A GWAS study highlights significant associations between a series of indels in a FLOWERING LOCUS T gene promoter and flowering time in white lupin (Lupinus albus L.).

BACKGROUND: White lupin (Lupinus albus L.) is a high-protein Old World grain legume with remarkable food and feed production interest. It is sown in autumn or early spring, depending on the local agroclimatic conditions. This study aimed to identify allelic variants associated with vernalization responsiveness, in order to improve our knowledge of legume flowering regulatory pathways and develop molecular selection tools for the desired phenology as required for current breeding and adaptation to the changing climate. RESULTS: Some 120 white lupin accessions originating from a wide range of environments of Europe, Africa, and Asia were phenotyped under field conditions in three environments with different intensities of vernalization, namely, a Mediterranean and a subcontinental climate sites of Italy under autumn sowing, and a suboceanic climate site of France under spring sowing. Two hundred sixty-two individual genotypes extracted from them were phenotyped in a greenhouse under long-day photoperiod without vernalization. Phenology data, and marker data generated by Diversity Arrays Technology sequencing (DArT-seq) and by PCR-based screening targeting published quantitative trait loci (QTLs) from linkage map and newly identified insertion/deletion polymorphisms in the promoter region of the FLOWERING LOCUS T homolog, LalbFTc1 gene (Lalb_Chr14g0364281), were subjected to a genome-wide association study (GWAS). Population structure followed differences in phenology and isolation by distance pattern. The GWAS highlighted numerous loci significantly associated with flowering time, including four LalbFTc1 gene promoter deletions: 2388 bp and 2126 bp deletions at the 5' end, a 264 bp deletion in the middle and a 28 bp deletion at the 3' end of the promoter. Besides LalbFTc1 deletions, this set contained DArT-seq markers that matched previously published major QTLs in chromosomes Lalb_Chr02, Lalb_Chr13 and Lalb_Chr16, and newly discovered QTLs in other chromosomes. CONCLUSIONS: This study highlighted novel QTLs for flowering time and validated those already published, thereby providing novel evidence on the convergence of FTc1 gene functional evolution into the vernalization pathway in Old World lupin species. Moreover, this research provided the set of loci specific for extreme phenotypes (the earliest or the latest) awaiting further implementation in marker-assisted selection for spring- or winter sowing.

Plant In Vitro Cultures of Coleus scutellarioides (L.) Benth. "Electric Lime" and Possibilities of Modification in the Biosynthesis of Volatile Compounds.

Coleus scutellarioides (L.) Benth. is a globally spread species, known for its characteristic spectacularly colorful leaves of decorative value. Thanks to its rich chemical composition, the plant is used in ethnopharmacology, and it is also regarded as having high medicinal potential. The application of in vitro cultures enables the acquisition of homogeneous certified material of high quality. Additionally, excluding the effect of biotic and abiotic factors on the plants is a way to fully recognize the influence of phytohormones on the plant morphology and the biosynthetic pathways of compound production. The best way to grow C. scutellarioides "Electric Lime" under in vitro conditions is to use the basic MS medium (Murashige and Skoog medium), enriched with naphthyl-1-acetic acid at a concentration of 0.5 mg dm-3. The analysis of volatile compounds demonstrated that the content of volatile compounds in the plants cultivated under in vivo conditions was expressed at a level of 2848.59 µg g-1, whereas in the plants bred in vitro without supplementation with phytohormones, the level was 8191.47 µg g-1. The highest content was noted for copaene, α-pinene, 1-octene-3-ol, α-selinene, sabinen, γ- and δ-cadinene, 3-octanol, and ß-pinene. Aroma profiling revealed a lack of boranyl acetate, 2-hexenal, and 2-hexen-1-ol in the plants cultivated under in vivo conditions. Differences were found in the volatile composition between plants bred in vivo and in vitro, with the most significant recorded for the contents of 1-octen-3-ol and 3-octanol. The addition of plant growth regulators into the basic medium under in vitro conditions affected the percentage ratio and contents of specific compounds in plant tissues. The most intense biosynthesis of volatile compounds took place in the plants cultivated on the medium enriched with NAA at 10,579.11 µg g-1, whereas the least intense was noted for plants cultivated on the medium supplemented with BA, where it was recorded at the level of 5610.02 µg g-1. So far, there has been no research published which would pertain to the profiling of volatile compounds performed using the SPME (solid-phase microextraction) technique. Moreover, the very few studies conducted on the chemical composition of these compounds do not mention the specific variety of C. scutellarioides under analysis.

The influence of cultivation conditions on the formation of psychoactive salvinorin A, salvinorin B, rosmarinic acid and caffeic acid in Coleus scutellarioides.

Coleus scutellarioides (L.) Benh. is a popular species in the world, known for its characteristic magnificent colourful leaves. The study has revealed that the contents of rosmarinic acid and caffeic acid are significantly higher in the plant tissues cultivated in vivo than when under in vitro conditions. The performed qualitative and quantitative analyses confirmed the presence (whose averaged content) of salvinorin A (6.65 µg/1 g of fresh plant) and salvinorin B (50.46 µg/1 g of fresh plant) in tissues of Coleus scutellarioides (L.) Benh. of 'Electric lime' variety. The greatest quantities of these compounds were recorded for plants cultivated in vitro on the MS medium enriched with NAA (naphthyl-1-acetic acid) at a concentration of 0.5 mg∙ dm-3. The research detected differences in the amounts of compounds between plants grown in vivo and those cultivated in vitro. Addition of plant growth regulators into the breeding medium under in vitro conditions was found affecting the amounts of compounds in plant tissues.

Cladophora glomerata extracts produced by Ultrasound-Assisted Extraction support early growth and development of lupin (Lupinus angustifolius L.).

The effect of the extract obtained by Ultrasound-Assisted Extraction (UAE) from green macroalga Cladophora glomerata on the germination and early growth of three narrow-leaved lupin varieties (cv. Homer, Jowisz, and Tytan) was examined. The seeds of these varieties came from five growing seasons (2015-2019) and this was their successive propagation stage. In total, 45 groups were tested. Narrow-leaved lupin like other legumes have a beneficial effect on the physical properties and fertility of the soil. Its high nutritive value makes it suitable for the production of valuable fodder. The algal extract, which was screened for the content of active compounds responsible for their biostimulant effect was applied in two concentrations: 10 and 20%. The germination percentage, root, hypocotyl, epicotyl length and chlorophyll content in cotyledons were evaluated at the end of the experiment. The 20% extract stimulated the growth of seedlings of all lupin cultivars better than the 10% application. The Jowisz variety deserves special attention, as it has the longest root system of seedlings.

Chemistry behind Quality-Emission of Volatile Enantiomers from Mentha spp. Plant Tissue in Relationship to Odor Sensory Quality.

The quality of food, considering increasing consumer demands and competition among producers, is a highly important issue. Quality concerns are also applicable to the odor quality of herbs and spices (HSs). Meanwhile, HSs commonly are graded based on their essential oils (EOs) content and analysis; but does the instrumental analysis really provide general information about the HSs sensory quality? Three chemotypes of Mentha spp. were used in the present study. From samples diversified by convective drying at different temperatures, EOs were hydrodistillated and analyzed by enantioselective GC-MS; moreover, the source plant material's volatile profile was analyzed by the HS-SPME technique. The instrumental analysis was confronted with the results of the sensory panel. Changes in enantiomeric composition were observed during the drying process, although no clear correlations or trends could be found for individual chiral components. Furthermore, even with significant differences in particular volatiles' contribution to plants' EOs and their volatile profiles, judges were not able to match the sample EOs and plant samples with sufficient effectiveness (~40%). Based on those results, we suggest that volatile enantiomeric distribution does not have an actual influence on odor quality and that the sensory analysis should not be replaced with instrumental analysis, which cannot predict general sensory quality.

The Potential Use of Plant Growth Regulators for Modification of the Industrially Valuable Volatile Compounds Synthesis in Hylocreus undatus Stems.

The pitaya (dragon fruit) Hylocereus is a genus which belongs to the Cactaceae family. It is native to Mexico, occurring also in other regions of Central and South America. Pitaya fruit is mainly intended for consumption and for this reason the species is grown commercially. The fruit is a rich source of vitamins, biologically active compounds, and dietary fibre. Using in vitro culture can accelerate the process of reproduction and growth of pitaya plants. Profiling of volatile compounds contained in the stem of Hylocereus undatus was carried out using the SPME-GC-MS technique. The main compounds present were hexanal, 2-hexenal and 1-hexanol. The results showed differences in the occurrence of volatile compounds between plants grown in media with an addition of BA (6-benzylaminopurine) and IAA (indole-3-acetic acid), which have been used as plant growth regulators. Statistically significant differences between the contents of volatile compounds were observed in the case of 2-hexenal and 1-hexanol. The effect of BA on reducing the amount of volatile compounds was observed. However, introduction of IAA to the in vitro medium resulted in more compounds being synthesized. This study is the first to describe the volatile compounds in the pitaya stem. The results indicate that plant hormones are able to modify the profile of volatile compounds.

Genotoxic Evaluation of Fe3O4 Nanoparticles in Different Three Barley (Hordeum vulgare L.) Genotypes to Explore the Stress-Resistant Molecules.

Sustainable agricultural practices are still essential due to soil degradation and crop losses. Recently, the relationship between plants and nanoparticles (NPs) attracted scientists' attention, especially for applications in agricultural production as nanonutrition. Therefore, the present research was carried out to investigate the effect of Fe3O4 NPs at low concentrations (0, 1, 10, and 20 mg/L) on three genotypes of barley (Hordeum vulgare L.) seedlings grown in hydroponic conditions. Significant increases in seedling growth, enhanced chlorophyll quality and quantity, and two miRNA expression levels were observed. Additionally, increased genotoxicity was observed in seedlings grown with NPs. Generally, Fe3O4 NPs at low concentrations could be successfully used as nanonutrition for increasing barley photosynthetic efficiency with consequently enhanced yield. These results are important for a better understanding of the potential impact of Fe3O4 NPs at low concentrations in agricultural crops and the potential of these NPs as nanonutrition for barley growth and yield enhancement. Future studies are needed to investigate the effect of these NPs on the expression of resistance-related genes and chlorophyll synthesis-related gene expression in treated barley seedlings.

Quantitative Control of Early Flowering in White Lupin (Lupinus albus L.).

White lupin (Lupinus albus L.) is a pulse annual plant cultivated from the tropics to temperate regions for its high-protein grain as well as a cover crop or green manure. Wild populations are typically late flowering and have high vernalization requirements. Nevertheless, some early flowering and thermoneutral accessions were found in the Mediterranean basin. Recently, quantitative trait loci (QTLs) explaining flowering time variance were identified in bi-parental population mapping, however, phenotypic and genotypic diversity in the world collection has not been addressed yet. In this study, a diverse set of white lupin accessions (n = 160) was phenotyped for time to flowering in a controlled environment and genotyped with PCR-based markers (n = 50) tagging major QTLs and selected homologs of photoperiod and vernalization pathway genes. This survey highlighted quantitative control of flowering time in white lupin, providing statistically significant associations for all major QTLs and numerous regulatory genes, including white lupin homologs of CONSTANS, FLOWERING LOCUS T, FY, MOTHER OF FT AND TFL1, PHYTOCHROME INTERACTING FACTOR 4, SKI-INTERACTING PROTEIN 1, and VERNALIZATION INDEPENDENCE 3. This revealed the complexity of flowering control in white lupin, dispersed among numerous loci localized on several chromosomes, provided economic justification for future genome-wide association studies or genomic selection rather than relying on simple marker-assisted selection.

Sarracenia alata (Alph.Wood) Alph.Wood Microcuttings as a Source of Volatiles Potentially Responsible for Insects' Respond.

Rare carnivorous plants representing the genus Sarracenia are perceived as very interesting to scientists involved in various fields of botany, ethnobotany, entomology, phytochemistry and others. Such high interest is caused mainly by the unique capacity of Sarracenia spp. to attract insects. Therefore, an attempt to develop a protocol for micropropagation of the Sarracenia alata (Alph.Wood) Alph.Wood, commonly named yellow trumpets, and to identify the specific chemical composition of volatile compounds of this plant in vitro and ex vivo was undertaken. Thus, the chemical volatile compounds excreted by the studied plant to attract insects were recognized with the application of the headspace solid-phase microextraction (HS-SPME) coupled with the GC-MS technique. As the major volatile compounds (Z)-3-hexen-1-ol (16.48% ± 0.31), (E)-3-hexen-1-ol acetate (19.99% ± 0.01) and ß-caryophyllene (11.30% ± 0.27) were identified. Further, both the chemical assumed to be responsible for attracting insects, i.e., pyridine (3.10% ± 0.07), and whole plants were used in in vivo bioassays with two insect species, namely Drosophila hydei and Acyrthosiphon pisum. The obtained results bring a new perspective on the possibilities of cultivating rare carnivorous plants in vitro since they are regarded as a valuable source of bioactive volatile compounds, as including ones with repellent or attractant activity.

Photoperiod and Vernalization Control of Flowering-Related Genes: A Case Study of the Narrow-Leafed Lupin (Lupinus angustifolius L.).

Narrow-leafed lupin (Lupinus angustifolius L.) is a moderate-yielding legume crop known for its high grain protein content and contribution to soil improvement. It is cultivated under photoperiods ranging from 9 to 17 h, as a spring-sown (in colder locations) or as an autumn-sown crop (in warmer regions). Wild populations require a prolonged cold period, called vernalization, to induce flowering. The key achievement of L. angustifolius domestication was the discovery of two natural mutations (named Ku and Jul) conferring vernalization independence. These mutations are overlapping deletion variants in the promoter of LanFTc1, a homolog of the Arabidopsis thaliana FLOWERING LOCUS T (FT) gene. The third deletion, named here as Pal, was recently found in primitive germplasm. In this study, we genotyped L. angustifolius germplasm that differs in domestication status and geographical origin for LanFTc1 alleles, which we then phenotyped to establish flowering time and vernalization responsiveness. The Ku and Jul lines were vernalization-independent and early flowering, wild (ku) lines were vernalization-dependent and late flowering, whereas the Pal line conferred intermediate phenotype. Three lines representing ku, Pal, and Ku alleles were subjected to gene expression surveys under 8- and 16-h photoperiods. FT homologs (LanFTa1, LanFTa2, LanFTc1, and LanFTc2) and some genes selected by recent expression quantitative trait loci mapping were analyzed. Expression profiles of LanFTc1 and LanAGL8 (AGAMOUS-like 8) matched observed differences in flowering time between genotypes, highlighted by high induction after vernalization in the ku line. Moreover, these genes revealed altered circadian clock control in Pal line under short days. LanFD (FD) and LanCRLK1 (CALCIUM/CALMODULIN-REGULATED RECEPTOR-LIKE KINASE 1) were negatively responsive to vernalization in Ku and Pal lines but positively responsive or variable in ku, whereas LanUGT85A2 (UDP-GLUCOSYL TRANSFERASE 85A2) was significantly suppressed by vernalization in all lines. Such a pattern suggests the opposite regulation of these gene pairs in the vernalization pathway. LanCRLK1 and LanUGT85A2 are homologs of A. thaliana genes involved in the FLOWERING LOCUS C (FLC) vernalization pathway. Lupins, like many other legumes, do not have any FLC homologs. Therefore, candidate genes surveyed in this study, namely LanFTc1, LanAGL8, LanCRLK1, and LanUGT85A2, may constitute anchors for further elucidation of molecular components contributing to vernalization response in legumes.

Mentha piperita L. Micropropagation and the Potential Influence of Plant Growth Regulators on Volatile Organic Compound Composition.

Due to the industrial use of Mentha piperita L. (peppermint), it is important to develop an optimal method to obtain standardized plant material with specific quality parameters. In vitro cultures may allow the production of desirable odor-active compounds (OACs) and improve their share in the plant aroma profile. There are two types of explants that are commonly used, apical meristems and nodal segments. In this study, the best overall effects were shown to be produced by the combination of MS medium with the addition of 0.5 mg·dm-3 indolyl-3-butyric acid. In this case, a very high degree of rooting was found (97% for apical meristems, 100% for nodal meristems), lateral shoots were induced in 83% of both types of explant, and the content of OACs in the plant aroma profile increased significantly, especially menthofurolactone and cis-carvone oxide, responsible in this case for a characteristic mint-like aroma. The comparison of the volatile organic compounds (VOCs) obtained from plants of different origin by GC-MS showed no significant differences in their qualitative composition. Moreover, in-vitro-cultivated peppermint on a medium containing 0.5 mg·dm-3 2-isopentinloadenine and 0.1 mg·dm-3 indolyl-3-acetic acid showed significant amounts of menthofurolactone in its VOC composition.

Plant-Mediated Enantioselective Transformation of Indan-1-one and Indan-1-ol. Part 2.

The main purpose of this publication was to obtain the S-enantiomer of indan-1-ol with high enantiomeric excess and satisfactory yield. In our research, we used carrot callus cultures (Daucus carota L.), whereby the enzymatic system reduced indan-1-one and oxidized indan-1-ol. During the reaction of reduction, after five days, we received over 50% conversion, with the enantiomeric excess of the formed S-alcohol above 99%. In turn, during the oxidation of racemic indan-1-ol after 15 days, 36.7% of alcohol with an enantiomeric excess 57.4% S(+) remained in the reaction mixture. In addition, our research confirmed that the reactions of reduction and oxidation are competing reactions during the transformation of indan-1-ol and indan-1-one in carrot callus cultures.

Diversity of selected Lupinus angustifolius L. genotypes at the phenotypic and DNA level with respect to microscopic seed coat structure and thickness.

The paper investigates seed coat characteristics (as a percentage of overall seed diameter) in Lupinus angustifolius L., a potential forage crop. In the study ten L. angustifolius genotypes, including three Polish cultivars, two Australian cultivars, three mutants originated from cv. 'Emir', and one Belarusian and one Australian breeding line were evaluated. The highest seed coat percentage was recorded in cultivars 'Sonet' and 'Emir'. The lowest seed coat thickness percentage (below 20%) was noted for breeding lines 11257-19, LAG24 and cultivar 'Zeus' (17.87%, 18.91% 19.60%, respectively). Despite having low seed weight, the Australian line no. 11257-19 was characterized by a desirable proportion of seed coat to the weight of seeds. In general, estimation of the correlation coefficient indicated a tendency that larger seeds had thinner coats. Scanning Electron Microscopy images showed low variation of seed coat sculpture and the top of seeds covered with a cuticle. Most of the studied genotypes were characterized by a cristatepapillate seed coat surface, formed by elongated polygonal cells. Only breeding line no. 11267-19 had a different shape of the cells building the surface layer of the coat. In order to illustrate genetic diversity among the genotypes tested, 24 ISSR primers were used. They generated a total of 161 polymorphic amplification products in 10 evaluated narrow-leaved lupin genotypes.

Application of DNA markers linked to the potato H1 gene conferring resistance to pathotype Ro1 of Globodera rostochiensis.

Ninety-one potato genotypes (cultivars and breeding lines) selected as resistant or susceptible to pathotype Ro1 of Globodera rostochiensis were screened for the presence of two PCR markers, 0.14 and 0.76 kb in length. Both PCR markers were linked with the H1 gene, located at the distal end of the long arm of chromosome V, and were present in 88 to 100% of the resistant cultivars and breeding lines. The 0.76 kb PCR marker was detected in all resistant genotypes and in approximately 86% of susceptible breeding lines as well as in all susceptible cultivars. The 0.14 kb marker was detected in 88% of resistant breeding lines and in 94% of resistant cultivars. Most of the susceptible genotypes tested (91% of cultivars, but only 50% of breeding lines) did not show the presence of the 0.14 kb marker. We conclude that the 0.14 kb H1 marker is likely to be useful for the proper selection of potato genotypes resistant to the Ro1 pathotype of G. rostochiensis.