Genetic diversity and phylogeny of ITS regions in sweet orange genotypes from San Luis Potosí
DOI:
https://doi.org/10.29312/remexca.v17i2.3956Keywords:
Citrus sinensis, Genetic variability, SequencingAbstract
The sweet orange is grown in several states of Mexico. Several research centers have citrus germplasm banks, including sweet oranges for genetic improvement. This work aimed to evaluate genetic diversity and perform a phylogenetic analysis based on the internal transcribed spacer (ITS) region of six sweet orange genotypes grown in San Luis Potosí, Mexico. In 2023, leaves of six varieties of sweet orange, known in the region as Sangre de Toro and Valencia, were collected. Genomic DNA was extracted, and ITS regions were amplified by PCR and sequenced. The percentage of identity and genetic distances were determined and a maximum-likelihood phylogenetic tree was generated. The results indicated that the intraspecific variability among C. sinensis genotypes in the ITS region is low to moderate (0-0.07). Particularly, among the population of Citrus sinensis (Sangre de Toro and Valencia varieties) from the regions of Axtla de Terrazas, Chimimexco-Tampacán and El Frijolillo-San Martín in San Luis Potosí, no differences were detected in their ITS regions. It is important to note that the number of samples analyzed was limited; therefore, it is necessary to evaluate a larger set of samples and varieties to obtain more robust conclusions regarding genetic variability in Citrus sinensis in Mexico. The phylogenetic analysis grouped the genotypes of Mexico into group G1, confirming the origin of the varieties with Asian countries (China, Korea and Vietnam), with which the percentage of identity was 100%.
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Abbaszadeh, M.; Sheidai, M.; Koohdar, F. and Shafieizargar, A. 2023. Notes on Sweet orange (Citrus sinensis L. Osbeck) populations’ divergence: landscape genetics, comparative phylogeny and Niche modeling. Preprint, Research Square. https://doi.org/10.21203/rs.3.rs-3801400/v1.
Ahsan, M. U.; Liu, Q.; Fang, L. and Wang, K. 2021. NanoCaller for accurate detection of SNPs and indels in difficult-to-map regions from long-read sequencing by haplotype-aware deep neural networks. Genome Biology. 22(1):1-33. https://doi.org/10.1186/s13059-021-02472-2.
Almeyda-León, I. H.; Narvaéz-Rodríguez, Á. I.; Pecina-Quintero, V.; Álvarez-Ojeda, M. G. y Núñez-Colín, C. A. 2023. Caracterización y diversidad genética de cítricos del banco de germoplasma del Campo Experimental General Terán. Biotecnología y Sustentabilidad. 8(1):26-41. https://doi.org/10.57737/biotecnologiaysust.v8i1.2276.
Almeyda-León, I. H.; Pecina-Quintero, V.; Álvarez-Ojeda, M. G.; Rodríguez-Guerra, R.; Acosta-Díaz, E. y Núñez-Colín, C. A. 2022. Caracterización y diversidad genética de naranjos dulces (Citrus sinensis L.) del banco de germoplasma del Campo Experimental General Terán. Biotecnología y Sustentabilidad. 7(1):80-95. https://doi.org/10.57737/biotecnologiaysust.v7i1.1645.
Alvarez, I. y Wendel, J. F. 2003. Ribosomal ITS sequences and plant phylogenetic inference. Molecular Phylogenetics and Evolution. 29(3):417-434. https://doi.org/10.1016/s1055-7903(03)00208-2.
Bermúdez-Guzmán, M. J.; Guzmán-González, S.; Orozco-Santos, M.; Velázquez-Monreal, J. J.; Buenrostro-Nava, M. T.; Michel-López, C. Y.; Bermúdez-Guzmán, M. J.; Guzmán-González, S.; Orozco-Santos, M.; Velázquez-Monreal, J. J.; Buenrostro-Nava, M. T. y Michel-López, C. Y. 2016. Optimización de un protocolo para aislamiento de DNA de hojas de Saccharum officinarum. Revista Mexicana de Ciencias Agrícolas. 7(4):897-910.
Bermúdez-Guzmán, M. J.; Guzmán-Rodríguez, L. F.; García-Mariscal, K. P.; Palmeros-Suárez, P. A.; Orozco-Santos, M.; Bermúdez-Guzmán, M. J.; Guzmán-Rodríguez, L. F.; García-Mariscal, K. P.; Palmeros-Suárez, P. A. y Orozco-Santos, M. 2017. Identificación de híbridos de Citrus aurantifolia×Citrus limon utilizando marcadores de secuencias simples repetidas (SSR). Revista Mexicana de Ciencias Agrícolas. 8(6):1397-1408. https://doi.org/10.29312/remexca.v8i6.309.
Carrillo-Medrano, S. H.; Gutierrez-Espinosa, M. A.; Robles-González, M. M.; Cruz-Izquierdo, S.; Carrillo-Medrano, S. H.; Gutierrez-Espinosa, M. A.; Robles-González, M. M. y Cruz-Izquierdo, S. 2018. Identificación de híbridos de limón mexicano mediante marcadores moleculares SSR. Revista Mexicana de Ciencias Agrícolas. 9(1):11-23. https://doi.org/10.29312/remexca.v9i1.844 https://doi.org/10.29312/remexca.v9i1.844.
Gallego-Colonia, J. S.; Enríquez-Valencia, A. L.; Caicedo-Arana, Á.; Posso-Terranova, A. M. y Muñoz-Florez, J. E. 2017. Diversidad genética en patrones de cítricos mediante microsatélites amplificados al azar (RAMs). Biotecnología en el Sector Agropecuario y Agroindustrial. 15(1):85-94. https://doi.org/10.18684/BSAA(15)85-94.
Graper, A. L.; Noyszewski, A. K.; Anderson, N. O. and Smith, A. G. 2021. Variability in ITS1 and ITS2 sequences of historic herbaria and extant (fresh) Phalaris species (Poaceae). BMC Plant Biology. 21(1):1-16. https://doi.org/10.1186/s12870-021-03284-z.
Hynniewta, M.; Malik, S. K. and Rao, S. R. 2014. Genetic diversity and phylogenetic analysis of Citrus (L.) from north-east India as revealed by meiosis, and molecular analysis of internal transcribed spacer region of rDNA. Meta Gene. 2:237-251. https://doi.org/10.1016/j.mgene.2014.01.008.
Ito, T. M.; Polido, P. B.; Rampim, M. C.; Kaschuk, G. and Souza, S. G. H. 2014. Genome-wide identification and phylogenetic analysis of the AP2/ERF gene superfamily in sweet orange (Citrus sinensis). Genetics and Molecular Research: GMR. 13(3):7839-7851. https://doi.org/10.4238/2014.September.26.22.
Jin, S. B.; Lee, W. J.; Park, J. H.; Park, S. M.; Lee, D. H. and Yun, S. H. 2018. A phylogenic analysis of citrus cultivars native to jeju using chloroplast dna trnl-trnf and internal transcribed spacer region sequences. Horticultural Science and Technology. 36(4):585-597. https://doi.org/10.12972/kjhst.20180059.
Kyndt, T.; Dung, T. N.; Goetghebeur, P.; Toan, H. T. and Gheysen, G. 2010. Analysis of ITS of the rDNA to infer phylogenetic relationships among Vietnamese Citrus accessions. Genetic Resources and Crop Evolution. 57(2):183-192. https://doi.org/10.1007/s10722-009-9460-0.
Letsiou, S.; Madesis, P.; Vasdekis, E.; Montemurro, C.; Grigoriou, M. E.; Skavdis, G.; Moussis, V.; Koutelidakis, A. E. and Tzakos, A. G. 2024. DNA Barcoding as a Plant Identification Method. Applied Sciences. 14(4):1-12. https://doi.org/10.3390/app14041415.
Li, X.; Xie, R.; Lu, Z. and Zhou, Z. 2010. The origin of cultivated citrus as inferred from internal transcribed spacer and chloroplast DNA sequence and amplified fragment length polymorphism fingerprints. Journal of the American Society for Horticultural Science. 135(4):341-350. https://doi.org/10.21273/JASHS.135.4.341.
Liu, M.; Wang, K.; Chen, B.; Cai, Y; Li, C.; Yang, W.; Wei, M. and Zheng, G. 2021. Intraspecific DNA barcoding and variation analysis for Citri reticulatae pericarpium of Citrus reticulata ‘Chachi’. Evidence-based Complementary and Alternative Medicine. eCAM, 2021, 2609935: 1-7 pp. https://doi.org/10.1155/2021/2609935.
Mahjbi, A.; Oueslati, A.; Baraket, G.; Salhi-Hannachi, A. and Zehdi-Azouzi, S. 2016. Assessment of genetic diversity of Tunisian orange, Citrus sinensis (L.) Osbeck using microsatellite (SSR) markers. Genetics and Molecular Research. 15(2):1-12. https://doi.org/10.4238/gmr.15026564.
Morgante, M. and Olivieri, A. M. 1993. PCR-amplified microsatellites as markers in plant genetics. The plant journal: for cell and molecular biology. 3(1):175-182. https://doi.org/10.1046/j.1365-313X.1993.t01-9-00999.x.
Penjor, T.; Yamamoto, M.; Uehara, M.; Ide, M.; Matsumoto, N.; Matsumoto, R. and Nagano, Y. 2013. Phylogenetic relationships of citrus and its relatives based on matk gene sequences. Plos One. 8(4):e62574. https://doi.org/10.1371/journal.pone.0062574.
Sankar, T. G.; Gopi, V.; Deepa, B. and Gopal, K. 2014. Genetic diversity analysis of sweet orange (Citrus sinensis Osbeck.) varieties/clones through RAPD markers. Int. J. Curr. Microbiol. App. Sci. 3(4):75-84.
Seminara, S.; Bennici, S.; Guardo, M. D.; Caruso, M.; Gentile, A.; Malfa, S. and Distefano, G. 2023. Sweet orange: evolution, characterization, varieties and breeding perspectives. Agriculture. 13(2):1-24. https://doi.org/10.3390/agriculture13020264.
Shahnazari, N.; Noormohammadi, Z.; Sheidai, M. and Koohdar, F. 2022. A new insight on genetic diversity of sweet oranges: CAPs-SSR and SSR markers. Journal of Genetic Engineering & Biotechnology. 20(105):1-8. https://doi.org/10.1186/s43141-022-00393-6.
Sharafi, A. A.; Abkenar, A. A. and Sharafi, A. 2017. Molecular genetic diversity assessment of Citrus species grown in Iran revealed by SSR, ISSR and CAPS molecular markers. Journal of Science and Research. 2(8):22-27. https://doi.org/10.26910/issn.2528-8083vol2iss8.2017pp22-27.
Tuwo, M.; Kuswinanti, T.; Nasruddin, A. and Tambaru, E. 2023. Estimating the genetic diversity of oranges Citrus spp. In: South Sulawesi, Indonesia, Using RAPD Markers. Scientifica. 6676038:1-12. https://doi.org/10.1155/2023/6676038.
Viglietti, G.; Galla, G.; Porceddu, A.; Barcaccia, G.; Curk, F.; Luro, F. and Scarpa, G. M. 2019. Karyological analysis and DNA barcoding of pompia citron: a first step toward the identification of its relatives. Plants. 8(4):1-16. https://doi.org/10.3390/plants8040083.
Wenger, A. M.; Peluso, P.; Rowell, W. J.; Chang, P. C.; Hall, R. J.; Concepcion, G. T.; Ebler, J.; Fungtammasan, A.; Kolesnikov, A.; Olson, N. D.; Töpfer, A.; Chin, C. S.; Alonge, M.; Mahmoud, M.; Qian, Y.; Phillippy, A. M.; Schatz, M. C.; Myers, G.; Pristo, M. A. and Hunkapiller, M. W. 2019. Highly-accurate long-read sequencing improves variant detection and assembly of a human genome. Preprint, bioRxiv, 519025. https://doi.org/10.1101/519025.
White, T.; Bruns, T.; Lee, S.; Taylor, J.; Innis, M.; Gelfand, D. and Sninsky, J. J. 1990. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. In: Innis, M. A.; Gelfand, D. H.; Sninsky, J. J. and White, T. J. (eds.). PCR Protocols: A Guide to Methods and Applications. Academic Press. San Diego, California, USA. 315-322 pp. https://doi.org/10.1016/b978-0-12-372180-8.50042-1.
Williams, J. G.; Kubelik, A. R.; Livak, K. J.; Rafalski, J. A. and Tingey, S. V. 1990. DNA polymorphisms amplified by arbitrary primers are useful as genetic markers. Nucleic Acids Research. 18(22):6531-6535. https://doi.org/10.1093/nar/18.22.6531.
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