The extracts of Pinus densiflora sawdust by n hexane, ethyl acetate and methanol solvent were investigated to identify their mycelial growth inhibition against Lentinus edodes. The yields of n hexane soluble fraction, ethyl acetate-soluble fraction, and methanol soluble fraction from P. densiflora sawdust were obtained 1.36%, 2.21% and 4.03% using organic solvent, respectively. The mycelial growth inhibition of L. edodes was the greatest for n hexane extract, ranging from 36.5% to 47.6% at concentrations of 125 ppm to 1,000 ppm, with the values for all concentrations significantly different from one another. After direct extraction of P. densiflora sawdust using n hexane, ethyl acetate and methanol, each extract was separated into three fractions by silica gel column chromatography and then the fractions were isolated on the values of Rf by thin layer chromatography. The mycelial growth inhibition against L. edodes was recognized in the fractions II (33.5%) and III (37.6%) of n hexane extract, the fraction II (21.4%) of ethyl acetate extract and the fraction II (26.4%) of methanol extract. The fractions III of n-hexane extract showed the highest growth inhibition among the nine fractions of the organic solvent extract.
A survey was conducted during April~May 2004 to determine the occurrence and population density of plant-parasitic nematodes in ginseng (Panax ginseng C.A. Meyer) growing fields, in major ginseng growing regions of Chungbuk, Chungnam, Gyeongbuk and Kyongki provinces. The survey revealed presence of eleven species of plant-parasitic nematodes namely, Criconemoides morgensis, Ditylenchus destructor, Helicotylenchus dihystera, Meloidogyne incognita, M. hapla, Paratylencgus lepidus, Pratylenchus penetrans, Psilenchus hilarulus, Trichodorus similis, Tylenchorhynchus claytoni and Xiphinema americanum. Frequency and density of each species were highly variable. M. incognita and M. hapla were the predominant species, their infestation observed in 46.3 and 39.4% fields with an average density of 78~254 and 76~211 nematodes per 300 cm3 soil, respectively. Whereas, T. similis and X. americanum were rarely observed; only in 2.3 and 1.8% of surveyed fields and their density was 10~17 and 7~10 individuals per 300 cm3 soil, respectively. They are recorded herewith for the first time from ginseng fields of Korea. In nematode-infestated fields, stunted plant growth with chlorotic leaves, and wilted plants were observed in patches.
Rehmannia glutinosa shows a high level of resistance to the non-selective herbicide paraquat. To characterize the antioxidant enzyme system of R. glutinosa, we comparatively examined the responses of antioxidant enzymes to UV, wounding and a general elicitor yeast extract in R. glutinosa and soybean. The levels of enzyme activities of the two plant species were drastically different between those per fresh weight (general activity) and per protein (specific activity) bases. The general activities of superoxide dismutase (SOD), peroxidase (POX), catalase (CAT), and glutathione reductase (GR) were lower, but that of ascorbate peroxidase (APX) was higher in R. glutinosa than in soybean. The specific activities of the enzymes, however, were about two- to seven-fold higher in R. glutinosa than in soybean, except that of CAT, which was about 12-fold higher in soybean. The general and specific enzyme activities of R. glutinosa relative to those of soybean showed a consistent increase in responses to the stresses only in SOD. The specific activities of SOD and APX were higher in R. glutinosa in all stress treatments. The results might suggest a relatively higher contribution of SOD and APX to the stress tolerance.
Aconitine alkaloids produced from cell suspension cultures of Aconitum napellus for the first time. The effects of various culture conditions on cell biomass and aconitine accumulation in cell suspension cultures were investigated. Suspension cell cultures of A. napellus were established by transferring callus tissues from leaf explants onto liquid MS medium supplemented with 1 mg/l NAA and 0.1 mg/l kinetin. Among the culture media tested, MS medium had a pronounced effect on cell growth and aconitine accumulation. The maximum dry cell weight was obtained at inoculum size of 3 g (FCW) per flask and in MS medium supplemented with 5% sucrose after 8 weeks. The addition of salicylic acid (SA) and yeast extract (YE) in the MS medium enhanced aconitine accumulation. Using a proper combination of culture condition and supplements, aconitine content could reach 0.043% (dry weight basis), that was 2.5~3 fold higher that detected in control cultures.
Scopolia parviflora Nakai, a rare and endangered species, is the sole plant producing tropane alkaloids (TA) among the Korean native species. In order to enhance TA productivity the SP72 root line was selected by screening 100 of root line, and the optimal culture media for root growth and TA production were investigated with the SP72 roots. Based on the several media, SH and 2B5 medium were determined as growth medium and White and NN medium as production medium. Among the four combinations of two-stage culture, 2BN (2B5 as growth medium plus NN as production medium) showed more enhanced root growth and TA production as compared with production media of White and NN medium and growth media of SH and 2B5 medium, respectively. However, bubble column bioreactor (BCB) cultures applying two-stage culture did not reveal the effective results despite of the each successful operation of two-stage culture in conical flasks and BCB cultures.
Scopolamine and hyoscyamine are important anticholinergic compounds. To increase the productivity, we have selected various elicitors and developed culture system using a bubble column bioreactor (BCB). As the same manner of elicitation in flask cultures, the elicitors were introduced into BCB cultures and the productivity was investigated. Except the bacterial elicitor of Staphyllococcus aureus, the elicitors inhibited hyoscyamine production. In scopolamine production, the elicitors revealed different responses from the results obtained in flask cultures. The elicitors of KCl and Candida albicans less increased the production than flask cultures. However, methyl jasmonate and S. aureus showed stronger positive effects on tropane alkaloid production. In particular, S. aureus was the most effective elicitor on scopolamine production and the elicitor resulted in the highly increased production, approximately 10 times higher than the control culture.
An in vtro nucellar polyembryo propagation method was established with mature seed of the Citrus junos Sieb. 7-8 nucellar polyembryos per seed were induced on MS basal medium without plant growth regulators. The polyembryos developed to complete plantlets on teatment with IBA. These shoots grew further in MS medium without plant growth regulators. Rooting of shoots occurred on MS medium supplemented with IBA. These plantlets were successfully transplanted to small plastic pot containing soil mixture. Somatic embryos were induced from nucellar polyembryo and maturation occurred spontaneously from proliferating cultures on MS medium without growth regulators. Random Amplified Polymorphic DNA (RAPD) marker analysis of in vitro and in vivo grown junos orange showed identical polymorphism indicative of their genetic stability. The RAPD polymorphism produced revealed same banding pattern in each regenerant. Hence, propagaton of junos orange by nucellalr polyembryos was efficient and produced in genetically stable plants under in vitro conditions.
Optimal culture conditions for efficient in vitro propagation and polysaccharide production of Orostachys japonicus were established. O. japonicus was cultured in media containing various growth regulators and carbon sources. The highest regeneration rate was achieved in 1.0 and 3.0 mg l -1 of 2,4-D concentration, while the lowest was obtained in 10.0 mg l -1 2,4-D concentration. When different carbone sources were added in the culture medium, plant growth was high in 3% sucrose treatment. The micropropagated shoots were successfully acclimatized in artificial soils and produced comparable amont of polysaccharide compred to parent cultivated plants.
The chromosome numbers and karyotypes were investigated in four Korean native species of the genus Hosta. The chromosome complements were diploid of 2n=60 in H. japonica var. lancifolia Nakai and H. capitata Nakai, aneuploid of 2n=59 in H. minor (Bak.) Nakai, and modified triploid of 2n=92 in H. longipes (Fr. et Sav.) Matsumura. All the species carried four sets of distinctly large chromosomes of which the chromosome types were telocentrics or subtelocentrics with 4.4~7.2 μm in length. The other chromosomes were meta-, submeta, subtelo-, or telocentric types and showed gradual length degradation in the range of 1.0~3.0 μm. The satellites appeared vestigially in a pair or a triplet set of chromosomes which depends on the species. New chromosome number and karyotype in H. longipes were the first report in this species. The structural rearrangement was suggested to explain the modified triploid composition of 2n=92.
Karyotypes were established in the eight Korean native species of the genus Iris. Chromosome numbers were 2n=50 in I. koreana and 2n=42 in I. uniflora var. carinata and their karyotype formulas were K = 2n = 50 = 14m + 28sm + 8st and K = 2n = 42 = 16m + 26sm, respectively. I. dichotoma and I. pseudoacorus were diploids of 2n=34. However, they showed different karyotype formulas: K = 2n = 34 = 26m + 6sm + 2st in I. dichotoma and K = 2n = 34 = 8m + 24sm + 2st in I. pseudoacorus. I. setosa, and I. pallasii var. chinensis carried the same chromosome numbers of 2n=40, but they showed different patterns of karyotype formula: K = 2n = 40 = 22m + 14sm + 4st in I. setosa and K = 2n = 40 = 26m + 12sm + 2st in I. pallasii var. chinensis. I. sanguinea was a diploid of 2n=28 and the karyotype formula was K = 2n = 28 = 14m + 14sm. I. ensata var. spontanea was a diploid of 2n=24 and the karyotype formula was K = 2n = 24 = 10m + 14sm. Each species showed characteristic chromosome composition with a pair of satellite chromosome except I. koreana with three pairs of satellite chromosomes. The chromosomes of I. dichotoma and I. uniflora were comparatively short, while the chromosomes of I. ensata were remarkably bigger than those of other species. These cytological data will give a useful information for the identification and breeding program of the Iris plants.
Resveratrol, which is both a phytoalexin with antifungal activity and a phytochemical associated with reduced cancer risk and reduced cardiovascular disease, is synthesized in a limited number of plant species including peanut. Resveratrol synthesis is catalyzed by the enzyme stilbene synthase including resveratrol synthase (RS). Resveratrol synthase gene (RS3) obtained from peanut, Arachis hypogaea, Fabaceae has been transferred into chinese foxglove, Rehmannia glutinosa by using Agrobacterium mediated transformation. RS t-DNA introduced to chinese foxglove (R. glutinosa L) by transformation and its reaction product, resveratrol-3-O-β-D-glucoside was isolated and characterized using HPLC. Also its biological effects was tested in inhibition of the lipid peroxidation of mouse LDL by glycosylated stilbenes derivatives obtained from transgenic plants. Resveratrol-3-O-β-D-glucoside isolated from transgenic R. glutinosa L. showed antimicrobial activity of the growth inhibition zone against Escherichia coli and Salmonella typhimurium. Therefore, this compound can be contributed to be useful as a phytoalexin for plant health as well as a phytochemical for human health.
Myristica fragrans seed from Myristica fragrans Houtt (Myristicaceae) has various pharmacological activities peripherally and centrally. The present study aims to investigate the effect of the methanol extract of Myristica fragrans seed (MF) on kainic acid (KA)-induced neurotoxicity in primary cultured rat cerebellar granule neuron. MF, over a concentration range of 0.05 to 5 μg/ml inhibited KA (500 μM)-induced neuronal cell death, which was measured by trypan blue exclusion test and 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl-tetrazolium bromide (MTT) assay. MF (0.5 mug/ml) inhibited glutamate release into medium induced by KA (500 μM), which was measured by HPLC. Pretreatment of MF (0.5 mug/ml) inhibited KA (500 μM)-induced elevation of cytosolic calcium concentration ([Ca2+]c), which was measured by a fluorescent dye, Fura 2-AM, and generation of reactive oxygen species (ROS). These results suggest that MF prevents KA-induced neuronal cell damage in vitro.
Microsomal Ω-3 fatty acid desaturase (FAD3) is an essential enzyme in the production of the n-3 polyunsaturated fatty acid α-linolenic acid during the seed developing stage. To understand the regulatory mechanism of the gene encoding the Ω-3 fatty acid desaturase, a genomic fragment corresponding to the previously isolated perilla seed PfFAD3 cDNA was amplified from perilla (Perilla frutescens Britt) by GenomeWalker PCR. Sequence analysis of the fragment provided with identification of a 1485-bp 5'-upstream region and a 241-bp intron in the open reading frame. To determine the tissue-specificity of the PfFAD3 gene expression, the 5'-upstream region was fused to the β-glucuronidase (GUS) gene and incorporated into Arabidopsis thaliana. Histochemical assay of the transgenic plants showed that GUS expression was restricted to seed and pollen, showing that PfFAD3 gene was exclusively expressed in those tissues.
Alkaloid producing species of plants have long been a major component of the medicinal social and magico-religious aspects of human culture. A diverse array of biological activities has been attributed to different alkaloids including numerous members of benzylisoquinoline family of alkaloids. For biotechnological approaches of alkaloid biosynthesis in poppy family, plant regeneration protocol through somatic embryogenesis or shoot organogenesis is a first step. This paper describes the methods and applications of plant regeneration of poppy family.