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以“Ｐａｇｅ”橘柚（ＣｉｔｒｕｓｒｅｔｉｃｕｌａｔａＢｌａｎｃｏ×Ｃ．ｐａｒａｄｉｓｉＭａｃｆ）胚性悬浮系原生质体和粗柠檬（Ｃ．ｊａｍｂｈｉｒｉＬｕｓｈ）叶肉原生质体为融合试材，摸索了适合柑橘的电融合参数。研究表明，在ＡＣ（交变电场）１２５Ｖ／ｃｍ、ＤＣ（直流脉冲）１２５０Ｖ／ｃｍ、ＡＣ作用时间６０ｓ、ＤＣ脉冲时间５０μｓ、ＤＣ脉冲个数为３、ＤＣ脉冲间隔为０．５ｓ时融合率较高，双核异核体率可以达到１５％以上。利用不同类型原生质体比重的不同，融合后通过控制离心时间，可以收集到更高比例的双核异核体。融合产物经培养，３～４个月再生小植株。染色体计数结合形态学观察表明，再生的１５０株小植株中７８％为四倍体，其余为二倍体叶肉亲本再生类型。再生植株经过氧化物酶同工酶及ＲＡＰＤ分析，表明得到了体细胞杂种，还证明获得了１株叶肉亲本同源四倍体再生类型。
The electrofusion parameters of Citrus reticulata Blanco × C.paradisiMacf embryo suspension protoplasts and C. japumbhiri Lush mesophyll protoplasts were used as fusion materials. The results showed that the fusion rate of AC (alternating current) 125V / cm, DC (DC pulse) 1250V / cm, AC time 60s, DC pulse time 50μs, DC pulse number 3 and DC pulse interval 0.5s Higher, dual nuclear heterozygote rate can reach more than 15%. Taking advantage of different specific protoplasts’ different weights, a higher proportion of binucleated heterokaryons can be collected by controlling centrifugation time after fusion. Fusion products were cultured, 3 to 4 months regenerated plantlets. Chromosome counting combined with morphological observation showed that 78% of the 150 regenerated plantlets were tetraploids, and the rest were diploid parental parental regeneration types. The regenerated plants were analyzed by oxidase isozymes and RAPD, which showed that somatic hybrids were obtained, and one type of autotetraploid regenerated by leaf mesophyll parents was also obtained.