In this research,we explored the effect of three groups of water treatments,including severe drought(the corresponding water content of cultivated substrate 5%-10%),moderate drought(45%-50%) and control(85%-90%),and different drought stress time(15,30,45 d) on the glandular trichome density(TD),stomatal density(SD) and volatile exudates of Schizonepeta tenuifolia.The results showed that there were two kinds of glandular trichomes on the surface of S. tenuifolia leaves: peltate and capitate glandular trichomes. The density of capitate glandular trichomes(CTD) was higher than that of peltate glandular trichomes(PTD). Both CTD and PTD on the abaxial surface of leaf were higher than those on the adaxial surface. Under severe drought stress,the CTD and SD were higher than the other two treatments. Under the same stress time,the biomass and leaf surface area of S. tenuifolia decreased with the deepening of stress degree. As the stress time prolonged,the surface area of leaves and biomass gradually increased,and the TD and SD decreased. The most abundant compound in volatile exudates of S. tenuifolia was pulegone. Under drought stress,the relative content of pulegone decreased,and the relative content of other monoterpenoids such as D-limonene and menthone increased. The n-hexadeconic acid and 2-methyl-1-hexadecanol were detected only at the stress of 15 d,while menthone was detected at the stress of 30 d and45 d. Drought stress affected the leaf growth and secondary metabolism of S. tenuifolia.
Droughts ; Lamiaceae ; Plant Exudates ; Plant Leaves ; Trichomes

The quality of moxa is a key factor affecting the efficacy of moxibustion. Traditional moxa grades are evaluated by the leaf-to-moxa ratio, but there is a lack of support from scientific data. Scanning electron microscopy(SEM), Image Pro Plus, Van Soest method, and stimultaneous thermal analysis(TGA/DSC) were used to characterize the scientific implication of the combustion differences between moxa with different leaf-to-moxa ratios(processed by crusher). The results showed that the median lengths from non-secretory trichomes(NSTs) of natural NSTs and moxa with leaf-to-moxa ratios of 3∶1, 5∶1, 10∶1, and 15∶1 were 542.46, 303.24, 291.18, 220.69, and 170.61 μm, respectively. The cellulose content of moxa increased significantly(P<0.05) with the increase in leaf-to-moxa ratio and the combustion parameters(T_i, t_i, D_i, C,-R_p,-R_v, S, D_b, and J_(total)) all showed an increasing trend. The correlation results showed that the burning properties of moxa(T_i,-R_v, t_i, and J_2) were significantly and positively correlated with cellulose content. NSTs with a length of 1-200 μm were significantly and positively correlated with J_2. NSTs with a length of 200-600 μm were significantly and positively correlated with J_1, T_(peak2), T_(peak1), and-R_v, and negatively correlated with J_(total), T_b, and t_b. As the leaf-to-moxa ratio increases, the NSTs in the moxa become shorter and the cellulose content increases, which is more conducive to ignition performance, heat release, and a milder, longer-lasting burn. The "NSTs-cellulose-TGA/DSC" quantitative evaluation method scientifically reveals the scientific connotation of the combustion of moxa with different leaf-to-moxa ratios and provides a scientific basis for the establishment of quality evaluation methods for moxa with different leaf-to-moxa ratios.
Trichomes ; Moxibustion ; Hot Temperature ; Plant Leaves

The basic features of glandular and non-glandular trichomes on leaves of Artemisia argyi( germplasms from Qichun,Ningbo,Tangyin,and Anguo,respectively) and related species A. stolonifera were observed by scanning electron microscopy( SEM)and compared. There were significant differences in trichome characteristics of leaves at all parts of A. argyi and A. stolonifera,which were closely related to the difference in chemical components. The length of non-glandular trichomes and size of glandular trichomes on middle leaves were the stablest. A. argyi and A. stolonifera can be distinguished by the density of glandular trichome. Additionally,the four germplasms of A. argyi can be discriminated via the density and curvature of non-glandular trichome. The density of non-glandular trichomes was the highest in A. stolonifera. For A. argyi,the germplasm from Qichun had the highest density of non-glandular trichomes on the abaxial surfaces of upper leaves and that from Ningbo had the largest non-glandular trichome curvature. With regard to the germplasm from Anguo,the T-shaped non-glandular trichomes of long stalks on the adaxial surfaces of the middle leaves were lodging-susceptible,and those with slender heads were wave-like. Statistics results of A. argyi and A. stolonifera are as follows: largest glandular trichomes on the adaxial and abaxial surfaces and highest glandular trichome density on the abaxial surfaces of the lower leaves in A. argyi germplasm from Ningbo,highest density of non-glandular trichomes on the abaxial surfaces of upper leaves in A. stolonifera,and highest density of glandular trichomes and non-glandular trichomes on the adaxial surfaces of the upper leaves in A. argyi germplasm from Qichun. According to the observation result under fluorescence microscope( FM),flavonoids were closely related to the size and density of non-glandular trichomes and size of glandular trichomes. The fluorescence intensity was the strongest and fluorescence area was the largest for flavonoids in A. argyi germplasms from Qichun and Tangyin,while the fluorescence for flavonoids was the weakest in A. stolonifera. It was the first time to observe and analyze the trichome ultrastructure of A. argyi leaves at different positions by SEM and FM. This study clarifies the differences between A. stolonifera and four famous A. argyi germplasms,which provides new evidence for the microscopic identification of A. argyi and its related species and serves as a reference for the study of the relationship of A. argyi structure with its components and functions.
Artemisia ; Flavonoids ; Microscopy, Electron, Scanning ; Plant Leaves ; Trichomes

Plant trichomes are special structures that originate from epidermal outgrowths. Trichomes play an important role in plant defense against pests and diseases, and possess economic and medicinal values. Study on molecular mechanism of plant trichomes will contribute to the molecular design breeding and genetic improvement of crops. In recent years, the regulation mechanism of trichome development has been basically clarified in the model plant Arabidopsis thaliana, while great progresses are also found in other plant species. In this review, we focus on the developmental regulation of trichome formation from gene and phytohormones levels in Arabidopsis and cotton (with unicellular trichomes), as well as in tomato and Artemisia annua (with multicellular trichomes). The research progress associated with trichomes is also introduced in other typical monocotyledons and dicotyledons. Finally, the research and application of plant trichomes are prospected.
Arabidopsis/genetics* ; Gene Expression Regulation, Plant ; Gossypium/genetics* ; Lycopersicon esculentum ; Plant Growth Regulators/metabolism* ; Trichomes/genetics*

Artemisia stolonifera is a relative of A. argyi. The two species are difficult to be distinguished due to the similarity in leaf shape and have even less distinctive features after processing. This study aims to establish a method to quickly distinguish between them. At the same time, we examined the reasonability and applicability of the specific polymerase chain reaction(PCR) method. The C/T single nucleotide polymorphism was detected at the position 202 of the sequence, based on which specific primers were designed to identify these two species. The PCR with the specific primer JNC-F and the universal primer ITS3R produced a specific band at 218 bp for A. argyi and no band for A. stolonifera, which can be used to detect at least 3% of A. argyi samples mixed in A. stolonifera samples. The PCR with the specific primer KY-F and the universal primer ITS3R produced a specific band at 218 bp for A. stolonifera and no band for A. argyi, which can be used to detect at least 5% of A. stolonifera samples mixed with A. argyi. The limit of detection of the established method was 5 ng DNA. The established PCR method can accurately distinguish between A. stolonifera and A. argyi, which provides an experimental basis for the quality control of A. stolonifera and determines whether the herbs are adulterated.
Artemisia/genetics* ; Trichomes ; Polymerase Chain Reaction ; Nucleic Acid Amplification Techniques ; Plant Leaves/genetics*

Hd-Zip, a unique transcription factor in plant kingdom, influences the growth, development, and secondary metabolism of plants. Hd-zip Ⅳ is thought to play an important role in trichome development of Schizonepeta tenuifolia. This study aims to explore the functions of StHD1 and StHD8 in Hd-zip Ⅳ subfamily in peltate glandular trichome development. To be specific, the expression patterns of the two genes and interaction between the proteins encoded by them were analyzed based on transcriptome sequencing and two-hybrid screening. The subcellular localization was performed and functions of the genes were verified in tobacco and S. tenuifolia. The results showed that StHD1 and StHD8 had high similarity to HD-Zip Ⅳ proteins of other plants and they all had the characteristic conserved domains of HD-Zip Ⅳ subfamily. They were located in the nucleus. The two genes mainly expressed in young tissues and spikes, and StHD1 and StHD8 proteins interacted with each other. The density and length of glandular trichomes increased significantly in tobacco plants with the overexpression of StHD1 and StHD8. Inhibiting the expression of StHD1 and StHD8 by VIGS(virus-induced gene silencing) in S. tenuifolia resulted in the reduction in the density of peltate glandular trichomes, the expression of key genes related to mono-terpene synthesis, and the relative content of limonene and pulegone, the main components of monoterpene. These results suggested that StHD1 and StHD8 of S. tenuifolia formed a complex to regulate glandular trichomes and affect the biosynthesis of monoterpenes.
Trichomes/metabolism* ; Lamiaceae/genetics* ; Tobacco/genetics* ; Monoterpenes/metabolism* ; Cloning, Molecular ; Plant Proteins/metabolism* ; Gene Expression Regulation, Plant

PURPOSE: A decline in the circulating levels of estrogen impairs vaginal engorgement, which leads to histopathological changes in the vaginal tissues. The aim of this study was to evaluate the effect of delayed estrogen replacement on the vagina structure in castrated rabbits. Materials and Methods: New Zealand White female rabbits were randomly divided into three groups; two groups (the castration and castration estrogen replacement groups) were castrated: the control group underwent sham operations. Nine weeks after surgery, the estrogen replacement group received subcutaneous injections of estrogen (50microgram/kg/day) for 4 weeks. Vaginal tissue was processed for histology with Masson's trichome stain. The expressions of e-NOS and n-NOS were examined by immunohistochemistry and Western blot. RESULTS: From the histology, marked thinning of the vaginal epithelial layers, and decreased smooth muscle content and submucosal microvasculatures were evident, but with no increase in the collagen fibers in the castrated group was noted. The vaginal tissue of the estrogen replaced animal showed similar morphological features to those in the control rabbits. From the histomorphometry, the percentage of vaginal smooth muscle in the castrated group was significantly decreased compared to those in the control and estrogen replaced groups. The expression and signal intensity of e-NOS were decreased in the vagina of the castrated animals compare to those in the control group. In the estrogen replaced group, the expression of e-NOS was normalized. CONCLUSIONS: Delayed estrogen replacement showed structural restoration of vaginal tissues. These results suggest that delayed estrogen replacement therapy could improve the sexual function in menopausal women.
Animals ; Blotting, Western ; Castration ; Collagen ; Estrogen Replacement Therapy* ; Estrogens* ; Female ; Humans ; Immunohistochemistry ; Injections, Subcutaneous ; Menopause ; Muscle, Smooth ; New Zealand ; Rabbits ; Trichomes ; Vagina

BACKGROUND: Nettles, of the plant family Urticae, cause nonimmunologic contact urticaria by stinging hairs. Light application of the leaf or stem to the skin results in the rapid development of a short-lived stinging sensation accompanied by wheal formation and itching. The types of chemical mediators causing the stinging sensation and wheal have not yet been clarified. OBJECTIVE: To investigate stinging and urticating substances causing contact urticaria when the skin is in contact with nettle(Urtica thunbergiana). METHODS: Twenty two volunteers were used for the clinical portion of this investigation. The time to wheal development and the evanescent time of pain and wheal were measured after pricking the forearm skin with nettle trichomes, following the administration of antihistamine, antiserotonin or corticosteroid, respectively or a combination of two, either with or without the application of substance P antagonist(capsaicin cream, 0.025%). Nettle extracts were prepared from dry stems and leaves of Urtica thunbergiana. Histamine, serotonin and substance P(SP) contents of nettle extracts were measured by gas chromatographic mass spectroscopy. RESULTS: After puncture of the skin by trichome of nettle, a stinging sensation developed within a few seconds but disappeared faster at the site where capsaicin cream had been applied than at the nonapplied skin site(p<0.05). The development and evanescent time of wheal were not significantly correlated with the application of capsaicin cream. The evanescent time of pain after pricking the skin with trichome following the administration of antiserotonin was significantly shortened at the nonapplied skin site. Pain disappeared more rapidly at the capsaicin applied site following the administration of antihistamine or antiserotonin than at the nonapplied sites(p<0.05), but not following prednisolone administration. Wheal development was delayed significantly at both the capsaicin applied and nonapplied sites after antihistamine administration(p<0.05). Histamine was detected only in the nettle extract. SP and serotonin were not detected because of their insolubility in solvents. CONCLUSION: These results suggest that wheal-and-flare reaction to nettle stings is due primarily to the histamine and partly to the serotonin and SP introduced by the nettle. SP may be mainly involved and partially involved by serotonin in stinging pain.
Bites and Stings ; Capsaicin ; Forearm ; Hair ; Histamine ; Humans ; Mass Spectrometry ; Plants ; Prednisolone ; Pruritus ; Punctures ; Sensation ; Serotonin ; Skin ; Solvents ; Substance P ; Trichomes ; Urticaria* ; Volunteers

This study was designed to clarify morphological changes in the renal corpuscles of the experimental rabbit kidney after unilateral ureteral obstruction and release of Unilateral ureteral obstruction through a light and electro microscopy. It was further aimed at obtaining data, esp. of electron microscopy, concerning the morphological repair after relieving ureteral obstruction. A total of 25 white rabbits, weighing 2.5kg, were used. 25 rabbits were divided into three groups-normal control, unilateral ureteral obstruction, release of unilateral obstruction. The normal! control group consisted of 5 rabbits. The remaining 20 rabbits were completely ligated in the left ureter with silk threads. Ten obstructed kidneys were studied by light and electron microscopy, five of them two weeks after obstruction, and the remaining five four weeks after obstruction. A total of ten postobstructed kidneys, five of which were reoperated by Mackinnon method for the relief of ureteral obstruction two weeks after obstruction and the remaining five four weeks after obstruction. They were studied for light and electron microscopy 15 days after relieving ureteral obstruction. Specimens of renal cortex from experimental animals were fixed in 10% neutral formalin or Bouin's solution, embedded in paraffin wax, sectioned at a thickness of 5 um, and stained with hematoxylin-eosin, periodic acid-schiff reaction or Masson's trichome for light microscopy. For electron microscopy, the tissues were fixed in a mixture of 2% paraformaldehyde and 2.5ao glutaraldehyde (phosphate buffer, pH 7.2) prior to fixation in 1% osmium tetroxide (phosphate buffer, pH 7.2) and then embedded in Epon 812. The sections were cut with LKB- III ultramicrotome and contrasted with uranyl acetate and lead citrate and examined with electron microscopy JEM 100B. The results of this study were as follows: 1. The abnormal morphology of the glomerulus in kidneys obstructed for four weeks showed: Slight glomerular congestion, capillary dilatation, no changes in endothelium, podocyte and microstructures. 2. Deformity of Bowman's capsular epithelium such as irregular basal invaginations facing with the basal lamina was observed in the two week obstructed kidneys, and severe degenerative changes such as desquamated capsular epithelial cells in the Bowman's space and long basal cytoplasmic processes embedded in the basal lamina in the four week obstructed kidneys. 3. Among the postobstructed kidney, the two week obstructed kidneys were restored to near normal on the 15th day, but in the four week obstructed kidney, Bowman's capsular epithelium showed partial recovery. 4. The results indicate that glomeruli in the renal corpuscle were preserved until four week weeks after ureteral obstruction but Bowman's epithelium showed change in the 2nd week after obstruction and severe degenerative changes of Bowman's capsular epithelium were noticed in the 4th week after ureteral obstruction. The morphological changes were totally restored in the group where ureteral obstruction was relieved after two weeks, but in the group which was relieved of ureteral obstruction after four weeks the morphological repairs were prolonged until after 15 days.
Animals ; Basement Membrane ; Capillaries ; Citric Acid ; Congenital Abnormalities ; Cytoplasm ; Dilatation ; Endothelium ; Epithelial Cells ; Epithelium ; Estrogens, Conjugated (USP) ; Formaldehyde ; Glutaral ; Hydrogen-Ion Concentration ; Kidney ; Microscopy ; Microscopy, Electron ; Osmium Tetroxide ; Paraffin ; Periodic Acid-Schiff Reaction ; Podocytes ; Rabbits ; Silk ; Trichomes ; Ureter* ; Ureteral Obstruction*