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Lockwood, E.

Publications and source records attributed to Lockwood, E..

2 recordsLinked to original sources

Moss Transplants in the Tundra Reveal Host-Specific Microbiomes and Nitrogen Fixation Responses

In tundra ecosystems, moss-associated microbes are a major source of new nitrogen, yet the relative contributions of environment, host identity, and microbiome composition to variation in nitrogen-fixation rates are difficult to disentangle. To test how environmental change alters moss microbiomes and nitrogen-fixation rates, we used a one-year reciprocal transplant experiment between two Alaskan tundra sites that differ by 5{degrees}C in mean annual temperature. Intact moss cores containing one of three moss species, Hylocomium splendens, Aulacomnium turgidum, and Pleurozium schreberi, were transplanted between sites or returned to their home site. After one year, we quantified nitrogen-fixation rates using 15N incubation and characterized bacterial communities using 16S rRNA gene amplicon sequencing. H. splendens showed consistently low nitrogen-fixation rates with little transplant response, whereas P. schreberi and A. turgidum home and transplant tundra cores generally exhibited higher rates at the cooler, more northern site regardless of origin. In contrast, bacterial community structure changed little following transplantation, with composition driven primarily by moss species. Only in cyanobacteria and some heterotrophic bacterial lineages did we find subtle ASV-level changes. The absence of an association between microbial composition and nitrogen fixation, together with the heterogeneity among moss species, suggests that over short timescales, host physiology and microenvironment play a larger role in the variation of nitrogen-fixation rates than community turnover. The fact that short-term shifts in moss-associated nitrogen-fixation rate are driven primarily by host species identity, rather than microbiome restructuring, has important implications for near-term predictions of nitrogen inputs under Arctic climate change.

microbiology↗

Establishing CRISPR-Cas9 in the sexually dimorphic moss, Ceratodon purpureus.

The development of CRISPR technologies provides a powerful tool for understanding the evolution and functionality of essential biological processes. Here we demonstrate successful CRISPR-Cas9 genome editing in the dioecious moss species, Ceratodon purpureus. Using an existing selection system from the distantly related hermaphroditic moss, Physcomitrium patens, we generated knock-outs of the APT reporter gene by employing CRISPR targeted mutagenesis under expression of native U6 snRNA promoters. Next, we used the native homology-directed repair (HDR) pathway, combined with CRISPR-Cas9, to knock-in two reporter genes under expression of an endogenous RPS5A promoter in a newly developed landing site in C. purpureus. Our results show that the molecular tools developed in P. patens can be extended to other mosses across this ecologically important and developmentally variable group. These findings pave the way for precise and powerful experiments aimed at identifying the genetic basis of key functional variation within the bryophytes and between the bryophytes and other land plants. Significance StatementWe have developed CRISPR-Cas9 genome editing tools and protocols for the sexually dimorphic moss, Ceratodon purpureus to generate gene knock-outs and knock-ins within targeted loci. This work facilitates future functional genomic experiments in this fast growing, haploid, dioecious system and suggests these tools will function across much of moss diversity.

molecular biology↗