Molecular and morphological characterization of EMS-induced chili pepper mutants resistant to Chili veinal mottle virus
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Abstract. Manzila I, Priyatno TP, Nugroho K, Terryana RT, Lestari P, Hidayat SH. 2020. Molecular and morphological characterization of EMS-induced chili pepper mutants resistant to Chili veinal mottle virus. Biodiversitas 21: 1448-1457. Chili veinal mottle virus (ChiVMV) is a major prevalent virus that potentially reduces the yield of chili pepper by more than 50% in Indonesia. Therefore, the development of a ChiVMV-resistant cultivar is necessary to ensure satisfactory chili pepper production. Mutation breeding is a promising tool available to produce virus-resistant plants via the induction of new alleles due to point mutations within the existing chili germplasm. This research was conducted to assess the genetic variation of ten promising (M6) EMS-induced mutant lines of chili pepper (Capsicum annuum L.) potentially resistant to ChiVMV by their morphological characteristics and SSR profile, as well as identify potential mutants based on agronomic traits. The ten chili pepper mutant lines used in the present study were derived from the shoot tips of a Gelora cultivar treated with 0.5% ethyl methanesulfonate (EMS) as a chemical mutagen. The performance of the respective promising chili pepper mutant lines both at a molecular and morphological level was observed. Phenotypic analysis showed that all the promising chili pepper mutant lines underwent significant morphological changes. Of these, the phenotype coefficient variation (PCV) values were higher than the genotype coefficient variation (GCV) values for all the traits observed. High heritability (h2) was found for the canopy area, length of the fruit stalk, fruit weight, fruit weight per plant, thick fruit flesh, leaf width, leaf length, and fruit storage duration. A total of 52 alleles were detected from the five polymorphic SSR loci with 5.2 average alleles/loci. A dendrogram was generated and revealed two groups with five mutants clustered separately from their parents which could be valuable for further improvement.
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References
REFERENCES
Ajjapplavara PS, Patil SS, Hosmani RM, Patil AA, Gangaprasad S. 2005. Correlation and Path Coefficient Analysis in Chilli. Karnataka J.Agric.Sci.
Alonso, J. M. Stepanova AN, Leisse TJ, Kim CJ, Chen HP. 2003. Genome-wide Insertional Mutagenesis of Arabidopsis thaliana. Science. 301(5633):653–657. doi: 10.1126/science.1086391.
Alvarez A, Fuentes JL, Puldón V, Gómez PJ, Mora L, Duque MC, Gallego G, Tohme JM. 2007. Genetic Diversity Analysis of Cuban Traditional Rice (Oryza sativa L.) Varieties Based on Microsatellite Markers. Genetics and Molecular Biology. 30(4): 1109–1117. doi: 10.1590/S1415-47572007000600014.
Anh TTT, Tran DK, Tran DD, Tran DX. 2018. Identification of Phenotypic Variation and Genetic Diversity in Rice (Oryza sativa L.) Mutants. Agriculture (Switzerland). doi: 10.3390/agriculture8020030.
Anupam A, Imam J, Quatadah SM, Siddaiah A, Das SP, Variar M, Mandal NP. 2017. Genetic Diversity Analysis of Rice Germplasm in Tripura State of Northeast India Using Drought and Blast Linked Markers. Rice Science, 24(1), pp. 10–20. doi: 10.1016/j.rsci.2016.07.003.
Arisha, MH, Liang BK, Shah SNM, Gong ZH, Li DW. 2015. Kill Curve Analysis and Response of First Generation Capsicum annuum L. B12 Cultivar to Ethyl Methane Sulfonate. Frontiers in Plant Science.6(June). 1–11. doi: 10.3389/fpls.2015.00399.
Aslam K, Arif M. 2014. SSR Analysis of Chromosomes 3 and 7 of Rice (Oryza Staiva L.) Associated with Grain Length. Pakistan Journal of Botany. 46(4):1363–1373.
Aswaneeza, KA, Rawaidah N, Rodrigues KF, Cheong BE. 2015. Evaluation of the Mutagenic Effect of Ethyl Methanesulfonate on Hexokinase nuclear DNA Locus of Solanum lycopersicum. pp.88–93. In Proceeding of Int'l Conf. on Advances in Science, Engg. Technology & Natural Resources (ICASETNR-15) Aug. 27-28, 2015 Kota Kinabalu (Malaysia). doi: 10.15242/iicbe.c0815048.
Bahurupe JV, Sakhare SB, Kulwal PL, Akhare AA, Pawar, BD. 2018. Genetic Diversity Analysis in Chilli (Capsicum annuum L.) Found in Manipur Using RAPD Markers. International Journal of Current Microbiology and Applied Sciences. 7(10):257–262. doi: 10.20546/ijcmas.2018.710.027.
Bijalwan P. Mishra AC. 2017. Correlation and Path Coefficient Analysis in Bird’s Eye Chilli (Capsicum frutescens L.,) for Yield and Yield Attributing Traits. International Journal of Agricultural Science and Research. 7(3): 259–266. doi: 10.24247/ijasrjun201733.
Can, N.D., & Yoshida, T (1999). Genotypic and Phenotypic Variances and Covariance's in Early Maturing Grain Sorghum in a Double Cropping. Plant Production Science. 2(1):67-70. http://dx.doi.org/10.1626/pps.2.67.
Chattopadhyay A, Sharani AB, Nuka D, Subrata D. 2011. Diversity of Genetic Resources and Genetic Association Analyses of Green and Dry Chillies of Eastern India. Chilean Journal of Agricultural Research. 71(3):350–356. doi: 10.4067/S0718-58392011000300002.
Cheng J, Zhao Z, Li B, Qin C, Wu Z, Trejo-Saavedra DL, Luo X, Cui J, Rivera-Bustamante RF, Li S, Hu K. 2016. A Comprehensive Characterization of Simple Sequence Repeats in Pepper Genomes Provides Valuable Resources for Marker Development in Capsicum. Scientific Reports. 6:1–12. doi: 10.1038/srep18919.
Dhaliwal MS, Yadav A, Jindal SK. 2014. Molecular characterization and diversity analysis in chilli pepper using simple sequence repeats (SSR) markers. African Journal of Biotechnology. 13(31):3137–3143. doi: 10.5897/ajb2014.13695.
Falconer DS. 1989. Introduction to Quantitative Genetics. New York,: Ronald Press Co. doi: 10.2307/2985722.
Gandhi ES, Devi AS, Mullainathan L. 2014. The Effect of Ethyl Methane Sulphonate and Diethyl Sulphate on Chilli (Capsicum annuum L.) in M1 Generation. International Letters of Natural Sciences. 10:18–23. doi: 10.18052/www.scipress.com/ilns.10.18.
Geleta, LF, Labuschagne MT, Viljoen CD (2005) Genetic Variability in Pepper (Capsicum annuum L.) Estimated by Morphological Data and Amplified Fragment Length Polymorphism Markers. Biodiversity and Conservation. 14(10): 2361–2375. doi: 10.1007/s10531-004-1669-9.
Giarrocco LE, Marassi MA, Salerno GL. 2007. Assessment of the Genetic Diversity in Argentine Rice Cultivars with SSR Markers. Crop Science. 47(2): 853–860. doi: 10.2135/cropsci2005.07.0198.
Greene, EA. 2003. Spectrum of Chemically Induced Mutations from a Large-scale Reverse-genetic Screen in Arabidopsis. Genetics. 164(2):731–740.
Herrera, TG, Duque DP, Almeida IP, Núñez GT, Pieters AJ, Martinez CP, Tohme JM. 2008. Assessment of Genetic Diversity in Venezuelan Rice Cultivars Using Simple Sequence Sepeats Markers. Electronic Journal of Biotechnology. 11(5):1-14. doi: 10.2225/vol11-issue5-fulltext-6.
Hidayat, SH, Endang O, Manzila I, Sujiprihati S. 2012. Occurrence of Chili Veinal Mottle Virus (ChiVMV) in Indonesia and Response of Chili Germplasms to ChiVMV Infection. Journal of the International Society for Southeast Asian Agricultural Sciences. 18(2):55–61.
Kenyon, Kenyon L, Kumar S, Tsai WS, Hughes JD. 2014. Virus Diseases of Peppers (Capsicum spp.) and Their Control. Advances in Virus Research. 90(4):297–354. doi: 10.1016/B978-0-12-801246-8.00006-8.
Khalil F, Xiao N, Tayyab M, Cheng P. 2018. Screening of Ems-induced Drought-tolerant Sugarcane Mutants Employing Physiological, Molecular and Enzymatic Approaches. Agronomy. 8(226):1-13. doi: 10.3390/agronomy8100226.
Kwon YS, Kim KM, Kim DH, Eun MY, Sohn JK. 2002. Marker-assisted Introgression of Quantitative Trait Loci Associated with Plant Regeneration Ability in Anther Culture of Rice (Oryza sativa L.). Molecules and Cells. 14(1):24–28.
Liu K, Muse SV. 2005. PowerMaker: An Integrated Analysis Environment for Genetic Maker Analysis. Bioinformatics. 21(9): 2128–2129. doi: 10.1093/bioinformatics/bti282.
Manzila I, Gunaeni N, Kusandriani Y, Priyatno TP. 2015. Resistance and Phenotypic Character of Chili M 2 Mutant Lines Against Chilli Veinal Mottle Virus. Journal Agrobiogen. 11(2). 73–80.
Mariyono J, Bhattarai M. 2016. The Economic Aspects of Chilli Production in Central Java. Economic Journal of Emerging Markets. 8(2):85–97. doi: 10.20885/ejem.vol8.iss2.art1.
Jodi MY, Madalageri MB, Ganiger VM, Bhuvaneswari G, Patil HB, Kotikal YK. 2017. Character Association and Path Analysis Studies for Yield and Yield Attributes in Green Chilli (Capsicum annuum L.). Vegetos, 30(Special Issue 2):369–373. doi: 10.5958/2229-4473.2017.00100.8.
Nachimuthu VV, Muthurajan R, Duraialaguraja S, Sivakami R, Pandian BA, Ponniah G, Gunasekaran K, Swaminathan MKSK, Sabariappan R. 2015. Analysis of Population Structure and Genetic Diversity in Rice Germplasm Using SSR Markers: An Initiative Towards Association Mapping of Agronomic Traits in Oryza Sativa. Rice. Rice. 8(1):62-67. doi: 10.1186/s12284-015-0062-5.
Narayana SS, Singh P. 2015. Statistaical and biometrical techniques in plant breeding. 5th edition. New Delhi: Kalyani Publishers.
Nei M. 1973. Analysis of gene diversity in subdivided populations. Proceedings of the National Academy of Sciences of the United States of America. 70(12):3321–3323.
Olasupo FO, Ilori1 CO, Stanley EA, Owoeye TE, Igwe DO. 2018. Genetic Analysis of Selected Mutants of Cowpea (Vigna unguiculata [L.] Walp) Using Simple Sequence Repeat Markers. American Journal of Plant Sciences. 9(13): 2728–2756. doi: 10.4236/ajps.2018.913199.
Qosim, WA, Rachmadi M, Hamdani JS and Nuri I. 2013. Phenotypic Performance, Variability, and Heritability of 32 High Yielding Red Pepper Genotypes, Journal Agronomi Indonesia. 41(2):140-146.
Ramezani P, Siavozh M, More AD, Ebrahimi M, Dastan S. 2017. Gamma Rays and EMS Induced Flower Color Mutation in Grasspea (Lathyrus sativus Linn.). Tarim Bilimleri Dergisi. 23(4):423–427. doi: 10.15832/tbd.13090.
Rohlf FJ. 2017. NTSYSpc Numerical Taxonomy and Multivariance Analysis System, Version 2.02. Setauket, New York.
Sadat S, Hoveize M. 2012. Mutation Induction Using Ethyl Methanesulfonate (EMS) in Regenerated Plantlets of Two Varieties of Sugarcane CP48-103 and CP57-614. Africa Journal Agricultural Research. 7(8):1282–1288. doi: 10.5897/AJAR11.1345.
Salem KFM, Sallam A. 2016. Analysis of Population Structure and Genetic diversity of Egyptian and Exotic Rice (Oryza sativa L.) Genotypes. Comptes Rendus - Biologies. 339(1):1–9. doi: 10.1016/j.crvi.2015.11.003.
Schulman AH. 2007. Molecular Markers to Assess Genetic Diversity. Euphytica. 158(3):313–321. doi: 10.1007/s10681-006-9282-5.
Shalaby TA, El-Banna A. 2013. Molecular and Horticultural Characteristics of In vitro Induced Tomato Mutants. Journal of Agricultural Science. 5(10):155–163. doi: 10.5539/jas.v5n10p155.
Sharmin A, Ekramu H, Masudul H, Fahima K. 2018. Molecular Diversity Analysis of Some Chilli (Capsicum spp.) Genotypes Using SSR Markers. American Journal of Plant Sciences. 9(03): 368–379. doi: 10.4236/ajps.2018.93029.
Shah H, Khalid S, Ahmad I, 2001. Prevalence and Distribution of Four Pepper Viruses in Sindh, Punjab and North-West Frontier Province. Online Journal of Biological Sciences. 1: 214-217.
Smith JSC, Chin ECL, Shu H, Smith OS, Wall SJ, Senior ML, Mitchell SE, Kresovich S, Ziegle J. 1997. An Evaluation of the Utility of SSR Loci as Molecular Markers in Maize (Zea mays L.): Comparisons with Data from RFLPS and Pedigree. Theoretical and Applied Genetics. 95(1–2):163–173. doi: 10.1007/s001220050544.
Sreelathakumary I, Rajamony L. 2004. Variability, Heritability and Genetic Advance in Chilli (Capsicum annuum L.). Journal of Tropical Agriculture. 42(1–2):35–37.
Till BJ, Cooper J, Ti TH, Colowit P, Greene EA. 2007. Discovery of Chemically Induced Mutations in Rice by TILLING. BMC Plant Biology. 7: 1–12. doi: 10.1186/1471-2229-7-19.
Tsai, H, Howell T, Nitcher R, Missirian V, Watson B, Ngo KJ, Lieberman M, Fass J, Uauy C, Tran RK, Khan AA, Filkov V, Tai TH, Dubcovsky J, Comai L (2011) ‘Discovery of rare mutations in populations: Tilling by sequencing’, Plant Physiology, 156(3), pp. 1257–1268. doi: 10.1104/pp.110.169748.
Uchida, N, Sakamoto T, Kurata T, Tasaka M. 2011. Identification of EMS-induced Causal Mutations in a Non-reference Arabidopsis thaliana Accession by Whole Genome Sequencing. Plant and Cell Physiology, 52(4): 716–722. doi: 10.1093/pcp/pcr029.
Yadeta B, Belew D, Gebreselassie W, Marame F. 2011. Variability, Heritability and Genetic Advance in Hot Pepper (Capsicum annuum L.) Genotypes in West Shoa Ethiopia. American-Eurasian Journal of Agricultural & Environmental Sciences. 10: 587–592
Zalapa J, Cuevas H, Zhu H, Steffan S, Senalik D. 2012. Using Next-generation Sequencing Approaches to Isolate Simple Sequence Repeat (SSR) Loci in the Plant Sciences. American Journal Botany. 99:193–208.
Ajjapplavara PS, Patil SS, Hosmani RM, Patil AA, Gangaprasad S. 2005. Correlation and Path Coefficient Analysis in Chilli. Karnataka J.Agric.Sci.
Alonso, J. M. Stepanova AN, Leisse TJ, Kim CJ, Chen HP. 2003. Genome-wide Insertional Mutagenesis of Arabidopsis thaliana. Science. 301(5633):653–657. doi: 10.1126/science.1086391.
Alvarez A, Fuentes JL, Puldón V, Gómez PJ, Mora L, Duque MC, Gallego G, Tohme JM. 2007. Genetic Diversity Analysis of Cuban Traditional Rice (Oryza sativa L.) Varieties Based on Microsatellite Markers. Genetics and Molecular Biology. 30(4): 1109–1117. doi: 10.1590/S1415-47572007000600014.
Anh TTT, Tran DK, Tran DD, Tran DX. 2018. Identification of Phenotypic Variation and Genetic Diversity in Rice (Oryza sativa L.) Mutants. Agriculture (Switzerland). doi: 10.3390/agriculture8020030.
Anupam A, Imam J, Quatadah SM, Siddaiah A, Das SP, Variar M, Mandal NP. 2017. Genetic Diversity Analysis of Rice Germplasm in Tripura State of Northeast India Using Drought and Blast Linked Markers. Rice Science, 24(1), pp. 10–20. doi: 10.1016/j.rsci.2016.07.003.
Arisha, MH, Liang BK, Shah SNM, Gong ZH, Li DW. 2015. Kill Curve Analysis and Response of First Generation Capsicum annuum L. B12 Cultivar to Ethyl Methane Sulfonate. Frontiers in Plant Science.6(June). 1–11. doi: 10.3389/fpls.2015.00399.
Aslam K, Arif M. 2014. SSR Analysis of Chromosomes 3 and 7 of Rice (Oryza Staiva L.) Associated with Grain Length. Pakistan Journal of Botany. 46(4):1363–1373.
Aswaneeza, KA, Rawaidah N, Rodrigues KF, Cheong BE. 2015. Evaluation of the Mutagenic Effect of Ethyl Methanesulfonate on Hexokinase nuclear DNA Locus of Solanum lycopersicum. pp.88–93. In Proceeding of Int'l Conf. on Advances in Science, Engg. Technology & Natural Resources (ICASETNR-15) Aug. 27-28, 2015 Kota Kinabalu (Malaysia). doi: 10.15242/iicbe.c0815048.
Bahurupe JV, Sakhare SB, Kulwal PL, Akhare AA, Pawar, BD. 2018. Genetic Diversity Analysis in Chilli (Capsicum annuum L.) Found in Manipur Using RAPD Markers. International Journal of Current Microbiology and Applied Sciences. 7(10):257–262. doi: 10.20546/ijcmas.2018.710.027.
Bijalwan P. Mishra AC. 2017. Correlation and Path Coefficient Analysis in Bird’s Eye Chilli (Capsicum frutescens L.,) for Yield and Yield Attributing Traits. International Journal of Agricultural Science and Research. 7(3): 259–266. doi: 10.24247/ijasrjun201733.
Can, N.D., & Yoshida, T (1999). Genotypic and Phenotypic Variances and Covariance's in Early Maturing Grain Sorghum in a Double Cropping. Plant Production Science. 2(1):67-70. http://dx.doi.org/10.1626/pps.2.67.
Chattopadhyay A, Sharani AB, Nuka D, Subrata D. 2011. Diversity of Genetic Resources and Genetic Association Analyses of Green and Dry Chillies of Eastern India. Chilean Journal of Agricultural Research. 71(3):350–356. doi: 10.4067/S0718-58392011000300002.
Cheng J, Zhao Z, Li B, Qin C, Wu Z, Trejo-Saavedra DL, Luo X, Cui J, Rivera-Bustamante RF, Li S, Hu K. 2016. A Comprehensive Characterization of Simple Sequence Repeats in Pepper Genomes Provides Valuable Resources for Marker Development in Capsicum. Scientific Reports. 6:1–12. doi: 10.1038/srep18919.
Dhaliwal MS, Yadav A, Jindal SK. 2014. Molecular characterization and diversity analysis in chilli pepper using simple sequence repeats (SSR) markers. African Journal of Biotechnology. 13(31):3137–3143. doi: 10.5897/ajb2014.13695.
Falconer DS. 1989. Introduction to Quantitative Genetics. New York,: Ronald Press Co. doi: 10.2307/2985722.
Gandhi ES, Devi AS, Mullainathan L. 2014. The Effect of Ethyl Methane Sulphonate and Diethyl Sulphate on Chilli (Capsicum annuum L.) in M1 Generation. International Letters of Natural Sciences. 10:18–23. doi: 10.18052/www.scipress.com/ilns.10.18.
Geleta, LF, Labuschagne MT, Viljoen CD (2005) Genetic Variability in Pepper (Capsicum annuum L.) Estimated by Morphological Data and Amplified Fragment Length Polymorphism Markers. Biodiversity and Conservation. 14(10): 2361–2375. doi: 10.1007/s10531-004-1669-9.
Giarrocco LE, Marassi MA, Salerno GL. 2007. Assessment of the Genetic Diversity in Argentine Rice Cultivars with SSR Markers. Crop Science. 47(2): 853–860. doi: 10.2135/cropsci2005.07.0198.
Greene, EA. 2003. Spectrum of Chemically Induced Mutations from a Large-scale Reverse-genetic Screen in Arabidopsis. Genetics. 164(2):731–740.
Herrera, TG, Duque DP, Almeida IP, Núñez GT, Pieters AJ, Martinez CP, Tohme JM. 2008. Assessment of Genetic Diversity in Venezuelan Rice Cultivars Using Simple Sequence Sepeats Markers. Electronic Journal of Biotechnology. 11(5):1-14. doi: 10.2225/vol11-issue5-fulltext-6.
Hidayat, SH, Endang O, Manzila I, Sujiprihati S. 2012. Occurrence of Chili Veinal Mottle Virus (ChiVMV) in Indonesia and Response of Chili Germplasms to ChiVMV Infection. Journal of the International Society for Southeast Asian Agricultural Sciences. 18(2):55–61.
Kenyon, Kenyon L, Kumar S, Tsai WS, Hughes JD. 2014. Virus Diseases of Peppers (Capsicum spp.) and Their Control. Advances in Virus Research. 90(4):297–354. doi: 10.1016/B978-0-12-801246-8.00006-8.
Khalil F, Xiao N, Tayyab M, Cheng P. 2018. Screening of Ems-induced Drought-tolerant Sugarcane Mutants Employing Physiological, Molecular and Enzymatic Approaches. Agronomy. 8(226):1-13. doi: 10.3390/agronomy8100226.
Kwon YS, Kim KM, Kim DH, Eun MY, Sohn JK. 2002. Marker-assisted Introgression of Quantitative Trait Loci Associated with Plant Regeneration Ability in Anther Culture of Rice (Oryza sativa L.). Molecules and Cells. 14(1):24–28.
Liu K, Muse SV. 2005. PowerMaker: An Integrated Analysis Environment for Genetic Maker Analysis. Bioinformatics. 21(9): 2128–2129. doi: 10.1093/bioinformatics/bti282.
Manzila I, Gunaeni N, Kusandriani Y, Priyatno TP. 2015. Resistance and Phenotypic Character of Chili M 2 Mutant Lines Against Chilli Veinal Mottle Virus. Journal Agrobiogen. 11(2). 73–80.
Mariyono J, Bhattarai M. 2016. The Economic Aspects of Chilli Production in Central Java. Economic Journal of Emerging Markets. 8(2):85–97. doi: 10.20885/ejem.vol8.iss2.art1.
Jodi MY, Madalageri MB, Ganiger VM, Bhuvaneswari G, Patil HB, Kotikal YK. 2017. Character Association and Path Analysis Studies for Yield and Yield Attributes in Green Chilli (Capsicum annuum L.). Vegetos, 30(Special Issue 2):369–373. doi: 10.5958/2229-4473.2017.00100.8.
Nachimuthu VV, Muthurajan R, Duraialaguraja S, Sivakami R, Pandian BA, Ponniah G, Gunasekaran K, Swaminathan MKSK, Sabariappan R. 2015. Analysis of Population Structure and Genetic Diversity in Rice Germplasm Using SSR Markers: An Initiative Towards Association Mapping of Agronomic Traits in Oryza Sativa. Rice. Rice. 8(1):62-67. doi: 10.1186/s12284-015-0062-5.
Narayana SS, Singh P. 2015. Statistaical and biometrical techniques in plant breeding. 5th edition. New Delhi: Kalyani Publishers.
Nei M. 1973. Analysis of gene diversity in subdivided populations. Proceedings of the National Academy of Sciences of the United States of America. 70(12):3321–3323.
Olasupo FO, Ilori1 CO, Stanley EA, Owoeye TE, Igwe DO. 2018. Genetic Analysis of Selected Mutants of Cowpea (Vigna unguiculata [L.] Walp) Using Simple Sequence Repeat Markers. American Journal of Plant Sciences. 9(13): 2728–2756. doi: 10.4236/ajps.2018.913199.
Qosim, WA, Rachmadi M, Hamdani JS and Nuri I. 2013. Phenotypic Performance, Variability, and Heritability of 32 High Yielding Red Pepper Genotypes, Journal Agronomi Indonesia. 41(2):140-146.
Ramezani P, Siavozh M, More AD, Ebrahimi M, Dastan S. 2017. Gamma Rays and EMS Induced Flower Color Mutation in Grasspea (Lathyrus sativus Linn.). Tarim Bilimleri Dergisi. 23(4):423–427. doi: 10.15832/tbd.13090.
Rohlf FJ. 2017. NTSYSpc Numerical Taxonomy and Multivariance Analysis System, Version 2.02. Setauket, New York.
Sadat S, Hoveize M. 2012. Mutation Induction Using Ethyl Methanesulfonate (EMS) in Regenerated Plantlets of Two Varieties of Sugarcane CP48-103 and CP57-614. Africa Journal Agricultural Research. 7(8):1282–1288. doi: 10.5897/AJAR11.1345.
Salem KFM, Sallam A. 2016. Analysis of Population Structure and Genetic diversity of Egyptian and Exotic Rice (Oryza sativa L.) Genotypes. Comptes Rendus - Biologies. 339(1):1–9. doi: 10.1016/j.crvi.2015.11.003.
Schulman AH. 2007. Molecular Markers to Assess Genetic Diversity. Euphytica. 158(3):313–321. doi: 10.1007/s10681-006-9282-5.
Shalaby TA, El-Banna A. 2013. Molecular and Horticultural Characteristics of In vitro Induced Tomato Mutants. Journal of Agricultural Science. 5(10):155–163. doi: 10.5539/jas.v5n10p155.
Sharmin A, Ekramu H, Masudul H, Fahima K. 2018. Molecular Diversity Analysis of Some Chilli (Capsicum spp.) Genotypes Using SSR Markers. American Journal of Plant Sciences. 9(03): 368–379. doi: 10.4236/ajps.2018.93029.
Shah H, Khalid S, Ahmad I, 2001. Prevalence and Distribution of Four Pepper Viruses in Sindh, Punjab and North-West Frontier Province. Online Journal of Biological Sciences. 1: 214-217.
Smith JSC, Chin ECL, Shu H, Smith OS, Wall SJ, Senior ML, Mitchell SE, Kresovich S, Ziegle J. 1997. An Evaluation of the Utility of SSR Loci as Molecular Markers in Maize (Zea mays L.): Comparisons with Data from RFLPS and Pedigree. Theoretical and Applied Genetics. 95(1–2):163–173. doi: 10.1007/s001220050544.
Sreelathakumary I, Rajamony L. 2004. Variability, Heritability and Genetic Advance in Chilli (Capsicum annuum L.). Journal of Tropical Agriculture. 42(1–2):35–37.
Till BJ, Cooper J, Ti TH, Colowit P, Greene EA. 2007. Discovery of Chemically Induced Mutations in Rice by TILLING. BMC Plant Biology. 7: 1–12. doi: 10.1186/1471-2229-7-19.
Tsai, H, Howell T, Nitcher R, Missirian V, Watson B, Ngo KJ, Lieberman M, Fass J, Uauy C, Tran RK, Khan AA, Filkov V, Tai TH, Dubcovsky J, Comai L (2011) ‘Discovery of rare mutations in populations: Tilling by sequencing’, Plant Physiology, 156(3), pp. 1257–1268. doi: 10.1104/pp.110.169748.
Uchida, N, Sakamoto T, Kurata T, Tasaka M. 2011. Identification of EMS-induced Causal Mutations in a Non-reference Arabidopsis thaliana Accession by Whole Genome Sequencing. Plant and Cell Physiology, 52(4): 716–722. doi: 10.1093/pcp/pcr029.
Yadeta B, Belew D, Gebreselassie W, Marame F. 2011. Variability, Heritability and Genetic Advance in Hot Pepper (Capsicum annuum L.) Genotypes in West Shoa Ethiopia. American-Eurasian Journal of Agricultural & Environmental Sciences. 10: 587–592
Zalapa J, Cuevas H, Zhu H, Steffan S, Senalik D. 2012. Using Next-generation Sequencing Approaches to Isolate Simple Sequence Repeat (SSR) Loci in the Plant Sciences. American Journal Botany. 99:193–208.
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