Pine wilt disease (PWD) induced by the pinewood nematode (PWN), Bursaphelenchus xylophilus, is a great threat to pine forests in Europe and East Asia. Identification of interrelation between Monochamus spp., the major vectors of PWD and PWN is the key factor for effective control of PWD because PWN is trasmitted to healthy pine trees only by Monochamus vectors. In this study, direct injection system is designed to apply PWN artificially to M. alternatus just before emergence to adults. After injection of PWN (300~500 nematodes in 10 μl of 1X PBS) using adapted glass pipets, the survival rate of M. alternatus and detection rate of live PWN were 80.0% and 68.8% at 168 h respectively. As a result, we demonstrated the artificial injection methods of nematode to Monochamus vectors for further study of coeffect between vectors and specific stage of PWN including 3rd stage dispersal juvenile.

Non-target predatory insects can be indirectly exposed to aerial pesticide spraying and fogging to control Monochmus beetles that transmit pine wood nematode, Bursaphelenchus xylophilus. We evaluated potential lethal or sublethal effects of thiacloprid on survival and behavior of a carpenter ant species, Camponotus japonicus Mayr. Field-collected ant colonies were directly exposed to several food items, such as thiacloprid-addicted Monochmus beetles, 10% sugar watered cotton balls contaminated by thiacloprid concentrations, and 10% sugar water. Dead beetle bodies caused no apparent adverse effect through dietary exposure in general, although a few ants were died with paralysis at colony level experiment. At individual level, most ant workers were died within 10 days compared to control group. In contrast, dietary exposure of ants to thiacloprid concentrations showed significant lethal effect with paralysis and impaired walking, especially at 10 and 50 mg/L thiacloprid concentrations. Some intoxicated ants recovered within a few days in 10 and 50 mg/L thiacloprid concentrations, but intoxicated ants were generally shown to be less responsible to enemy ants with low aggressive behavior. Implications for predicting hazards of thiacloprid to beneficial arthropods in pine forests are discussed.

The nematode Bursaphelenchus xylophilus is the causal pathogen of pine wilt disease in Korea. Currently, it is reported that B. mucronatus also has a low pathogenicity. Despite this difference in pathogenicity, it is very difficult to differentiate these two species due to similarity in morphological characters. The sequence variation of internal transcribed spacer (ITS) and intergenic spacer (IGS) regions of ribosomal DNA has been used for species identification and phylogeny of B. xylophilus and B. mucronatus. But, the IGS region sequence data of B. mucronatus has been only reported in Portugal. In this study, We analyzed genetic variation on ITS and IGS regions of B. xylophilus and B. mucronatus (Asian genotype, European genotype) based on rDNA gene sequences, and conducted phylogeography using TCS network. When the each isolates was determined the phylogenetic analysis using the neighbor-joining method, Bursaphelenchus species were divided into each groups, and showed low variation within each species. In the TCS analysis, The isolate of B. xylophilus and B. mucronatus (Asian genotype and European genotype) were divided into each groups and confirmed slightly genetic distance within species. B. mucronatus European genotype has possibility of new species different with Asian genotype.

In Korea, two sawyer beetle species belonging to genus Monochamus are vectors that transmits the pine wood nematode. In this study, we surveyed the seasonal change of spatiotemporal distribution of Monochamus alternatus and Monochamus saltuarius. A total of 360 multi-funnel traps were placed in three different regions, such as Gapyeong, Yeongdeok, and Jeju. In M. saltuarius, we collected 1,496 individuals and 709 individuals in Gapyeong and Yeongdeok, respectively, and in M. alternatus 348 individuals and 113 individuals were collected in Jeju and Yeongdeok, respectively. We analysed their patterns in spatiotemporal distribution using by global spatial autocorrelation. In M. alternatus, spatially clustering on Jeju was observed in July and August, and M. saltuarius in a site of Gapyeong was clustered. These spatiotemporal distributions of two Monochamus species may be probably related to their behavior, such as maturation feeding and dead tree searching.

Monochamus alternatus and M. saltuarius were reported as the vectors of Bursaphelenchus xylophilus, pine wood nematode in Korea. According to Kwon et al. (2006), each of 2 species has occupied their own regional distribution : M. saltuarius in southern part including Jeju island and M. alternatus in mid-northern part of Korean peninsula. We measured the supercooling point (SCP) of 2 species (laboratory-reared populations) by each of developmental stages. The SCPs of 2nd, 3rd and 5th instar larvae of M. saltuarius were -7.68±0.19℃, -7.02±0.69℃, -4.93±1.34℃ each of stages. On the other hand, the SCPs of 3rd, 4th, 5th instar larvae and pupae of M. alternatus. were -4.46±1.12℃, -5.94±1.33℃, -7.83±1.44℃, -9.53±1.78℃ each of stages. The SCPs of M. saltuarius larvae generally was lower than that of M. alternatus. The pupae of M. alternatus and 2nd instar larvae of M. saltuarius had the lowest SCP among measured samples. On the other hand, the highest SCP were recorded in 2nd and 5th instar larvae, each. This result shows that regional distribution of 2 beetles may be associated with the adaptation capacity to low temperature represented by the SCP as well as the developmental temperature. However, beetles experimented were not collected from pine forest fields. In further study, we are planning experiments with field populations and all developmental stages.

Pine wilt disease (PWD) is one of the most serious forest tree diseases and now the damaged area covers 7,877 ha and more than 60 cities in Korea. The causal agent of PWD, Bursaphelenchus xylophilus is transmitted by the insect vectors, pine sawyers (Monochamus spp.). Spread of PWD damage and control effect for PWD are directly related to dispersal potential of Monochamus spp. vectors. Recently, harmonic radar technology has been applied as a means for tracking individual dispersal of tagged insects. Tracking insects with harmonic radar system is similar to a mark-and-recapture technique, in which an organism is tagged, released, and recaptured. Our objective is to develop a harmonic radar tracking system by using lightweight tags to investigate dispersal of Monochamus spp., insect vectors. The harmonic radar system consists of two parts: a commercially available, light-weight (1.6 kg), handheld transceiver (RECCO Rescue System) and a small tag that reflects the transceiver signal (917 ㎒) at harmonic frequencies (1,834 ㎒). A tag consists of a miniature diode (Toshiba ISS350) with wires (36-gauge tinned copper) soldered using conductive epoxy on each side to produce a dipole antenna. These tags were evaluated in an open field to assess their maximum detection range, and at tag length of 13 ㎝, the tag was detected at >100 m.

Background: Rehmannia glutinosa is a perennial herb belonging to the family Scrophulariaceae. Its root has been utilized as a traditional medicine but the aerial parts (flower, flower stalk, leaf) are not used. We aimed to determine the content of three compounds [aucubin, catalpol, and γ-aminobutyric acid (GABA)] in the different organs of R. glutinosa cultivars (Dakang, Tokang, and Suwon 9) Methods and Results: The flower, flower stalk, leaf, and root of R. glutinosa were harvested at the end of August. The aucubin and catalpol contents were analyzed by LC/MS, whereas the GABA content was analyzed by GC/MS. The aucubin content was the highest in the leaf, while catalpol and GABA were the highest in the flower. The aucubin contents of leaf in Dakang, Tokang, and Suwon 9 were 1.43, 0.81, and 1.07㎎/g, respectively. The catalpol contents of flower in Dakang, Tokang, and Suwon 9 were 41.06, 28.78, and 37.48㎎/g, respectively, the GABA contents were 0.79, 0.76, and 0.65㎎/g, respectively. Conclusions: The aucubin, catalpol, and GABA contents were higher in the leaf and flower than that in the root. This study show that R. glutinosa leaf and flower can be used as a potential supplement.

Background: The roots of Atractylodes macrocephala Koidzumi contain atractylone, which is used to suppress appetite and indigestion caused by gastrointestinal disturbance. The present study was conducted to investigate the effect of several organic compost on the growth and root yield of A. macrocephala with organic fertilizer.Methods and Results:When organic fertilizer was applied basally, the average yield of 10 a was 184.6 ㎏ in the HA (Hwanggeumjidae, organic material mix), 171.3 ㎏ in the GG (Gyunbaeyangchegreen, bacterial cuture filtrate) and 175.0 ㎏ in the CF (Customary fertilization, control) each other in practice of CF had no statistical significance. Atractylenolide I was significantly greater in the HA (0.036%) than the GG (0.034%) or CF (0.023%). With regard to the amount of organic fertilzer, 10 a yield ws the most common of 203.0 ㎏ at 2.0 times of the organic 1 (HA), conventional fertilization of 134.0 ㎏ and 173.0 ㎏ of no application was a statistically significant. Organic fertilizer 1 was 1.5 to 2.0 times, organic fertilizer was 2 to 1.5 times that were most suitable.Conclusions:The results of the present study indicated that HA and GG are the most suitable for the organic cultivation of A. macrocephala. The content of atractylone I was highest under the HA treatment and lowest under the CU (Chamjoa, oil cake), TG (Totogreen, plant oil cake) and HG (Heuksalgreen, Castor oil cake) treatment.

Background: This study was conducted to investigate the effects of germination temperature, storage container and storage temperature on Scrophularia buergeriana and Scrophularia takesimensis seeds.Methods and Results: Seed lengths of both species were 0.8 ㎜, while seed width differed, with S. buergeriana measuring 0.5 ㎜ and S. takesimensis measuring 0.4 ㎜. The seeds of S. buergeriana were packaged in paper containers under room temperature (15°C), cold temperature (4°C), and freeze temperature (−20°C). These seeds exhibited around 80% germination rate at temperatures between 15°C and 30°C. The germiantion rate of S. takesimensis, on the other hand, differed significantly at different germination temperatures. Seeds of S. takesimensis which were packaged in vinyl and paper containers and stored under room and cold temperatures, exhibited around 80% germination rate at 15°C. However, the germination rate of freeze-stored seeds were decreased to lower than 20% at germination temperatures of 15°C, 25°C and 30°C germiantion conditions. The rate of germination showed a low positive to a significantly negativie correlation with the other factor that were determined to evaluate the germination performance.Conclusions: This study elucidates the most suitable germination and storage conditions to increase the germination rate for the two species of Scrophularia buergeriana and Scrophularia takesimensis needs to be stored in paper containers under cold temperature and requires a temperature of 20°C for germination. On the other hand, S. takesimensis in vinyl containers need to be stored at room temperature and those in paper containers at cold temperature, and a temperature of 15°C is required for germination.

Background : In previous studies, adlay seeds showed a prevalence of diversified fungal flora with the predominant fungal genera being Fusarium (45.6%) and In vitro test showed that fungal toxins like Fumonisin and Zearalenone were produced by Fusarium fujikuroi, F. asiaticum and F. graminearum. Because of this we performed experiments to selecting disinfective chemicals for controlling the Fusarium contamination in the adlay seed. Methods and Results : We carried out the chemical efficacy test such as seed disinfectants selection test appling before planting and pesticides selection test using in the earing season. In the present study, eleven different commercially available seed disinfectant were applied to the adlay seeds. Among 11 seed disinfectants, Hexaconazole+Prochloraz emulsifiable concentrate (EC) and Prochloraz emulsifiable concentrate (EC) had control value of 80% or above against Fusarium species tested. In the pesticides selection test, seven different commercially available pesticides for Fusarium blight (Scab) control were applied and we observed that Metconazole suspension concentrate(SC) strongly inhibited the mycelial growth of 10 Fusarium species all. Conclusion : From the above results, we selected Hexaconazole+Prochloraz EC and Prochloraz EC as a seed disinfectants and Metconazole SC as a pesticide using in the earing season for Scab control.

Background : This experiment was conducted to select GAP applying seed disinfectants in Astragalus membranaceus and Platycodon grandiflorum. Methods and Results : We carried out the chemical efficacy and injury test. For the efficacy test, we investigated fungal detection rate by seed disinfectants and for the chemical injury, we investigated germination rate and emergence rate by seed disinfectants in reference amount and fold amount. These experiments carried out two times. The results obtained are as follows. In the experiment for seed disinfectants selection of Astragalus membranaceus, all tested chemicals such as Tebuconazole emulsifiable concentrate(EC), Thiophanate-methyl + Triflumizole wettable powder(WP), Prochloraz copper chloride complex+Tebuconazole suspension concentrate(SC), Prochloraz emulsifiable concentrate(EC), Fludioxonil wetting liquid(WL) and Hexaconazol+Prochloraz emulsifiable concentrate(EC) had control value of 80% or above against seed contaminated fungi. However two chemicals such as Tebuconazole EC and Prochloraz copper chloride complex+Tebuconazole SC and two chemicals such as Prochloraz EC and Hexaconazol+Prochloraz EC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. In the case of seed disinfectants selection of Platycodon grandiflorum, Prochloraz copper chloride complex+Tebuconazole SC, Prochloraz EC and Hexaconazol+Prochloraz EC had control value of above 80% against seed contaminated fungi except Thiophanate-methyl+Triflumizole WP and Fludioxonil WL. However Hexaconazol+Prochloraz EC and Prochloraz copper chloride complex+Tebuconazole SC exhibited chemical injury significantly in reference amount and in fold amount respectively, compared to non treated control. Conclusion : From the above results, we finally selected three items of seed disinfectants including Thiophanate-methyl+Triflumizole WP and Fludioxonil WL in Astragalus membranaceus and Prochloraz emulsion in Platycodon grandiflorum.

Background : Cynanchum wilfordii and Cynanchum auriculatum belong to the Asclepiadaceae family and appear morphologically similar. In order to discriminate them, it is needed to find the presence of sap and the leaf shapes: C. auriculatum has a blade ovate leaf comparing to C. wilfordii. However, in the herbal medicine market, they have been handled as cut and dried roots. Due to their similar morphology, it is limited to distinguish the roots of C. wilfordii and C. auriculatum. Recently in Korea, it has been a critical issue to misuse these two roots in the herbal market and food industry. Thus, it is required to establish a robust tool for the discrimination and quality control of them. Methods and Results : To separation and characterization of flavor compounds, C. wilfordii and C. auriculatum samples were analyzed by head space solid phase micro extraction (HSS) fiber coupled with gas chromatography-mass spectrometry using Rtx-5MS (30 m x 0.25 mm x 0.25 μm) column. As a result, We have identified compounds of a few hundred in aliphatic aldehydes and aliphatic alcohols, alkenes and acids, aromatic compounds, aromatic compounds containing nitrogen & sulfur, etc,. In particular, The aliphatic and aromatic compounds had been clearly separated on the second dimensional direction by using two-dimensional GC. Conclusion : The volatile flavor compounds of C. wilfordii and C. auriculatum could easily analyzed without pre-treatment with improved resolution and sensitivity using HSS-GCxGC-TOFMS. We have identified compounds of a few hundred in C. wilfordii and C. auriculatum sample. And It was more accurately qualitative confirmed with separation of GCxGC and TSD. We have confirmed the PCA and PLS-DA Plot that was classified depending on C. wilfordii and C. auriculatum through multivariate statistical analysis of the identified flavor compounds.

Background : Angelica dahurica Bentham et Hooker, Angelica gigas Nakai, Ostericum koreanum Maximowicz and Peucedanum japonicum Thumberg　are a major medicinal plant in north Geungbuk province. Using medicinal plants are impotant it`s ingredient. Dry condition and stroage method are not standard manual. The ingredient variation of dry condition and stroage method were not researched. Methods and Results : Using plant material were cutivated on Gyongsangbukdo Bonghwa area. It were studied ingredient variation after dry and storage condition by HPLC methods. Major ingredient of Angelica gigas Nakai are decurusin, decurusinangelate. Heated air bulk dry get more decursin than natuarl dry and decurusinangelate of natural bulk dry was higher than heated air bulk dry. Major ingredient of Ostericum koreanum Maximowicz are imperatorin and isoimperatorin.. Imperatorin of Ostericum koreanum was highest peak on 50℃ heated-air dry after plastic bag sorage and isoimperatorin was highest peak on 40℃ heated-air dry after mountain cultivation. Imperatorin is a major ingredient Angelica dahurica Bentham et Hooker. Heated air bulk dry get more decursin and decursinangelate than natuarl dry and small heated-air dry. Peucedanol-7o_glucoside is a major ingredient Peucedanum japonicum Thumberg. Natural bulk dry get more peucedanol-7o_glucoside than heated-air bulk dry. Conclusion : Ingredient of Angelica dahurica Bentham et Hooker, Angelica gigas Nakai, Ostericum koreanum Maximowicz are different under various cutivation, drying method, storage. Diffent Ingedients of Angelica gigas Nakai, Ostericum koreanum Maximowicz were not accord it’s optical conditon.

Background : Platycodon grandiflorum has been used as food material and a traditional medicine in Korea. In order to develop an efficient in vitro micropropagation technique for a rare plant species and conservation for inbred line of plant breeding. Methods and Results : Plant regeneration via organogenesis and somatic embryogenesis was investigated in Platycodon grandiflorum. Leaf, stem, root tissues of 7-day-old seedlings were cultured on 1/2MS medium containing various concentration (0, 0.5, 1 and 2 ㎎/L) of IBA, BA and NAA. The results showed that 1/2MS medium supplemented with BA+NAA 2.0 ㎎/L yielded the highest callus formation ratio of 73.5%. When various tissues (leaf, stem, root) were tested on 1/2MS medium containing BA 2.0 ㎎/L+ NAA 2.0 ㎎/L for somatic embryogenesis, the optimum tissue for embryogenic shoot induction was stem tissue. Callus were cultured on MS medium containing various concentration (0, 0.5 and 1 ㎎/L) of BA and NAA. The best rooting rate was achieved by three different medium (B5, 1/2MS and MS) and 1/2MS medium cultured the highest rooting ratio (82%). Conclusion : This study suggested that above micropropagation techniques can be used for rapid multiplication as well as in vitro or in vivo conservation of the Platycodon species.

Background : In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results : A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions : These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

Background: In the herbal medicine market, Angelica gigas, Angelica sinensis, and Angelica acutiloba are all called "Danggui" and used confusingly. We aimed to assess the genetic diversity and relationships among 14 Angelica species collected from different global seed companies. Toward this aim we developed DNA markers to differentiate the Angelica species. Methods and Results: A total of 14 Angelica species, A. gigas, A. acutiloba, A. sinensis, A. pachycarpa, A. hendersonii, A. arguta, A. keiskei, A. atropurpurea, A. dahurica, A. genuflexa, A. tenuissima, A. archangelica, A. taiwaniana, and A. hispanica were collected. The genetic diversity of all 14 species was analyzed by using five chloroplast DNA-based simple sequence repeat (SSR) markers and employing the DNA fragment analysis method. Each primer amplified 3 - 12 bands, with an average of 6.6 bands. Based on the genetic diversity analysis, these species were classified into specific species groups. The cluster dendrogram showed that the similarity coefficients ranged from 0.77 to 1.00. Conclusions: These findings could be used for further research on cultivar development by using molecular breeding techniques and for conservation of the genetic diversity of Angelica species. The analysis of polymorphic SSRs could provide an important experimental tool for examining a range of issues in plant genetics.

Background : Curcuma longa L., in the family Zingiberaceae, is distributed in tropical and/or sub-tropical regions mainly in India and China. This species is commonly called turmeric, powder is used as medicinal herbs and/or flavor enhancer. It has been cultivated in southern region mainly Jindo. However, it might be possible to extend cultivation regions due to rise in average temperature. In order to select superior lines based on agronomic characteristics, we analyzed multivariate and estimated selection effects from C. longa germplasm. Methods and Results : The C. longa germplasm were cultivated in an experimental field located in Eumseong, NIHHS, RDA. The harvested roots were investigated in agronomic characteristics included in yield and then considered its relationship among the 9 germplasm by multivariate analysis method. Results from principal component analysis (PCoA) showed that it represented 70.00% and 80.44% accumulated explanation from four and five principal compounds (PC). PCoA was conducted from 9 agronomic characteristics and then correlation coefficient has been showed by analysis between each main component value and agronomic characteristics. Value of the first PC was 2.25, 24.96% explanation of total dispersion, plant height, number of rootlet and weight of rootlet were correlated with a somewhat higher level as 0.41, 0.43 and 0.52. Value of the fifth PC was 0.94, 10.43% explanation of total dispersion, the number of shoots was correlated with a higher level as 0.87. Selection effects with outstanding candidate lines including higher lines were estimated at 126.13% in yield. Conclusion : These data on multivariate based on agronomic characteristics will be give us invaluable breeding information by selection of superior lines.

Background : Astragalus membranaceus is one of the most widely used traditional medicinal herbs in Korea. Studies on the genomic of A. membranaceus resources have not been carried out so far. The present study was carried out to discriminate A. membranaceus based on genetic diversity using genomic simple sequence repeat (SSR) markers. Methods and Results : We collected 5 A. membranaceus lines: Asung, Poongsung, Am-Jecheon, Am-Sancheong, and Am-China. One hundred mg of fresh leaves were used for genomic DNA extraction using the DNeasy plant DNA isolation kit (Qiagen GmbH, Hilden, Germany). 450,449 contigs were searched for 147,766 SSR candidate loci in this study using the MicroSAtellite identification tool (MISA). We selected 949 A. membranaceus genomic SSR markers that were showed variation for the five collections in silico screening with CLC genomics workbench program. The genetic diversity of all A. membranaceus resources was analyzed using 17 SSR markers employing the DNA fragment analysis method. Based on the genetic diversity analysis, these lines were classified into four distinct groups. Conclusion : These findings could be used for further research on cultivar development using molecular breeding techniques and for conservation of the genetic diversity of A. membranaceus. Furthermore, the markers could be used for marker-assisted selection for crop breeding.