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《Heilongjiang Agricultural Sciences》 2017-10
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High Frequency Plant Regeneration from Mesophyll Protoplasts in Chicory(Cichorium endivia L.)

CHEN Xu-qing;LI Hong-chao;DU Yun-peng;ZHANG Ming-fang;XUE Jing;YANG Feng-ping;ZHANG Xiu-hai;Beijing Agro-biotechnology Research Center;  
In order to make attempts on somatic hybridization and gene transformation,aprocedure for high frequency plant regeneration from chicory(Cichorium endivia L.)mesophyll protoplasts was developed by using leaves of an in-vitro seedling culture as starting materials.Protoplasts with yields of 4.5~5.9×106,were routinely released from one gram fresh leaf which was digested in 10 mL of an enzyme solution for 12 h.After careful rinsing and purification,viability of protoplasts ranged from 78% to 85%.With the aim at selecting a suitable culture medium,three different media were employed to compare their efficiency in protoplast culture.The results showed that the medium "modified KMP"proved the best,leading to highest rates in cell division(48.0%)and plating efficiency(27.5%),respectively.It was found the length of culture period affected either embryogenesis or regeneration of protoplasts,optimal period was 28~41 dresulting in highest percentages of both embryogenic callus(81.2%)and regeneration(70.9%),followed by 55.0% and 45.2% obtained from sub-optimal period 42~55 d,while poor rates(22.5% and 16.5%)occurred in prolonged period 56~69 d.A part of plantlets regenerated from protoplasts were able to root on hormone-free 1/2 MS,whereas rests produced roots after transfer upon the rooting medium 1/2 MS +IBA 0.3 mg·L~(-1)+ 0.1% active charcoal.
【Fund】: 北京市农林科学院科研创新能力建设专项资助项目(KJCX20150204)
【CateGory Index】: S636.9
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