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In order to explore the response mechanism of Lilium pumilum (L. pumilum) to saline–alkali stress, we successfully cloned LpGDSL (GDSL lipase, Gly-Asp-Ser-Leu) from L. pumilum. The qRT-PCR results indicated that the LpGDSL expression was higher in the leaves of L. pumilum, and the expression of the LpGDSL reached the highest level at 12 h in leaves under 11 mM H2O2, 200 mM NaCl, 25 mM Na2CO3, and 20 mM NaHCO3. The bacteriophage overexpressing LpGDSL was more tolerant than the control under different NaHCO3 contents. Overexpressed and wild-type plants were analyzed for phenotype, chlorophyll content, O2− content, H2O2 content, lignin content, and so on. Overexpressed plants had significantly higher resistance than the wild type and were less susceptible to saline–alkali stress. The yeast two-hybrid and BiFC assays demonstrated the existence of an interaction between LpGDSL and LpBCP. The yeast one-hybrid assay and transcriptional activation assay confirmed that B3 transcription factors could act on LpGDSL promoters. Under saline–alkali stress, L. pumilum will promote the expression of LpGDSL, which will then promotes the accumulation of lignin and the scavenging of reactive oxygen species (ROS) to reduce its damage, thus improving the saline–alkali resistance of the plant. [ABSTRACT FROM AUTHOR]

Lilium pumilum DC (L. pumilum DC) plays an important role in the rational utilization of salinized soil. To explore the molecular mechanism of salt-tolerant L. pumilum, the LpMYB4 was cloned. LpMYB4 close relationship with Bambusa emeiensis and Zea mays MYB4 throughout the phylogenetic tree construction. LpMYB4 protein was found to be localized in the nucleus. Prokaryotic and eukaryotic bacterial solution resistance experiments proved that the exogenous introduction of LpMYB4 made the overexpression strains obtain better survival ability under saline-alkaline stress. Compared with wild-type plants, tobacco plants overexpressing LpMYB4 had better growth and lower leaf wilting and lodging, the content of chlorophyll was higher, the content of hydrogen peroxide and superoxide anion was lower, the activity of peroxidase and superoxide dismutase was higher and the relative conductivity was lower under saline-alkaline stress. The analysis of seed germination and seedling resistance of transgenic plants under salt stress showed that LpMYB4 transgenic seeds were more tolerant to salt stress during germination and growth. Yeast two-hybrid and two-luciferase complementation experiments showed that LpMYB4 interacted with yeast two-hybrid and LpGPX6. The analysis of the role of LpMYB4 in improving plant saline-alkali resistance is helpful to the transformation of plant germplasm resources and has great significance for agriculture and sustainable development.

Lilium pumilum plays an important role in resistance breeding of lily because of its versatile value and strong stress resistance. It is of great significance to study the conservation of its resources. To improve its vitality after cryopreservation, the effect of abscisic acid (ABA) on the viability of cryopreserved shoot tips and its mechanism were studied based on the establishment of cryopreservation-vitrification protocol of Lilium pumilum. When 25 μM abscisic acid was added into the preculture medium, the survival rate after cryopreservation was increased by 22.73% and the regeneration rate by 11.48%, reaching 93.64% and 63.33%, respectively. Adding exogenous abscisic acid increased the contents of endogenous osmotic regulatory substances proline, soluble sugar and soluble protein and reduced the activities of caspase-3-like and caspase-9-like enzyme that characterize programmed cell death (PCD). This suggests that the addition of abscisic acid during preculture phase of cryopreservation improved viability of shoot tips by affecting osmotic stress and programmed cell death. Key message: A simple and efficient cryopreservation procedure for the shoot tips of Lilium pumilum was established. Exogenous abscisic acid increased the viability of shoot tips by regulating osmotic stress and programmed cell death. [ABSTRACT FROM AUTHOR]

Copyright of Bulletin of Botanical Research is the property of Bulletin of Botanical Research Editorial Department and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

(1) Background: The growth of plants is impacted by salinity and alkali, Lilium pumilum (L. pumilum) is an ornamental plant with strong resistance to salinity and alkali, while the LpPsbP gene is helpful to fully understand the Saline-Alkali tolerance of L. pumilum. (2) Methods: Gene cloning, bioinformatics analysis, expression of fusion protein, determination of physiological indices of plant after Saline-Alkali stress, yeast two-hybrid screening, luciferase complementation assay, chromosome walking to obtain the promoter sequence, and then analyzed by PlantCARE. (3) Results: The LpPsbP gene was cloned and the fusion protein was purified. The transgenic plants had higher Saline-Alkali resistance than the wild type. A total of eighteen proteins interacting with LpPsbP were screened, and nine sites in the promoter sequence were analyzed. (4) Conclusion: Under Saline-Alkali or oxidative stress, L. pumilum will promote the expression of LpPsbP, which will then directly scavenge reactive oxygen species (ROS) in order to protect its photosystem II, reduce its damage, and thus improve the Saline-Alkali resistance of the plant. Moreover, according to some of the literature and the following experiments, two additional speculations are developed on the mechanisms of how two newly found objects, namely jasmonic acid (JA) and FoxO protein, could be involved in ROS scavenging processes were made. [ABSTRACT FROM AUTHOR]

[Display omitted] • The LpNAC5 isolated from Lilium pumilum was overexpressed in tobacco plants. • Transgenic tobacco overexpressing LpNAC5 showed significant tolerance to drought, salt, and alkali stresses. • The expression of LpNAC5 was affected by abscisic acid and might be involved in ABA-signaling pathway. NACs (NAM、ATAF1/2、CUC1/2), as a large family of plant transcription factors, are widely involved in abiotic stress responses. This study aimed to isolate and clone a novel stress-responsive transcription factor LpNAC5 from Lilium pumilum bulbs. Drought, salt, alkali, and ABA stresses induced the expression of LpNAC5. Transgenic tobacco plants overexpressing LpNAC5 were generated using the Agrobacterium- mediated method to understand the role of this factor in stress response. These plants exhibited increased tolerance to drought, salt, and alkali stresses. The tobacco plants overexpressing LpNAC5 showed strong drought, salt, and alkaline tolerance. Under the three abiotic stresses, the activities of antioxidant enzymes were enhanced, the contents of proline and chlorophyll increased, and the contents of malondialdehyde decreased. The functional analysis revealed that LpNAC5 enabled plants to positively regulate drought and salt stresses. These findings not only provided valuable insights into stress tolerance mechanisms in L. pumilum but also offered a potential genetic resource for breedi. [ABSTRACT FROM AUTHOR]

Lilium Pumilum with wide distribution is highly tolerant to salinity. The blue copper protein LpCPC (Lilium pumilum Cucumber Peeling Cupredoxin) gene was cloned from Lilium pumilum, which has the conserved regions of type I copper protein. Moreover, LpCPC has the closest relation to CPC from Actinidia chinensis using DNAMAN software and MEGA7 software. qRT-PCR indicated that LpCPC expression was higher in root and bulb of Lilium pumilum, and the expression of the LpCPC gene increased and reached the highest level at 12 h in bulbs under 20 mM NaHCO3. The transgenic yeast was more tolerant compared with the control under NaHCO3 stress. Compared with the wild type, overexpressing plants indicated a relatively lower degree of wilting. In addition, the chlorophyll content, soluble phenol content, and lignin content of overexpressing lines were higher than that of wild-type, whereas the relative conductivity of overexpressing plants was significantly lower than that of wild-type plants. Expression of essential genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. Transgenic lines had much higher levels of CCR1 and CAD, which are involved in lignin production, compared with wild-type lines. The yeast two-hybrid technique was applied to screen probable interacting proteins interacting with LpCPC. Eight proteins interacted with LpCPC were screened, and five of which were demonstrated to be associated with plant salinity resistance. Overall, the role of gene LpCPC is mediating molecule responses in increasing saline-alkali stress resistance, indicating that it is an essential gene to enhance salt tolerance in Lilium pumilum.

Lilium pumilum is an important ornamental, culinary and medicinal bulbous plants with salt tolerance. However, salt tolerance of lily, particularly the bulb, has been studied relatively little, which brings challenges to the cultivation of lily varieties with high salt tolerance. Here, we performed transcriptome sequencing on the bulb organs of L. pumilum under salt stress treatment, analyzed differential gene expressed levels and then identified several key genes associated with salt stress tolerance at genome-wide scale. For the first time, we revealed the obvious response against salt stress for L. pumilum bulb organs, while distinct from those for root organs. Several key genes obtained through transcriptome analysis and DEG screening include NF-YB3 transcription factor, metallothionein type 2 protein, vicilin like seed storage protein and bidirectional sugar transporter SWEET14. Rather than typical ROS scavengers like superoxide dismutase, peroxidase, and glutathione transferase, non-typical ROS scavengers such as the metallothionein type 2 protein, and vicilin like seed storage protein were upregulated in our work. The bidirectional sugar transporter SWEET14 protein and the hormone signaling proteins such as E3-ubiquitin protein ligases, PYL4 and protein phosphatase 2C were also upregulated, suggesting the role of sugars and hormones in the bulb organ responses to salt stress. Co-expression analysis of the DEGs further confirmed that NF-YB3 transcription factor acted as a hub gene, suggesting that salt stress can promote flowering of L. pumilum. Taken together, we identified important candidate genes associated with salt tolerance of the L. pumilum bulb organs, which may provide the excellent basis for further in-depth salt tolerance mechanisms of the lily bulbs. [ABSTRACT FROM AUTHOR]

Lilium pumilum is an excellent wildflower germplasm resource with high resistance to salinity stress. The gene LpBCP plays an important role in salinity tolerance of L. pumilum. Studying the molecular mechanism of salinity resistance in L. pumilum will provide insights into multiple aspects, including breeding better varieties, environmental protection, improving soil conditions, etc.Conventional methods were used to determine different physiological indicators of Nicotiana benthamiana after NaHCO3 treatment, i.e. chlorophyll content, soluble phenol content and lignin content. RT‐qPCR was carried out to find expression of LpBCP in different organs and under abiotic stresses. DAB was used to detect H2O2 in leaves in situ. A yeast two‐hybrid system was used to screen for LpBCP interacting proteins.LpBCP was cloned from bulbs of L. pumilum. The highest expression of LpBCP was in roots and bulbs of transgenic plants. LpBCP‐overexpressed plants showed less wilting, compared to WT plants. LpBCP transgenic plants have higher chlorophyll, soluble phenol and lignin content, and lower relative conductivity under 500 mM NaHCO3 stress. In addition, H2O2 scavenging in transgenic plants was much improved, indicating increased resistance to NaHCO3 stress. Thirteen LpBCP‐interacting proteins were screened using the yeast two‐hybrid method and five were associated with salt stress.Based on our findings, LPBCP could be a key gene that can be used to improve L. pumilum salt tolerance. [ABSTRACT FROM AUTHOR]

Lilium pumilum is a perennial herb with ornamental edible and medicinal value. It is an excellent wild germplasm resource with wide distribution and strong resistance. The NAC family of transcription factors is unique to higher plants. The NAC family plays a regulatory role in plant growth and development and participates in plant responses to biotic and abiotic stresses. The LpNAC17 gene of L. pumilum was cloned and transformed into tobacco to investigate the response of transgenic tobacco to salt stress. The results showed that the net photosynthetic rate and contents of chlorophyll in LpNAC17 over-expressed tobacco were higher than those in the control plants, while the stomatal conductance, transpiration rate and intercellular CO2 concentration were lower than those in the controls. The activity of superoxide dismutase, peroxidase, catalase, and the content of proline in LpNAC17 over-expressed tobacco were higher than those in the controls, while the content of malondialdehyde, superoxide anion, and hydrogen peroxide were lower than that in the control. Nitro-blue tetrazolium staining and 3,3′-diaminobenzidine tissue localization showed that the contents of O2- and H2O2 in transgenic tobacco was lower than in the controls. The expression levels of NtSOD, NtPOD, NtCAT, NtHAK1, NtPMA4, and NtSOS1 in the transgenic tobacco were higher than those in the controls. Therefore, this study provides a gene source for molecular breeding of salt-tolerant plants through genetic engineering, and lays a foundation for further research on salt-tolerant Lily. [ABSTRACT FROM AUTHOR]

Lilium pumilum is widely distributed in northeast Asia. It exhibits strong resistance and possesses high ornamental value. However, it currently lacks an efficient and stable transformation system. Therefore, we aimed to establish an effective genetic transformation system using the Agrobacterium-mediated method for L. pumilum, enabling gene transfer into the plant for gene function research and genetic engineering breeding. Our genetic transformation system achieved a transformation efficiency of 7.25 % under specific conditions: a kanamycin (Kana) concentration of 120 mg/L, 3 days of pre-cultivation, an A. tumefaciens concentration of 0.7 OD 600 , an acetosyringone (AS) concentration of 20 mg/L, and a 15-minute infection period. We investigated the function of the LpNAC6 from L. pumilum by observing phenotypic and physiological changes under stresses induced by salt, alkali, and drought. Furthermore, overexpression of LpNAC6 resulted in enhanced stress tolerance as evidenced by increased levels of SOD, POD, CAT enzymes, improved photosynthetic indices, and elevated chlorophyll contents; as well as reduced levels of MDA and reactive oxygen species (ROS). These findings demonstrate that we have successfully established a transgenic transformation method for L. pumilum while also providing essential information for cultivating stress-tolerant Lilium species and advancing our understanding of the functions of LpNAC6 in plants., Competing Interests: Declaration of Competing Interest The authors declare that they have no competing interests., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Lilium pumilum DC ( L. pumilum DC) plays an important role in the rational utilization of salinized soil. To explore the molecular mechanism of salt-tolerant L. pumilum , the LpMYB4 was cloned. LpMYB4 close relationship with Bambusa emeiensis and Zea mays MYB4 throughout the phylogenetic tree construction. LpMYB4 protein was found to be localized in the nucleus. Prokaryotic and eukaryotic bacterial solution resistance experiments proved that the exogenous introduction of LpMYB4 made the overexpression strains obtain better survival ability under saline-alkaline stress. Compared with wild-type plants, tobacco plants overexpressing LpMYB4 had better growth and lower leaf wilting and lodging, the content of chlorophyll was higher, the content of hydrogen peroxide and superoxide anion was lower, the activity of peroxidase and superoxide dismutase was higher and the relative conductivity was lower under saline-alkaline stress. The analysis of seed germination and seedling resistance of transgenic plants under salt stress showed that LpMYB4 transgenic seeds were more tolerant to salt stress during germination and growth. Yeast two-hybrid and two-luciferase complementation experiments showed that LpMYB4 interacted with yeast two-hybrid and LpGPX6. The analysis of the role of LpMYB4 in improving plant saline-alkali resistance is helpful to the transformation of plant germplasm resources and has great significance for agriculture and sustainable development.

• For improving the abiotic stress tolerance of Lilium species, identified 23 MYB genes in Lilium pumilum (salt-tolerant Lilium species) have the conserved domain such as SANT domain in common. • Phylogenetic relationships of the LpMYB transcription factors showed their diverse roles related to various developmental processes and environmental interactions in Lilium pumilium. • LpMYBs were upregulated to varying degrees under different stresses, in particular, LpMYB1 showed the significant induction to response upon salt in roots of L. pumilum. • Transgenic plants overexpressing LpMYB1 showed increased salt tolerance with higher levels of chlorophyll, proline, SOD, POD and CAT enzymes, while lower levels of MDA. The MYB family proteins play crucial roles in regulating the metabolism for plant growth and development as well as stress responses, thus, have great potential in breeding novel plant varieties with improved traits such as abiotic stress tolerance. Although the MYB genes have extensively studied in various plant species, they have not yet largely explored in Lilium species, which has high ornamental and medicinal values, limiting the practices for breeding abiotic stress tolerant Lilium species. For improving the abiotic stress tolerance of Lilium species, we identified 23 MYB genes in Lilium pumilum (salt-tolerant Lilium species) and performed comprehensive characterization of structure prediction, phylogenetic relationships, conserved motifs, and time- and tissue-specific expression patterns. Phylogenetic relationships of the LpMYB transcription factors showed their diverse roles related to various developmental processes and environmental interactions in L. pumilum. Structure prediction and conserved motif analysis revealed that LpMYB transcription factors have the conserved domain such as SANT domain in common. The LpMYB genes revealed explicit time- and tissue-specific induction pattern upon ABA, cold, PEG, and salt treatment. Particularly, LpMYB1 showed the strongest induction response upon salt in roots of L. pumilum. LpMYB1 overexpression tobacco lines further exhibited the improved salt tolerance phenotypes, such as higher levels of chlorophyll, proline, SOD, POD, and CAT enzymes while lower levels of MDA. Taken together, this study identified novel LpMYB gene family in L. pumilum and contributed to building the preliminary knowledge for further in-depth characterization of the LpMYB transcription factors as well as breeding stress-tolerant Lilium species. [ABSTRACT FROM AUTHOR]

MicroRNAs (miRNAs) play important roles in post-transcriptional regulation as well as in various biological and metabolic processes. Lilium pumilum originated from northern China with a high ornamental value, and is important for Lilium resistance breeding. The dormancy of the bulbs has a serious effect on flowering quality. In this study, the miRNAs from samples at 0 days (S1 stage), 30 days (S2 stage), 60 days (S3 stage) and 90 days (S4 stage) of cold storage were sequenced to reveal the dormancy mechanism of Lilium pumilum. Through ultrastructural observation and physiological index measurement, we found that Lilium pumilum bulbs were released from dormancy during low-temperature storage. We further conducted miRNAs sequencing; a total of 17 conserved miRNAs were identified from 12 families, two nonconserved miRNAs were found that belonged to two families, and 38 novel miRNAs were predicted. MiRNA-target pairs analysis showed that some biological processes, including antioxidant reactions, epigenetic modification and abiotic stress, were induced in response to low-temperature conditions. Some transcription factors, plant hormone signalling pathways, and enzymes involved in plant growth and development were also activated to break the bulb dormancy. The study initially revealed the complex regulatory mechanisms of miRNAs for dormancy release during low-temperature storage. Comprehensive sequence information and target gene data were obtained by miRNAs sequencing, which determined that miRNAs regulation of bulb dormancy release in low temperature. [ABSTRACT FROM AUTHOR]

A novel gram-negative, aerobic, non-spore-forming, rod-shaped and non-nitrogen fixing bacterium named strain 24NRT was isolated from wild Lilium pumilum bulbs in Fuping, Baoding City, Hebei province, PR China. The 16S rRNA gene sequences of strains 24NRT showed the highest similarity to Neorhizobium alkalisoli DSM 21826T (98.5%) and N. galegae HAMBI 540T (98.1%). Phylogenetic analysis based on 16S rRNA genes and multilocus sequence analysis (MLSA) based on the partial sequences of atpD-glnII-glnA-recA-ropD-thrC housekeeping genes both indicated that strain 24NRT is a member of the genus Neorhizobium. The average nucleotide identity between the genome sequence of strain 24NRT and that of the isolate N. alkalisoli DSM 21826T was 83.1%, and the digital DNA–DNA hybridization was 20.1%. The G+C content of strain 24NRT was 60.3 mol %. The major cellular fatty acids were summed feature 8 (C18:1ω7c and/or C18:1ω6c) and C19:0 cyclo ω8c. Based on phenotypic, phylogenetic, and genotypic data, strain 24NRT is considered to represent a novel species of the genus Neorhizobium, for which the name Neorhizobium lilium sp. nov. is proposed. The type strain is 24NRT (= ACCC 61588T = JCM 33731T). [ABSTRACT FROM AUTHOR]

NACs are one of the largest transcription factor families in plants and are involved in the response to abiotic stress. A new stress-responsive NAC transcription factor gene, LpNAC13 , was isolated from Lilium pumilum bulbs. The expression of LpNAC13 was induced by drought, salt, cold and ABA treatments. LpNAC13 overexpressing plants were generated to explore the function of LpNAC13 in response to drought and salt stress. Overexpression of LpNAC13 in tobacco displayed a reduced drought tolerance but exhibited an enhanced salt tolerance. The LpNAC13 overexpression plants had decreased antioxidant enzyme activities, content of proline and chlorophyll, increased MDA content under drought condition, the results in the LpNAC13 plants under salt condition were opposite to those under drought condition. The seed germination and root length assays of overexpression of LpNAC13 showed decreased sensitivity to ABA. Functional analyses demonstrate that LpNAC13 plays opposite roles in drought and salt stress tolerance, acting as a negative regulator of drought response but as a positive regulator of salt response in tobacco. • LpNAC13 is a stress-responsive NAC transcription factor gene, which was isolated from Lilium pumilum bulbs. • The expression of LpNAC13 was induced by drought, salt, cold and ABA treatments. • Overexpression of LpNAC13 in tobacco displayed a reduced drought tolerance but exhibited an enhanced salt tolerance. • The study of LpNAC13 provides some new view of the molecular mechanisms of drought and salt resistance in the lily family. [ABSTRACT FROM AUTHOR]

Copyright of Acta Prataculturae Sinica is the property of Acta Prataculturae Sinica Editorial Office and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)

Ferric reduction oxidase (FRO) plays important roles in biotic and abiotic stress. However, the function of ferric reduction oxidase from Lilium pumilum in response to NaHCO 3 is unknown. Here we report the functional characterization of ferric reduction oxidase 7 in Lilium pumilum (LpFRO7) in stresses. Under NaHCO 3 stress, the LpFRO7 overexpression lines exhibited lower accumulation of reactive oxygen species (ROS), higher activities in antioxidant enzyme (CAT, SOD and POD) and ferrite reductase, resulting in improved tolerance compared to the wild type (WT). In order to determine the functional network of LpFRO7 , it was confirmed by EMSA assays, Yeast one-hybrid assays and Dual luciferase reporter assays that LpbHLH115 transcription factor can bind to the promoter of LpFRO7. Yeast two-hybrid assays, BiFC, and LCI assays were performed to prove that LpFRO7 can interact with LpTrx. Combining these findings, we concluded that LpFRO7 affects plant saline-alkaline tolerance by regulating ROS homeostasis. • LpFRO7 increases the saline-alkaline tolerance in Lilium pumilum. • LpbHLH115 transcription factor could bind to the promoter of LpFRO7. • Interaction between LpFRO7 and LpTrx coordinates the homeostasis of ROS in Lilium pumilum. [ABSTRACT FROM AUTHOR]

Lilium pumilum DC. is a useful plant species not only for its showy flowers but also for its edible and medicinal values. Here we report on the complete chloroplast genome sequence of L. pumilum. The chloroplast genome is 152,573 bp in size and includes two inverted repeat regions of 52,984 bp, which is separated by a large single-copy region of 82,009 bp and a small single copy region of 17,580 bp. A total of 130 genes were predicted, including 38 tRNA, 8 rRNA, and 84 protein-coding genes. Phylogenetic analysis placed L. pumilum under the family Liliaceae.

Lily basal rot, caused by Fusarium oxysporum f. sp. lilii, is one of the most serious diseases of lily. Although the lily germplasm which is resistant to F. oxysporum has been used in disease-resistant breeding, few studies on its molecular mechanism of disease resistance have been reported. To comprehensively study the mechanism of resistance to F. oxysporum, transcriptome sequencings of root tissues from Lilium pumilum inoculated with F. oxysporum or sterile water for 6, 12, or 24 h were performed. A total of 50 GB of data were obtained from the transcriptome sequencings of the 6 L. pumilum samples, and 217 098 Unigenes were obtained after the de novo assembly, of which 38.36% Unigenes were annotated. The sequencing results showed that the numbers of differentially expressed genes at 6, 12, and 24 h after inoculation compared with the control were 111, 254, and 2500, respectively. The functional enrichment analysis of the differentially expressed genes showed that several pathways were involved in responses of L. pumilum, mainly including starch and sucrose metabolism, glycolysis/gluconeogenesis, phenylpropanoid biosynthesis, plant hormone signal transduction, flavonoid biosynthesis, vitamin B6 (VB6) biosynthesis, acid biosynthesis, proteasome, and ribosome. Transcription factor analysis revealed that the WRKY and ERF families played important roles in responses of L. pumilum to F. oxysporum. The results of this study elucidate the molecular responses to F. oxysporum in lily and lay a theoretical foundation for improving lily breeding and strategies for lily basal rot resistance. [ABSTRACT FROM AUTHOR]

Examination of the bulbs of Lilium pumilum (Liliaceae) led to the isolation of four novel steroidal glycosides (1–4) with a 2,3,4-trisubstituted β-d-glucopyranosyl unit. In 1 and 3, the α-L-arabinopyranosyl moiety is linked to C-3 of the inner trisubstituted β-D-glucopyranosyl group and is present as an usual 4C1 conformation. In contrast, in 2 and 4, the α-L-arabinopyranosyl moiety, which is attached to C-4 of the inner trisubstituted β-D-glucopyranosyl group, is present as a 1C4 conformation. The structures of the new steroidal glycosides were determined based on the results of spectroscopic analyses, including two-dimensional (2D) NMR data and hydrolysis.

• LpSOS1 , a gene encoding Na+/H+ antiporter, was isolated from Lilium pumilum and has been found to enhance salt stress tolerance in plants. • Overexpression of LpSOS1 in Arabidopsis enhanced tolerance to salt stress, as indicated by a lower malondialdehyde content; higher levels of the related antioxidant reductases superoxide dismutase, peroxidase, and catalase; and a decreased Na+/K+ ratio in transgenic plants. • Transgenic Arabidopsis plants expressing LpSOS1 demonstrated better growth and development under NaCl stress. • Transgenic lines overexpressing LpSOS1 showed much higher expression of salt stress-related genes under salt treatment compared to wild-type plants. Lilium pumilum has a strong salt tolerance. However, the molecular mechanism underlying its salt tolerance remains unexplored. Here, LpSOS1 was cloned from L. pumilum and found to be significantly enriched at high NaCl concentrations (100 mM). In tobacco epidermal cells, localization analysis showed that the LpSOS1 protein was primarily located in the plasma membrane. Overexpression of LpSOS1 resulted in up-regulation of salt stress tolerance in Arabidopsis , as indicated by reduced malondialdehyde levels and Na+/K+ ratio, and increased activity of antioxidant reductases (including superoxide dismutase, peroxidase, and catalase). Treatment with NaCl resulted in improved growth, as evidenced by increased biomass, root length, and lateral root growth, in both sos1 mutant (atsos1) and wild-type (WT) Arabidopsis plants that overexpressed LpSOS1 , Under NaCl treatment, atsos1 and WT Arabidopsis plants overexpressing LpSOS1 exhibited better growth, with higher biomass, root length, and lateral root quantity, whereas in the absence of LpSOS1 overexpression, the plants of both lines were wilted and chlorotic and even died under salt stress. When exposed to salt stress, the expression of stress-related genes was notably upregulated in the LpSOS1 overexpression line of Arabidopsis as compared to the WT. Our findings indicate that LpSOS1 enhances salt tolerance in plants by regulating ion homeostasis, reducing Na+/K+ ratio, thereby protecting the plasma membrane from oxidative damage caused by salt stress, and enhancing the activity of antioxidant enzymes. Therefore, the increased salt tolerance conferred by LpSOS1 in plants makes it a potential bioresource for breeding salt-tolerant crops. Further investigation into the mechanisms underlying lily's resistance to salt stress would be advantageous and could serve as a foundation for future molecular improvements. [ABSTRACT FROM AUTHOR]

• The salt tolerance threshold of lilium pumilum was preliminarily determined. • L. pumilum roots and leaves play a more important role in ion transport under salt stress. • Transcriptome analysis was performed on the leaves and roots of L. pumilum treated with salt stress. • The response mechanism of L. pumilum to salt stress is tissue-specific and involves different genes. • Plant hormone signal transduction, starch and sucrose metabolism, MAPK signaling pathway-plant, ion regulation and TFs are key pathways for L. pumilum to respond to salt stress. Salt stress seriously affects the growth and development of plants. Most species of lilies are not salt tolerant, thus salt affecting their geographical application. As a wild flower, Lilium pumilum is an excellent breeding material because of its bright colors, strong salt tolerance and cold tolerance. There are few studies on the salt tolerance mechanism and salt tolerance gene mining of L. pumilum. In this study, the salt tolerance threshold of L. pumilum was determined to be approximately 150 mM and the abundant ion translocation occurs in roots and leaves at the concentration. RNA-seq was used to characterize the transcriptome differences between L. pumilum leaves and roots treated with 150 mM NaCl for 2 and 12 h. By de novo assembly, 47,763 and 76,871 unigenes were identified in L. pumilum leaves and roots, respectively. Differentially expressed genes in leaves were higher than those in roots at 2 h, whereas the opposite result was obtained at 12 h. GO analysis of DEGs showed that transcriptional regulation and redox processes were significantly enriched, and KEGG annotation indicated that L. pumilum roots and leaves responded to salt stress mainly through phytohormone signaling, MAPK signaling pathways, and starch and sucrose metabolism. These results reveal important salt stress-related genes and metabolic pathways in L. pumilum , providing valuable information for exploring the salt tolerance mechanism of L. pumilum , screening salt tolerance genes and breeding new salt-tolerant L. pumilum varieties. [ABSTRACT FROM AUTHOR]

Lilium Pumilum with wide distribution is highly tolerant to salinity. The blue copper protein LpCPC ( Lilium pumilum Cucumber Peeling Cupredoxin ) gene was cloned from Lilium pumilum , which has the conserved regions of type I copper protein. Moreover, LpCPC has the closest relation to CPC from Actinidia chinensis using DNAMAN software and MEGA7 software. qRT-PCR indicated that LpCPC expression was higher in root and bulb of Lilium pumilum , and the expression of the LpCPC gene increased and reached the highest level at 12 h in bulbs under 20 mM NaHCO 3 . The transgenic yeast was more tolerant compared with the control under NaHCO 3 stress. Compared with the wild type, overexpressing plants indicated a relatively lower degree of wilting. In addition, the chlorophyll content, soluble phenol content, and lignin content of overexpressing lines were higher than that of wild-type, whereas the relative conductivity of overexpressing plants was significantly lower than that of wild-type plants. Expression of essential genes including NHX1 and SOS1 in salt stress response pathways are steadily higher in overexpression tobacco than that in wild-types. Transgenic lines had much higher levels of CCR1 and CAD , which are involved in lignin production, compared with wild-type lines. The yeast two-hybrid technique was applied to screen probable interacting proteins interacting with LpCPC. Eight proteins interacted with LpCPC were screened, and five of which were demonstrated to be associated with plant salinity resistance. Overall, the role of gene LpCPC is mediating molecule responses in increasing saline-alkali stress resistance, indicating that it is an essential gene to enhance salt tolerance in Lilium pumilum .

Alternative methods of in vitro cloning that involve both adventitious (direct) and callus intermediate (indirect) pathways were investigated for the endangered species Lilium pumilum. Plantlet regeneration was obtained from leaf explants, cultured on Murashige and Skoog (MS) basal medium supplemented with various combinations of auxins and cytokinins at different concentrations. About 30% of the explants directly formed adventitious shoots on MS medium containing 8.88 μM 6-benzyladenine (BA) and 2.69 μM α-naphthaleneacetic acid (NAA). For production of regenerable callus, callus formation followed by shoot induction was best when explants were initially cultured on MS medium supplemented with 9.05 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Regenerable calli were yellow or purple and readily regenerated shoots when subcultured onto MS medium containing 2.22 μM BA and 1.61 μM NAA. About 78% of the calli were able to produce adventitious shoots. Shoots were rooted on half-strength MS medium supplemented with 1.34 μM NAA and were successfully acclimatized to greenhouse conditions. This report describes an efficient method for the in vitro multiplication of whole plants from leaf explants of the endangered species L. pumilum. [ABSTRACT FROM AUTHOR]

Traditional Mongolian Medicine (TMM) plays an important role within the medical system of Mongolia nowadays. This medical system is rather unknown in the Western world, and detailed information can hardly be found in literature. In this article various aspects of TMM are highlighted. The eventful history of TMM is presented, and the centres which offer today’s traditional medical care are introduced. Institutions which provide education in TMM are outlined, and the latest developments in the national standard are highlighted, according to which the different institutions have to develop their curricula. Furthermore, an overview is given about herbal medicines in Mongolia and the health situation in this country. Finally, the international and Mongolian literature of Achillea asiatica, Dianthus versicolor, Euphorbia pallasii, Lilium pumilum, and Saussurea amara, which are all used in TMM to cure liver diseases, is reviewed. [ABSTRACT FROM AUTHOR]

Lilium pumilum is an important ornamental, culinary and medicinal bulbous plants with salt tolerance. However, salt tolerance of lily, particularly the bulb, has been studied relatively little, which brings challenges to the cultivation of lily varieties with high salt tolerance. Here, we performed transcriptome sequencing on the bulb organs of L. pumilum under salt stress treatment, analyzed differential gene expressed levels and then identified several key genes associated with salt stress tolerance at genome-wide scale. For the first time, we revealed the obvious response against salt stress for L. pumilum bulb organs, while distinct from those for root organs. Several key genes obtained through transcriptome analysis and DEG screening include NF-YB3 transcription factor, metallothionein type 2 protein, vicilin like seed storage protein and bidirectional sugar transporter SWEET14. Rather than typical ROS scavengers like superoxide dismutase, peroxidase, and glutathione transferase, non-typical ROS scavengers such as the metallothionein type 2 protein, and vicilin like seed storage protein were upregulated in our work. The bidirectional sugar transporter SWEET14 protein and the hormone signaling proteins such as E3-ubiquitin protein ligases, PYL4 and protein phosphatase 2C were also upregulated, suggesting the role of sugars and hormones in the bulb organ responses to salt stress. Co-expression analysis of the DEGs further confirmed that NF-YB3 transcription factor acted as a hub gene, suggesting that salt stress can promote flowering of L. pumilum . Taken together, we identified important candidate genes associated with salt tolerance of the L. pumilum bulb organs, which may provide the excellent basis for further in-depth salt tolerance mechanisms of the lily bulbs., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 So, Pak, Sun, Wang, Yan and Zhang.)

Lilium pumilum is an excellent wildflower germplasm resource with high resistance to salinity stress. The gene LpBCP plays an important role in salinity tolerance of L. pumilum. Studying the molecular mechanism of salinity resistance in L. pumilum will provide insights into multiple aspects, including breeding better varieties, environmental protection, improving soil conditions, etc. Conventional methods were used to determine different physiological indicators of Nicotiana benthamiana after NaHCO 3 treatment, i.e. chlorophyll content, soluble phenol content and lignin content. RT-qPCR was carried out to find expression of LpBCP in different organs and under abiotic stresses. DAB was used to detect H 2 O 2 in leaves in situ. A yeast two-hybrid system was used to screen for LpBCP interacting proteins. LpBCP was cloned from bulbs of L. pumilum. The highest expression of LpBCP was in roots and bulbs of transgenic plants. LpBCP-overexpressed plants showed less wilting, compared to WT plants. LpBCP transgenic plants have higher chlorophyll, soluble phenol and lignin content, and lower relative conductivity under 500 mM NaHCO 3 stress. In addition, H 2 O 2 scavenging in transgenic plants was much improved, indicating increased resistance to NaHCO 3 stress. Thirteen LpBCP-interacting proteins were screened using the yeast two-hybrid method and five were associated with salt stress. Based on our findings, LPBCP could be a key gene that can be used to improve L. pumilum salt tolerance., (© 2022 German Society for Plant Sciences, Royal Botanical Society of the Netherlands.)

Lilium pumilum is a perennial herb with ornamental edible and medicinal value. It is an excellent wild germplasm resource with wide distribution and strong resistance. The NAC family of transcription factors is unique to higher plants. The NAC family plays a regulatory role in plant growth and development and participates in plant responses to biotic and abiotic stresses. The LpNAC17 gene of L. pumilum was cloned and transformed into tobacco to investigate the response of transgenic tobacco to salt stress. The results showed that the net photosynthetic rate and contents of chlorophyll in LpNAC17 over-expressed tobacco were higher than those in the control plants, while the stomatal conductance, transpiration rate and intercellular CO 2 concentration were lower than those in the controls. The activity of superoxide dismutase, peroxidase, catalase, and the content of proline in LpNAC17 over-expressed tobacco were higher than those in the controls, while the content of malondialdehyde, superoxide anion, and hydrogen peroxide were lower than that in the control. Nitro-blue tetrazolium staining and 3,3'-diaminobenzidine tissue localization showed that the contents of O 2 - and H 2 O 2 in transgenic tobacco was lower than in the controls. The expression levels of NtSOD, NtPOD, NtCAT, NtHAK1, NtPMA4 , and NtSOS1 in the transgenic tobacco were higher than those in the controls. Therefore, this study provides a gene source for molecular breeding of salt-tolerant plants through genetic engineering, and lays a foundation for further research on salt-tolerant Lily., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Wang, Cui, Liu, Wang, Sun, Wang, Tan, Yan and Zhang.)

NAC transcription factors have multiple biological functions in plants. In this study, a new NAC transcription factor, LpNAC6 , was cloned from Lilium pumilum , and its salt and drought resistance functions were identified. We treated LpNAC6 transgenic tobacco plants with different intensities of alkali and drought stress. Results showed that LpNAC6 transgenic tobacco had enhanced alkali tolerance, but decreased drought tolerance. Antioxidant enzyme (SOD, POD, CAT) activity, chlorophyll content, proline content, and photosynthetic capacity of transgenic tobacco were significantly higher than those of wild-type tobacco, while the contents of MDA, H 2 O 2 , and O 2 − were significantly lower than those of wild-type tobacco. The expression level of stress-related genes in transgenic tobacco increased significantly, and the alkali tolerance was enhanced, but the opposite was true under drought stress. Our findings suggest that LpNAC6 has a reverse regulatory effect on alkaline and drought tolerance in plants, which is of great significance for plant screening and stress tolerance regulation of transgenic plants in arid saline–alkali land. • The function of a novel NAC transcription factor LpNAC6 cloned from Lilium pumilum was verified and analyzed. • LpNAC6 positively regulated plant responses to alkali and negatively regulated their responses to drought. • This study provides some new insights into the molecular mechanisms of alkali and drought resistance in the lily family. [ABSTRACT FROM AUTHOR]

Dianthus versicolor (Caryophyllaceae) and Lilium pumilum (Liliaceae) are two medicinal plants used in traditional Mongolian medicine to treat hepatic and gastrointestinal disorders. In this study aqueous (AE) and methanolic (ME) extracts of Dianthus versicolor and Lilium pumilum were investigated for their influence on the bile flow. The aqueous extracts of both plants were tested in absence and presence of 10μM taurocholic acid at three different concentrations (100, 250, and 500mg/L). The aqueous extract of Dianthus versicolor was further purified in order to locate the active principles. Two resulting fractions, one enriched in flavonoids and the other in sugars, were investigated for their influence on the bile flow in absence of taurocholic acid at 10, 20, and 40mg/L. The aqueous extracts of both plants were analysed qualitatively by LC-MSn and quantitatively by UV-spectrophotometry. Materials and methods: The bile flow experiments were performed in the isolated perfused rat liver. The compounds were identified by LC-DAD-MSn and TLC using references. The UV-spectrophotometric analysis was based on the monograph “Passiflorae herba” of the European Pharmacopoeia, and the total flavonoid contents were calculated and expressed as vitexin. Results: AE and ME of both plants increased the bile flow dose-dependently (between 9% and 30%), and no hepatotoxic effect was seen even during longer perfusions. Stimulation of bile secretion was comparable in the presence and in the absence of taurocholic acid. The flavonoid fraction of Dianthus versicolor increased the bile flow by 18% (p <0.05) at 40mg/L, which was comparable to the positive control cynarin. The phytochemical investigations of the Dianthus versicolor AE (total flavonoid content 1.78%) revealed the presence of the isovitexin derivative saponarin. In the AE of Lilium pumilum (total flavonoid content 1.04%) the flavonoids rutoside, kaempferol-3-O-rutinoside, and isorhamnetin-3-O-rutinoside were detected. Conclusions: The results show that choleresis under extract application is due to a stimulation of the bile-salt-independent bile flow which might be caused by the osmotic power of the extracts (hydrocholeresis). The flavonoids seem to contribute to the bile-flow-stimulating effect of Dianthus versicolor. Both plants exhibit a considerable choleretic effect that contributes to their use in traditional Mongolian medicine against gastrointestinal disorders. [Copyright &y& Elsevier]

Lilium pumilum DC. is a useful plant species not only for its showy flowers but also for its edible and medicinal values. Here we report on the complete chloroplast genome sequence of L. pumilum. The chloroplast genome is 152,573 bp in size and includes two inverted repeat regions of 52,984 bp, which is separated by a large single-copy region of 82,009 bp and a small single copy region of 17,580 bp. A total of 130 genes were predicted, including 38 tRNA, 8 rRNA, and 84 protein-coding genes. Phylogenetic analysis placed L. pumilum under the family Liliaceae. [ABSTRACT FROM AUTHOR]