Reddish Brown Research Articles

Synthesis of silver nanoparticles assisted by aqueous root and leaf extracts of Rhus chinensis Mill and its antibacterial activity

In the present study, silver nanoparticles were synthesized using aqueous root and leaf extracts of Rhus chinensis Mill. This study aimed to undertake the green synthesis of silver nanoparticles utilizing plant extracts in an eco-friendly, cost-effective, and more efficient manner with its antibacterial application. The prepared silver nanoparticles (AgNPs) were characterized by using different techniques. Such as ultraviolet–visible spectroscopy (UV–Vis), Fourier transform infrared spectroscopy (FT-IR), x-ray diffraction spectroscopy (XRD), field emission-scanning electron microscopy (FE-SEM), and energy dispersive X-ray (EDX). The color changes from yellowish to reddish brown can be visualized and it indicates the formation of silver nanoparticles. The UV–Vis absorption peak shown by the synthesized AgNPs assisted by root and leaf extract was at 443 nm and 440 nm respectively. The functional group present in plants' secondary metabolites may act as capping and stabilizing agents, indicated by the shifting and disappearing of the peak in the plant extracts and the extracts-assisted synthetic nanoparticles. The crystallite size of synthesized AgNPs assisted by the root and leaf extracts of Rhus cinensis was found to be 11.01 nm and 13.39 nm respectively, while with the help of FE-SEM image the shape and particle size of synthesized AgNPs root and leaf extract was found spherical with particle diameter of 54.40 nm and 30.89 nm respectively. The presence of an intense silver component was confirmed by EDX analysis which showed an intense peak at around 3 Kev and other elements like Cl, O, C, and N were also reported in synthesized AgNPs. Both the plant extracts assisted synthesized AgNPs showed higher zones of inhibition (ZOI) against both the Gram-positive and Gram-negative bacteria. The results of the study indicate the potential benefit of synthesized silver nanoparticles using Rhus chinensis root and leaf extracts for biomedical purposes.

A Comparative Analysis on the Material Properties of Paintings in Modern Buildings: Focusing on Seoul Weather Station

In this study, scientific analysis(cross-section and component analysis) was conducted by selecting paint samples painted on the window frames and walls of the Seoul Weather Station. The color of the window frame is reddish brown and green. The reddish brown is mainly red iron oxide(Fe<sub>2</sub>O<sub>3</sub>), mixed with titanium white(TiO<sub>2</sub>), and in some layers, red lead oxide(Pb<sub>3</sub>O<sub>4</sub>) or lead(II) chromate(PbCrO<sub>4</sub>) is identified, and the green pigment is considered organic pigments. The resin used in the window frame is Alkyd Resin. The color of the wall is divided into pale blue green, white, and ivory colors, and the white color of the wall is calcium carbonate(CaCO<sub>3</sub>) and titanium white(TiO<sub>2</sub>), and the pale blue green is the phthalocyanine green. It is determined that paint containing acrylic resin was used for the inner wall. The characteristics of paint were compared and considered with the walls of the Seoul Weather Station, the columns of the Grand Greenhouse of Changgyeonggung Palace, and the outer wall of Busan Songjeong Station. White paint, a common color identified in the three buildings, is identified as two types of paint mixed with zinc White(ZnO) and calcium carbonate(CaCO<sub>3</sub>), and titanium white(TiO<sub>2</sub>) and calcium carbonate(CaCO<sub>3</sub>). It seems that the orange paint uses red lead oxide(Pb<sub>3</sub>O<sub>4</sub>) or Lead(II) chromate(PbCrO<sub>4</sub>) and the green paint uses organic pigments. Based on the above research results, paint painted on buildings is believed to have been painted several times after the building was built, and natural science research should be carried out to determine the history.

Characterizing the Size Distribution of Silver Nanoparticles Biofabricated Using Calotropis gigantea from Geothermal Zone

This research aims to synthesize silver nanoparticles (AgNPs) using an aqueous leaf extract of Calotropis gigantea obtained from the geothermal manifestation Ie Seu-Um, Aceh Besar, Aceh Province, Indonesia. The C. gigantea leaf extract was mixed with AgNO3 solutions at concentrations of 2, 5, and 9 mM, respectively. The mixture was stirred at 80 rpm by a magnetic stirrer for 48 hours in the dark. The change in solution color indicated the reduction of Ag+ to Ag0. The resulting AgNPs synthesized using C. gigantea leaf extract (AgNPs-LCg) exhibited cloudy grey, reddish dark brown, and light brown colors when synthesized with AgNO3 concentrations of 2, 5, and 9 mM, respectively. The particle sizes of AgNPs-LCg had maximum frequencies at 246.98 nm (synthesized using AgNO3 2 mM), 93.02 nm (synthesized using AgNO3 5 mM), and 171.25 nm (synthesized using AgNO3 9 mM). The zeta potential values of AgNPs-LCg using 2, 5, and 9 mM AgNO3 were -41.9, -40.1, and -31.4 mV, respectively. Based on the solution color, nanoparticle size, and stability value of AgNPs, it can be concluded that the use of AgNO3 at 5 mM is optimal for the green synthesis process of AgNPs-LCg.

Anti-angiogenic activity of Ficus deltoidea L. Jack silver nanoparticles using chorioallantoic membrane assay

Background: Ficus deltoidea L. Jack is a folk medicinal plant known for its pharmacological properties, including anti-inflammatory, anticancer and anti-angiogenic. This study aimed to evaluate the anti-angiogenic properties of silver nanoparticles biosynthesized using F. deltoidea leaf extract (AgNPs-Fd). Methods: The AgNPs-Fd were synthesized by mixing 100 mL 1 mM aqueous silver nitrate (AgNO3) and 100 mL 0.1% F. deltoidea ethanolic leaf extract. The resulting AgNPs-Fd were observed for color change and Tyndall effects. Reaction mixture color change from pale brown to reddish brown was observed at 48 h at 37°C. The characterization of AgNPs-Fd was completed with UV–Vis spectroscopy, transmission electron microscopy (TEM), X-ray diffraction (XRD) and Fourier transform infrared (FTIR) spectroscopy. For quantitative analysis of the vascular network in the chorioallantoic membrane (CAM) assay, AngioTool open-source software was used. Results: The plasmon resonance peak for AgNPs-Fd at 430 nm was visible in the UV-Visible spectrum, indicating the formation of AgNPs-Fd. The F. deltoidea extract and nanoparticles interacted well according to FTIR analysis. The AgNPs-Fd morphology of 20 nm particle sizes was observed using TEM. The chromatographic analysis of AgNPs-Fd identified potential anti-angiogenic compounds, such as phytol, stigmasterol, lupeol and sitosterol. The angiogenic inhibition properties of AgNPs-Fd were tested using the CAM assay. The 90 mg dose AgNPs-Fd treatment in CAMs demonstrated significant anti-angiogenesis, indicating effectiveness in controlling vessel formation. Conclusions: The present study suggests that eco-friendly work and the “green” process of AgNPs-Fd is potentially applicable for nanobiotechnology in anti-angiogenic fields.

Green synthesis of silver nanoparticles with an antibacterial activity using Salvia officinalis aqueous extract

This report searched the rapid and simple green biosynthesis of silver nanoparticles (AgNPs) using Salvia officinalis aqueous extract. The visual color change from colorless/yellow to reddish brown was noticed indicating the phytosynthesis AgNPs during the process. The presence of functional groups in plant extract acted as reducing and capping agents for the formation of AgNPs was confirmed by FTIR. The fabricated AgNPs were characterized by ultraviolet–visible spectroscopy, X-ray diffraction (XRD), scanning electron microscopy (SEM), and energy-dispersive X-ray analysis (EDX). The UV–Visible spectrum showed a characteristic absorption peak for Ag at around 457 nm with optical bandgaps of about 2.22 eV of AgNPs. XRD patterns at (111), (020), (022), (131), and (222) lattice planes depicted the production of face-centered cubic silver nano-structure with approximate crystal size ranges from≈20 nm by Scherrer’s formula. The predominantly spherical nanoparticles having an average size of 20 nm showed enhanced antibacterial activity against Salmonella typhimurium, Pseudomonas aeruginosa, Staphylococcus aureus, and Escherichia coli with more activity toward P. aeruginosa. Our green synthesis protocol required only 15 min for the conversion of silver ions into silver nanoparticles by heating is an efficient, rapid, simple, sustainable, cheap, and energy-efficient, alternative to conventional chemical/physical methods, a one-step application.

Mapping of a soybean rust resistance in PI 594756 at the Rpp1 locus.

The online version contains supplementary material available at 10.1007/s11032-023-01358-4.

Psidium guajava leaf extract mediated green synthesis of silver nanoparticles and its application in antibacterial coatings.

In this study, Psidium guajava (P. guajava) leaf extract-assisted silver nanoparticles (AgNPs) were synthesized and their antibacterial activities were investigated. The synthesized green AgNPs were characterized by various analytical techniques including UV-Vis spectroscopy, Fourier transform infrared spectroscopy (FTIR), X-ray diffractometry (XRD), field emission scanning electron microscopy (FE-SEM), energy dispersive X-ray (EDX) spectroscopy, etc. From the UV-Vis spectroscopic analysis, the formation of nanoparticles has been confirmed by the color change from light yellow to reddish brown of the solution due to the excitation of the surface plasmon resonance peak at 430 nm. In addition, the FTIR study showed the reduction of Ag ions owing to the presence of biomolecules in the leaf extract, which acted as reducing as well as capping agents. Furthermore, XRD analysis reveals the identified 2θ peaks of AgNPs at ∼39° with cubic structure. The FE-SEM micrograph illustrated the material was formed in nano-dimensions, with an average particle size of ∼12 nm and almost spherical in shape. Moreover, P. guajava-mediated AgNPs demonstrated good antibacterial activity against both Gram-positive (S. aureus) and Gram-negative (E. coli) bacterial strains. The synthesis was performed by a bio-reduction process where a bioactive agent is responsible for reducing metallic ions to metallic nanoparticles as an eco-friendly, cost-effective, non-toxic, one-step, and sustainable method. Therefore, this study may create an imperative synthetic route for the fabrication of green-AgNPs and their application in antibacterial coatings in cotton textiles.

Effect of Soil Moisture on Ammonia Volatilization from Urea Applied Alfisol in the Dry Zone of Sri Lanka

Soil moisture is one of the major factors that determines volatilization of urea. In this study, we investigated the effect of soil moisture on urea volatilization from an Alfisol (Reddish Brown Earth soil). A bulk soil sample was collected from the research field of the Faculty of Agriculture, Rajarata University of Sri Lanka. A pot experiment was arranged under completely randomized design with four replicates per treatment. Soil moisture levels were maintained at 0 kPa (volumetric water content (VWC): 33%), 33 kPa (VWC: 16%), 44 kPa (VWC: 8%), and 54 kPa (VWC: 2%) as the treatments. A 50% overhead shade net was hung at 1.8 m aboveground covering the treatment pots. Urea was applied to each pot at a rate of 1,276.5 mg kg-1 and emitted NH3 was collected using the enclosure method. Soil NO3−-N, NH4+-N, pH, and EC were determined at six times at two-day intervals. The effect of soil moisture, time, and their interaction effect on NH3 volatilization was analyzed using repeated measure ANOVA with post-hoc Tukey HSD test for mean comparison. The NH3 volatilization significantly decreased with time (p < 0.05). Time and treatment interactions effect were significant for volatilized NH3 and other soil parameters measured except for soil NO3−-N (p < 0.05). A decreasing trend was observed for NH3 volatilization at 0 kPa, 33 kPa, 44 kPa & 54 kPa metric suctions as 6.3 g m-2 day-1, 4.9 g m-2 day-1, 4.7 g m-2 day-1, and 0.065 g m-2 day-1 respectively. Soil NH4+-N and EC fluctuated significantly among the treatments. Maintaining the soil moisture at 33 kPa during urea application is recommended for achieving high fertilizer use efficiency.

Synthesis of Pyridyl‐Imine Complex of Iron(III) as a Fluorescent Sensor for Hydrazine Detection Based on Redox Chemistry

AbstractA six‐coordinated iron(III) pyridyl‐imine complex was synthesized by one‐pot reaction and was demonstrated as a fluorescent sensor for the detection of hydrazine in solution based on the redox reaction. The fluorescence signal generated was attributed to the reduction of the oxidative iron‐center of Fe(III) to Fe(II) by the reductive hydrazine. A good linear relationship for the analysis of hydrazine was obtained with the concentration range of 0.06–0.15 mM (R2=0.99114) in DMF solution. The limit of detection (LOD) estimated was 14.8 μM and the limit of quantitation (LOQ) was 49 μM. Moreover, the color change of the iron(III) pyridyl‐imine complex solution from dark green to reddish brown was able to observed by naked eye, indicating that the Fe(III)‐complex may be also utilized as a colorimetric sensor for hydrazine at millimolar level. The present study demonstrated an interesting fluorescent sensing system based on redox chemistry of an iron(III) complex that generated fluorescence response upon reacting with its redox pair such as the reducing agent hydrazine.

Occurrence and Sustenance of Arbuscular Mycorrhizae in Selected Dry Zone Agricultural Lands in Sri Lanka

Arbuscular mycorrhizae (AM) are one of the important symbiotic micro-organisms that provide a primary natural pathway for soil/plant nutrient transfer. The prevalence of AM on Reddish Brown Earth soils in the dry zone was determined under three agronomic management regimes: least-, moderate- and well-managed at Kiralogama, Puliyankulama, and Medawachchiya, respectively, in the Anuradhapura District. Arbuscular mycorrhizae interactions were evaluated with varieties each; sesame (Sesamum indicum L.), bitter gourd (Momordica charantia L.), cowpea (Vigna unguiculata (L) Walp.), chilli pepper (Capsicum annuum L.) and tomato (Solanum lycopersicum L.), and locally-common four weed/non-crop species; akmella (Synedrella nodiflora (L.) Gaertn.), balathana (Eleusine indica (L.) Gaertn.), kurakkan (Eleusine coracana (L.) Gaertn.), and wal-rubber (Euphorbia heterophylla L.) in pot experiments. Rhizosphere soils at the well-managed Medawachchiya site recorded the highest number of average live (1312) and dead (2637) spores per 100 g of soil retained on 125 μm sieve compared to the lowest live (459) spores at the least-managed site at Kiralogama. At both Kiralogama and Puliyankulama, live spore counts significantly increased with the progress of Maha rainy season. Increases in plant P and Zn concentrations showed strong positive correlations with AM colonisation. Significant differences of both AM colonisation and spore counts were observed in pot experiments depending on the weed/non-crop species and the variety of crops. Akmella (51.2 %), bitter gourd (var. Black - 69.1 %), Sesame (var. Mali - 76.7 %) and wal-rubber (76.8 %) were associated with high levels of AM colonisation as well as soil spore counts.

Evaluation of soybean genotypes for resistance against the rust‐causing fungus Phakopsora pachyrhizi in East Africa

AbstractSoybean rust, caused by the biotrophic fungus Phakopsora pachyrhizi, is the most important foliar disease of soybean (Glycine max) worldwide. Deployment of resistant soybean cultivars is the best option for managing this disease. Genes conferring resistance to P. pachyrhizi have been identified, but pathotypes of the rust fungus overcoming these resistance genes have also been found. To identify novel resistance genes, soybean genotypes from both local and international sources were screened at multiple locations in Tanzania and Uganda in 2016 and 2017. The results from this screening revealed that infection types, disease severities, and sporulation levels varied among the genotypes and locations. The majority of the genotypes had tan‐coloured (TAN) lesions and developed moderate sporulation, implying susceptibility, while only seven of the 71 lines had reddish‐brown (RB) lesions and showed low disease severities in all of the screening environments. We identified seven genotypes that were the most resistant to rust in the most locations over the two years. These genotypes will be useful for further studies and, ultimately, for rust management, as they show broad resistance to various pathotypes of the rust fungus.

Effect of Foliar Application of Manganese, Zinc and Copper on Growth and Yield of Chilli (Capsicum annum. L)

Chilli (<i>Capsicum</i> <i>annum</i> L.) is an important spice crop grown in Sri Lanka. Deficiency of micronutrients, especially Mn, Zn and Cu reported in chilli growing areas has affected adversely on chilli production. Therefore, a study was conducted to evaluate the effect of foliar application of Mn, Zn and Cu on growth and yield of chilli grown in Reddish Brown Earth soils (Rhodustalfs), in dry zone, Sri Lanka. Seven treatments were tested in a greenhouse as pot experiment. Nitrogen, phosphorus and potassium were added into all seven treatments. Manganese Zn and Cu were added separately as foliar application for three treatments. One treatment had all three elements and control treatment had only N, P and K but no any added elements. Since all micronutrients were added as their sulfate form, last two treatments had S with N, P and K to assess the effect of S on plant growth. Copper with N, P, K added treatment showed significantly higher growth and yield compared to other treatments. It showed about 25% yield increment compared to control treatment. Therefore, it can be concluded that application of Cu is beneficial to obtain a better yield in chilli in Reddish Brown Earth Soils (Rhodustalfs). However, further field studies are needed to confirm findings.

Development of silver-polyvinylpyrrolidone nanocomposite for the selective and sensitive detection of sulfide from aqueous sample and its antimicrobial activity

The current study was focused on detection of sulfide ion colorimetrically form aqueous solution by using silver-polyvinylpyrrolidone nanocomposites (Ag–PVP NCs). The prepared NCs was analyzed by using UV–visible spectrophotometer, X-ray diffraction (XRD) analyzer, fourier transform infrared spectrometer (FTIR), transmission electron microscope (TEM), particle size analyzer, energy dispersive X-ray spectrometer (EDS) and zeta sizer. The minimum detection limit for sulfide by Ag–PVP NCs was calculated to be 1.05 nM. The decrease in absorbance and colour variation from brownish yellow to reddish brown was observed after the interaction of sulfide ion with Ag–PVP NCs due to the aggregation of NCs. The competitive ions presence with the analyte solution did not interfere the sulfide ion detection ability by prepared probe. The effective detection of the probe was studied with various pH, temperature and salinity. The proposed probe was found to be an efficient tool for the sensitive, selective, cost effective and on-site detection of sulfide ions from various aqueous media.

BIOMASSA HIJAUAN Mucuna bracteata DAN PENGARUHNYA TERHADAP KADAR N TANAH DI PERKEBUNAN KELAPA SAWIT

Mucuna bracteata is one of the legume cover crop that is usually used in oil palm and rubber plantations. In general, legume cover crop are planted during the preparation / after land clearing when the oil palm are planted, Mucuna bracteata has covered the planting area. Mucuna bracteata growth conditions will affect soil biological properties, namely organic nutrient content, C/N balance, microbial population and macro-fauna activity. This study aimed to describe the soil and Mucuna bracteata plants in the phase of oil palm plants. The study was conducted in April - June 2019 in the area of ​​TBM 3 oil palm plantations in 2 locations, namely Pabatu Plantation and Tinjowan Plantation. Soil types in Pabatu Plantation are typic dystrudepts (Reddish Brown Podsolik) and in Tinjowan Plantation the type of soil is typic hapludults (Yellowish Red Podsolik) and some typic paleudults (Yellow Podsolik). This study used a descriptive design that describes the growth character / biomass of Mucuna bracteata on TBM 3 oil palm plantations. The results of this study are the fresh biomass of Mucuna bracteata in Pabatu Plantation is 43,260 kg / ha and Tinjowan Plantation is 34,146 kg / ha (79%) and dry biomass is 13,943 kg / ha in Pabatu Plantation and 12,705 kg / ha in Tinjowan Plantation. The levels of N leaves of Mucuna bracteata are in the high category. Soil organic matter content and total N in Pabatu and Tinjowan Plantation are in the low category.

First Report of Lasiodiplodia theobromae Causing Stem Canker of Fraxinus americana

First Report of <i>Lasiodiplodia theobromae</i> Causing Stem Canker of <i>Fraxinus americana</i>

A Rice Variety ‘Tocomi-1’ with High Tocopherol Content and Reddish Brown Pericarp

A Rice Variety ‘Tocomi-1’ with High Tocopherol Content and Reddish Brown Pericarp

Reddish Brown Rice Variety ‘Superhongmi’ with High Taxifolin Content

Reddish Brown Rice Variety ‘Superhongmi’ with High Taxifolin Content

Demystifying the Stinking Reddish Brown Stains Through the Dermoscope: Cydnidae Pigmentation.

Demystifying the Stinking Reddish Brown Stains Through the Dermoscope: Cydnidae Pigmentation.

Marker-assisted pyramiding of four QTL/genes for Asian rust (Phakopsora pachyrhizi) resistance in soybean

ABSTRACTGene pyramiding (assembling multiple desirable genes into a single genotype) via conventional methods to combat Asian rust resistance in soybean (Glycine max (L.) Merrill) and developing soybean varieties with durable resistance to this disease continues to be a challenge. Therefore, our objectives were to pyramid four Asian rust (caused by Phakopsora pachyrhizi) resistance (Rpp) genes, viz., Rpp1, Rpp2, Rpp3, and Rpp4 from their respective donors, PI200492 (Komata), PI230971, PI462312 (Ankur), and PI459025 (Bing Nan). Two single crosses (PI200492 × PI230971; PI462312 × PI459025) and a double-cross [(PI200492 × PI230971) × (PI462312 × PI459025)] were made for present study. The scoring of parents and crosses was done according to a 0–9 grading scale, where 0 = 0% disease intensity (absolutely resistant), 1 = 1% disease intensity (highly resistant), 3 = 1.1–10% disease intensity (moderately resistant), 5 = 10.1–25% disease intensity (moderately susceptible), 7 = 25.1–50% disease intensity (susceptible), and 9 = more than 50% disease intensity (highly susceptible). Out of four parents used, Rpp1 gene-donor plant introduction (PI)200492 (Komata) was immune to rust, with a disease grade of 0.92. Rpp2 and Rpp4 gene donors, PI230971 and [PI459025 (Bing Nan)], had reddish-brown (RB) lesions. Rpp3 gene-donor PI462312 (Ankur) showed a few, highly localized patches of tan lesions; however, it was not as susceptible as the susceptible check JS335. In cross-A (PI200492 × PI230971), all the F1s were rust resistant, whereas in the F2, 93.9% (845 plants) were either immune (11.0%, 99 plants) showing no infection or had RB lesions (82.9%, 746 plants) with resistant reaction and the remainder 6.1% (55 plants) had susceptible reaction. In cross-B (PI462312 × PI459025), 80.8% (63 plants) of the F1 had RB lesions and resistant reaction, whereas 19.2% (15 plants) had susceptible reaction. In its F2 population, 24.2% (217 plants) were susceptible. In cross-C [(PI200492 × PI230971) × (PI462312 × PI459025)], i.e., double-cross hybrid (DCH), 85.5% (77 plants) were either immune or had RB lesions, whereas the rest of the plants 14.5% (13 plants) showed susceptible reaction. Based on parental survey, four simple sequence repeat primers were short-listed for amplification of individual rust-resistance gene-specific markers in ten plants of each cross, viz., Satt191-210bp (Rpp1 gene), Sat_361-245bp (Rpp2 gene), Satt263-195bp (Rpp3 gene), and Rpp4TM-128bp (Rpp4 gene). In the single-cross hybrid (SCH)-A (PI200492 × PI230971), six plants had both Rpp1 and Rpp2 genes, whereas in another SCH-B (PI462312 × PI459025), seven plants had both Rpp3 and Rpp4 genes. In DCH [(PI200492 × PI230971) × (PI462312 × PI459025)], a single plant had all four Rpp genes, five plants had two Rpp genes, whereas a single susceptible plant had only Rpp3 gene. The homozygous rust-resistant segregants from these crosses can be used for improving rust resistance of otherwise adapted, high-yielding soybean genotypes.

Identification of stingless bees (Hymenoptera: Apidae) in Kenya using morphometrics and DNA barcoding

Stingless bees are important pollinators of wild plants and crops. The identity of stingless bee species in Africa has not been fully documented. The present study explored the utility of morphometrics and DNA barcoding for identification of African stingless bee populations, and to further employ these tools to identify potential cryptic variation within species. Stingless bee samples were collected from three ecological zones, namely Kakamega Forest, Mwingi and Arabuko-Sokoke Forest, which are geographically distant and cover high, medium and low altitudes, respectively. Forewing and hind leg morphometric characters were measured to determine the extent of morphological variation between the populations. DNA barcodes were generated from the mitochondrial cytochrome c-oxidase I (COI) gene. Principal Component Analysis (PCA) on the morphometric measurements separated the bee samples into three clusters: (1) Meliponula bocandei; (2) Meliponula lendliana + Plebeina hildebrandti; (3) Dactylurina schmidti + Meliponula ferruginea black + Meliponula ferruginea reddish brown, but Canonical Variate Analysis (CVA) separated all the species except the two morphospecies (M. ferruginea reddish brown and black). The analysis of the COI sequences showed that DNA barcoding can be used to identify all the species studied and revealed remarkable genetic distance (7.3%) between the two M. ferruginea morphs. This is the first genetic evidence that M. ferruginea black and M. ferruginea reddish brown are separate species.