Titie：Isolation of drought stress induced genes and proteins in natalgrass
(Rhynchelytrum repens ) by suppression subtractive hybridization and
two-dimensional electrophoresis
M.S. student：Hua-Hsuan Liang
Advisor：Professor Yih-Ming Chen
Abstract：
Natalgtass, a weed that widely distributed in the world against drought stress
was subjected to physiological and molecular studies. In thephysiological analysis, the
relative water content and water potential weredecreasing after 24 hours PEG
treatment, while chlorophyll and carotenoid were not. In order to identified genes and
proteins regulated by drought stress, suppression subtractive hybridization (SSH) and
two-dimensional electrophoresis (2-DE) were performed. The mixed cDNAs prepared
from natalgrass treated with PEG 6000 (-1 MPa) as testers and cDNAs from
unstressed natalgrass as drivers were hybridized. We found 38 unique expressed
sequence tags (ESTs) in the subtractive cDNA library. After clustering and blasting,
38 ESTs, which predicted to involved in detoxification, TCA cycle, amino acid and
carbohydrate metabolism were considered to responsible for drought tolerance. In the
2-D gel analysis, 10 protein spots were up regulated and 10 protein spots were down
regulated by PEG 6000 treatment.