Green Cultivars Research Articles

Light quality shapes morpho-functional traits and pigment content of green and red leaf cultivars of Atriplex hortensis

Recently the value of red-leaf species as food plants is increasing due to their high content in antioxidant compounds, mainly anthocyanins. Most work has been done on the modulation of light quality to maximise the production of antioxidant compounds in reddish leafy vegetables that mostly adjust the amount of foliar anthocyanins as a reaction to several environmental factors, including light. The aim of this study was to compare the effect of light quality on green- and red-leaf cultivars with a focus on the influence that different light wavelengths have on morpho-functional traits and pigment content. We selected as model plant Atriplex hortensis considering that cultivars with either fully-red or fully-green leaves are available. This species is characterized by the presence of betacyanins, an anthocyanin-homologue pigment known for its antioxidant properties. Plants were grown under four lighting treatments: 100% white light (W), 100% red light (R), red/blue light 50/50 % (RB), and red/green/blue light 33/33/33 % (RGB). All treatments provided a daily light integral (DLI) of 10.8 mol·m‒2·day‒1 over a 12-h photoperiod. Results showed that, in both green and red cultivars, light quality determines changes in morpho-functional traits and the combination of red and blue wavelengths enhances productivity and betacyanin content. In red plants, betacyanin content was two order of magnitude larger than in green plants, was significantly modulated by light quality, and increased according to the increasing percentage of blue wavelengths within the light spectrum. In the framework of enhancing antioxidant compounds in plant food through the adjustment of light spectrum, fully red plants should be considered as more promising than green cultivars.

Evaluation of bitter melon (Momordica charantia) cultivars grown in Texas and levels of various phytonutrients.

In the USA, Momordica charantia is relatively unknown and is usually found in specialty markets. In the present study, cultivation of five bitter melon cultivars grown under field conditions in College Station (TX, USA), was evaluated. Additionally, ascorbic acid, amino acids and phenolic compounds were quantified from various cultivars grown in different years. The yield of the first year of evaluation was comparable to other bitter melon growing regions, ranging from 9371.5kg ha-1 for the Japanese Spindle cultivar to 20 839.1kg ha-1 for the Hong Kong Green cultivar. Multivariate analysis suggests a strong correlation between yield and growth degree days, water use efficiency and organic matter, as well as an inverse correlation with the amount or precipitation during the growing season. The highest levels of total ascorbic acid were shown in the Japanese Spindle cultivar (162.97 mg 100 g-1 fresh fruit), whereas the lowest levels were expressed in the Hong Kong Green cultivar (42.69 mg 100 g-1 fresh fruit). The highest levels of total phenolics were consistently found the Indian White cultivar, in the range 10.6-12.5mg g-1 catechin equivalents. Seven phenolics and organic acids were identified and quantified by liquid chromatography-mass spectrometry and high-performance liquid chromatography, respectively. Additionally, the highest levels of total amino acids were found in the Large Top cultivar. The current 3-year field study demonstrates that it is feasible to grow bitter melon commercially in Texas with proper climatic and agronomic conditions. Bitter melon is a rich source for ascorbic acid, amino acids and phenolic compounds, which makes it a valuable food source with respect to improving human health. © 2018 Society of Chemical Industry.

Anthocyanins accumulation and molecular analysis of correlated genes by metabolome and transcriptome in green and purple asparaguses (Asparagus officinalis, L.)

Asparagus (A. officinalis L.) is a highly nutrition vegetable crop. Here, three purple asparagus cultivars, namely, Jing Zi-2, Purple Passion and Pacific Purple, and one green cultivar, namely, Jing Lv-1 were studied. At least 16 kinds of anthocyanins were identified in purple and green cultivars, and peonidin, cyanidin and their glycoside derivatives were found to be the major anthocyanins. Transcriptome data showed that most anthocyanin biosynthetic genes and at least 5 kinds of transcription factors were significantly differentially expressed significantly between the green and purple cultivars. Dark-treated experiments revealed that anthocyanins are not produced in the absence of light, and both the anthocyanin biosynthetic and regulatory genes were down-regulated greatly in the dark, implying that anthocyanins accumulation in asparagus is light-dependent. Overall, the results of this study provide useful information for understanding anthocyanin accumulation and the molecular mechanism of anthocyanin biosynthesis in asparagus.

Effect of decortication methods on the chemical composition, antinutrients, Ca, P and Fe contents of two pearl millet cultivars during storage

PurposeThe purpose of this paper is to evaluate the impact of modern and traditional decortication methods on the chemical composition, antinutrients, Ca, P and Fe contents during storage of two pearl millet cultivars (white and green) grown in Sudan.Design/methodology/approachThe grains of each cultivar were either decorticated traditionally using traditional stone dehuller or by using modern dehuller. The chemical composition, antinutrients and Ca, P and Fe contents of the cultivars were evaluated during storage for six months.FindingsThe results showed that both methods of decortication (modern and traditional) employed significantly decreased ash, protein, oil and crude fiber contents but increased moisture and nitrogen free extract contents of the grains for both cultivars. Storage resulted in a slight and gradual decrease in the chemical composition of the treated and untreated grains of the cultivars. Modern decortication of the grains significantly (P=0.05) reduced tannin content in both cultivars compared to untreated and traditionally decorticated grains. Phytic acid content of the white cultivar was not affected by the method of decortication used but modern decortication reduced that of green cultivar. Decrease in tannin and phytic acid was observed as the storage continued in both treated and untreated cultivars. Decortication significantly (P=0.05) reduced the Ca, P and Fe contents in both cultivars grains. Lower P and Fe contents were found in modern decorticated grains compared to traditional decorticated ones. Storage of the cultivars resulted in gradual decrease in Ca, P and Fe contents.Research limitations/implicationsProcessing methods such as decortication affect the quality attributes of pearl millet cultivars. The application of modern decortication method on pearl millet cultivars has better quality attributes than those treated with traditional decortication.Originality/valueThe study uses decortication methods (traditional and modern) to improve the quality attributes of pearl millet cultivars. Antinutrients such as tannin and phytic acid were observed to reduce the bioavailability of minerals like Fe. Decortication of the grains significantly reduced the level of such antinutrients and improved bioavailability of minerals.

Higher anthocyanin accumulation associated with higher transcription levels of anthocyanin biosynthesis genes in spinach.

Spinach (Spinacia oleracea L.) is widely cultivated as an economically important green leafy vegetable crop for fresh and processing consumption. The red-purple spinach shows abundant anthocyanin accumulation in the leaf and leaf petiole. However, the molecular mechanisms of anthocyanin synthesis in this species are still undetermined. In the present study, we investigated pigment formation and identified anthocyanin biosynthetic genes in spinach. We also analyzed the expression of these genes in purple and green cultivars by quantitative PCR. The accumulation of anthocyanin showed that it was the dominant pigment resulting in the red coloration in spinach. In total, 22 biosynthesis genes and 25 regulatory genes were identified in spinach, based on the spinach genomic and transcriptomic database. Furthermore, the expression patterns of genes encoding enzymes indicated that SoPAL, SoUFGT3, and SoUFGT4 were possible candidate genes for anthocyanin biosynthesis in red-purple spinach. The expression patterns of transcription factors indicated that two SoMYB genes, three SobHLH genes, and one SoWD40 gene were drastically up-regulated and co-expression in red-purple spinach, suggesting an essential role of regulatory genes in the anthocyanin biosynthesis of spinach. These results will enhance our understanding of the molecular mechanisms of anthocyanin biosynthesis in purple spinach.

Influence of soil and soilless agricultural growing system on postharvest quality of three ready-to-use multi-leaf lettuce cultivars

In this study the influence of soil and soilless growing systems (substrate 3:1 v/v perlite:peat) on quality and microbial traits of three multi-leaf lettuce cultivars (two green, ‘Eztoril’ and ‘Ezabel’, and one red, ‘Ezra’) was evaluated at harvest and after 7 and 13 days of storage at 8°C. At harvest, ‘Ezra’ showed a respiration activity and a total phenol content respectively 2-fold and 25% significantly higher than the green cultivars. Soil lettuces resulted more stressed than those grown in soilless, as indicated by their initial content in antioxidants. As for nitrate content, soilless grown lettuces at harvest showed an average concentration higher than soil-grown ones, although values are generally lower than limits imposed by the EU Regulation (No. 1258/2011). During storage, soilless lettuces showed no ammonium accumulation, differently from those cultivated in soil. In addition, lettuce cultivars grown in soilless condition showed unchanged content in the antioxidant activity and total phenols, and lower microbial counts than soil lettuces. Results of the present study showed that soilless growing system can positively affect qualitative and microbiological parameter of lettuces studied, and it can be considered a good soilless growing technique in order to obtain high quality multi-leaf lettuces for ready-to-use industry.

Evaluation of a New Mexico Landrace and Two Commercial Chile (Capsicum annuum) Cultivars under Four Furrow Irrigation Schedules

Commercial and landrace chile (Capsicum annuum) cultivars are cultivated under furrow irrigation systems in Northern New Mexico. Yield and physiological differences between commercial and landrace chile cultivars under furrow irrigation systems have not been evaluated. In 2011 and 2012 two commercial chiles, ‘Sandia’ and ‘NuMex Big Jim’, with one landrace chile, ‘Chimayo’, were evaluated under four irrigation schedules, with irrigation once every 7, 9, 11, and 13-days. These four schedules represent possible water availability for farmers in Northern New Mexico. In 2011 there were inconsistent yield patterns; fresh red chile yield of ‘Chimayo’ at the seven-day interval was 90% more than at the nine-day interval. ‘Sandia’ had 138% better yields at the seven- than at the nine-day interval. ‘Chimayo’ fresh green chile yields at the nine-day interval were 47% better than the seven-day interval. ‘NuMex Big Jim’ fresh green yields were 40% greater at the seven-day interval than the 13-day interval. In 2012 no yield components were statistically different for cultivars across irrigation intervals. This data shows commercial green and landrace chile cultivars can be furrow irrigated as water becomes available on 7, 9, 11, or 13-day intervals with no yield effect.

Comprehensive Dissection of Metabolic Changes in Albino and Green Tea Cultivars.

Albino tea cultivars are special mutants of tea plants with white or yellow leaf color. In this study, three albino tea cultivars, including 'Anji Baicha', 'Huangjinya', and 'Baijiguan', and two green tea cultivars, 'Longjing 43' and 'Fuding Dabaicha', were applied to metabolite profiling by gas chromatography-mass spectrometry and ultraperformance liquid chromatography-mass spectrometry. Multivariate analyses revealed significantly different metabolite phenotypes in leaves among albino cultivars and green cultivars. The differential metabolite-related pathways included galactose metabolism, tryptophan metabolism, phenylpropanoid biosynthesis, and flavonoid biosynthesis. For the young leaves of albino cultivars, the sugar (sorbitol and erythrose) and amino acid (mainly proline, isoleucine, ornithine, aspartic acid, threonine, and valine) concentrations increased, whereas gallocatechin and epigallocatechin gallate concentrations decreased. These results reveal the divergence in metabolic profiling between tea plant cultivars with different leaf colors. With the development of leaves, the concentrations of flavonoids increased largely in the older leaves of albino cultivars.

Ineffectiveness of foliar S-ABA application as an apple sunburn suppressant explained through effects on peel biochemistry and leaf ecophysiology

Abstract Sunburn is a physiological disorder that affects the visual quality of apple (Malus Domestica) fruit. In South Africa, producers estimate yield losses of up to 50% in green cultivars due to sunburn damage. Recent research in Japan has found that foliar application of abscisic acid (S-ABA) reduced sunburn in apples by up to 30%. The aim of this study was to examine the effect of S-ABA application on the occurrence of sunburn and on other fruit quality parameters in ‘Granny Smith’, the apple cultivar that suffers most sunburn under South African conditions. Trials were conducted over three growing seasons from 2010–2011 to 2012–2013 in a ‘Granny Smith’ orchard in Grabouw, South Africa. S-ABA was applied at concentrations between 250 and 1000 ppm and various timings during summer (from November until harvest in March/April). A representative scaffold branch on both sides of the tree was strip picked at commercial harvest resulting in samples of at least 100 fruit per tree for sunburn, fruit color, and red blush assessment. A sub-sample of 20 fruit was randomly selected and used to determine average fruit size, fruit firmness, and internal quality. The application of S-ABA did not decrease sunburn incidence and severity under South African growing conditions. S-ABA application decreased the peel concentration of total antioxidants, total phenolics and reduced ascorbic acid whilst increasing oxidized ascorbic acid. These changes suggest that S-ABA either downregulated synthesis of antioxidants or caused increased oxidative stress. Consistent with S-ABA application possibly causing stress, leaf necrosis was observed when S-ABA was applied just prior to periods of high temperature. Consistent with its physiological role in plants, S-ABA application decreased stomatal conductance and thereby also decreased the net carbon assimilation and transpiration rates, while the stem water potential was increased due to reduced water loss. Concomitant with the decrease in carbon assimilation, there was a significant reduction in total soluble solids, and titratable acidity with repeated S-ABA applications. S-ABA did not affect fruit maturity. Our results suggest that application of S-ABA to reduce sunburn is ineffective under South African conditions and therefore not recommended.

Blue and Red LED Illumination Improves Growth and Bioactive Compounds Contents in Acyanic and Cyanic Ocimum basilicum L. Microgreens.

Microgreens are an excellent source of health-maintaining compounds, and the accumulation of these compounds in plant tissues may be stimulated by exogenous stimuli. While light quality effects on green basil microgreens are known, the present paper aims at improving the quality of acyanic (green) and cyanic (red) basil microgreens with different ratios of LED blue and red illumination. Growth, assimilatory and anthocyanin pigments, chlorophyll fluorescence, total phenolic, flavonoids, selected phenolic acid contents and antioxidant activity were assessed in microgreens grown for 17 days. Growth of microgreens was enhanced with predominantly blue illumination, larger cotyledon area and higher fresh mass. The same treatment elevated chlorophyll a and anthocyanin pigments contents. Colored light treatments decreased chlorophyll fluorescence ΦPSII values significantly in the green cultivar. Stimulation of phenolic synthesis and free radical scavenging activity were improved by predominantly red light in the green cultivar (up to 1.87 fold) and by predominantly blue light in the red cultivar (up to 1.73 fold). Rosmarinic and gallic acid synthesis was higher (up to 15- and 4-fold, respectively, compared to white treatment) in predominantly blue illumination. Red and blue LED ratios can be tailored to induce superior growth and phenolic contents in both red and green basil microgreens, as a convenient tool for producing higher quality foods.

Antioxidant Activity of the Phenolic and Oily Fractions of Different Sweet Bell Peppers

The content of phenols and ascorbic acid of the phenolic fraction, and carotenoids, tocopherols and capsaicinoids of the oily fraction from sweet bell peppers from northwest Mexico was determined. Antioxidant activity in both fractions was evaluated (ABTS and DPPH methods). Green cultivar had the highest content of phenols, flavonoids, and ascorbic acid and highest antioxidant activity. α-Tocopherolwas found in the four cultivars; however, capsaicinoids were not detected. The phenolic fraction had higher antioxidant activity than the oily fraction.

Identification and Characterization of Anthocyanin Biosynthesis-Related Genes in Kohlrabi.

Kohlrabi (Brassica oleracea var. gongylodes L.) is an important vegetable of the Brassicaceae family. The main edible part of kohlrabi is the swollen stem. The purple cultivars make anthocyanin mainly in the peel of the swollen stem, while in the leaf, it is limited to the midrib, but green cultivars do not. Anthocyanins are advantageous for both plants as well as humans. Two anthocyanin compounds were detected by high pressure liquid chromatography (HPLC) only in the peel of the purple kohlrabi cultivar. Three MYBs, three bHLHs, and one WD40 TF were identified as the candidate regulatory genes in kohlrabi. There was an abundance of transcript levels for the late biosynthetic genes more specifically for BoF3'H, BoDFR, BoLDOX, and BoGST in the purple peel while scarcely detectable in other tissues for both cultivars. The expression of BoPAP2 and BoTT8 was higher in the peel of the purple cultivar than the green cultivar. The expression of BoMYBL2.2 orthologue of Arabidopsis MYBL2, a negative regulator of anthocyanins, was dramatically decreased in the purple peel. The expression of BoACO1, a key gene for ethylene biosynthesis, and BoNCED3, an important gene of the ABA pathway, was down- and upregulated, respectively, in the peel of purple kohlrabi.

Ascorbic Acid, Total Phenolic, Flavonoid and Antioxidant Activity of Two Cultivars of Basella alba

Antioxidants and secondary metabolites have attracted a great deal of attention for their effect in preventing disease due to oxidative stress. In this study, two cultivars of Basella alba were evaluated for the ascorbic acid, total phenolic, flavonoid and total antioxidant activities by standard methods. Ascorbic acid of green cultivar was 19.38 mg/100g and red cultivar was 25.85 mg/100g fresh weight, highly significant difference (p < 0.05) was observed in ascorbic acid. Total phenolic contents were 61.00 and 90.52 mg/g fresh weight for the green and red cultivar respectively, flavonoid content of the green cultivar was 13.37 mg/g while red cultivar was 17.7mg/g significant difference was observed in the ferric reducing power of the two cultivars. DPPH of green cultivar was 78.85% while for red cultivar was 79.81%, no significant difference was observed between the cultivars. Total antioxidant activity for green cultivar was 60.86 mg/g while for red cultivar was 70.38 mg/g fresh weight, no significant difference was observed in the total antioxidant activity. Both cultivars were excellent sources of ascorbic acid, total phenolic compound, flavonoid and total antioxidant properties although red cultivar had higher values.

Developmental and hormonal regulation of fiber quality in two natural-colored cotton cultivars

AbstractCotton cultivars with brown (Xiangcaimian 2), green (Wanmian 39) and white (Sumian 9) fiber were investigated to study fiber developmental characteristics of natural-colored cotton and the effect of hormones on fiber quality at different stages after anthesis. Fiber lengths of both natural-colored cottons were lower than the white-fibered control, with brown-fibered cotton longer than green. Fiber strength, micronaire and maturation of natural-colored cotton were also lower than the control. The shorter fiber of the green cultivar was due to slower growth during 10 to 30 days post-anthesis (DPA). Likewise, the lower fiber strength, micronaire and maturation of natured-colored cotton were also due to slower growth during this pivotal stage. Indole-3-acetic acid (IAA) content at 10 DPA, and abscisic acid (ABA) content at 30 to 40 DPA were lower in the fibers of the natural-colored than that of the white-fibered cotton. After applying 20 mg L−1 gibberellic acid (GA3), the IAA content at 20 DPA in the brown and green-fibered cottons increased by 51.07 and 64.33%, fiber ABA content increased by 38.96 and 24.40%, and fiber length increased by 8.13 and 13.96%, respectively. Fiber strength, micronaire and maturation were also enhanced at boll opening stage. Those results suggest that the level of endogenous hormones affect fiber quality. Application of external hormones can increase hormone content in natural-colored cotton fiber, improving its quality.

Effect of seed sowing period on polyphenolic compounds content in basil (Ocimum basilicum L.) under greenhouse conditions

The object of the research is the process of formation of the polyphenolic complex by basil plants. The complexity in evaluating the process of polyphenols accumulation lies in the fact that the formation of these compounds occurs under the influence of a number of biotic and abiotic factors, leading to variation in the level of polyphenols in a very wide range even for plants of the same cultivar.The influence of different seed sowing periods of five cultivars of basil on the accumulation of polyphenolic compounds in each cutting of green mass was investigated. The obtained data were mathematically computed using AgrostatNew and MicrosoftExcel software. It was found that cultivars with purple coloration of leaves accumulate by 51.2–66.6 % more polyphenols than green cultivars. It was observed that in the green cultivars Bad’oryi, Rutan and Siaivo, the level of polyphenols increases with each subsequent cutting of green mass during the February and April seed sowing periods. The cultivars with purple coloration of leaves have shown the highest level of accumulated polyphenols in the first cutting for the February seed sowing period and in the fourth and fifth cutting in April. The level of polyphenols in all the cultivars, which were sown in March, did not significantly change during cutting, indicating the optimal cultivation conditions.

AAC Oriole red field pea

AAC Oriole is a semi-leafless red field pea (Pisum sativum L.) cultivar developed at the Lacombe Research and Development Centre, Agriculture and Agri-Food Canada, Lacombe, AB. It is shorter than most of the recently released yellow and green pea cultivars on the Canadian market with fair standability. It has early maturity, medium seed size, and round seed shape with a crude seed protein content of approximately 24%. AAC Oriole is resistant to powdery mildew caused by Erysiphe pisi DC. and moderately susceptible to mycosphaerella blight [caused by Mycosphaerella pinodes (Berk. & A. Blox.) Vestergr.] and fusarium wilt [caused by Fusarium oxysporum f. pisi (Linford) W.C. Snyder & H.N. Hansen]. It is adapted to all field pea growing regions in western Canada.

Spectral dependence of electrical energy-based photosynthetic efficiency at single leaf and canopy levels in green- and red-leaf lettuces

The spectrum of light affects both the electrical energy consumption by plants and photosynthetic efficiency. In a plant factory, where light-emitting diodes (LEDs) serve as an alternative to solar light, the optimal spectrum of light should be carefully chosen to maximize the rate of photosynthesis and the electrical energy efficiency of the crop. The objectives of this study were to investigate the photosynthetic rate of different colored lettuces (reddish and green leaves), to quantify the spectral dependence of photosynthetic efficiency, and to optimize the LED spectrum for maximum canopy photosynthesis and electrical energy consumption in lettuce grown in a plant factory. Two lettuce cultivars (Lactuca sativa L.), ‘JeokChukMyeon’ and ‘CheongChukMyeon’, were assessed for light absorption and photosynthetic efficiency at the single leaf and canopy levels, and the relative consumption of electrical energy from the LED lights was measured at 18 narrow wavelength bands of 10 nm from 400 to 700 nm. Anthocyanin and chlorophyll content (SPAD value) were measured and correlated with leaf color. Light interception by the canopy was estimated with light transmittance models. The light absorption was similar among the green and reddish lettuce cultivars at most wavelengths, but slightly higher in the reddish leaves around 550 nm (green region). In the reddish leaves, photosynthetic rates per incident photon of a single leaf had two peaks at 650-660 and 400-410 nm, while the photosynthetic rate per absorbed photon had three peaks at 650-660, 400-410, and 540-560 nm. In the green region of the light spectrum, both photosynthetic rates per incident photon and those per absorbed photon were lower in the reddish cultivars than in the green cultivars. The spectral dependence of light absorption at the canopy level was much weaker than that at the single leaf level. The quantum yield and absorption of green light at the canopy level were nearly same as those of blue and red lights, indicating that the photosynthetic efficiency of green light at the canopy level was higher than that at the single leaf level. The relative electrical energy consumption was lower in the green region than in the red and blue regions. Therefore, the photosynthetic efficiency based on electrical energy consumption at the canopy level was much lower with green LEDs than with blue or red LEDs. These results describe the plant response to the light spectrum at the canopy level and can be useful for optimizing artificial lighting sources for maximum plant productivity and energy-savings in a plant factory.

Performance of irrigated green corn cultivars in different plant populations at sowing

ABSTRACT This study aimed to evaluate the yield of green corn hybrids grown under irrigation in different plant populations at sowing. The assay was carried out in the experimental area located in the city of Arapiraca, Alagoas State, Brazil, from November 2015 to January 2016. A randomized complete block design (RCBD) was used, in a 2 x 5 factorial scheme with four replicates. A double- and a single-cross hybrid (AG 1051 and BM 3061), which are suitable for green corn production, were cultivated in five spacings between plants at sowing (15.0, 17.5, 20.0, 22.5 and 25.0 cm). The characteristics photosynthetic rate (PR), ear length with rusk (HEL) and without husk (UEL), husked ear weight (HEW), unhusked ear weight (UEW), percentage of marketable ears (%ME) and weight of husk (HW) were evaluated. The double- and single-cross hybrids AG 1051 and BM 3061 showed green ears with commercial standard. The cultivar BM 3061 showed the best results for most of the characteristics related to the performance of green corn (PR, HEL, UEL, UEW, HEW, %ME). The spacing of 17.5 cm between plants at sowing was the most indicated for irrigated green corn cultivation.

Phylogenetic Relationship of Indonesian Water Yam (Dioscorea alata L.) Cultivars Based on DNA Marker Using ITS-rDNA Analysis

Research on genetic diversity and intra-species classification of Indonesian Dioscorea alata L. based on morphological characters has been done, and the result shows that there are 4 sub-groups of green cultivars group, and 5 sub-groups of purplish-red cultivars group. The objectives of this study are to determine the phylogenetic relationship cultivars of D. alata Indonesia compared to D. bulbifera as nearest species as well as 3 cultivars from GenBank. The young leaves of 18 water yam cultivar accessions were collected from Java, Madura, South Sumatera, South Kalimantan, Centre Celebes (Sulawesi), Ternate, West Papua, and Nusa Tenggara islands of Indonesia. DNA Isolation was conducted using Phytopure reagent. DNA amplification was conducted using thermocycler, predenaturation at 95 oC 5 minutes, denaturation at 95 oC 1 minutes, annealing at 60 oC 3 minutes, and elongation at 72 oC 2 minutes along with 30 cycles. The PCR products were electrophoresed in 1.5% agarose and visualized under UV transiluminator. Fifty-five µl of PCR products with positive targeted band between 700-800 bp were sent to 1st Base Singapore for purification and sequencing of 18S, ITS-1, 5.8S, ITS-2, and 28S rDNA. The DNA sequences were compared and aligned by BioEdit program (version 7.0.5.2) and MEGA programs. Comparison of entire sequences of the tested samples were aligned by software ClustalW (version 1.83). Phylogenetic trees were based on hierarchical clustering of the alignments of 18S, ITS-1, 5.8S, ITS-2, and 28S rDNA and produced by Neigbor-Joining using MEGA 5 software of the bootstrap values (1000 replicates). The study result shows that D. alata cultivars have high genetic variability on ITS-1, 5.8S, and ITS-2 rDNA region. Groups of green and purplish red cultivars formed based on morphological characters are not formed based on ITS-rDNA markers. Sub-groups were formed based on ITS-rDNA molecular markers derived from both the green and purplish-red cultivar groups. This result revealed that two cultivar groups are not similar with RAPD and morphological characters.

Active Volatile Constituents in Perilla frutescens Essential Oils and Improvement of Antimicrobial and Anti-Inflammatory Bioactivity by Fractionation

Perilla frutescens (L.) Britt Labiatae (Pl), known as “Zi-So” and widely grown in China (red cultivar, Rpl) and Japan (green cultivar, Gpl), is frequently served on “Sa-Shi-Mi” dishes. Considering whether the perilla leaves substantially possess antimicrobial activity, we prepared the essential oils (EO: EOr and EOg), the pentane and ether [E: E(rp) and E(gp)] subfractions, the aqueous [Rpl(w) and Gpl(w)] and ethanolic extracts [pl(E), Rpl(E) and Gpl(E)] to compare their biological activities. Results indicated Gpl(E) and Rpl(E) contained huge amount (in mg/100 g) of polyphenoilcs (1472±1.0 vs. 1269.8±1.7), rutin (20.8±0.8 vs. 11.4±0.6), quercetin (29.8±0.9 vs. 13.6±0.6), and lutein (13.9±0.7 vs. 14.2±0.8). The total number of volatile constituents present in EOr and EOg were 27 in each but different phytochemicals. The top five major constituents (in %) in EOr were perilla aldehyde (54.35%), limonene (23.81%), trans-caryophyllene (7.2%), cis,trans-α-farnescene (7.02%), and linalool (2.40%); that in EOg were perilla aldehyde (65.26%), limonene (12.49%), cis,trans-α-farnescene (7.31%), trans- caryophyllene (5.91%), and linalool (2.75%); The top two constituents in E(rp) were perilla aldehyde (93.19%) and linalool (4.02%); that of E(gp) were perilla aldehyde (84.40%) and linalool (5.03%). EO and E showed moderate DPPH free radical scavenging-, ferrous ion chelating-, superoxide anion- and H2O2 -scavenging capabilities. At 1.0 mg/mL the Es always revealed to be superior to EOs against E. coli, S. aureus, V. parahemolyticus, and T. mentagrophytes. Amazingly, E was stronger than perilla aldehyde against S. typhimurium,. The cytotoxicity test indicated Gpl(E) was more toxic than Rpl(E) and the pl(E)s were more toxic than pl(w)s. Conversely, the pl(w)s showed stronger TNF-α suppressing activity than pl(E)s. The pentane subfractions were entirely ineffective. Conclusively, the Perilla EOs contain huge amount of polyphenoilcs, the Es exhibit the rather good antimicrobial and cytotoxic effects, while the pl(w)s possess rather promising anti-inflammatory effect, underlying the promising antimicrobial and antiinflammtory effects of perilla leaves.