نشریه رستنیها
سال بیست و یکم شماره 2 (پیاپی 60، پاییز و زمستان 1399)

Lilium ledebourii is a rare Asian species and is an endemic plant of Iran which grows in Gilan province in Damash highlands. In this study, the structural features of flower including the study of sections such as carpel, stamen, and tepals were performed. This plant had a full flower that consisted of six tepals (three petals and three petal-like sepals), six free stamens and a 3-lobed gynoecium. In the cross-section of tepals, the stomata as well as cells containing anthocyanins were observed. In stamens, anther (epidermis, endothecium, middle layers and an innermost tapetum), and filament were observed. In the gynoecium, ovary (ovary wall, locule, funiculus, anatropous ovule, different arrangements of vascular bundles, transmitting tissue and stomata), style (stylar canal and transmitting tissue), and dry stigma with elongated papillae tissue were observed. Besides, the presence of alkaloids in the bulb was tested using Wagner and Dragendorff reagents and the total alkaloid content in three types of methanol, ethanol, and butanol extraction was investigated. The ethanol extract had a significant difference with other extractions. The methanol extract was chosen for further investigation by gas chromatography-mass spectrometry (GC-MS) and two compounds of Pterin-6-carboxylic acid, and Pyrrolizin-1,7-dione-6-carboxylic acid, methyl (ester) were detected.

Anemone L. (Ranunculaceae) with ca. 150 morphologically variable species is distributed all over the world. Morphological characters are widely used to discriminate species. In the present study, therefore, the Iranian species of Anemone and Pulsatilla are investigated by their achene macro- and micro-morphological and anatomical features. The micro-morphological and anatomical studies are performed employing the scanning electronic microscope (SEM). Multivariate statistical analyses of various quantitative and qualitative characters of the achene as well as their anatomy, resulted in distinguishing four different achene types. Although, obvious differences are observed in the achene body surface; number of the endocarp and mesocarp layers, and in the shape and length of the style, it is also revealed that, features like the number of epicarp layers and margin surface, have no diagnostic values. The analyses are carried out by SPSS software, using the Ward’s and the Average linkage methods. Both the dendrograms showed that, the P. albana is separated from other species which is consistent with systematic studies and the achene features. Besides, in the present study, A. blanda is also rediscovered.

Alhagi is a small genus of desert or semi-desert shrubs with about six species widely distributed throughout northern Africa, eastern Mediterranean, the Middle East to the Central Asia, and small parts of India and China. Despite previous molecular and morphological studies on the genus, the taxonomy of Alhagi has been remained controversial. In order to delimit natural groups as well as to clarify interspecific relationships of Alhagi species distributed in Iran, 45 ingroup taxa representing 30 accessions of the genus Alhagi and 15 species from the sister genera, as well as two species, namely, Astragalus arthuri and Colutea persica as outgroups were selected. The nuclear ribosomal DNA internal transcribed spacer was employed for phylogenetic reconstructions. Phylogenetic analyses were performed using maximum parsimony, maximum likelihood and Bayesian inference. The obtained results recovered Alhagi accessions as a well-supported monophyletic group. All three traditionally recognized species in Iran split into two distinct morphologically well-supported lineages similar to splits graph. The results showed that, Alhagi maurorum and A. graecorum are recognized as two distinct species distributed in two phytogeographical regions of Irano-Turanian and Saharo-Sindian. As a complementary taxonomic treatment, the diagnostic key and description of the species are also provided herewith.

The classification of the genus Salixis troubled by the intra-specific flexibility and diversity of the morphological features. The aim was to prepare a comparative description of petiole anatomy of 23 species of Salix, in addition to two species of Populus and Bivinia jalbertii, as well as to determine the taxonomic importance of petiole features. In order to do so, 10 quantitative and 10 qualitative petiole anatomical features were studied. Moreover, a Bayesian analysis was executed on the relevant ITS sequences obtained from GenBank, included 28 specimens from 17 species. The anatomical features were then mapped on the resulted cladogram and the ancestral states of some prominent petiole anatomical features were reconstructed. Based on the results, no stomata were observed in the petiole's epidermis of any of the species studied; therefore, it could be assumed that, the petiole in the studied species is dependent to the leaf blade for respiration and gas exchange. While the quantitative petiole anatomical features were turn to be variable in the individuals of the same species grown in different habitats, qualitative features showed a desirable consistency. The petiole anatomical features studied were proved to be useful in the differentiation and identification of taxa at sub-generic and specific levels. In addition, the reconstruction of the ancestral state of the studied features revealed that, polygonal cell shape, straight or slightly curved anticlinal wall pattern, horizontal compression of the petiole, the presence of lateral ribs, convex adaxial surface and ribbed petiole outline were primitive features in the species studied.

The dry bubble disease (caused by Lecanicillium fungicola) is the most threatening disease of cultivated button mushrooms throughout the world. Most of the symptoms shown by the dry bubble are also shown by the wet bubble (Mycogone perniciosa) and they may even be confused. Early detection and precise monitoring of the diseases are very important for managing effective treatments. A PCR-based assay for a specific diagnosis of the disease in various stages of the disease development was developed. A primer pair designed based on ribosomal DNA generated two bands, 650 and 800 bp, specific for the detection of L. fungicola. Additionally, a simple and time-saving method for direct extraction of DNA from the affected sporophores is presented and the impact of sample pre-treatment on the efficiency of DNA isolation is highlighted. Here, we also report Simplicillium lamellicola from Iran, using the nuclear ribosomal internal transcribed spacer (ITS) region.

Pure DNA is essential in various techniques of molecular biology and its extraction from plants to produce large amounts of secondary metabolites is a difficult task. Alchemilla is known to synthesize a large number of secondary metabolites which reduce the quality of the extracted DNA. This study, aimed to set up a method for high-quality DNA isolation from Alchemilla leaf. We examined three extraction methods. Furthermore, a comparison concerning price, simplicity, and security is carried out. We optimized a CTAB-based method using increasing the volume and concentration of CTAB buffer, lysis time, and cold incubation period, performing six times dilutions, and three times precipitations, adding polyethylene glycol, and removing toxic or expensive materials. The results showed that, 260/280 and 260/230 ratios of extracted DNA by the optimized method with the concentration of 595–387 ng/µL were 1.75–1.82 and 1.56–1.68, respectively. The quality of extracted DNA by this method was significantly higher (p < 0.001) than that of other ways, so that all samples were positive for DNA, as assessed by electrophoresis and PCR. The optimized method was simple, effective, reproducible, relatively non-toxic, and inexpensive. The results revealed that, this method was successful in producing large amounts of high-quality amplifiable DNA.

As a highly valuable source of ephedrine, Ephedra major is well-adapted to various habitats in Iran. Over the last few decades, human’s activity, in particular, over exploitation of its populations to extract medicinally valuable constitutes, have endangered this species. In this investigation, the level of genetic diversity and genetic structure in 16 natural populations of E. major for the very first time have been assessed utilizing molecular markers, inter-simple sequence repeat (ISSR). A high level of genetic diversity was revealed among the population where Dehbar region (Khorasan Razavi province, Iran) exhibited the highest quantity of genetic parameters (P = 67.96%, H = 0.365 236, I = 0.365). Intra-population genetic diversity (71%) was significantly higher than inter-population genetic diversity (29%) and only 1% genetic diversity among-regions. The unweighted pair group method with arithmetic mean (UPGMA) and Bayesian clustering methods indicated the presence of a relatively strong population structure and to some extent in accordance with the geographical origin of populations. However, the success of UPGMA in grouping populations in congruence with their geographical location was more tangible. From the outcomes of this study, it could be asserted that, ISSR markers are effective in detecting the pattern of genetic diversity in E. major populations. On the other hand, by having reliable knowledge of the status of genetic diversity and structure of populations of this species, the first steps can be taken to protect populations with low genetic diversity, and also pave the way for breeding programs to finding and selection of superior populations.

Teucrium polium is a Mediterranean native subshrub with large distribution range and high medicinal values which overexploitation of its population has become a serious danger and conservation attempts is needed. Having the genetic information of its population can help to clarify the complicated phylogenetic relationships of species within the sections, genetic conservation and establishing breeding programs to develop cultivars and ultimately preventing declining the populations of T. polium. The genetic diversity parameters and populations structures of 16 populations of T. polium at the local scale in the Alborz mountain range of Iran were assessed using inter-simple sequence repeats (ISSR) primers. The averages of polymorphism (P%), Nei’s genetic diversity (H), and Shannon’s Information Index (I) were 33.24%, 0.118 and 0.179, respectively. The population of Asara to Gach-Sar presented the highest P%; 43.28%, H; 0.163 and I; 0.243. AMOVA analysis indicated that, a large portion of genetic variation as within population (77%), and a relatively high genetic differentiation (Gst: 0.311) and gene flow (Nm: 1.107) among populations were observed. UPGMA tree and PCoA plot of ISSR data divided the populations into three genetic groups to a significant extent based on the geographical origins. Similarly, the results showed that, STRUCTURE analysis grouped the populations into three clusters with significant geographical affinity. Teucrium polium exhibited a strong structure and genetic differentiation with low to moderate genetic diversity.

The current study was conducted to explore genetic variations among several cotton species, including Gossypium hirsutum cultivar Varamin, G. barbadense cultivar T14, G. herbaceum cultivar Arya, hybrid of G. hirsutum cultivar Varamin × G. barbadense cultivar T14 (V×T14), and a hybrid of G. hirsutum cultivar Varamin × G. herbaceum cultivar Arya (V×A) using inter-simple sequence repeats (ISSR). In this study, a total of 60 bands were amplified using four ISSR marker primers. As a result, 57 bands showed polymorphism. The results showed a high level of diversity in the studied varieties. The highest percentage of polymorphism was related to ISSR4 primer with ACACACACACACACACT sequence (100%) and the lowest percentage was related to ISSR1 primer with AGAGAGAGAGAGAGAGC sequence (92.8%). Cluster analysis showed three major groups. The first group consisted of Arya and Varamin, the second group was comprised of V×A, T14 which showed a high genetic similarity indicating T14 originates from Varamin cultivar, and the third group contained V×T14. Overall, the results showed that, ISSR markers can be effectively used to study the genetic diversity of cotton.

To clarify taxonomic complexity within the Anthemis species, pollen grains of 19 species belonging to three sections of the genus Anthemis sensu strictoand two species of the genus Cota from Iran were examined by using scanning electron microscopy. The pollen grains morphological characteristics are provided for this genus in detail. Pollen grains were trizonocolporate and echinate. The pollen grains shape was prolate and spheroidal with the mean polar axes 20–35 μm and the mean equatorial diameter 20.3–26.5 μm. Among the studied taxa, A.microcephala showed the smallest pollen grains, and A. odontostephana possessed the largest ones. The spines were commonly conical with a broad basis, ornamentation within spines was generally rugulate-perforate. Thirteen pollen quantitative and qualitative characters have been analysed statistically by employed UPGMA, PCA and MDS in PAST software. Numerical analysis showed that, features as pollen shape, exine ornamentation, spine length and density were valuable characters for separating the examined taxa. In the resulting clusters, three groups within the investigated species have been recognized. The results derived from the present study, therefore, proved to be concomitant with the previous study on taxonomic infrageneric classification. The palynological evidences confirmed the sectional divisions, and revealed that, A. brachystephana and A. lorestanica in A. sect. Anthemis, have close relationship with A. odontostephana. The palynological characters of sect. Anthemis were similar to Tripleurospermum and Cota. The analysis of the palynological data confirmed the conventional taxonomic classification of the genus Anthemis rather than the phylogenetic classification of the genus.

The Pezizales are an order of the subphylum Pezizomycotina within the phylum Ascomycota. Members of this order are characterized by unitunicate asci that typically open by rupturing to form a terminal or eccentric lid or operculum (Hansen & Pfister 2006). Of the 1638 Pezizales species known so far, Geopora Harkn. (Pyronemataceae) is represented by 23 species (Kirk et al. 2008). Geopora spp. are characterized by entirely or partially hypogeous, globular, semi-globular or cup-shaped ascocarps, whitish, greyish or yellowish grey hymenium, cylindrical, 8-spored and operculate asci, generally bifurcate, septate and hyaline paraphyses, ellipsoid, smooth ascospores mostly with one or two larger oil drops and some smaller oil drops (Tamm et al. 2010, Perić & Perić 2011). The members of the Geopora spp. are widely distributed in the Northern Hemisphere. Many researchers have reported Geopora species in North America, central, north, south and southeast Europe, and the Middle East (Tamm et al. 2010, Ashraf & Khalid 2012, Guo et al. 2019, Saba et al. 2019, Uzun & Kaya 2019). Saber & Pegler reportedthe presence of G. arenicola (Lév.) Kers in Iran as Sepultaria arenicola (Ershad 2009). In this study, two specimens of G. cooperi associated with Pinus nigra were collected in Khorassan Razavi province (Iran) in 2020 (Fig. 1a) with the following characteristics: Ascomata 50–55 mm in diameter, globose, subglobose to oblong, irregularly convoluted, peridium dark brown and covered with light brown hairs, gleba white to pinkish-white, highly convoluted, convolutions close but separate (Fig. 1b, c). Hymenium 200–250 μm thick. Asci 180–270(215) × 15–22(16) μm, cylindrical, 8-spored, non-amyloid and tapering at the base (Fig. 1d, e). Paraphyses cylindrical, septate, swollen at the tips, nearly of the same level as ascus, enlarged at the apex up to 11.5 μm (Fig. 1f). Ascospores 18–27(23) × 13–17(14) μm, smooth, hyaline, broadly elliptical, thin-walled, generally with a large central guttule (Fig. 1g, h). Ectal excipulum 120 μm, 4–5 cells thick, dark brown, textura globulosa,cells oblong to circular in section, smooth, walls thick and dark brown, 24–40 × 18–27 μm, cells on the outside darker than the inner cells (Fig. 1i). Excipular hairs hyphae-like, branched, very long to 1000 μm, 5–17 μm wide, blunt at the tips and dark walls (Fig. 1j). All these characteristics were consistent with other authors’ descriptions for G. cooperi (Tamm et al. 2010, Guo et al. 2019).While Geopora cooperi isconsidered as a very rare species by Perić & Perić (2011), itwas widely reported from Europe (Austria, Sweden, Norway, Denmark, Switzerland, Spain, and Italy) by Nannfeldt (1946), Moser (1963), Burdsall (1968), Soleilhac (1972), Ortega et al. (1981), Moreno et al. (1991), and Montecchi & Dal Forno (1995). This species is hereby reported new to the mycobiota of Iran. The mycorrhizal association of Geopora species with various conifers and deciduous trees such as Pinus ponderosa Lawson & Lawson (Fujimura et al. 2005), Pinus edulis Engelm., Cercocarpus ledifolius Nutt., Salix linearistipularis (Franch.) K. S. Hao, Quercus garryana Douglas ex Hook. (Frank et al. 2009), Quercus douglasii Hook., and Arn. (Smith et al. 2006), Picea abies (L.) Karst.(Tedersoo et al. 2006), Epipactis atrorubens (Hoffm.) Besser. (Shefferson et al. 2008), Abies grandis (Dougl.) Lindl., Cedrus spp., and Populus spp. (Tamm et al. 2010) have been confirmed by molecular techniques. Fujimura et al. (2005) have reported G. cooperi form mycorrhiza with Pinus ponderosa. Tedersoo et al. (2006) have identified ectomycorrhizal Geopora spp. on root tips both in coniferous and deciduous woodlands. They also have found Geopora spp. occurred in various early- and late-successional woodlands in acidic to alkaline forest soils, in addition to cinder and burnt soils. Association of G. cooperi with P. nigra, has been previously reported by Burdsall (1968) from the USA. Specimen examined: Iran: Khorassan Razavi province, under Pinus nigra, solitary, on ground, 24 July 2020 (IRAN 16696F).

Topography has a great influence on flora and mycobiota of high-altitude regions. Iran is one of the most diverse countries in Western Asia having mountainous regions with several peaks above 4000 meters across the country. Flora and mycobiota of high altitude environments are specific because of living conditions and microclimate at high elevations. During recent botanical survey in Central Alborz mountain range a specimen of Noccaea stenocarpa (Boiss.) Al-Shehbaz (Brassicaceae) was collected from elevation of 2650 m. Almost all the leaves on collected plants were infected by an unknown Puccinia species with the following morphological features: Telia small, round, exposed, chestnut-brown, compact, mainly hypophullous, covering almost the whole leaf surface, occasionally on upper side of leaves. Teliospores two-celled, 30–50 × 11–19 µm, clavate, oblong-clavate, ellipsoid or oblong-ellipsoid, attenuated above and below, sometimes round or notched at apex, constricted at septum, wall smooth, hyaline to yellowish-brown, mostly darker at apex, 1–1.5 µm at sides, 2–6 µm at apex, pedicel hyaline, persistent up to 65 µm long (Fig. 1). These morphological features fit well with the earlier description of Puccinia thlaspeos Ficinus & C. Schub. (Ulyanishchev 1978). Puccinia thlaspeos is a Lepto-Puccinia, which means teliospores germinating in sori during the growing season. As this rust accompanying its host mostly in high-altitude environments, it should be considered as a montane rust (Abbasi & Hedjaroude 2002). To our knowledge this is the first report of P. thlaspeos from Iran. Material examined: Iran: Alborz province, Karaj-Chalus road, Dizin road, Varang Rud, on Noccaea stenocarpa (Brassicaceae), 2650 m, 24.5.2020, leg. A.H. Pahlevani (IRAN77285), III.

In recent field studies, Solanum viarum Dunal(Solanaceae) was collected and reported for the first time for the flora of Iran from around Qadikola in Mazandaran province (north of Iran). Notes on the morphology, taxonomy, geographical distribution, habitat and uses of this species are provided herewith. This is the first spiny species of the section Acanthophora in Iran. Solanum viarum Dunal in DC., Prodromus systematis naturalis regni vegetabilis 13(1): 240 (1852); Type: Sao Paulo. Brazil, Lund 799 (GDC. holo. IDC microf. 2080: 1.1). Plants mostly shrubs, 1–2 m tall, many-branched from the base; stems, petioles, pedicels, and leaves densely glandular puberulent, straight covered with hairs, hairs less than 0.5 mm long; stem with few prickles, stems and leaves with yellow spines up to 1.5 cm long; leaves simple, 7–14 cm long and 5–10 cm wide, margins lobed, usually with 3–5 pairs of shallow obtuse or acute lobes; petioles 3–9 cm long, with long prickles; inflorescence lateral, with 2–5 flowers; peduncles stout, 15–20 mm long; calyx pubescent on outside, 4–7 mm long; corolla white or greenish white, star-shaped with five petals, 15–25 mm diameter; fruits globose berry, up to 15–30 cm in diameter, light green mottled, dark green when immature, yellow at maturity, with a whitish or greenish layer; seeds 100–300 per berry, broad kidney-shaped, 2–3 mm in diameter, light red-brown (Fig. 1). The plant was identified and described by the help of studying the reliable flora and articles (Mullahey et al. 1993, Welman 2003, Waggy 2009). Specimen examined: Iran: Mazandaran province, 8 km south of Ghaemshahr, Qadikola forest, 100 m, Eskandari & Abdi (IRAN-77431). According to the Flora Iranica (Schoenbeck-Temesy 1972), 12 species, and Khatamsaz (1998), 10 species of Solanum are reported from Iran but based on a recent taxonomical and molecular study of Solanum (Eskandari et al. 2020), only eight valid species is reported for this genus from Iran. In general, accurate determination of the genera and species in the family Solanaceae in Iran, requires attention by performing more molecular study to clarify its actual taxonomic status (Ghahremaninejad & Riahi 2020). Solanum viarum grows originally in southern Brazil, Paraguay, Uruguay, and northern Argentina, but has also spread in South America, Central America, the West Indies, and the southern United States (tropical soda apple). This plant has long been naturalized in part of China and the Indian subcontinent (Babu 1971), and is reported in the Democratic Republic of the Congo and Cameroon (Jaeger & Hepper 1986) also. The species was accidentally and sporadically introduced into Africa and can be expected to spread to many other subtropical areas (Nee 1991), but recently becoming an invasive species in cattle pastures in the south-eastern United States. Since its introduction into the U.S., it has rapidly infesting to an estimation of one million acres of pastures, citrus groves, sugarcane fields, ditches, vegetable crops, forestlands (oak hammocks and cypress heads), natural areas etc. (Charles et al. 2012). In more tropical areas, flowering and fruiting occurs throughout the year (Nee 1991) while in open woodland and pine plantations of southern Africa, flowering season is confined to November to April with fruiting in August (Welman 2003). The root system of this plant consists of a deep and strong wooden base with numerous resistant lateral branches which under suitable conditions (low elevations mostly below 1000 m), can become invasive with a wide range of distribution. It is often a common weed of natural areas, improved pastures, croplands, roadsides and fence rows, waste places, open disturbed sites, secondary growth and forest edges (Waggy 2009). Solanum viarum contains chemical constituents like Solasodine which is found in many plants of Solanaceae family. The Solasodine has many medicinal properties and are used as fever-reducing agent, pain-reliever, laxative, diarrhea, anticoagulant and anti-asthma, anemia, burns, carminative, constipation, cough, cut restoration, food digestion, infections, mouth disinfection, skin diseases, toothache etc. in Iran. (Eskandari et al. 2019).