材料部位相比较，茎尖酶解所得原生质体再生能力最强，较幼叶酶解原生质体，形成细胞壁的时间早3d，第一次分裂时间早2d。
关键词：甘蔗， 原生质体，冻存， 活力， 再生
中图分类号：Q945
文献标识码：A
文章编号：1000-3142（2019）04-0427-10
Abstract：In order to obtain high vitality and regeneration ability of sugarcane protoplast， this experiment wasdone to study the protoplast of frozen storage liquid combination， frozen storage temperature， frozen storage time and recovery temperature. The results were as follows：（1）different cryopreservations，different cryopreservation temperatures anddifferent protoplast sources ofdifferent materials had significantdifferences on the vitality of the protoplast of sugarcane. Compared with the combination of three frozen liquiddeposits， in Combination 2 （70% medium +20% serum +10%dMSO）， the recovery activity was the strongest after 30d， as high as 72%. Cryopreserved recovery within 90d， liquid nitrogen -80 ℃ and -196 ℃ freezer， sugarcane protoplast energydifference was not significant， and the vigor was above 75%. But after 90d of frozen-storage， the protoplastdynamic at -196 ℃ was stronger than -80 ℃ after recovery. Fordifferent materials， the protoplastdynamic of the young leaves frozen stored for 30d was up to 79.2%， and the protoplastdynamic of stem tip frozen for 30d was only 42.7%. （2） There was no significantdifference in the time between the first initiation ofdivision and the formation of cell mass in the treatments withdifferent cryopreservation liquids anddifferent cryopreservation temperatures. After the protoplasts were cultured for about 5-6d， the cell wall was basically complete， and the cells began todivide after 6d， and the cell masses formed after 15d of culture. Fordifferent materials， the strongest regeneration ability was found in the protoplasts from stem tip by enzymatic hydrolysis， which cell wall formation showed 3d earlier， and 2d earlier in the first celldivision than those from the juvenile leaves.
Key words：sugarcane， protoplast， frozen storage， vatality， regeneration
植物体细胞融合育种，是根据细胞的全能性和细胞膜的流动性原理，用酶解法去除细胞壁，先将植物不同种、属，甚至科间的原生质体通过人工方法誘导融合，然后进行离体培养，使其再生杂合体植株的技术。其打破了有性杂交不亲和性的界限，广泛地组合各种基因型，从而形成有性杂交方法所无法获得的新型杂合体植株，而且细胞融合技术避免了分离、提纯、剪切、拼接等基因操作，转移的基因基本不存在基因安全性问题，因为被转移的基因本身存在于甘蔗中，在技术和仪器设备上的要求不像基因工程那样复杂，投资少，有利于广泛开展研究和推广（李志勇，2003），加之体细胞杂交来自双亲的遗传物质并非简单的堆积，而是发生了复杂的遗传重组，这正是改良作物所期望的。植物体细胞融合育种是改造细胞遗传物质的有力手段，有着重大的实践意义。
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