Significance: Roots produce hundreds to thousands of small molecules with unknown functions. We targeted the apocarotenoid pathway, which has been linked to numerous developmental processes in Arabidopsis , for a sensitized chemical genetic screen to identify regulators of root development. β-Cyclocitral, a small molecule derived from β-carotene, was identified as a regulator of root stem cell behavior in Arabidopsis as well as in rice and tomato. β-Cyclocitral promotes root stem cell divisions to enhance root growth and branching. In rice, β-cyclocitral enhanced both root and shoot growth during salt stress, which has important implications for agriculture.
The contributions of the Sgs1, Mph1, and Srs2 DNA helicases during mitotic double-strand break (DSB) repair in yeast were investigated using a gap-repair assay. A diverged chromosomal substrate was used as a repair template for the gapped plasmid, allowing mismatch-containing heteroduplex DNA (hDNA) formed during recombination to be monitored. Overall DSB repair efficiencies and the proportions of crossovers (COs) versus noncrossovers (NCOs) were determined in wild-type and helicase-defective strains, allowing the efficiency of CO and NCO production in each background to be calculated. In addition, the products of individual NCO events were sequenced to determine the location of hDNA. Because hDNA position is expected to differ depending on whether a NCO is produced by synthesis-dependent-strand-annealing (SDSA) or through a Holliday junction (HJ)–containing intermediate, its position allows the underlying molecular mechanism to be inferred. Results demonstrate that each helicase reduces the proportion of CO recombinants, but that each does so in a fundamentally different way. Mph1 does not affect the overall efficiency of gap repair, and its loss alters the CO-NCO by promoting SDSA at the expense of HJ–containing intermediates. By contrast, Sgs1 and Srs2 are each required for efficient gap repair, strongly promoting NCO formation and having little effect on CO efficiency. hDNA analyses suggest that all three helicases promote SDSA, and that Sgs1 and Srs2 additionally dismantle HJ–containing intermediates. The hDNA data are consistent with the proposed role of Sgs1 in the dissolution of double HJs, and we propose that Srs2 dismantles nicked HJs. Chromosomal damage that occurs during normal cell division can be repaired using an intact sequence elsewhere in the genome as a template. This process, termed homologous recombination, is crucial for the repair of a particularly deleterious lesion, the DNA double-strand break. Although recombination is a repair process, it can also lead to exchanges of genetic material, generating crossovers (COs) between the involved chromosomes. Repair of the break without exchange of flanking DNA is called a noncrossover (NCO). As COs can uncover recessive mutations or result in large-scale genome rearrangements, understanding how the CO-NCO outcome is regulated is critical to issues of genome stability. The current study examines the distinctive mechanisms whereby three yeast DNA helicases—Mph1, Sgs1, and Srs2—contribute to the repair of a DNA double-strand break.
19 February 2021 in Proceedings of the National Academy of Sciences
Early root growth is critical for plant establishment and survival. We have identified a molecular pathway required for helical root tip movement known as circumnutation. Here, we report a multiscale investigation of the regulation and function of this phenomenon. We identify key cell signaling events comprising interaction of the ethylene, cytokinin, and auxin hormone signaling pathways. We identify the gene Oryza sativa histidine kinase-1 (HK1) as well as the auxin influx carrier gene OsAUX1 as essential regulators of this process in rice. Robophysical modeling and growth challenge experiments indicate circumnutation is critical for seedling establishment in rocky soil, consistent with the long-standing hypothesis that root circumnutation facilitates growth past obstacles. Thus, the integration of robotics, physics, and biology has elucidated the functional importance of root circumnutation and uncovered the molecular mechanisms underlying its regulation. Significance Root growth in soil is complex. With limited sensory input, roots must navigate an environment containing heterogeneities such as rocks and compaction. The ability of roots to penetrate soil is crucial for water and nutrient uptake as well as mechanical anchorage. Here, we describe a molecular framework controlling helical root growth known as circumnutation. Functional characterization of circumnutation is consistent with the hypothesis that it is crucial for exploratory behavior and substrate penetration. This work lays a foundation for an integrated understanding of how roots grow in soil.
During the transition from darkness to light, a suite of light sensors guides gene expression, biochemistry, and morphology to optimize acclimation to the new environment. Ultraviolet, blue, red, and far-red light all have demonstrated roles in modulating light responses, such as changes in gene expression and suppression of stem growth rate. However, green wavebands induce stem growth elongation, a response not likely mediated by known photosensors. In this study, etiolated Arabidopsis (Arabidopsis thaliana) seedlings were treated with a short, dim, single pulse of green light comparable in fluence and duration to that previously shown to excite robust stem elongation. Genome microarrays were then used to monitor coincident changes in gene expression. As anticipated, phytochrome A-regulated, nuclear-encoded transcripts were induced, confirming proper function of the sensitive phytochrome system. In addition, a suite of plastid-encoded transcripts decreased in abundance, including several typically up-regulated after phytochrome and/or cryptochrome activation. Further analyses using RNA gel-blot experiments demonstrated that the response is specific to green light, fluence dependent, and detectable within 30 min. The response obeys reciprocity and persists in the absence of known photosensors. Plastid transcript down-regulation was also observed in tobacco (Nicotiana tabacum) with similar temporal and fluence-response kinetics. Together, the down-regulation of plastid transcripts and increase in stem growth rate represent a mechanism that tempers progression of early commitment to the light environment, helping tailor seedling development during the critical process of establishment.
As part of the Saccharomyces Genome Deletion Project, sets of presumably isogenic haploid and diploid strains that differed only by single gene deletions were constructed. We found that one set of 96 strains (containing deletions of ORFs located between YOR097C and YOR192C) in the collection, which was derived from the haploid BY4741, has an additional mutation in the MSH3 mismatch repair gene.