Application of microbe-impregnated tannery solid waste biochar in soil enhances growth performance of sunflower.

Synergistic effect of biochar and microbes in soil enhances performance of plants. Hazardous tannery solid waste can be reduced by one-third in volume by conversion to biochar. A greenhouse trial was set up with soil having different doses of metal resistant microbe-impregnated biochar (MIBC) prepared from tannery solid waste. Consortia of autochthonous strains of Trichoderma and Bacillus were inoculated on BC and the behavior and fate of metals were evaluated for their bioavailability to sunflower. Sunflower was grown in pots for 80 days having six different amendments of tannery solid waste biochar (0-10% w/w) with and without Trichoderma and Bacillus consortia and its morphological and biochemical attributes as well as metal uptake were observed. The results illustrated that application of BC at 2% rate without inoculation increased the shoot length and dry biomass by 19.8% and 77.4%, respectively, while plant growth and performance were reduced at higher amendments of BC. However, application of MIBC with Trichoderma or/and Bacillus consortium significantly improved the plant attributes at all levels of amendment. The results indicated that MIBC having Trichoderma and Bacillus consortia at 10% rate increased shoot length and dry biomass by 65.3% and 516% compared to control without BC. Application of BC without inoculation reduced the uptake of Cu, Fe, and Ni and increased the mobilization of all other metals for uptake in sunflower. Mobilization and uptake of Cd, Cr, Cu, Ni, Pb, and Zn decreased with MIBC having Trichoderma and Bacillus consortia whereas that of Fe and Mg were noted. A considerable decrease in proline and total phenolic content was demonstrated by MIBC-grown sunflower. The data of metal fractionation in BC also supported the above findings. Therefore, MIBC can be used as a promising option for enhancing growth performance and ensuring the physiological safety of sunflower as an energy crop.

Comparative study of microbial structure and functional profile of sunflower rhizosphere grown in two fields.

BACKGROUND: Microbial communities inhabiting the rhizosphere play pivotal roles in determining plant health and yield. Manipulation of the rhizosphere microbial community is a promising means to enhance the productivity of economically viable and important agricultural crops such as sunflower (Helianthus annuus). This study was designed to gain insights into the taxonomic and functional structures of sunflower rhizosphere and bulk soil microbiome at two different locations (Sheila and Itsoseng) in South Africa. RESULTS: Microbial DNA extracted from the sunflower rhizosphere and bulk soils was subjected to next-generation sequencing using 16S amplicon sequencing technique. Firmicutes, Actnobacteria and Proteobacteria predominated sunflower rhizosphere soils. Firmicutes, Cyanobacteria, Deinococcus-Thermus and Fibrobacteres were positively influenced by Na+ and clay content, while Actinobacteria, Thaumarchaeota, Bacteroidetes, Planctomycetes, Aquificae and Chloroflexi were positively influenced by soil resistivity (Res) and Mg2+. The community-level physiological profiling (CLPP) analysis showed that the microbial communities in SHR and ITR used the amino acids tryptophan and malic acid efficiently. The metabolisms of these carbon substrates may be due to the dominant nature of some of the organisms, such as Actinobacteria in the soils. CONCLUSION: The CLPP measurements of soil from sunflower rhizosphere were different from those of the bulk soil and the degree of the variations were based on the type of carbon substrates and the soil microbial composition. This study has shown the presence of certain taxa of rhizobacteria in sunflower rhizosphere which were positively influenced by Na+ and Mg2+, and taxa obtained from SHR and ITR were able to effectively utilized tryptophan and malic acid. Many unclassified microbial groups were also discovered and it is therefore recommended that efforts should further be made to isolate, characterize and identify these unclassified microbial species, as it might be plausible to discover new microbial candidates that can further be harnessed for biotechnological purpose.

Methionine-induced regulation of growth, secondary metabolites and oxidative defense system in sunflower (Helianthus annuus L.) plants subjected to water deficit stress.

Optimum water availability at different growth stages is one the major prerequisites of best growth and yield production of plants. Exogenous application of plant growth regulators considered effective for normal functioning of plants under water-deficit conditions. A study was conducted to examine the influence of exogenously applied L-methionine on sunflower (Helianthus annuus L.) plants grown under water-deficit conditions. Twenty-five-day old seedlings of four sunflower cultivars, FH331, FH572, FH652 and FH623 were exposed to control (100% F.C.) and drought stress (60% F.C.) conditions. After 30-day of drought stress, L-methionine (Met; 20 mg/L) was applied as a foliar spray to control and drought stressed plants. Water deficit stress significantly reduced shoot fresh and dry weights shoot and root lengths, and chlorophyll a content in all four cultivars. While a significant increase was observed due to water deficiency in relative membrane permeability (RMP), malondialdehyde (MDA), total soluble proteins (TSP), total soluble sugars (TSS), ascorbic acid (AsA) and activity of peroxidase (POD). Although, exogenously applied Met was effective in decreasing RMP, MDA and H2O2 contents, it increased the shoot fresh weight, shoot length, chlorophyll a, chlorophyll a/b ratio, proline contents and the activities of SOD, POD and CAT enzymes in all four cultivars under water deficit stress. No change in AsA and total phenolics was observed due to foliar-applied Met under water stress conditions. Of all sunflower cultivars, cv. FH-572 was the highest and cv. FH-652 the lowest of all four cultivars in shoot fresh and dry weights as well as shoot length under drought stress conditions. Overall, foliar applied L-methionine was effective in improving the drought stress tolerance of sunflower plants that was found to be positively associated with Met induced improved growth attributes and reduced RMP, MDA and H2O2 contents under water deficit conditions.

Learning of a mimic odor combined with nectar nonsugar compounds enhances honeybee pollination of a commercial crop.

The increasing demand on pollination services leads food industry to consider new strategies for management of pollinators to improve their efficiency in agroecosystems. Recently, it was demonstrated that feeding beehives food scented with an odorant mixture mimicking the floral scent of a crop (sunflower mimic, SM) enhanced foraging activity and improved recruitment to the target inflorescences, which led to higher density of bees on the crop and significantly increased yields. Besides, the oral administration of nonsugar compounds (NSC) naturally found in nectars (caffeine and arginine) improved short and long-term olfactory memory retention in conditioned bees under laboratory conditions. To test the effect of offering of SM-scented food supplemented with NSC on honeybees pollinating sunflower for hybrid seed production, in a commercial plantation we fed colonies SM-scented food (control), and SM-scented food supplemented with either caffeine, arginine, or a mixture of both, in field realistic concentrations. Their foraging activity was assessed at the hive and on the crop up to 90 h after treatment, and sunflower yield was estimated prior to harvest. Our field results show that SM + Mix-treated colonies exhibited the highest incoming rates and densities on the crop. Additionally, overall seed mass was significantly higher by 20% on inflorescences close to these colonies than control colonies. Such results suggest that combined NSC potentiate olfactory learning of a mimic floral odor inside the hive, promoting faster colony-level foraging responses and increasing crop production.

Conservation tillage improves productivity of sunflower (Helianthus annuus L.) under reduced irrigation on sandy loam soil.

Sunflower production is significantly lower in arid and semi-arid regions due to various crop management problem. Conservation of tillage provides the most excellent opportunity to reduce degradation of soil reserves and increase soil productivity. The main objective of this study was to investigate the combined effects of conservation tillage and drought stress on growth and productivity of different sunflower hybrids. Experimental treatments included two sunflower hybrids ('NK-Senji' and 'S-278'), two drought stress treatments (i.e., well-watered and drought stress at flowering and grain filling stages) and three tillage practices (i.e., conservation, minimum and deep tillage). The results indicated that morphological and physiological parameters, and yield-related traits were significantly (P≤0.05) affected by all individual factors; however, their interactive effects were non-significant. Among sunflower hybrids, 'NK-Senji' performed better for morphological, physiological, and yield-related traits than 'S-278'. Similarly, conservation tillage observed better traits compared to the rest of the tillage practices included in the study. Nonetheless, conservation tillage improved growth and yield-related traits of hybrid 'NK-Senji' under drought stress. Hence, it is concluded that conservation tillage can improve the productivity of sunflower under low moisture availability. Therefore, conservation tillage could be suggested in the areas of lower water ability to improve sunflower production. Nonetheless, sunflower hybrids or varieties need thorough testing for their adaptability to conservation tillage and low moisture availability before making recommendations.

Potassium fertilization improves growth, yield and seed quality of sunflower (Helianthus annuus L.) under drought stress at different growth stages.

Water scarcity is a major concern for sunflower production in the semi-arid and arid regions of the world. Potassium (K) application has been found effective to alleviate the influence of drought stress; however, the impact of drought stress on seed quality of sunflower has not been reported frequently. Therefore, a field experiment was performed to determine the optimum K requirement for mitigating the adverse effects of water stress and improving growth and seed quality of spring-planted sunflower. Sunflower plants were exposed to water stress at different growth stages, i.e., Io = no stress (normal irrigation), I1 = pre-anthesisi stress (irrigation skipped at pre-anthesis stage), I2 = anthesis stress (irrigation skipped at anthesis stage) and I3 = post-anthesis stress (irrigation skipped at post-anthesis stage). Potassium was applied at four different rates, i.e., Ko = 0, K1 = 50, K2 = 100 and K3 = 150 kg ha-1. The results revealed that water stress at pre- and post-anthesis stages significantly reduced plant height, head diameter, number of achenes, oleic acid contents, and phosphorus (P) uptake. However, pre-anthesis stress improved linoleic acid contents. Treatment IoK3 (stress-free with 150 kg ha-1 K) was optimum combination for 1000-achene weight, biological and achene yields, oil contents, protein contents, and N and P uptake. Results indicated that a higher amount of K and irrigation resulted in higher yield, whereas yield and yield components decreased with early-stage water stress. Nevertheless, potassium application lowered the impacts of waters stress compared to no application. Keeping in view these results, it is recommended that sunflower must be supplied 150 kg ha-1 K in arid and semi-arid regions to achieve higher yield and better seed quality.

Signaling mechanisms and biochemical pathways regulating pollen-stigma interaction, seed development and seedling growth in sunflower under salt stress.

Sunflower (Helianthus annuus L.) is one of the major oilseed crops cultivated world over for its high-quality oil rich in linoleic acid. It also has established applications in pharmaceutical and biotechnological industries, mainly through recombinant production of unique oil body (OB) membrane proteins-oleosins, which are used for producing a wide variety of vaccines, food products, cosmetics and nutraceuticals. The present review provides a critical analysis of the progress made in advancing our knowledge in sunflower biology, ranging from mechanisms of pollen-stigma interaction, seed development, physiology of seed germination and seedling growth under salt stress, and finally understanding the signaling routes associated with various biochemical pathways regulating seedling growth. Role of nitric oxide (NO) triggered post-translational modifications (PTMs), discovered in the recent past, have paved way for future research directions leading to further understanding of sunflower developmental physiology. Novel protocols recently developed to monitor temporal and spatial distributions of various biochemicals involved in above-stated developmental events in sunflower, will go a long way for similar applications in plant biology in future.

Key factors involved in reduction of damage to sunflower by the European sunflower moth in China through late planting.

The European sunflower moth, Homoesoma nebulellum (Denis et Schiffermüller), emerged as a major new pest in Bayannur, China, in 2006. Insecticidal control with a single application is problematic because timing is critical, and multiple applications increase production and environmental costs. Management of H. nebulellum by planting date adjustment can be effective, but the optimal time window for late planting is unknown. Natural levels of H. nebulellum infestation were compared among sunflowers planted on five dates from April 25 to June 5 in two years, and the relationship between timing of adult abundance and flowering assessed. Delaying planting of sunflower from the traditional planting period of April 25 -May 5 to May 15 -June 5 significantly decreased damage by H. nebulellum. Seed infestation rate was 30-40 times higher, and number of larvae/head 75-100 times higher in the earliest two plantings than in the latest two. Within two years of implementing delayed planting in Bayannur city, infestation area decreased from 72% in 2006 to 1.5% in 2008, and production losses decreased from 4.5 ton/ha in 2006 to 0.36 ton/ha in 2008, a 97% decrease compared to 2006. Moreover, the infestation area caused by H. nebulellum was continuously controlled below 5.3% of the planting area since 2008. We found the overlap between the first two days of flowering and peak adult presence was the key factor influencing level of damage caused by H. nebulellum. Because the number of eggs laid in the first two days of flowering accounted for 68% of the total, and sunflower seed infestation rate was positively correlated with the number of trapped adults weighted by proportion of daily oviposition. Oviposition of the majority of eggs in the first two days of flowering suggests an evolutionary mechanism whereby females choose host plants most conducive to larval development, consistent with the preference-performance hypothesis.

Combination of arbuscular mycorrhizal fungi and phosphate solubilizing bacteria on growth and production of Helianthus tuberosus under field condition.

In this work, the effects of co-inoculation between an arbuscular mycorrhizal fungus (AMF) and a phosphate solubilizing bacteria (PSB) to promote the growth and production of sunchoke under field condition were investigated during 2016 and 2017. Four treatments were set up as follows: plants without inoculation, with AMF inoculation, with PSB inoculation and with co-inoculation of PSB and AMF. The results showed the presence of PSB and AMF colonization at the harvest stage in both years. This suggested the survival of PSB and successful AMF colonization throughout the experiments. According to correlation analysis, PSB positively affected AMF spore density and colonization rate. Also, both AMF and PSB positively correlated with growth and production of sunchoke. Co-inoculation could enhance various plant parameters. However, better results in 2016 were found in co-inoculation treatment, while AMF inoculation performed the best in 2017. All of these results suggested that our AMF and PSB could effectively promote growth and production of sunchoke under field conditions. Such effects were varied due to different environmental conditions each year. Note that this is the first study showing successful co-inoculation of AMF and PSB for promoting growth and yield of sunchoke in the real cultivation fields.

Biosurfactant based formulation of Pseudomonas guariconensis LE3 with multifarious plant growth promoting traits controls charcoal rot disease in Helianthus annus.

Biosurfactants are environment compatible surface-active biomolecules with multifunctional properties which can be utilized in various industries. In this study a biosurfactant producing novel plant growth promoting isolate Pseudomonas guariconensis LE3 from the rhizosphere of Lycopersicon esculentum is presented as biostimulant and biocontrol agent. Biosurfactant extracted from culture was characterized to be mixture of various mono- and di-rhamnolipids with antagonistic activity against Macrophomina phaseolina, causal agent of charcoal rot in diverse crops. Fourier transform infrared spectroscopy (FTIR) and proton nuclear magnetic resonance (1H NMR) analysis confirmed the rhamnolipid nature of biosurfactant. PCR analysis established the presence of genes involved in synthesis of antibiotics diacetylphloroglucinol, phenazine 1-carboxylic acid and pyocyanin, and lytic enzymes chitinase and endoglucanase suggesting biocontrol potential of the isolate. Plant growth promoting activities shown by LE3 were phosphate solubilization and production of siderophores, indole acetic acid (IAA), ammonia and 1-aminocyclopropane-1-carboxylate deaminase (ACCD). To assemble all the characteristics of LE3 various bioformuations were developed. Amendment of biosurfactant in bioformulation of LE3 cells improved the shelf life. Biosurfactant amended formulation of LE3 cells was most effective in biocontrol of charcoal rot disease of sunflower and growth promotion in field conditions. The root adhered soil mass of plantlets inoculated with LE3 plus biosurfactant was significantly higher over control. Biosurfactant amended formulation of LE3 cells caused maximum yield enhancement (80.80%) and biocontrol activity (75.45%), indicating that addition of biosurfactant improves the plant-bacterial interaction and soil properties leading to better control of disease and overall improvement of plant health and yield.

Development of a robust hydroponic method for screening of sunflower (Helianthus annuus L.) accessions for tolerance to heat and osmotic stress.

Hydroponic systems are known to provide a platform for uniform growth conditions until the reproductive stage. However, many plant species, including sunflower, show poor growth and survivability under conventional hydroponic systems due to poor nutrient availability, hypoxia and algal contamination. Thus, we tested various hydroponic systems to select a hydroponic system suitable for screening of sunflower germplasm. Sunflower accessions showed better growth and leaf gas exchange in newly-designed over conventional hydroponic systems. Selected hydroponic systems were further engaged in sunflower accession screening under heat and osmotic stress in a two-pan system (210 cm × 60 cm). Heat stress treatment was applied by growing sunflower germplasm at 42 °C and osmotic stress by adding polyethylene glycol 8000 which decreased the osmotic potential to - 0.6 MPa. There was significant variability among the sunflower accessions for their ability to survive under stress. Accessions such as C-2721 (43%), C-291 (46%) and D-14 (43%) had lower cell membrane injury percentage under osmotic stress and high seedling survivability (60‒80%) under heat stress when compared with susceptible accessions. Moreover, resistant accessions exhibited greater cuticular waxes and root length but lower transpiration losses. The newly designed hydroponic platform proved reliable for the selection of resistant sunflower accessions. Selected parental lines were validated by assessing their hybrids under field trials across two seasons under water and temperature stress during the reproductive phase (autumn). Hybrid H3 obtained by crossing drought and heat resistant parents had the highest seed yield and water use efficiency.

Gallic acid improves the antioxidant ability against cadmium toxicity: Impact on leaf lipid composition of sunflower (Helianthus annuus) seedlings.

In the present work, the effect of seed pre-soaking with gallic acid (GA; 3,4,5-triphydroxyl-benzoic acid) in conferring subsequent tolerance to Cd stress in sunflower (Helianthus annuus) seedlings was investigated. Exposing sunflower seedlings to increasing Cd concentrations (5, 10 and 20 µM) caused a gradual decrease in root and shoot biomass and increased the metal accumulation in both organs. Seed pretreatment with 75 µM GA significantly restricted Cd uptake, markedly alleviated Cd-induced plant growth inhibition, and mitigated the oxidative damages caused by this metal, as compared to plants directly exposed to Cd. GA pre-soaking prior to Cd stress also enhanced catalase, ascorbate peroxidase and glutathione reductase activities, while inhibiting that of superoxide dismutase. This was associated with increased levels of total thiols and glutathione along with a decreased level of oxidized glutathione in leaves. Moreover, GA pre-soaking led to changes in leaf fatty acid composition of seedlings challenged with Cd, as evidenced by the higher total lipid content and lipid unsaturation degree. As a whole, this study provides strong arguments highlighting the potential role of GA as a growth promoter for sunflower seedlings submitted to Cd stress, notably by boosting the antioxidant defense system and improving leaf membrane stability.

Pseudomonas entomophila PE3 and its exopolysaccharides as biostimulants for enhancing growth, yield and tolerance responses of sunflower under saline conditions.

Evaluation of plant growth promoting bacteria and the associated metabolites under saline conditions can be a potential eco-friendly remediation and productivity enhancement strategy. Salt-tolerant Pseudomonas entomophila PE3 was isolated from saline soil and screened for plant growth promoting (PGP) traits. The isolate produced indole acetic acid (IAA), gibberellic acid (GA), exopolysaccharides (EPS) and siderophore along with the potential to solubilize potassium (K), zinc (Zn) and phosphorus (P). Maximum stimulation of PGP attributes was recorded at 2% NaCl concentration. To determine the role of EPS, their composition was analyzed (at different salt concentrations) and comparison was done to determine the changes upon exposure to salinity. EPS was found to be rich in carbohydrates, proteins and phenolic compounds. The extracted EPS were also found to possess salt-tolerance properties including antioxidant, hydroxyl scavenging activity, reducing power, emulsification and flocculation potential, and Na+ accumulation ability. Interestingly, the salt tolerance properties of EPS were enhanced upon exposure to salinity (2% NaCl). Finally, EPS based bioformulation of isolate PE3 was checked through field assay in saline soil. With promising results on growth promotion and improved salinity tolerance attributes of inoculated sunflower plants, the bioformulation of PE3 amended with EPS can be a breakthrough for remediation of saline-agroecosystems.

Activated sludge treatment for promoting the reuse of a synthetic produced water in irrigation.

Large volumes of produced water are generated as a byproduct in activities of oil and gas exploitation, which can be reused in agriculture after a treatment process. Activated sludge treatment has been successfully used to remove oil from wastewater, but systematic studies on the toxicity of this effluent using this treatment are scarce in the literature. In this study, it was investigated the performance of an activated sludge system in the treatment of a synthetic produced water under different initial conditions in terms of salinity and oil and grease concentration. Furthermore, it was evaluated this effluent phytotoxicity in the germination, and seedling and plant growths of sunflower and corn seeds using untreated and treated synthetic produced water. Results revealed the activated sludge effectiveness in oil and grease and salinity removal from produced water, viz. high removal efficiency of 99.01 ± 0.28 and 91.07 ± 0.39%., respectively. Untreated produced water showed considerable toxic effects on the germination (74.67 ± 2.31% and 82.67 ± 2.31 for sunflower and corn seeds, respectively) and growth stages of sunflower and corn seed plants. The germination percentage was approximately 100% for both types of seed. The seedling and plant growth of the two seeds irrigated with treated produced water had similar performance when used tap water. These results highlighted the potential reuse as an unconventional water resource for plant irrigation of the synthetic produced water treated by an activated sludge process, which technology has showed high removal performance of salinity and oil.

Exogenous plant growth regulator alleviate the adverse effects of U and Cd stress in sunflower (Helianthus annuus L.) and improve the efficacy of U and Cd remediation.

Plant growth regulators (PGRs) are widely used in agricultural activities and have the potential to improve plant growth and plant tolerance against metal stress. PGR-assisted phytoextraction is now an effective and inexpensive method for enhancing the plant removal of toxic metals from soil. In this study, we conducted experiments to determine the effects of PGR treatments on soil uranium (U) and cadmium (Cd) removal by sunflowers as well as their stress response to U and Cd contamination. We found that the plant growth was inhibited by combined U and Cd stress in sunflowers compared with that of the control; however, the application of exogenous PGR had reduced the combined U and Cd stress by stimulating photosynthesis, decreasing the levels of active oxygen and lipid peroxidation, and enhancing the activity of the antioxidant defence systems. Exogenous PGR also increased the uptake of U and Cd by sunflowers and therefore, improved their U and Cd remediation efficiency. Moreover, indoleacetic acid (IAA) was the most effective PGR at removing U and Cd in the soil; the U and Cd removal efficiency was 484.21% and 238.85% higher in the 500 mg L-1 IAA application compared with that of the control without PGR application, respectively. Furthermore, none of the PGR treatments significantly influenced the available U and Cd contents in soil. Our results, therefore, may provide some detailed understanding on the technologies for the sustainable remediation of U and Cd contaminated soil by the combination of PGR treatments and phytoextraction.

Genetic Variation Analysis for Breeding Purposes in Wild and Cultivated Sunflower Germplasm (Helianthus annuus Sp.).

BACKGROUND AND OBJECTIVE: Molecular genetic methods show conclusive turn in phylogeny investigations and therefore, understanding the allocation and extension of genetic diversity inside and among species. The current study aims to evaluate genetic diversity using molecular markers (5 SSR and 5 ISSR) on 13 sunflowers (Helianthus annuus) genotypes from different regions. MATERIALS AND METHODS: The data acquired with the highest degree of polymorphism were confirmed. Polymorphic bands (24 and 55) were demonstrated totally through SSR and ISSR, respectively. Nevertheless, ISSR was superior in exhibiting polymorphism rate (97.37%) compared to SSR markers (90.83%). A range from 2-21 of amplified fragments, with molecular weights varying from 34-1331 bp, was noticeable. The genetic similarity coefficient was employed to set UPGMA dendrogram. RESULTS: Dissimilarity was found in genotypes clustering within groups, whilst the manner of clustering in genotypes kept adjacent in ISSR and aggregated results of SSR and ISSR. Dice similarity ranged from 0.058-0.792, 0.168-0.770 and 0.154-0.847 with SSR, ISSR and the combined phylogeny dendrogram, respectively. The genetic variance of sunflower accessions of wild and cultivated cultivars from different geographic regions was established. CONCLUSION: Knowledge of these results could be applied to confirm a wide genetic basis for outlook sunflower and manage germplasm breeding programs.

Ray flower initiation in the Helianthus radula inflorescence is influenced by a functional allele of the HrCYC2c gene.

The radiate pseudanthium, with actinomorphic disk flowers surrounded by showy marginal zygomorphic ray flowers, is the most common inflorescence in the Helianthus genus. In Helianthus radula, ray flower primordia are normally absent at the dorsal domain of the inner phyllaries (discoid heads) while the occurrence of radiate inflorescences is uncommon. In Helianthus spp., flower symmetry and inflorescence architecture are mainly controlled by CYCLOIDEA (CYC)-like genes but the putative role of these genes in the development of discoid inflorescences has not been investigate. Three CYC genes of H. radula with a role in ray flower identity (HrCYC2c, HrCYC2d, and HrCYC2e) were isolated. The phylogenetic analysis placed these genes within the CYC2 subclade. We identified two different alleles for the HrCYC2c gene. A mutant allele, designed HrCYC2c-m, shows a thymine to adenine transversion, which generates a TGA stop codon after a translation of 14 amino acids. We established homozygous dominant (HrCYC2c/HrCYC2c) and recessive (HrCYC2c-m/HrCYC2c-m) plants for this nonsense mutation. Inflorescences of both HrCYC2c/HrCYC2c and HrCYC2c/HrCYC2c-m plants initiated ray flowers, despite at low frequency. By contrast, plants homozygous for the mutant allele (HrCYC2c-m/HrCYC2c-m) failed at all to develop ray flowers. The results support, for the first time, a role of the HrCYC2c gene on the initiation of ray flower primordia. However, also in the two dominant phenotypes, discoid heads are the prevalent architecture suggesting that this gene is required but not sufficient to initiate ray flowers in pseudanthia. Other unknown major genes are most likely required in the shift from discoid to radiate inflorescence.

Massive haplotypes underlie ecotypic differentiation in sunflowers.

Species often include multiple ecotypes that are adapted to different environments1. However, it is unclear how ecotypes arise and how their distinctive combinations of adaptive alleles are maintained despite hybridization with non-adapted populations2-4. Here, by resequencing 1,506 wild sunflowers from 3 species (Helianthus annuus, Helianthus petiolaris and Helianthus argophyllus), we identify 37 large (1-100 Mbp in size), non-recombining haplotype blocks that are associated with numerous ecologically relevant traits, as well as soil and climate characteristics. Limited recombination in these haplotype blocks keeps adaptive alleles together, and these regions differentiate sunflower ecotypes. For example, haplotype blocks control a 77-day difference in flowering between ecotypes of the silverleaf sunflower H. argophyllus (probably through deletion of a homologue of FLOWERING LOCUS T (FT)), and are associated with seed size, flowering time and soil fertility in dune-adapted sunflowers. These haplotypes are highly divergent, frequently associated with structural variants and often appear to represent introgressions from other-possibly now-extinct-congeners. These results highlight a pervasive role of structural variation in ecotypic adaptation.

Salt stress induced differential metabolic responses in the sprouting tubers of Jerusalem artichoke (Helianthus tuberosus L.).

To better understand the mechanism of inherent salt resistance in Jerusalem artichoke (Helianthus tuberosus L.), physiological and metabolic responses of tubers at the initiation stage of sprouting under different salt stress levels were evaluated in the present study. As a result, 28 metabolites were identified using proton nuclear magnetic resonance (1H-NMR) spectroscopy. Jerusalem artichoke tubers showed minor changes in metabolic response under moderate salt stress when they had not yet sprouted, where metabolism was downregulated at the start of sprouting and then upregulated significantly after plants became autotrophic. However, mild and severe salt stress levels caused different metabolic response patterns. In addition, the accumulation of fructose and sucrose was enhanced by moderate salt stress, while glucose was highly consumed. Aspartate and asparagine showed accelerated accumulation in sprouting Jerusalem artichoke tubers that became autotrophic, suggesting the enhancement of photosynthesis by moderate salt stress.

Spatial and developmental synthesis of endogenous sesquiterpene lactones supports function in growth regulation of sunflower.

MAIN CONCLUSION: Tissue-specific occurrence and formation of endogenous sesquiterpene lactones has been assessed and suggests physiological function as antagonists of auxin-induced plant growth in sunflower. Sunflower, Helianthus annuus, accumulate high concentrations of bioactive sesquiterpene lactones (STL) in glandular trichomes, but in addition, structurally different STL occur in only trace amounts in the inner tissues. The spatial and temporal production of these endogenous STL during early phases of plant development is widely unknown and their physiological function as putative natural growth regulators is yet speculative. By means of HPLC and MS analysis it was shown that costunolide, dehydrocostuslactone, 8-epixanthatin and tomentosin are already present in dry seeds and can be extracted in low amounts from cotyledons, hypocotyls and roots of seedlings during the first days after germination. Semi-quantitative and RT-qPCR experiments with genes of the key enzymes of two independent routes of the endogenous STL biosynthesis confirmed the early and individual expression in these organs and revealed a gradual down regulation during the first 72-96 h after germination. Light irradiation of the plants led to a fast, but transient increase of STL in parts of the hypocotyl which correlated with growth retardation of the stem. One-sided external application of costunolide on hypocotyls conferred reduced growth of the treated side, thus resulting in the curving of the stem towards the side of the application. This indicates the inhibiting effects of STL on plant growth. The putative function of endogenous STL in sunflower as antagonists of auxin in growth processes is discussed.