TY - BOOK AU - Akhil Sugunan AU - Veena Vigneswaran (Guide) TI - Establishment of genetic fidelity in micropropagated robusta (Musa AAA) plants U1 - 630.28 PY - 2022/// CY - Vellayani PB - Department of Plant Breeding and Genetics, College of Agriculture KW - Plant Breeding and Genetics KW - Robusta banana KW - Musa AAA N1 - M Sc N2 - The present investigation entitled “Establishment of genetic fidelity in micropropagated Robusta (Musa AAA) plants” was carried out at the Department of Plant Breeding and Genetics, Rice Research Station, Vyttila, from September 2020 to October 2021. The study was aimed to assess the genetic fidelity of in vitro propagated banana plants of variety ‘Robusta’ generated by direct organogenesis for ensuring supply of quality planting materials without somaclonal variations. Even though molecular markers are precise and robust method for estimating genetic fidelity, it can only provide information regarding the markers used in the study. So, the work was extended to field level investigation which can be used for validation of molecular analysis. The qualitative and quantitative observations were not restricted to early stages but extended till harvest to make the comparisons to answer farmers concerns, not just to make inferences. One of the qualitative characters studied, ‘bract apex shape’ showed variation in plants derived from subculture S11. It had ‘obtuse’ shaped bract while others had bract shape specified as ‘intermediate’. Other characters compared in the study showed no variation among subcultures. Even though majority of characters were uniform in subcultures from S6 to S11, character modification occurred in S11 for bract apex shape reflects possibility for epi-genetic changes. Significant difference was observed in the subculture S11 from others used in the study for characters - plant height (cm), pseudostem diameter (cm), leaf blade length (cm), leaf blade width (cm), male bud length (cm), male bud circumference (cm), fruit length (cm), number of hands per bunch, number of fingers/hand and bunch weight (kg). All the subcultures showed uniformity for characters - number of leaves, fruit circumference (cm), fruit peel thickness (mm), brix (%) and number of suckers. Out of the 15 characters studied, 10 were showing significant deviation in S11. Two groups were formed based on the sample data from field analysis. Subcultures ranging from S6 to S10 represented one group and S11 was the solo representative of the other group. The mean for all quantitative characters studied was found higher in group which had subcultures from S6 to S10. Subculture S11 was observed as an outlier which joined with the same cluster of other subcultures which 92 are closely related at a higher genetic distance. Similarity for results in ANOVA and D 2 analysis confirmed that the subcultures with non-significant differences in ANOVA were having negligible divergence for quantitative characters and maintained true-totype clonal behaviour in field for the important characters including yield and fruit specifications. Genetic similarity matrix constructed based on simple matching coefficient confirmed molecular level uniformity of subcultures S6 to S10 for the screened markers. The similarity matrix of S11 was clearly found separate from other subcultures. Similarity dendrogram constructed based on UPGMA revealed the separation of S11 cluster away from the clusters of subcultures S6 to S10 placed together. Similarity in results obtained from fields and molecular analysis confirmed a genetic base for variations occurred in S11. The targeted gene specific SSR primers used in the study revealed the polymorphism happened in subculture S11 for markers associated with genes SPL and AP2/EREBP. The error caused in these genes can result delay in phase transition, abnormalities in fruit characters and can induce higher susceptibility to abiotic stress. Similar responses observed in field was in close agreement with the trait-based polymorphism detected in molecular level. Even though true-to-type clones are produced up to ten subcultures, considering the random occurrence of mutation and verification of somaclonal variations associated with higher level of subcultures, the investigation suggests that the subculturing can be safely carried up to subculture 9 for ‘Robusta’ plantlets produced by direct organogenesis. Potential for S10 subculture can be further clarified with use of more molecular markers ER -