红三叶和白三叶愈伤组织的诱导和胚性细胞悬浮系的建立

Induced Calli and Established Embryogenic Cell Suspensions From Trifolium repens and Trifolium pratense

以红三叶（Ｔｒｉｆｏｌｉｕｍｐｒａｔｅｎｓｅ）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（Ｔｒｉｆｏｌｉｕｍｒｅｐｅｎｓｅ）裁培品种“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．愈伤组织形成以后，均在ＭＢ－９固体培养上继代培养３～４次后，获得胚性愈伤组织，然后转至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，２个月后，获得胚性细胞悬浮系．实验结果表明，缩短换液时间，即每隔３～４ｄ换一次，可加快胚性细胞悬浮系的建立．

Using leaves of wild Batong Trifolium pratense as explant, calli were induced on from MB-1 to MB-9 solid medium After screening the best medium was MB-9 Using leaves of cultivated Trifolium repens, namely“Louisiana” and “Huca”,as explant,they were put on the MB-9 solid medium After calli was formed,they were subcultured on the same solid medium for 3～4 times and embryogenic calli were obtained ,then embryogenic calli were transferred into MB liquid medium with 3 mg/L 2,4-D,0 7 mg/L BA,0 2 mg/L NAA,2 mg/L GLY, 500 mg/L CH,3% sucrose After strengthened culturing for 2 months, the embryogenic cell suspension was obtained The test showed that shortening the time of exchanging liquid,namely once 3～4 d, could quicken establishing embryogenic cell suspension