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以红三叶野生品种巴东红三叶的叶片为外植体，在ＭＢ－１至ＭＢ－９的固体培养基上诱导愈伤组织，筛选出最佳培养基是ＭＢ－９培养基；以白三叶栽培品种“路易斯安娜”和“Ｈｕｃａ”的叶片为外植体，接种在ＭＢ－９固体培养基上诱导愈伤组织，愈伤组织形成后均经ＭＢ－９固体培养基上继代３～４次，获得胚性愈伤组织后再转至ＭＢ＋３ｍｇ／Ｌ２，４－Ｄ＋０．７ｍｇ／ＬＢＡ＋０．２ｍｇ／ＬＮＡＡ＋２ｍｇ／Ｌ甘氨酸＋５００ｍｇ／ＬＣＨ＋３％蔗糖的液体培养基中，经强化培养两个月后获得胚性细胞悬浮系。实验结果表明，缩短换液时间（每隔３～４ｄ换液一次）可加快胚性细胞悬浮系的建立。
Using the leaves of red clover, the red clover, as the explants, the callus was induced on the solid medium of MB-1 to MB-9, and the best culture medium was selected as MB-9 medium. The leaves of white clover cultivars “Louisiana” and “Huca” were used as explants and inoculated on MB-9 solid medium to induce callus. After the callus was formed, the callus was subcultured on MB-9 solid medium 3 ~ 4 times. After embryogenic callus was obtained, it was transferred to liquid medium of MB + 3mg / L2, 4-D + 0.7mg / LBA + 0.2mg / L NAA + 2mg / L glycine + 500mg / LCH + 3% sucrose, After obtaining embryogenic cell suspension system. Experimental results show that shortening the fluid exchange time (every 3 ~ 4d liquid change once) can accelerate the establishment of embryogenic cell suspension system.