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Research into Stock and Wallflower Protoplast and Electrical Cell Fusion

HUANG Jia-zong1, OKADA Yoshiaki2, FU Jia-rui3 (1. Science and Technology Center of GuangDong Agriculture and Farm, Guangzhou 510507, China; 2. Nara Tabaramoto High Agricultural School, Japan; 3.School of Life Sciences,Sun Yat-sen University, Guangzhou 510275, China)  
Through the electrical cell fusion of the stock and the wallflower, t he optimum conditions of the early-period cultivation of protoplast and the opti mum conditions which lead effectively to cellular fusion. The results were as fo llows: As a condition of early-period cultivation that increases the separati on of seedling protoplast of the stock, by cultivation in a basic culture medium that made up 1/2 MS, a sucrose concentration of 4% and a mannitol concentration of 0.6%, it was the most appropriate. In addition, when adding inorganic nitr ogen to glutamine, it got an excellent result. And when adding plant-growth regu lators BA as 1 mg/L it achieved a high rate of separation. Plus, the effects of th e addition of zeatin (maize factor) was good. Due to pre-treatment of seedlings prior to protoplast separation, the rate of separation achieved was high. The su itable temperature ranged from 24 ℃ to 30 ℃. The conditions which gave rise to a high proportion of single pairs through the medium of electrical cell fusi on were stock protoplast density of 1×105, high electrical field intensity of 1 MHz, the pulse voltage in 1.5 kV/cm and pulse width in 70 s;wallflower prot oplast density of 1×105 units/mL, high electrical field intensity of 1 MHz, 1 00 V/cm the pulse voltage in 2.0 kV/cm and pulse width in 30 μs.
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