DataSheet_1_Exploration of the rhizosphere microbiome of native plant Ceanothus velutinus – an excellent resource of plant growth-promoting bacteria.docx

Continuous demand for an increase in food production due to climate change and a steady rise in world population requires stress-resilient, sustainable agriculture. Overuse of chemical fertilizers and monoculture farming to achieve this goal deteriorated soil health and negatively affected its microbiome. The rhizosphere microbiome of a plant plays a significant role in its growth and development and promotes the plant’s overall health through nutrient uptake/availability, stress tolerance, and biocontrol activity. The Intermountain West (IW) region of the US is rich in native plants recommended for low water use landscaping because of their drought tolerance. The rhizosphere microbiome of these native plants is an excellent resource for plant growth-promoting rhizobacteria (PGPR) to use these microbes as biofertilizers and biostimulants to enhance food production, mitigate environmental stresses and an alternative for chemical fertilizer, and improve soil health. Here, we isolated, purified, identified, and characterized 64 bacterial isolates from a native plant, Ceanothus velutinus, commonly known as snowbrush ceanothus, from the natural habitat and the greenhouse-grown native soil-treated snowbrush ceanothus plants. We also conducted a microbial diversity analysis of the rhizosphere of greenhouse-grown native soil-treated and untreated plants (control). Twenty-seven of the 64 isolates were from the rhizosphere of the native region, and 36 were from the greenhouse-grown native soil-treated plants. These isolates were also tested for plant growth-promoting (PGP) traits such as their ability to produce catalase, siderophore, and indole acetic acid, fix atmospheric nitrogen and solubilize phosphate. Thirteen bacterial isolates tested positive for all five plant growth-promoting abilities and belonged to the genera Pantoea, Pseudomonas, Bacillus, and Ancylobacter. Besides, there are isolates belonging to the genus Streptomyces, Bacillus, Peribacillus, Variovorax, Xenophilus, Brevundimonas, and Priestia, which exhibit at least one of the plant growth-promoting activities. This initial screen provided a list of potential PGPR to test for plant health improvement on model and crop plants. Most of the bacterial isolates in this study have a great potential to become biofertilizers and bio-stimulants.</p

A Rationally Designed Agonist Defines Subfamily IIIA Abscisic Acid Receptors As Critical Targets for Manipulating Transpiration

Increasing drought and diminishing freshwater supplies have stimulated interest in developing small molecules that can be used to control transpiration. Receptors for the plant hormone abscisic acid (ABA) have emerged as key targets for this application, because ABA controls the apertures of stomata, which in turn regulate transpiration. Here, we describe the rational design of cyanabactin, an ABA receptor agonist that preferentially activates <i>Pyrabactin Resistance 1</i> (PYR1) with low nanomolar potency. A 1.63 Å X-ray crystallographic structure of cyanabactin in complex with PYR1 illustrates that cyanabactin’s arylnitrile mimics ABA’s cyclohexenone oxygen and engages the tryptophan lock, a key component required to stabilize activated receptors. Further, its sulfonamide and 4-methylbenzyl substructures mimic ABA’s carboxylate and C6 methyl groups, respectively. Isothermal titration calorimetry measurements show that cyanabactin’s compact structure provides ready access to high ligand efficiency on a relatively simple scaffold. Cyanabactin treatments reduce <i>Arabidopsis</i> whole-plant stomatal conductance and activate multiple ABA responses, demonstrating that its <i>in vitro</i> potency translates to ABA-like activity <i>in vivo</i>. Genetic analyses show that the effects of cyanabactin, and the previously identified agonist quinabactin, can be abolished by the genetic removal of PYR1 and PYL1, which form subclade A within the dimeric subfamily III receptors. Thus, cyanabactin is a potent and selective agonist with a wide spectrum of ABA-like activities that defines subfamily IIIA receptors as key target sites for manipulating transpiration

A Rationally Designed Agonist Defines Subfamily IIIA Abscisic Acid Receptors As Critical Targets for Manipulating Transpiration

Increasing drought and diminishing freshwater supplies have stimulated interest in developing small molecules that can be used to control transpiration. Receptors for the plant hormone abscisic acid (ABA) have emerged as key targets for this application, because ABA controls the apertures of stomata, which in turn regulate transpiration. Here, we describe the rational design of cyanabactin, an ABA receptor agonist that preferentially activates <i>Pyrabactin Resistance 1</i> (PYR1) with low nanomolar potency. A 1.63 Å X-ray crystallographic structure of cyanabactin in complex with PYR1 illustrates that cyanabactin’s arylnitrile mimics ABA’s cyclohexenone oxygen and engages the tryptophan lock, a key component required to stabilize activated receptors. Further, its sulfonamide and 4-methylbenzyl substructures mimic ABA’s carboxylate and C6 methyl groups, respectively. Isothermal titration calorimetry measurements show that cyanabactin’s compact structure provides ready access to high ligand efficiency on a relatively simple scaffold. Cyanabactin treatments reduce <i>Arabidopsis</i> whole-plant stomatal conductance and activate multiple ABA responses, demonstrating that its <i>in vitro</i> potency translates to ABA-like activity <i>in vivo</i>. Genetic analyses show that the effects of cyanabactin, and the previously identified agonist quinabactin, can be abolished by the genetic removal of PYR1 and PYL1, which form subclade A within the dimeric subfamily III receptors. Thus, cyanabactin is a potent and selective agonist with a wide spectrum of ABA-like activities that defines subfamily IIIA receptors as key target sites for manipulating transpiration