Changes in cold hardiness were confirmed with relative electrolyte leakage (REL) method in the shoots of two peach cultivars (Prunus persica Janghowon Hwangdo and Odoroki) during cold acclimation and deacclimation. Changes in proline (Pro) content and related gene expressions were also analyzed. Particularly, transcript accumulations of P5CS and P5CR were examined using quantitative real-time RT-PCR. REL in the shoots of two peach cultivars was significantly different during the entire experimental period. Cold hardiness of two cultivars increased gradually to December 2012, and then decreased to April 2013, whereas Pro contents of ‘Janghowon Hwangdo’ and ‘Odoroki’ were reduced from the beginning of the experiment to February 2012 and then increased in the spring. Interestingly, P5CS gene encoding an enzyme, which catalyzes conversion from glutamic acid (Glu) into glutamic-γ-semialdehyde (GSA) in the first step of Pro pathway, showed the contrasting patterns with Pro contents of two cultivars. On the other hand, P5CR gene encoding an enzyme, which catalyzes conversion from Δ1-pyrroline-5-carboxylate(P5C)intoPro in the final step of Pro pathway, showed the similar patterns to Pro contents in two cultivars. Our results demonstrate that Pro responds negatively to low temperatures in the shoot of different peach cultivars, including the supplemental 10 peach cultivars, and expression of both P5CS and P5CR genes could show contrasting patterns from each other. Our results suggest that identification of both P5CS and P5CR genes are required necessarily for accurate analysis of Pro biosynthesis because Pro accumulation is affected more by expression of P5CR gene.

Proline is known as an osmotrotectant to enhance tolerance against both salt and dehydration stresses. A P5CS (δ1 -pyrroline-5-carboxylate synthetase) plays a major role in regulation of synthesis of proline. An overexpression of the mothbean P5CS gene in transgenic tobacco plant increased the levels of proline and osmotolerance. In an attempt to look for the possibility to use content of proline as well as a level of P5CS gene expression as molecular markers for salt tolerance, the amounts of proline and transcript levels of P5CS were measured as functions of either concentration of NaCl or length of treatment period among different species of zoysiagrass. Hybridzoysia showed the highest level of proline (329~mu~textrmg /g.f.w.) among five different species of zoysiagrass at 250 mM NaCl in 24 hours. The level of P5CS transcript was also the highest in the hybridzoysia at 250 mM NaCl in 24 hours. The transcriptions of P5CS gene were induced at the rates of 1.2, 1.2, 1.8, and 1.8, upon treatment of 250 mM NaCl in Z. japonica, Z. matrella, Z. sinica and hybridzoysia respectively. Based on a correlation between the level of P5CS transcript and the proline content among different species of zoysiagrass, a comparative structural analysis of the gene for P5CS from either Z. sinica or hybridzoysia may lead to an understanding of mechanism for salt tolerance shown differently among zoysiagrasses.