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Publication Type
Conference Proceedings
Author, Analytic
Webster, S. A.; Mitchell, Sylvia A.; Roye, M.; Ahmad, Mohammed H.
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Paper/Section Title
Assessment of genetic stability in repetitive somatic embryogenic cultures of cheese ackee (Blighia sapida) using ISSRs and RAPDs markers
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Proceedings Title
The First International Conference on Tropical Horticulture
Date of Meeting
November 26, 2011.
Place of Meeting
Kingston Jamaica
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Abstract
Inter-simple sequence repeats (ISSRs) and random amplified polymorphic DNA (RAPDs) primers were used as markers to assess the genetic stability of ackee somatic embryogenic cultures, which were subjected to three cycles of repetitive embryogenesis (3G). Recurrent cultures were established from primary somatic embryogenic cultures, which were cultured on MS medium treated with Thidizuron (TDZ) and Abscisic acid (ABA). Secondary somatic embryos were used to establish 3G cultures. The number of somatic embryo and the morphological appearance of the somatic embryoes improved as the number of cycles increased. The genetic stability of the 3G cotyledonary shaped embryos were evaluated using 10 RAPD 10mers (OPB 1 10) and 15 tri-nucleotide ISSR primers. Three of the 10 RAPDs primers and 8 of the 15 ISSR primers generated clear, reproducible and scorable bands. The three RAPDs primers produced a total of 141 bands which were all monomorphic at an average of 5.3 and ranged in size from 950 (OPB 1) 2833 (OPB 7) bp. Eight ISSR primers produced 400 bands at an average 5.5, ranging from 150 1500 bp in size. Two of the 8 ISSR primers produced 1 polymorphic band in one of the samples. In both instances, the band did not represent a gain or loss but rather a change in its position relative to the other homogenous samples. The ISSR primers give a wider coverage of the genome and generated clearer bands that were more reproducible than the RAPD primers. We concluded that the ISSR primers were more robust and more informative as a DNA fingerprinting tool for the assessment of genetic stability of ackee somatic embryogenic cultures. We present herein the details of these results and describe their significance for the use of the somatic embryogenesisprotocol we developed for ackee.....
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