Reproductive biology of an endangered lithophytic shrub and implications for its conservation.

BACKGROUND: Plants in cliff habitats may evolve specific reproductive strategies to cope with harsh environments, and unraveling these reproductive characteristics can improve our understanding of survival strategies and lithophyte evolution. This understanding is especially important for efforts to protect rare and endemic plants. Here, we investigated the reproductive biology of Lonicera oblata, an endangered lithophytic shrub that is scattered in highly fragmented and isolated cliff habitats of the Taihang and Yan mountains in North China. RESULTS: Flowers of L. oblata are herkogamous and protandrous, characteristics that can prevent autogamy at the single-flower level, and insects are necessary for pollination. The outcrossing index, pollen/ovule ratio, and the results of hand pollination were measured and all revealed a mixed mating system for L. oblata, that combines cross-fertilization and partial self-fertilization. The floral traits of L. oblata of zygomorphic and brightly yellowish corolla, heavy fragrance, and rich nectar, suggest an entomophilous pollination system. Sweat bees were observed as the most effective pollinators but their visiting frequencies were not high. Pollen limitation may limit the reproductive success of L. oblata. CONCLUSIONS: We determined the reproductive characteristics of L. oblata, a critically endangered species endemic to cliffs in North China, providing insight into its endangerment and suggesting conservation strategies. L. oblata has highly pollinator-dependent self-fertilization as part of a mixed mating system. Floral features such as low-flowering synchrony, asynchronous anthers dehiscence, and high duration of stigma receptivity, improve pollination efficiency in the case of low pollinator service. Our work provides reference information to understand the survival strategies and conservation of L. oblata and other lithophytes.

Cloning and expression of AGL19 gene in two Lonicera macranthoides varieties.

AGLl9 is an important regulator for flowering in plants and critical in controlling the morphogenesis of flower organs. The fulllength cDNAs of AGL19in conventional Lonicera macranthoides (Lm-AGL19) and the mutant 'Xianglei' cultivar (Lm-XL-AGL19) were obtained using rapid amplification of cDNA ends and the expression vectors for Lm-XL-AGL19were constructed to investigate the roles of AGL19 in the 'Xianglei' cultivar. Lm-AGL19 (GenBank: MK419948) and Lm-XL-AGL19 (GenBank: MK419948) were isolated from the conventional variety and 'Xianglei' cultivar of L. macranthoides, respectively. Lm-AGL19 is 1274 bp in length, whereas Lm-XL-AGL19 is 1264-bp long, and both include a 654 bp open reading frame, encoding 217 amino acids, which has a highly conserved MADS_MEF2_like domain and a moderately conserved K-box domain, belonging to the type II MADS-box family of genes. Quantitative real-time polymerase chain reaction indicated that the expression levels of these genes at different flowering stages were significantly different, and that the genes were also expressed in stems and leaves. Lm-XL-AGL19 is underexpressed at flowering period 5 that the key time node for corolla expansion and nonexpansion, while LM-AGL19 is overexpressed during this flowering period. AGL19 was speculated to be a functional gene causing different phenotypes in the two L. macranthoides varieties. The successfully constructed plant expression vector provides an experimental reference for further research on the function of this gene and the basis for the excellent phenotype of L. macranthoides 'Xianglei'.

Variations in morphology, physiology, and multiple bioactive constituents of Lonicerae Japonicae Flos under salt stress.

Lonicerae Japonicae Flos (LJF) is an important traditional Chinese medicine for the treatment of various ailments and plays a vital role in improving global human health. However, as unable to escape from adversity, the quality of sessile organisms is dramatically affected by salt stress. To systematically explore the quality formation of LJF in morphology, physiology, and bioactive constituents' response to multiple levels of salt stress, UFLC-QTRAP-MS/MS and multivariate statistical analysis were performed. Lonicera japonica Thunb. was planted in pots and placed in the field, then harvested after 35 days under salt stress. Indexes of growth, photosynthetic pigments, osmolytes, lipid peroxidation, and antioxidant enzymes were identified to evaluate the salt tolerance in LJF under different salt stresses (0, 100, 200, and 300 mM NaCl). Then, the total accumulation and dynamic variation of 47 bioactive constituents were quantitated. Finally, Partial least squares discrimination analysis and gray relational analysis were performed to systematically cluster, distinguish, and evaluate the samples, respectively. The results showed that 100 mM NaCl induced growth, photosynthetic, antioxidant activities, osmolytes, lipid peroxidation, and multiple bioactive constituents in LJF, which possessed the best quality. Additionally, a positive correlation was found between the accumulation of phenolic acids with antioxidant enzyme activity under salt stress, further confirming that phenolic acids could reduce oxidative damage. This study provides insight into the quality formation and valuable information to improve the LJF medicinal value under salt stress.

Integrated metabolic profiling and transcriptome analysis of pigment accumulation in Lonicera japonica flower petals during colour-transition.

BACKGROUND: Plants have remarkable diversity in petal colour through the biosynthesis and accumulation of various pigments. To better understand the mechanisms regulating petal pigmentation in Lonicera japonica, we used multiple approaches to investigate the changes in carotenoids, anthocyanins, endogenous hormones and gene expression dynamics during petal colour transitions, i.e., green bud petals (GB_Pe), white flower petals (WF_Pe) and yellow flower petals (YF_Pe). RESULTS: Metabolome analysis showed that YF_Pe contained a much higher content of carotenoids than GB_Pe and WF_Pe, with α-carotene, zeaxanthin, violaxanthin and γ-carotene identified as the major carotenoid compounds in YF_Pe. Comparative transcriptome analysis revealed that the key differentially expressed genes (DEGs) involved in carotenoid biosynthesis, such as phytoene synthase, phytoene desaturase and ζ-carotene desaturase, were significantly upregulated in YF_Pe. The results indicated that upregulated carotenoid concentrations and carotenoid biosynthesis-related genes predominantly promote colour transition. Meanwhile, two anthocyanins (pelargonidin and cyanidin) were significantly increased in YF_Pe, and the expression level of an anthocyanidin synthase gene was significantly upregulated, suggesting that anthocyanins may contribute to vivid yellow colour in YF_Pe. Furthermore, analyses of changes in indoleacetic acid, zeatin riboside, gibberellic acid, brassinosteroid (BR), methyl jasmonate and abscisic acid (ABA) levels indicated that colour transitions are regulated by endogenous hormones. The DEGs involved in the auxin, cytokinin, gibberellin, BR, jasmonic acid and ABA signalling pathways were enriched and associated with petal colour transitions. CONCLUSION: Our results provide global insight into the pigment accumulation and the regulatory mechanisms underlying petal colour transitions during the flower development process in L. japonica.

Comparative transcriptomics analysis revealing flower trichome development during flower development in two Lonicera japonica Thunb. cultivars using RNA-seq.

BACKGROUND: Lonicera japonica Thunb. (L. japonica) has the functions of clearing away heat and detoxifying, broad-spectrum antibacterial and anti-virus, etc. More than 70% of anti-inflammatory and cold Chinese patent medicines contain L. japonica. Trichomes comprise specialized multicellular structures that have the capacity to synthesize and secrete secondary metabolites and protect plants from biotic and abiotic stresses. The extraction of trichome secretions has great commercial value. However, little is known about the trichome formation mechanism in L. japonica. Therefore, the study of trichome development between different varieties provides a basis for selecting suitable planting resources. RESULTS: Here, we present a genome-wide comparative transcriptome analysis between two L. japonica cultivars, toward the identification of biological processes and functional gene activities that occur during flowering stage trichome development. In this study, the density and average lengths of flower trichomes were at their highest during three-green periods (S2). Using the Illumina RNA-Seq method, we obtained 134,304 unigenes, 33,733 of which were differentially expressed. In an analysis of 40 differentially expressed unigenes (DEGs) involved in trichome development, 29 of these were transcription factors. The DEGs analysis of plant hormone signal transduction indicated that plant growth and development may be independent of gibberellin (GA) and cytokinine (CTK) signaling pathways, and plant stress may be independent of jasmonic acid (JA) and ethylene (ET) signaling pathways. We screened several genes involved in the floral biosynthesis of odors, tastes, colors, and plant hormones, and proposed biosynthetic pathways for sesquiterpenoid, triterpenoid, monoterpenoid, flavonoid, and plant hormones. Furthermore, 82 DEGs were assigned to cell cycles and 2616 were predicted as plant resistance genes (PRGs). CONCLUSIONS: This study provides a comprehensive characterization of the expression profiles of flower development during the seven developmental stages of L. japonica, thereby offering valuable insights into the molecular networks that underly flower development in L. japonica.

Germination responses to winter warm spells and warming vary widely among woody plants in a temperate forest.

Winter underpins key ecological processes, such as dormancy loss and seedling emergence. Enhanced warm spells, together with warming are occurring and will continue in the future. The consequences of these climate phenomena on germination were investigated among co-occurring woody plants, whose seeds are bird-dispersed in autumn and require cold stratification for spring emergence. Seeds from nine common southeastern USA plants were collected in autumn. We verified that seeds of the study species required cold stratification for dormancy loss. We then examined the following aspects in the laboratory or field: effect of warm spells during cold stratification on germination, effect of a warm spell during winter on seed survival and germination phenology, and effect of warming from autumn dispersal through winter dormancy loss on timing of germination. While no consistent effects of warm spells were found in the laboratory on quantity of germination, warm spells advanced spring field germination for several species. Some species germinated during cold stratification and during warm spells, especially extreme spells, in the laboratory. In the field, about half of Lonicera maackii seedlings that emerged with a warm spell died by late winter. With warming from autumn through spring, laboratory germination shifted from spring to predominately autumn for some species. With precocious germination during warm spells or germination phenology shifts, two scenarios are possible. Seedlings may die during winter, reducing the size of the soil seed bank and number of emergents, or they would survive in warmer winters, which would give them a competitive advantage over spring-emerging seedlings.

Critical Phenological Events Affect Chemical Defense of Plant Tissues: Iridoid Glycosides in a Woody Shrub.

Plants are chemically-complex organisms; each individual contains diverse tissue-types, has the ability to differentially allocate secondary metabolites to these tissues and can change this allocation through time. The interaction of variation in chemical defense of different tissue types and variation in chemical defense through time, however, is rarely examined and has not been studied for iridoid glycoside-producing woody plants. In this study, we quantified allocation of iridoid glycosides (IGs) to the leaves, flowers, fruits, and seeds of 25 individuals of a long-lived shrub (Lonicera x bella Zabel, Caprifoliaceae), at five important phenological timepoints (leaf-out, flowering, fruit appearance, fruit ripening, and fruit dispersal) throughout a growing season. We found that leaves had 2x higher IG concentrations during flowering and fruiting than earlier in the season (after leaf-out), and later in the season (after fruit dispersal). The individual IG driving this increase in leaves during reproduction, secologanin, was also the most abundant IG in semiripe fruits. Flowers and seeds were composed of different proportions of individual IGs than fruits or leaves, but did not change across time and had overall low concentrations of IGs. In L. x bella, phenological events such as flowering and fruiting lead to an increase in leaf chemical defense that is likely to influence interactions with leaf-feeders. Our results stress the importance of considering phenology when sampling plants for the quantification of chemical defenses.

Transcriptional regulation of Lonicera japonica Thunb. during flower development as revealed by comprehensive analysis of transcription factors.

BACKGROUND: Lonicera japonica Thunb. flower has been used for the treatment of various diseases for a long time and attracted many studies on its potential effects. Transcription factors (TFs) regulate extensive biological processes during plant development. As the restricted reports of L. japonica on TFs, our work was carried out to better understand the TFs' regulatory roles under different developmental stages in L. japonica. RESULTS: In this study, 1316 TFs belonging to 52 families were identified from the transcriptomic data, and corresponding expression profiles during the L. japonica flower development were comprehensively analyzed. 917 (69.68%) TFs were differentially expressed. TFs in bHLH, ERF, MYB, bZIP, and NAC families exhibited obviously altered expression during flower growth. Based on the analysis of differentially expressed TFs (DETFs), TFs in MYB, WRKY, NAC and LSD families that involved in phenylpropanoids biosynthesis, senescence processes and antioxidant activity were detected. The expression of MYB114 exhibited a positive correlation with the contents of luteoloside; Positive correlation was observed among the expression of MYC12, chalcone synthase (CHS) and flavonol synthase (FLS), while negative correlation was observed between the expression of MYB44 and the synthases; The expression of LSD1 was highly correlated with the expression of SOD and the total antioxidant capacity, while the expression of LOL1 and LOL2 exhibited a negative correlation with them; Many TFs in NAC and WRKY families may be potentially involved in the senescence process regulated by hormones and reactive oxygen species (ROS). The expression of NAC19, NAC29, and NAC53 exhibited a positive correlation with the contents of ABA and H2O2, while the expression of WRKY53, WRKY54, and WRKY70 exhibited a negative correlation with the contents of JA, SA and ABA. CONCLUSIONS: Our study provided a comprehensive characterization of the expression profiles of TFs during the developmental stages of L. japonica. In addition, we detected the key TFs that may play significant roles in controlling active components biosynthesis, antioxidant activity and flower senescence in L. japonica, thereby providing valuable insights into the molecular networks underlying L. japonica flower development.

Discovery and mechanism study of a novel chromium-accumulating plant, Lonicera japonica Thunb.

Finding chromium-accumulating plants is of great interest for phytoremediation of soil contaminated by chromium (Cr). Inspired by Traditional Chinese Medicine, we examined the Cr-resistance and Cr-accumulation of Lonicera japonica Thunb. After a two-phase study using both soil and water culture, we found that L. japonica could be a novel Cr-accumulating plant, which contains an average Cr(III) content of 1297.14 mg.kg-1 in its leaves. The Cr enrichment factor and the Cr transport coefficient of Lonicera japonica was 5.19 and 1.79, respectively. Lonicera japonica is the fifth Cr-accumulating plant discovered worldwide, and the first Cr-accumulating woody plant ever discovered. The results support the conclusions drawn from studies of Cr-accumulating Leersia hexandra that oxalic acid production can increase Cr tolerance whereas citric acid or malic acid has no effect, suggesting that oxalic acid might be a common reason for Cr tolerance in all Cr-accumulating plants. Moreover, this study revealed that the production of anthocyanin and carotene can also increase Cr(III) tolerance, suggesting that anthocyanin and carotene might also account for Cr tolerance in Cr-accumulating plants. We believe that the discovery of Lonicera japonica as a Cr-accumulating plant will offer great opportunities in phytoremediation, and the success should be a strong sign that Traditional Chinese Medicine harbors more secrets to be uncovered with modern science.

Molecular cloning and spatiotemporal expression of APETALA1-like gene in Lonicera macranthoides.

APETALA1 (AP1), a floral meristem identity gene controls the flowering time and floral transition, and plays an important role in inflorescence and floral organ development. The full-length cDNA for AP1 was obtained by rapid amplification ofthe cDNA ends (RACE) so that the roles of AP1 in Lonicera macranthoides (Lm-AP1) could be better understood. AP1 (accession number in GenBank: MF418642) consisted of a 729-bp open reading frame encoding a protein that contained 242 amino acids, had a deduced molecular mass of 27.9919 kDa and a theoretical isoelectric point of 8.75. No signal peptide or transmembranedomains were detected in the sequences located in the nucleus, but it contained conserved sequences for MADS and the K-box. In the secondary structure, the alpha helix accounts for 60.74%, the beta turn 3.72%. The real-time polymerase chain reaction revealed that AP1 was more highly expressed in flowers, especially at the fourth flowering stage, which implied that it may play a role in flower development. Other L. macranthoides organs, such as stems and leaves, also expressed AP1. This research provided the basis for further analysis of the AP1 functional mechanism during L. macranthoides development.

Blue honeysuckle (Lonicera caerulea subsp. edulis (Turcz. ex Herder) Hultén.) berries and changes in their ingredients across different locations.

BACKGROUND: Interest in organic blue honeysuckle berries has increased in recent years. They are rich in various health-promoting compounds which are sensitive to different environmental factors and are modified during the growing season. RESULTS: Honeysuckle berries from different locations differed significantly in their contents of primary and secondary metabolites. The location Ogulin, with the highest altitude and consequently high UV radiation, had the highest phenolic content (259.85 mg per 100 g). Additionally, Vukovski Vrh, with the lowest temperature, had the highest ascorbic acid content (36.83 mg per 100 g), while Smartno pri Litiji and Visnja Gora, with the highest precipitation, had the highest organic acid contents (885.85 and 850.01 mg per 100 g respectively). A combination of stressful environmental conditions of temperature, water source and light intensity led to the highest saponin content in Visnja Gora (695 mg per 100 g), the highest tannin content in Dolnje Impolje (134 mg per 100 g) and the highest sugar content (2585.45 mg per 100 g) in Vucetinac. CONCLUSION: The contents of bioactive substances were influenced by various environmental factors such as temperature, UV radiation, altitude, light intensity and fruit ripening stage. Different compounds respond distinctly to different environmental factors. © 2017 Society of Chemical Industry.

Large-Scale Removal of Invasive Honeysuckle Decreases Mosquito and Avian Host Abundance.

Invasive species rank second only to habitat destruction as a threat to native biodiversity. One consequence of biological invasions is altered risk of exposure to infectious diseases in human and animal populations. The distribution and prevalence of mosquito-borne diseases depend on the complex interactions between the vector, the pathogen, and the human or wildlife reservoir host. These interactions are highly susceptible to disturbance by invasive species, including terrestrial plants. We conducted a 2-year field experiment using a Before-After/Control-Impact design to examine how removal of invasive Amur honeysuckle (Lonicera maackii) in a forest fragment embedded within a residential neighborhood affects the abundance of mosquitoes, including two of the most important vectors of West Nile virus, Culex pipiens and Cx. restuans. We also assessed any potential changes in avian communities and local microclimate associated with Amur honeysuckle removal. We found that (1) removal of Amur honeysuckle reduces the abundance of both vector and non-vector mosquito species that commonly feed on human hosts, (2) the abundance and composition of avian hosts is altered by honeysuckle removal, and (3) areas invaded with honeysuckle support local microclimates that are favorable to mosquito survival. Collectively, our investigations demonstrate the role of a highly invasive understory shrub in determining the abundance and distribution of mosquitoes and suggest potential mechanisms underlying this pattern. Our results also give rise to additional questions regarding the general impact of invasive plants on vector-borne diseases and the spatial scale at which removal of invasive plants may be utilized to effect disease control.

Rainfall variability counteracts N addition by promoting invasive Lonicera maackii and extending phenology in prairie.

Although encroaching woody plants have reduced the global extent of grasslands, continuing increases in soil nitrogen availability could slow this trend by favoring resident herbaceous species. At the same time, projected increases in rainfall variability could promote woody encroachment by aligning spatiotemporal patterns of soil moisture availability with the needs of woody species. We evaluated the responses of two deciduous woody species to these simulated environmental changes by planting seedlings of Quercus palustris and Lonicera maackii into tallgrass prairie communities grown under a factorial combination of increased rainfall variability and nitrogen addition. Lonicera maackii growth was reduced 20% by nitrogen addition, and increased rainfall variability led to 33% larger seedlings, despite greater competition for light and soil resources. In contrast, Q. palustris growth showed little response to either treatment. Increased rainfall variability allowed both species to retain their leaves for an additional 6.5 d in autumn, potentially in response to wetter end-of-season shallow soils. Our findings suggest increases in rainfall variability will counteract the inhibitory effect of nitrogen deposition on growth of L. maackii, extend autumn phenology, and promote the encroachment of some woody species into grasslands.

Degradation Dynamics and Dietary Risk Assessments of Two Neonicotinoid Insecticides during Lonicera japonica Planting, Drying, and Tea Brewing Processes.

The degradation dynamics and dietary risk assessments of thiamethoxam and thiacloprid during Lonicera japonica planting, drying, and tea brewing processes were systematically investigated using high-performance liquid chromatography. The half-lives of thiamethoxam and thiacloprid were 1.0-4.1 d in the honeysuckle flowers and leaves, with degradation rate constants k ranging from -0.169 to -0.696. The safety interval time was 7 d. The sun- and oven-drying (70 °C) percent digestions were 59.4-81.0% for the residues, which were higher than the shade- and oven-drying percentages at lower temperatures (30, 40, 50, and 60 °C, which ranged from 37.7% to 57.0%). The percent transfers of thiamethoxam and thiacloprid were 0-48.4% and 0-25.2%, respectively, for the different tea brewing conditions. On the basis of the results of this study, abiding by the safety interval time is important, and using reasonable drying methods and tea brewing conditions can reduce the transfer of thiamethoxam and thiacloprid to humans.

A comparative study of the dynamic accumulation of polyphenol components and the changes in their antioxidant activities in diploid and tetraploid Lonicera japonica.

Polyploidization is an effective method to achieve a higher yield of secondary metabolism active ingredients in medicinal plants. Polyphenols are the main active substances that contribute to the antioxidant activity of Lonicera japonica. For studying on the effect of chromosome doubling and harvest time on the dynamic accumulation of the main active substances and antioxidant capabilities of L. japonica, the polyphenol composition contents (7 phenolic acids and 3 flavonoids) and the antioxidant capacity in buds and flowers of diploid and tetraploid L. japonica at six different growth stages were determined by HPLC-DAD and three common antioxidant assays (FRAP, OH RSC and DPPH ARP), and the correlation between the dynamic accumulation of the polyphenol components and antioxidant capacity was also analyzed in current research. The results indicated that the content of the most determined phenolic acids and flavonoids and the antioxidant capacity in most of the growth stages from tetraploid plants were significantly higher than those in the diploid plants. Furthermore, the changes in the antioxidant activity presented a significant positive correlation with the variations in the chlorogenic acid, rutin, hyperoside, luteoloside in the two ploidy levels of L. japonica plants. The higher yields of chlorogenic acid (158.97, 164.00, 199.85 mg), luteoloside (5.44, 4.03, 6.31 mg), hyperoside (1.15, 1.06, 1.30 mg) and total flavonoids (9.87, 8.67, 11.10 mg) from 100 buds or flowers in tetraploid plants occurred during the S3-S5 stages, and these stages also exhibited higher antioxidant activities. Therefore, the stages of S3-S5 are recommended as the best time for harvesting high-yield, high-quality tetraploid Flos Lonicerae Japonicae.

Antioxidant capacities and anthocyanin characteristics of the black-red wild berries obtained in Northeast China.

Various edible berries widely accessible in nature in Northeast China are poorly exploited. The compositions and contents of anthocyanins in black (Padus maackii, Padus avium, Lonicera caerulea, and Ribes nigrum) and red (Ribes rubrum, Sambucus williamsii, Rubus idaeus, and Ribes procumbens) wild berries in Northeast China were firstly characterized by HPLC-DAD/ESI-MS(2). Twenty-three anthocyanins were detected and identified. Cyanidin glycosides were dominant in both berries. Six anthocyanins were reported for the first time in P. avium, R. rubrum, and Sambucus. Total anthocyanin content (TAC) ranged from 10mg/100gfreshweight (FW) (R. procumbens) to 1058mg/100gFW (P. maackii) among berries. The TACs and antioxidant activities assessed by DPPH and FRAP assays were much higher in black than in red berries. Black-red berries, especially P. maackii and P. avium, can be used in developing functional foods and in improving breeding programs.

Can arbuscular mycorrhizal fungi reduce Cd uptake and alleviate Cd toxicity of Lonicera japonica grown in Cd-added soils?

A greenhouse pot experiment was conducted to study the impact of arbuscular mycorrhizal fungi--Glomus versiforme (Gv) and Rhizophagus intraradices (Ri) on the growth, Cd uptake, antioxidant indices [glutathione reductase (GR), ascorbate peroxidase (APX), superoxide dismutase (SOD), catalase (CAT), ascorbate (ASA), glutathione (GSH) and malonaldehyde (MDA)] and phytochelatins (PCs) production of Lonicera japonica in Cd-amended soils. Gv and Ri significantly increased P acquisition, biomass of shoots and roots at all Cd treatments. Gv significantly decreased Cd concentrations in shoots and roots, and Ri also obviously reduced Cd concentrations in shoots but increased Cd concentrations in roots. Meanwhile, activities of CAT, APX and GR, and contents of ASA and PCs were remarkably higher in Gv/Ri-inoculated plants than those of uninoculated plants, but lower MDA and GSH contents in Gv/Ri-inoculated plants were found. In conclusion, Gv and Ri symbiosis alleviated Cd toxicity of L. japonica through the decline of shoot Cd concentrations and the improvement of P nutrition, PCs content and activities of GR, CAT, APX in inoculated plants, and then improved plant growth. The decrease of shoot Cd concentrations in L. japonica inoculated with Gv/Ri would provide a clue for safe production of this plant from Cd-contaminated soils.

[Study on influence factors of seed germination and seeding growth of Lonicera macranthoides].

In order to improve reproductive efficiency and quality standard, the influence factors of seed germination and seeding growth of Lonicera macranthoides werew studied. The fruit and seed morphological characteristics of L. macranthoides were observed, the seed water absorbing capacity was determined, and different wet sand stratification time, temperature and germination bed treatment were set up. The effects of the parameters on seed germination and seedling growth were analysed. There was no obstacles of water absorption on L. macranthoides seed, quantity for 22 h water absorption was close to saturation. In the first 80 d, with the increase of the stratification time, seed initial germination time was shortened, germination rate and germination potential was improved. Stratification for 100 d, germination rate decreased. At 15 ℃, seed germination and seedling growth indicators were the best. The seedling cotyledon width in light was significantly higher than that in dark. Seeds on the top of paper and top of sand germination rate, germination potential, and germination index was significantly higher than that of other germination bed and mildew rate is low. The optimal conditions of seeds germination test was stratified in 4 ℃ wet sand for 80 d, 15 ℃ illuminate culture on the top of paper or top of sand. The first seeding counting time was the 4th day after beginning the test, the final time was the 23th day. The germination potential statistical time was the 13th day after beginning the test.

Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters.

A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.

Screening and characterization of endophytic Bacillus and Paenibacillus strains from medicinal plant Lonicera japonica for use as potential plant growth promoters

Abstract A total of 48 endophytic bacteria were isolated from surface-sterilized tissues of the medicinal plant Lonicera japonica, which is grown in eastern China; six strains were selected for further study based on their potential ability to promote plant growth in vitro (siderophore and indoleacetic acid production). The bacteria were characterized by phylogenetically analyzing their 16S rRNA gene similarity, by examining their effect on the mycelial development of pathogenic fungi, by testing their potential plant growth-promoting characteristics, and by measuring wheat growth parameters after inoculation. Results showed that the number of endophytic bacteria in L. japonica varied among different tissues, but it remained relatively stable in the same tissues from four different plantation locations. Among the three endophytic strains, strains 122 and 124 both had high siderophore production, with the latter showing the highest phosphate solubilization activity (45.6 mg/L) and aminocyclopropane-1-carboxylic acid deaminase activity (47.3 nmol/mg/h). Strain 170 had the highest indoleacetic acid (IAA) production (49.2 mg/L) and cellulase and pectinase activities. After inoculation, most of the six selected isolates showed a strong capacity to promote wheat growth. Compared with the controls, the increase in the shoot length, root length, fresh weight, dry weight, and chlorophyll content was most remarkable in wheat seedlings inoculated with strain 130. The positive correlation between enzyme (cellulose and pectinase) activity and inhibition rate on Fusarium oxysporum, the IAA production, and the root length of wheat seedlings inoculated with each tested endophytic strain was significant in regression analysis. Deformity of pathogenic fungal mycelia was observed under a microscope after the interaction with the endophytic isolates. Such deformity may be directly related to the production of hydrolytic bacterial enzymes (cellulose and pectinase). The six endophytic bacterial strains were identified to be Paenibacillus and Bacillus strains based on the results of 16S rRNA gene sequencing analysis and their physiological and biochemical characteristics. Results indicate the promising application of endophytic bacteria to the biological control of pathogenic fungi and the improvement of wheat crop growth.