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Chinese cabbage is an important dietary source of numerous phytochemicals, including glucosinolates and anthocyanins. The selection and development of elite Chinese cabbage cultivars with favorable traits is hindered by a long breeding cycle, a complex genome structure, and the lack of an efficient plant transformation protocol. Thus, a protoplast transfection-based transformation method may be useful for cell-based breeding and functional studies involving Chinese cabbage plants. In this study, we established an effective method for isolating Chinese cabbage protoplasts, which were then transfected with the pCAMBIA1303 binary vector according to an optimized PEG-based method. More specifically, protoplasts were isolated following a 4 h incubation in a solution comprising 1.5% (v/v) cellulase, 0.25% (v/v) macerozyme, 0.25% (v/v) pectinase, 0.5 M mannitol, 15 mM CaCl2, 25 mM KCl, 0.1% BSA, and 20 mM MES buffer, pH 5.7. This method generated 7.1 × 106 protoplasts, 78% of which were viable. The gfp reporter gene in pCAMBIA1303 was used to determine the transfection efficiency. The Chinese cabbage protoplast transfection rate was highest (68%) when protoplasts were transfected with the 40 μg binary vector for 30 min in a solution containing 40% PEG. The presence of gusA and hptII in the protoplasts was confirmed by PCR. The methods developed in this study would be useful for DNA-free genome editing as well as functional and molecular investigations of Chinese cabbage.

Drought is the primary abiotic stress that limits yield of soybean (Glycine max (L.) Merr.). The study aimed to identify yield-related quantitative trait loci (QTLs) in soybeans using a population of 160 F4-derived lines from ‘Hutcheson’ × PI 471938 crosses, which were cultivated under rain-fed and irrigated conditions. Seed yield was determined based on a total of nine irrigated and five rain-fed environments over two years. Twenty and twenty-seven SSR markers associated with yield (p ≤ 0.05) were identified in the irrigated and rain-fed environments, respectively. Four markers accounted for 22% of the yield variation in the irrigated environments (IR-YLD) and five markers explained 34% of the yield variation in the rain-fed environments (RF-YLD). Two independent IR-YLD and RF-YLD QTLs on chromosome (Chr) 13 (LG-F) were mapped to the Satt395-Sat_074 interval (4.2 cM) and near Sat_375 (3.0 cM), which explained 8% (LOD = 2.6) and 17% (LOD = 5.5) of the yield variation, respectively. The lines homozygous for the Hutcheson allele at the IR-YLD QTL linked to Sat_074 averaged 100 kg ha−1 higher yield than the lines homozygous for the PI 471938 allele. At two independent RF-YLD QTLs on Chr 13 and Chr 17, the lines homozygous for the PI 471938 alleles were 74 to 101 kg ha−1 higher in yield than the lines homozygous for the Hutcheson alleles. Three of the five significant SSR markers associated with RF-YLD were located in a genomic region known for canopy-wilting QTLs, in which the favorable alleles were inherited from PI 471938. The identification of yield-QTLs under the respective rain-fed and irrigated environments provides knowledge regarding differential responses of yield under different irrigation conditions, which will be helpful in developing high-yielding soybean cultivars.

Calnexin (CNX) is an integral membrane protein that functions as a chaperone in the endoplasmic reticulum for the correct folding of proteins under stress conditions, rendering organisms tolerant under adverse conditions. Studies have investigated the cytogenetic effects of gamma irradiation (Ɣ-IR) on plants, but information on the molecular response under Ɣ-IR remains limited. Previously, we constructed a cDNA library of an irradiation-sensitive bioindicator plant, Tradescantia BNL4430 (T-4430) under Ɣ-IR, in which the Calnexin-1 gene was highly upregulated at 50 mGy treatment. TrCNX1 encodes a 61.4 kDa protein with conserved signature motifs similar to already reported CNX1s. TrCNX1 expression was evaluated by semiquantitative reverse transcriptase PCR and quantitative real-time PCR and was ubiquitously expressed in various tissues and highly upregulated in flower petals under 50 mGy Ɣ-IR stress. The protective function of TrCNX1 was investigated by overexpression of TrCNX1 in an Escherichia coli BL21(DE3) heterologous system. Using plate assay, we showed that TrCNX1 increased the viability of E. coli transformants under both UV-B and Ɣ-IR compared with the control, demonstrating that TrCNX1 functions under irradiation stress. TrCNX1 may enhance irradiation stress tolerance in crops and act as a radio marker gene to monitor the effects of radiation.

Amino acids in the plant were intermediate metabolites which produced by uptake and assimilation of nitrogen and these extracts which gained by bio-chemical digestion from protein of plant or animal were a source of functional fertilizer. This study was conducted to evaluate effects of keratin amino acid fertilizer (KAF) gained from animal hair or hoof on changes of turfgrass quality and growth by investigating turf color index, chlorophyll index, shoot number, clipping yield, and nutrient content in the turfgrass tissue. Treatments were designed as follows; non-fertilizer (NF), compound fertilizer (CF), keratin amino acid fertilizer treatments [CF + KAF 0.26 ml m −2 (CKF), CF + KAF 0.52 ml m −2 (2CKF)], and only keratin amino acid fertilizer treatment (KF). Shoot number, clipping yield, and nitrogen uptake of KF were higher than those of CF. As compared with CF, soil chemical properties, turf color index, chlorophyll index and clipping yield of keratin amino acid fertilizer were not significant, but shoot number and uptake of N and K were increased significantly. These results show that the application of keratin amino acid fertilizer increased shoot number and growth by increased uptake of nitrogen and potassium.

Superintendents have used a silicate fertilizer to improve a resistance of turfgrass against several diseases, drought damage and wear stress. This study was conducted to evaluate the effect of liquid fertilizer containing silicate (LFSi) on changes of turfgrass quality and growth by investigating visual quality, chlorophyll content-chlorophyll a, chlorophyll b, and total chlorophyll, root length, shoot length, dry weight of clipping, and nutrient content in leaves tissue. Treatments were designed as follows; control fertilizer (CF), SiF-1 (CF + 1 ml m −2 LFSi), SiF-2 (CF + 2 ml m −2 LFSi), and SiF-3 (CF + 4 ml m −2 LFSi). As compared with CF, soil chemical properties, visual turfgrass quality, chlorophyll content, and dry weight of clipping of LFSi treatments were not significantly. Contrastingly, shoot density, root length, and the content of nitrogen or potassium were increased by application of LFSi. The content of Si in the tissue was positively correlated with potassium content or shoot length, and similarly shoot density positively with chlorophyll content or visual quality, respectively. These results suggested that the application of LFSi improved the turfgrass quality by increasing shoot density or K content in leaf tissue of creeping bentgrass.

Tetragonia tetragonoides (Pall.) Kuntze, called New Zealand spinach (NZS), is an edible plant used in salad in Western countries and has been used to treat gastrointestinal diseases in traditional medicine. We examined the anti-obesity and anti-hyperuricemic effects of NZS and the underlying mechanisms in high-fat diet (HFD)-induced obese mice. Mice were fed a normal-fat diet (NFD), high-fat diet (HFD), HFD with 75, 150, or 300 mg/kg NZS extract, or 245 mg/kg Garcinia cambogia (GC) extract. NZS decreased body weight gain, total white adipose tissue (WAT), liver weight, and size of adipocytes and improved hepatic and plasma lipid profiles. With NZS, the plasma levels of the leptin and uric acid were significantly decreased while the levels of the adiponectin were increased. Furthermore, NZS decreased the expression levels of adipogenesis-related genes and xanthine oxidoreductase (XOR), which is involved in uric acid production, while increasing that of proteins associated with fatty acid oxidation. UPLC analysis revealed that NZS contained 6-methoxykaempferol-3-O-&beta, d-glucosyl(1&prime, &prime, &rarr, 2&prime, )-&beta, d-glucopyranoside, 6-methoxykaempferol-3-O-&beta, d-glucopyranosyl-(6&prime, caffeoyl)-7-O-&beta, d-glucopyranoside, and 6,4&prime, dimethoxykaempferol-3-O-&beta, d-glucopyranoside. These results suggest that NZS exerts anti-obesity, anti-hyperlipidemia, and anti-hyperuricemic effects in HFD-induced obese mice, which are partly explained by regulation of lipid-metabolism-related genes and proteins and decreased expression of XOR.

Our previous report demonstrated successful isolation of soil-borne bacteria that suppressed the potential of Rhizoctonia solani AG2-2 (IV) causing turfgrass large patch disease when applied to Korean lawngrass ( Zoysia japonica). The current study aimed to uncover the mechanisms of this antagonism of Rhizoctonia solani and to define culture conditions for the isolated microbes. We found that two Bacillus isolates, I-009 and FRIN-001-1 strains, produced cellulase and siderophore, but not chitinase, while the Pseudomonas YPIN-022 strain was found to release only siderophore, implying that three antagonistic bacteria commonly interrupt Fe uptake by the large patch pathogen. The I-009 and FRIN-001-1 isolates grew best at 35 and 30°C in growth medium of pH 5 to 8 for 32 and 28 h, respectively, while optimum growth for the YPIN-022 strain was found at 35°C at pH 5 to 9 for 24 h. Good growth of I-009 and YPIN-022 over 24 h was obtained in M9 minimal medium supplemented with 1% sucrose, 0.5% yeast extract and 0.1% potassium chloride. FRIN-001-1 grew well in M9 medium with 1% mannitol, 0.5% yeast extract and 0.1% potassium phosphate dibasic.Additional key words

Camellia japonica is the most well-known species of the genus Camellia, which is native to Korea, Japan, and China. In this study, the antioxidant and free radical-scavenging activities of petal extracts of Camellia ecotypes with different colors were evaluated. For total phenolic compounds, five ecotypes were grouped into low [(4.8 mg·g−1 of dry weight (DW) GAE (gallic acid equivalent)], medium (6.0–6.2 mg·g−1) and high (19.6 mg·g−1) groups for white, red, and pink colors, respectively. The DPPH radical scavenging activity of the petal extracts, presented as a half maximal inhibitory concentration (IC50), was the greatest (3.8 μg·mL−1) in the pink ecotype and least (43.1 μg·mL−1) in the white ecotype as compared to ascorbic acid (13.6 μg·mL−1). The results demonstrated that the efficient DPPH radical scavenging activity of the pink ecotype was partly due to higher contents of phenolic compounds. Activities of two antioxidant enzymes, catalase and peroxidase, varied among the ecotypes, indicating the presence of ecotype-specific detoxifying processes. Camellia petals had higher amounts of antioxidants as well as excellent reactive oxygen species (ROS) scavenging activities that were dependent on the petal color.

This study was conducted to identify lily species native to Korea from formosan lily (Lilium formosanum) belonging to Longiflorum section. Due to flowering time, flower color and orientation, long shelf life and resistant to diseases, the native lily species can be valuable genetic resources for interspecific hybrids. One of the chloroplast genes, matK, was used to clone and sequence to explore any base changes. The matK was successfully amplified into 1,539 bp (94% of the gene) and phylogenetic tree demonstrated 6 clades for those 11 lily species used in this study. There were one or two base substitutions among 10 lilies native to Korea, while formosan lily native to Taiwan exhibited 6 base substitutions in matK gene, rendering it genetically distant. A restriction enzyme NruI recognized one of the six base changes, and digested the matK gene of 10 native lily species only, but not in formosan lily. The confirmed cleavage characteristic of the target region in matK gene was designed into a CAPS (cleaved amplified polymorphic sequences) marker which will be available to estimate compatibility of interspecific hybridization and to trace the pedigree when those native lilies are crossed with the formosan lily.

Cultivated watermelon (Citrullus lanatus) is one of the most important food crops in the Cucurbitaceae family. Diversification after domestication has led cultivated watermelons to exhibit diverse fruit flesh colors, including red, yellow, and orange. Recently, there has been increased interest in red-fleshed watermelons because they contain the antioxidant cis-isomeric lycopene. We performed whole genome resequencing (WGRS) of 24 watermelons with different flesh colors to identify single-nucleotide polymorphisms (SNPs) related to high lycopene content. The resequencing data revealed 203,894-279,412 SNPs from read mapping between inbred lines and the 97103 reference genome. In total, 295,065 filtered SNPs were identified, which had an average polymorphism information content of 0.297. Most of these SNPs were intergenic (90.1%) and possessed a transversion (Tv) rate of 31.64%. Overall, 2,369 SNPs were chosen at 0.5 Mb physical intervals to analyze genetic diversity across the 24 inbred lines. A neighbor-joining dendrogram and principal coordinate analysis (PCA) based on the 2,369 SNPs revealed that the 24 inbred lines could be grouped into high and low lycopene-type watermelons. In addition, we analyzed SNPs that could discriminate high lycopene content, red-fleshed watermelon from low lycopene, yellow or orange watermelon inbred lines. For validation, 19 SNPs (designated as WMHL1-19) were chosen randomly, and cleavage amplified polymorphic sequence (CAPS) markers were designed. Genotyping of the above 24 lines and 12 additional commercial cultivars using WMHL1-19 CAPS markers resulted in match rates of over 0.92 for most validated markers in correlation with the flesh color phenotypes. Our results provide valuable genomic information regarding the high lycopene content phenotype of red-fleshed cultivated watermelons, and the identified SNPs will be useful for the development of molecular markers in the marker-assisted breeding of watermelons with high lycopene content.

Two medium-leaf ecotypes (CY6069, CY6097) belonging to one species (Zoysia japonica) of Korean lawngrasses were selected in sod production fields in Jang Seong, Korea. They were reported to have distinct morphological and growth rate characteristics different from the preferred medium-leaf type zoysiagrass in Korea. This study was conducted to define further the genotypic difference at the molecular level and to develop DNA marker based on the specific DNA fragment. Polymorphic DNA fragments were first explored by using randomly amplified polymorphic DNA (RAPD) primers, which were then converted into PCR-based sequence characterized amplified region (SCAR) markers. The CY6069-specific primer set amplified about 550 bp successfully, while the CY6097 marker produced the expected 690 bp band, by which those markers were nominated by CY6069_550 and CY6069_690 SCARs, respectively. Together with the reported morphological and other phenotypic features, the SCAR markers confirmed in this study will be useful to identify those medium-leaf zoysiagrass genotypes when they are cultivated with other vegetatively propagated warm-season turfgrasses in sod farms.

This study investigated the effect of exogenous hydrogen peroxide (H₂O₂) on the antioxidant responses and growth of warm-season turfgrass (Zoysia japonica Steud.) and cool-season turfgrass (Poa pratensis L.) subjected to drought stress. Compared with control plants that were not pretreated with H₂O₂, plants pretreated with H₂O₂ had significantly greater fresh and dry weights of shoots and roots, and increased water content. H₂O₂ pretreatments before drought stress significantly decreased the concentrations of malondialdehyde and H₂O₂. DPPH radical scavenging and glutathione activities were significantly increased. The responsive activities of the antioxidant enzymes superoxide dismutase, ascorbate peroxidase, catalase, and peroxidase were also significantly enhanced. Our results suggest that exogenous H₂O₂ could improve the growth of warm-season and cool-season turfgrass under drought stress by increasing the activity of their antioxidant enzymes, while decreasing lipid peroxidation.

In order to evaluate effects of γ-rays on adventitious root formation and ginsenoside production, embryogenic calli induced from cotyledon explants of Panax ginseng C.A. Meyer were treated with γ-rays of 0, 10, 30, 50, 70, and 100 Gy. The highest frequency of adventitious root formation of 75 % occurred at γ-irradiation of 30 Gy, which is considered adequate dosage for selecting mutant cell lines. Five mutated adventitious roots (MAR)3-lines out of the propagation of 142 adventitious root lines treated with 30 Gy were selected based a 100-fold increase in proliferation rate compared to control adventitious roots (CAR) and content of the seven major ginsenosides (Rb1, Rb2, Rc, Rd, Re, Rf, and Rg1) was determined. In the CAR and four of the MAR3-lines (except for MAR3-109), the Rb/Rg ratio was greater than 1.0, thereby indicating altered ginsenoside composition in these root lines. The HPLC analysis of the MAR3-13 and MAR3-26 lines confirmed different ginsenoside profiles, including the three unidentified ginsenoside candidates, Gm1, Gm2, and Gm3. The ginsenosides of the MAR3-13 and MAR3-26 lines showed high hydroxyl and superoxide radical scavenging activities.

We investigated the structural genes and their transcripts for anthocyanin synthesis inDendranthema grandiflorum ‘Argus’. Color variations in chrysanthemum mutants were obtained through gamm ray irradiation to regenerated plants from anin vitro. Normal florets were pinkish, but the mutants had white or purple ray florets and white, purple, or yellow-green disc florets. Irradiation modified both flower size and the number of ray florets. Compared with the control, levels of total anthocyanins in the mutants ranged from 4 times lower to 6 times higher for the disc florets. This disparity was even more evident, up to 14-fold greater, in the ray florets. Expression of the CHI, F3′H, F3′5′H, DFR, and LDOX genes varied among the mutants, but no dramatic changes were detected in CHS and F3H transcripts in either leaf or floret tissues. Sequence homology to known anthocyanin genes from other plant species was 61 to 84%, 62 to 74%, and 71 to 76% for CHI, F3′H, and LDOX, respectively. Our results support the proposal that such radiation-induced mutations in genes within the anthocyanin pathway are associated with variations in chrysanthemum flower color.

Synthesis of organic compounds in response to salinity stress and their contribution to organic osmotic adjustment were investigated in seashore paspalum ( Paspalum vaginatum Swartz). Nine genotypes exhibiting the widest range of salt tolerance were grown in sea-salt amended nutrient solution in a greenhouse. Salinity ranges were 1.1 (EC w 0, control) to 49.7 dS m −1 (EC w 50) based on electrical conductivity of the solution (EC w ). Organic osmolytes most important within seashore paspalum under salinity stress were proline, Gly-betaine, and trigonelline in terms of explaining intraspecific salt tolerance differences and, therefore, should be the focus of biotechnology approaches to enhance these traits. While these osmolytes differed in accumulation with increasing salinity and absolute concentrations among salt tolerant and intolerant genotypes, the magnitude of responses was not sufficiently large to suggest use for salt screening as physiological/biochemical markers. Fructose concentration increased with salinity, especially for salt sensitive ecotypes, and may have potential as a marker. Glucose, sucrose, and myo -inositol tended to increase with salinity, but changes did not relate to intraspecific salt tolerance, while mannitol and sorbitol were not affected by salinity. Proline demonstrated a 20.8-fold increase averaged across genotypes from EC w 0 to EC w 50 salinity. Proline was the primary organic osmolyte for osmotic adjustment accounting for an average of 9.3% to total solute potential ( Ψ s ) at EC w 50 and 56% of the organic solute contribution to Ψ s . In the salt tolerant genotype, SI 93-2, proline and Gly-betaine exhibited greater absolute concentration and accumulation rate relative to the least salt tolerant, Adalayd. The intraspecific role of Gly-betaine did not relate to osmotic adjustment differences, suggesting another role perhaps in protection of the thylakoid membrane.

To develop rice (Oryza sativa L.) cultivars to be planted on salt-affected sites, cell lines with enhanced proline content and resistance to growth inhibition by Azetidine-2-carboxylic acid (AZCA), a proline analogue, were screened out among calli irradiated with gamma ray of 50, 70, 90, and 120 Gy. The calli had been derived from embryo culture of the cultivar Donganbyeo. Selected AZCA resistant lines that had high proline accumulation were used as sources for selection of NaCl resistant lines. To determine an optimum concentration for selection of NaCl resistant lines, Donganbyeo seeds were initially cultured on the media containing various NaCl concentrations (0 to 2.5%) for 40 days, and 1.5% NaCl concentration was determined as the optimum concentration. One hundred sixteen salt-tolerant (ST) lines were selected from bulked 20,000 seeds of the AZCA resistant seeds in the medium containing 1.5% NaCl. The putative 33 lines ( generation) considered with salt-tolerance were further analyzed for salt tolerance, amino acid and ion contents, and expression patterns of the salt tolerance-related genes. Out of the 33 lines, 7 lines were confirmed to have superior salt tolerance. Based on growth comparison of the entries, the selected mutant lines exhibited greater shoot length with average 1.5 times, root length with 1.3 times, root numbers with 1.1 times, and fresh weight with 1.5 times than control. Proline contents were increased maximum 20%, 100% and 20% in the leaf, seed and callus, respectively, of the selected lines. Compared to control, amino acid contents of the mutants were 24 to 29%, 49 to 143%, 32 to 60% higher in the leaf, seed and callus, respectively. The ratio of for most of the ST-lines were lower than that of control, ranging from 1.0 to 3.8 for the leaf and 11.5 to 28.5 for the root, while the control had 3.5 and 32.9 in the leaf and root, respectively. The transcription patterns for the P5CS and NHXI genes observed by RT-PCR analysis indicated that these genes were actively expressed under salt stress. The selected mutants will be useful for the development of rice cultivar resistant to salt stress.

Changes in photosynthetic capacity in variable salinity ranges were applied to explore mechanisms of salinity tolerance in seashore paspalum (Paspalum vaginatum Swartz) ecotypes. Nine ecotypes exhibiting a wide range of salinity tolerance were grown in a greenhouse using nutrient/sand culture, with six salinity levels of 1.1–49.7 dS m−1 (denoted as ECw0 to ECw50; electrical conductivity of water). With increasing salinity, chlorophyll concentrations decreased significantly only at ECw50 in comparison with nonsaline control. As salinity increased, initial chlorophyll fluorescence (F0) increased, while maximum fluorescence (Fm) and the variable to maximum fluorescence ratio (Fv/Fm) tended to decrease. As salinity increased, reflectance at visible wavelengths (507–706 nm) was enhanced, whereas it decreased at wavelength ≥760 nm. Compared to Adalayd, more tolerant SI 93-2 and HI 101 exhibited significantly lower reflectance in the photosynthetically active range (PAR), and higher reflectance beyond the PAR. All seashore paspalums maintained active photosynthetic capacity, as indicated by minor reductions in pigments, high light-harvesting capacity, and high maximum photochemical efficiency (high Fv/Fm values of 0.75–0.81) at high salt levels. SI 93-2 and HI 101 were characterized by high Fm and high Fv/Fm at ECw50 with minimal changes with increasing salinity. Also, SI 93-2 and HI 101 exhibited higher density and canopy cover. This resulted in low reflectance, as indicated by significantly higher IR/R (ratio of infrared to red wavelength) and NDVI (normalized difference vegetation index) indexes, and lower stress indexes (Stresses 1 and 2) compared to the least salt-tolerant Adalayd. IR/R and Stress 1 indexes were found to be useful tools for salinity tolerance evaluation, accounting for 85% of shoot and 51% of root growth variations, respectively.

Purpose: The current research reports antioxidant and free radical-scavenging activities of petal extracts from differently colored Camellia ecotypes. Methods: Five Camellia japonica ecotypes were chosen for petal extract. Total phenolic and flavonoid compounds were determined. For radical scavenging activities, DPPH andUWLC analysis were conducted. Results: For total phenolic compounds, five ecotypes showed the ranges of 4.8mg of GAE (gallic acid equivalent) perg dry weight (DW) to 19.6mg of GAE for white and pinkish petals, respectively. The DPPH radical scavenging activity of the petal extracts (represented in IC50)was highest (3.8 g·mL1) for the pinkish ecotype and lowest (43.1 g·mL-1) for the white ecotype when compared to the IC50 value for ascorbic acid (13.6 g·mL-1) as a positive control. The results demonstrate that the efficient DPPH radical scavenging activity of the pinkish ecotype was partly attributed to higher phenolic compounds. Activities of two antioxidant enzymes, catalase and peroxidase, were different among the ecotypes, indicating the presence of ecotype-specific detoxifying processes. Conclusion: The study demonstrates the potential use of the Camellia petals as an antioxidant resource, but there was a genotypic difference in total amount and antioxidant activities, indicating that more broad screening of the genotypes is necessary. Contact: Geung-Joo Lee, gjlee@cnu.ac.kr