Plant Biotechnology Division, Central Institute of Medicinal and Aromatic Plants, Picnic Spot Road, P.O. CIMAP, Lucknow 226015, India.
Mol Biotechnol. 2012 Nov;52(3):262-8. doi: 10.1007/s12033-011-9491-6.
In vitro grown axillary micro shoots of Glycyrrhiza glabra were encapsulated in alginate beads. Following 6 months of normal storage at 25 ± 2°C the re growth of encapsulated G. glabra micro shoots, reached 98% within 30 days of incubation on MS medium supplemented with 0.1 mg/l IAA. Re growth was characterized by the development of both shoot and root from single encapsulated micro shoot. Healthy plants were established to glass house with 95% survival. The genetic fidelity of plants obtained after conversion of alginate beads was ascertained through 10 RAPD and 13 ISSR primers. Of the 10 RAPD primers tested, 6 of them produced 14 clear and reproducible amplicons with an average of 2.3 bands per primer out of which 28.57% were polymorphic generated by only two primers. Eight ISSR primers produced total 37 bands ranging between 300 and 3,500 bp length. Number of scorable bands for each primer varied from 3 to 8 with an average of 4.6 bands per primer. Cluster analysis from ISSR and RAPD showed that all the tested plants including the mother plant distributed in two major groups with similarity coefficient ranging from 0.91 to 0.96 for RAPD and 0.89 to 0.97 for ISSR.
在体外生长的甘草腋生微芽被包封在藻酸盐珠中。在 25±2°C 的正常条件下储存 6 个月后，在补充了 0.1mg/l IAA 的 MS 培养基上培养 30 天，包封的甘草微芽的再生长率达到了 98%。再生长的特征是从单个包封的微芽中同时发育出茎和根。将植物移栽到温室中，成活率达到了 95%。通过 10 个 RAPD 和 13 个 ISSR 引物来确定转化藻酸盐珠后获得的植物的遗传保真度。在测试的 10 个 RAPD 引物中，有 6 个引物产生了 14 个清晰可重复的扩增子，每个引物平均产生 2.3 个带，其中只有 2 个引物产生了 28.57%的多态性。8 个 ISSR 引物共产生了 37 个条带，长度在 300 到 3500bp 之间。每个引物的可评分条带数从 3 到 8 不等，平均每个引物 4.6 个带。ISSR 和 RAPD 的聚类分析表明，包括母株在内的所有测试植物分为两大组，RAPD 的相似系数为 0.91 到 0.96，ISSR 的相似系数为 0.89 到 0.97。
