STUDY ON GENETIC FIDELITY IN TISSUE CULTURED RAISED PLANTLETS OF CITRUS LIMON (ASSAM LEMON)
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Date
2023
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Abstract
Citrus lemon, a highly significant species both economically and nutritionally, is extensively cultivated in Assam. The demand for this valuable crop has led to the development of efficient methods for large-scale multiplication to meet the increasing requirements of the market. Micropropagation, an advanced tissue culture technique, offers a promising approach for the rapid production of genetically identical plants. In this study, Citrus lemon was micropropagated to assess its potential for large-scale multiplication, and the genetic fidelity of the micropropagated plantlets was evaluated using Simple Sequence Repeat (SSR), Inter-Simple Sequence Repeat (ISSR), and long Retrotransposon (LTR) markers. SSR markers are highly polymorphic, consisting of short repetitive DNA sequences, while ISSR markers target the regions between microsatellites. LTR markers, on the other hand, focus on retrotransposon dispersed throughout the genome
Healthy and vigorously growing shoot-tip were excised from donor plants and placed on the medium. Within a few weeks, these shoots successfully developed into multiple plantlets through the process of organogenesis. The micropropagated plantlets were then acclimatized to the greenhouse environment, and their growth performance was monitored. To determine the genetic stability and fidelity of the micropropagated plantlets. Fifty plant samples including the mother plant and rest in vitro regenerated plantlets are used for testing the genetic fidelity. Total genomic DNA was extracted from both the mother plants and the micropropagated plantlets using standard protocols, and the SSR, ISSR, and LTR markers were employed to assess genetic variation, if any, between the two sets of samples.
The results of the genetic fidelity analysis demonstrated no detectable variation between the micropropagated Citrus lemon plantlets and their mother plants. All the SSR, ISSR, and LTR markers revealed identical banding patterns, indicating the absence of any somaclonal variation during the micropropagation process. The amplified fragments of the micropropagated plantlets precisely matched those of the donor plants, confirming the maintenance of genetic uniformity in the micropropagation protocol. In conclusion, this study demonstrates that, successful micropropagation of Citrus lemon with retained genetic fidelity is of immense significance for large-scale multiplication and commercial production.