Free Standard AU & NZ Shipping For All Book Orders Over $80!
Register      Login
Crop and Pasture Science Crop and Pasture Science Society
Plant sciences, sustainable farming systems and food quality
RESEARCH ARTICLE

Development and validation of microsatellite markers for kikuyu grass using next generation sequencing technology

Juliana Arango https://orcid.org/0000-0002-4623-4588 A * , Albeiro López A , Edna Márquez B and Julián Echeverri A
+ Author Affiliations
- Author Affiliations

A Grupo de Investigación en Biodiversidad y Genética Molecular-BIOGEM [Research Group on Biodiversity and Molecular Genetics-BIOGEM], Universidad Nacional de Colombia, sede Medellín 050001, Colombia.

B Facultad de Ciencias, Escuela de Biociencias, Universidad Nacional de Colombia, sede Medellín 050001, Colombia.

* Correspondence to: jarangog@unal.edu.co

Handling Editor: Rajeev Varshney

Crop & Pasture Science 73(4) 415-424 https://doi.org/10.1071/CP21331
Submitted: 13 May 2021  Accepted: 17 November 2021   Published: 7 February 2022

© 2022 The Author(s) (or their employer(s)). Published by CSIRO Publishing

Abstract

Context: The kikuyu grass (Cenchrus clandestinus) is native to Africa and is one of the most widely used grasses for forage feeding in dairy regions around the world.

Aims: To obtain the first set of microsatellite markers for the population genetics study of kikuyu grass, using nextgeneration sequencing technology (Illumina MiSeq).

Methods: Sixty loci were evaluated, in which a subset of 12 loci were selected to be used for a complete population analysis in 108 samples of kikuyu, and were grouped in to three zones of Colombia. The three approaches with which the genetic structure was evaluated.

Key results: Obtained same tendency of grouping reflects a low genetic differentiation, specifically evidencing differences between the northern zone of Antioquia and the zones that comprise the other territories of Colombia.

Conclusions: These reads of microsatellite loci help to complement the information on the genetic structure of the populations of the kikuyu, and will be useful for the characterisation and evaluation of the diversity of germplasm in other parts of the world.

Implications: The set of microsatellite markers developed has a species-specific reproducibility and could be used for studies in other Cenchrus individuals and particularly in future investigations with Kikuyu grass. Likewise, this research presents findings in a broad context and relates them to other pasture species.

Keywords: Cenchrus clandestinus, clonal genotype detection, genetic diversity, genetic structure, kikuyu grass, microsatellite markers, molecular marker, next generation sequencing, population genetics.


References

Balan D, Jaramillo M, Restrepo LM, Saglimbeni S (2014) Ceba de ganado Angus en trópico alto con kikuyu (Pennisetum clandestinum Ex Chiov.). Available at http://bdigital.ces.edu.co:8080/repositorio/bitstream/10946/3874/1/Ceba_Ganado_Angus.pdf

Blacket MJ, Robin C, Good RT, Lee SF, Miller AD (2012) Universal primers for fluorescent labelling of PCR fragments—an efficient and cost-effective approach to genotyping by fluorescence. Molecular Ecology Resources 12, 456–463.
Universal primers for fluorescent labelling of PCR fragments—an efficient and cost-effective approach to genotyping by fluorescence.Crossref | GoogleScholarGoogle Scholar | 22268566PubMed |

Bolger AM, Lohse M, Usadel B (2014) Trimmomatic: a flexible trimmer for Illumina sequence data. Bioinformatics 30, 2114–2120.
Trimmomatic: a flexible trimmer for Illumina sequence data.Crossref | GoogleScholarGoogle Scholar | 24695404PubMed |

Bourke CA (2007) A review of kikuyu grass (Pennisetum clandestinum) poisoning in cattle. Australian Veterinary Journal 85, 261–267.
A review of kikuyu grass (Pennisetum clandestinum) poisoning in cattle.Crossref | GoogleScholarGoogle Scholar | 17615037PubMed |

Castoe TA, Poole AW, Gu W, Jason de Koning AP, Daza JM, Smith EN, Pollock DD (2010) Rapid identification of thousands of copperhead snake (Agkistrodon contortrix) microsatellite loci from modest amounts of 454 shotgun genome sequence. Molecular Ecology Resources 10, 341–347.
Rapid identification of thousands of copperhead snake (Agkistrodon contortrix) microsatellite loci from modest amounts of 454 shotgun genome sequence.Crossref | GoogleScholarGoogle Scholar | 21565030PubMed |

Clark AG (1993) Evolutionary inferences from molecular characterization of self-incompatibility alleles. In ‘Mechanisms of molecular evolution: introduction to molecular paleopopulation biology’. (Eds AC Takahata, AG Clark) pp. 79–108. (Sinauer Associates: Sunderland, MA)

Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conservation Genetics Resources 4, 359–361.
STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method.Crossref | GoogleScholarGoogle Scholar |

Ellstrand NC, Roose ML (1987) Patterns of genotypic diversity in clonal plant species. American Journal of Botany 74, 123–131.
Patterns of genotypic diversity in clonal plant species.Crossref | GoogleScholarGoogle Scholar |

Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology 14, 2611–2620.
Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study.Crossref | GoogleScholarGoogle Scholar | 15969739PubMed |

Excoffier L, Laval G, Schneider S (2005) Arlequin (version 3.0): an integrated software package for population genetics data analysis. Evolutionary Bioinformatics 1, 47–50.
Arlequin (version 3.0): an integrated software package for population genetics data analysis.Crossref | GoogleScholarGoogle Scholar |

Fukumoto GK, Lee CN (2003) Kikuyu grass for forage. Livestock Management 5. (Cooperative Extension Service, College of tropical agriculture and human resources, University of Hawaii at Manoa). Available at https://www.ctahr.hawaii.edu/oc/freepubs/pdf/LM-5.pdf

García SC, Islam MR, Clark CEF, Martin PM (2014) Kikuyu-based pasture for dairy production: a review. Crop and Pasture Science 65, 787–797.
Kikuyu-based pasture for dairy production: a review.Crossref | GoogleScholarGoogle Scholar |

Hartl DL, Clark AG (2006) ‘Principles of population genetics.’ (Sinauer Associates: Sunderland)

Holton TA, Skabo SJ, Lowe KF, Sinclair K (2007) Genetic fingerprinting of natural kikuyu populations in Australia. Tropical Grasslands 41, 236–237.

Insuasty E, Apráez J, Navia J (2011) Efecto del arreglo silvopastoril aliso (Alnus Acuminata K.) y kikuyu (Pennisetum Clandestinum H.) sobre el comportamiento productivo en novillas Holstein en el altiplano del departamento de Nariño. Revista Agroforestería Neotropical 1, 1–8.

Jakobsson M, Rosenberg NA (2007) CLUMPP: a cluster matching and permutation program for dealing with label switching and multimodality in analysis of population structure. Bioinformatics 23, 1801–1806.
CLUMPP: a cluster matching and permutation program for dealing with label switching and multimodality in analysis of population structure.Crossref | GoogleScholarGoogle Scholar | 17485429PubMed |

Jombart T (2008) adegenet: a R package for the multivariate analysis of genetic markers. Bioinformatics 24, 1403–1405.
adegenet: a R package for the multivariate analysis of genetic markers.Crossref | GoogleScholarGoogle Scholar | 18397895PubMed |

Jungmann L, Vigna BBZ, Boldrini KR, Sousa ACB, Do Valle CB, Resende RMS, Pagliarini MS, Zucchi MI, De Souza AP (2010) Genetic diversity and population structure analysis of the tropical pasture grass Brachiaria humidicola based on microsatellites, cytogenetics, morphological traits, and geographical origin. Genome 53, 698–709.
Genetic diversity and population structure analysis of the tropical pasture grass Brachiaria humidicola based on microsatellites, cytogenetics, morphological traits, and geographical origin.Crossref | GoogleScholarGoogle Scholar | 20924419PubMed |

Kandel R, Singh HP, Singh BP, Harris-Shultz KR, Anderson WF (2016) Assessment of Genetic Diversity in Napier Grass (Pennisetum purpureum Schum.) using Microsatellite, Single-Nucleotide Polymorphism and Insertion-Deletion Markers from Pearl Millet (Pennisetum glaucum [L.] R. Br.). Plant Molecular Biology Reporter 34, 265–272.
Assessment of Genetic Diversity in Napier Grass (Pennisetum purpureum Schum.) using Microsatellite, Single-Nucleotide Polymorphism and Insertion-Deletion Markers from Pearl Millet (Pennisetum glaucum [L.] R. Br.).Crossref | GoogleScholarGoogle Scholar |

Khumalo TP (2015) Genetic identification of kikuyu grass (Pennisetum clandestinum) cultivars by RAPD and ISSR techniques. MSc thesis, The University of Kwazulu-Natal, South Africa.

Landínez-García RM, Márquez EJ (2016) Development and characterization of 24 polymorphic microsatellite loci for the freshwater fish Ichthyoelephas longirostris (Characiformes: Prochilodontidae). PeerJ 4, e2419
Development and characterization of 24 polymorphic microsatellite loci for the freshwater fish Ichthyoelephas longirostris (Characiformes: Prochilodontidae).Crossref | GoogleScholarGoogle Scholar | 27635363PubMed |

Lowe KF, Bowdler TM, Sinclair K, Holton TA, Skabo SJ (2010) Phenotypic and genotypic variation within populations of kikuyu (Pennisetum clandestinum) in Australia. Tropical Grasslands 44, 84–94.

Marais J (2001) Factors affecting the nutritive value of kikuyu grass (Pennisetum clandestinum)—a review. Tropical Grasslands 35, 65–84.

Mariac C, Luong V, Kapran I, Mamadou A, Sagnard F, Deu M, Chantereau J, Gerard B, Ndjeunga J, Bezançon G, Pham J-L, Vigouroux Y (2006) Diversity of wild and cultivated pearl millet accessions (Pennisetum glaucum [L.] R. Br.) in Niger assessed by microsatellite markers. Theoretical and Applied Genetics 114, 49–58.
Diversity of wild and cultivated pearl millet accessions (Pennisetum glaucum [L.] R. Br.) in Niger assessed by microsatellite markers.Crossref | GoogleScholarGoogle Scholar | 17047913PubMed |

Márquez Girón SM, Mosquera Ballesteros R, Herrera Torres M, Monedero C (2010) Estudio de la absorción y distribución del clorpirifos en plantas de pasto kikuyu cultivadas hidropónicamente. Revista Colombiana de Ciencias Pecuarias 23, 158–165.

Matsuoka Y, Vigouroux Y, Goodman MM, Sanchez JG, Buckler E, Doebley J (2002) A single domestication for maize shown by multilocus microsatellite genotyping. Proceedings of the National Academy of Sciences of the United States of America 99, 6080–6084.
A single domestication for maize shown by multilocus microsatellite genotyping.Crossref | GoogleScholarGoogle Scholar | 11983901PubMed |

McKey D, Elias M, Pujol B, Duputié A (2010) The evolutionary ecology of clonally propagated domesticated plants. New Phytologist 186, 318–332.
The evolutionary ecology of clonally propagated domesticated plants.Crossref | GoogleScholarGoogle Scholar |

Mears PT (1970) Kikuyu—(Pennisetum clandestinum) as a pasture grass—a review. Tropical Grasslands 4, 139–152.

Meirmans PG (2006) Using the AMOVA framework to estimate a standardized genetic differentiation measure. Evolution 60, 2399
Using the AMOVA framework to estimate a standardized genetic differentiation measure.Crossref | GoogleScholarGoogle Scholar | 17236430PubMed |

Meirmans PG, Hedrick PW (2011) Assessing population structure: FST and related measures. Molecular Ecology Resources 11, 5–18.
Assessing population structure: FST and related measures.Crossref | GoogleScholarGoogle Scholar | 21429096PubMed |

Miyasaka SC, Hansen JD, Fukumoto GK (2007) Resistance to yellow sugarcane aphid: screening kikuyu and other grasses. Crop Protection 26, 503–510.
Resistance to yellow sugarcane aphid: screening kikuyu and other grasses.Crossref | GoogleScholarGoogle Scholar |

Mock J (2016) Management and genetic variability of kikuyugrass (Pennisetum clandestinum Hochst. Ex Chiov.). University of California Riverside.

Morris B (2009) Variation and breeding of kikuyu grass (Pennisetum clandestinum). PhD thesis, Plant Breeding Institute, The University of Sydney, NSW, Australia.

Muscolo A, Panuccio MR, Eshel A (2013) Ecophysiology of Pennisetum clandestinum: a valuable salt tolerant grass. Environmental and Experimental Botany 92, 55–63.
Ecophysiology of Pennisetum clandestinum: a valuable salt tolerant grass.Crossref | GoogleScholarGoogle Scholar |

Ozias-Akins P, Akiyama Y, Hanna WW (2003) Molecular characterization of the genomic region linked with apomixis in Pennisetum/Cenchrus. Functional & Integrative Genomics 3, 94–104.
Molecular characterization of the genomic region linked with apomixis in Pennisetum/Cenchrus.Crossref | GoogleScholarGoogle Scholar |

Parker D (1941) Strain variation and seed production in kikuyu gras (Pennisetum clandestinum Hochst.). Journal of Agriculture of South Australia 45, 55–59.

Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update. Bioinformatics 28, 2537–2539.
GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research—an update.Crossref | GoogleScholarGoogle Scholar |

Pessoa-Filho M, Azevedo ALS, Sobrinho FS, Gouvea EG, Martins AM, Ferreira ME (2015) Genetic diversity and structure of ruzigrass germplasm collected in Africa and Brazil. Crop Science 55, 2736–2745.
Genetic diversity and structure of ruzigrass germplasm collected in Africa and Brazil.Crossref | GoogleScholarGoogle Scholar |

Poulin J, Weller SG, Sakai AK (2005) Genetic diversity does not affect the invasiveness of fountain grass (Pennisetum setaceum) in Arizona, California and Hawaii. Diversity and Distributions 11, 241–247.
Genetic diversity does not affect the invasiveness of fountain grass (Pennisetum setaceum) in Arizona, California and Hawaii.Crossref | GoogleScholarGoogle Scholar |

Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155, 945–959.
Inference of population structure using multilocus genotype data.Crossref | GoogleScholarGoogle Scholar | 10835412PubMed |

Rice WR (1989) Analyzing tables of statistical tests. Evolution 43, 223–225.
Analyzing tables of statistical tests.Crossref | GoogleScholarGoogle Scholar | 28568501PubMed |

Rosenberg NA (2004) DISTRUCT: a program for the graphical display of population structure. Molecular Ecology Notes 4, 137–138.
DISTRUCT: a program for the graphical display of population structure.Crossref | GoogleScholarGoogle Scholar |

Rotmistrovsky K, Jang W, Schuler GD (2004) A web server for performing electronic PCR. Nucleic Acids Research 32, W108–W112.
A web server for performing electronic PCR.Crossref | GoogleScholarGoogle Scholar | 15215361PubMed |

Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. In ‘Methods and protocols: methods in molecular biology. Vol. 132’. (Eds S Krawetz, S Misener) pp. 365–386.
| Crossref |

Sambrook J, Russell D (2001) ‘Molecular cloning: a laboratory manual.’, 3rd edn. (Cold Spring Harbor Laboratory Press: New York)

Schmieder R, Edwards R (2011) Quality control and preprocessing of metagenomic datasets. Bioinformatics 27, 863–864.
Quality control and preprocessing of metagenomic datasets.Crossref | GoogleScholarGoogle Scholar | 21278185PubMed |

Selkoe KA, Toonen RJ (2006) Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers. Ecology Letters 9, 615–629.
Microsatellites for ecologists: a practical guide to using and evaluating microsatellite markers.Crossref | GoogleScholarGoogle Scholar | 16643306PubMed |

Silva PIT, Martins AM, Gouvea EG, Pessoa-Filho M, Ferreira ME (2013) Development and validation of microsatellite markers for Brachiaria ruziziensis obtained by partial genome assembly of Illumina single-end reads. BMC Genomics 14, 17
Development and validation of microsatellite markers for Brachiaria ruziziensis obtained by partial genome assembly of Illumina single-end reads.Crossref | GoogleScholarGoogle Scholar |

Tamiru M, Yamanaka S, Mitsuoka C, Babil P, Takagi H, Lopez-Montes A, Sartie A, Asiedu R, Terauchi R (2015) Development of genomic simple sequence repeat markers for yam. Crop Science 55, 2191–2200.
Development of genomic simple sequence repeat markers for yam.Crossref | GoogleScholarGoogle Scholar |

Van Oosterhout C, Hutchinson WF, Wills DPM, Shipley P (2004) MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data. Molecular Ecology Notes 4, 535–538.
MICRO-CHECKER: software for identifying and correcting genotyping errors in microsatellite data.Crossref | GoogleScholarGoogle Scholar |

Wilen CA, Holt JS, Ellstrand NC, Shaw RG, Science W, Wilen CA, Holt JS, Ellstrand NC, Shaw RG (1995) Genotypic diversity of kikuyugrass (Pennisetum clandestinum) populations in California. Weed Scienc 43, 209–214.
Genotypic diversity of kikuyugrass (Pennisetum clandestinum) populations in California.Crossref | GoogleScholarGoogle Scholar |

Zalapa JE, Cuevas H, Zhu H, Steffan S, Senalik D, Zeldin E, McCown B, Harbut R, Simon P (2012) Using next-generation sequencing approaches to isolate simple sequence repeat (SSR) loci in the plant sciences. American Journal of Botany 99, 193–208.
Using next-generation sequencing approaches to isolate simple sequence repeat (SSR) loci in the plant sciences.Crossref | GoogleScholarGoogle Scholar | 22186186PubMed |