Fusarium wilt compared with its wild-type isogenic line 76R. The rmc mutant has a chromosomal deletion affecting five genes, one of which is similar to CYCLOPS. Loss of this gene is responsible for non-mycorrhizality in rmc but not enhanced Fusarium wilt susceptibility. Here, we describe assessment of a second gene in the rmc deletion, designated Solyc08g075770 that is expressed in roots. Sequence analyses show that Solyc08g075770 encodes a small transmembrane protein with putative phosphorylation and glycosylation sites. It is predicted to be localized in the plasma membrane and may function in transmembrane ion transport and/or as a cell surface receptor. Complementation and knock-out strategies were used to test its function. Some putative CRISPR/Cas-9 knock-out transgenic events exhibited Fusarium wilt susceptibility like rmc and some putative complementation lines were 76R-like, suggesting that the tomato Solyc08g075770 functions in Fusarium wilt tolerance. This is the first study to demonstrate that Solyc08g075770 is the contributor to the Tfw locus, conferring tolerance to Fusarium wilt in 76R which was lost in rmc.
See: https://www.frontiersin.org/articles/10.3389/fmicb.2018.01226/full
FIGURE 3. Amino acid sequence analyses of Solyc08g075770. (A) Molecular phylogenetic analysis by Maximum Likelihood method of Solyc08g075770 and similar sequences in different plant species with 500 bootstrap replications. The tree is drawn to scale, with branch lengths measured in the number of substitutions per site. (B) Solyc08g075770 is predicted to be a protein having three transmembrane domains. Gray boxes are the transmembrane domains, and dashed box is an illustrated cell membrane. (C) Multiple alignment of amino acid sequences of Solyc08g075770 and similar sequences in several plants of the Solanaceae family. Sequences that are shaded gray are the predicted transmembrane domains; sequences that are shaded black are phosphorylation sites whereas sequences with dark gray shading are glycosylation sites (Nicotiana tomentosiformis XM_009622423, N. tabacum XM_016650100, N. attenuata XM_019371074, Capsicum annuum XM_016717203, Solanum tuberosum XM_006364398, S. pennellii XM_015227859). Sequences that are shaded blue are sequences that have significant similarity. The intensity of blue color indicates the degree of conservation or sequence identity: the darker the blue the more conserved the sequences.
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