MsSAMS, a cold stress-responsive gene, provides resistance to environmental stress in T2-generation transgenic plants.

The SAMS (S-adenosylmethionine synthetase) gene is known to play an important role in the mechanism of cold resistance, as overexpression of this gene results in phenotypic changes in T1-generation transgenic plants. Accordingly, this study was conducted to test the expression of the MsSAMS gene in T2-generation transgenic plants and to investigate the resistance of these plants and the function of the transgene in response to various environmental stresses. For the morphological analysis of T2-generation transgenic plants overexpressing the MsSAMS gene, observations using scanning electron microscopy (SEM) were performed. T2-generation transgenic plants were obtained by planting a total of 5 lines, and their characteristics were tested by comparisons with those of the control. SEM revealed that the thickest leaves were produced by the T6 transgenic line-161.24 ± 8.05 µm. The number of stomata ranged from 20.00 ± 2.65 to 34.00 ± 1.00 in the T2-generation transgenic plants, but the control had more stomata. Resistance to various factors, such as low temperature, drought, and oxidative stress, in the T2-generation transgenic plants was also confirmed. Under cold-stress conditions, the T6 transgenic line presented the lowest value (22.73%) of ion leakage, and under drought-stress conditions, compared with the control, the transgenic lines presented lower ion leakage after being treated with various concentrations of mannitol. Even under oxidative-stress conditions, the T2-generation transgenic plants presented ion leakage levels that were 32.91 ± 4.24 to 48.33 ± 3.54% lower than those of the control after treatment with various concentrations of methyl viologen. Regarding SAMS enzyme activity, as the duration of cold treatment increased, the activity in the transgenic plants tended to decrease and then increase. During 48 h of cold treatment, the control showed a decrease in SAM content, while the T2-generation transgenic plants presented an increase in SAM content, from 13.58 ± 1.04 to 22.75 ± 1.95 mg protein/g FW. The results suggest that the MsSAMS gene may be important to the mechanisms of resistance to oxidative and drought stresses in addition to its previously known association with cold resistance. Based on these results, it was suggested that the MsSAMS gene, whose expression is induced by cold stress, can serve as a marker of various responses to environmental stresses, because resistance to cold damage and various environmental stresses are stably inherited in the T2 generation.

Comparative Study on Seed Characteristics, Antioxidant Activity, and Total Phenolic and Flavonoid Contents in Accessions of Sorghum bicolor (L.) Moench.

Sorghum is a major cereal food worldwide, and is considered a potential source of minerals and bioactive compounds. Its wide adaptive range may cause variations in its agronomic traits, antioxidant properties, and phytochemical content. This extensive study investigated variations in seed characteristics, antioxidant properties, and total phenolic (TPC) and flavonoid contents (TFC) of sorghum collected from different ecological regions of 15 countries. The antioxidant potential of the seed extracts of various sorghum accessions was determined using 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azinobis 3-ethylbenzothiazoline-6-sulfonate (ABTS) radical scavenging assays. Significant variations in TPC were observed among the sorghum accessions. All 78 sorghum accessions used in this study exhibited significant variations in TFC, with the lowest and highest amount observed in accessions C465 and J542, respectively. DPPH scavenging potential of the seed extracts for all the accessions ranged from 11.91 ± 4.83 to 1343.90 ± 81.02 µg mL-1. The ABTS assay results were similar to those of DPPH but showed some differences in the accessions. Pearson's correlation analysis revealed a wide variation range in the correlation between antioxidant activity and TPC, as well as TFC, among the sorghum accessions. A wide diversity range was also recorded for the seed characteristics (1000-seed weight and seed germination rate). A dendrogram generated from UPGMA clustering, based on seed traits, antioxidant activity, TPC, and TFC was highly dispersed for these accessions. Variations among the accessions may provide useful information regarding the phytoconstituents, antioxidant properties, and phytochemical contents of sorghum and aid in designing breeding programs to obtain sorghum with improved agronomic traits and bioactive properties.

Application of Light-Emitting Diodes for Improving the Nutritional Quality and Bioactive Compound Levels of Some Crops and Medicinal Plants.

Light is a key factor that affects phytochemical synthesis and accumulation in plants. Due to limitations of the environment or cultivated land, there is an urgent need to develop indoor cultivation systems to obtain higher yields with increased phytochemical concentrations using convenient light sources. Light-emitting diodes (LEDs) have several advantages, including consumption of lesser power, longer half-life, higher efficacy, and wider variation in the spectral wavelength than traditional light sources; therefore, these devices are preferred for in vitro culture and indoor plant growth. Moreover, LED irradiation of seedlings enhances plant biomass, nutrient and secondary metabolite levels, and antioxidant properties. Specifically, red and blue LED irradiation exerts strong effects on photosynthesis, stomatal functioning, phototropism, photomorphogenesis, and photosynthetic pigment levels. Additionally, ex vitro plantlet development and acclimatization can be enhanced by regulating the spectral properties of LEDs. Applying an appropriate LED spectral wavelength significantly increases antioxidant enzyme activity in plants, thereby enhancing the cell defense system and providing protection from oxidative damage. Since different plant species respond differently to lighting in the cultivation environment, it is necessary to evaluate specific wavebands before large-scale LED application for controlled in vitro plant growth. This review focuses on the most recent advances and applications of LEDs for in vitro culture organogenesis. The mechanisms underlying the production of different phytochemicals, including phenolics, flavonoids, carotenoids, anthocyanins, and antioxidant enzymes, have also been discussed.

Neuroprotective Effect and Antioxidant Potency of Fermented Cultured Wild Ginseng Root Extracts of Panax ginseng C.A. Meyer in Mice.

Wild ginseng has better pharmacological effects than cultivated ginseng. However, its industrialization is limited by the inability to grow wild ginseng on a large scale. Herein, we demonstrate how to optimize ginseng production through cultivation, and how to enhance the concentrations of specific ginsenosides through fermentation. In the study, we also evaluated the ability of fermented cultured wild ginseng root extract (HLJG0701-ß) to inhibit acetylcholinesterase (AChE), as well as its neuroprotective effects and antioxidant activity. In invitro tests, HLJG0701-ß inhibited AChE activity and exerted neuroprotective and antioxidant effects (showing increased catalyst activity but decreased reactive oxygen species concentration). In invivo tests, after HLJG0701-ß was orally administered at doses of 0, 125, 250, and 500 mg/kg in an animal model of memory impairment, behavioral evaluation (Morris water maze test and Y-maze task test) was performed. The levels of AChE, acetylcholine (ACh), blood catalase (CAT), and malondialdehyde (MDA) in brain tissues were measured. The results showed that HLJG0701-ß produced the best results at a dose of 250 mg/kg or more. The neuroprotective mechanism of HLJG0701-ß was determined to involve the inhibition of AChE activity and a decrease in oxidative stress. In summary, both invitro and invivo tests confirmed that HJG0701-ß administration can lead to memory improvement.

Population structure of Miscanthus sacchariflorus reveals two major polyploidization events, tetraploid-mediated unidirectional introgression from diploid M. sinensis, and diversity centred around the Yellow Sea.

BACKGROUND AND AIMS: Miscanthus, a C4 perennial grass native to East Asia, is a promising biomass crop. Miscanthus sacchariflorus has a broad geographic range, is used to produce paper in China and is one of the parents (along with Miscanthus sinensis) of the important biomass species Miscanthus × giganteus. The largest study of M. sacchariflorus population genetics to date is reported here. METHODS: Collections included 764 individuals across East Asia. Samples were genotyped with 34 605 single nucleotide polymorphisms (SNPs) derived from restriction site-associated DNA sequencing (RAD-seq) and ten plastid microsatellites, and were subjected to ploidy analysis by flow cytometry. KEY RESULTS: Six major genetic groups within M. sacchariflorus were identified using SNP data: three diploid groups, comprising Yangtze (M. sacchariflorus ssp. lutarioriparius), N China and Korea/NE China/Russia; and three tetraploid groups, comprising N China/Korea/Russia, S Japan and N Japan. Miscanthus sacchariflorus ssp. lutarioriparius was derived from the N China group, with a substantial bottleneck. Japanese and mainland tetraploids originated from independent polyploidization events. Hybrids between diploid M. sacchariflorus and M. sinensis were identified in Korea, but without introgression into either parent species. In contrast, tetraploid M. sacchariflorus in southern Japan and Korea exhibited substantial hybridization and introgression with local diploid M. sinensis. CONCLUSIONS: Genetic data indicated that the land now under the Yellow Sea was a centre of diversity for M. sacchariflorus during the last glacial maximum, followed by a series of migrations as the climate became warmer and wetter. Overall, M. sacchariflorus has greater genetic diversity than M. sinensis, suggesting that breeding and selection within M. sacchariflorus will be important for the development of improved M. × giganteus. Ornamental M. sacchariflorus genotypes in Europe and North America represent a very narrow portion of the species' genetic diversity, and thus do not well represent the species as a whole.

Assessment of Diversity in the Accessions of Setaria italica L. Based on Phytochemical and Morphological Traits and ISSR Markers.

This study was carried out to evaluate genetic diversity, phenolic compound composition, and biological activity of Setaria italica L. collected from different parts of South Korea. Antioxidant potential of seeds was estimated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging assay, and antimicrobial activity was determined by evaluating the minimum inhibitory concentration (MIC). Eight phenolic acids and 3 flavonoids were identified and quantified, among which myricetin and salicylic acid were the most dominant phytochemical compounds detected in the majority of accessions. The antioxidant potential of the leaf extracts of all the accessions was significantly higher (ranging from 32.33 ± 1.53 µg mL-¹ in SI-03 to 87.87 ± 1.63 µg mL-¹) in SI-10 than that of the root, stem, or seeds. Among the 15 accessions, methanolic extracts of the SI-15 accession strongly suppressed the growth of Escherichia coli (250 µg mL-¹). Accessions SI-14 and SI-15 showed positive antimicrobial activity against all gram-positive bacteria. Interestingly, extracts of all accessions were more sensitive towards E. coli and Staphylococcus aureus, with MICs ranging from 250 to 1000 µg mL-¹. Three phenolic acids, namely chlorogenic acid, catechin, caffeic acid, naringin, hesperetin, and myricetin, were found to be moderately positively correlated with antioxidant activities. A wide range of diversity was observed in morphological traits, namely plant height (99.33 to 201.33 cm), culm length (67.10 to 160.00 cm), spike length (12.80 to 24.00 cm), 1000 seeds weight 1.44 to 2.91 g), bloom beginning (93.67 to 128.00 days), and full bloom (99.67 to 135 days). A dendogram generated from unweighted pair group method with arithmetic mean clustering (UPGMA) cluster analysis based on the morphological traits and inter simple sequence repeat (ISSR) marker data revealed three major groups. However, no clear correlation between these two different approaches was found. The average Shannon's information index value (I) was 0.492, and it ranged from 0 to 0.693. The average expected heterozygosity (He) was 0.335, and it ranged from 0 to 0.499. The substantial variation in the morphological traits, bioactive properties, and genetic diversity among the accessions may provide useful information for breeding programs attempting to obtain S. italica with improved bioactive properties.

Diversity in Accessions of Panicum miliaceum L. Based on Agro-Morphological, Antioxidative, and Genetic Traits.

The genetic diversity and antioxidant potential of Panicum miliaceum L. accessions collected from different geo-ecological regions of South Korea were evaluated and compared. Antioxidant potential of seeds was estimated by the 1,1-diphenyl-2-picrylhydrazyl (DPPH) and 2,2'-azino-bis-3-ethylbenzthiazoline-6-sulfonic acid (ABTS) radical scavenging assays and total phenolic content was determined by the Folin⁻Ciocalteu method. Total phenolic content (TPC) in 80% methanolic extracts ranged from 16.24 ± 0.86 to 58.04 ± 1.00 mg gallic acid equivalent (GAE)/g of the sample extracts and total flavonoid content (TFC) varied from 7.19 ± 1.05 to 52.56 ± 1.50 mg quercetin equivalents (QE) mg/g of the sample extracts. DPPH radical scavenging capacity of the extracts from the 15 accessions of P. miliaceum varied from 206.44 ± 7.72 to 2490.24 ± 4.641 mg GAE/g of the sample extracts and ABTS radical scavenging capacity ranged from 624.85 ± 13.1 to 1087. 77 ± 9.58 mg GAE/g of the sample extracts. A wide range of genetic variation was observed as measured by Shannon's information index (I), number of effective alleles (Ne), number of observed alleles (Na), expected heterozygosity (He), unbiased expected heterozygosity (uHe). The observed variation in the bioactive properties, morphological traits, and genetic diversity among the accessions may provide useful information for breeding programs seeking to improve bioactive properties of P. miliaceum.

Ecological characteristics and in situ genetic associations for yield-component traits of wild Miscanthus from eastern Russia.

Background and aims Miscanthus is a genus of perennial C4 grasses native to East Asia. It includes the emerging ligno-cellulosic biomass crop M. ×giganteus, a hybrid between M. sinensis and M. sacchariflorus. Biomass yield and cold tolerance are of particular interest in Miscanthus, given that this crop is more temperate adapted than its C4 relatives maize, sorghum and sugarcane. Methods A plant exploration was conducted in eastern Russia, at the northern extreme of the native range for Miscanthus, with collections including 174 clonal germplasm accessions (160 M. sacchariflorus and 14 M. sinensis) from 47 sites. Accessions were genotyped by restriction site-associated DNA sequencing (RAD-seq) and plastid microsatellites. Key Results Miscanthus sinensis was found in maritime climates near Vladivostok (43·6°N) and on southern Sakhalin Island (46·6°N). Miscanthus sacchariflorus was found inland at latitudes as high as 49·3°N, where M. sinensis was absent. Most M. sacchariflorus accessions were diploid, but approx. 2 % were tetraploids. Molecular markers revealed little population structure (Jost's D < 0·007 among diploid groups) but high genetic diversity (expected heterozygosity = 0·14) within the collection of Russian M. sacchariflorus. Genome-wide association (GWA) analysis for traits measured at the collection sites revealed three M. sacchariflorus single nucleotide polymorphisms (SNPs) significantly associated with the number of stems per unit area, one with height and one with basal stem diameter; three were near or within previously described sorghum quantitative trait loci for related traits. Conclusions This new Miscanthus germplasm collection from eastern Russia will be useful for breeding Miscanthus and sugarcane cultivars with improved adaptation to cold. Moreover, a strategy is proposed to facilitate the rapid utilization of new germplasm collections: by implementing low-cost SNP genotyping to conduct GWA studies of phenotypic data obtained at collection sites, plant breeders can be provided with actionable information on which accessions have desirable traits and alleles.

Contrasting allelic distribution of CO/Hd1 homologues in Miscanthus sinensis from the East Asian mainland and the Japanese archipelago.

The genus Miscanthus is a perennial C4 grass native to eastern Asia and is a promising candidate bioenergy crop for cool temperate areas. Flowering time is a crucial factor governing regional and seasonal adaptation; in addition, it is also a key target trait for extending the vegetative phase to improve biomass potential. Homologues of CONSTANS (CO)/Heading date 1(Hd1) were cloned from Miscanthus sinensis and named MsiHd1. Sequences of MsiHd1 homologues were compared among 24 wild M. sinensis accessions from Japan, 14 from China, and three from South Korea. Two to five MsiHd1 alleles in each accession were identified, suggesting that MsiHd1 consists of at least three loci in the Miscanthus genome. Verifying the open reading frame in MsiHd1, they were classified as putative functional alleles without mutations or non-functional alleles caused by indels. The Neighbor-Joining tree indicated that one of the multiple MsiHd1 loci is a pseudogene locus without any functional alleles. The pseudogene locus was named MsiHd1b, and the other loci were considered to be part of the MsiHd1a multi-locus family. Interestingly, in most Japanese accessions 50% or more of the MsiHd1a alleles were non-functional, whereas accessions from the East Asian mainland harboured only functional alleles. Five novel miniature inverted transposable elements (MITEs) (MsiMITE1-MsiMITE5) were observed in MsiHd1a/b. MsiMITE1, detected in exon 1 of MsiHd1a, was only observed in Japanese accessions and its revertant alleles derived from retransposition were predominantly in Chinese accessions. These differences in MsiHd1a show that the dependency on functional MsiHd1a alleles is different between accessions from the East Asian mainland and Japan.

Genetic structure of Miscanthus sinensis and Miscanthus sacchariflorus in Japan indicates a gradient of bidirectional but asymmetric introgression.

Unilateral introgression from diploids to tetraploids has been hypothesized to be an important evolutionary mechanism in plants. However, few examples have been definitively identified, perhaps because data of sufficient depth and breadth were difficult to obtain before the advent of affordable high-density genotyping. Throughout Japan, tetraploid Miscanthus sacchariflorus and diploid Miscanthus sinensis are common, and occasionally hybridize. In this study, 667 M. sinensis and 78 M. sacchariflorus genotypes from Japan were characterized using 20 704 SNPs and ten plastid microsatellites. Similarity of SNP genotypes between diploid and tetraploid M. sacchariflorus indicated that the tetraploids originated through autopolyploidy. Structure analysis indicated a gradient of introgression from diploid M. sinensis into tetraploid M. sacchariflorus throughout Japan; most tetraploids had some M. sinensis DNA. Among phenotypically M. sacchariflorus tetraploids, M. sinensis ancestry averaged 7% and ranged from 1-39%, with introgression greatest in southern Japan. Unexpectedly, rare (~1%) diploid M. sinensis individuals from northern Japan were found with 6-27% M. sacchariflorus ancestry. Population structure of M. sinensis in Japan included three groups, and was driven primarily by distance, and secondarily by geographic barriers such as mountains and straits. Miscanthus speciation is a complex and dynamic process. In contrast to limited introgression between diploid M. sacchariflorus and M. sinensis in northern China, selection for adaptation to a moderate maritime climate probably favoured cross-ploidy introgressants in southern Japan. These results will help guide the selection of Miscanthus accessions for the breeding of biomass cultivars.

A footprint of past climate change on the diversity and population structure of Miscanthus sinensis.

BACKGROUND AND AIMS: Miscanthus is a perennial C4 grass that is a leading potential feedstock crop for the emerging bioenergy industry in North America, Europe and China. However, only a single, sterile genotype of M. × giganteus (M×g), a nothospecies derived from diploid M. sinensis (Msi) and tetraploid M. sacchariflorus (Msa), is currently available to farmers for biomass production. To facilitate breeding of Miscanthus, this study characterized genetic diversity and population structure of Msi in its native range of East Asia. METHODS: A total of 767 accessions were studied, including 617 Msi from most of its native range in China, Japan and South Korea, and 77 ornamental cultivars and 43 naturalized individuals from the USA. Accessions were evaluated with 21 207 restriction site-associated DNA sequencing single nucleotide polymorphism (SNP) markers, 424 GoldenGate SNPs and ten plastid microsatellite markers. KEY RESULTS: Six genetic clusters of Msi from geographically distinct regions in Asia were identified. Genetic data indicated that (1) south-eastern China was the origin of Msi populations found in temperate eastern Asia, which is consistent with this area probably having been a refugium during the last glacial maximum (LGM); (2) Msi migrated directly from south-eastern China to Japan before migrating to the same latitudes in China and Korea, which is consistent with the known sequence of warming post-LGM; (3) ornamental Msi cultivars were derived from the southern Japan population, and US naturalized populations were derived from a sub-set of the ornamental cultivars; and (4) many ornamental cultivars previously described as Msi have hybrid ancestry from Msa and Msi, whereas US naturalized populations of Msi do not. CONCLUSIONS: Population structure of Msi was driven by patterns of warming since the LGM, and secondarily by geographical barriers. This study will facilitate germplasm conservation, association analyses and identification of potential heterotic groups for the improvement of Miscanthus as a bioenergy crop.

The effect of different LED wavelengths on the components and biosynthesis of isoflavonoid in sprout Astragalus membranaceus.

An artificial light source is the optimal element for studying the usability of the medicinal plant Astragalus membranaceus as a sprout vegetable. Based on artificial light source conditions, formononetin (FO) level was the highest (2.6 mg/L) in A. membranaceus exposed to white light emitting diode (LED) light, and calycosin (CA) level was the highest (3.09 mg/L) in the plant exposed to red LED light. According to the publicly available transcriptome data of LED-exposed sprout A. membranaceus LED, reference genes related to the content enhancement of FO, an isoflavone compound, and those related to the content enhancement of CA were selected. The expression patterns of these genes were assayed using qPCR. Among the genes related to FO enhancement, Gene-225190T showed the highest mRNA levels in cells of LED-white light-exposed sprout A. membranaceus; among the genes related to CA enhancement, Gene_042770T showed the highest expression under red LED light. Most genes related to the overall biosynthesis regulation of flavonoids of the upper concept of isoflavone were highly expressed in response to red LED light, and the transcriptional level of 4CL in response to red LED light was the highest. Based on these results, the artificial light sources that regulated the FO and CA contents in sprouts A. membranaceus were white and red LED lights, and the selected reference genes were capable of regulating isoflavone biosynthesis.

Antisense-overexpression of the MsCOMT gene induces changes in lignin and total phenol contents in transgenic tobacco plants.

Initially, we isolated the caffeic acid O-methyltransferase (COMT) gene from Miscanthus sinensis (accession number HM062766.1). Next, we produced transgenic tobacco plants with down-regulated COMT gene expression to study its control of total phenol and lignin content and to perform morphological analysis. These transgenic plants were found to have reduced PAL and ascorbate peroxidases expression, which are related to the phenylpropanoid pathway and antioxidant activity. The MsCOMT-down-regulated plants had decreased total lignin in the leaves and stem compared with control plants. Reduced flavonol concentrations were confirmed in MsCOMT-down-regulated transgenic plants. We also observed a morphological difference, with reduced plant cell number in transgenic plants harboring antisense MsCOMT. The transgenic tobacco plants with down-regulated COMT gene expression demonstrate that COMT plays a crucial role related to controlling lignin and phenol content in plants. Also, COMT activity may be related to flavonoid production in the plant lignin pathway.

Cellular Morphology and Transcriptome Comparative Analysis of Astragalus membranaceus Bunge Sprouts Cultured In Vitro under Different LED Light.

Astragalus membranaceus, the major components of which are saponins, flavonoids, and polysaccharides, has been established to have excellent pharmacological activity. After ginseng, it is the second most used medicinal plant. To examine the utility of A. membranaceus as a sprout crop for plant factory cultivation, we sought to establish a functional substance control model by comparing the transcriptomes of sprouts grown in sterile, in vitro culture using LED light sources. Having sown the seeds of A. membranaceus, these were exposed to white LED light (continuous spectrum), red LED light (632 nm, 1.58 µmol/m2/s), or blue LED light (465 nm, 1.44 µmol/m2/s) and grown for 6 weeks; after which, the samples were collected for transcriptome analysis. Scanning electron microscopy analysis of cell morphology in plants exposed to the three light sources revealed that leaf cell size was largest in those plants exposed to red light, where the thickest stem was observed in plants exposed to white light. The total number of genes in A. membranaceus spouts determined via de novo assembly was 45,667. Analysis of differentially expressed genes revealed that for the comparisons of blue LED vs. red LED, blue LED vs. white LED, and red LED vs. white LED, the numbers of upregulated genes were 132, 148, and 144, respectively. Binding, DNA integration, transport, phosphorylation, DNA biosynthetic process, membrane, and plant-type secondary cell wall biogenesis were the most enriched in the comparative analysis of blue LED vs. red LED, whereas Binding, RNA-templated DNA biosynthetic process, DNA metabolic process, and DNA integration were the most enriched in the comparative analysis of blue vs. white LED, and DNA integration and resolution of meiotic recombination intermediates were the most enrichment in the comparison between red LED vs. white LED. The GO term associated with flavonoid biosynthesis, implying the functionality of A. membranaceus, was the flavonoid biosynthetic process, which was enriched in the white LED vs. red LED comparison. The findings of this study thus indicate that different LED light sources can differentially influence the transcriptome expression pattern of A. membranaceus sprouts, which can provide a basis for establishing a flavonoid biosynthesis regulation model and thus, the cultivation of high-functional Astragalus sprouts.

Impact of genotype-calling methodologies on genome-wide association and genomic prediction in polyploids.

Discovery and analysis of genetic variants underlying agriculturally important traits are key to molecular breeding of crops. Reduced representation approaches have provided cost-efficient genotyping using next-generation sequencing. However, accurate genotype calling from next-generation sequencing data is challenging, particularly in polyploid species due to their genome complexity. Recently developed Bayesian statistical methods implemented in available software packages, polyRAD, EBG, and updog, incorporate error rates and population parameters to accurately estimate allelic dosage across any ploidy. We used empirical and simulated data to evaluate the three Bayesian algorithms and demonstrated their impact on the power of genome-wide association study (GWAS) analysis and the accuracy of genomic prediction. We further incorporated uncertainty in allelic dosage estimation by testing continuous genotype calls and comparing their performance to discrete genotypes in GWAS and genomic prediction. We tested the genotype-calling methods using data from two autotetraploid species, Miscanthus sacchariflorus and Vaccinium corymbosum, and performed GWAS and genomic prediction. In the empirical study, the tested Bayesian genotype-calling algorithms differed in their downstream effects on GWAS and genomic prediction, with some showing advantages over others. Through subsequent simulation studies, we observed that at low read depth, polyRAD was advantageous in its effect on GWAS power and limit of false positives. Additionally, we found that continuous genotypes increased the accuracy of genomic prediction, by reducing genotyping error, particularly at low sequencing depth. Our results indicate that by using the Bayesian algorithm implemented in polyRAD and continuous genotypes, we can accurately and cost-efficiently implement GWAS and genomic prediction in polyploid crops.

Antioxidant, antimicrobial activity and inhibition of α-glucosidase activity by Betula alnoides Buch. bark extract and their relationship with polyphenolic compounds concentration.

Betula alnoides has been widely used in local traditional medicinal treatment for a variety of diseases, wounds and to cure diabetes. The air-dried, powdered (200 g) bark was extracted with 80% methanol at room temperature. The combined 80% methanolic extract was partitioned with organic solvents to yield n-hexane, ethyl acetate (EtOAc), n-butanol (BuOH; water saturated), and aqueous fractions for the investigation of antioxidant and antimicrobiology and α-glucosidase activity effects (GAE) of B. alnoides. Antioxidant activity was studied by using antioxidant tests, including electron donation ability test, reducing power, and metal-chelating activity assay. Antioxidant, antimicrobial activity and α-glucosidase inhibitory effect of 80% methanolic extracts and fractions derived from the bark of B. alnoides were evaluated and determined. The results showed that 80% methanolic extracts exhibited high 1,1-diphenyl-2-picrylhydrazyl (DPPH) scavenging activity (80.68%). In addition, both the 80% methanolic extract and EtOAc fraction exhibited more potent reducing activity than did butylated hydroxyanisole (BHA) and trolox. The aqueous fraction had higher metal-chelating activity than other fractions. The EtOAc fraction had the highest phenolic and flavonoid content (217.73 ± 1.02 mg GAE/g extract, and 38.42 ± 1.87 mg QE/g extract, respectively). The 80% methanolic extract and EtOAc fraction showed higher levels of antimicrobial activity than did other fractions. The 80% methanolic extract had the most powerful α-glucosidase inhibitory effect (98.46%) at a concentration of 40 µg/mL. The results suggest that bark extracts of B. alnoides could be a potential source of natural antioxidants and for treating pathogenic diseases.

Biochemical and Physiological Changes during Early Adventitious Root Formation in Chrysanthemum indicum Linné Cuttings.

Chrysanthemum indicum is an important ornamental and medicinal plant that is often difficult to propagate commercially because of its poor germination and low seed viability. This plant is mostly propagated by cutting, but the rooting is slow and non-uniform. The present investigation evaluated the regeneration capacity of stem cutting by examining the influence of auxins, growth medium, temperature, and explant type on adventitious root formation in C. indicum. The auxin-treated cuttings were planted in different growth substrates under greenhouse conditions. Among the different auxins tested, indole-3-butyric acid (IBA) more effectively induced roots. The cutting position of stock plants influenced rooting capacity. Cutting the stock plants from the apical region enhanced root number and length in the explants. Among the different explant types, apical stem cuts with 2000 ppm IBA produced a significantly higher number of adventitious roots when grown in vermiculite and perlite (V + P) at a ratio of 1:1 at 25 °C. High-performance liquid chromatography (HPLC) analysis revealed that protocatechuic acid, gentisic acid, chlorogenic acid, biochanin A, salicylic acid, caffeic acid, glycitein, and luteolin were the most dominant phenolic compounds present in C. indicum. These results indicate that IBA treatment promoted the synthesis and accumulation of phenolic compounds in C. indicum stem cuttings at the time of root formation. The present results demonstrate that applying auxins is essential for early root initiation and higher rooting success and thus may be beneficial for vegetative C. indicum propagation.

Managing flowering time in Miscanthus and sugarcane to facilitate intra- and intergeneric crosses.

Miscanthus is a close relative of Saccharum and a potentially valuable genetic resource for improving sugarcane. Differences in flowering time within and between Miscanthus and Saccharum hinders intra- and interspecific hybridizations. A series of greenhouse experiments were conducted over three years to determine how to synchronize flowering time of Saccharum and Miscanthus genotypes. We found that day length was an important factor influencing when Miscanthus and Saccharum flowered. Sugarcane could be induced to flower in a central Illinois greenhouse using supplemental lighting to reduce the rate at which days shortened during the autumn and winter to 1 min d-1, which allowed us to synchronize the flowering of some sugarcane genotypes with Miscanthus genotypes primarily from low latitudes. In a complementary growth chamber experiment, we evaluated 33 Miscanthus genotypes, including 28 M. sinensis, 2 M. floridulus, and 3 M. ×giganteus collected from 20.9° S to 44.9° N for response to three day lengths (10 h, 12.5 h, and 15 h). High latitude-adapted M. sinensis flowered mainly under 15 h days, but unexpectedly, short days resulted in short, stocky plants that did not flower; in some cases, flag leaves developed under short days but heading did not occur. In contrast, for M. sinensis and M. floridulus from low latitudes, shorter day lengths typically resulted in earlier flowering, and for some low latitude genotypes, 15 h days resulted in no flowering. However, the highest ratio of reproductive shoots to total number of culms was typically observed for 12.5 h or 15 h days. Latitude of origin was significantly associated with culm length, and the shorter the days, the stronger the relationship. Nearly all entries achieved maximal culm length under the 15 h treatment, but the nearer to the equator an accession originated, the less of a difference in culm length between the short-day treatments and the 15 h day treatment. Under short days, short culms for high-latitude accessions was achieved by different physiological mechanisms for M. sinensis genetic groups from the mainland in comparison to those from Japan; for mainland accessions, the mechanism was reduced internode length, whereas for Japanese accessions the phyllochron under short days was greater than under long days. Thus, for M. sinensis, short days typically hastened floral induction, consistent with the expectations for a facultative short-day plant. However, for high latitude accessions of M. sinensis, days less than 12.5 h also signaled that plants should prepare for winter by producing many short culms with limited elongation and development; moreover, this response was also epistatic to flowering. Thus, to flower M. sinensis that originates from high latitudes synchronously with sugarcane, the former needs day lengths >12.5 h (perhaps as high as 15 h), whereas that the latter needs day lengths <12.5 h.

Hovenia dulcis--an Asian traditional herb.

Hovenia dulcis Thunb., known as Japanese raisin tree, is commonly found in East Asia. It has a long history as a food supplement and traditional medicine in Japan, China and Korea, but is little known and used in Western countries so far. This minireview summarizes traditional uses and current knowledge on the pharmacology and phytochemistry of H. duclcis and covers, in particular, literature from specialized Asian journals that are not readily accessible. Extracts from H. dulcis accelerate detoxification of ethanol, and possess hepatoprotective, antioxidative, antimicrobial and antidiabetic properties. Although the underlying molecular mechanisms are not fully understood, free radical scavenging and enhancement of ethanol catabolism have been reported.

Screening of Allelochemicals in Miscanthus sacchariflorus Extracts and Assessment of Their Effects on Germination and Seedling Growth of Common Weeds.

There is increasing interest in the application of bioherbicides because they are less destructive to the global ecosystem than synthetic herbicides. Research has focused on reducing the dependence upon synthetic herbicides by substituting them with environmentally and economically sustainable bioproducts. Allelopathic phytochemicals may be an efficient method for controlling weeds, benefitting both the environment and human health. This study addressed the allelopathic potential of Miscanthus sacchariflorus (MS) extracts on the germination, plant growth, biomass, and biochemical parameters (electrolyte leakage, photosynthetic pigments, and antioxidant enzyme activities) of weeds using laboratory and field experiments. Liquid chromatography-mass spectrometry/mass spectrometry (LC-MS/MS) showed the presence of 22 phenolic compounds, including Orientin, Luteolin, Veratric acid, Chlorogenic acid, Protocatechuic acid, p-Coumaric acid, and Ferulic acid. Leaf extracts of M. sacchariflorus either completely suppressed or partially reduced seed germination and affected the development of weed seedlings (root and shoot length), in a dose-dependent manner. Aqueous extracts of M. sacchariflorus reduced the fresh weight and dry weight, affected the photosynthetic pigment content (chlorophylls, carotenoids), influenced the electrolyte ion leakage, and stimulated the activity of antioxidant enzymes in a species-specific manner. Pearson's correlation analysis showed that the phenolic compound composition of M. sacchariflorus correlated with the variables tested, indicating that the phytochemicals present in the plant extracts of M. sacchariflorus are a potential source of bio-herbicides.