Phalaenopsis Amabilis Research Articles

Clonal Fidelity of Micro-propagated Phalaenopsis Plantlets Based on Assessment Using Eighteen Ph-Pto SNAP Marker Loci

Phalaenopsis amabilis is an Indonesia native orchid species having large, white flowers with yellow labellum coloration. This studies aimed to develop Phal. amabilis micropropagation methods and evaluate the regenerated plantlet fidelity. Media supplemented with Thidiazuron (TDZ) and Polyvinylpyrrolidone (PVP) and medium pH adjustment effects to induce protocorm-like bodies (PLBs) from leaf explants and proliferate secondary PLBs were investigated. Clonal fidelity among regenerated plantlets was evaluated using eighteen SNAP marker loci. The results showed that the ½ MS medium supplemented with 3 mg L-1 TDZ and 0.5 g L-1 PVP was the best for PLB induction while the ½ MS medium supplemented with 0.5 mg L-1 TDZ was the best for PLB proliferation. For PLB induction, the media pH was adjusted into pH=7 for efficient PLB regeneration. Based on the assessment using 18 SNAP marker loci, four variant alleles in three loci (11.8%) out of a total 34 plantlets were detected. The mutation frequency at the evaluated SNAP marker loci was 2.5 x 103 (0.25%). Changes in SNP alleles may not always result in phenotype changes and allele variant occurrences may not affect phenotype fidelity of micro-propagated Phal. amabilis plantlets. Therefore, further studies about the phenotype fidelity among plantlets are necessary.

PHENOTYPIC AND MOLECULAR CHARACTERIZATION OF MULTISHOOTS DEVELOPMENT IN TRANSGENIC Phalaenopsis amabilis (L.) BLUME HARBORING 35S::KNAT1 (KNOTTED-LIKE Arabidopsis thaliana 1)

Phalaenopsis amabilis (L.) Blume is one of Indonesian natural orchid which has an aesthetic flower and possesses high economic value. The low multiplication rate and long periods of life cycle are the main obstacles to conventionally propagate this orchid. The aims of this research were to analyze the stability of transgenic plant P. amabilis harboring 35S::KNAT1 based on morpho-genomic characterization. KNAT1 gene is reported as a gene that involved in the shoot formation, and it  had been successfully introduced into Phalaenopsis amabilis (L.) Blume genome. After seven times regeneration, the confirmation of the transgene existence in the genom is needed to ensure whether the plant could consistently maintain the transgene in its genome and to characterize the shoot development. The experiment was carried out in 3 steps: 1) Co-integration analysis of 35S::KNAT1 into P. amabilis genom; 2) Phenotypic analysis on the multiplication rate, morphological variation and venation pattern; and 3) Protein profile analysis of transgenic plants. The results showed that the survival rate of putative transgenic was 58.7% on NP0 medium and 62.5% on NP SIM medium. PCR analysis confirmed that 82.5% transgenic growth on NP0 and 93.33% on NP SIM contained DNA fragment of KNAT1 gene, NPTII gene and trnL-F intergenic spacer, indicating that those plants are positive transgenic. The 35S::KNAT1 transgenes and phytohormone were independently involved in multishoots formation of P. amabilis transgenic plants. The phenotypic of plantlets were classified into six main criteria, i.e. normal shape, lobed leaves, rosette, elongated stem, cup shoot and widened leaves. The normal type was the most abundant type of variation (± 29%) in both medium. Protein profile showed that all transgenic plants produced 45,8 kDa protein and that was equivalent with molecular weight of KNAT1 protein. Taken together, all those data indicated that 35S::KNAT1 transgene were consistently integrated into the transgenic plant genome.

Aplikasi silika dan NAA terhadap pertumbuhan Anggrek Bulan (Phalaenopsis amabilis l.) pada tahap aklimatisasi

The purpose of the research was to study the influence silica and concentration of NAA for moon orchid (Phalaenopsis amabilis L.) growth in acclimatization. The experiment was assigned in completely randomized factorial design with the first factor was silica (non silika and with silica) and the second factor was NAA concentrations (0, 50, 100, and 150 ppm). Each treatments was repeated three times. Parameters measured were the percentage of plant life, number of leaves, length of leaf, width of leaf, number of roots, length of root and fresh weight of plants. Data were subjected to ANOVA and followed by Duncan Multiple Range Test. The result showed that aplication silica increased fresh weight of plants. Application of NAA concentrations (50, 100, and 150 ppm) decreased percentage of plant life, leaght of leaf, and fresh weight of plants. Interaction of silica and NAA concentrations showed did not significantly effect. Keyword : Moon orchid, silica, NAA, aclimatization

The Influence of Thidiazuron on Direct Somatic Embryo Formation from Various Types of Explant in Phalaenopsis amabilis (L.) Blume Orchid

Phalaenopsis amabilis is an important national flower of Indonesia as a parent for orchid breeding, so that needs a good strategy to produce high number of plants. The objective of this research is to analyze the use of thidiazuron (TDZ) for producing high number of plantlets, through directly induction of somatic embryos (SEs) from various explants. The method was used 20 each of protocorms, leaves, stems and roots as explants. The explants were dissected transversely, then put on various culture media: New Phalaenopsis (NP) and NP + (1, 2, 3) mgL−1 TDZ. Cultures were maintained at 25°C with continous white light. The formation of SEs was observed every week for 8 weeks. The results showed that SEs formation increased inline with the addition of TDZ concentration to the NP medium, for both velocity and amount of SEs formation. In NP0, SEs were formed at (26.07 ± 0.73) days after inoculation of protocorm, whereas on NP + (1, 2, and 3 mgL−1) TDZ, SEs were formed at (17.85 ± 0.67) days, (15 ± 0.64) days, and (11 ± 0.64) days, respectively. All types of explants formed SEs on NP + TDZ (1–3 mgL−1), whereas only 14 of 20 protocorms produced SEs (70%), and 8 of 20 stems formed SEs (40%) in NP0. In roots, SEs was formed on NP + 2 mgL−1 TDZ and NP + 3 mgL−1 TDZ. For stems, the highest amount of SEs (28.25 ± 1.07) was reached on NP + 3 mgL−1 TDZ, followed by protocorm (23.30 ± 1.13) SEs and roots (8.25 ± 0.68) SEs. In contrast, in NP0, the amount of SEs was very low (1.25 ± 0.46) from stem and (1.50 ± 0.65) from protocorms, there was no evidence of SEs formation in the leaves and roots.

Uji Ketahanan Planlet Anggrek Bulan (Phalaenopsis amabilis (L.) Bl.) Hasil Seleksi dengan Asam Salisilat Terhadap Fusarium oxysporum Secara In Vitro

Phalaenopsis amabilis cultivated have many constraints such as the appearance of fusarium wilt disease caused by Fusarium oxysporum. Plantlet P.amabilis which resistant to F. Oxysporum was selected in the solid Vacin and Went (VW) medium was added with salicylic acid at concentrations of 65 ppm, 75 ppm, and 85 ppm, compared with controls (0 ppm). The goals of the research were to study and determine of: 1) The SA concentration of planlet P. amabilis selection tolerant; 2) The proper concentration of AS during in vitro selection for suppressing the Fo. The research was carried out in December 2015 to February 2016 in the Botany Laboratory (in vitro research room), Departement of Biology, Faculty of MIPA, Lampung University. The result showed that: 1) The SA tolerant concentration for plantlet selection with P. amabilis was between 65 ppm - 85 ppm. 2) The 85 ppm of SA was effective for suppressing the Fo compared to 65 ppm and 75 ppm. Keywords: Phalaenopsis amabilis, salicylic acid, Fusarium oxysporum, In vitro, Resistance.

Pemberdayaan Kelompok Lanjut Usia melalui Pengenalan Teknologi Pembuatan Pupuk Organik Cair (POC) sebagai Upaya Bentuk Kepedulian terhadap Lingkungan

In order to avoid post power syndrome, the elderly who have a hobby of farming / gardening can be empowered through the introduction of LOF (liquid organic fertilizer) making technology and its application. The aim is for the elderly to understand the technology of making organic fertilizers that are lightweight, useful and entertaining as well as for them to feel contribute in maintaining the environment. The training method applied is a combination of interactive lecture and practice methods. Based on these methods, have succeeded in growing their creativity in making LOF and feel comforted on the results obtained. Successful LOF products contained nitrogen, phosphorus and potassium (NPK) of 146,701 mg / L, 0.741 mg / L, and 0.035 mg / L, respectively, and magnesium (Mg) and calcium (Ca) of 86.332 mg / L and 1.970 mg / L. The application of LOF products has been done on the orchid plants (Phalaenopsis amabilis) with satisfactory results. Thus, the mastery of LOF making technology and its application will reduce the dependence of chemical fertilizer use and replace it with homemade organic fertilizer so it can contribute in maintaining health and environmental sustainability.

Isolation of Flower Development Regulator Gene SEPALLATA 1 in Phalaenopsis amabilis (L.) Blume

<p class="Els-Abstract-text"><em>Phalaenopsis amabilis </em>(L.) Blume<em> </em>is an indigenous orchid species in Indonesia. This orchid has a white large flower. The large flower is caused by the existence of gene that has an important role in flower development. One of the genes is <em>SEPALLATA 1. </em>This gene is a member of superfamily MADS-Box gene. <em>SEPALLATA 1 </em>gene is a marker of primordial flower organ. This study aimed to isolate <em>SEPALLATA1</em><em> </em>gene from <em>Phalaenopsis amabilis </em>(L.) Blume by PCR using forward primer 5’-GCT-GGA-GCG-GAT-CGA-GAA-CA-3’and reverse primer 5’-TCA-TGC-AAG-CCA-ACC-AGG-TG-3’<em>. </em>This study successfully amplified 691 bp lengths of <em>SEPPALATA1</em> fragment, lacking 20 bp upstream which consist its start codon.</p><p> </p><div><p class="Els-keywords"><strong>Keywords:</strong> Flower development regulation, <em>Phalaenopsis amabilis</em> (L.) Blume, SEPALLATA 1 gene.</p></div>

Deteksi Odontoglossum ringspot virus pada Anggrek Asli Koleksi Kebun Raya di Indonesia

Native orchid is one of Indonesian natural resources which play important role as parental materials in breeding program. Virus infection is one of the limiting factors in the cultivation of orchid. The purpose of this study was to detect Odontoglossum ringspot virus (ORSV) from native orchid. Symptomatic orchids were collected from 5 botanical gardens, i.e. Bogor, Cibodas, Purwodadi, Balikpapan, and Enrekang Botanical Gardens. Detection and identification was conducted by serological method using ORSV specific antisera, followed by RT-PCR and DNA sequencing. The serological test showed that 5 samples gave positive reaction against ORSV antiserum, i.e. Phalaenopsis amboinensis (KRB2) and Phalaenopsis amabilis (KRB12) from Bogor Botanical Garden, Phalaenopsis amabilis (KRP18) and Dendrobium salacence (KRP20) from Purwodadi Botanical Garden, dan Phalaenopsis modesta J. J. Sm. (KRBp5) from Balikpapan Botanical Garden. RT-PCR of the 5 samples using specific primer of ORSV coat protein gene was successfully amplified fragment DNA with size ± 474 bp. Homology analysis of those 5 ORSV isolates showed the highest index similiarity of 99.8% with corresponding sequences from 14 other ORSV isolates. Phylogenetic analysis indicated that ORSV KRB2 and KRP18 isolates was clustered in a separate group far from ORSV isolates in other countries. This is the first report of ORSV infection on native orchids collection from 5 botanical gardens in Indonesia.

PEMANFAATAN LIMBAH BUAH PISANG DAN AIR KELAPA SEBAGAI BAHAN MEDIA KULTUR JARINGAN ANGGREK BULAN (Phalaenopsis amabilis) TIPE 229

Coconut water and rotten banana fruits are commonly found in traditional markets as organic wastes. One way to overcome the problems caused by these organic wastes is to convert these unuseful matter into an important and economically useful matter by using them as components of tissue culture media. One important commodity that is usually propagated by tissue culture is Phalaenopsis orchid type 229 (Phalaenopsis amabilis). Therefore, it would be more benefit to substitute the expensive chemicals with organic wastes such as coconut water and banana puree.In this experiment, addition of coconut water and banana pure to the minimum media containing commercial fertilizer red Polyhyponex, sucrose and commercial agar did not show any inhibition of Phalaenopsis orchid plantlet growth. This probably caused by sufficient macro and micro nutrients provided by those organic matter and Polyhyponexfertilizer. Moreover, addition of 100 mL/L of coconut water and 100 mg/L banana puree gave the optimum leaf and adventitious shoot formation. On the other hand, addition of 150 mL/L coconut water gave the optimum height and root formation. In this case,growing Phalaenopsis orchid plantlet should be done in 2 subculture period. The first subculture is to obtain maximum amount of leaf and shhot formation while the second subculture is to obtain optimum height and root formation.Key words : Waste, coconut water, banana puree, tissue culture, Phalaenopsis orchi

Bawean Island: The Potential for Ecotourism and Local Knowledge on Plant Diversity Supporting Ecotourism

Bawean Island has attractive biodiversity and natures that are potential for development. The present study was aimed to investigate (i) the potential of Bawean Island’s natural resources for ecotourism (ii) local knowledge of Bawean people on plant diversity and uses, and (iii) the potential of plants for commodities supporting ecotourism development. The potential of Bawean Island for ecotourism was examined based on data of attractive natural environments, while the local knowledge was obtained by interviewing Bawean people and observation of the daily practical use of plants. Various plants were analysed for their potential as commodities suppporting ecotourism. The results showed that Bawean Island has high potential resources for ecotourism. It is ranging from beaches, mountains, rivers, plant diversity, and endemic animals. The local knowledge showed that Bawean has plant diversity with various potential uses. There were 81 plant species traditionally used by Bawean people, including plants for foods (40 species), ornamental plants (15 species), timbers for housings and buildings (9 species), medicines (8 species), natural dyes (4 species), and other uses (5 species). Various plant species potential as the commodity supporting ecotourism development included Abelmoschus manihot (traditional food) , Diospyros discolor (fresh local fruit), Nypa fruticans and Pandanus tectorius (local handicrafts), Phalaenopsis amabilis (ornamental plants or cutting flowers). The use of plants in Bawean Island needs to consider the sustainable use of plants and conservation approaches. Keywords: Bawean, conservation, ecotourism, local knowledge, plant diversity.

Morfologi dan Pertumbuhan Planlet Hasil Induksi Poliploidi melalui Perlakuan Kolkisin pada Kuncup Bunga Anggrek Bulan (Phalaenopsis amabilis (L.) Blume)

<br />Induction of polyploid gametes is one of useful plant polyploidization methods. Some of its benefits are to obtain triploid and tetraploid progenies at the same time by cross and self pollination. Previous research showed that some morphological characters which could be the indications of polyploidy plantlets before the analysis of chromosome number. Colchicine treatment on flower bud of diploid Phalaenopsis amabilis was conducted to determine the effect of colchicine on flower bud development, plantlets morphology and growth, and potential of polyploidy induction based on plantlets morphology. Colchicine concentrations in the experiment were 0, 50, 500, 1,000, and 2,000 mg L-1, with three days duration of treatments with aluminium foil wraps on flower buds. The results showed that high colchicine concentrations (2,000 mg L-1) inhibited flowers blooming of treated flower buds. Based on morphological characters, plantlets were classified into normal and putative polyploid plantlets. Putative polyploid plantlets from colchicine with the concentration of 50, 500, and 1,000 mg L-1 were 71.2, 86.4, and 100.0% respectively.<br /><br />Keywords: colchicine concentration, morphological characters, normal plantlets, putative polyploidy, reproductive organ<br /><br />

Polyploidy Induction of Moth Orchid (Phalaenopsis amabilis (L.) Blume) by Colchicine Treatment on Pollinated Flowers

The effectiveness of colchicine treatment at inducing polyploidy in Phalaenopsis amabilis (2n=2x=38) was investigated. Colchicine was applied on P. amabilis pollinated flowers and the development of the pods (fruit) was examined. Colchicine concentration in the experiment were 0, 50, 500, 1000, and 2000 mg L-1, with three and five days duration. The result showed that higher colchicine concentration decreased pod length and pod number. Visual screening of the seedlings obtained from colchicines treatments were classified into normal seedlings (NS), i.e. those having similar morphology to control, and putative polyploidy mutant seedlings (PMS). All colchicine-treated PMS were significantly different compared to control in the basal organ of the protocorm (BOP) length and width, leaf length, and root length and diameter. The PMS chromosome number analysis revealed that 50 mg L-1 for three or five days colchicine application and 500 mg L-1 for five days colchicine application produced 60 and 100% tetraploid mutant seedlings, respectively.

Induksi Poliploidi Phalaenopsis amabilis (L.) Blume dan Phalaenopsis amboinensis J. J. Smith dengan Kolkisin dalam Kultur In Vitro

<p>ABSTRACT</p><p>Phalaenopsis amabilis (L.) Blume and Phalaenopsis amboinensis J.J. Smith (diploid) are important in Phalaenopsis breeding. Polyploid species are needed for crossing with polyploid hybrid varities of Phalaenopsis. The objectives of this study were to obtain effective concentration of colchicine to induce polyploidy and to produce polyploid plantlets of P. amabilis and P. amboinensis. Experiment was arranged in randomized complete block design with one factor, the colchicine concentration. Protocorms of P. amabilis and P. amboinensis were immersed in half strength of Murashige-Skoog (1/2 MS) liquid media added with colchicine (0; 0.5; 5; 25; 50, and 75 mg L-1) for 10 days. The results showed that higher concentration of colchicine on both species did not have significant effect on the survival of the plantlets at 24 weeks after treatment. The average number of leaves and roots of colchicine treated planlets from both species were less than the control plantlets. Immersing protocorm in colchicine at concentration of 50 mg L-1 for 10 days was effective in inducing polyploid plantlets of P. amabilis and P. amboinensis with the frequency of 33.3% and 40%, respectively. Polyploid plantlet has larger stomata size and lower stomata density than the diploid ones.</p><p>Keywords: chromosome number, colchicine, polyploid, protocorm, stomatal density, stomatal size</p>

Selection of Phalaenopsis amabilis L. Blume Orchid Resistance to Hygromycin

Examination of Phalaenopsis amabilis orchid resistance to hygromycin antibiotic is an important step to doprior to Agrobacterium-mediated genetic transformation in this orchids using Hygromycin phosphotransferase(HPT) gene as a selection marker in the T-DNA that harboring a desired gene to be transfered. We exposedthe plant on hygromycin containing medium. The experiment was conducted using 6 weeks old P. amabilisprotocorms. These protocorms were subcultured onto NP medium supplemented with various concentrationof Hygromycin (0, 5, 10, 20, 1nd 40 mg/l). The number of survival protocorms were examined every week for4 weeks after subcultured (WAS). The resistancy of hygromycin was calculated as ratio of death protocormsper total protocorms). The result showed that 10 mg/l hygromycin with 1 weeks of application caused deathclose to LD 50. This data indicate that P. amabilis resistance to hygromycin treatment on the appropriateconcentration 10 mg/l, and this concentration can be used for other purposes in orchid system.

INVENTARISASI KEANEKARAGAMAN ANGGREK (Orchidaceae) DI HUTAN RESORT WAY KANAN BALAI AMAN NASIONAL WAY KAMBAS SEBAGAI SUMBER INFORMASI DALAM MELESTARIKAN PLASMA NUTFAH

Abstract: Orchid is one of the plants that have different characteristics in each species and its habitat. Orchids (Orchidaceae) or nation is a group that is protected by Government Regulations Number. 7 Th.1999. It is necessary to look at the type of data collections and save it, and look at the characteristics of tropical forests in the Resort Way Kanan. This research aims to inventory orchid the found in Forest Resort of Way Kanan. This research was conducted in December 2014 until January 2015, using purposive sampling method and using techniques Line transeck, cut the line following the path of the Post traking Right Way to the SRS (Sumatran Rhino Sanctuary), in this research using 4 stations between stations within 2km. The results of this study were obtained 23 species of orchids with 9 genera and 14 species include: the genus Dendrobium, Dendrobium creatceum, Dendrobium crumenatum, genus Grammatophyllum, Grammatophyllum scriptum BL, Agrostophyllum genus, the genus Cymbidium, Cymbidium chloranthum, Cymbidium hartinahianum, Calanthe genus, the genus Bulbophyllum, Arachnis sp, Angraecum didieri, Ascocentrum miniatum, Ascocentrum aureum, Coelogyne foerstermanni, golden shower Oncidium, Phalaenopsis amabilis, Spathoglottis sp. Having analyzed the data obtained it has a value equal to 46% of species density and relative density of 88.36% with a diversity index value of 1.3, based on the criteria of the Shannon-Wienner in Fachrul abundant species diversity can be expressed with the criteria being. The results of this study compiled into resources in conserving germplasm in the form of books and banner summary information, because there are some orchids are found in Forest Resort Way Kanan Balai Way Kambas National Park in Government Regulations Number. 7 Th.1999 is a protected species of orchids. Orchid is a protected plant species, therefore it is necessary for us to maintain and preserve it.

INDUCTION OF IN VITRO FLOWERING OF INDONESIAN WILD ORCHID, Phalaenopsis amabilis (L.) Blume

<p>Orchids are generally cultivated for their flower. To induce flower initiation in Phalaenopsis amabilis orchid, genetic and physiological approaches were developed. Genetic modification by insertion of P. amabilis Flowering Locus T (PaFT) gene driven by Ubiquitin promoter into orchid genome using Agrobacterium tumefaciens, whereas physiological approach was conducted by the use of growth regulators: N6­benzyladenine (BA) (1, 3, 9) mg.L-1 or gibberellic acid (GA3) (5, 10, 15) mg.L-1 alone or in combination in culture medium. Orchid seeds were sown on New Phalaenopsis (NP) medium for 8 weeks, then subcultured on NP liquid medium + BA + GA3 with shaking for 9 weeks. Developping protocorms were spread on NP solid medium, then supplemented with NP liquid medium + BA + GA3 (5:2). Cultures were maintained at 25oC with a photoperiod of 8 hrs light/16 hrs dark. For genetic transformation, 3 weeks old protocorms were immersed overnight in cultures of A. tumefaciens with T-DNA harboring Ubipro::PaFT and Hygromycin phosphotransferase (HPT) gene as selectable marker. Phenotypic analysis was carried out from 5-20 plants, each of them was observed for leaf and root number and lengths, comparing with untreated plants. Shoots with normal phenotype were generated from all treatments. RT-PCR analysis from 3 plants each of 4 weeks-24 months old-WT plants, 6 months old phytohormone treated plants and also 12 and 24 months old transgenic plants showed that POH1 juvenile gene transcript can be detected at juvenile stage of WT and PaFT mRNA was expressed in late stage after 6 months old WT plants. In all phytohormone treated plants and transgenic orchid both POH1 anf PaFT transcripts can be detected in 5, 12 and 24-months old plants, but no flower initiation was occurred. It indicates that post transcriptional inhibition might be occurred, and it needs to be explored.</p><p><br /><strong>Keywords</strong>: in vitro, flower, PaFT, growth regulators, Orchids</p>

Biogeography of the Phalaenopsis amabilis species complex inferred from nuclear and plastid DNAs.

BackgroundPhalaenopsis is one of the important commercial orchids in the world. Members of the P. amabilis species complex represent invaluable germplasm for the breeding program. However, the phylogeny of the P. amabilis species complex is still uncertain. The Phalaenopsis amabilis species complex (Orchidaceae) consists of subspecies amabilis, moluccana, and rosenstromii of P. amabilis, as well as P. aphrodite ssp. aphrodite, P. ap. ssp. formosana, and P. sanderiana. The aims of this study were to reconstruct the phylogeny and biogeographcial patterns of the species complex using Neighbor Joining (NJ), Maxinum Parsimony (MP), Bayesian Evolutionary Analysis Sampling Trees (BEAST) and Reconstruct Ancestral State in Phylogenies (RASP) analyses based on sequences of internal transcribed spacers 1 and 2 from the nuclear ribosomal DNA and the trnH-psbA spacer from the plastid DNA.ResultsA pattern of vicariance, dispersal, and vicariance + dispersal among disjunctly distributed taxa was uncovered based on RASP analysis. Although two subspecies of P. aphrodite could not be differentiated from each other in dispersal state, they were distinct from P. amabilis and P. sanderiana. Within P. amabilis, three subspecies were separated phylogenetically, in agreement with the vicariance or vicariance + dispersal scenario, with geographic subdivision along Huxley’s, Wallace’s and Lydekker’s Lines. Molecular dating revealed such subdivisions among taxa of P. amabilis complex dating back to the late Pleistocene. Population-dynamic analyses using a Bayesian skyline plot suggested that the species complex experienced an in situ range expansion and population concentration during the late Last Glacial Maximum (LGM).ConclusionsTaxa of the P. amabilis complex with disjunct distributions were differentiated due to vicariance or vicariance + dispersal, with events likely occurring in the late Pleistocene. Demographic growth associated with the climatic oscillations in the Würm glacial period followed the species splits. Nevertheless, a subsequent population slowdown occurred in the late LGM due to extinction of regional populations. The reduction of suitable habitats resulted in geographic fragmenttation of the remaining taxa.Electronic supplementary materialThe online version of this article (doi:10.1186/s12870-015-0560-z) contains supplementary material, which is available to authorized users.

Kandungan Klorofil Daun Planlet Anggrek Bulan (Phalaenopsis amabilis (L.) Bl.) Hasil Seleksi dengan Asam Salisilat secara In vitro

Phalaenopsis amabilis is one of Indonesias national flowers. P.amabilis has beautifulflowers and exciting and high economic value. However, there are some constraints in its development, one of which is Fusarium wilt caused byFusarium oxysporum, for this is not maximized in handling. One of effective procedure to control the disease is to use salicylic acid as a selective agent resistance to the disease. Study of the effect of the addition of salicylic acid(concentrations 15, 30, 45 and 60 ppm) have been studied in vitro in mediumVW(Vacint and Went) on the content ofchlorophyll a, b, and total leaf plantlets P.amabilis. The research was implemented in tissue culture laboratory, Department of Biology, Faculty of MIPA, University of Lampung from February to March 2015. This study used a completely randomized design with 3 replications. Analysis of variance and LSD test performed at 5% significance level. Calculation and extraction of chlorophyll was conducted by Harbourne method(1987). The absorbance was measured with a (Shimadzu UV 800)spectrophotometer at wavelength of 663 and 646 nm. The results showed that the content of chlorophyll a, b, and total plantlets of P. amabilis leaves significantly increased at the concentration of salicylic acid in the medium VW 15, 30, 45 and 60 ppm compared with control(0 ppm).Key words : Phalaenopsis amabilis plantlets, chlolophyll, salicylic acid, VW medium.

First Report of Dickeya solani Causing Soft Rot in Imported Bulbs of Hyacinthus orientalis in China

A bacterial pathogen, Dickeya solani, emerged as a major threat to potato (Solanum tuberosum) production in Europe in 2004 and has spread to many potato-growing regions via international trade. In December 2013, soft rot symptoms were observed in hyacinth (Hyacinthus orientalis) bulbs imported from the Netherlands into China at Ningbo Port. Diseased bulbs gave off an offensive odor. The base and internal parts of diseased bulbs rotted, and the margins of diseased tissues showed brown discoloration. Isolation on nutrient agar glucose (NAG) medium resulted in dominating colonies of characteristic "fried egg" morphology (1). One colony was chosen for further investigation and tentatively named "isolate 6165-3." Under microscopic visualization after gram stain, the cells of isolate 6165-3 were gram-negative, motile, and rod shaped. The isolate was then identified as a member of genus Dickeya using the Biolog GN microplate. The 16S rRNA, recA, and dnaX sequences of isolate 6165-3 were subsequently determined and deposited in GenBank with accession numbers KM405240, KM405241, and KM405242, sharing 99% (16S rRNA), 100% (recA), and 100% (dnaX) nucleotide identity with those of known D. solani isolates, respectively. By this means, the isolate 6165-3 was identified as D. solani (1,2). To confirm the pathogenicity of the isolate, four plants each of 30-day-old hyacinth, 14-day-old potato, and 60-day-old moth orchid (Phalaenopsis amabilis) were inoculated with suspensions of the isolate with a concentration of 108 CFU/ml in sterile water by stabbing. Plants were incubated in a climate chamber at 28°C during the day and 24°C during the night with a relative humidity of 93% and a photoperiod of 12/12 h. Plants inoculated with sterile water were included as negative controls. After 2 or 3 days, typical symptoms such as water-soaked lesions and soft rot developed around the inoculation point, while the negative controls remained symptomless. Koch's postulates were fulfilled by re-isolating bacteria from lesions, which had identical sequence and morphology characters with the inoculated isolate. This is the first report of intercepted D. solani on hyacinth bulbs imported from the Netherlands into China, indicating that D. solani can spread via hyacinth. Further spread of the pathogen into potato production might lead to immeasurable economic consequences for China.

EFEKTIFITAS RHIZOCTONIA MIKORIZA DALAM MENGINDUKSI KETAHANAN ANGGREK PHALAENOPSIS AMABILIS TERHADAP FUSARIUM SP.

This study examines the effectiveness of mycorrhizal Rhizoctonia resistance induction in Phalaenopsis amabilis against Fusarium sp. Fusarium solani is known as pathogens that attack many orchids P. amabilis (Chung et al., 2011) compared to other pathogenic fungi. Attack of Fusarium sp. will cause rot and yellow colored leaves. Until now there has been known as a biological control orchid against Fusarium sp. In this study tested the endurance locations in Sleman and Surakarta to see the effectiveness of a good orchid growth induced by Rhizoctonia mycorrhizal or not to attack by Fusarium sp. The results of the study showed that mycorrhizal Rhizoctonia able to inhibit the attack of Fusarium sp. It is shown by the value of the index of disease resistance  (DSI) in P. amabilis orchid mycorrhizal Rhizoctonia induced lower than that not induced. Mycorrhizal Rhizoctonia induction results in Sleman provide a more real than mycorrhizal Rhizoctonia induction in Surakarta.