Malus Sylvestris Research Articles

The vulnerability of US apple (Malus) genetic resources

Apple (Malus × domestica Borkh.) is one of the top three US fruit crops in production and value. Apple production has high costs for land, labor and inputs, and orchards are a long-term commitment. Production is dominated by only a few apple scion and rootstock cultivars, which increases its susceptibility to dynamic external threats. Apple crop wild relatives, including progenitor species Malus sieversii (Ledeb.) M. Roem., Malus orientalis Uglitzk., Malus sylvestris (L.) Mill., and Malus prunifolia (Willd.) Borkh., as well as many other readily hybridized species, have a wide range of biotic and abiotic stress resistances as well as desirable productivity and fruit quality attributes. However, access to wild materials is limited and wild Malus throughout the world is at risk of loss due to human encroachment and changing climatic patterns. The USDA-ARS National Plant Germplasm System (NPGS) Malus collection, maintained by the Plant Genetic Resources Unit in Geneva, NY, US is among the largest collections of cultivated apple and Malus species in the world. The collection currently has 5004 unique accessions in the field and 1603 seed accessions representing M. × domestica, 33 Malus species, and 15 hybrid species. Of the trees in the field, 3,070 are grafted and are represented by a core collection of 258 individuals. Many wild species accessions are represented as single seedlings (non-grafted). The crop vulnerability status of apple in the US is moderate because although there are a few breeders developing new commercial cultivars who also access Malus species, threats and challenges include new diseases, pests, and changing climate combined with industry needs and consumer demands, with a limited number of cultivars in production.

Effect of Treatment Time of 1-MCP on Ripening of ‘Fuji’ Apples Stored at Low and Room Temperature for a Long Period

The effect of 1-methycyclopropene (1-MCP) treatment at 0, 2, and 4 d after harvest (0 DAH, 2 DAH, and 4 DAH) of apples [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] was evaluated on several fruit quality parameters for 180-d at 0.5°C in the air, followed by 28-d at room temperature. The 1,000 nl l-1 1-MCP treatments were adequate to maintain fruit titratable acidity (TA) and firmness, inhibiting the rates of ethylene production and respiration during the storage of both cold and the room temperatures, regardless of the 1-MCP treatment time.

Species complexes of predatory mites and spider mites (Acari: Phytoseiidae, Tetranychidae) on cultivated and wild apple trees in Serbia

The species complexes of leaf-inhabiting mites on apples were studied on actively cultivated and neglected apple orchards as well as on wild apple trees in Serbia, from 1981 to 2013. Thirty-nine species of 11 families were found on Malus domestica Borkh; 25 species of 6 families were found on Malus sylvestris (L.) Miller and Malus pumila Miller. A complex of 18 phytoseiid and 9 spider mite species was detected. Four phytoseiid and three spider mite species were found in commercial apple orchards, 13 phytoseiid and 5 spider mite species in neglected apple stands, and 12 phytoseiid and 7 spider mite species in forest habitats. Predatory mites Euseius finlandicus (Oudemans) and Amblyseius andersoni (Chant) and spider mites Panonychus ulmi Koch and Amphitetranychus viennensis (Zacher) had the highest constancy of occurrence. Positive and significant (or very significant) association was detected for 16 pairs of species. The pairs of phytoseids-spider mites Amb. andersoni–Pan. ulmi, Amb. andersoni–Tetranychus turkestani Ugarov et Nikolskii, and Typhlodromus (Anthoseius) rhenanus (Oudemans)–Te. turkestani had the highest interspecific association indices. Higher mite diversity was found on wild than on cultivated apples, and higher diversity on neglected than on cultivated apples. The complexes of phytoseiids and spider mites had uniform composition in the cultivated orchards, while their composition in the neglected stands was nonuniform and extremely heterogeneous on the wild apple trees.

Inferring the origin of rare fruit distillates from compositional data using multivariate statistical analyses and the identification of new flavour constituents.

In Serbia, delicatessen fruit alcoholic drinks are produced from autochthonous fruit-bearing species such as cornelian cherry, blackberry, elderberry, wild strawberry, European wild apple, European blueberry and blackthorn fruits. There are no chemical data on many of these and herein we analysed volatile minor constituents of these rare fruit distillates. Our second goal was to determine possible chemical markers of these distillates through a statistical/multivariate treatment of the herein obtained and previously reported data. Detailed chemical analyses revealed a complex volatile profile of all studied fruit distillates with 371 identified compounds. A number of constituents were recognised as marker compounds for a particular distillate. Moreover, 33 of them represent newly detected flavour constituents in alcoholic beverages or, in general, in foodstuffs. With the aid of multivariate analyses, these volatile profiles were successfully exploited to infer the origin of raw materials used in the production of these spirits. It was also shown that all fruit distillates possessed weak antimicrobial properties. It seems that the aroma of these highly esteemed wild-fruit spirits depends on the subtle balance of various minor volatile compounds, whereby some of them are specific to a certain type of fruit distillate and enable their mutual distinction.

Effect of Some Growth Regulators on the Growth of Hendersonula Toruloidea Nattrass

Effect of different concentrations of indole-3 ylacetic acid, indole butyric acid and alpha-naphthalein acetic acid on the growth of Hendersonula toruloidea Nattrass, causing brown rot of fruits of Malus sylvestris Mill was studied. It was observed that indole acetic acid and alpha-naphthalein acetic acid supported good growth of the fungus in lower concentrations (0.1 & 0.5 ppm) but at 50 ppm the growth was inhibited. Indole butyric acid promoted the growth at the concentration of 50 ppm but an inhabitation in growth was observed at lower concentrations.

Internal ethylene concentrations in apple fruit at harvest affect persistence of inhibition of ethylene production after 1-methylcyclopropene treatment

Factors that affect the efficacy of 1-methycyclopropene (1-MCP) treatment of apples [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] include cultivar and maturity. In this study, ‘McIntosh’, ‘Cortland’ and ‘Empire’ apples were categorized by internal ethylene concentrations (IECs) at harvest, treated with 1μLL−1 1-MCP, and the IECs of individual fruit followed at 30d intervals during air storage at 0.5°C for 90d. IECs at harvest ranged from <0.5μLL−1 to ≥100μLL−1, 51<100μLL−1, and 10<50μLL−1 for ‘McIntosh’, ‘Cortland’ and ‘Empire’, respectively. 1-MCP treatment resulted in a decrease of IECs in fruit of all cultivars by day 30 after harvest. During subsequent storage IECs remained low in fruit with <1μLL−1 at harvest, but in ‘McIntosh’, ‘Cortland’ increased in proportion to IECs at harvest, but not in ‘Empire’. The importance of initial IECs in fruit on the persistence of 1-MCP inhibition of ethylene production was confirmed in a further experiment, in which IECs in untreated and 1-MCP treated ‘McIntosh’ and ‘Empire’ apples were measured for up to 194d. 1-MCP also decreased 1-aminocyclopropene-1-carboxylic acid (ACC) concentrations in fruit. The results of our study are consistent with the hypothesis that IEC modulates the sensitivity of climacteric fruit to 1-MCP.

Wild European apple (Malus sylvestris (L.) Mill.) population dynamics: insight from genetics and ecology in the Rhine Valley. Priorities for a future conservation programme.

The increasing fragmentation of forest habitats and the omnipresence of cultivars potentially threaten the genetic integrity of the European wild apple (Malus sylvestris (L.) Mill). However, the conservation status of this species remains unclear in Europe, other than in Belgium and the Czech Republic, where it has been declared an endangered species. The population density of M. sylvestris is higher in the forests of the upper Rhine Valley (France) than in most European forests, with an unbalanced age-structure, an overrepresentation of adults and a tendency to clump. We characterize here the ecology, age-structure and genetic diversity of wild apple populations in the Rhine Valley. We use these data to highlight links to the history of this species and to propose guidelines for future conservation strategies. In total, 255 individual wild apple trees from six forest stands (five floodplain forests and one forest growing in drier conditions) were analysed in the field, collected and genotyped on the basis of data for 15 microsatellite markers. Genetic analyses showed no escaped cultivars and few hybrids with the cultivated apple. Excluding the hybrids, the genetically “pure” populations displayed high levels of genetic diversity and a weak population structure. Age-structure and ecology studies of wild apple populations identified four categories that were not randomly distributed across the forests, reflecting the history of the Rhine forest over the last century. The Rhine wild apple populations, with their ecological strategies, high genetic diversity, and weak traces of crop-to-wild gene flow associated with the history of these floodplain forests, constitute candidate populations for inclusion in future conservation programmes for European wild apple.

Species distribution, host affinity and genetic variability of Heterobasidion annosum sensu lato in the Czech Republic

SummaryThe distribution and host affinity of Heterobasidon annosum (Fr.) Bref. sensu lato (Basidiomycota, Bondarzewiaceae) in the Czech Republic was investigated. Sampling was undertaken in natural forests, in stands with former presence of fir and spruce and in forests on formerly agricultural lands. The identification and phylogenetic relationship of the species complex was studied by comparing DNA sequences of three gene regions: glyceraldehyde 3‐phosphate dehydrogenase (G3P), translation elongation factor 1‐alfa (EFA) and transcription factor (TF). The species relationships were demonstrated by haplotype network and Bayesian phylogram construction for the combined data set. The distribution of each species was demonstrated. Heterobasidion annosum s.s., which had the greatest host range, was found on thirteen genera (Pinus, Picea, Fraxinus, Betula, Corylus, Alnus, Abies, Acer, Salix, Ligustrum, Quercus, Larix and Prunus). Heterobasidion parviporum was found on spruce (Picea abies), fir (Abies alba), apple tree (Malus sylvestris), Vitis vinifera and Fagus sylvatica, and Heterobasidion abietinum was observed on fir (Abies), spruce (Picea), pine (Pinus) and maize (Zea mays). Sequences from different genes yielded conflicting results for six specimens, which were interpreted as belonging to interspecific hybrids. These were collected from A. alba, P. abies and Chamaecyparis lawsoniana. The occurrence of Heterobasidion annosum s.l. is strongly associated with the natural distribution of its hosts and the occurrence of H. abietinum appears to be related to the historical occurrence of A. alba in stands now reforested with P. abies. No statistically significant differences in the vertical distributions within the species complex were detected.

Diet of the Italian hare (Lepus corsicanus) in a semi-natural landscape of southern Italy

The food habits of the endangered Italian hare have not received adequate attention from researchers. In this study, the diet composition of this species and its sea- sonal variation were assessed by analysing faecal pellets in a semi-natural landscape in the south of Italy. The results showed that hares feed on 62 species of plants during the year, with a conspicuous presence of herbaceous ones (e.g., Trifolium pratense , Brachypodium sylvaticum , Festuca arun- dinacea ) as these occurred at high frequencies in most of the faecal samples. In spring, diet composition was charac- terised by a high percentage of Graminaceae ( > 37%). In the other seasons, hares also included fruits (e.g., Prunus spi- nosa , Pyrus piraster , Malus sylvestris ), which, in autumn, accounted for > 27%. There were significant differences among seasons (p 70%) in the diets during the other seasons with a more pronounced similarity between summer and autumn (S o rensen, C s = 0.80; Morisita-Horn, C MH = 0.73).

News of the Société Botanique de France

Despite its economic, cultural and historical importance, few studies have investigated the evolutionary history of the domesticated apple (Malus domestica) as well as those of its wild relatives. Using new population genetic approaches (approximate Bayesian computation) with microsatellites and nuclear sequences, this thesis aimed to unravel, at different evolutionary scales (phylogeography, speciation, domestication), the natural and artificial diversification processes at play in the Malus genus. Research focused on the four wild apple species distributed across Eurasia [Malus orientalis (Caucasus), Malus sieversii (Central Asia), Malus sylvestris (Europe) and Malus baccata (Siberia)] and on the single domesticated apple species in the genus, Malus domestica. The thesis was divided into four parts: (1) domestication history of the cultivated apple, from its origin in Central Asia to Europe, (2) post-glacial recolonization history of the European crabapple (M. sylvestris), (3) the history of speciation among the five Malus species, (4) crop-to-wild gene flow and dispersal capacities of the closest wild relative species (M. sylvestris, M. sieversii and M. orientalis). By investigating artificial diversification, we found evidence of unique processes of domestication in this fruit tree, with no bottleneck and with extensive post-domestication introgressions by a wild species (M. sylvestris) other than the ancestral progenitor (M. sieversii). Natural diversification patterns (phylogeography, speciation and population structure) revealed large effective population sizes, high dispersal capacities and weak spatial genetic structures. This thesis also revealed high levels of interspecific hybridizations, and a particularly high level of crop-to-wild gene flow in Europe and Central Asia. This study extended our knowledge of population structures for the wild species that contributed to the cultivated apple genome, as well as the extent of hybridization rates. This work is essential for the conservation of wild apple populations, the maintenance of the integrity of wild species facing fragmentation and future breeding programmes concerning the domesticated apple.

Morphology and stucture of wild apple (Malus silvestris Mill.).common pear (Pyrus cofnmunis L.) and Chaenomeles japonica (Thunb) Lindl. seeds

The outer and inner structure of wild apple (&lt;em&gt;Malus silvestris&lt;/em&gt; Mill.), common pear (&lt;em&gt;Pyrus communis&lt;/em&gt; L.) and &lt;em&gt;Chaenomeles japonica&lt;/em&gt; (Thunb.) Lindl. seeds was investigated. It was found that the outer structure exhibits good diagnostic features expressed in the first place in the relief of the seed coat and further in the arrangement and appearance of the site of attachment of the free end of the funiculus and the shape of the seeds. In ripe seeds there is, under the thick seed coat, an endosperm layer completely surrounding the embryo which has large cotyledons and a thick rootlet.

Apples as enantiospecific bioreagents in the hydrolysis of racemic esters and oxygenation of alcohols obtained during the process

Biotransformation carried out by apples (&lt;em&gt;Malus silvestris&lt;/em&gt;) resulted in enantiospecific hydrolysis of racemic acetates, the derivatives of secondary aromatic-aliphatic or terpenic alcohols. The alcohols are oxygenated to ketones (reversible reaction). The differences in the rate of the hydrolysis of enantiomeric esters enable isolation of pure, unreacted R-acetates.

The ability of fruit and vegetable enzyme system to hydrolyse ester bonds

The pulp of potato tubers (&lt;i&gt;Solanum tuberosum&lt;/i&gt;), topinambur (&lt;i&gt;Helianthus tuberosus&lt;/i&gt;) and apples (&lt;i&gt;Malus silvestris&lt;/i&gt;) can hydrolyse totally, or almost totally, ester bonds in phenyl, α- and β-naphthyl, benzyl and cinnamyl acetates. In methyl 4-acetoxy-3-metoxybenzoate and methyl 2,5-diacetoxybenzoate as well as testosterone propionate and 16,17-acetonide of 21-acetoxy-6-fluoro-16α,17β,21-trihydroxy-4-pregnen-3,20-dione, the hydrolysis is selective towards the substrate and the bioreagent. In contrast, ethyl benzoate and cinnamate are resistant to hydrolysis.

Fruit Size Affects Physiological Attributes and Storage Disorders in Cold-stored ‘Royal Gala’ Apples

‘Royal Gala’ apple [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] fruit can be susceptible to the development of postharvest disorders such as flesh breakdown and cracking (splitting) during and after cold storage. The objective of this research was to investigate fruit size and 1-methylcyclopropene (1-MCP) treatment effects on fruit physiological attributes and incidence and severity of storage disorders in ‘Royal Gala’ apples held in cold storage. In 2011, fruit segregated at harvest into two groups based on size (120 to 175, 250 to 350 g/fruit) were stored in air at 0.5 °C for 6 months and then at 20 °C for 7 days. In 2012, fruit were sorted into four groups (less than 200, 200 to 240, 241 to 280, and greater than 280 g/fruit), treated with 0 or 1 μL·L−1 1-MCP for 12 hours, and then stored in air at 0.5 °C for 3 or 6 months. Storage disorders were only detected at 6 months, regardless of 1-MCP treatment. In both control and 1-MCP-treated fruit, flesh breakdown incidence increased with fruit size, whereas severity was less associated with size. The progression of flesh breakdown developed in overall cortex tissue of control fruit but only detected in the stem-end tissue of 1-MCP-treated fruit. Internal ethylene concentration (IEC) decreased and CO2 production increased with increased fruit weight; however, 1-MCP-treated fruit had low IEC regardless of weight. Cortex tissue lightness (L*) increased with fruit size irrespective of tissue localization (stem end, equatorial, calyx end) at harvest. During 6 months’ storage, L* decreased with increased fruit size in controls but not 1-MCP-treated fruit. Fruit fresh weight loss increased with fruit size and storage duration, more so in controls when compared with 1-MCP-treated fruit. Furthermore, fruit circumference increased during storage with fruit size only for control fruit. These physical changes are associated with susceptibility of large fruit to flesh breakdown more so than small fruit. Reduced flesh breakdown incidence, progression of symptoms from the stem end into the cortex, and symptom severity in 1-MCP-treated fruit may indicate flesh breakdown is related to fruit ripening and senescence.

Exogenous dormancy-breaking substances positively change endogenous phytohormones and amino acids during dormancy release in ‘Anna’ apple trees

A 2-season trial was conducted to verify the effects of foliar applications of some dormancy-breaking substances (DBS) on dormancy release in buds of ‘Anna’ apple (Malus sylvestris, Mill) trees, as well as on metabolic changes in the contents of phytohormones, proline and arginine in buds during their release from dormancy. The efficiency of early bud break induced by Dormex™, potassium nitrate, mineral oil, calcium nitrate and thiourea was noticed in varying degrees. Although Dormex™ was distinguished, all DBS hastened bud break, shortened flowering duration, improved bud break% and fruit-set%, increased the contents of gibberellic acid, indole-3-acetic acid, proline and arginine, but reduced abscisic acid content in buds as compared to the control. These results were positively reflected in the final tree yield. Accordingly, it is concluded that the use of Dormex™, at a rate of 4 %, could be recommended for reaching bud break as early as possible and improving ‘Anna’ apple tree yield under the short winters in Egypt and similar regions by regulating the contents of proline, arginine and phytohormones in buds.

Phylogenetic Analysis of 47 Chloroplast Genomes Clarifies the Contribution of Wild Species to the Domesticated Apple Maternal Line

Both the origin of domesticated apple and the overall phylogeny of the genus Malus are still not completely resolved. Having this as a target, we built a 134,553-position-long alignment including two previously published chloroplast DNAs (cpDNAs) and 45 de novo sequenced, fully colinear chloroplast genomes from cultivated apple varieties and wild apple species. The data produced are free from compositional heterogeneity and from substitutional saturation, which can adversely affect phylogeny reconstruction. Phylogenetic analyses based on this alignment recovered a branch, having the maximum bootstrap support, subtending a large group of the cultivated apple sorts together with all analyzed European wild apple (Malus sylvestris) accessions. One apple cultivar was embedded in a monophylum comprising wild M. sieversii accessions and other Asian apple species. The data demonstrate that M. sylvestris has contributed chloroplast genome to a substantial fraction of domesticated apple varieties, supporting the conclusion that different wild species should have contributed the organelle and nuclear genomes to the domesticated apple.

Fruit maturity, controlled atmosphere delays and storage temperature affect fruit quality and incidence of storage disorders of ‘Fuji’ apples

The effects of storage temperature, harvest maturity, controlled atmosphere (CA) storage regime, and CA strategies on fruit quality and the incidence of storage disorders during storage of ‘Fuji’ apples [Malus sylvestris (L.) Mill var. domestica (Borkh.) Mansf.] were investigated. Flesh browning incidence was lower in fruit stored at 2°C than at 0°C, but fruit were also softer and had lower titratable acidity (TA) at the higher storage temperature. All subsequent studies were carried out at 0°C. High partial pressures of CO2 (pCO2) of 2.5kPa compared with <1kPa did not consistently affect flesh firmness, soluble solids concentration (SSC), and TA. The incidence of flesh browning was higher in more mature fruit and it was further aggravated by higher pCO2 in the storage atmosphere, irrespective of pO2, while watercore disappearance was more rapid at <1kPa CO2 than at 2.5kPa CO2. Flesh browning was also higher at 1.5kPa CO2 than at <1kPa CO2, although ethylene production was lowest at 1.5kPa pCO2. Stepwise delay of high CO2 or low O2 CA storage did not affect flesh firmness, TA, internal ethylene concentration, respiration rate, and ethylene production, but watercore disappearance was more rapid, compared with standard CA storage at 2 months. Flesh browning was not detected in the stepwise CO2 delayed CA treatment and low CO2 CA storage (<0.2kPa CO2), but a low (<4%) incidence of flesh browning was found in a delayed CA treatment and standard CA storage. These results indicate that ‘Fuji’ apples could be stored at 0°C in 1–2kPa O2/<1kPa CO2 if harvested at less than 190 days after full bloom. Stepwise CO2 delay CA or low CO2 CA storage regimes are recommended to retard the loss of fruit quality factors and reduce development of CA-related storage disorders.

Postglacial recolonization history of the European crabapple (Malus sylvestris Mill.), a wild contributor to the domesticated apple

Understanding the way in which the climatic oscillations of the Quaternary Period have shaped the distribution and genetic structure of extant tree species provides insight into the processes driving species diversification, distribution and survival. Deciphering the genetic consequences of past climatic change is also critical for the conservation and sustainable management of forest and tree genetic resources, a timely endeavour as the Earth heads into a period of fast climate change. We used a combination of genetic data and ecological niche models to investigate the historical patterns of biogeographic range expansion of a wild fruit tree, the European crabapple (Malus sylvestris), a wild contributor to the domesticated apple. Both climatic predictions for the last glacial maximum and analyses of microsatellite variation indicated that M. sylvestris experienced range contraction and fragmentation. Bayesian clustering analyses revealed a clear pattern of genetic structure, with one genetic cluster spanning a large area in Western Europe and two other genetic clusters with a more limited distribution range in Eastern Europe, one around the Carpathian Mountains and the other restricted to the Balkan Peninsula. Approximate Bayesian computation appeared to be a powerful technique for inferring the history of these clusters, supporting a scenario of simultaneous differentiation of three separate glacial refugia. Admixture between these three populations was found in their suture zones. A weak isolation by distance pattern was detected within each population, indicating a high extent of historical gene flow for the European crabapple.

Comparison of the nectary structure of chosen species from subf. Pomoideae (Rosaceae)

The research of the anatomy of nectaries of six species from &lt;em&gt;Pomoideae&lt;/em&gt; was carried out. The observations concerned the flowers longitudinal sections of the following species: &lt;em&gt;Malus silvestris&lt;/em&gt; Mill., &lt;em&gt;Pyrus communis&lt;/em&gt; L., &lt;em&gt;P. ussuriensis&lt;/em&gt; Maxim., &lt;em&gt;Cydonia oblonga&lt;/em&gt; Mill., &lt;em&gt;Chaenomeles japonica&lt;/em&gt; Ldl. and &lt;em&gt;Ch. speciosa&lt;/em&gt; Nakai. It was stated, that the nectary glands distinguished themselves by the dimension of section surface, thickness of layers, the sizes of glandular and epidermis cells. A considerable difference of external wall thickness among epidermis cells was also observed. The nectaries of the smallest size, consisting of the tiniest cells occurred in &lt;em&gt;Malus silvestris&lt;/em&gt;. Whereas, the biggest ones were possessed by &lt;em&gt;Cydonia oblonga&lt;/em&gt;. Here the nectaries had maximum section surface, thickness, number of layers and cell dimension. The nectary epidermis of &lt;em&gt;Pyrus&lt;/em&gt; was characterized by the thinnest external wall. The Chaenomeles speciosu and &lt;em&gt;Cydonia oblonga&lt;/em&gt;, by the thickest one with a massive cuticle layer. The vascular bundles reaching the nectary tissue consisted of phloem and xylem. The positive dependence between the nectary dimension and the abundance of nectar secretion in some species, and the occurence of the opposite tendency in the others is discussed.

Pharmacognostic and high performance thin layer chromatography finger printing of Pyrus malus Linn

Background: Pyrus malus L. Syn. Malus sylvestris Mill., commonly known as 'Apple' is a well known tree belonging to the family Rosaceae. Leaves and young aerial parts are used as medicine in homoeopathy. Objective: The Pharmacognostic and physico-chemical studies are carried out to facilitate use of correct species and standard raw materials. Material and Methods: Pharmacognostic examinations of aerial parts (leaf, petiole and stem) of authentic sample of Pyrus malus L., have been carried out. Physico-chemical parameters of raw drug viz., extractive value, ash values, formulation besides weight per mL, total solids, alcohol content along with high performance thin layer chromatography (HPTLC) and UV-visible studies have been worked out for mother tincture. Results: The leaves are ovate to oblong, margin serrate, acute, glabrous above and tomentose beneath with a short petiole. The leaf is hypostomatic with anomocytic stomata. Uniseriate macroform flagellate conical and unicellular filiform cylindrical hairs occur on the leaf. In transverse section (TS) the midvein is conspicuously ridged on abaxial side. Epidermis is 1-layered with larger cells on abaxial. Cells over midvein are papillated. Palisade is 2-layered. The vascular bundle at midvein is single, large and arc shaped. The petiole in T.S. is shield like with an adaxial groove. The cortex is aerenchymatous towards adaxial. The vascular tissue consists of a single, large, arcuate bundle and two small adaxial bundles. The young stem is oval to spherical. The vascular tissue is in the form of a continuous cylinder. 1-3 seriate rays are present in the xylem. The xylem towards the centre possess secretory canals arranged in a ring. Conclusion: The powder microscopic features and organoleptic characters along with the anatomical and physico-chemical studies are diagnostic to establish the standards for the drug.