The Effect of Salinity on Grape Embryo Growth and Physiology By KayLee Weeden and Teresa Giandonato Hello my name is KayLee Weeden and my name is Teresa Giandonato we are from Sheridan college and we have been working for two semesters testing the affect of salinity on Grape embryo growth and physiology.

Introduction All over the world, salinity is affecting crop production. This “ecological restriction” has become one of the largest threats facing major crops. Salt restricts tissue and organ development. Objective: Improve grapevine tolerance to high salt environments in two varieties of grapes. Frontenac (Vitis riparia x Landot) and Thompson Seedless (Vitis vinifera). So the affects of salinity in grape embryos why is that important? And why would it be important in wyoming. Do grapes even grow here. Well right now there are only a handful of grape species that grow in wyoming. Why is that, well if you look outside you’ll see that our surroundings resemble the north pole. Wyoming’s extreme tempetures and extreme salt levels in the soil prohibit growth of grapes in wyoming. Not only is this important to wyoming and grapes but to crops that can’t grow in certain areas of the world.

Methods Preparation: Grape Somatic Embryos (Thompson Seedless, Frontenac) were germinated. Tissue culture media was made in seven treatments of increasing NaCl concentrations. Experiment: All treatments were placed under light in a temperature controlled room for six weeks. Data Collection: Weekly photos were taken of the individual embryos as well as the entire plates. Each plate was rated on a growth scale of 1-4.

Results Our first set of data was taken and the results were that of which we predicted. The embryos exposed to more salinity died compaired to the control embryos.

Thompson Seedless 0.0 mMol 5 mMol 10 mMol 25 mMol 50 mMol 100 mMol Top left is the Control which has no added NaCl from there to the right we have 5 mMol 10 and 25. We saw growth through every plate except the 200 mMol plate. The 200 mMol plate embryos died at week 3 even before then the embryos showed no growth for three weeks. 50 mMol 100 mMol 200 mMol

Results

0.0 mMol 5 mMol 10 mMol 25 mMol Frontenac 50 mMol 100 mMol 200 mMol

Conclusion Salinity has a significant effect on the growth of grape somatic embryos. The embryos that were grown under high salinity conditions showed little to no growth, while the embryos that were grown with minimal salt content showed signs of maturation. This in vitro screening technique will allow us to compare the effects of salinity on grape embryos that have been genetically altered with the SOS2 gene for possible development of salt tolerant grape varieties.

SOS2 Gene An increase in Sodium concentration activates the Salt-overly-sensitive (SOS) pathway. The SOS3/SOS2 complex activates the Na+/H+ exchanger. This removes sodium from the cell. Batelli, G. et al. MOLECULAR AND CELLULAR BIOLOGY, Nov. 2007, p. 7781–7790 Vol. 27, No. 22

Acknowledgements Wyoming INBRE Dr. Sadanand Dhekney, UW Sheridan R&E Center Dr. Ami Erickson, Sheridan College The project was supported by an Institutional Development Award (IDeA) from the National Institute of General Medical Sciences of the National Institutes of Health under Grant # 2P20GM103432. We also appreciate the use of laboratory space, materials and equipment provided by University of Wyoming Sheridan R and E center. Wyoming INBRE

Literature Cited Alam, A., Juraimi, A.S., Rafii, M.Y., Hamid, A. A., (2014). Effect of Salinity on Biomass Yield and Physiological and Stem-Root Anatomical Characteristics of Purslane (Portulaca oleracea L.) Accessions. BioMed Research International: v. 2015 (1-15). Volkmar, K.M., Hu, Y., Steppuhn, H. (1997). Physiological responses of plants to salinity: A review. Canadian Journal of Plant Science: v. 78 (19-27).\ Zhu, M., Shabala, L., Cuin, T.A., Huang, X., Zhou, M., Munns, R., Shabala, S. (2015). Nax Ioci affect SOS1-like Na/H exchanger expression and activity in wheat. Journal of Experimental Botany: v. 67 (835-844).

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