Influence of Water Deficit on Gene Products in Pinus Halepensis

Project Number: 
97R-16
Project Duration: 
24 months
Dates: 
May 1, 1997 to April 30, 1999
Institution of Principle Investigator while on this project: 
Agricultural Research Organization of Israel

Investigators (most current known information)

Associate Professor, The Volcani Center, Agricultural Research Organization of Israel, Institute of Field Crops & Natural Resources, Laboratory of Forestry, PO Box 6, Bet-Dagan, ISRAEL
TEL: +972-3-968-3875 or 968-3678, FAX: +972-3-966-9642, Email: vcgabi@netvision.net.il
Research Scientist, The Volcani Center, Agricultural Research Organization of Israel, Institute of Field Crops & Natural Resources, Laboratory of Forestry, PO Box 6, Bet-Dagan, ISRAEL
Assistant Professor, Department of Forest Science, Texas A&M; University-Kingsville, College Station TX 77843-2135, USA
Professor and Chair, Department of Forest Science, East Carolina University, Howell Science Complex, Greenville NC 27858-4353
TEL: +1-252-328-2418, FAX: +1-252-328-4178, Email: newtonro@mail.ecu.edu
Professor, Department of Agronomy & Horticulture, New Mexico State University, PO Box 30003 / MSC 3Q, Las Cruces NM 88003-8003
TEL: +1-575-646-5113, FAX: +1-575-646-6041, Email: grphilli@nmsu.edu

Proposal Abstract

Two different approaches were used to identify genes or proteins induced by drought stress in Pinus halepensis, allepo pine. One approach used young seedlings to examine proteins induced by water-deficit-stress. Seedlings were subjected to a rapid stress treatment by withholding water or to a slow induction by subjecting them to varying concentrations of glycerol. In roots of seedlings subjected to the rapid stress, total protein content was reduced to 20% that of unstressed within 18 hours. The protein content of hypocotyls decreased at a slower rate. Coomassie staining of total proteins separated by SDS-PAGE did not show differences between stressed and unstressed seedlings. However, SDS-PAGE of hypocotyl boiling-stable proteins did identify bands induced by stress. When biotin was incorporated into proteins produced during treatments, it was possible to detect additional proteins induced by stress. The various bands reached their maximums at different stress levels. An antibody to a peptide found in LP3, a protein induced by drought stress in loblolly pine, was used to examine induction of dehydrins. The antibody was not specific and reacted with several proteins between 15 and 90 kDa. Some of the bands decreased or increased during stress treatments. The second approach used approximately six-month-old seedlings and differential screening of a cDNA library to isolate clones of induced genes. The library was made using roots of stressed seedlings and was screened using probes generated from stressed and unstressed roots. Of the 21,500 clones examined, we identified 156 that represent genes induced by stress. Fifteen of the identified genes were sequenced and northern blot analyses were used to examine the expression of seven of those genes. The genes induced by stress include those encoding a LEA-like protein (late-embryogenesis-abundant), a lignin biosynthetic enzyme (4-coumarate CoA ligase), a cyclophilin, an endochitinase, a sucrose synthase-type protein and some not similar to previously identified genes. Some of the above proteins are similar to those induced by stress in other species. It will be interesting to determine which, if any, of the characterized genes encode the induced proteins identified with our first approach.

Outcome

Articles in Journal

Ffolliott, P. F., G.J. Gottfried, Y. Cohen and G. Schiller. 2003. "Transpiration by dryland oaks: Studies in the south-western United State and Northern Israel." Journal of Arid Environments 55:595-605.

Schiller, G., E.D. Unger, Y. Moshe, S. Cohen and Y. Cohen. 2003. "Estimating water use by scherophyllous species under east Mediterranean climate, II, the transpiration of Quercus calliprinos Webb. in response to silvicultural treatments." Forest Ecology and Management 170:483-495.

Funding

Support for this project came from the USDA Cooperative State Research, Education, and Extension Service