Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum

Bioinformatics tools have been employed for the direct development of gene-based simple sequence repeat (SSR) markers. Through the analysis of 28,056 Mesembryanthemum expressed sequence tag (EST) sequences, a total of 5,851 ESTs containing SSRs were identified, amounting to approximately 17.07 Mb. A...

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Main Authors: Muhanad Akash, Safwan Shiyab, Mohammed Saleh, Shireen M. Hasan, Mahmoud AbuHussein, Wajdy Al-Awaida
Format: Article
Language:English
Published: Hindawi Limited 2023-01-01
Series:Scientifica
Online Access:http://dx.doi.org/10.1155/2023/6624354
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author Muhanad Akash
Safwan Shiyab
Mohammed Saleh
Shireen M. Hasan
Mahmoud AbuHussein
Wajdy Al-Awaida
author_facet Muhanad Akash
Safwan Shiyab
Mohammed Saleh
Shireen M. Hasan
Mahmoud AbuHussein
Wajdy Al-Awaida
author_sort Muhanad Akash
collection DOAJ
description Bioinformatics tools have been employed for the direct development of gene-based simple sequence repeat (SSR) markers. Through the analysis of 28,056 Mesembryanthemum expressed sequence tag (EST) sequences, a total of 5,851 ESTs containing SSRs were identified, amounting to approximately 17.07 Mb. Among these, 938 EST sequences harbored more than one SSR marker, and 788 EST-SSR sequences were found in compound form. The most prevalent types of SSR motifs were mononucleotide repeats (MNRs), accounting for 44%, followed by di-nucleotide repeats (DNRs) at 37%, and trinucleotide repeats (TNRs) at 16%. Notably, TNR or longer SSR motifs primarily consisted of shorter repeat lengths, with only 51 motifs containing 10 or more repeats. The BLASTX analysis successfully assigned functions to 4,623 (79%) of the EST sequences. Among the developed primer sets, 21 primers amplified a total of 65 alleles, with primer PMA79 EST-SSR exhibiting the maximum of six alleles. The polymorphic information content (PIC) values ranged from 0 to 0.76, with a mean of 0.47. The marker index (MI) and discriminating power (D) values reached 0.66 (primer PMA63) and 0.95 (primer PMA20), respectively. Utilizing the unweighted pair group method with arithmetic mean (UPGMA), a dendrogram was constructed, successfully segregating the 24 Mesembryanthemum genotypes into three distinct clusters, with a similarity coefficient ranging from 0.96 to 0.38. In this study, we have developed a total of 83 EST-SSR primer pairs specific to the Mesembryanthemum genus. These newly developed EST-SSRs will serve as valuable tools for researchers, particularly molecular breeders, enabling gene-based identification and trait selection through marker-assisted breeding approaches.
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spelling doaj.art-d1ec2ff2d41f4d66b816f3df76943add2023-11-07T00:00:01ZengHindawi LimitedScientifica2090-908X2023-01-01202310.1155/2023/6624354Development and Validation of Gene-Based SSR Markers in the Genus MesembryanthemumMuhanad Akash0Safwan Shiyab1Mohammed Saleh2Shireen M. Hasan3Mahmoud AbuHussein4Wajdy Al-Awaida5Department of Horticulture and Crop ScienceDepartment of Horticulture and Crop ScienceDepartment of Nutrition and Food TechnologyHamdi Mango Center for Scientific Research (HMCSR)Higher Council for Science and Technology (HCST)Department of Biology and BiotechnologyBioinformatics tools have been employed for the direct development of gene-based simple sequence repeat (SSR) markers. Through the analysis of 28,056 Mesembryanthemum expressed sequence tag (EST) sequences, a total of 5,851 ESTs containing SSRs were identified, amounting to approximately 17.07 Mb. Among these, 938 EST sequences harbored more than one SSR marker, and 788 EST-SSR sequences were found in compound form. The most prevalent types of SSR motifs were mononucleotide repeats (MNRs), accounting for 44%, followed by di-nucleotide repeats (DNRs) at 37%, and trinucleotide repeats (TNRs) at 16%. Notably, TNR or longer SSR motifs primarily consisted of shorter repeat lengths, with only 51 motifs containing 10 or more repeats. The BLASTX analysis successfully assigned functions to 4,623 (79%) of the EST sequences. Among the developed primer sets, 21 primers amplified a total of 65 alleles, with primer PMA79 EST-SSR exhibiting the maximum of six alleles. The polymorphic information content (PIC) values ranged from 0 to 0.76, with a mean of 0.47. The marker index (MI) and discriminating power (D) values reached 0.66 (primer PMA63) and 0.95 (primer PMA20), respectively. Utilizing the unweighted pair group method with arithmetic mean (UPGMA), a dendrogram was constructed, successfully segregating the 24 Mesembryanthemum genotypes into three distinct clusters, with a similarity coefficient ranging from 0.96 to 0.38. In this study, we have developed a total of 83 EST-SSR primer pairs specific to the Mesembryanthemum genus. These newly developed EST-SSRs will serve as valuable tools for researchers, particularly molecular breeders, enabling gene-based identification and trait selection through marker-assisted breeding approaches.http://dx.doi.org/10.1155/2023/6624354
spellingShingle Muhanad Akash
Safwan Shiyab
Mohammed Saleh
Shireen M. Hasan
Mahmoud AbuHussein
Wajdy Al-Awaida
Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
Scientifica
title Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
title_full Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
title_fullStr Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
title_full_unstemmed Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
title_short Development and Validation of Gene-Based SSR Markers in the Genus Mesembryanthemum
title_sort development and validation of gene based ssr markers in the genus mesembryanthemum
url http://dx.doi.org/10.1155/2023/6624354
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