Phenotypic and Nodule Microbial Diversity among Crimson Clover (Trifolium incarnatum L.) Accessions

Crimson clover (Trifolium incarnatum L.) is the most common legume cover crop in the United States. Previous research found limited genetic variation for crimson clover within the National Plant Germplasm System (NPGS) collection. The aim of this study was to assess the phenotypic and nodule microbial diversity within the NPGS crimson clover collection, focusing on traits important for cover crop performance. Experiments were conducted at the Beltsville Agricultural Research Center (Maryland, USA) across three growing seasons (2012–2013, 2013–2014, 2014–2015) to evaluate 37 crimson clover accessions for six phenotypic traits: fall emergence, winter survival, flowering time, biomass per plant, nitrogen (N) content in aboveground biomass, and proportion of plant N from biological nitrogen fixation (BNF). Accession effect was significant across all six traits. Fall emergence of plant introductions (PIs) ranged from 16.0% to 70.5%, winter survival ranged from 52.8% to 82.0%, and growing degree days (GDD) to 25% maturity ranged from 1470 GDD to 1910 GDD. Biomass per plant ranged from 1.52 to 6.51 g, N content ranged from 1.87% to 2.24%, and proportion of plant N from BNF ranged from 50.2% to 85.6%. Accessions showed particularly clear differences for fall emergence and flowering time, indicating greater diversity and potential for selection in cover crop breeding programs. Fall emergence and winter survival were positively correlated, and both were negatively correlated with biomass per plant and plant N from BNF. A few promising lines performed well across multiple key traits, and are of particular interest as parents in future breeding efforts, including PIs 369045, 418900, 561943, 561944, and 655006. In 2014–2015, accessions were also assessed for nodule microbiome diversity, and 11 genera were identified across the sampled nodules. There was large variation among accessions in terms of species diversity, but this diversity was not associated with observed plant traits, and the functional implications of nodule microbiome diversity remain unclear.