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Orkide yumruları (tuber) tarımsal üretimde gün geçtikçe değerlenmekte olup, ayrıca tıbbi drog önemine sahiptir. Ekonomik değerde olan bu yumrularla ilgili ontogenik bir araştırma bulunmamıştır. Bu nedenle; çalışmada orkide yumru ontogenisinin incelenmesi amaçlanmıştır. Materyal olarak Himantoglossum robertianum stolonlarından Historesin kiti ile gömme preperatlardan mikrotom kesitleri alınmış ve Toluidin Blue ile boyanmıştır. Sonuçlara göre; yumrunun alt tarafında statosit hücreleri ve kaliptranın olduğu ve yeni gelişecek olan yumrunun tüm yüzeyinin meristematik hücrelerle çevrili olduğu tespit edilmiştir. Yumrunun alt tarafındaki mersitemlerden ileride meristele, müsilaj ve nişasta hücreleri farklılaşırken, yumrunun yanal meristemlerinden ise sadece müsilaj ve nişastalı hücreler gelişmektedir. Yumrunun alttan yukarıya doğru hücrelerinde farklılaşma ile birlikte büyüdüğü ve aralarında meristeleler ile topaç şeklinde bir yapıya dönüştüğü tespit edilmiştir. Ayrıca floemden sakkaroz yüklemesi ile birlikte bu dönemde organın, depo organı haline geçişinde hızlanma olmuştur. Olgun yumruda dışarıdan itibaren birkaç sıralı ezik velamen, bir sıra ekzodermis, geniş bir depo parankiması (müsilaj hücreleri ve aralarında nişastalı hücreler) ve yer yer meristele görülmüştür.

Bitkiler, doğal antioksidan bileşiklerin başlıca kaynağını oluşturmaktadır. Fenolik bileşikler doğal antioksidanların en önemli gruplarını oluştururlar. Yapılan çalışmalarda, fenolik bileşiklerin; antialerjik, antienflamatuar, antidiyabetik, antimikrobiyal, antipatojenik, antiviral ve antitrombotik özellikleri ve kardiyovasküler hastalıklar, kanser, osteoporoz, diyabetes mellitus ve nörodejeneratif hastalıklarda koruyucu etkileri gösterilmiştir. Bu çalışmada Orchidaceae familyasından Ophrys mammosa subsp. mammosa türünden çiçeklenme aşamasında yumrular alınmış, bu yumrulardan LC-MS/MS ile fenolik bileşik analizi yapılmıştır. Analizi yapılan fenolik bileşikler gallik asit, protokateknik asit, 2,5-dihidroksibenzoik asit, kafeik asit, şırınga asidi (siringik), salisilik asit, klorojenik asit, kateşin, epikateşin, gibbarelik asit, 2-hidroksitranssinnamik asit, p-kumarik asit, verbascoside, rutin, sinapik asit, naringin, trans ferrulik asit, etil galat, indol-3-asetik asit, florizin, oleuropein, mirisetin, resveratrol, propil galat, kuersetin, absisik asit, lütolein, kaempferol, jasmonik asit, isorhamnetin, apigenin ve ellagik asittir. Yapılan analiz sonucunda Ophrys mammosa subsp. mammosa yumrularında gallik asit, protokateknik asit, 2,5-dihidroksibenzoik asit, klorojenik asit, p-kumarik asit, rutin, trans ferrulik asit, propil galat, kuersetin, lüteolein, kaempferol ve ısorhamnetin tespit edilmiştir

As a result of the detailed literature reviews, it has been determined that there are a limited number of studies on the stolon anatomy of orchids. In this study, Ophrys mammosa subsp. mammosa taxon by examining the stolon anatomy of the, it is aimed to contribute to the explanation of the tuber ontogeny of the genus. As material, microtome sections were taken from the embedding preparations with historesin kit in Ophrys mammosa subsp. mammosa stolons and stained with toluidin blue. From anatomical sections; It was determined that the vascular bundles in the middle part of the stolon are spaced and meristele distributed asymmetrically and the presence of statocyte cells in the lower part, the presence of a velamen in the outermost and several rows of exodermis under it, and an inner parenchymatic wide cortex. As a result; It has been emphasized that the anatomy of the orchid stolon is partially similar to the tuber that will develop in the future, and the importance of the findings in terms of tuber biology is discussed., Yapılan detaylı literatür incelemeleri neticesinde orkidelerin stolon anatomisine yönelik sınırlı sayıda çalışmanın bulunduğu tespit edilmiştir. Bu çalışma ile Türkiye’de yayılış gösteren Ophrys mammosa subsp. mammosa taksonunun stolon anatomisi incelenerek cinsin yumru ontogenisinin açıklanmasına katkı sağlanması amaçlanmıştır. Materyal olarak O. mammosa subsp. mammosa stolonlarında historesin kiti ile gömme preperatlardan mikrotom kesitleri alınmış ve toluidin blue ile boyanmıştır. Anatomik kesitlerden; stolonun orta kısmının boşluklu ve meristele düzeni iletim demetlerinin asimetrik olarak dağıldığı ve alt tarafında statosit hücrelerinin varlığı, en dışta bir sıra velamen ve altında birkaç sıralı ekzodermisin varlığı, içte parankimatik bir geniş korteksin olduğu tespit edilmiştir. Sonuç olarak; orkide stolon anatomisinin, ileride gelişecek olan yumruya kısmen benzediği vurgulanmış ve bulguların yumru biyolojisi açısından önemi tartışılmıştır.

In this study, the physio pathological effects of Aspergillus alliaceus (Aa, fungi, biocontrol agent) on Orobanche (parasitic plant) were investigated by hormone and phenolic substance tests. In experimental group, Orobanches were treated with the fungi, considering control group was fungus-free. Based on the hormonal tests, in the experimental group, salicylic acid (SA), jasmonic acid (JA), abscisic acid (ABA) and gibberellic acid (GA) levels significantly decreased, and only indole acetic acid (IAA) hormone levels were fairly higher than the control group. According to phenolic substance tests, it was found that only gallic acid, syringic acid and caffeic acid values significantly increased compared with control, and catechin and p-coumaric acid values were significantly lower. Consequently, it was determined that Aa pathogenesis (1) considerably reduces the effects of all defence hormones (JA, ABA, SA), (2) operates an inadequate defence based solely on the IAA hormone and several phenolic substances (gallic acid, syringic acid and caffeic acid), (3) and inevitably the fungi lead the Orobanche to a slow and continuous death. The results were evaluated in detail in the light of similar recent article and current literature in terms of biocontrol and pathology.

The aim of this study is to determine in detail the genotoxic effects of Olive Oil Wastewater (OOWW) on sunflower. For this reason, different concentrations of OOWW (1/1,1/10,1/100) were applied as irrigation water to sunflowers at different times (3-day, 5-day, 10-day). In the plants taken during these times, RAPD-based genomic template stability (GTS) assays and gene expression (transcriptomic) levels of different free radical scavenging enzyme genes (SOD, CAT, SOD2, GST, GPX, APX), protein repair/chaperoning genes (HSP26, HSP70, HSP83), N metabolism gene (GS) and apoptotic genes (BAX, BCL2, BCLXL, CYT-C, XIAP) were compared to the those of the control (OOWW-free) group. As a result; The GTS rates seemed to be fairly lower than the control and therefore the OOWW was likely to cause significant damage to the DNA's nucleotide and genomic structure, and the GTS value increased inversely proportional when the OOWW concentration was reduced from 1/1 to 1/10, and after a 10-day application, it seemed to be partly healing. In transcriptomic analysis; all OOWW experiments caused a free radical threat, and especially in 5-day OOWW applications, this raised significantly almost all expressions of antioxidants, protein repair, N metabolism, and apoptotic genes. So, the damages of 5-day OOWW treatments were found to be relatively more than those of 3-day treatments. Regarding 10-day transcriptomic data; a partial repair was found. Additionally, it was determined that the values of B, F, Al, Mn, Ni, Cr, As, Se, Cd, Pb and total polyphenols were high in OOWW. Our findings were also supported by plant images and various heavy metals' and OOWW polyphenols' toxicity results. Our results pointed to key findings in OOWW genotoxicology.

The aim of this study was to investigate the pathogenicity of Aspergillus alliaceus (Aa), a fungal biocontrol agent, on Orobanche by transcriptomic tests. For this reason, the expression of several genes was analysed comparatively in the experimental (fungi applied) and control (fungi-free) groups. After fungal infection, the expression of antioxidant enzyme genes SOD (Mn-superoxide dismutase), CAT (catalase), and SOD2 (Zn-superoxide dismutase) and the genes GS (Glutamine synthetase), HSP70 (heat-shock protein), and apoptosis [BCL2, BAX, caspase3 (CASP3)], which are related to protein metabolism, were investigated in the both groups. In gene expression studies, gene expression of SOD was almost 2.5 times higher in the experimental group than in the control group (fungus-free, intact), while the expression levels of other antioxidant genes (CAT and SOD2) were significantly reduced. While GS, which is the N and protein metabolism balancer, was found to be relatively low as compared to the control group, however, the level of HSP70 (protein folding, chaperoning) was quite higher than the control. Gene expressions of all apoptotic genes, BAX (=apoptosis suppressive gene), CASP3 and BCL (=apoptosis inducer genes), were significantly lower than control group. All the results found for the control group were determined to be statistically significant. It was concluded that during Aa pathogenesis, (1) the increased SOD value was associated with ROS (reactive oxygen species) threats, (2) the fungi disturbed protein synthesis metabolism, and finally, (3) the inhibited antioxidant and apoptosis-based pathways compared to the control group. These conclusions were discussed in the light of slow but constantly lethal effects of Aa comparatively to other pathogenic fungus, Fusarium.

The physiological effects of Fusarium oxysporum on in-root parasitic weed, Orobanche spp. (broomrape) with references to change in plant hormones and secondary plant constituents were investigated. The levels of IAA, GA, ABA and JA in the experimental group were significantly lower than those in the control group, while the level of SA was higher in the experimental group. In secondary metabolic studies, the quantities of various phenols were measured in the two groups and catechin, syringic acid and p-coumaric acid amounts were significantly higher in the experimental group than in the control group, unlike gallic acid which have a lower amount. Consequently, in the light of all data, it was concluded that Fusarium oxysporum (1) causes heavy hormonal disorder, (2) triggered only SA-mediated defense and (3) induced intensively accumulation of phenolic substances in orobanche. Fusarium oxysporum causes lethal physiological damage on Orobanche spp.

The pathogenic fungus Aspergillus alliaceus has been shown to have potential for the biocontrol of Orobanche spp. (broomrape), a root parasitic plant. The effectiveness of A. alliaceus in reducing Orobanche infection was analysed using pesta granules prepared with different food formulations. The results showed that pesta granules comprising of fungal mycelia/spore mixtures from liquid and solid culture, sclerotia and fungal mycelia reduced Orobanche infection to a greater extent in below ground conditions when applied early and at high doses before crop sowing. In addition, pesta granules eliminated the risk of broomrape contamination within a 0.2–0.3 cm diameter of the granules. The sclerotial pathogenicity of A. alliaceus was compared with those of other fungi reported in other studies. In addition, some morphological and histological studies on the fungal pathogenicity on broomrape plants after infection are presented. The present study reveals the potential of sclerotial A. alliaceus pesta granule ap...

This study aims to evaluate the toxic effects of Fusarium oxysporum on root parasitic weed, Orobanche spp. Comparative genetic and gene expression studies were conducted on uninfected and fungus-infected orobanches. In genetic studies, isolated total DNA was amplified by RAPD PCR. Fragment properties were analysed by GTS test. According to the results, the fragment properties of control and Fusarium infected (experimental) groups varied widely; and it has been observed that Fusarium has genotoxic effects on the DNA of orobanches. In gene expression studies, the expression levels of genes encoding enzymes or proteins were associated with ROS damage and toxic effects, therefore, gene expressions of Mn-superoxide dismutase (SOD), Zn-superoxide dismutase (=SOD2, mitochondrial), glutamine synthetase (GS), heat shock protein gene (HSP70), BAX, Caspase-3 and BCL2 were significantly higher in the experimental group. In the light of obtained data, it was concluded that F. oxysporum (1) caused heavy ROS damage in Orobanche (2) induced significant irrevocable genotoxic effects on the DNA of Orobanche, (3) degraded protein metabolism and synthesis, and finally (4) triggered apoptosis. The results of this study can be a ground for further research on reducing the toxic effects of Fusarium on agricultural products, so that advancements in bio-herbicide technology may provide a sustainable agricultural production.

In the present study, the morphological and histochemical properties of Himantoglossum robertianum (Orchidaceae) seeds were investigated. According to the morphological studies, the testa cells are dead and show reticulations. According to the histochemical tests, mature embryos were globular and covered with cuticular sheath in especially on the embryo parts of the seed. Furthermore, high amounts of soluble carbohydrates, lipid and protein bodies were found in mature embryos; but very little of starch was found. Consequently, the present results are important for both European-Mediterranean terrestrial orchids and present taxon, H. robertianum; thus they are discussed from the point of view of seed germination physiology and orchid embryology.

During extensive surveys in fields heavily infested by broomrape in the Trakya Region-Turkey, a different new fungus, Aspergillus alliaceus, was isolated from the infected broomrape. It is aimed to investigate whether or not it is really a pathogen for Orobanche. The fungi was exposed to a greenhouse environment in order to assess its pathogenicity and virulence against Orobanche cernua. In addition, infection tests on Orobanche seeds were also performed under laboratory conditions. The fungus was subjected using two different methods, exposure to a liquid culture with conidial solution and a sclerotial solid culture with fungal mycelia. Cytological studies were carried out at light, TEM and SEM levels. The results show that the sclerotial solid culture with fungal mycelia quickly caused necrosis and was more effective than the other type. It also greatly diminished attachments, tubercles, and caused the emergence of shoots and an increase in the total shoot number of Orobanche. In addition, both when the fungi was exposed to both soil and used to contaminate sunflower seeds, its pathogenicity was more effective. Consequently, it was determined that A. alliaceus was an effective potential biological control of broomrape throughout its life cycle from dormant seed to mature plant.

In the present study, it is aimed to investigate embryo and protoplast isolation from orchid seeds, because of know-how deficiency. As material Barlia robertiana seeds were subjected enzymatic process. Globular embryos with suspensor were fully removed by enzymatic maceration for between 100-180 min of incubation. The highest obtained embryo yields were found in Rapidase EX Color and Rapidase Vino Super, with degree of 89.2-90.3 × 103·g-1 MF and 86.4-91.3 × 103·g-1 MF, respectively. According to multicomponent maceration tests, a treatment comprising 150 min of the enzymatic reaction with 0.18% (E/S) at 50℃ was the most optimal for obtaining high embryo yields. Regarding protoplast isolation tests, enzyme combinations C (1% Cellulase “Onozuca” R-10 + 0.2% Macerozyme R-10 and 0.1% Driselase) and F (1% Cellulase “Onozuka” + 1% Macerozyme + 0.5% Driselase) after 15 hours produced the best results for protoplast isolation and were significantly different compared to other enzyme combinations. Greater protoplast yields were obtained using a rotatory system with protoplasts incubated in the dark. Also the present findings were discussed from the point of view of in vitro plant manipulation and orchid cultivation in cultural media and then their importance emphasized in molecular biological/genetical studies.

This article addresses the vegetative anatomy (leaves, stems, roots, root tubers and rhizomes) of 13 species of subfamily Orchidoideae (Orchidaceae), belonging to the genera Neottia Guettard, Cephalanthera L.C.M. Richard, Epipactis Zinn, Limodorum Boehmer, Spiranthes L.C.M. Richard, Platanthera L.C.M. Richard, Serapias L., Himantoglossum W.D. Koch and Anacamptis L.C.M. Richard, because anatomical studies have provided very useful criteria for orchid diagnosis. In the study three types of painting methods—Delafield’s hematoxylin and safranin, Alcian blue-periodic acid schiff, and alcoholic phloroglucinol + HCl—were employed, and identification tables were prepared. Anatomical results demonstrated the differences in the leaf anatomy of tuberous and rhizomatous orchids. In the stem anatomy, all the rhizomatous genera were found to be anatomically different, especially in regard to the collateral vascular bundles, the distribution of vascular bundles and xylem properties. In root anatomy, the central cylinder, pith, endodermis and/or pericycle properties are distinctive features in all studied taxa. For root tubers, velamen layering, wall outline mucilage cell patterns in ground tissue and arrangements of vascular arches can be used to label taxa. Regarding the rhizome anatomy of the studied taxa, vascular cylinder results in particular were very significant for the distinction of genera. Finally, we strongly emphasize the importance of this kind of detailed anatomical study to solve identification problems of orchid taxonomy.

The objective of this study was to investigate anther wall and pollen development in Ophrys mammosa. Primary sporogen tissue resembles longitudinal cells with divided archeosporal cells. Thereafter these primary sporogen tissue cells re-divide anticlinally and periclinally forming secondary sporogen tissue. Microsporogenesis was successive type. Microgametogenesis occurred at the distal poles of the microspores. In addition, dense starch accumulation was detected in the pollen. Pollinia and massulae are separated from each other by dead cells filled with callose, according to histochemical preparations. The anther wall was a four-layered “monocotyledon” type. There was ring-like wall thickening in the endothecium. The tapetum is of the glandular type. When these two developmental processes are compared, it is seen that the anther wall has become mature by the sporogen tissue phase and is composed of only epidermis and endothecium at the beginning of microgametogenesis.

Morphologies of pollen and seed surfaces were investigated with SEM technology for eight natural Ophrys taxa collected from southeastern Mediterranean countries and Turkey. The morphometric characteristics ofO. transhyrcana, O. sphegodes, O. epirotica, O. mammosa, O. pseudomammosa, O. oestrifera subsp.oestrifera, O. cornuta, andO. apifera were recorded and compared statistically. In general, the current species could be separated, in detail, according to those characteristics. These findings were also compared with those from recent studies, and were examined in terms of the present neighbouring taxa.

A new Onobrychis taxon was collected from regions of Turkey that reside in Europe and is described as Onobrychis mehmetchiquii Aybeke & Dane sp. nova belonging to sect. Onobrychis. Additionally, a description of the species along with illustrations and key features are given in contrast to O. alba and O. degenii which are the most similar to the new taxon.

The aim of this study is to assess the effects of Olive Oil Mill Wastewater (OOMW) application as irrigation water on in vitro pollen germination, focusing on total protein quantity. In test groups, pollen germination substances such as sucrose, H3BO3 and Ca(NO3)2 were added to different concentrations of OOMW and used as germination media. Regarding control group, the same substance melted into water instead of OOMW. As a result, in general, pollen germination percentage was decreased significantly in all OOMW concentrations than that of the control group, except 1/1000 concentrations. Similarly, total protein quantities declined linearly depending on decreasing OOMW concentrations, except 1/1 concentration which has 4-5 times the control value. Consequently, it was established that OOMW generally decreased pollen germination ratio and had carcinogenic effects on protein synthesis mechanism and must not be used as irrigation water without purification.

In this study, investigation of some anther structures in Sanguisorba minor Scop. subsp. muricata was aimed. Therefore, anthers were squashed by modified glycerine-gelatine method and additionally different histochemical stainings also were tested. The result illustrated that in different parts of anther, such as marginal, connection tissue, starch and protein granules accumulated. Furthermore, during maturation these deposits were detected in pollen grains also. In addition, a very interesting feature, balloon-like structures filled by proteins, were observed in anther cavity. In the light of these findings, their importance on pollen development was discussed.

A continuous aeropalynological survey of the atmosphere of Kirklareli was carried out from January 2002 to December 2003 by means of the gravimetric method using Durham apparatus. Weekly pollen grains in per cm(2) were calculated. During these 2 years, a total of 11,758 pollen grains were recorded. Pollen fall in the years 2002-2003 comprised grains belonging 46 taxa. Of these taxa, 26 belonged to arboreal and 20 taxa non-arboreal plants. In 2002, 6011 pollen grains and, in 2003, 5747 pollen grains were recorded. Total pollen grains consisted of 71.02% grains from arboreal plants, 28.93% grains from non-arboreal plants, and 0.05% unidentified pollen grains. Pollen from the following taxa we also found to be prevalent in the atmosphere of Kirklareli: Pinus spp., Cupressaceae/Taxaceae, Gramineae, Quercus spp., Chenopodiaceae/Amaranthaceae, Plantago spp., Platanus spp., Aesculus spp., Xanthium spp., Fagus spp., Robinia spp., Urticaceae, Betula spp., and Oleaceae, Artemisia spp. A total of 64.95% pollen grains appeared during April to June. The pollen calendar for the region presented in this paper may be useful for allergologists to make an exact diagnosis.

The anatomical properties of leaves, stems, roots and root tubers of 27 taxa belonging to three orchid genera (Ophrys, Orchis and Dactylorhiza) were investigated to determine their diagnostic characteristics and assess interspecific and intergeneric relationships by means of cladistic analyses. Anatomical sections were done using different staining methods, such as Sartur, Phloroglucin-HCl, Delafield's Hematoxylin-Safranin, and Alcian Blue-Periodic Acid-Schiff. Additionally, lengths of raphides (in all the organs), cuticle thickness, cell size and stomata dimensions (in the leaf epidermes), as well as dimensions of starch granules (only in root tubers) were measured and statistically compared. During root tuber investigations, for every taxon the types of meristele were outlined and diagrammed. All morphometric data except starch grain dimensions, are generally not significantly different between the investigated taxa. The anatomical sections showed the following basic anatomical characteristics: Leaf surface was glabrous, stomata were abaxially located and mostly anomocytic. Cuticle was mostly striate. The chlorenchyma was homogeneous. The stem had a uniseriate epidermis with several rows of thick- or thin-walled, sclerenchymatous (or collenchymatous) cortical parenchyms, a pericycle, collateral vascular bundles within a central ground tissue. Orchis purpurea lacks a pericycle. The roots generally had a uniseriate epidermis, an exodermis, a parenchymatous cortex with occasional inclusions of fungal pelotons and hyphae, an endodermis bearing casparian strips, pericycle, and vascular cylinders. Root tubers, from the outside to the centre, had orderly velamen, exodermis, and ground tissue, which were scattered with meristeles, mucilage cells and assimilatory cells bearing cruciate starch grains. Based on the anatomical results, tables were created based on the important primary (species-specific) and secondary (common-specific, used only in some taxa) descriptive characteristics. Some of the primary descriptive characters include the following: Leaf-epidermal wall properties (thin/thick), chlorenchyma cell shape, chloroplast distribution, presence or absence of large lacunae; stem-presence/absence of pericycle, cortical thickness; root-cell shape of exodermis, presence/absence of tilosome passage cells, shapes of endodermal and/or pericycle cells, structures and distributions of archs; root tuber forms of starch grains and mucilage cells and meristele types. In general, leaf chlorenchyma and stem pericycle were found to be thicker and mucilage-water storage cells were more frequent in the Orchis and Dactylorhiza taxa. Cladistically analysed anatomical data support the hypothesis that Ophrys and Dactylorhiza are monophyletic, but contrast the triphyletic states of Orchis, which is very likely to be the reason for uncovering the other Orchis taxa. Finally, we strongly emphasize the importance of this kind of detailed anatomical study to solve identification problems of orchid taxonomy, which have wrongly been disregarded in recent works. (C) 2009 Elsevier GmbH. All rights reserved.

Toxic effects of the wastewater were investigated ultrastructurally in root tips of Triticum aestivum. As a result, wall and nuclear degradations, disruptions in all cytoplasmic membranes, irregular nucleus shapes and cellular organization defects were densely detected. Besides, germination ratio, total protein contents, DNA contents and root-shoot growth were found to be decreased significantly when compared to the control group. Results were compared with those of recent studies regarding excessive Na+, Fe+2, P, polyphenols and acidic pH toxicity.