Conservation, ex vitro direct regeneration, and genetic uniformity assessment of alginate-encapsulated nodal cuttings of Sphagneticola calendulacea (L.) Pruski

• Published in: Acta physiologiae plantarum Vol. 40; no. 3; pp. 1 - 10
• Main Authors: Kundu, Suprabuddha, Salma, Umme, Ali, Md. Nasim, Mandal, Nirmal
• Format: Journal Article
• Language: English
• Published: Berlin/Heidelberg Springer Berlin Heidelberg 01.03.2018; Springer Nature B.V
• Subjects: Acclimatization; Acetic acid; Agriculture; Alginic acid; Beads; Benzyladenine; Biomedical and Life Sciences; Calcium; Calcium alginate; Calcium chloride; Composting; Conservation; Cuttings; Deoxyribonucleic acid; DNA; DNA fingerprinting; Encapsulation; Fingerprinting; Formulations; Indoleacetic acid; Life Sciences; Low temperature; Naphthalene; Nucleotide sequence; Original Article; Plant Anatomy/Development; Plant Biochemistry; Plant Genetics and Genomics; Plant Pathology; Plant Physiology; Plantlets; Polymerase chain reaction; Random amplified polymorphic DNA; Regeneration; Sandy soils; Seeds; Shoots; Sodium; Sodium alginate; Sphagneticola; Sucrose; Sugar; Survival; Worms; Encapsulation; Genetic uniformity; Sodium alginate; Germplasm exchange; Calcium chloride
• ISSN: 0137-5881; 1861-1664
• DOI: 10.1007/s11738-018-2633-8

A well-organized procedure was established for the conservation and distribution of Sphagneticola calendulacea (L.) Pruski [synonym Wedelia chinensis (Osbeck) Merrill] for the first time, using alginate-encapsulated nodal segments (NSs) as synthetic seeds. The ideal beads were obtained through a combination of 2.5% sodium alginate and 75 mM calcium chloride with 84.40 ± 2.20% rate of shoot emergence. The maximum regeneration (88.84 ± 2.24%) from synthetic seeds was achieved on liquid 1/2Murashige and Skoog (MS) medium in comparison to its other formulations. Furthermore, superior frequency (91.09 ± 2.24%) of complete plantlet (having both shoots and roots) formation was achieved when synthetic seeds were cultured on liquid 1/2MS (1.5% sucrose) fortified with 1.0 mg L −1 N 6 -benzyladenine plus 0.25 mg L −1 α-naphthalene acetic acid. Synthetic seeds could be effectively stored at low temperature (8 °C) up to 90 days with a survival rate of 52.38 ± 3.06%, whereas higher temperature (25 °C) did not support regeneration after 75 days of storage. The plantlets were successfully acclimatized to natural conditions with ~ 89% survival frequency. To by-pass the time-consuming in vitro culture step after encapsulation, synthetic seeds were directly regrown into complete plantlets ex vitro on sand, soil, and vermicompost (1:1:1; w/w). Regeneration rate of 42.22 ± 2.22% was attained when NSs were pretreated on 1/2MS medium containing 4.0 mg L −1 indole-3-acetic acid for 24 h in dark, prior to encapsulation. The random amplified polymorphic DNA and intersimple sequence repeat fingerprinting profiles demonstrated genetic uniformity amongst the regenerated plantlets, in vitro mother plant, as well as in vivo wild plant.