Lilium Lancifolium Thunb Research Articles

Characterization of LiMAPK gene in response to salinity stress in Tiger lily (Lilium lancifolium Thunb.)

In this study, we measured the physiological response such as relative growth rate (RGR), relative water content (RWC), chlorophyll index, net photosynthetic rate and relative electrical conductivity of Tiger lily leaves periodically, under different concentrations of salt. We isolated LiMAPK mRNA (including the 5′ UTR and 3′ UTR) (GenBank accession number JQ437268) under treatment at salt threshold concentrations by rapid amplification of cDNA ends polymerase chain reaction (RACE–PCR). The relative expression levels of LiMAPK under salt stress were analyzed by real-time fluorescence quantitative PCR. The results showed that the salt threshold concentration of Tiger Lily was 1.09 mg/mL, and the LiMAPK gene covered 1291 bp, coded 306 amino acids and the conservative domain was 627 bp long. There was a highly conserved domain (TRWYRAPE) present in the amino acid sequence of MAPK. Comparative analysis using NCBI BLASTp showed that this conservative domain was a catalytic domain in the MAPK super family. The LiMAPK expression levels in the leaves were significantly increased within 2 h after initiation of stress treatment with salt threshold concentrations and continued to rise in the next 2 h, reaching a maximum at 4 h and decreasing in the next 4 h. After 12 and 24 h of salt stress, the LiMAPK expression levels were basically stable, but still twice as high as those in the control. The results indicated that the LiMAPK gene was up-regulated by salt stress, which might be closely related to the salt-tolerance mechanism in Tiger lily.

Characterization of Conserved and Novel microRNAs in Lilium lancifolium Thunb. by High-Throughput Sequencing

MicroRNAs (miRNAs) are among the class of noncoding small RNA molecules and play a crucial role in post-transcriptional regulation in plants. Although Lilium is one of the most popular ornamental flowers worldwide, however, there is no report on miRNAs identification. In the present study, therefore, miRNAs and their targets were identified from flower, leaf, bulblet and bulb of Lilium lancifolium Thunb. by high-throughput sequencing and bioinformatics analysis. In this study, a total of 38 conserved miRNAs belonging to 17 miRNA families and 44 novel miRNAs were identified. In total, 366 target genes for conserved miRNAs and 415 target genes for novel miRNAs were predicted. The majority of the target genes for conserved miRNAs were transcriptional factors and novel miRNAs targeted mainly protein coding genes. A total of 53 cleavage sites belonging to 6 conserved miRNAs families and 14 novel miRNAs were identified using degradome sequencing. Twenty-three miRNAs were randomly selected, then, their credibility was confirmed using northern blot or stem-loop qRT-PCR. The results from qRT-PCR analysis showed the expression pattern of 4 LL-miRNAs was opposite to their targets. Therefore, our finding provides an important basis to understand the biological functions of miRNAs in Lilium.

Ribosomal DNA locus variation and REMAP analysis of the diploid and triploid complexes ofLilium lancifolium

Lilium lancifolium Thunb. (2n = 2x = 24) is a cytologically conspicuous species with both diploids and triploids in nature. Cytological and molecular genetic analyses were carried out in both diploids and triploids that were collected from 55 geographical locations in Korea, Japan, and China. While the 5S rRNA gene loci were located at duplicated loci on the long arm of chromosome 2, the 45S rRNA gene loci were present in chromosomes 1, 2, 4, 6, 7, and 11. While the loci on chromosomes 1 and 7 were constant, the loci on chromosomes 2, 4, 6, 7, and 11 were variable in some plants so that the L. lancifolium accessions were grouped into 7 cytotypes in diploids and 12 cytotypes in triploids. REMAP marker analysis revealed that the diploids were classified into seven clusters, and the triploids were classified into a large cluster. Geographic, cytological, and genetic differentiations were not related in both the diploid and triploid accessions of L. lancifolium. Thus, current genetic variations occurred prior to the geographic differentiation in both diploids and triploids, and the 45S rDNA cytotype variations occurred after geographic differentiation in the current habitats of L. lancifolium.

A novel biosurfactant produced by Aureobasidium pullulans L3-GPY from a tiger lily wild flower, Lilium lancifolium Thunb.

Yeast biosurfactants are important biotechnological products in the food industry, and they have medical and cosmeceutical applications owing to their specific modes of action, low toxicity, and applicability. Thus, we have isolated and examined biosurfactant-producing yeast for various industrial and medical applications. A rapid and simple method was developed to screen biosurfactant-producing yeasts for high production of eco-friendly biosurfactants. Using this method, several potential niches of biosurfactant-producing yeasts, such as wild flowers, were investigated. We successfully selected a yeast strain, L3-GPY, with potent surfactant activity from a tiger lily, Lilium lancifolium Thunb. Here, we report the first identification of strain L3-GPY as the black yeast Aureobasidium pullulans. In addition, we isolated a new low-surface-tension chemical, designated glycerol-liamocin, from the culture supernatant of strain L3-GPY through consecutive chromatography steps, involving an ODS column, solvent partition, silica gel, Sephadex LH-20, and an ODS Sep-Pak cartridge column. The chemical structure of glycerol-liamocin, determined by mass spectrometry and nuclear magnetic resonance spectroscopy, indicates that it is a novel compound with the molecular formula C33H62O12. Furthermore, glycerol-liamocin exhibited potent biosurfactant activity (31 mN/m). These results suggest that glycerol-liamocin is a potential novel biosurfactantfor use in various industrial applications.

Occurrence and Survival of Autotriploids in Natural Diploid Populations of Lilium lancifolium Thunb.

Occurrence and Survival of Autotriploids in Natural Diploid Populations of Lilium lancifolium Thunb.

Structural characterisation, physicochemical properties and antioxidant activity of polysaccharide from Lilium lancifolium Thunb.

A novel polysaccharide fraction (LP2-1) was isolated and purified from the edible bulbs of Lilium lancifolium Thunb. by DEAE cellulose chromatography and Sephacryl S-400 size-exclusion chromatography. The structural characterisation, physicochemical properties and antioxidant activity of LP2-1 were investigated. The results showed that LP2-1 had an average molecular weight of approximately 8.52×103kDa and was mainly composed of l-rhamnopyranose, d-arabinofuranose, d-glucopyranose and d-galactopyranose in the molar ratio of 1.88:2.13:1.00:2.50, and major functional groups of LP2-1 were COO and OH. The viscoelastic properties of LP2-1 systems exhibited a gel-like behaviour, with storage modulus higher than loss modulus, and both moduli increased with increasing Ca2+ concentration. In addition, LP2-1 had DPPH and hydroxyl radicals scavenging activities, and also had the strong reducing power and chelating activity on ferrous ion. These results suggest that LP2-1 has good antioxidant activity and can be used in food industry.

Lilium lancifolium Thunb. extract attenuates pulmonary inflammation and air space enlargement in a cigarette smoke-exposed mouse model

Ethnopharmacological relevanceLilium lancifolium Thunb. (Liliaceae) has long been used as a traditional medicine in Korea and China to treat bronchitis, pneumonia, and other pulmonary ailments. Aim of the studyCigarette smoke (CS) is a major risk factor for the development of pulmonary inflammatory response; it also triggers pulmonary alveoli enlargement. In the present study, we investigate the effects of Lilium lancifolium Thunb. root extract on pulmonary inflammatory responses in a CS-exposed mouse model. Materials and methodsWater extract of Lilium lancifolium Thunb. root was fed to C57BL/6 mice prior CS exposure every day for 3 weeks. The numbers of macrophages and neutrophils in bronchoalveolar lavage fluid (BALF) were counted. The relative inflammatory factors, tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), interleukin-1 beta (IL-1β), monocyte chemotactic protein-1 (MCP-1), and matrix metalloproteinase-12 (MMP-12) were measured by real-time PCR, ELISA, or Western blot analysis. The average alveoli size was determined by lung histology. ResultsLilium lancifolium Thunb. root extract was found to significantly inhibit the numbers of macrophages and neutrophils in BALF due to CS exposure. Lilium lancifolium Thunb. root extract also reduced the protein secretion levels of TNF-α, IL-6, IL-1β, and MCP-1 in BALF and the RNA expression levels of TNF-α, IL-6, IL-1β, MCP-1, and MMP-12 in lung tissue compared with mice only exposed to CS. Moreover, MMP-12 in serum was down regulated in Lilium lancifolium Thunb. root extract treated mice compared with CS-exposed mice. Finally, a morphometric analysis of the lungs of Lilium lancifolium Thunb. root extract treated mice demonstrated a significant reduction in airspace size compared to mice only exposed to CS. ConclusionOur results show that Lilium lancifolium Thunb. root extract reduces lung inflammation and airspace enlargement in a CS-exposed mouse model. These data indicate that Lilium lancifolium Thunb. root extract is a therapeutic candidate for pulmonary inflammation and emphysema caused by CS.

Cryopreservation of in vitro-grown apical meristems of Lilium by droplet-vitrification

In this study, apical meristems from adventitious buds of three lily ( Lilium L.) cultivars were successfully cryopreserved by droplet-vitrification. The most effective techniques were as follows. Excised meristems from in vitro plantlets which had been sub-cultured for about 2 months were cold-hardened at 4 °C for 1 week, precultured on MS medium supplemented with 0.3 mol L −1 sucrose for 2 days, osmoprotected in loading solution for 20–40 min at room temperature and then soaked in PVS2 solution for 90–120 min at 0 °C, frozen in microdroplets of vitrification solution placed on aluminium foils, which were immersed rapidly in liquid nitrogen. The meristems were then rapidly rewarmed by dilution solution, transferred to regeneration medium and stored in the dark for two weeks at 20 °C, and then cultured under white fluorescent light at an intensity of 2000 lux, with a 16 h photoperiod at 20 °C. The highest post-thaw survival percentages of three cultivars ‘Siberia’ (Lilium × siberia), Lilium lancifolium Thunb. and ‘Snow Queen’ Lilium × longiflorum were 65.0%, 83.8% and 43.3%, and regeneration percentages were 62.0%, 67.6% and 35.0%, respectively. The study demonstrated that cryopreservation by droplet-vitrification increased survival and regeneration percentages of certain lily cultivars compared with vitrification. Thus to cryopreserve lily meristems, droplet-vitrification method is preferable to the vitrification method.

Lilium lancifolium is Discovered as a New Host of Botryosphaeria parva in Georgia

A new disease on tiger lily (Lilium lancifolium Thunb.) was discovered in a residential settling in Georgia. Koch's postulate and molecular techniques confirmed that Botryosphaeria parva was the causal agent. This is the first report of B. parva on tiger lily. Accepted for publication 8 September 2006. Published 27 November 2006.

Particle Bombardment에 의해 전처리 된 참나리(Lilium lancifolium Thunb.) 캘러스의 Agrobacterium tumefaciens을 통한 형질전환

형질전환 효율을 높이기 위하여, NPTII와 GUS유전자를 포함하고 있는 pIG121Hm을 미세 입자에 코팅하여 particle bombardment를 수행한 후 pIG121Hm가 도입된 Agrobacterium tumefaciens EHA101와 3일간 공동 배양하였다. 그 후 캘러스를 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin, 그리고 200mg/L carbenicillin이 포함된 MS배지로 옮겨 4주간 성장시킨 후 kanamycin저항성 캘러스를 선발하여 GUS 발현을 관찰하였고, PCR 분석을 통하여 700 bp의 NPT II 유전자가 도입된 것을 확인하였다. Particle bombardment 후 Agrobacterium과 공동배양 한 처리구가 Agrobacterium과 공동배양만 한 처리구보다 GUS발현 분석에 의한 형질전환 효율이 3배정도 더 높았다 형질전환 된 재분화 식물체를 얻기위해 다시 선발된 kanamycin저항성 캘러스는 1mg/L NAA와 1mg/L BAP가 포함된 재분화 배지에 옮겨져 4주 후 형질전환 된 소인경을 형성하였다. To improve transformation efficiency, the callus of Lilium lancifolium Thunb. were bombarded by particles coated with pIG 121 Hm which include NPT II and GUS genes, and then cocultivated with Agrobacterium tumefaciens EHA101 which contain pIG121Hm binary vector, carrying neomycin phosphotransferase (NPT II) and <TEX>$\beta$</TEX>-Glucuronidase (GUS) genes. Three days after cocultivation with Agrobacterium tumefaciens and particle bombardment, the callus clusters were transferred to MS medium containing 1mg/L 2,4-D, 0.1mg/L BAP, 100mg/L kanamycin and 200mg/L carbenicillin. Four weeks after transfer to the selection medium, GUS expression was detected and PCR analysis revealed the presence of NPT II fragment of the expected size (700 bp) in the transformed callus. The GUS expression from Agrobacterium-mediated transformants after particle bombardment increased to over 3-folds compared with that of callus cocultivated with Agrobacterium tumefaciens without particle bombardment. The callus clusters containing kanamycin resistant gene were transferred to MS medium containing 1mg/L NAA and 1mg/L BAP. Somatic embryos were developed in four weeks and microbulbs expressing GUS were formed.

Cytogenetic Behavior, Chromosomal Differentiations, and Geographic Distribution in Lilium lancifolium (Liliaceae)

Abstract Lilium lancifolium Thunb, is the only polyploid species complex consisting of 2x (2n=24) and 3x (2n=36) forms in the genus, both of which are highly self‐sterile. They are cytologically differentiated into various nucleolar types by number and combination of four kinds of nucleolar chromosomes. The 3x form is widespread from the North‐East district of China to the Japanese Islands through the Korean Peninsula. However, the 2x form is localized and grows singly or together with the 3x form in the narrow zone from southern Korea to the Tsushima Islands, Japan. The 2x form reproduces by sexual as well as by asexual means, but a sterile 3x form has asexually persisted to the present. Generally speaking, the 3x form is the allotriploid which has originated from hybridization between a 2x form and a closely related species such as L. maximowiczii in the remote past. In the present study, another possibility for the presence of the autotriploid form is newly proposed based on its karyotype and geographic distribution.