conference schedule and program with abstracts - Horticulture ...

P-28
In vitro selection of HYP tolerant cell lines from “Chardonnay” embryogenic suspension
culture
R. He* 1 , Y. Zhang 1,2 , C. Wang 1 , J. Wu 1,3 , L. Zhu 1 , X. Xu 2 , M.A. Walker 4 , J. Lu 2
1 College of Food Science and Nutritional Engineering, China Agricultural University, Beijing,
China; 2 Center for Viticulture and Small Fruit Research, Florida A&M University, Tallahassee,
Florida, USA; 3 Department of Horticultural Sciences, University of Florida, Gainesville, Florida,
USA; 4 Department of Viticulture and Enology, University of California – Davis, USA
*Corresponding author: rongronghe.smile@gmail.com
Breeding for cold and salt tolerance of woody plants such as grapevine by classical methods is
time consuming. An alternative approach to improve these traits of grapevine is to induce and
select somaclonal variation using in vitro culture strategy. Suspension cultures of “Chardonnay”
Proembryogenic Mass (PEM) were radiated with 10, 40, 60, 100, 500 Gy of Co60 and then
transferred to the selection medium (Murashige and Skoog medium plus 0.5 µM 2, 4-
Dichlorophenoxyacetic acid and 10 mM hydroxyproline). The hydroxyproline resistant cell lines
were obtained from each dose of Co60 treatment (10, 40, 60, 100 Gy) except the 500 Gy, but
most of them became brown after subcultures for several times. However, a stable cell line
derived from radiation of Co60 at 100Gy dose was obtained, which was subsequently confirmed
by hydroxyproline resistant test and proline content. After 4-week exposure to different
concentrations of hydroxyproline, the resistant cell line eventually grew vividly on the MS
medium plus 0.5 µM 2, 4-D and 4.0 mM hydroxyproline, while the control line turned brown.
The resistant PEM also showed a higher free proline content than the control one after treated
with hydroxyproline. The “resistant” PEM line of “Chardonnay” are germinated, and the
plantlets are being used for further evaluation for salt, drought and cold tolerance.
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