Genetic Fidelity of in vitro Propagated Breadfruit (Artocarpus altilis) using Inter Simple Sequence Repeat Markers

A viable and simple method for the large scale production of breadfruit (Artocarpus altilis) using tissue culture is used at the Food and Agricultural Research Extension Institute in line with the Food Security policy of Mauritius. Shoot tip buds of Artocarpus altilis are initiated and multiplied on basal micropropagation medium supplemented with 4.4 mu M Benzyl adenine (BA). Direct systems of regeneration through the culture of organized meristems normally produce true-to-type plants. However, variations among in vitro raised plants have been widely reported. The aim of this study was to use ISSR markers to confirm the clonal fidelity of in vitro derived plantlets of breadfruit. DNA extraction of Artocarpus altilis was performed using a modified CTAB method and then purified using RNase and phenol treatment. The presence of DNA was verified on agarose gel electrophoresis and its yield and quality were determined by spectrophotometry. An average yield of 307.3 mu L DNA was obtained but with some proteins, phenols and polysaccharides contamination was noted. The 63 markers generated by Inter Simple Sequence Repeat (ISSR) markers revealed a mean of 22.2% genetic variation and 77.7% genetic fidelity in the 15 week cultured plantlets and a mean of 1.9% genetic variation and 98.2% genetic fidelity in the 38 weeks cultured plantlets. These findings indicate that shoot meristems can be used as explants for ensuring clonal fidelity of micropropagated A. altilis and confirm the usefulness of ISSR markers as a certification tool for monitoring genetic stability during the regeneration process. (C) 2016 Friends Science Publishers