Assembly of high-quality genomes of the locoweed Oxytropis ochrocephala and its endophyte Alternaria oxytropis provides new evidence for their symbiotic relationship and swainsonine biosynthesis.

Locoweeds are perennial forbs poisonous to livestock and cause extreme losses to animal husbandry. Locoweed toxicity is attributed to the symbiotic endophytes in Alternaria sect. Undifilum, which produce a mycotoxin swainsonine (SW). We performed a de novo whole genome sequencing of the most common locoweed in China, Oxytropis ochrocephala (2n = 16), and assembled a high-quality, chromosome-level reference genome. Its genome size is 958.83 Mb with 930.94 Mb (97.09%) anchored and oriented onto eight chromosomes, and 31,700 protein-coding genes were annotated. Phylogenetic and collinearity analysis showed it is closely related to Medicago truncatula with a pair of large interchromosomal rearrangements, and both species underwent a whole-genome duplication event. We also derived the genome of A. oxytropis at 74.48 Mb with a contig N50 of 8.87 Mb and 10,657 protein-coding genes, and refined the genes of SW biosynthesis. Multiple Alternaria species containing the swnK gene were grouped into a single clade, but in other genera, swnK's homologues are diverse. Resequencing of 41 A. oxytropis strains revealed one SNP in the SWN cluster causing changes in SW concentration. Comparing the transcriptomes of symbiotic and nonsymbiotic interactions identified differentially expressed genes (DEGs) linked to defence and secondary metabolism in the host. Within the endophyte DEGs were linked to cell wall degradation, fatty acids and nitrogen metabolism. Symbiosis induced the upregulation of most of the SW biosynthetic genes. These two genomes and relevant sequencing data should provide valuable genetic resources for the study of the evolution, interaction, and SW biosynthesis in the symbiont.

Effects of Nitrogen Forms on the Growth and Nitrogen Accumulation in Buchloe dactyloides Seedlings.

Buffalograss [Buchloe dactyloides (Nutt.) Engelm.] has become the most widely cultivated warm-season turfgrass in northern China because of its low-maintenance requirements. Nitrogen (N) can be applied to plants in a range of formulations. However, preference of nitrogen uptake and the effects of N form on plant growth and nitrogen accumulation has not been established in buffalograss. In this study, we evaluated the effects of different inorganic nitrogen forms (NO3--N, NH4+-N, and NO3--N: NH4+-N = 1:1) on growth and nitrogen accumulation in buffalograss seedlings. Results showed that supply of three N forms significantly increased buffalograss seedlings growth, biomass, and N contents of all plant organs compared with the seedlings receiving free nitrogen. Plants achieved better growth performance when they received nitrate as the sole N source, which stimulated stolon growth and increased the biomass of ramets, spacers, and aboveground and total plant biomass, and also allocated more biomass to ramets and more N to spacers. Meanwhile, those plants supplied with the treatment +NH4NO3 displayed a significantly greater N content in the ramet, 15N abundance, and 15N accumulation amount in all organs. These data suggest NO3--N supplied either singly or in mixture increased vegetative propagation and thus facilitates buffalograss establishment. However, applications of ammonium caused detrimental effects on buffalograss seedlings growth, but +NO3- could alleviate NH4+-induced morphological disorders. Thus, recommendations to increase vegetative propagation and biomass accumulation in buffalograss seedlings should consider increasing NO3--N in a fertility program and avoiding applications of nitrogen as NH4+-N.

Biogeographic Patterns of Leaf Element Stoichiometry of Stelera chamaejasme L. in Degraded Grasslands on Inner Mongolia Plateau and Qinghai-Tibetan Plateau.

Plant leaf stoichiometry reflects its adaptation to the environment. Leaf stoichiometry variations across different environments have been extensively studied in grassland plants, but little is known about intraspecific leaf stoichiometry, especially for widely distributed species, such as Stellera chamaejasme L. We present the first study on the leaf stoichiometry of S. chamaejasme and evaluate its relationships with environmental variables. S. chamaejasme leaf and soil samples from 29 invaded sites in the two plateaus of distinct environments [the Inner Mongolian Plateau (IM) and Qinghai-Tibet Plateau (QT)] in Northern China were collected. Leaf C, N, P, and K and their stoichiometric ratios, and soil physicochemical properties were determined and compared with climate information from each sampling site. The results showed that mean leaf C, N, P, and K concentrations were 498.60, 19.95, 2.15, and 6.57 g kg-1; the average C:N, C:P, N:P, N:K and K:P ratios were 25.20, 245.57, 9.81, 3.13, and 3.21, respectively. The N:P:K-ratios in S. chamaejasme leaf might imply that its growth is restricted by K- or K+N. Moreover, the soil physicochemical properties in the S. chamaejasme-infested areas varied remarkably, and few significant correlations between S. chamaejasme leaf ecological stoichiometry and soil physicochemical properties were observed. These indicate the nutrient concentrations and stoichiometry of S. chamaejasme tend to be insensitive to variations in the soil nutrient availability, resulting in their broad distributions in China's grasslands. Besides, different homeostasis strength of the C, N, K, and their ratios in S. chamaejasme leaves across all sites were observed, which means S. chamaejasme could be more conservative in their use of nutrients improving their adaptation to diverse conditions. Moreover, the leaf C and N contents of S. chamaejasm were unaffected by any climate factors. However, the correlation between leaf P content and climate factors was significant only in IM, while the leaf K happened to be significant in QT. Besides, MAP or MAT contribution was stronger in the leaf elements than soil by using mixed effects models, which illustrated once more the relatively weak effect of the soil physicochemical properties on the leaf elements. Finally, partial least squares path modeling suggested that leaf P or K contents were affected by different mechanisms in QT and IM regions, suggesting that S. chamaejasme can adapt to changing environments by adjusting its relationships with the climate or soil factors to improve its survival opportunities in degraded grasslands.

Host-Species Variation and Environment Influence Endophyte Symbiosis and Mycotoxin Levels in Chinese Oxytropis Species.

Oxytropis plants are widely distributed in the grasslands in northern China. Some Oxytropis species have been reported to contain the mycotoxin swainsonine, an alkaloid which causes poisoning in livestock, referred to as locoism. Previous studies showed that endophytic fungi (Alternaria oxytropis) symbiotically associate with these Oxytropis species to produce swainsonine. However, the influence of variation within the Oxytropis genus on the fixation or loss of symbiosis and toxicity is poorly understood, as is the influence of environmental factors. Here we used a collection of 17 common Oxytropis species sampled in northern China to assess genetic diversity using genotyping by sequencing which was compared with the levels of the endophyte and swainsonine. Results showed that nine Oxytropis species have detectable A. oxytropis colonisation, and seven Oxytropis species contain sufficient swainsonine to be considered poisonous, whereas the rest may be non-toxic. Species variation rather than the genetic lineage was associated with the fixation or loss of endophyte and swainsonine production, which appears to have resulted from genetic drift. Genotype × Environment (G × E) effects were also found to influence endophyte and swainsonine levels amongst species of the Oxytropis genus. Our study will provide a better understanding about the evolutionary basis of A. oxytropis symbiosis and swainsonine biosynthesis in locoweeds.