Agriculture

Author(s): Daniel Glover

Author(s): Luke North, Tucker Howey

Author(s): Cade Smith, Kirsten Sanders

Author(s): Chris Caruso, Cauy Williams

Author(s): Lindy Miller

Author(s): Grace McFarlane

Author(s): Chloe Denham

Author(s): Carson Jocelyn

Author(s): Xander Hulse, Laine Smith

Author(s): Myshela Todd

Author(s): Savanna Greer

Perisse, Iuri Viotti; Fan, Zhiqian; Wang, Zhongde; Harris, Ann; White, Kenneth L.; Polejaeve, Irina A. (Utah State University)
Faculty Advisor: PoleJaeva, Irina (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

Cystic Fibrosis (CF) is a recessive human genetic disease that is caused by mutations in the Cystic Fibrosis Transmembrane Conductance Regulator (CFTR) gene. This gene is responsible to transport Cl- and HCO3- anions in epithelial cells. Previously, we generated CFTR-/- lambs using CRISPR/Cas9 and SCNT techniques. The CFTR-/- lambs display many features similar to human CF disease, including meconium ileus (MI), pancreatic fibrosis, portal fibrosis and biliary hyperplasia, small gallbladder, and absence of vas deferens. In CF patients, MI affects only 15-20% of human babies, whereas it was observed in 100% of newborn CFTR-/- lambs and was the primary cause of death. We here hypothesized that the transgenic expression of the ovine CFTR cDNA under regulation of an intestinal-specific expression promoter would promote the correction of MI in CFTR-/- sheep. In this study, we are constructing three potential vectors with different promoters to be evaluated prior to the generation of transgenic animals. Rat intestinal Fatty Acid Binding Protein (iFABP), rat liver Fatty Acid Binding Protein (LFABP), and Villin1 promoters have already been characterized and successfully used for intestinal-specific expression. After digestion and ligation cloning, the three constructs will be sequenced to confirm the presence of all segments (promoter, cDNA, and vector) in the correct orientation. Subsequently, we plan to evaluate the transient gene expression of the constructs in CaCo-2 cells to ensure they are fully functional. Therefore, we will construct the pcDNA3.1>promoter>CFTR expression vector in order to generate intestine-CFTR transgenic CFTR-/- sheep.

Leatham, Dallin (Weber State University)
Faculty Advisor: Domek, Matthew (Weber State University, Microbiology); Oberg, Craig (Weber State University, Microbiology); Culumber, Michelle (Weber State University, Microbiology); Walker, Edward (Weber State University, Biochemistry)

Gouda cheese is known for its buttery flavor which is produced by the compound 2,3-butanedione or more commonly known as diacetyl. To measure the concentration of diacetyl in cheese a colorimetric test for vicinal diketones is used, known as the Voges and Proskauer (VP) method. This method involves using 5.0% a-naphthol and 0.4% creatine-KOH to develop a pink to red color. In the VP method the concentration of diacetyl is directly related to the absorbance. When this method was performed using known concentrations of Diacetyl in dH2O, a linear relationship was found to have a R2 value of .9571 confirming that the VP method is a viable method of testing for diacetyl concentration. It was proposed that by using a GC-MS to measure these concentrations a simpler test could be performed with similar or improved accuracy. Extracting of the diacetyl from Gouda cheese was done using a stomacher and a mixture of 50:50 acetone:dH2O. Before the concentration of cheese was measured dilutions of diacetyl concentrations were run in a 50:50 acetone:dH2O mixture at 0.1 uL/mL, 1 uL/mL, and 10 uL/mL. The concentrations of diacetyl that gouda cheese commonly contains are from 0.01 uL/mL to 0.1 uL/mL. It was found that the concentration of .1 uL/mL of diacetyl could not be distinguished from the background interference of the instrument. Currently it seems that using a GC-MS to determine diacetyl concentrations in gouda cheese might not be a practical method. Further research and a better understanding of the equipment could improve this method in order to obtain readable results.

Tooley, Sarah (Utah State University)
Faculty Advisor: Powell, Jake (College of Agriculture and Applied Sciences, Landscape Architecture and Environment Planning Department)

The dry, arid climate of Utah makes water a limited and precious resource. However, Utahans are the second largest per capita water users in the nation, with over 60% of that water used on outdoor landscapes. High landscape water use in the face of limited availability is a complex socio-cultural phenomenon reaching back to when pioneers first settled Utah. Settlers were from places with cultural perceptions of landscapes and climates much different then what they found when they arrived. Their desire to create a place that felt like home established a cultural landscape vernacular where Utahan's prefer a landscape more lush and green than we have water to support. With the projected increase in Utah's population, the traditional Utah landscape is quickly becoming unsustainable. As an Extension service I designed 5 landscapes following established water wise design principles which are: planning and design, plant selection and hydro zones along with practical turf areas. The purpose of these designs was to demonstrate that water wise designs can both be water efficient as well as functional, sustainable and aesthetically pleasing. The results of these planning and design efforts are currently being installed into residences in two Utah communities. I will share the design and implementation methods and results of these designs and visitor feedback data from one of the sites. This design research showcases a potential method others in Utah can use to make their landscapes more water wise and begin to redefine the vernacular landscape in Utah and the Intermountain West.

Kaundal, Amita; Hansen, Nathaniel; Ganesh, Jyothsna (Utah State University)
Faculty Advisor: Kaundal, Amita (College of Agriculture and Applied Sciences; Plants, Soils, and Climate Department)

Plants are sessile and bound to their origin so they cannot move to defend themselves against adverse environmental conditions. Soil salinity is one of such environmental stress, which limits the growth and development of plants. Salt stress directly affects crop production. It has been predicted that by 2050, about 50% of arable land will be affected by salinity. About 23% of the worldwide farmland is affected by soil salinity and the crop losses due to salinity are estimated to be tens of billion dollars per year. On top of that, an increase in the world population, the demand for food production also increases. Thus, feeding the growing population under adverse conditions is a challenge. This challenge demands to create more resilient crops to adverse conditions and to feed the growing population. In this study we are investigating General Control Non-repressible 4 (GCN4) in Arabidopsis for salt stress tolerance. GCN4 is an AAA+-ATPase (ATPases associated with diverse cellular activities). AAA+ proteins have diverse functions, such as assembly or disassembly of protein complexes, protein folding or unfolding, protein transport or degradation. It has been shown previously that when AtGCN4 overexpressed in Arabidopsis, it plays a significant role in host-pathogen interaction and control stomatal movement upon pathogen infection by degrading RIN4 - 14-3-3 proteins in PM H+-ATPase complex. Besides this GCN4 when overexpressed exhibit drought tolerance. In this study, we have investigated the previously developed overexpressed and silenced GCN4 Arabidopsis transgenic lines along with wild type plants for salt tolerance at different NaCl concentrations of 90mM, 120mM, and 150mM. So far, the studies indicate that the overexpressor lines were able to tolerate up to 150mm of NaCl as compared to the WT and silenced lines. Further studies are in progress to confirm our hypothesis that GCN4 improves the salt tolerance of transgenic plants. The obtained knowledge will help to investigate the common link in the mechanisms involved in various abiotic and biotic stresses.

Black, Brent; Peterson, Savannah (Utah State University)
Faculty Advisor: Black, Brent (College of Agriculture and Applied Science; Plants, Soils, and Climate Department)

Utah has 1,200 acres of commercial peaches (15th largest peach producing state) making it the second most important fruit crop in the state. However, Utah soils are alkaline, and many trees grown on alkaline soils struggle with iron chlorosis. Peaches and other fruit trees are a combination of the desired variety (scion) grafted onto a rootstock. Some peach rootstocks may be better adapted to Utah's alkaline soils. An orchard comparing 12 peach rootstocks was planted in Utah County in 2007 and evaluated in the 2019 season for survival, tree size and iron chlorosis (leaf chlorophyll content, an indicator of alkaline soil tolerance).The scion for all trees was O'Henry. Most rootstocks had good survival after 13 years (>80%). Empyrean 2 and Julior had the poorest survival (< 50%). The largest trees were on Monegro, Empyrean 1 and Nickels rootstocks, and the smallest were on Lovell, Julior and Krymsk 86. Leaf chlorophyll was lowest (most chlorotic) for Lovell, Julior and Empyrean 1. Selecting the best adapted rootstock is essential for an orchard to be successful and profitable. These results will be useful for Utah growers to select trees best adapted to their conditions.

Waldron, Connor; Moley, Laura; Isom, S. Clay (Utah State University)
Faculty Advisor: Isom, S. Clay (College of Agriculture and Applied Sciences; Animal, Dairy, and Veterinary Sciences Department)

In vitro maturation (IVM) is a process that immature oocytes undergo before in vitro fertilization is performed. During IVM, immature oocytes are extracted through aspiration from follicles and receive the necessary hormones to resume meiosis in culture. Success rate of IVM is significantly lower compared to in vivo maturation of oocytes. Oocytes selected for IVM are extracted from small ovarian follicles (1-3mm), which are 4-6 weeks away from complete maturation and subsequent ovulation. During that period, the DNA of the oocyte is undergoing the process of DNA methylation, the addition of a methyl group to cytosines within a CpG context. DNA methylation is an epigenetic change that causes modification to gene expression where methylated gene promoters turn off gene expression. Proper oocyte gene expression is very important in the development of a healthy embryo. Incomplete methylation of aspirated oocytes may be another factor contributing to the low success rates of IVM. We hypothesize that there is a difference in the amount of methylation between oocytes from small ovarian follicles that are further from natural ovulation and oocytes from large ovarian follicles which are developmentally closer to ovulation, with higher levels of methylation in oocytes from large ovarian follicles. To test our hypothesis, large porcine ovarian follicles (7mm and larger) and small porcine ovarian follicles (1-3mm) will be aspirated for ovaries. The oocytes will be stained the two stains, one to visualize the nucleus of the oocyte indicating the presence of DNA and the other to visualize DNA methylation. Fluorescent images will be taken of the oocytes, and the small and large follicle oocyte groups will be compared for genome wide methylation levels.

Colbert, Jeffrey (Weber State University)
Faculty Advisor: Schramm, Katherina (Weber State University, Botany); Hillbig, Bridgette (Weber State University, Botany); Walker, Edward (Weber State University, Chemistry)

Hemp is a multimillion-dollar industry; however, it is only grown in a few states in the US. In prehistoric time, there was a brackish lake that covered the valley of Utah�s Wasatch Front. Because of this lake and different drainage events, the soil chemistry and composition changed depending on the depth and salts concentration of the lake.
This study tested soils from three different locations along the Wasatch Front for their potential to support healthy growth of hemp plants. Mg, Ca, N, P, K concentrations were tested along with pH in soils from different geological strata. In an outdoor environment hemp was grown in five-gallon containers to determine if those soils will support hemp growth yielding high fiber content and quality. Potting soil was used as a control. The same soil characterization tests were conducted after the plants were harvested to show what amendments are needed for a successful crop in following seasons. Mature plants� inflorescences were tested for tetrahydrocannabinol (THC), and cannabidiol (CBD) levels by Utah Department of Agriculture and Food (UDAF). All plants contained less than 0.3% THC. Fiber quantity was measured using confocal microscopy techniques and compared against the different soil collection locations. Water retting was conducted to extract fiber from stalks. Thin Layer Chromatography (TLC) was conducted confirming UDAF�s findings of THC and CBD.