BOS1 is a positive regulator of wounding induced cell death and plant susceptibility to Botrytis

Abstract

Programmed cell death (PCD) is required for many aspects of plant biology, including stress responses, immunity, and plant development including root and flower development. Our understanding of PCD regulation is incomplete, especially concerning regulators involved in multiple divergent processes. The botrytis-suscetible1 (bos1) mutant is one of the genotypes most susceptible to Botrytis cinerea (Botrytis) and has revealed the role of BOS1 in cell death propagation during plant responses to wounding. The bos1-1 allele harbours a T-DNA located in the 5'UTR upstream from the start codon that results in elevated BOS1 transcript levels. Here, we resequenced the bos1-1 genome and found a MAS promoter at the ends of the T-DNAs. Expression of the BOS1 gene under control of the MAS promoter conferred the characteristic bos1-1 Botrytis-sensitivity and wounding phenotypes in wildtype plants. We used Crispr-Cas9 to create new bos1 alleles that disrupt exons. These lines lacked the typical bos1-1 wounding and Botrytis phenotypes, but exhibited reduced fertility, as previously observed in other bos1 T-DNA alleles. With multiple overexpression lines of BOS1, we demonstrate that BOS1 is involved in regulation of cell death propagation in a dosage dependent manner. Our data support that bos1-1 is a gain-of-function mutant and that BOS1 acts as a positive regulator of wounding and Botrytis-induced PCD. Taken together these finding suggest that BOS1 function in both fertility and Botrytis response could be unified under cell death control.

Identifying and correcting repeat-calling errors in nanopore sequencing of telomeres

“…1g, Additional file 2: Table S2 and S3) [24]. These eight different organisms are Arabidopsis thaliana [25,26], Caenorhabditis elegans [27], Gallus gallus (chicken), Drosophila melanogaster [28], Mus musculus (mouse) [29,30], Saccharomyces cerevisiae [31], Schizosaccharomyces pombe, and Danio rerio (zebrafish) [32,33] which are all widely studied and have high-quality reference genomes. At the telomeres, these organisms are known to have (TTA GGG ) n telomeric repeats as humans do (chicken, zebrafish, mouse) [34][35][36], (TTT AGG G) n repeats (A. thaliana) [37], (TG 1-3 ) n repeats (S. cerevisiae) [38], TTAC(A)(C)G 2-8 (S. pombe) [39], (TTA GGC ) n repeats (C. elegans) [40], or retrotransposons (D. melanogaster) [41] (Fig.…”

“…A full list of the datasets used in this study is as indicated in Additional file 2: Table S2. Specifically, runs for each of the following organisms were analyzed: A. thaliana (SRR14474199, SRR16149191) [25,26], C. elegans (SRR15993157, SRR16936857) [27], Chicken (SRR15420785, SRR15420786, SRR15420787, SRR15421342 to SRR15421346), D. melanogaster (SRR15107931 to SRR15107934, SRR15107937) [28], Mouse (SRR11606870, SRR14685232, SRR14685224 to SRR14685243) [29,30], S. cerevisiae (ERR6318522, ERR6318523) [31], S. pombe (SRR17382753, SRR18210325), and Zebrafish (SRR17257555, SRR15037325) [32,33].…”

“…Details on the Human Nanopore and PacBio Datasets used for this study are available in the sub-section on Nanopore and PacBio Datasets in the Methods Section [12-15, 17, 18, 42]. The full list of Nanopore and PacBio datasets for other organisms analyzed in this study is available in Additional file 2: Table S2 and were obtained from the corresponding publications [25][26][27][28][29][30][31][32][33], though we would also like to note that some datasets we had obtained from the SRA database do not have a corresponding publication.…”