Genetic fidelity assessment of in vitro-regenerated plants of Albizia julibrissin using SCoT and IRAP fingerprinting

Abstract

A protocol was established for callus induction and plant regeneration of Albizia julibrissin Durazz., a multipurpose tree. Calli were induced on hypocotyl explants excised from 10-to 14-d-old in vitro seedlings cultured on Murashige and Skoog (MS) medium supplemented with α -naphthaleneacetic acid (NAA) alone or in combination with 6-benzylaminopurine (BA) or 6-furfurylaminopurine (kinetin). The highest frequency of organogenic callus (82.2± 3.6%) was obtained on MS medium with 10.8 μM NAA and 4.4 μM BA. Calli were then cultured on MS medium with BA or zeatin, singly or in combination, for shoot regeneration. Calli cultured on MS medium with 13.2 μM BA and 4.6 μM zeatin produced the highest frequency of adventitious shoot regeneration (75.3±6.3%). Maximum rooting of shoots (73.3±5%) was achieved using half-strength MS medium with 4.9 μM indole-3-butyric acid. The genetic fidelity of 12 plants acclimatized to the greenhouse was assessed based on analyses of start codon targeted (SCoT) polymorphism and inter-retrotransposon amplified polymorphism (IRAP). The 14 SCoT and 7 IRAP adapted primers produced 71 and 34 scoreable fragments, of which 33 (46%) and 12 (35%) were polymorphic, respectively. The in vitro-raised plants exhibited 0.129-0.438 genetic distance from the mother plant and 0.000-0.788 distance from one another according to the SCoT and IRAP analyses. Although the culture method described here may not be suitable for clonal propagation of elite genotypes, it can be used for conservation of this plant.

Indirect Regeneration and Assessment of Genetic Fidelity of Acclimated Plantlets by SCoT, ISSR, and RAPD Markers in Rauwolfia tetraphylla L.: An Endangered Medicinal Plant

“…SCoT markers can be used effectively for both genetic diversity and similarity studies [51, 52]. SCoT markers were already reported for assessing the genetic stability of several plant species such as Cleome gynandra [53], Pittosporum eriocarpum [48] , Albizia julibrissin [54], and Simmondsia chinensis [55]. Similarly, ISSR primer based assessment of regenerans was reported in Tylophora indica [56], Simmondsia chinensis [57], Rauwolfia tetraphylla L. [58], Spilanthes calva [59], Cucumis melo L. [60], Cornus alba [61], and Zea mays [62].…”

Development and Deployment of High-Throughput Retrotransposon-Based Markers Reveal Genetic Diversity and Population Structure of Asian Bamboo

“…The inter-retrotransposon amplified polymorphism (IRAP) technique produces amplified fragments that are characteristic of a dominant marker [55,56]. Amplification of the IRAP fragment between two LTR-retrotransposons is done using outward-facing primers which anneal to LTR sequences.…”

Genetic diversity of Bletilla striata assessed by SCoT and IRAP markers

“…They are widespread used on account of their simple operation and high detection efficiency [8]. The different DNA-based molecular markers unfold plant variability directly at genetic levels with reliable data required for the estimation of genetic diversity [2], such as inter-simple sequence repeats (ISSR), inter-retrotransposon amplified polymorphism (IRAP), or start codon targeted (SCoT) polymorphism [13]. In this investigation, SCoT and IRAP were used to assess genetic constancy of Bletilla striata.…”