A Demographic Profile of Independently Incorporated Native American Foundations and Selected Funds in the United States

This report gives basic demographic information on 60 grantmaking entities grouped into three categories: 1) Native foundations that are independently incorporated; 2) 501c3 Native organizations; and 3) tribal funds. These categories capture the variety of Native controlled approaches currently at work in the field

<i>GmCYP82A3</i>, a Soybean Cytochrome P450 Family Gene Involved in the Jasmonic Acid and Ethylene Signaling Pathway, Enhances Plant Resistance to Biotic and Abiotic Stresses

<div><p>The cytochrome P450 monooxygenases (P450s) represent a large and important enzyme superfamily in plants. They catalyze numerous monooxygenation/hydroxylation reactions in biochemical pathways, P450s are involved in a variety of metabolic pathways and participate in the homeostasis of phytohormones. The <i>CYP82</i> family genes specifically reside in dicots and are usually induced by distinct environmental stresses. However, their functions are largely unknown, especially in soybean (<i>Glycine max</i> L.). Here, we report the function of <i>GmCYP82A3</i>, a gene from soybean <i>CYP82</i> family. Its expression was induced by <i>Phytophthora sojae</i> infection, salinity and drought stresses, and treatment with methyl jasmonate (MeJA) or ethephon (ETH). Its expression levels were consistently high in resistant cultivars. Transgenic <i>Nicotiana benthamiana</i> plants overexpressing <i>GmCYP82A3</i> exhibited strong resistance to <i>Botrytis cinerea</i> and <i>Phytophthora parasitica</i>, and enhanced tolerance to salinity and drought stresses. Furthermore, transgenic plants were less sensitive to jasmonic acid (JA), and the enhanced resistance was accompanied with increased expression of the JA/ET signaling pathway-related genes.</p></div

Enhanced resistance to <i>B</i>. <i>cinerea</i> of transgenic <i>N</i>. <i>benthamiana</i> lines.

<p><b>(A)</b> Phenotypes of the <i>N</i>. <i>benthamiana</i> leaves from WT, EV and overexpression lines (2–3 and 4–1) inoculated with <i>B</i>. <i>cinerea</i>. The mycelia growing on PDA medium was used to infect the transgenic and wild type leaves. Photographs were taken 4 dpi. Bar = 5 mm. <b>(B)</b> Lesion area of inoculated leaves. Lesion diameters were measured at 4 dpi and then the lesion area was calculated. Similar results were observed at least three duplications. SD represented with the bars (Dunnett-t test: * <i>P</i><0.05).</p

The expression profiles of <i>GmCYP82A3</i> response to various abiotic stress and phytohormones.

<p>Three-week-old seedlings were treated with the indicated compounds, and then the samples were collected at 0, 6, 12 and 24 hpt. The relative expression level was normalized to soybean <i>GmTUA</i>. Mean and standard deviation (SD) were calculated from three independent biological replicates. The asterisk at the top of the columns indicate significant differences (Dunnett-t test, * <i>P</i><0.05, ** <i>P</i><0.01).</p

Expression profiles of defense marker genes in transgenic <i>N</i>. <i>benthamiana</i> plants.

<p>Total RNA was extracted from detached leaves of WT, EV and <i>GmCYP82A3</i> overexpression plants at 0, 12, 24 hpi by <i>P</i>. <i>parasitica</i> zoospores. Expression levels were determined by qRT-PCR using gene specific primers and normalized to <i>NbEF1a</i> with three replicate experiments. Data are the means of three replications, error bars indicate SD. The significant differences between WT and transgenic plants are indicated by asterisk (Dunnett-t test, * <i>P</i><0.05, ** <i>P</i><0.01).</p

The effect of <i>GmCYP82A3</i> on root growth under JA treatment.

<p><b>(A)</b> The phenotypes of 2-week-old seedlings of indicated genotypes grown on MS medium without (Control) or with 10 μΜ JA. <b>(B)</b> Root length of 2-week-old WT, EV and <i>GmCYP82A3</i> overexpression seedlings grown on MS medium containing 10 μΜ JA. The experiments were repeated three times with similar results. SD represented with the bars. The asterisk at the top of the columns indicate significant differences (Dunnett-t test, ** <i>P</i><0.01).</p

Seed germination rates of transgenic <i>N</i>. <i>benthamiana</i> under abiotic stress.

<p>The seeds as indicated were cultured on MS medium (<b>A</b>) or containing NaCl (<b>B</b>) or PEG6000 (<b>C</b>). The germination rates were calculated from the percentage of seeds with radicle protruded through the seed coat after cultivation every day. Mean values and SD were obtained from five independent experiments (n = 40). The significant differences between WT and transgenic plants are indicated by asterisk (Dunnett-t test, * <i>P</i><0.05, ** <i>P</i><0.01).</p

The expression pattern of <i>GmCYP82A3</i> during <i>P</i>. <i>sojae</i> infection.

<p><b>(A)</b> The microarray data of relative expression levels of <i>GmCYP82A3</i>. The expression levels in four soybean cultivars with different resistance level (Conrad and General with highly partial resistance, Williams with moderately partial resistance and Sloan is highly susceptible). The relative expression levels were normalized to the mock samples. <b>(B)</b> Expression pattern of <i>GmCYP82A3</i> induced by <i>P</i>. <i>sojae</i> determined by RT-PCR. Soybean leaves of Sloan, Williams and Conrad were inoculated with <i>P</i>. <i>sojae</i> and samples were taken at 0, 3, 6, 12 and 24 hpi. The <i>GmTUA</i> gene serves as a control.</p

Overexpression of GmHsp90s, a Heat Shock Protein 90 (Hsp90) Gene Family Cloning from Soybean, Decrease Damage of Abiotic Stresses in <i>Arabidopsis thaliana</i>

<div><p>Hsp90 is one of the most conserved and abundant molecular chaperones and is an essential component of the protective stress response; however, its roles in abiotic stress responses in soybean (<i>Glycine max</i>) remain obscure. Here, 12 <i>GmHsp90</i> genes from soybean were identified and found to be expressed and to function differentially under abiotic stresses. The 12 GmHsp90 genes were isolated and named <i>GmHsp90A1–GmHsp90A6</i>, <i>GmHsp90B1</i>, <i>GmHsp90B2</i>, <i>GmHsp90C1.1</i>, <i>GmHsp90C1</i>.<i>2</i>, <i>GmHsp90C2</i>.<i>1</i> and <i>GmHsp90C2</i>.<i>2</i> based on their characteristics and high homology to other Hsp90s according to a new nomenclature system. Quantitative real-time PCR expression data revealed that all the genes exhibited higher transcript levels in leaves and could be strongly induced under heat, osmotic and salt stress but not cold stress. Overexpression of five typical genes (<i>GmHsp90A2</i>, <i>GmHsp90A4</i>, <i>GmHsp90B1</i>, <i>GmHsp90C1.1</i> and <i>GmHsp90C2</i>.<i>1</i>) in <i>Arabidopsis thaliana</i> provided useful evidences that GmHsp90 genes can decrease damage of abiotic stresses. In addition, an abnormal accumulation of proline was detected in some transgenic Arabidopsis plants suggested overexpressing GmHsp90s may affect the synthesis and response system of proline. Our work represents a systematic determination of soybean genes encoding Hsp90s, and provides useful evidence that GmHsp90 genes function differently in response to abiotic stresses and may affect the synthesis and response system of proline.</p></div

Fresh weight and pod setting percentage of transgenic Arabidopsis plant under normal or abiotic stresses.

<p>Three-week-old seedlings were saturated with water (control), PEG (8%) or NaCl (150 mM) respectively for 3 d and recovered for 5 d. Fresh weights were measured after 3 d of treatment. For heat stress, three-week-old seedlings were moved to 30°C until pod setting. Shoot fresh weights were measured when pod setting was calculated. (a) Shoot fresh weight of Arabidopsis under heat stress. (b) Pod setting percentage of Arabidopsis under heat stress. (c) Fresh weights of Arabidopsis plants under normal condition, salt and osmotic stress. Error bars indicate SD; n = 20, and plants were prepared from at least five independent plants for each repeat. The means with ‘**’ represent significant differences from each other (<i>P</i><0.01). Control, vector control plants; A2, A4, B1, C1.1, C2.1 represent the <i>GmHsp90A2</i>, <i>GmHsp90A4</i>, <i>GmHsp90B1</i>, <i>GmHsp90C1</i>.1 and <i>GmHsp90C2</i>.1 transgenic lines, respectively.</p