Singh Reetika, Rai Manoj Kumar, Kumari Nishi
Department of Botany, Mahila Mahavidyalaya, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India,
Appl Biochem Biotechnol. 2015 Sep;177(2):498-510. doi: 10.1007/s12010-015-1758-0. Epub 2015 Jul 25.
A somatic embryogenesis system was developed for Sapindus mukorossi Gaertn. from leaf explants obtained from fresh flushes of a mature tree. Callus was induced from the midrib region of leaf explants on Murashige and Skoog (MS) medium containing different concentrations of 2,4-dichlorophenoxyacetic acid or 6-benzylaminopurine. Callus induction and somatic embryogenesis was significantly influenced by the size, physiological age, and orientation of leaf explants on the culture medium and plant growth regulators. Adaxial-side-up orientation of leaf explants significantly promoted embryogenesis in comparison with abaxial-side-up orientation. Maximum number of somatic embryos was induced on MS medium supplemented with 8.88 μM 6-benzylaminopurine. Scanning electron microscopy of embryogenic callus revealed somatic embryo origin and the development of globular-, heart-, and cotyledonary-stage somatic embryos. The frequency of maturation as well as germination of somatic embryos was higher on MS medium containing 8.88 μM 6-benzylaminopurine than on medium without 6-benzylaminopurine. Plantlets which developed from somatic embryos were acclimatized successfully with 90 % survival.
从一棵成熟树木新长出的叶片外植体中,开发出了无患子的体细胞胚胎发生系统。在含有不同浓度2,4-二氯苯氧乙酸或6-苄基腺嘌呤的Murashige和Skoog(MS)培养基上,从叶片外植体的中脉区域诱导出愈伤组织。愈伤组织的诱导和体细胞胚胎发生受到叶片外植体在培养基上的大小、生理年龄、方向以及植物生长调节剂的显著影响。与背面朝上的方向相比,叶片外植体正面朝上的方向显著促进了胚胎发生。在添加了8.88 μM 6-苄基腺嘌呤的MS培养基上诱导出的体细胞胚数量最多。对胚性愈伤组织的扫描电子显微镜观察揭示了体细胞胚的起源以及球形、心形和子叶期体细胞胚的发育。含有8.88 μM 6-苄基腺嘌呤的MS培养基上体细胞胚的成熟和萌发频率高于不含6-苄基腺嘌呤的培养基。由体细胞胚发育而来的小植株成功驯化,存活率达90%。