Discipline of Wasteland Research, Central Salt and Marine Chemicals Research Institute, Council of Scientific and Industrial Research, Bhavnagar, Gujarat 364021, India.
Appl Biochem Biotechnol. 2011 Nov;165(5-6):1356-65. doi: 10.1007/s12010-011-9352-6. Epub 2011 Sep 2.
Efficient plantlet regeneration with and without intermediate callus phase was achieved for a selected genotype of Aloe vera L. which is sweet in test and used as a vegetable and source of food. Random amplified polymorphic DNA (RAPD) and inter simple sequence repeats (ISSR) marker assays were employed to evaluate genetic stability of plantlets and validate the most reliable method for true-to-type propagation of sweet aloe, among two regeneration systems developed so far. Despite phenotypic similarities in plantlets produced through both regeneration systems, the differences in genomic constituents of plantlets produced through intermediate callus phase using soft base of inflorescence have been effectively distinguished by RAPD and ISSR markers. No polymorphism was observed in regenerants produced following direct regeneration of axillary buds, whereas 80% and 73.3% of polymorphism were observed in RAPD and ISSR, respectively, in the regenerants produced indirectly from base of the inflorescence axis via an intermediate callus phase. Overall, 86.6% of variations were observed in the plantlets produced via an intermediate callus phase. The occurrence of genetic polymorphism is associated with choice of explants and method used for plantlet regeneration. This confirms that clonal propagation of sweet aloe using axillary shoot buds can be used for commercial exploitation of the selected genotype where a high degree of fidelity is an essential prerequisite. On the other hand, a high degree of variations were observed in plantlets obtained through indirect regeneration and thus cannot be used for the mass multiplication of the genotype; however, it can be used for crop improvement through induction of somaclonal variations and genetic manipulations.
成功建立了高效的不定芽再生体系，其中包括一个中间愈伤组织阶段和一个无中间愈伤组织阶段，用于对选定的基因型的库拉索芦荟（Aloe vera L.）进行遗传稳定性分析。该基因型的库拉索芦荟口感甜美，可作为蔬菜和食品原料。随机扩增多态性 DNA（RAPD）和简单重复间序列（ISSR）标记分析被用来评估植物再生体的遗传稳定性，并验证两种再生系统中最可靠的方法，用于甜芦荟的真实型繁殖。尽管通过两种再生系统产生的植物再生体在表型上相似，但通过软基花序中间愈伤组织阶段产生的植物再生体在基因组组成上的差异可以通过 RAPD 和 ISSR 标记有效区分。在直接从腋芽再生的再生体中没有观察到多态性，而在通过花序轴基部中间愈伤组织阶段间接再生的再生体中，RAPD 和 ISSR 分别观察到 80%和 73.3%的多态性。总体而言，通过中间愈伤组织阶段产生的植物再生体中观察到 86.6%的变异。遗传多态性的发生与外植体的选择和植物再生体的再生方法有关。这证实了使用腋芽对甜芦荟进行克隆繁殖可用于选定基因型的商业开发，其中高度保真度是一个必要的前提。另一方面，通过间接再生获得的植物再生体观察到高度的变异，因此不能用于该基因型的大规模繁殖；然而，它可以用于作物改良，通过诱导体细胞变异和遗传操作。
