Cadmium soil contamination alters plant-pollinator interactions.

Soil heavy metal contamination is often an unintended byproduct of historic land-use. This contamination can negatively impact resident plants and their interactions with other organisms. Plant fitness in contaminated landscapes depends not only on plant growth, but also on the maintenance of interactions with pollinators. Cadmium (Cd) is a heavy metal that is commonly found in agricultural, urban, and industrial ecosystems as a legacy of historic land-use. It is a prioritized pollutant in soils because of its wide distribution and strong biotoxicity. To understand how Cd influences plant growth and pollinator interactions, we grew sunflowers in media with three different Cd concentrations to represent the range of Cd contamination faced by sunflowers growing on land recovering from past land-use. We measured Cd contamination effects on sunflower morphology and pollinator foraging behavior, specifically the number of visits and visit duration. We then measured seed number and weight to determine if contamination directly or indirectly, as mediated by pollinators, altered plant fitness. Plant height was negatively correlated with Cd concentration, but contamination alone (in the absence of pollinators) did not affect sunflower reproduction. Bumble bees visited sunflowers grown in Exceeding Threshold Cd concentrations less often and for shorter time compared to visits to Below Threshold Cd sunflowers, but honey bees and sweat bees showed similar foraging behavior across Cd contamination treatment levels. Sunflower seed set was positively correlated with the total number of pollinator visits, and sunflowers grown in Exceeding Threshold Cd soil had marginally lower seed set compared to those grown in Below Threshold Cd soil. Our results suggest that at Exceeding Threshold Cd contamination levels plant-pollinator interactions are negatively affected with consequences for plant fitness.

Evolutionary analysis of the OSCA gene family in sunflower (Helianthus annuus L) and expression analysis under NaCl stress.

Hyperosmolality-gated calcium-permeable channels (OSCA) are Ca2 + nonselective cation channels that contain the calcium-dependent DUF221 domain, which plays an important role in plant response to stress and growth. However, the OSCA gene has not been fully identified and analyzed in sunflowers. In this study, we comprehensively analyzed the number, structure, collinearity, and phylogeny of the OSCA gene family in the sunflower, six Compositae species (Arctium lappa, Chrysanthemum morifolium, Cichorium endivia, Cichorium intybus, Lactuca sativa var. Angustata, and Carthamus tinctorius), and six other plants (soybean, Arabidopsis thaliana, rice, grape, and maize). The expression of the sunflower OSCA gene in nine different tissues, six different hormones, and NaCl stress conditions were analyzed based on transcriptome data and qRT-PCR. A total of 15 OSCA proteins, distributed on 10 chromosomes, were identified in the sunflower, and all of them were located in the endoplasmic reticulum. Using the phylogenetic tree, collinearity, gene structure, and motif analysis of the six Compositae species and six other plants, we found that the sunflower OSCA protein had only three subfamilies and lacked the Group 4 subfamily, which is conserved in the evolution of Compositae and subject to purification selection. The OSCA gene structure and motif analysis of the sunflower and six Compositae showed that there was a positive correlation between the number of motifs of most genes and the length of the gene, different subfamilies had different motifs, and the Group 4 subfamily had the smallest number of genes and the simplest gene structure. RNA-seq and qRT-PCR analysis showed that the expression levels of most OSCA genes in the sunflower changed to varying degrees under salt stress, and HaOSCA2.6 and HaOSCA3.1 were the most important in the sunflower's response to salt stress. The coexpression network of the sunflower genes under salt stress was constructed based on weighted gene co-expression network analysis (WGCNA). In conclusion, our findings suggest that the OSCA gene family is conserved during the sunflower's evolution and plays an important role in salt tolerance. These results will deepen our understanding of the evolutionary relationship of the sunflower OSCA gene family and provide a basis for their functional studies under salt stress.

Monotone Circuit Lower Bounds from Robust Sunflowers.

Robust sunflowers are a generalization of combinatorial sunflowers that have applications in monotone circuit complexity Rossman (SIAM J. Comput. 43:256-279, 2014), DNF sparsification Gopalan et al. (Comput. Complex. 22:275-310 2013), randomness extractors Li et al. (In: APPROX-RANDOM, LIPIcs 116:51:1-13, 2018), and recent advances on the Erdos-Rado sunflower conjecture Alweiss et al. (In: Proceedings of the 52nd Annual ACM SIGACT Symposium on Theory of Computing, STOC. Association for Computing Machinery, New York, NY, USA, 2020) Lovett et al. (From dnf compression to sunflower theorems via regularity, 2019) Rao (Discrete Anal. 8,2020). The recent breakthrough of Alweiss, Lovett, Wu and Zhang Alweiss et al. (In: Proceedings of the 52nd Annual ACM SIGACT Symposium on Theory of Computing, STOC. Association for Computing Machinery, New York, NY, USA, 2020) gives an improved bound on the maximum size of a w-set system that excludes a robust sunflower. In this paper, we use this result to obtain an exp ( n 1 / 2 - o ( 1 ) ) lower bound on the monotone circuit size of an explicit n-variate monotone function, improving the previous best known exp ( n 1 / 3 - o ( 1 ) ) due to Andreev (Algebra and Logic, 26:1-18, 1987) and Harnik and Raz (In: Proceedings of the Thirty-Second Annual ACM Symposium on Theory of Computing, ACM, New York, 2000). We also show an exp ( Ω ( n ) ) lower bound on the monotone arithmetic circuit size of a related polynomial via a very simple proof. Finally, we introduce a notion of robust clique-sunflowers and use this to prove an n Ω ( k ) lower bound on the monotone circuit size of the CLIQUE function for all k ⩽ n 1 / 3 - o ( 1 ) , strengthening the bound of Alon and Boppana (Combinatorica, 7:1-22, 1987).

Monitoring Alternaria toxins in Italian food to support upcoming regulation.

The collection of occurrence data on Alternaria toxins in food and feed across the European countries is required since 2012 by the European Commission, endorsing the relevant scientific opinion by the EFSA CONTAM Panel. Within this framework, occurrence data for Alternaria toxins (Alternariol, Alternariol monomethyl ether, Tenuazonic acid, Tentoxin, and Altenuene) in 97 samples of cereal foods, tomato products, and sunflower seeds have been provided as requested by the Italian national monitoring programme (years 2017-2020). To this purpose, an LC-MS/MS method was set up and validated, obtaining fit for purpose sensitivity, recoveries (70-120%), repeatability (≤20%) and within laboratory reproducibility (≤26%). Occurrence data showed that oilseeds were the most contaminated food group with levels of Tenuazonic acid up to 16752 µg/kg and Tentoxin up to 570 µg/kg, whereas for the other mycotoxin/commodities combinations, the percentage of left censored data (below the limit of quantification) ranged from 74 to 100%.

The Insights into Mitochondrial Genomes of Sunflowers.

The significant difference in the mtDNA size and structure with simultaneous slow evolving genes makes the mitochondrial genome paradoxical among all three DNA carriers in the plant cell. Such features make mitochondrial genome investigations of particular interest. The genus Helianthus is a diverse taxonomic group, including at least two economically valuable species-common sunflower (H. annuus) and Jerusalem artichoke (H. tuberosus). The successful investigation of the sunflower nuclear genome provided insights into some genomics aspects and significantly intensified sunflower genetic studies. However, the investigations of organelles' genetic information in Helianthus, especially devoted to mitochondrial genomics, are presented by limited studies. Using NGS sequencing, we assembled the complete mitochondrial genomes for H. occidentalis (281,175 bp) and H. tuberosus (281,287 bp) in the current investigation. Besides the master circle chromosome, in the case of H. tuberosus, the 1361 bp circular plasmid was identified. The mitochondrial gene content was found to be identical for both sunflower species, counting 32 protein-coding genes, 3 rRNA, 23 tRNA genes, and 18 ORFs. The comparative analysis between perennial sunflowers revealed common and polymorphic SSR and SNPs. Comparison of perennial sunflowers with H. annuus allowed us to establish similar rearrangements in mitogenomes, which have possibly been inherited from a common ancestor after the divergence of annual and perennial sunflower species. It is notable that H. occidentalis and H. tuberosus mitogenomes are much more similar to H. strumosus than H. grosseserratus.

The Investigation of Perennial Sunflower Species (Helianthus L.) Mitochondrial Genomes.

The genus Helianthus is a diverse taxonomic group with approximately 50 species. Most sunflower genomic investigations are devoted to economically valuable species, e.g., H. annuus, while other Helianthus species, especially perennial, are predominantly a blind spot. In the current study, we have assembled the complete mitogenomes of two perennial species: H. grosseserratus (273,543 bp) and H. strumosus (281,055 bp). We analyzed their sequences and gene profiles in comparison to the available complete mitogenomes of H. annuus. Except for sdh4 and trnA-UGC, both perennial sunflower species had the same gene content and almost identical protein-coding sequences when compared with each other and with annual sunflowers (H. annuus). Common mitochondrial open reading frames (ORFs) (orf117, orf139, and orf334) in sunflowers and unique ORFs for H. grosseserratus (orf633) and H. strumosus (orf126, orf184, orf207) were identified. The maintenance of plastid-derived coding sequences in the mitogenomes of both annual and perennial sunflowers and the low frequency of nonsynonymous mutations point at an extremely low variability of mitochondrial DNA (mtDNA) coding sequences in the Helianthus genus.

[Effect of Fertilizers on Cadmium Uptake and Accumulation by Sunflowers].

A pot experiment was conducted to assess the phytoextraction potential for cadmium (Cd) of three types of sunflowers (edible, ornamental, and oil sunflowers) and the effects of the application of N, NP, and NPK fertilizers on Cd uptake of the three plants using Cd-contaminated soils collected from northern China. The results showed that fertilization could significantly increase the biomass, aboveground Cd concentrations and accumulation of ornamental and oil sunflowers, and the effect of NPK fertilizer was significantly better than those of other treatments. Cd concentrations of the aboveground parts of edible, ornamental, and oil sunflowers were 6.89, 8.92, 6.97 mg·kg-1, respectively. Fertilization treatment significantly improved the transport ability of Cd of the three types of sunflowers, and bioconcentration factors of edible, ornamental, and oil sunflowers were 2.63 (control) to 3.10 (NPK fertilizer), 2.80 (control) to 4.02 (NPK fertilizer), and 2.11 (control) to 3.14 (NPK fertilizer), respectively. The results further showed that the metal-enriched granules and cellular debris were the main enrichment sources in the subcellular fraction of the three types of sunflowers (more than 55%). In summary, sunflowers can be considered as plant material for remediation of Cd-contaminated soil. In addition, NPK fertilizers can effectively improve the efficiency of sunflowers.

Different leachate phytotreatment systems using sunflowers.

The use of energy crops in the treatment of wastewaters is of increasing interest, particularly in view of the widespread scarcity of water in many countries and the possibility of obtaining renewable fuels of vegetable origin. The aim of this study was to evaluate the feasibility of landfill leachate phytotreatment using sunflowers, particularly as seeds from this crop are suitable for use in biodiesel production. Two different irrigation systems were tested: vertical flow and horizontal subsurface flow, with or without effluent recirculation. Plants were grown in 130L rectangular tanks placed in a special climatic chamber. Leachate irrigated units were submitted to increasing nitrogen concentrations up to 372mgN/L. Leachate was successfully tested as an alternative fertilizer for plants and was not found to inhibit biomass development. The experiment revealed good removal efficiencies for COD (η>50%) up until flowering, while phosphorous removal invariably exceeded 60%. Nitrogen removal rates decreased over time in all experimental units, particularly in vertical flow tanks. In general, horizontal flow units showed the best performances in terms of contaminant removal capacity; the effluent recirculation procedure did not improve performance. Significant evapo-transpiration was observed, particularly in vertical flow units, promoting removal of up to 80% of the inlet irrigation volume.

Macroevolution of leaf defenses and secondary metabolites across the genus Helianthus.

Leaf defenses are widely recognized as key adaptations and drivers of plant evolution. Across environmentally diverse habitats, the macroevolution of leaf defenses can be predicted by the univariate trade-off model, which predicts that defenses are functionally redundant and thus trade off, and the resource availability hypothesis, which predicts that defense investment is determined by inherent growth rate and that higher defense will evolve in lower resource environments. Here, we examined the evolution of leaf physical and chemical defenses and secondary metabolites in relation to environmental characteristics and leaf economic strategy across 28 species of Helianthus (the sunflowers). Using a phylogenetic comparative approach, we found few evolutionary trade-offs among defenses and no evidence for defense syndromes. We also found that leaf defenses are strongly related to leaf economic strategy, with higher defense in more resource-conservative species, although there is little support for the evolution of higher defense in low-resource habitats. A wide variety of physical and chemical defenses predict resistance to different insect herbivores, fungal pathogens, and a parasitic plant, suggesting that most sunflower defenses are not redundant in function and that wild Helianthus represents a rich source of variation for the improvement of crop sunflower.