N-Exponential Fertilization Could Affect the Growth and Nitrogen Accumulation of <i>Metasequoia glyptostroboides</i> Seedling in a Greenhouse Environment

Metasequoia glyptostroboides (M. glyptostroboides) is a unique plant species related to relic flora in China. It plays a positive role in afforestation and its long-term protection with high paleoclimate research value. However, due to the nutrients-supply deficiency, it is a big challenge to cultivate the high-quality seedlings of M. glyptostroboides. In this study, a pot experiment in a greenhouse environment was carried out to identify the effect of N-exponential fertilization on the growth and nutrient distribution of M. glyptostroboides seedling. The M. glyptostroboides rooted seedlings with 12-month growth were chosen. Different N fertilizer levels with conventional fertilization (CF: 5.0 g seedling⁻¹), exponential fertilization including EF1, EF2, EF3 and EF4 were determined. The relevant growth indexes were measured after 210-day growth. The results indicated that non-significant differences in seedlings’ height and ground diameter were found among the above treatments (P > 0.05); At the same time, N-exponential fertilization promoted the M. glyptostroboides’s biomass in different organs (P < 0.05), with the maximum total biomass under EF3 treatment. The N accumulation in root and stem of the N-exponential fertilization treatments were increased in to some extent (P < 0.05). The maximum N accumulation was also found under EF3 treatment. Therefore, steady-state nutrition and superior growth performance of M. glyptostroboides could be obtained by N-exponential fertilization of 5.0 g cutting⁻¹.     

Effects of Nano-Cerium Oxide on Seed Germination and Seedling Growth of <i>Vitex negundo</i>

Cerium oxide nanoparticles (CeO₂NPs) are likely to have dramatic impacts on plant performances, yet the effects of CeO₂NPs on seed germination and seedling growth have not been fully explored. In this study, the seed germination and seedling growth of subshrub species Vitex negundo under different concentrations of CeO₂NPs (low-1 mg/L, medium-100 mg/L, high-500 mg/L) have been discussed. Results showed that: (1) The seed germination rate reduces by 11.25% and 2.5% under the low and medium concentrations of CeO₂NPs, respectively, but increased by 7.08% under the high concentration; (2) CeO₂NPs had significant effects on the growth traits (root length, shoot height and biomass) of seedlings, being the highest under the medium concentration and the lowest under the highest concentration; (3) The superoxide dismutase activity was the maximum (355.91 U/g), but the protein concentration was the minimum (3.85 ug/mL) under the high concentration of CeO₂NPs. Our results indicated that the effects of CeO₂NPs on seed germination and seedling growth are concentrationdependency, i.e., low and medium concentrations inhibited while high concentration promoted seed germination, however, seedling growth showed opposite responses. Therefore, appropriate CeO₂NPs concentrations are beneficial to the seed germination and seedling growth of Vitex negundo and improve the physiological performance of seedlings and enhance their adaptability to environmental adversity.     

AM Fungi and <i>Piriformospora indica</i> Improve Plant Growth of <i>Pinus elliottii</i> Seedlings

Pinus elliottii is an exotic afforestation pine extensively distributed in southern parts of China. In order to understand whether endophytic fungi can affect seedling growth of P. elliottii, Piriformospora indica (Pi), Funnelifcrmis mosseae (Fm), and Diversispora tortuosa (Dt) were inoculated respectively, and the non-inoculated group was set as control. The growth indexes, the contents of soluble sugar and soluble protein, and plant endogenous hormone levels in the leaves of P. elliottii, were analyzed. The results showed that Fm, Dt and Pi colonized the P. elliottii roots to form mycorrhizal structure and chlamydospores arranged in beads respectively. Three fungal inoculants exhibited the stimulated growth responses, whilst Dt illustrated the most positive effect on plant height, single fresh weight, trunk diameter and root system structure, compared with the control. On the other hand, the soluble sugar and soluble protein contents were increased distinctively in mycorrhizal plants. The endogenous IAA, GA3, ZR contents were increased, while the ABA contents were reduced in mycorrhizal plants versus non-mycorrhizal plants. The fungi-induced endogenous hormone changes triggered plant growth improvement of P. elliottii seedlings. This research unraveled the positive effect of AM fungi and P. indica on growth of pine seedlings, while, more application of endophytic fungi to fields needs to be explored.     

Mycorrhizal Networks Interacting with Litter Improves Nutrients and Growth for One Plant through the Vary of N/P Ratio under Karst Soil

Arbuscular mycorrhizae (AM) fungi affect nutrient uptake for host plants, while it is unclear how AM fungi interacting with soil litter affect plant growth and nutrient utilization through mycorrhizal networks in karst soil of deficient nutrients beyond the rhizosphere. An experiment was conducted in a microcosm composed of a planting compartment for Cinnamomum camphora seedlings with or without Glomus mosseae fungus (M⁺ vs. M⁻ ) and an adjacent litter compartment containing or not containing additional litter material of Arthraxon hispidus (L⁺ vs. L⁻ ), where the compartments are connected either by nylon mesh of 20 μm or 0.45 μm which either allow available mycorrhizal networks within the litter compartment or prevent mycelium entering into the litter compartment (N⁺ vs. N⁻ ). Plant biomass and nutrients were measured. The results showed that the addition of litter changed the symbiotic process in mycorrhizal colonization, spore, and hyphal density, which when in association with the host plant then affected the biomass, and accumulations of N (nitrogen) and P (phosphorus) in the individual plant as well as root, stem, and leaf respectively. AM fungi increased N and P accumulations and N/P ratio in individual plants and plant tissues. A decrease of the N/P ratio of the individual plant was observed when AM fungus interacted significantly with litter through mycorrhizal networks in the litter compartment. The results indicate that the C. camphora seedlings benefited from litter in nutrient utilization of N and P through the vary of N/P ratio when accessing mycorrhizal networks. These findings suggest that mycorrhizal networks interacting with litter improve growth and nutrients of N and P for plants through the vary of N/P ratio in order to alleviate nutrient limitation under karst soil.     

Biomass and nutrient dynamics of Chinese fir seedlings under conventional and exponential fertilization regimes

Containerized Chinese fir (Cunninghamia lanceolata (Lamb) Hook) were reared from seed at four fertilizer levels (0, 15, 45, 75 mg N seedling-1 season-1) and two topdressing schedules (conventional or exponential) for a 22-week greenhouse rotation to assess growth, nutrition and nutrient loading capacity of seedlings. Extra P supplemented high fertilization (or nutrient loading) treatments to test for induced deficiency of this element. The schedule and rate of fertilization significantly affected growth and nutrient dynamics of the seedlings. Steady-state nutrition and superior growth performance were achieved by seedlings fertilized exponentially at the operational dose (15 mg N), yielding 23, 72 and 52% more in respective biomass, N uptake and P uptake than seedlings fertilized conventionally at the equivalent dose. The improved growth and fertilizer efficiency were attributed to close synchronization of exponential nutrient supply with exponential growth and nutrient demand of plants. High dose exponential fertilization (45 and 75 mg N) induced steady state-nutrition late in the season, increasing seedling N and P uptake by 72–83% and 50–96% compared to low dose exponential fertilization, demonstrating effective nutrient loading of plants without changing biomass. The extra P stimulated P uptake without altering growth or N uptake, thus P was probably not limiting during the greenhouse culture despite high N additions.     

Antibacterial Activity and Mechanisms of Ethanol Extracts from <i>Scutellaria baicalensis</i> Georgi and <i>Magnolia officinalis</i> against <i>Phytophthora nicotianae</i>

Phytophthora nicotianae causes substantial economic losses in most countries where tobacco is produced. At present, the control of P. nicotianae mainly depends on chemical methods, with considerable environmental and health issues. We investigated the effects of ethanol extracts from Scutellaria baicalensis Georgi (SBG) and Magnolia officinalis (MO). On mycelial growth, sporangium formation, and zoospore release of P. nicotianae. Both extracts inhibited the growth of P. nicotianae, with mycelial growth inhibition rates of 88.92% and 93.92%, respectively, at 40 mg/mL, and EC50 values of 5.39 and 5.74 mg/mL, respectively. The underlying mechanisms were the inhibition of sporangium formation, the reduction of zoospore number, and the destruction of the mycelium structure. At an SBG extract concentration of 16.17 mg/mL, the inhibition rates for sporangia and zoospores were 98.66% and 99.39%, respectively. At an MO extract concentration of 2.87 mg/mL, the production of sporangia and zoospores was completely inhibited. The hyphae treated with the two plant extracts showed different degrees of deformation and damage. Hyphae treated with SBG extract showed adhesion and local swelling, whereas treatment with MO extract resulted in broken hyphae. Mixture of the extracts resulted in a good synergistic effect.     

Evaluation of Pre-Emergence and Post-Emergence Herbicides for Weed Management in <i>Miscanthus sacchariflorus</i> and <i>Miscanthus sinensis</i>

Miscanthus, is a promising bioenergy crop, considered superior to other bioenergy crops because of its higher water and nutrient use efficiency, cold tolerance, and higher production of biomass. Broadleaf weeds and grass weeds, cause major problems in the Miscanthus field. A field experiment was conducted in 2018 and 2019, to assess the effects of pre-emergence (alachlor and napropamide) and post-emergence herbicides (nicosulfuron, dicamba, bentazon, and glufosinate ammonium) on broadleaf and grass weeds in M. sinensis and M. sacchariflorus fields. The weed control efficiency and phytotoxicity of pre- and post-emergence herbicides were evaluated at 30 days after treatment (DAT) and compared to those of the control plots. The results showed wide variations in the susceptibility of the weed species to the treated herbicides. Treatment with nicosulfuron 40 g.a.i.ha⁻¹ provided the most effective overall weed control (with 10% visual injury), without affecting the height and biomass of neither Miscanthus species in the field. Post-emergence herbicides such as glufosinate ammonium 400 g.a.i.ha⁻¹ and dicamba 482 g.a.i.ha⁻¹ were effective and inhibited the growth and density of the majority of weeds to a 100%; however, they showed significant phytotoxicity (toxicity scale of 1–10) to both species of Miscanthus. The application of glufosinate ammonium caused severe injuries to the foliar region (90% visual injury) of both Miscanthus sps. Comparatively, M. sinensis showed a slightly higher tolerance to the herbicides nicosulfuron, bentazon and napropamide with 10% visual injury at the recommended dose than M. sacchariflorus. The present study clearly showed that infestation of broadleaf and grass weeds in Miscanthus fields can cause significant damage to the growth and biomass of Miscanthus and applying pre-emergence and post-emergence herbicides effectively controls the high infestation of these weeds.     

Effects of Different Geographical Aspects and Ontogenetic Variability on Total Hypericin Content of <i>Hypericum triquetrifolium</i> Turra. and <i>Hypericum scabrum</i> L.

The present study was conducted to determine the total hypericin contents of Hypericum triquetrifolium Turra. and Hypericum scabrum L. species which are naturally distributed in the flora of Siirt province, Turkey. Hypericin contents of Hypericum species grown in different geographical aspects (North, South, East, and West), and it was measured at different harvest times (full blooming and post blooming period). In the current study, it has been determined that total hypericin content varies considerably according to aspects, plant developmental stages (ontogenetic variance), and species. According to species x aspect interaction, the highest total hypericin content was recorded from the west aspect (3.13 mg/g) in Hypericum triquetrifolium, while, the lowest hypericin content was also obtained from the west aspect (1.22 mg/g) in Hypericum scabrum. When the highest total hypericin content was analyzed according to aspect x species x harvest time interaction, the highest total hypericin content was produced from Hypericum triquetrifolium at the harvest of west aspect with 5.28 mg/g, while the minimum amount of hypericin was obtained from the same aspect in Hypericum scabrum with 0.50 mg/g. In species x harvest time interaction, the highest total hypericin content was obtained from the full bloom (3.10 mg/g) harvest in Hypericum triquetrifolium, while the lowest hypericin was obtained from the full bloom (1.26 mg/g) harvest in Hypericum scabrum. The data suggest that the average total hypericin content was 2.26 mg/g in Hypericum triquetrifolium and 1.28 mg/g in Hypericum scabrum.     

Differential Responses of <i>NHX1</i> and <i>SOS1</i> Gene Expressions to Salinity in two <i>Miscanthus sinensis</i> Anderss. Accessions with Different Salt Tolerance

The lignocellulosic crop Miscanthus spp. has been identified as a good candidate for biomass production. The responses of Miscanthus sinensis Anderss. to salinity were studied to satisfy the needs for high yields in marginal areas and to avoid competition with food production. The results indicated that the relative advantages of the tolerant accession over the sensitive one under saline conditions were associated with restricted Na⁺ accumulation in shoots. Seedlings of two accessions (salt-tolerant ‘JM0119’ and salt-sensitive ‘JM0099’) were subjected to 0 (control), 100, 200, and 300 mM NaCl stress to better understand the salt-induced biochemical responses of genes involved in Na⁺ accumulation in M. sinensis. The adaptation responses of genes encoding for Na⁺ /H⁺ antiporters, NHX1 and SOS1 to NaCl stress were examined in JM0119 and JM0099.The cDNA sequences of genes examined were highly conserved among the relatives of M. sinensis based on the sequencing on approximate 600 bp-long cDNA fragments obtained from degenerate PCR. These salt-induced variations of gene expression investigated by quantitative real-time PCR provided evidences for insights of the molecular mechanisms of salt tolerance in M. sinensis. The expression of NHX1 was up-regulated by salt stress in JM0119 shoot and root tissues. However, it was hardly affected in JM0099 shoot tissue except for a significant increase at the 100 mM salt treatment, and it was salt-suppressed in the JM0099 root tissue. In the root tissue, the expression of SOS1 was induced by the high salt treatment in JM0119 but repressed by all salt treatments in JM0099. Thus, the remarkably higher expression of NHX1 and SOS1 were associated with the resistance to Na⁺ toxicity by regulation of the Na⁺ influx, efflux, and sequestration under different salt conditions.     

氮素指数施肥对杉木无性系苗生长及养分含量的影响

以1年生无性系杉木幼苗为材料,采用温室盆栽方法,设定对照、常规施肥和3个指数施肥共5个处理,施氮总量分别为每株0、0.5、0.5、1.0、2.0 g,施氮20次,间隔为10 d,施氮210 d后,测定杉木的苗高、地径、生物量、叶绿素荧光参数及养分含量.结果表明: 施肥显著促进了杉木无性系的苗高、地径和生物量的增长,以施氮量1.0 g·株^-1的指数施肥处理最佳,分别为59.0 cm、6.0 mm、52.99 g·株^-1;杉木无性系苗木叶绿素荧光参数特征均表现为指数施肥显著大于常规施肥,其中指数施肥的光化学淬灭系数、表观电子传递速率和PSⅡ的有效量子产量均随施氮总量的增加呈先增加后减小的趋势,在施氮量为1.0 g·株^-1时达到最佳,分别为0.89、35.79和0.71.杉木无性系苗木氮含量表现为叶>根>茎.与对照相比,常规施肥根、茎、叶的氮含量分别增加39.6%、16.6%和41.1%,指数施肥根、茎、叶的氮含量分别增加22.6%～81.4%、27.3%～152.6%和73.6%～135.5%.施氮量为1.0和2.0 g·株^-1的指数施肥处理杉木幼苗根、茎和叶氮含量均显著大于常规施肥,而不同器官磷含量和钾含量无显著差异.从苗木生长性状、叶绿素荧光参数特征、植株氮磷钾含量等综合考虑,在本试验条件下,施氮量为1.0 g·株^-1的指数施肥是杉木无系性苗木温室培育的适宜方法.     

Participation of Auxin Transport in the Early Response of the <i>Arabidopsis</i> Root System to Inoculation with <i>Azospirillum brasilense</i>

The potential of Plant Growth Promoting Rhizobacteria (PGPR) has been demonstrated in the case of plant inoculation with bacteria of the genus Azospirillum which improves yield. A. brasilense produces a wide variety of molecules, including the natural auxin indole-3-acetic acid (IAA), as well as other phytoregulators. However, several studies have suggested that auxin induces changes in plant development during their interaction with the bacteria. The effects of A. brasilense Sp245 on the development of Arabidopsis thaliana root were investigated to help explain the molecular basis of the interaction. The results obtained showed a decrease in primary root length from the first day and remained so throughout the exposure, accompanied by a stimulation of initiation and maturation of lateral root primordia and an increase of lateral roots. An enhanced auxin response was evident in the vascular tissue and lateral root meristems of inoculated plants. However, after five days of bacterization, the response disappeared in the primary root meristems. The role of polar auxin transport (PAT) in auxins relocation involved the PGP1, AXR4-1, and BEN2 proteins, which apparently mediated A. brasilense-induced root branching of Arabidopsis seedlings.     

不同指数施肥方法下长白落叶松播种苗的需肥规律

为了探讨长白落叶松苗木的最适N含量和供N速率,应用3种不同指数施肥方法在大田条件下进行试验。结果表明:经过111d的生长,供N总量为30mg.株-1的指数施肥处理下的苗木可以获得相对较高的生物量积累和N含量水平,该处理还可以使得苗木在103d的生长中获得0.29 m.g株-.1d-1的N添加速率。在该速率下,苗木生物量、N含量和N吸收效率分别可以达到637.80 m.g株-1、8.10 m.g株-1和41.77%。根据生物量和N含量随着供N量增加的二项式回归拟合结果,在111d的生长过程中24.3—33.7m.g株-1的供N总量可以使得苗木获得较高的生物量积累水平和N利用效率。试验设置的总供N量为30 m.g株-1的处理优于60 m.g株-1或70 m.g株-1。     

Characterization and Expression of Ammonium Transporter in Peach (<i>Prunus persica</i>) and Regulation Analysis in Response to External Ammonium Supply

As the preferred nitrogen (N) source, ammonium (NH₄⁺ ) contributes to plant growth and development and fruit quality. In plants, NH₄⁺ uptake is facilitated by a family of NH₄⁺ transporters (AMT). However, the molecular mechanisms and functional characteristics of the AMT genes in peach have not been mentioned yet. In this present study, excess NH₄⁺ stress severely hindered shoot growth and root elongation, accompanied with reduced mineral accumulation, decreased leaf chlorophyll concentration, and stunned photosynthetic performance. In addition, we identified 14 putative AMT genes in peach (PpeAMT). Expression analysis showed that PpeAMT genes were differently expressed in peach leaves, stems and roots, and were distinctly regulated by external NH₄⁺ supplies. Putative cis-elements involved in abiotic stress adaption, Ca²⁺ response, light and circadian rhythms regulation, and seed development were observed in the promoters of the PpeAMT family genes. Phosphorylation analysis of residues within the C-terminal of PpeAMT proteins revealed many conserved phosphorylation residues in both the AMT1 and AMT2 subfamily members, which could potentially play roles in controlling the NH₄⁺ transport activities. This study provides gene resources to study the biological function of AMT proteins in peach, and reveals molecular basis for NH₄⁺ uptake and N nutrition mechanisms of fruit trees.     

Growth and nitrogen nutrition of Chinese fir seedlings exposed to nutrient loading and fertilization

Nutrient loaded and non-loaded Chinese fir (Cunninghamia lanceolata (Lamb) Hook) seedlings were transplanted in a pot trial to examine effects of exponential nutrient loading and fertilization treatments on first season growth and N nutrition. The treatments tested four rates of N (0, 30, 60, and 90 mg tree^-1) as a mixed NPK fertilizer applied before planting to create a soil fertility gradient, and two topdressings applied only to non-loaded seedlings later in the season. Nutrient loading alone consistently enhanced seedling growth on the four soil fertility classes, increasing respective biomass and N uptake 42, 45, 20 and 8%, and 65, 67, 29 and 18% more than non-loaded seedlings. The positive response was attributed to increased N retranslocation from higher nutrient reserves built up by loading during nursery culture. Net retranslocation from old shoots to new growth was highest soon after planting when nutrient stress was most severe. Pre-plant soil fertilization and post-plant topdressings were also effective in promoting seedling productivity, but equivalent additions yielded less biomass than that from nutrient loading alone. Implications are that exponential nutrient loading may be more efficient in improving early growth performance of Chinese fir seedlings than traditional field fertilization practices at plantation establishment, and may on competitive sites avoid problems of stimulating surrounding vegetation rather than trees. This revised version was published online in June 2006 with corrections to the Cover Date.     

Rapid Propagation of <i>Rhynchostylis retusa in Vitro</i>

An efficient regeneration system of Rhynchostylis retusa was established to provide technical reference for the application of tissue culture tube seedlings in production. The mixtures of callus and protocorm from aseptic germination were used as explants. The optimal media of each stage was selected for callus proliferation, protocorm occurrence and growth, rejuvenation and rooting via a single, complete combination and orthogonal experiment. The results showed that the optimal medium for callus proliferation, protocorms occurrence and growth was 1/2 Murashige and Skoog (MS) medium adding 50 g·L⁻¹ banana puree, 0.1 mg·L⁻¹ α-naphthaleneacetic acid (NAA), 1.5 mg·L⁻¹ 6-benzylaminopurine (6-BA) and 1.0 mg·L⁻¹ kinetin (KT) with 17.33 proliferation coefficient of callus and 19.63 occurrence coefficient of buds after 90 days. Then the buds occurred from protocorm were cultured on 1/2 MS medium including 100 g·L⁻¹ banana puree, 1.0 mg·L⁻¹ NAA, 2.0 mg·L⁻¹ 6-BA and 0.05 mg·L⁻¹ KT, in which the proliferation coefficient of callus was 10.32 and occurrence coefficient of buds reached 17.87. In the further subculture, the same medium was simultaneously used for callus proliferation, protocorm occurrence and bud growth. The plantlets developed roots in 1/2 MS medium containing 70 mL·L⁻¹ coconut water and 1.5 mg·L⁻¹ NAA with 100% rooting rates after 90 days. The survival rate was more than 90% after domestication and transplantation. This regeneration protocol will provide technique foundation for protecting wild resource and developing artificial cultivation.     

不同形态氮素施肥对小黑杨幼苗生长的影响

以小黑杨当年播种苗为材料,研究了不同施氮量（12,24和48 mg·株^-1）和不同形态氮素（有机氮和无机氮）施肥对小黑杨幼苗生长的影响,以探讨小黑杨对氮基酸类有机氮素施肥的生长响应。结果表明：无论施无机氮（硝酸铵）还是有机氮（精氨酸）,小黑杨幼苗的苗高、地径、总生物量都是随施氮量的增加而增加,但是中等施氮量处理的幼苗氮利用效率最高。无机氮（硝酸铵）和有机氮（精氨酸）处理的幼苗在相同施氮水平下生长表现无明显差异,施用有机氮可以与施用无机氮获得相同的促进苗木生长的效果。不同氨基酸及其组合肥料施用对小黑杨幼苗生长的影响显著。单一氨基酸施肥情况下,施用精氨酸促进苗木生长的效果最好,谷氨酸次之,甘氨酸最差;氨基酸组合施肥情况下,有精氨酸的组合施肥苗木生长好,有甘氨酸的组合施肥苗木生长差。不同氨基酸施肥处理对小黑杨幼苗各器官氮含量没有明显影响。     

Genome-Wide Analysis of the <i>KANADI</i> Gene Family and Its Expression Patterns under Different Nitrogen Concentrations Treatments in <i>Populus trichocarpa</i>

KANADI (KAN) is a plant-specific gene that controlled the polarity development of lateral organs. It mainly acted on the abaxial characteristics of plants to make the lateral organs asymmetrical. However, it had been less identified in woody plants. In this study, the members of the KAN gene family in Populus trichocarpa were identified and analyzed using the bioinformatics method. The results showed that a total of 8 KAN family members were screened out, and each member contained the unique GARP domain and conserved region of the family proteins. Phylogenetic analysis and their gene structures revealed that all KAN genes from P. trichocarpa, Arabidopsis thaliana, and Nicotiana benthamiana could be divided into four subgroups, while the eight genes in P. trichocarpa were classified into three subgroups, respectively. The analysis of tissue-specific expression indicated that PtKAN1 was highly expressed in young leaves, PtKAN6 was highly expressed in young leaves and mature leaves, PtKAN2, PtKAN5, and PtKAN7 were highly expressed in nodes and internodes, PtKAN8 was highly expressed in roots, and PtKAN3 and PtKAN4 showed low expression levels in all tissues. Among them, PtKAN2 and PtKAN6, and PtKAN4 and PtKAN5 might have functional redundancy. Under high nitrogen concentrations, PtKAN2 and PtKAN8 were highly expressed in mature stems and leaves, respectively, while PtKAN4, PtKAN5, and PtKAN7 were highly expressed in roots. This study laid a theoretical foundation for further study of the KAN gene-mediated nitrogen effect on root development.     

Mycorrhiza and seedling establishment in a subarctic meadow: Effects of fertilization and defoliation

Abstract. Question: How does changing resource availability induced by fertilization and defoliation affect seedling establishment and mycorrhizal symbiosis in a subarctic meadow? Location: 610 m a.s.l., Kilpisjärvi (69°03’N, 20°50’E), Finland. Methods: A short‐term full‐factorial experiment was established, with fertilization and defoliation of natural established vegetation as treatments. Seeds of two perennial herbs Solidago virgaurea and Gnaphalium norvegicum were sown in natural vegetation and their germination and growth followed. At the final harvest we measured the response in terms of arbuscular mycorrhizal (AM) colonization, biomass and nitrogen concentration of the seedlings and the established vegetation. Results: Germination rate was negatively affected by defoliation in the unfertilized plots. The shoot biomass of S. virgaurea seedlings was reduced by the defoliation and fertilization treatments, but not affected by their interaction. In G. norvegicum, the germination rate and the seedling shoot biomass were negatively correlated with moss biomass in the plots. In the established plants the arbuscular colonization rate was low and defoliation and fertilization treatments either increased or did not affect the colonization by AM fungi. In the seedlings, the colonization rate by AM fungi was high, but it was not affected by treatments. Both seedlings and established plants were colonized by dark septate fungi. Conclusions: Reduction of plant biomass by herbivores can have different effects on seedling growth in areas of high and low soil nutrient availability. The weak response of AM colonization to defoliation and fertilization suggests that AM symbiosis is not affected by altering plant resource availability under the conditions employed in this study.     

<i>In vitro</i> Germination and Micropropagation of <i>Aconitum vilmorinianum</i>: An Important Medicinal Plant in China

Aconitum vilmorinianum, a well-known traditional Chinese herb, is recently being threatened by overexploitation and environment disturbance. This study was conducted to provide propagation methods through in vitro germination and explant cultivation. Germination was stimulated up to 66.00% on Murashige and Skoog (MS) medium containing 2.0 mg L⁻¹ 6-benzylaminopurine (BAP), 0.1 mg L⁻¹ 1-napthaleneacetic acid (NAA), and 30 g L⁻¹ sucrose. Three bacteria (Pantoea agglomerans, Erwinia persicina, and Pseudomonas tolaasii) would be responsible for consistent contamination during germination. The latter two were effectively eradicated after disinfected. The influence of explant types and hormone combinations on direct and indirect organogenesis was evaluated in the present work. The frequency of shoot induction from axillary bud explants was 100% on the MS fortified with 2.0 mg L⁻¹ BAP and 0.3 mg L⁻¹ NAA. Shoots multiplication was optimized on MS medium supplemented with 0.1 mg L⁻¹ thidiazuron (TDZ) and 0.1 mg L⁻¹ NAA. High callus induction percentage (96.67%) was obtained from stem segments on MS medium with 2.0 mg L⁻¹ 2,4-D, then successfully regenerated into shoots on MS medium in the presence of 0.1 mg L⁻¹ TDZ and 0.2 mg L⁻¹ NAA. The present work could be useful for the utilization and conservation of this valuable species.