Abstract
Evergreen azaleas are among the most important ornamental shrubs in China. Today, there are probably over 300 cultivars preserved in different nurseries, but with little information available on the cultivar itself or relationships between cultivars. Amplified fragment length polymorphism (AFLP) markers were employed to determine the genetic relationships between evergreen azalea cultivars in China. One hundred and thirty genotypes collected from gardens and nurseries, including cultivars classified in the groups East, West, Hairy, and Summer, unknown cultivars, and close species, were analyzed using three primer pairs. A total of 408 polymorphic fragments were generated by AFLP reactions with an average of 136 fragments per primer pair. The average values of expected heterozygosity and Shannon’s information index were 0.3395 and 0.5153, respectively. Genetic similarities were generated based on Dice coefficients, used to construct a neighbor joining tree, and bootstrapped for 100 replicates in Treecon V1.3b. Principal coordinate analysis (PCO) was performed based on Dice distances using NTSYS-pc software. The AFLP technique was useful for analyzing genetic diversity in evergreen azaleas. Cluster analysis revealed that cultivars in the West and Summer groups were quite distinct from other groups in the four-group classification system and that the East and Hairy groups should be redefined.
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References
Bonin, A., Ehrich, D., Manel, S., 2007. Statistical analysis of amplified fragment length polymorphism data: a toolbox for molecular ecologists and evolutionists. Mol. Ecol., 16(18):3737–3758. [doi:10.1111/j.1365-294X.2007.03435.x]
Chamberlain, D.F., Rae, S.J., 1990. A revision of Rhododendron. IV Subgenus Tsutsusi. Edinb. J. Bot., 47(2):89–200. [doi:10.1017/S096042860000319X]
de Riek, J., Dendauw, J., Mertens, M., de Loose, M., Heursel, J., van Blockstaele, E., 1999. Validation of criteria for the selection of AFLP markers to assess the genetic variation of a breeders’ collection of evergreen azaleas. Theor. Appl. Gen., 99(7–8):1155–1165. [doi:10.1007/s001220051320]
Dendauw, J., de Rick, J., de Loose, M., van Bockstaele, E., 2002. Identification of 33 Chinese Rhododendron species using MATK sequences and AFLP data. Proc. Twent. Int. Eucarp. Symp.-Sect. Orn., 572:169–177.
Escaravage, N., Questiau, S., Pornon, A., Doche, B., Taberlet, P., 1998. Clonal diversity in a Rhododendron ferrugineum L. (Ericaceae) population inferred from AFLP markers. Mol. Ecol., 7(8):975–982. [doi:10.1046/j.1365-294x.1998.00415.x]
Huang, M.R., Qiang, H.L., 1984. Rhododendron. China Forestry Publishing House, Beijing, China (in Chinese).
Iqbal, M.J., Paden, D.W., Rayburn, A.L., 1995. Assessment of genetic-relationships among Rhododendron species, varieties and hybrids by RAPD analysis. Sci. Hort., 63(3–4):215–223. [doi:10.1016/0304-4238(95)00808-7]
Kobayashi, N., Takeuchi, R., Handa, T., Takayanagai, K., 1995. Cultivar identification of evergreen azalea with RAPD method. J. Jpn. Soc. Hort. Sci., 64(3):611–616. [doi:10.2503/jjshs.64.611]
Kobayashi, N., Handa, T., Takayanagi, K., Arisumi, K., 2003. Clarification of origin in azalea cultivars by PCR-RFLP analysis of chloroplast DNA. J. Jpn. Soc. Agric. Technol. Man., 10(2):143–147 (in Japanese).
Koopman, W.J.M., Wissemann, V., de Cock, K., van Huylenbroeck, J., de Riek, J., Sabatlno, G.J.H., Visser, D., Vosman, B., Ritz, C.M., Maes, B., et al., 2008. AFLP markers as a tool to reconstruct complex relationships: a case study in rosa (Rosaceae). Am. J. Bot., 95(3):353–366. [doi:10.3732/ajb.95.3.353]
Kurashige, Y., Milne, M., Kobayashi, N., Handa, T., Takayanagi, K., Yukawa, T., 1998. Investigation of sectional relationships in the genus Rhododendron (Ericaceae) based on MATK sequences. J. Jpn. Bot., 73(2):143–154 (in Japanese).
Kurashige, Y., Etoh, J.I., Handa, T., Takayanagi, K., Yukawa, T., 2001. Sectional relationships in the genus Rhododendron (Ericaceae): evidence from MATK and TRNK intron sequences. Plant Syst. Evol., 228(1–2):1–14. [doi:10.1007/s006060170033]
Meudt, H.M., Clarke, A.C., 2007. Almost forgotten or latest practice? AFLP applications, analyses and advances. Trends Plant Sci., 12(3):106–117. [doi:10.1016/j.tplants.2007.02.001]
Meudt, H.M., Bayly, M.J., 2008. Phylogeographic patterns in the Australasian genus Chionohebe (Veronica s.l., Plantaginaceae) based on AFLP and chloroplast DNA sequences. Mol. Phyl. Evol., 47(1):319–338. [doi:10.1016/j.ympev.2007.12.019]
Nei, M., Li, W.H., 1979. Mathematical-model for studying genetic-variation in terms of restriction endonucleases. PNAS, 76(10):5269–5273. [doi:10.1073/pnas.76.10.5269]
Rohlf, F., 1998. NTSYS-PC, Numerical Taxonomy and Multivariate Analysis, Version 2.0. Exeter Software. Setauket, New York.
Scariot, V., de Keyser, E., Handa, T., de Riek, J., 2007a. Comparative study of the discriminating capacity and effectiveness of AFLP, STMS and EST markers in assessing genetic relationships among evergreen azaleas. Plant Breed., 126(2):207–212. [doi:10.1111/j.1439-0523.2007.01326.x]
Scariot, V., Handa, T., de Riek, J., 2007b. A contribution to the classification of evergreen azalea cultivars located in the lake maggiore area (Italy) by means of AFLP markers. Euphytjca, 158(1–2):47–66. [doi:10.1007/s10681-007-9425-3]
Shen, Y.C., 2004. The Illustrated Encyclopedia of the World Famous Azaleas. China Architecture & Building Press, Beijing, China (in Chinese).
Tsai, C.C., Huang, S.C., Chen, C.H., Tseng, Y.H., Huang, P.L., Tsai, S.H., Chou, C.H., 2003. Genetic relationships of Rhododendron (Ericaceae) in Taiwan based on the sequence of the internal transcribed spacer of ribosomal DNA. J. Hort. Sci. Biotechnol., 78(2):234–240.
van de Peer, Y., de Wachter, R., 1994. Treecon for windows-a software package for the construction and drawing of evolutionary trees for the microsoft windows environment. Comp. Appl. Biosci., 10(5):569–570. [doi:10.1093/bioinformatics/10.5.569]
Vos, P., Hogers, R., Bleeker, M., Reijans, M., Vandelee, T., Hornes, M., Frijters, A., Pot, J., Peleman, J., Kuiper, M., et al., 1995. AFLP—a new technique for DNA-fingerprinting. Nucleic Acids Res., 23(21):4407–4414. [doi:10.1093/nar/23.21.4407]
Yeh, F.C., Boyle, T.J.B., 1997. Population genetic analysis of co-dominant and dominant markers and quantitative traits. Belg. J. Bot., 129:157.
Zhao, J.J., Wang, X.W., Deng, B., Lou, P., Wu, J., Sun, R.F., Xu, Z.Y., Vromans, J., Koornneef, M., Bonnema, G., 2005. Genetic relationships within Brassica rapa as inferred from AFLP fingerprints. Theor. Appl. Gen., 110(7):1301–1314.
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Project (No. 2012C12909-7) supported by the Science and Technology Major Project of Zhejiang Province, China
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Zhou, H., Liao, J., Xia, Yp. et al. Determination of genetic relationships between evergreen azalea cultivars in China using AFLP markers. J. Zhejiang Univ. Sci. B 14, 299–308 (2013). https://doi.org/10.1631/jzus.B1200094
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DOI: https://doi.org/10.1631/jzus.B1200094
Key words
- Rhododendron spp.
- Tsutsusi
- Fluorescent label
- Dice coefficient
- Principal coordinate analysis (PCO)
- Neighbor joining tree