Study of Genetic and Phenotypic Variability among Somaclones Induced by BAP and TDZ in Micropropagated Shoot Tips of Banana (Musa spp.) Using RAPD Markers

Abstract

The effects of different concentrations of Benzylaminopurine (BAP) and Thidiazuron (TDZ) on somaclonal variation based on genetic and phenotypic variability among micropropagated shoot tips of banana cultivars; 'Berangan Intan', 'Berangan' and 'Rastali' were studied. With the highest concentrations of both BAP and TDZ up to 44.4µM and 7.5 µM, respectively, most shoots were curled and other morphological changes such as hyperhydricity and undifferentiated shoots were observed. Detectable genetic changes to reveal stable variations among 27 micropropagated variants produced by BAP and TDZ were confirmed using Random Amplified Polymorphic DNA (RAPD) markers. The percentages of polymorphism among somaclones of banana cultivars; 'Berangan Intan', 'Berangan' and 'Rastali' regenerated by TDZ were 24.4, 27.3 and 29.5, respectively which was higher than BAP treatment with the percentages of 16.7, 24.2 and 24.7, respectively. The number of scorable bands among the RAPD profiles obtained with 19 primers tested in this study varied from 6 for opa11 and opk03 to 18 by opc01 in somaclones derived from BAP and 4 for opk03 to 14 with opa04, opb18 and opc04 in the case of TDZ. Cluster analysis of the somaclones regenerated with each of BAP and TDZ resulted in the formation of two major groups, one belonging to Berangan Intan and Berangan (AAA) and the other to Rastali (AAB). From each dendrogram caused by BAP and TDZ, it could be concluded that regenerated somaclones by higher concentrations of both BAP and TDZ accumulated much genetic variation compared to the control and lower concentrations of each of BAP and TDZ.

Cellular, Molecular, and Physiological Aspects of In Vitro Plant Regeneration

“…Several strategies such as morpho-physiological, biochemical, cytological, and DNA-based molecular markers approaches have been employed to maintain the genetic constancy of the in vitro regenerated plantlets. Detection of the genetic homogeneity in micropropagated plants using molecular techniques could be achieved with polymerase chain reaction (PCR)-based techniques such as random amplified polymorphic DNA (RAPD) and inter simple sequence repeat (ISSR) [ 43 , 44 ]. For plant crops that are difficult to breed or have a poor genetic basis, somaclonal variation can be a very useful option for breeders as a new option [ 45 ].…”

“…If the abnormality is the result of DNA modifications, it will be very difficult to reverse. Meanwhile, abnormalities caused by epigenetic modification may be reversible and the resulting abnormal embryos can regenerate to a perfect plantlet [ 43 , 134 , 199 , 200 ].…”

EFFECTIVE CACAO SOMATIC EMBRYO REGENERATION ON KINETIN SUPPLEMENTED DKW MEDIUM AND SOMACLONAL VARIATION ASSESSMENT USING SSRs MARKERS

“…Lopez et al, (2010) used TDZ to induce cacao somatic embryos while in this study used kinetin. According to Bidabadi et al (2010) and Jose et al, (2012), the types and concentrations of PGRs may have caused different level of somaclonal variations. The regenerated cacao plantlets carrying high frequency of allele losses and multiple mutations found in this study support previous finding reported by Lopez et al (2010).…”

The Stimulated Growth of Tissue Cultured Banana and Slipper Orchid as Exposed to Extremely Low Frequency Electromagnetic Field

“…Furthermore, it was also intricate to find out if the plantlets came from the same batch of multiplication; since number of subcultures was also a source of somaclonal variation [30]. Somaclonal variation is a common phenomenon that occurs in the banana tissue culture especially one with the polyploidy cultivar, such as Barangan (AAA) [31]. The somaclonal variation induced in in vitro culture influenced mitotic instability especially in regenerate abnormal metaphase [32].…”