Fingerprint identification of white clover cultivars based on SSR molecular markers
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ORIGINAL ARTICLE
Fingerprint identification of white clover cultivars based on SSR molecular markers Sainan Ma1 · Chongyang Han1 · Jie Zhou1 · Ruchang Hu1 · Xing Jiang1 · Feifei Wu1 · Ke Tian1 · Gang Nie1 · Xinquan Zhang1 Received: 12 June 2020 / Accepted: 3 October 2020 © Springer Nature B.V. 2020
Abstract White clover (Trifolium repens L.) is an important perennial legume forage with high productivity and quality. To strengthen the basic research on the genetic characteristics, fingerprint identification and adaptability of white clover germplasm resources, Simple sequence repeat (SSR) molecular markers were applied to 10 white clover cultivars to assess the genetic diversity and related lines of white clover at the molecular level in order to lay a theoretical foundation for the selection of high-quality seeds and cultivars of white clover. A total of 120 different bands were amplified by 29 pairs of SSR primers with good polymorphism, of which 103 (89.5%) were polymorphic. Meanwhile, the PIC of each primer was 0.181–0.588, with an average of 0.329. Analysis of molecular variance revealed that 57% of the genetic variation occurred within cultivars and 43% occurred among cultivars. The results of cluster analysis and the principal coordinate analysis revealed that the parental relationships of the 10 cultivars, with the ‘Purple’ cultivar very distantly related to the other 9 cultivars and the closest parental relationship between ‘Ladino’ and ‘Sulky’. The fingerprints constructed by three representative primers (gtrs679, gtrs319, and gtrs678) have a strong identification ability. In summary, the SSR markers had good polymorphism and could be used for DNA fingerprint analysis of white clover cultivars. Keywords Trifolium repens L. · DNA fingerprinting · SSR · Genetic diversity
Introduction White clover (Trifolium repens L.) is one of the most widely planted perennial legume forages worldwide. It can not only produce high-quality forage but also be used as an ornamental plant with good ecological and economic value [1–3]. As an allotetraploid (2n = 4x = 32) species, it is has an obligate outbreeding habit and shows a high degree of Sainan Ma and Chongyang Han are the co-first authors. Electronic supplementary material The online version of this article (https://doi.org/10.1007/s11033-020-05893-7) contains supplementary material, which is available to authorized users. * Gang Nie [email protected] * Xinquan Zhang [email protected] 1
Department of Grassland Science, Animal Science and Technology College, Sichuan Agricultural University, Chengdu 611130, Sichuan, China
incompatibility, causing high genetic variation within populations [2]. Although white clover has an abundance of germplasm resources, many cultivars showed similar phenotypes due to the high natural rate of gene flow [4, 5], which limits the use of traditional variety identification methods based on morphological characteristics [6]. Therefore, the ability to reliably identify white clover cultivars is becoming increasingly important.
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