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Title:: Study on the Preparation and Cultivation of Soybean Callus Protoplasts

Author(s):: SU Tong¹; YAO Lu-ming¹; ZHANG Xin²; WANG Biao¹; WU Tian-long¹; (1.School of Agricultural and Biology, Shanghai Jiao Tong University, Shanghai 200240, China; 2.College of Life and Environment Science, Shanghai Normal University，Shanghai 200234, China)

Keywords:: Soybean; Callus; Protoplasts; Regeneration; Orthogonal design

PACS:: -

DOI:: 10.11861/j.issn.1000-9841.2018.05.0741

Abstract:: Protoplasts with cell totipotency are ideal materials for exploring genetic transformation and gene function of plants. In order to product and culture effectively protoplast of soybean, the callus induced by immature cotyledons of Jiaoda 05-133 was used as materials preparing for protoplasts in this study. Cellulase onozuka R-10, pectolyase Y-23 and macerozyme R-10 were designed according to the orthogonal array, at the same time, the influence of different enzymatic hydrolysis time on the protoplast yield were explored for optimizing protoplast preparation conditions. Two culture methods with agarose bead and liquid culture were compared to establish an efficient protoplasm regeneration system. The results showed that, the enzyme combination containing cellulase onozuka R-10(2%)， Pectolyase Y-23(0.1%)， macerozyme R-10(1%) was the optimal for protoplasts isolation of Jiaoda 05-133 callus. Under this condition, the appropriate time for Jiaoda 05-133 callus protoplasts isolation was 5 h in enzyme solution and the amounts of protoplasts reaching（3.976±0.86）×10⁶·g^-1 with high quality and vigour were attained. Comparing the two culture methods, the results showed that the agarose bead culture with higher plating rate is more suitable for the cell

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Study on the Preparation and Cultivation of Soybean Callus Protoplasts(PDF)

《大豆科学》[ISSN:1000-9841/CN:23-1227/S]

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