نشریه رستنیها
سال بیست و چهارم شماره 2 (پیاپی 69، Autumn and Winter 2023)

The protection of genetic resources is crucial for safeguarding national assets as natural resources face various threats such as climate change and changing land use. To conserve endangered species such as Astragalus fridae, an endemic species of gypsum soils of Semnan province (Iran) which is on the brink of extinction due to limited distribution and habitat destruction, establishing a gene bank is essential. To ensure the survival of this species, it is necessary to extract high-quality and high-quantity DNA. The present survey, has evaluated five genome extraction methods including Protocol A (main CTAB), Protocol B (CTAB without β-mercaptoethanol), Protocol C (CTAB without ammonium acetate), Protocol D (the modified Murray & Thompson method), and Protocol E (GeneAll Plant Kit), to determine their ability to extract DNA from fresh and herbarium leaves of A. fridae. The quality and quantity of DNA were assessed using gel electrophoresis, spectrophotometry, and usability of the purified DNA tested by PCR of the ITS region of nuclear ribosomal DNA. Protocol D yielded the highest quality of the extracted genome from fresh leaves, and Protocol A, extracted the highest DNA concentration from fresh leaves. Although Protocols B and C extracted a reasonable amount of genome of acceptable quality with dry leaves, the ITS region was not amplified in these samples, rendering them unsuitable for DNA extraction of A. fridae.

During collecting and identifying of Xylariales fungi (Ascomycota) of Iran, a specimen was collected from Siahkal forest (Guilan Province, northern Iran) in autumn 2021. Based on morphological and molecular (ITS sequence) data, this fungus was identified as Cryptosphaeria multicontinentalis, which is the first report for the funga of Iran. Detailed taxonomic and phylogenetic information related to this species is described and illustrated. In addition, an updated list of species of Diatrypaceae is provided from Iran.

Pollen grains of eight species of Gagea were examined using Light Microscope (LM) and Scanning Electron Microscope (SEM). Results showed that selected quantitative characters of pollen grains (polar axis, equatorial axis, P/E ratio, width of muri, lumina or pore number per 2 μm2 in the exine surface, and lumina or pore size) show significant variation providing potential diagnostic characters in discrimination of taxa in the genus. Among the pollen qualitative characters, pollen shapes varied from oblate to oblate-spheroid, and pollen size can be classified as medium (less than 50 μm) and large (more than 50 μm). Exine ornamentation in equatorial view and on sulcus margin region show also variation among the studied taxa. Cluster analysis of selected pollen morphological characters indicated two pollen groups and three types. The palynological types supported taxonomical situation of G. sect. Didymobulbos, G. sect. Plecostigma, and G. sect. Platyspermum. There is no pollen type for studied taxa of G. sect. Stipitatae. In addition, palynological issues suggest a close relationship between G. sect. Didymobulbos and G. sect. Stipitatae.

During an investigation on endophytic fungi of grasses of the tribe Triticeae (subtribe Triticinae) in the west and northwestern provinces of Iran, two species, viz. Sarocladium terricola and Selenophoma linicola, and one genus, Septoriella (S. allojunci), were newly identified for the funga of Iran using morphological traits and sequences of the internal transcribed spacer regions 1 and 2 including the intervening 5.8S nuclear ribosomal DNA (ITS). Selenophoma linicola and Septoriella allojunci, members of the class Dothideomycetes, were isolated from stem of Aegilops cylindrica and leaf of A. cylindrica in W. Azerbaijan and Lorestan provinces, respectively. Sarocladium terricola, belonging to the Sordariomycetes, was isolated from leaf of Triticum aestivum in Kermanshah province. All identified species are described and illustrated herewith, and compared with closely related species. Aegilops cylindrica and T. aestivum are reported as new hosts (matrix nova) for these fungi.

Salvia multicaulis is a widespread medicinal plant growing in north, northwest, and center of Iran. In this study, the aerial parts of 29 specimens belonging to five geographical populations were collected and dried at full flowering stage. AFLP molecular markers were applied for genetic diversity analysis. Essential oil was extracted by hydro-distillation and its chemical compositions were analyzed by GC/MS. AFLP results led to the recognition of two different genotypes. The results demonstrated high molecular affinity among Azerbaijan and Kurdistan populations. However, Tehran showed much more correlations with Isfahan and Kohgiluyeh-o-Boyerahmad populations. The AMOVA test results, demonstrated high within population genetic variability. STRUCTURE analysis revealed some degree of population genetic fragmentation that was principally due to genetic difference occurred between Azerbaijan and the rest of the studied populations. Overall, 40 constituents were determined among which, a-pinene (11.2–19.9%) and 1,8-cineole (10.2–20.4%) were distinguished as two major compounds. Tehran showed closer affinity to Isfahan and Kohgiluyeh-o-Boyerahmad. The results of molecular studies were in accordance with those obtained by the chemical compositions. Therefore, ecological factors and geographical distance as well as genetics play an important role in the species evolution and distribution in different parts of the country.

The present study is focused on identifying the differences between the closely related sections Microphysa and Campylanthus in the genus Astragalus using micromorphological characteristics. For this purpose, the morphological characteristics of leaf epidermal cells and stomata as well as pollen grains have been studied. The results obtained from light- and scanning electron microscopy (LM and SEM) indicated that, stomatal type was typically anemocytic in most species, while the anisocytic type was only found in two species, viz. A. callistachys and A. microphysa. The epidermal cells in most species were observed to have a polygonal shape with straight anticlinal walls. However, in A. cephalanthus, both polygonal and irregular cell shapes as well as sinuate anticlinal walls were observed. All pollen grains shed as monad and exhibited small size, single shape, tricolporate aperture type, isopolarity and symmetry, with rounded or elongated pores. It was concluded that, the micromorphological data alone were not enough to separate these two sections and determine the limits of the species.

In Flora Iranica area, Tanacetum comprised 18 sections and 54 species. Considering new findings, Tanacetum species in Iran has been increased to 37. No detailed information on the karyotype analysis of the studied species was found in the literature. Therefore, in the present study, the detailed karyomorphology of six populations belonging to five taxa of Tanacetum sect. Xanthoglossa were studied. The basic chromosome number of the Tanacetum has been reported to be x=9. Mitotic chromosome number of studied taxa were found to be diploid (2n=2x=18) for T. argyrophyllum, T. oligocephalum (Khoy), T. uniflorum (no previous karyological records are known for this species) and T. pinnatum. According to the symmetry classes of Stebbins, class 2A was found predominant karyotype class among the studied Tanacetum except for tetraploid (2n=4x=36) T. polycephalum subsp. heterophyllum which was placed in 2B. Karyotype analysis of each taxon has been presented and discussed.

Sphaerulina is a genus of fungi which belongs to the family Mycosphaerellaceae in the order Mycosphaerellales. In a survey of leaf-inhabiting fungi associated with trees in Hyrcanian forests, several isolates of the genus Sphaerulina were recovered from leaf spot symptoms of oak trees. Integration of multi-locus phylogenetic analyses (LSU, ITS, actA and tef1) and morphological identification were applied to identify the isolates. The results revealed that, the isolates belong to a species of the genus Sphaerulina, namely, S. quercicola. To the best of author’s knowledge, this is the first report of this species for the funga of Iran.

An isolate of an ascomycetous fungus collected during a field trip in the Zagros Forest, Kermanshah province (West of Iran) was identified as Myxotrichum. Based on the morphological and molecular characters the isolate was identified as M. deflexum. Genomic DNA was extracted and a nuclear rDNA region, containing the internal transcribed spacers 1, 2 and 5.8S gene of rDNA (ITS) were amplified and PCR product was sequenced. Amplicon was purified, sequenced and submitted to the GenBank (Acc. No. OR047187). The BLAST search results showed highest similarity of this isolate to other isolates of M. deflexum from GenBank. By reviewing the literature on ascomycetous fungi, Myxotrichum is found as a new genus for the funga of Iran.

In the winter of 2023 and during routine examinations of date palm trees in Safiabad, Dezful (Khuzestan Province, Iran), an intense white mass of fungal molds was observed on the leaf scars of the said trees. Purified isolates were subjected to morphological and molecular examinations. Typical morphological characteristics viz. single-celled, hyaline, smooth and obovate to ellipsoidal conidia arising by sympodial growth with basal scar prominent produced holoblastically on denticulate conidiogenous cells revealed that, all fungal isolates belong to the genus Sporothrix. Furthermore, based on a combination of morphological and sequence data of internal transcribed spacer (ITS) of rDNA, the recovered fungal isolates were determined as S. ranii. Therefore, based on the present information, it is the first report of this species for the funga of Iran and its second report in the world.