Abstract
Elymus sibiricus L. is an important cold-season grass with excellent cold and drought tolerance, good palatability, and nutrition. Flowering time is a key trait that affects forage and seed yield. Development of EST-SSR (expressed sequence tag simple sequence repeat) markers based on flowering genes contributes to the improvement of flowering traits. In the study, we detected 155 candidate genes related to flowering traits from 10,591 unigenes via transcriptome sequencing in early- and late-flowering genotypes. These candidate genes were mainly involved in the photoperiodic pathway, vernalization pathway, central integrator, and gibberellin pathway. A total of 125 candidate gene-based EST-SSRs were developed. Further, 15 polymorphic EST-SSRs closely associated to 13 candidate genes were used for genetic diversity and population structure analysis among 20 E. sibiricus accessions, including two contrasting panels (early-flowering and late-flowering). Among them, primer 28366, designed from heading date 3a (HD3a), effectively distinguished early- and late-flowering genotypes using a specifically amplified band of 175 bp. The polymorphic information content (PIC) value ranged from 0.12 to 0.48, with an average of 0.25. The unweighted pair group method analysis (UPGMA) cluster and structure analysis showed that the 20 E. sibiricus genotypes with similar flowering times tended to group together. These newly developed EST-SSR markers have the potential to be used for molecular markers assisted selection and germplasm evaluation of flowering traits in E. sibiricus.
Highlights
Elymus sibiricus L. is a perennial, predominantly self-pollinating and allotetraploid (2n = 4x = 28)forage grass of the genus Elymus
To better understand the mechanism for flowering in E. sibiricus, we developed novel EST-SSR
E. sibiricus, 18 cDNA libraries were constructed from leaves collected from early flowering genotypes and genotypes at booting stage,stage, heading stage, and flowering stage
Summary
Elymus sibiricus L. is a perennial, predominantly self-pollinating and allotetraploid (2n = 4x = 28). It is widely distributed in high altitude regions of western and northern China, especially on the Qinghai-Tibetan Plateau [1]. E. sibiricus is an important cold-season grass with excellent cold and drought tolerance, good palatability, and forage quality [2]. Research on breeding and improvement for this species is limited. The quantity of E. sibiricus varieties cannot meet the growing demands of ecological restoration and animal husbandry in China [3]. In perennial forage grasses, flowering indicates the transition from vegetative growth to reproductive growth, which affects characters of economic importance such as forage quality and seed yield [4]
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