* National Institute of Research and Development for Biological Sciences Bucharest, “Stejarul” Biological Research Centre, Alexandru cel Bun, 6, 610004, Piatra Neamt – Romania
** “Alexandru Ioan Cuza” University of Iasi, Faculty of Biology, Bd. Carol I 20A, 700505, Iasi – Romania
*** Zürich University of Applied Sciences, Institute of Biotechnology, Einsiedlerstrasse 31, 8820 Wädenswil – Switzerland

Abstract: Arnica montana L. is an important plant bioresource, being traditionally used as medicinal plant, for which the scientific and economic interest remains at a high level. The aim of our study was to evaluate the bioproductivity for Arnicae folium et caulis in terms of biomass and content in biological active compounds originating from the wild populations and in vitro experimental cultures – under controlled environment. In situ biomass production registered variations, probably due to the different environmental features and also to the management strategies for each site. The development of in vitro cultures had as purpose the evaluation of the biomass production and production of plant material for the phytochemical screening. The Arnicae folium et caulis samples harvested from the wild populations have a higher content in phenolic acids than the Arnicae flos from the same area, but the total content in sesquiterpen-lactones was significantly lower. In contrast, the samples originating from the in vitro cultures had a low content in phenolic compounds and a high content in sesquiterpen-lactones, comparable with Arnicae flos.

Key words: A. montana, biomass, phenolic acids, flavonoids, sescviterpen-lactones, HPTLC, HPLC

* Faculty of Agriculture and Graduate School of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795, Japan

Abstract: Plant growth regulators (PGRs) were used to induce callus in 15 carnation (Dianthus caryophyllus L.; Caryophyllaceae) cultivars: Orange Sherbert, Avalanche, Magenta, La France, Stripe Red, Marie, Concerto PVP, Snap, Lucky Pierot, Cinnamon Tea, White Love, Siberia, Magesta, Spark Bruno, and Honono no Estejo. Seeds were initially sown on autoclaved moistened filter paper and internodes of surface-sterilized seedlings were used as explants. Most callus was induced in the presence of 0.5 mg/L a-naphthaleneacetic acid used together with 1 mg/L 6-benzyladenine or 1 mg/L 2,4-dichlorophenoxyacetic acid on basal Murashige and Skoog medium. Callus is not a desirable method to clonally propagate important germplasm but can serve as one possible way of deriving periclinal mutants as a result of somaclonal variation.

Key words: carnation, internode, plant growth regulator, thin cell layer

* Biotechnology and Molecular Biology Laboratory, Forest Genetics and Tree Breeding Division, Arid Forest Research Institute, New Pali Road, Jodhpur 342005 – India
** Indian Council of Forestry Research and Education, P.O. New Forest, Dehradun 248006 – India

Abstract: Hardwickia binata Roxb. is a leguminous tree of great economic importance. Yet a protocol for its in vitro propagation that is capable of taking the regenerated plants from lab to land does not exist. The plant is quite recalcitrant and has poor natural regeneration. Macropropagation techniques are also not standardized. Problem of leaching of phenolic compounds in culture condition, premature leaf fall during multiplication phase and callusing during rooting were sever problems encountered during the course of development of the protocol. These problems were addressed using various treatments and a working protocol for in vitro propagation of H. binata from axillary bud of the seedling nodal segments was perfected. The plantlets were hardened through a systematic two stage hardening procedure and were eventually transferred to experimental field. Here we report the development of an in vitro propagation protocol for H. binata, leading to successful establishment of plants thus obtained, in open field condition.

Key words: micropropagation, shoot multiplication, rooting, acclimatization, RAPD

EMBRYOLOGICAL CHARACTERS TO STUDY THE JUSTICIA-RUNGIA COMPLEX (ACANTHACEAE)
Nitin M. LABHANE*, Nitin M. DONGARWAR*

* Department of Botany, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur 440033 – India

Abstract: Family Acanthaceae is characterized by very diverse plants whose taxonomic position is debated. Therefore, study of various macroscopic and microscopic characters had been used for studying the relatedness of the various taxa. Embryological characters are considered as one of the most stable characters. Justicia–Rungia Complex is one of the intriguing complexes present in family Acanthaceae. Justicia is basically a Linnaean Genus which included Rungia also. But later Rungia was separated from Justicia by Nees. Another offshoot of Justicia has been the formation of new genus Rostellularia which is also the creation of Nees. Due to this the Rungia is sometime confused as Justicia and some time it is considered as Rostellularia. In the present investigation embryological data is used to study the Justicia–Rungia Complex. The plants studied in present investigation are Justicia procumbens, Rungia repens, Haplanthus verticillata and Blepharis repens. The embryological study using UPGMA clearly indicates a very close similarity between Justicia procumbens and Rungia repens.

Key words: Embryology, Acanthaceae, Justicia-Rungia Complex, UPGMA

ANATOMY AND ONTOGENY OF ASTRAGALUS REMOTIJUGUS BOISS. & HOHEN. SEED
Mozhgan FARHANGISABET*, Ahmad MAJD**, Taher NEJADSATARI*, Ali MAZOOJI***, Aliasghar MAASOMI ****

* Science and Research Branch Islamic Azad University, Department of Biology, Tehran – Iran
** North Tehran Branch Islamic Azad University, Department of Biology, Tehran – Iran
*** Roodehen Branch Islamic Azad University, Department of Biology, Roodehen – Iran
**** Research Institute of Forests and Rangelands, Department of Botany, Tehran – Iran

Abstract: The aim of this study was to describe the anatomy and ontogeny of Astragalus remotijugus Boiss. & Hohen. seed using the usual techniques. The ovules are campilotropous, crassinucelate, and bitegmic. The nucellus cells disappear during an early stage of development. The following processes occur during integument development: anticlinal divisions and formation of palisade cells in the exotesta; predominantly periclinal divisions and cell expansion in the mesotesta; and endotesta differentiates in to an endothelium layer with thick and cubic in shape cells. The primary endosperm nucleus divides before the zygote nucleus, forming a nuclear endosperm. Endosperm cellularization begins when the embryo has developed the late globular stage. The embryological characters of A. remotijugus Boiss. & Hohen. are compared with other species of Fabaceae and those of other species of Astragalus. The remarked characteristic of the embryo was presence large suspensor with six columns of cells, and its large haustorial cells. Another difference in the development of A. remotijugus Boiss. & Hohen. seed was presence additional embryo at the globular stage that probably was developed from suspensor cells.

Key words: embryogenesis, Fabaceae, polyembryony, seed coat, suspensor

ULTRASTRUCTURAL ASPECTS OF PROGRAMMED CELL DEATH IN THE EXOCARP OIL GLANDS OF MANDARIN (CITRUS DELICIOSA TEN.)
Artemios Michael BOSABALIDIS*

* Department of Botany, School of Biology, Aristotle University, Thessaloniki 54124 – Greece

Abstract: In the exocarp of mandarin fruit (Citrus deliciosa Ten.), numerous globular/ovoid oil glands occur. In the centre of each gland, an essential oil-accumulating cavity is formed by a process of cell lysis. This process is induced by PCD which becomes ultrastructurally evident by the presence of a large number of fragmented ER-elements with a dark content. They appear only at the stage of PCD initiation and they disappear afterwards. ER-elements are scattered over the entire cytoplasmic area and do not locally aggregate or associate with other cell organelles and particularly the vacuoles. TEM observations favour the interpretation that ER involves in PCD of oil gland cells by releasing hydrolytic enzymes directly to the cytosol.

Key words: Citrus deliciosa Ten., endoplasmic reticulum (ER), hydrolytic enzymes, oil cavities, programmed cell death (PCD)

* Department of Botany, Rashtrasant Tukadoji Maharaj Nagpur University, Nagpur 440033 – India

Abstract: The study of morphology of embryo is very significant considering the fact that the embryo represents the important step in the determination of the viability of the seed. Ruminate endosperm has been reported in about 58 families of angiosperms. The rumination caused by the activity of the seed coat or by the endosperm itself is quite recurrent in angiosperm. Ruminate endosperm due to seed coat is reported from the family Acanthaceae in Andrographis paniculata. The rumination of endosperm is also considered as phylogenetically important. Rumination of endosperm is very common, however very little is known about rumination in embryo. The present papers reports the de novo development of ruminate embryo in Blepharis repens. The development of ruminate embryo is seen as an adaptation to ensure proper aeration and optimum germination for survival of the species.

Key words: rumination, seed, ruminate embryo, endosperm

* Jai Narain Vyas University, Laboratory of Plant Ecology, Department of Botany, Jodhpur 342 033 – India

Abstract: The present article deals with seed germination behaviour of Commiphora wightii (Guggal) by using various concentrations (0, 5, 10, 15 & 20 mg L^-1) of different nitrate solutions such as NH₄NO₃, Co(NO₃)₂, Ca(NO₃)₂ and KNO₃ under nursery conditions. At present, plant is considered under data deficient category. The seeds were collected from four different sites of three districts in western Rajasthan, viz. Jodhpur, Jaisalmer and Barmer. Results revealed that seeds presoaked in Ca(NO₃)₂ and KNO₃ were found to be most suitable for germination under nursery conditions as compared to others. The best results for germination initiation, germination duration (days), germination percentage, peak value (PV), root and shoot lengths, collar diameter and biomass yield were also observed maximum in these two nitrate pretreatments.

Key words: Commiphora wightii, nitrate solutions, germination initiation day, peak value (PV), data deficient, medicinal plant

* Department of Biology, Faculty of Science, University of Guilan, Rasht – Iran

Abstract: High salinity causes ion poisoning and subsequent oxidative stress. During oxidative ion poisoning, non-enzymatic defense systems such as carotenoids, phenols and flavonoids are activated. In this study, the effects of various concentrations of chloride in the irrigation water (10, 20, 40 and 80 mg/l) on carotenoids, phenols, flavonoids and proline content in three varieties of tobacco plant (Virginia, Kentucky and Cooker) were investigated. Malondialdehyde content was also measured, as a lipid peroxidation index. The highest level of ß-carotene was observed in Virginia at 10 mg/l chloride and the lowest quantity was seen in Kentucky at 20 mg/l chloride. Kentucky had the highest and lowest levels of flavonoids at 80 and 40 mg/l concentrations of chloride respectively. Highest phenol content was observed in the presence of 10 mg/l chloride in Virginia. Maximum and minimum MDA concentrations were observed respectively in 20 and 80 mg/l concentration of chloride in Cooker cultivar. Raising chloride concentrations in irrigation water caused a substantial decrease in proline content only in the Kentucky variety.

Key words: Oxidative stress, ß-carotene, chloride, flavonoids, tobacco, total phenol

* Kharazmi University, Faculty of Biological Science, Department of Plant Biology, Tehran – Iran

Abstract:Tehran pine (Pinus eldarica Medw.) belongs to Pinaceae family with significant economic and ecological benefits. To gain further insight into anatomical-developmental structure of P. eldarica, both the vegetative and generative meristematic tissues and microsporogenesis were been studied during certain stages of development. To do this, meristematic tissues and male cones were initially fixed in FAA solution (37% formaldehyde, 96% ethanol and glacial acetic acid with a 2:7:1 ratio, respectively). They were then embedded in paraffin and sectioned using a rotary microtome. Prior to visualization and photography under a camera-equipped light microscope, they were stained with hematoxylin-eosin (Zeiss model). Our results revealed the vegetative meristem of P. eldarica to be in the Cryptomerya-Abies category. The results also indicated it is a protuberant (dome-like) type containing four regions. The meiosis occurs before the winter dormancy and continues through the winter. The pollen is shed at the four-cell stage of development.

Key words: cone development, generative meristem, microsporogenesis, pollen grain, Pinus eldarica, Tehran pine

PRODUCTIVITY AND CYTOGENETIC STABILITY OF PROTOCORM-LIKE BODIES OF HYBRID CYMBIDIUM CRYOPRESERVED BY ENCAPSULATION-DEHYDRATION AND VITRIFICATION
Jaime A. TEIXEIRA DA SILVA*

* Faculty of Agriculture and Graduate School of Agriculture, Kagawa University, Miki-cho, Kagawa, 761-0795 – Japan

Abstract: Transformed and untransformed protocorm-like bodies (PLBs) of hybrid Cymbidium Twilight Moon ‘Day Light’ were cryopreserved using two methods (encapsulation-dehydration and vitrification) to form new PLBs (neo-PLBs) on Teixeira Cymbidium (TC) medium. The organogenic response was quantified and genetic stability was assessed by flow cytometry. Intact PLBs produced significantly more neo-PLBs and fresh weight of neo-PLBs than half-PLBs or PLB longitudinal thin cell layers (lTCLs), for two vitrification protocols and for an encapsulation-dehydration method. The addition of 2% sucrose significantly improved the number of neo-PLBs than half-PLBs or PLB lTCLs but any other concentration of sucrose, and all other osmotic agents (mannose, PEG-6000, DMSO) at any concentration resulted in significantly worse neo-PLB formation. However, the length of exposure to sucrose did not significantly affect the number or fresh weight of neo-PLBs. The desiccation (air current or silica gel) method applied negatively affected the number and fresh weight of neo-PLBs. The length of the cryopreservation period negatively impacted the number of neo-PLBs and fresh weight of transformed or untransformed neo-PLBs. Most severe negative effects were registered after one year of cryopreservation. Cryopreserved neo-PLBs showed high levels of endopolyploidy (8C-64C) relative to non-cryopreserved PLBs.

Key words: alginate bead, orchids, PLB, thin cell layers, Teixeira Cymbidium (TC) medium, transgenic

* CSIR-Crops Research Institute, P.O. Box 3785, Kumasi – Ghana, West Africa
** Universität Hohenheim, Emil-Wolff-Str. 25, D – 70593, Stuttgart – Germany

Abstract: One major constraint to plantain production has been inadequate healthy planting materials at the time of planting. Several technologies for multiplying healthy planting materials exist but could not meet farmers’ demand. A study was conducted to assess the performance of various landraces plantain to plants issus de bourgeons secondaires (PIBS) technique. Five cultivars of Musa sapientum (Apantu (False Horn), Asamienu (True Horn), Oniaba (intermediate French plantain) and FHIA-21 (tetraploid hybrid plantain) were tested to determine their response to the PIBS technique. Sword suckers of each cultivar with weight of between 0.2-0.5 kg were prepared and buried in fine sawdust in a humidity chamber built using transparent polyethylene sheets. Results at harvest showed that removal of rooted sprouts started three weeks after planting and every week thereafter for eight weeks. The intermediate French plantain cultivar (Oniaba) produced the least average number (about 20) of healthy planting. Apantu (False Horn) produced an average of about 75 healthy planting materials. The hybrid FHIA-21 on the other hand generated an average of about 85 healthy planting materials. Asamienu (True Horn) produced the highest healthy seedlings of about 90 healthy planting materials. The results revealed that the leaf scar carries a primary bud at the intersection of each leaf sheath and several eyes along the entire length of the leaf sheath which could not have developed into suckers. However, with this technique the eyes could be activated to sprout as healthy planting materials. The technique proved as an efficient method of multiplying healthy planting materials for plantain and could thus be recommended for adoption not only by peasant farmers but also to others who could become commercial seed producers. But there will be a need for certification guidelines for seed growing systems.

Key words: Musa, plantain, macro-propagation, planting material, PIBS

* Actavis, 11 Ion Mihalache Blvd., 011171, Bucuresti – Romania
** Department of Botany, Institute of Biology, Eszterházy Károly College, Eger – Hungary

Abstract: Draba simonkaiana Jáv. is a critically endangered, Southern Carpathian endemic species, distributed only in Parâng, Retezat and Cozia Mountains. As an European endemic, and restricted to a single European country, D. simonkaiana is treated as a “species of European concern”.
Although of great international significance, the distribution of Draba simonkaiana is slightly known, without any georeferenced records on GBIF.
Based on field studies, analyses of herbarium material and literature data, the authors managed to record the occurrence of D. simonkaiana in Southern Carpathians and determined the threatened status according to criteria and categories of IUCN. Unfortunately, the species has not been found in locus classicus (Badea Hill) but the authors have discovered two new localities (in Parâng Mountains) of this important endemic species.

Key words: Draba simonkaiana Jáv., endemic species, Southern Carpathians, Romania, threatened species, Red Book, chorology

* Actavis, 11 Ion Mihalache Blvd., 011171, Bucuresti – Romania
** “Muntii Maramuresului” Nature Park, 22 Decembrie Str. 20, 435700, Viseu de Sus – Romania
*** “Alexandru Ioan Cuza” University of Iasi, Faculty of Biology, Carol I Blvd. no. 20A, 700505, Iasi – Romania

Abstract: Maramuresului Mountains Nature Park is widely known as one of the last wilderness areas in Europe and also represents a real oasis for naturalists eager to explore the flora and fauna of this special land not very researched.
During a botanical trip in the area of Farcau Peak (on 19 July 2014) the authors of this paper found three very rare species (all 3 threatened, included in Romanian Red Book of Vascular Plants): Trifolium lupinaster L., Heracleum carpaticum Porcius and Ranunculus thora L.
Based on field studies, analyses of herbarium material and literature data, the authors managed to record the occurrence of Trifolium lupinaster, Heracleum carpaticum and Ranunculus thora in the Romanian Carpathians and determined the threatened status of species according to criteria and categories of IUCN.

Key words: chorology, Eastern Carpathians, endemic species, “Maramuresului Mountains” Nature Park, Red Book, threatened species

* “Alexandru Ioan Cuza” University of Iasi, “Anastasie Fatu” Botanical Garden, Dumbrava Rosie, 7-9, Iasi – Romania
** “Alexandru Ioan Cuza” University of Iasi – Faculty of Biology, Carol I, 20A, 700505, Iasi – Romania

Abstract: Dead wood fungi have a major importance for forests biodiversity as they produce wood degradation in forest habitats. In this paper are presented some aspects related to the diversity of dead wood fungi in tree deciduous forest types from tree natural reserves (Crujana, Dragomirna and Zamostea) from Suceava County and the effect of some ecological factors (host tree, diameter and decomposition degree of the dead wood and some microclimatic characteristics of sites) on their occurrence and diversity. Investigations carried out in 2013 resulted in the identification of 44 lignicolous fungi species. Analysis of similarities between lingnicolous fungi species from the investigated natural reserves (by hierarchical clustering) shows a separation of three fungi groups, depending on the host-trees species. The effect of the tree host species was highlighted also by detrended correspondence analysis, which, in addition presented the existence of an altitudinal gradient and a weaker effect of site conditions (slope and aspect) and microclimatic variables (solar radiation) on dead wood fungi occurrence. The effect of diameter and decomposition degree of fallen trunks and branches on dead wood fungi species was investigated using the redundancy analysis showing that wood debris with large surfaces are more easily colonized by the fungi species developing large sporocarps compared to small branches with low diameters colonized only by few or a single fungus species.

Key words: forest, lignicolous fungi, wood debris, Detrended Correspondence Analysis (DCA), Redundancy Analysis (RDA)

* University of Agricultural Sciences and Veterinary Medicine Iasi, Faculty of Agriculture, 3, Mihail Sadoveanu Alley, 700490, Iasi – Romania
** Botanical Garden of the University “Alexandru Ioan Cuza”, 7-9, Dumbrava Rosie Str., 700487, Iasi – Romania

Abstract: This article contains a brief overview of the main aspects of the botanical opera of the Professor Valeriu Zanoschi, as well as a list of scientific papers and books published by him during the 38 years of activity in the service of the Romanian botany.