Genomic and functional characterization of coleopteran insect-specific α-amylase inhibitor gene from Amaranthus species.

The smallest 32 amino acid α-amylase inhibitor from Amaranthus hypochondriacus (AAI) is reported. The complete gene of pre-protein (AhAI) encoding a 26 amino acid (aa) signal peptide followed by the 43 aa region and the previously identified 32 aa peptide was cloned successfully. Three cysteine residues and one disulfide bond conserved within known α-amylase inhibitors were present in AhAI. Identical genomic and open reading frame was found to be present in close relatives of A. hypochondriacus namely Amaranthus paniculatus, Achyranthes aspera and Celosia argentea. Interestingly, the 3'UTR of AhAI varied in these species. The highest expression of AhAI was observed in A. hypochondriacus inflorescence; however, it was not detected in the seed. We hypothesized that the inhibitor expressed in leaves and inflorescence might be transported to the seeds. Sub-cellular localization studies clearly indicated the involvement of AhAI signal peptide in extracellular secretion. Full length rAhAI showed differential inhibition against α-amylases from human, insects, fungi and bacteria. Particularly, α-amylases from Helicoverpa armigera (Lepidoptera) were not inhibited by AhAI while Tribolium castaneum and Callosobruchus chinensis (Coleoptera) α-amylases were completely inhibited. Molecular docking of AhAI revealed tighter interactions with active site residues of T. castaneum α-amylase compared to C. chinensis α-amylase, which could be the rationale behind the disparity in their IC50. Normal growth, development and adult emergence of C. chinensis were hampered after feeding on rAhAI. Altogether, the ability of AhAI to affect the growth of C. chinensis demonstrated its potential as an efficient bio-control agent, especially against stored grain pests.

Isolation of Calenduloside E from Achyranthes bidentata Blume and its effects on LPS/D-GalN-induced acute liver injury in mice by regulating the AMPK-SIRT3 signaling pathway.

BACKGROUND: Acute liver injury (ALI) is a frequent fatal liver disease with a high mortality. Calenduloside E (CE) is a pentacyclic triterpenoid derived from Achyranthes bidentata Blume. It has been found that liver injury is associated with mitochondrial dysfunction, and activation of the AMPK-SIRT3 signaling pathway protects the mitochondrial function to play a role in resistance to the disease. However, whether CE is protective against ALI through the AMPK-SIRT3 signaling pathway is unclear. PURPOSE: To clarify the influences of Calenduloside E (CE) isolated from Achyranthes bidentata Blume on LPS/D-GalN-induced Acute liver injury (ALI). METHODS: A mouse model of ALI was developed, intraperitoneal injection of 10 µg/kg LPS and 700 mg/kg D-GalN, histopathological, oxidative stress, and immune inflammation of the mice were monitored. The mechanism of CE influencing liver injury was investigated by examining the gut microbiota, mitochondrial dysfunction, and the AMPK-SIRT3 signaling pathway. The antagonistic effects of specific AMPK and SIRT3 blocker, as well as AMPKα1, AMPKα2, SIRT3 transfection-mediated silencing were investigated to confirm the role of the AMPK-SIRT3 signaling pathway in this process. RESULTS: CE relieved liver pathological damage of mice and led to reduced oxidative stress and immune inflammation in mice, affected the balance of gut microbiota in mice with liver injury, as well as energy metabolism, and regulated mRNA and protein expressions of AMPK-SIRT3 signaling pathway. In addition, in vitro studies showed that CE relieved mitochondrial respiratory and protein expressions of AMPK-SIRT3 signaling pathway in LPS/D-GalN-induced AML12 and LX2 cells, and such effect was blocked by AMPK and SIRT3 inhibitors. Furthermore, silencing of AMPKα1, AMPKα2, and SIRT3 blocked the effects of CE. Overall, the influences of CE on mice with liver injury is tuned by the AMPK-SIRT3 signaling pathway. CONCLUSION: CE mediates mitochondrial function and eventually regulate energy metabolism by regulating the AMPK-SIRT3 signaling pathway. The results of this study provide molecular evidences for application of CE in treatment of ALI and provide references to the drug development for ALI.

Deuterium labeling for investigating de novo synthesis of terpene volatiles in Achyranthes bidentata.

PURPOSE OF WORK: Plants synthesize and accumulate secondary metabolites as defensive volatiles against diverse stresses. We aim to unravel the jasmonate-inducible volatile de novo synthetic metabolites in plants using a deuterium-labeling technique. Jasmonic acid and its methyl ester (MeJA) are well-documented for inducing defensive volatiles. Here, we have developed an efficient deuterium oxide (D2O)-based labeling approach to determine the extent of de novo synthetic metabolites in a model plant A. bidentata bidentata. The labeling approach was demonstrated on quantitative profiling of terpene volatile organic compounds (VOCs) elicited by airborne MeJA in Achyranthes plants. We show, for the first time that airborne MeJA-elicited terpene VOCs are predominantly and differentially de novo synthesized except for a homoterpene, (3E)-4,8-dimethyl-1,3,7-nonatriene, which is weakly and least labelled with deuterium. D2O is therefore an efficient labeling source for investigating de novo synthetic metabolites of terpene VOCs in planta.

Achyranthes bidentata polypeptides reduces oxidative stress and exerts protective effects against myocardial ischemic/reperfusion injury in rats.

Achyranthes bidentata, a Chinese medicinal herb, is reported to be neuroprotective. However, its role in cardioprotection remains largely unknown. Our present study aimed to investigate the effects of Achyranthes bidentata polypeptides (ABPP) preconditioning on myocardial ischemia/reperfusion (MI/R) injury and to test the possible mechanisms. Rats were treated with ABPP (10 mg/kg/d, i.p.) or saline once daily for one week. Afterward, all the animals were subjected to 30 min of myocardial ischemia followed by 4 h of reperfusion. ABPP preconditioning for one week significantly improved cardiac function following MI/R. Meanwhile, ABPP reduced infarct size, plasma creatine kinase (CK)/lactate dehydrogenase (LDH) activities and myocardial apoptosis at the end of reperfusion in rat hearts. Moreover, ABPP preconditioning significantly inhibited superoxide generation, gp91phox expression, malonaldialdehyde formation and enhanced superoxide dismutase activity in I/R hearts. Furthermore, ABPP treatment inhibited PTEN expression and increased Akt phosphorylation in I/R rat heart. PI3K inhibitor wortmannin blocked Akt activation, and abolished ABPP-stimulated anti-oxidant effect and cardioprotection. Our study demonstrated for the first time that ABPP reduces oxidative stress and exerts cardioprotection against MI/R injury in rats. Inhibition of PTEN and activation of Akt may contribute to the anti-oxidant capacity and cardioprotection of ABPP.

Achyranthes bidentate extracts protect the IL-1ß-induced osteoarthritis of SW1353 chondrocytes.

Osteoarthritis, the most common joint disease worldwide, is a degenerative disease characterized by cartilage degeneration and inflammation. The active ingredients in the traditional Chinese medicinal plant Achyranthes bidentate can be used to treat waist, leg, and joint pain caused by rheumatism arthralgia. In this study, we identified the optimal microwave extraction protocol for saponins from A. bidentate, evaluated their protective effects against IL-1ß-induced inflammation in SW1353 human chondrocytes, and explored their protective pathway. The microwave-extraction parameters required to obtain the maximum yield of A. bidentate saponins using 80% ethanol were identified using response surface methodology. The parameters were solid-liquid ratio, 1:10; extraction time, 20 min; power, 721 W; temperature, 65 °C. The actual yield of saponins extracted was to be 194.01 µg/mg extract. The SW1353 cells were pretreated with A. bidentate extract (ABE) at a concentration of 50 or 100 µg/mL for 3 h, after which an inflammatory response was stimulated using IL-1ß. The ABE significantly reduced the expression of proinflammatory factors IL-6, TNF-α, COX-2, iNOS, PGE2, and NO, and inhibited NF-κB activity, effectively attenuating the inflammatory response. ABE also inhibited MMP13 and ADAMTS-5 expression, reducing IL-1ß-induced degradation of the extrachondral matrix. This confirmed that ABE effectively inhibits NF-κB activity and reduces IL-1ß-induced inflammation, extracellular matrix degradation, and expression of apoptotic proteins Bax and caspase-3. Therefore, ABE has potential as a new botanical drug for preventing osteoarthritis.

[Achyranthes bidentata polysaccharide inhibits the adipogenic differentiation of rat bone marrow mesenchymal stem cells by blocking the PPARγ/TRPV4 pathway].

Objective To investigate the effect of Achyranthes bidentata polysaccharides (ABPS) on adipogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and its mechanism. Methods Five SD rats were sacrificed, and the BMSCs were dissected and isolated. The BMSCs were adherently cultured to logarithmic growth phase after identification, and treated with different doses of ABPS for 48 hours. The cell survival rates were detected by MTT assay. The highest dose of ABPS without toxicity to BMSCs was selected for subsequent experiments. Cells were randomly divided into control group, ABPS group, rosiglitazone group and ABPS combined with rosiglitazone group. Cell survival rates were detected by MTT assay. Triglyceride (TG) levels in BMSCs were detected by spectrophotometry. Lipid droplet formation in BMSCs was observed by oil red O staining. The mRNA and protein expression of peroxisome proliferater-activated receptor γ (PPARγ), transient receptor potential vanilloid 4 (TRPV4) and CCAAT/enhancer binding protein α (C/EBPα) were detected by real time quantitative PCR and Western blot analysis. Results The dose of ABPS≤200 mg/L had no obvious toxic effect on the growth of BMSCs after 48 hours, and the cell survival rate of 400 mg/L ABPS group was lower. Compared with the control group, the ABPS group showed decreased levels in TG, decreased relative expression of PPARγ, TRPV4 and C/EBPα mRNA and protein, and the decreased number of cytoplasmic lipid droplets. In the rosiglitazone group, observation reported the decreased cell survival rate, increased TG level, increased relative expression levels of PPARγ, TRPV4 and C/EBPα mRNA and protein, along with the increased number of cytoplasmic lipid droplets. Compared with the ABPS group, the cell survival rate was decreased, TG level was increased, the relative expression levels of PPARγ, TRPV4 and C/EBPα mRNA and protein increased, and the number of cytoplasmic lipid droplets increased in the ABPS combined with rosiglitazone group. Compared with rosiglitazone group, the survival rate was increased, TG level was decreased, the relative expression levels of PPARγ, TRPV4 and C/EBPα mRNA and protein were decreased, and the number of cytoplasmic lipid droplets was decreased in the ABPS combined with rosiglitazone group. Conclusion ABPS can inhibit adipogenic differentiation of BMSCs, and the mechanism may be related to the regulation of PPARγ/TRPV4 pathway.

Structural characterization and in vitro osteogenic activity of ABPB-4, a heteropolysaccharide from the rhizome of Achyranthes bidentata.

Achyranthes bidentata is a species of flowering plant that is mainly distributed in China. The A. bidentata rhizome is a famous traditional Chinese medicine that has been widely used to treat lumbago, arthritis, and bone hyperplasia. In this work, A. bidentata rhizome was isolated and purified to obtain a pectic polysaccharide (ABPB-4). Chemical and spectral analyses showed that ABPB-4 had a main chain of →4)-α-d-GalpA-(1→ and →2,4)-α-l-Rhap-(1→, and the branch chains included →4)-ß-d-Galp-(1→, →6)-ß-d-Galp-(1→, →3,6)-ß-d-Galp-(1→, →5)-α-l-Araf-(1→ and →3,5)-α-l-Araf-(1→, and it was terminated with α-l-Araf-(1→ and ß-d-Galp-(1→. At concentrations of 0.01, 0.02, and 0.04 µmol/L, ABPB-4 significantly promotes the proliferation, differentiation, and mineralization of MC3T3-E1 cells in vitro, and it appreciably enhances the mRNA expression levels of osteogenic-related genes in these cells. Overall, the results reported herein indicate that ABPB-4 has outstanding osteogenic activity, and that it may be used as an anti-osteoporosis agent in the future.

Distribution and biosynthesis of 20-hydroxyecdysone in plants of Achyranthes japonica Nakai.

There is increasing interest in phytoecdysteroids (PEs) because of their potential role in plant defense against insects. To understand the mechanism regulating their levels in plants, the fluctuation, distribution, and biosynthesis of PE 20-hydroxyecdysone (20E) examined in Achyranthes japonica. The total amount of 20E per individual plant initially remained at a constant level, and increased markedly after the first leaf pair (LP) stage, while the concentration of 20E in a given plant decreased rapidly during vegetative growth. In addition, the incorporation of [2-(14)C]-mevalonic acid into 20E did not differ significantly depending on plant organs and developmental stages, suggesting that biosynthesis of 20E is not restricted to particular organs or growth stages.

Metabolomic Investigation on Fermentation Products of Achyranthes japonica Nakai by Lactobacillus plantarum.

Fermentation has recently re-emerged as an approach for improved functionality of food products in addition to the traditional roles such as shelf life, taste, and texture. Here, we report dynamic changes in the metabolite profiles of Achyranthes japonica Nakai by Lactobacillus plantarum fermentation, primarily, the significant increases in representative functional ingredients, 20-hydroxyecdysone and 25S-inokosterone. Additionally, untargeted metabolite profiling showed 58% of metabolites underwent significant alteration. The most dynamic change was observed in cellobiose, which showed a 56-fold increase. Others were sugar alcohols and amino acids, while lyxitol and erythritol that were among the most dynamically down-regulated.

NAA and 6-BA promote accumulation of oleanolic acid by JA regulation in Achyranthes bidentata Bl.

Application of plant growth regulators has become one of the most important means of improving yield and quality of medicinal plants. To understand the molecular basis of phytohormone-regulated oleanolic acid metabolism, RNA-seq was used to analyze global gene expression in Achyranthes bidentata treated with 2.0 mg/L 1-naphthaleneacetic acid (NAA) and 1.0 mg/L 6-benzyladenine (6-BA). Compared with untreated controls, the expression levels of 20,896 genes were significantly altered with phytohormone treatment. We found that 13071 (62.5%) unigenes were up-regulated, and a lot of differentially expressed genes involved in hormone or terpenoid biosynthesis, or transcription factors were significantly up-regulated. These results suggest that oleanolic acid biosynthesis induced by NAA and 6-BA occurs due to the expression of key genes involved in jasmonic acid signal transduction. This study is the first to analyze the production and hormonal regulation of medicinal A. bidentata metabolites at the molecular level. The results herein contribute to a better understanding of the regulation of oleanane-type triterpenoid saponins accumulation and define strategies to improve the yield of these useful metabolites.

Accumulation and dynamic trends of triterpenoid saponin in vegetative organs of Achyranthus bidentata.

The relationship between structural features of various vegetative organs and triterpenoid saponin accumulation in Achyranthus bidentata Blume was investigated using anatomy, histochemistry and phytochemistry. The results showed that the primary and secondary structures of roots, and the structures of stems and leaves of A. bidentata, were similar to those of ordinary dicotyledonous plants. The enlargement of its roots, however, was primarily associated with growth and differentiation of tertiary structures. There were collateral medullary vascular bundles in addition to the normal vascular bundles in the stem. The tertiary structure was not only main parts in the roots of A. bidentata, but also important storage region of triterpenoid saponin in its growth and development. The stem may be the essential transport organ of triterpenoid saponin, while palisade parenchyma may be the primary synthesis location. In November, the total quantity of triterpenoid saponin and overall biomass in the roots reach a maximum level. This was the best time, therefore, to harvest the roots and corresponded to the traditional harvest period. Despite the withered appearance of leaves, stems also contained substantial amounts of triterpenoid saponin, and it was recommended that the stems of A. bidentata should be used.

[Effect of phosphorus on copper tolerance in Achyranthes bidentata].

OBJECTIVE: To study the effect of phosphorus on copper tolerance in Achyranthes bidentata. METHOD: A PVC pipe experiment was conducted to study the interactive effects of phosphorus (P) and copper (Cu), on growth, elemental accumulation and chemical constituents of A. bidentata. Two levels of elemental P were applied at 0 (P0) and 100 ( P100) mg x kg(-1) soil with 5 levels of Cu at 0 (Cu0), 100 (Cu100), 200 (Cu200), 200 (Cu400), 200 (Cu600) mg x kg(-1) soil, respectively. RESULT AND CONCLUSION: The biomass production between different Cu treatments, phosphorus treatment showed significant differences. The biomass reached the maximum value as the concentration of Cu and P was 100 mg x kg(-1). Low concentration of Cu improved the growth of A. bidentata. The growth was blocked as Cu concentration reached 200 mg x kg(-1) in soil, however the contents of oleanolic acid and ecdysterone in roots of A. bidentata had not influenced by Cu. P could improved the copper tolerance in A. bidentata and increased root yield. The Cu concentration in soil of the cultivation bases must be below 200 mg x kg(-1) in order to produce good quality of medicinal material.

Chemical and metabolic analysis of Achyranthes bidentate saponins with intestinal microflora-mediated biotransformation by ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry coupled with metabolism platform.

Achyranthes bidentate (AB) is a typical traditional Chinese medicine (TCM) that has been widely used in clinical practices for more than a thousand years. Modern pharmacological studies have shown that triterpene saponins are the main pharmacological active ingredients in AB. Meanwhile, the poor oral bioavailability of triterpene saponins in AB indicates that these ingredients are probably metabolized by intestinal microflora before absorption. In this work, an integrated analysis based on ultra-performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MS) combined with a metabolism platform was developed to identify the chemical constituents and intestinal metabolic profiles of triterpene saponins in AB. As a result, a total of 40 triterpene saponins (including thirty-eight oleanane-type, one hederagenin-type and one machaerinate-type triterpene saponin) were identified from the AB extract. Moreover, 39 biotransformation products mediated by intestinal microflora were characterized, which mainly underwent four metabolic reactions including deglycosylation, glycosylation, oxidation and dehydrogenation. To our knowledge, the in vitro metabolites of AB through intestinal microflora metabolism, especially triterpene saponins, have not been studied previously. The obtained results could be helpful for the further evaluation of the pharmacokinetics and the pharmacological activity of triterpene saponins of AB in vivo.

Achyranthes bidentata polypeptides promotes migration of Schwann cells via NOX4/DUOX2-dependent ROS production in rats.

Achyranthes bidentata polypeptides (ABPP), an active polypeptides isolated from the aqueous extract of Achyranthes bidentata Blume, contributes to the regeneration of injured peripheral nerves by promoting migration of Schwann cells (SCs). In this study, we aimed to investigate the possible mechanism underlying the ABPP-induced migration of primary cultured rat SCs. Transwell migration assays indicated that ABPP promoted SCs migration in a concentration-dependent manner by inducing production of NADPH-oxidase (NOX)-derived reactive oxygen species (ROS). Inhibition of ROS production by NOXs inhibitor apocynin (APO) or diphenyleneiodonium (DPI) partially blocked ABPP-mediated SCs migration. Furthermore, by using real-time polymerase chain reaction analysis and siRNA interference technique, we verified the participation of NOX subunit 4 (NOX4) and dual oxidase 2 (DUOX2) in ABPP-induced ROS production and consequential SCs migration. Taken together, these results demonstrated that ABPP promoted SCs migration via NOX4/DUOX2-activated ROS in SCs.

Achyranthes bidentata polypeptide k improves long-term neurological outcomes through reducing downstream microvascular thrombosis in experimental ischemic stroke.

Achyranthes bidentata Bl. (A. bidentata) occupies an important position in traditional Chinese medicine owing to the property of promoting the circulation of blood and removing stasis. Achyranthes bidentata polypeptide k (ABPPk) is one of the active components isolated from A. bidentata. We previously demonstrated that ABPPk has potent neuroprotective effects against neuronal apoptosis both in vitro and in vivo, but the roles and mechanisms of ABPPk on long-term functional recovery after ischemic stroke remain unknown. In the current study, we investigated the neuroprotective effects of ABPPk on filament transient middle cerebral artery occlusion (tMCAO) rats and found that ABPPk reduced the infarct volume and maintained the neuronal integrity in the ischemic penumbra. Moreover, we found that ABPPk might reduce the formation of downstream microthrombus through preventing ischemic-induced oxidative damage of brain endothelial cells and activation of tissue factor (TF), plasminogen activator inhibitor-1 (PAI-1), and NF-κB. ABPPk also inhibited polymorphonuclear leukocytes (PMNs) infiltration and matrix metalloproteinase-2/-9 (MMP-2/-9) activation in the ischemic penumbra. Morris water maze, foot fault test, and modified neurological severity score were assessed for a period of 6 weeks following tMCAO. ABPPk improved long-term recognition abilities and neurological outcomes after stroke compared with saline-treated rats. Taken together, these results suggested that ABPPk is beneficial to the improvement of long-term outcomes after transient cerebral ischemia injury and can be used as a potential neuroprotective agent.

Interplant communication: airborne methyl jasmonate is essentially converted into JA and JA-Ile activating jasmonate signaling pathway and VOCs emission.

Methyl jasmonate (MeJA) was identified as an airborne signal involved in mediating interplant defense response communications over a decade ago. However, how MeJA activates plant defense systems and what becomes of the compound after it has done so has, thus far, remained unknown. To investigate this, Achyranthes bidentata plants were exposed to deuterated methyl jasmonate (d(2)MeJA) followed by absolute quantification of metabolic products of d(2)MeJA, and emissions of volatile organic compound (VOC) as defensive markers. We found that d(2)MeJA was metabolized mainly into deuterated jasmonic acid (d(2)JA) and jasmonoyl isoleucine (d(2)JA-Ile), and to a much lesser extent, deuterated jasmonoyl leucine (d(2)JA-Leu). Increases in d(2)JA-Ile/Leu and also endogenous JA-Ile/Leu were tightly co-related with, and significantly influenced the pattern and amount of, VOC emissions. The amount of accumulated d(2)JA-IIe was 13.1-fold higher than d(2)JA-Leu, whereas the amounts of JA-IIe and JA-Leu accumulated were almost identical. This study demonstrates that exogenous MeJA activates defensive systems (such as VOC emissions) in receiver plants by essentially converting itself into JA and JA-IIe and initiating a signal transduction leading to VOC emissions and induction of endogenous JA-IIe and JA-Leu, which in turn cause further amplification of VOC emissions.

Supercritical fluid extraction of ecdysterone from the roots of Achyranthes bidentata BL.

Ecdysterone has been found in a great many plants and animals and has some valuable pharmaceutical properties. The present study was conducted to investigate optimal conditions for the extraction of the compound by supercritical fluid extraction from the roots of Achyranthes bidentata BL. An orthogonal array design (OAD), OA(9)(3(4)), was employed as a chemometric method for optimization of the extraction of ecdysterone from the herbal medicine. Four parameters, namely, pressure and temperature of the supercritical fluid, the dynamic extraction time, and the flow rate of dimethyl sulfoxide, were studied and optimized by a three-level OAD. Determinations of the extracts were performed by high-performance liquid chromatography. The effects of the parameters were studied using analysis of variance. The results shown that the yield of ecdysterone could be influenced by the four parameters to a similar degree. The yield for DMSO-modified supercritical CO(2) was in the range from 0.65 to 1.03 mg/g under the selected conditions. In comparison with methanol-modified supercritical CO(2 )and Soxhlet extraction, a higher yield was obtained when DMSO-modified supercritical CO(2) was used.

[Primary study on absorption, translocation and accumulation of N, P and K of Achyranthes bidentata].

OBJECTIVE: To study the absorption, translocation and accumulation of N, P and K on Achyranthes bidentata. METHOD: The contents of N, P and K were determined by mean of sulfuric acid-hydrogen peroxide assimilating method, vanadium-ammonium molybdate colorimetric method and flame photometric method, respectively. RESULT: The contents of N, P and K in the plant were decreasing during the growth period. The absorption rates of the three nutrients by A. Bidentata showed double-peak curves in the whole growth period, maximum absorption rate appeared in the middle ten days of October. About 8.59 kg of N, 1.36 kg of P and 7.40 kg of K were needed to produce each 100 kg root. CONCLUSION: The key nutrients absorption period is in the first ten days of September and in the middle ten days of October.

[Primary study on suspension cell culture and polysaccharides content of Achyranthes bidentata].

OBJECTIVE: To study the relationship between the condition of cell suspension culture and the polysaccharides content in Achyranthes bidentata. METHOD: The methods of orthogonal test and alone factor test were used to study the effect of culture time, inoculation concentration, carbon source and pH. RESULT AND CONCLUSION: The results showed that the optimum medium to induce polysaccharides in A. bidentata was 1/4MS + 6-BA 0.5 + 2,4-D 1.0 mg x L(-1) + sucrose 2% + glucose 2% + fructose 1% cultured after 50 d. The suitable pH was 6.0, and the optimum inoculation concentration was 7g x L(-1).

[Comparative research on vitality and characteristics of seeds of Achyranthes bidentata in different producing area].

OBJECTIVE: To study the germination property of the seeds of Achyranthes bidentata from different producing area, providing foundation for choosing the better seed resources. METHOD: The germination rate, germination energy, germination index, vigor and the characteristics of the seeds of A. bidentata from different producing area were compared, and the relative electric conductivity was measured. The characteristics of the seed were recorded through the Anymicro DSS YT-5M microscope digital camera, and the differences were analyzed. RESULT: Seed from Henan Boai and Hebei Anguo showed the better germination rate, germination energy, germination index and vigor. CONCLUSION: The seeds from both Henan Boai and Hebei Anguo showed the better quality.