Development and Evaluation of Laboratory Bioassays to Study Powdery Mildew Pathogens of Phlox In Vitro

The genus Phlox consists of approximately 65 species that include some of the most prevalent ornamental plants in the temperate zone. These popular ornamentals are extremely susceptible to powdery mildew (PM) caused by the biotrophic fungi Golovinomyces magnicellulatus and Podosphaera sp. In this study, we used Phlox paniculata and P. glaberrima to develop a set of laboratory tools to study these pathogens in vitro, including a detached leaf and a micropropagated plantlet bioassay. We assessed pathogen growth under different experimental conditions, which included the use of four different media variations (1/2 MS medium amended with benzimidazole and tetracycline), three ages of pathogen culture (14, 18, and 22 days), three phenological stages of the host tissue (1st, 3rd, and 5th node leaves), placement of inoculum on both leaf surfaces (abaxial and adaxial), and three different inoculation techniques (single spore transfer, colony tapping, colony brushing). Detached P. paniculata leaves were successfully maintained on benzimidazole-amended 1/2 MS medium for up to 3 weeks. For both pathogens, the use of 18-day-old cultures resulted in a higher number of larger, higher sporulating colonies compared with 1-4 and 22-day-old cultures. The adaxial side of 3rd node leaves supported statistically significant more fungal growth compared with the adaxial side of 1st and 5th node leaves. Both pathogens also successfully infected micropropagated plantlets of P. glaberrima. These newly developed tools should facilitate in vitro studies on PM of Phlox and possibly be applicable to other ornamental species attacked by the same fungi.

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The effect of BA and GA3 on the shoot multiplication of in vitro cultures of Polish wild roses

Folia Horticulturae ◽

10.2478/v10245-011-0022-5 ◽

2011 ◽

Vol 23 (2) ◽

pp. 145-149 ◽

Cited By ~ 5

Author(s):

Bożena Pawłowska

Keyword(s):

Relative Humidity ◽

Ornamental Plants ◽

Shoot Multiplication ◽

In Vitro Cultures ◽

Multiplication Rate ◽

Ms Medium ◽

Naturally Occurring ◽

Dog Rose ◽

The University

The effect of BA and GA3on the shoot multiplication ofin vitrocultures of Polish wild rosesThe experiment was conducted using five species of roses naturally occurring in Poland:Rosa agrestis(fieldbriar rose),R. canina(dog rose),R. dumalis(glaucous dog rose),R. rubiginosa(sweetbriar rose), andR. tomentosa(whitewooly rose), from thein vitrocollection of the Department of Ornamental Plants of the University of Agriculture in Kraków. We examined the effect of cytokinin BA (1-10 μM) added to an MS medium (Murashige and Skoog 1962) on auxiliary shoot multiplication. The second group of test media contained BA (1-5 μM) and gibberellin GA3(0.3-1.5 μM). The cultures were maintained at a phytotron temperature of 23/25°C (night/day), 80% relative humidity, with a 16-hour photoperiod and PPFD of 30 μmol m-2s-1, and cultured in five-week cycles. The highest multiplication rate was obtained forR. caninaandR. rubiginosa(4.1 shoots per one explant) andR. dumalis(2.9 shoots per one explant), when shoots were multiplied on an MS medium supplemented with 1 μM BA and 1.5 μM GA3. Multiplication was the weakest inRosa tomentosaindependent of the medium used.

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Picloram-induced enhanced callus-mediated regeneration, acclimatization, and genetic clonality assessment of gerbera

Journal of Genetic Engineering and Biotechnology ◽

10.1186/s43141-021-00269-1 ◽

2021 ◽

Vol 19 (1) ◽

Author(s):

Saikat Gantait ◽

Manisha Mahanta

Keyword(s):

Acetic Acid ◽

Ornamental Plants ◽

Dry Weight ◽

Delayed Response ◽

Ms Medium ◽

Semisolid Medium ◽

Inter Simple Sequence Repeats ◽

Dichlorophenoxy Acetic Acid ◽

Tea Leaf

Abstract Background Gerbera jamesonii Bolus ex Hooker f. (African daisy) is listed among the top five most important ornamental plants in the global floricultural industry. To satisfy its demand, the floriculture industry relies on reproducible and effective propagation protocol while retaining the genetic uniformity of G. jamesonii. The present study, for the first time, reports the potential of picloram for enhanced induction of organogenic calli from leaves of G. jamesonii and its high-frequency indirect regeneration. Results The fastest induction of calli with maximum fresh and dry weight was recorded in the Murashige and Skoog (MS) semisolid medium supplemented with 1 mg/l picloram. In addition, callus induction was observed in 2,4-dichlorophenoxy acetic acid- and α-napthaleneaceticacid-supplemented media but with delayed response and reduced fresh and dry weight. The proliferated calli were transferred to shoot induction media containing MS salt and 0.5–1 mg/l N6-benzylaminopurine, kinetin, or thidiazuron. A mean number of ~6 shoots per callus were developed after 5 days of culture in the MS medium supplemented with 1 mg/l kinetin, with a mean length of 5.2 cm. Successful rooting of shoots was achieved in the MS medium fortified with 1.5 mg/l indole-3-acetic acid, wherein the earliest root initiation (~5 days), as well as the maximum number (~9) and length (~4.8 cm) of roots, were recorded. Complete plantlets were primarily acclimatized in sand before being transferred to a mixed substrate (of soil, sand, tea leaf waste, and cow urine) that secured >90% survival and further growth of the plantlets. Eventually, clonal fidelity of the in vitro regenerants assessed via inter-simple sequence repeats (ISSR) primers exhibited a monomorphic banding patterns that suggested genetic integrity within the plantlets as well as with their mother plant. Conclusions The results of the present study should be of interest for commercial propagation and mutagenesis- as well as genetic transformation-related research.

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In vitro growth of Brassocattleya orchid hybrid in different concentrations of KNO3, NH4NO3 and benzylaminopurine

Horticultura Brasileira ◽

10.1590/s0102-05362011000300017 ◽

2011 ◽

Vol 29 (3) ◽

pp. 359-363 ◽

Cited By ~ 5

Author(s):

Jean C Cardoso ◽

Elizabeth O Ono

Keyword(s):

Tissue Culture ◽

Plant Tissue Culture ◽

Plant Tissue ◽

Ornamental Plants ◽

Dry Weight ◽

Ms Medium ◽

In Vitro Growth ◽

Mass Propagation ◽

Mineral Salts

One of the most important applications of plant tissue culture is mass propagation of ornamental plants. This experiment evaluated the effect of different concentrations of NH4NO3 and KNO3 and BAP on the in vitro growth of orchid hybrid Brassocattleya 'Pastoral'. Seedlings of this orchid hybrid were used as explants and cultivated in medium with mineral salts and vitamins from the MS medium (Murashige & Skoog, 1962), with the macronutrients P, Ca and Mg reduced by half, and with an addition of 25 g L-1 of sucrose, 0.1 g L-1 of myo-inositol and 1.5 g L-1 of activated charcoal. Agar-agar was added (6.5 g L-1) and the pH was adjusted to 5.8. As treatments, four concentrations of the NH4NO3 and KNO3 (2x; 1x; ½ and ¼ MS medium) and three concentrations of BAP (0.0; 0.5 and 1.0 mg L-1) were assayed. The multiplication, growth in height, fresh and dry weight and sugar level in dry weight of sprouts were evaluated. There occurred a higher growth in height with 0.25x NH4NO3 and KNO3 salts concentrations of MS medium and higher rate of multiplication with combination of NH4NO3 and KNO3 reduced by half of the MS medium concentration and 1.0 mg L-1 BAP.

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In Vitro Rooting of Capparis spinosa L. as Affected by Genotype and by the Proliferation Method Adopted During the Multiplication Phase

Plants ◽

10.3390/plants9030398 ◽

2020 ◽

Vol 9 (3) ◽

pp. 398

Author(s):

Valeria Gianguzzi ◽

Ettore Barone ◽

Francesco Sottile

Keyword(s):

Liquid Culture ◽

Solid Medium ◽

Base Material ◽

In Vitro Rooting ◽

The Other ◽

Ms Medium ◽

Fluorescent Lamp ◽

Experimental Conditions ◽

Capparis Spinosa

The in vitro rooting of three caper (Capparis spinosa L.) selected biotypes, grown in a commercial orchard on the Sicilian island of Salina (38°33′49” N), was performed using—as base material for rooting experiments—shoot explants proceeding from two different in vitro culture systems: solid medium and liquid culture in a PlantForm bioreactor (TIS). The regenerated shoots of each accession were submitted to different auxin treatments (NAA, IBA, IAA - 1 or 2 mg L−1; NAA+IBA 0.75 and 0.25 mg L−1, respectively), supplemented with sucrose or fructose (mg L−1). The highest rooting rate in terms of root percentage (67%) was reached with the explants of the selected accession ‘Sal 39’ proceeding from liquid culture in PlantForm and induced in the MS medium with sucrose, as a carbon source, supplemented with NAA 0.75 mg L−1 + IBA 0.25 mg L−1, after six days in a climatic growth chamber at 25 ± 1 °C in the dark and then placed under a cool white fluorescent lamp, with a PPFD of 35 μmol m−1 s−1 and a photoperiod of 16 h. On the other hand, poor rooting rate was generally achieved under all the tested experimental conditions with the other biotypes, ‘Sal 37’ and ‘Sal 35’, demonstrating the strong role exerted by the previously adopted proliferation method and by the genotype for successful caper in vitro rooting.

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Micropropagation of some Myrtaceae species which show potential as 'new' ornamental plants

Australian Journal of Experimental Agriculture ◽

10.1071/ea9930385 ◽

1993 ◽

Vol 33 (3) ◽

pp. 385 ◽

Cited By ~ 2

Author(s):

SS Speer

Keyword(s):

Ornamental Plants ◽

Shoot Proliferation ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Soil Culture ◽

Axillary Shoots ◽

Half Strength ◽

Roots In Vitro ◽

Chamelaucium Uncinatum

Eleven species of Australian Myrtaceae were evaluated for their ability to be cultured in vitro. Ten species produced axillary shoots (microcuttings) suitable for inducing roots in vitro. Microcuttings of 9 species successfully developed roots and were transferred to soil culture in a glasshouse, where plants grew normally. Nodal explants were grown on a modified Murashige and Skoog (MS) medium supplemented with varying concentrations of 6-benzylaminopurine (BAP), to study shoot proliferation. Beaufortia heterophylla Turcz. explants did not respond to BAP, and all explants eventually died. The rate of shoot proliferation for the other species varied according to BAP concentration. Microcuttings of 10 species were grown on a modified half-strength MS medium supplemented with varying concentrations of the auxins indole-3-butyric acid (IBA) and naphthaleneacetic acid (NAA), to induce root formation. An increase in root number and an associated decrease in root length was observed as the concentration of IBA and NAA was increased. Verticordia muelleriana E. Pritzel did not develop roots in any treatment. Chamelaucium uncinatum Schauer cv. Purple Pride, Kunzea parvifolia Schauer, K. pulchella (Lindl.) A. S. George, Leptospermum rotundifolium (Maiden & Betche) F. Rodway ex Cheel, Verticordia drunzmondii Schauer, V. graizdis J. L. Drumm., V. hughanii F. Muell., V. nzonaclelpha Turcz., and V. roeii Endl. microcuttings developed roots both with and without added auxins. Roots that formed on microcuttings at higher auxin concentrations were generally thicker and slower in growth.

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PERTUMBUHAN PLANLET KRISAN (Dendranthema grandiflora Tzvelev) KULTIVAR PINK FIJI SETELAH PENAMBAHAN EKSTRAK TAUGE (Vigna radiata L.) PADA MEDIUM MURASHIGE DANSKOOG (MS) SECARA IN VITRO.

Jurnal Ilmiah Biologi dan Keanekaragaman Hayati ◽

10.23960/jbekh.v5i2.50 ◽

2019 ◽

Vol 5 (2) ◽

pp. 36-41

Author(s):

Nalindri Impitasari ◽

Endang Nurcahyani ◽

Tundjung Tripeni Handayani ◽

Yulianty Yulianty

Keyword(s):

Vigna Radiata ◽

Economic Value ◽

Ornamental Plants ◽

Natural Sciences ◽

Ms Medium ◽

Dendranthema Grandiflora ◽

Community Interest ◽

Randomized Design ◽

Completely Randomized Design

Chrysanthemum (Dendranthema grandiflora Tzvelev) is one of the important ornamental plants in Indonesia and has high economic value . This plant is known as a producer of flowers with attractive shapes and colors . Seeing the magnitude of community interest and the potential utilization of chrysanthemum , causing this plant more and more developed and cultivated . This study aims to determine the concentration of optimum mungbean sprouts extract on the growth of chrysanthemum explants in vita . The addition of mungbean sprouts extracts (Vigna radiata L .) from concentration of 0% v/v , 2% v/v , 4% v/v , 6% v/v and 8% v/v on Murashige and Skoog(MS ) medium to growth eksplan Chrysanthemum (Dendranthema grandiflora Tzvelev ) Pink Fiji cultivars have been carried out in the tissue culture laboratory of Faculty of Mathematics and Natural Sciences , University of Lampung from November to December 2017 . This study used Completely Randomized Design (RAL ) 1 factor with 5 replications . Analysis of BNT variety and test is done at 5% level . The results showed that the extract from mungbean sprouts (Vigna radiata L . ) had no effect on plantlet height , number of shoot and number of chrysanthemum (Dendranthema grandiflora Tzvelev) plantlet leaves. The addition of mungbean spourts extracts on Murashige and skoog (MS) medium show 100% live plantlet.

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Response of four medical plants of Euphorbia species in callus initiation in vitro

Journal of Biotechnology Research Center ◽

10.24126/jobrc.2014.8.1.310 ◽

2014 ◽

Vol 8 (1) ◽

pp. 66-71

Author(s):

Abedaljasim M. Aljibouri ◽

Silva A. Yakoub Zokian ◽

Ali H. Almusawi

Keyword(s):

Incubation Temperature ◽

Dry Weight ◽

Callus Cultures ◽

The Other ◽

Ms Medium ◽

Light Conditions ◽

Experimental Conditions ◽

Callus Initiation ◽

Dichlorophenoxy Acetic Acid

Callus cultures were initiated for four Euphorbia species. Nodule explants cultured on Murashige and Skoog (MS) medium, supplemented with different concentrations 0,0.5,1,1.5,2 mg/l of the auxin 2,4-Dichlorophenoxy acetic acid 2,4-D. Half of the cultures were incubated for16 hrs/day photoperiod, while the other half was incubated under complete darkness. The incubation temperature was 25±1ºC. Observations on number of nodule explants initiated callus were recorded at 2,4,6,8 weeks of culture. For callus maintenance, 50mg of callus produced were re-cultured on MS medium supplemented with different concentrations of 2,4-D 0,0.4,0.8,1.2,1.6 mg/l. Callus fresh and dry weights were recorded after 4 weeks. Results showed that nodule explants of Epeplus and Ehirta incubated under light conditions achieved the highest response to callus initiation 75-100% compared with the other species under experimental conditions. E.Helioscopia incubated under light conditions achieved the lowest response for callus initiation 25-75%. Results also showed significant differences between Euphorbia species in fresh and dry callus weights, E. Ehirta produced the highest fresh and dry weight of callus reaching 1.410 and 0.046 mg respectively. The amount of fresh and dry weight of callus produced under dark conditions was significantly higher than that produced under light conditions.

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In Vitro Regeneration of Muscari racemosum Mill. Using Twin Bulb Scales, Primary Bulbs, and Leaf Bases

ISPEC Journal of Agricultural Sciences ◽

10.46291/ispecjasvol5iss3pp714-727 ◽

2021 ◽

Vol 5 (3) ◽

pp. 714-727

Author(s):

Çiğdem Alev ÖZEL ◽

Fatma ÜNAL

Keyword(s):

Clonal Propagation ◽

In Vitro Regeneration ◽

Ornamental Plants ◽

Fold Increase ◽

Naphthaleneacetic Acid ◽

Special Importance ◽

Ms Medium ◽

Related Factors ◽

Important Center

Turkey is an important center of diversity for many plants species including bulbs, rhizomes, tubers, and other plants of high agricultural and horticultural importance. These species have a special importance as ornamental plants. However, due to urbanization and related factors, many of them are under threat. One of these species is the endemic Muscari racemosum Mill. The current study aimed to develop an efficient in vitro commercial bulblet propagation procedure using different explants. Twin-scale bulb explants were regenerated on MS medium having several doses of Kinetin+NAA (1-Naphthaleneacetic acid). The best regeneration was exhibited on 4.65 μM Kinetin+5.37 μM NAA at the end of 10 weeks with induction of 4.08 bulblets/explant with a mean diameter of 0.31 cm. The primary bulblets were cultured on MS medium having 18.60 μM Kinetin+5.37 μM NAA. About a 2.5-fold increase in the diameter of the bulbs (0.76 cm) was exhibited on the regenerated bulblets. The bulblets were regenerated on leaf bases taken from MS medium having several doses of BAP (6-Benzylaminopurine) + NAA. The regenerated bulbs were rooted on MS medium having 4.90 μM IBA (Indole-3-butyric acid) followed by their transference to a greenhouse for acclimatization. This study provided important information on commercial clonal propagation of M. racemosum and the importance of explants and growth regulators in plant regeneration.

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Micropropagation of Homalomena pineodora Sulaiman & Boyce (Araceae): a new species from Malaysia

Horticultura Brasileira ◽

10.1590/s0102-05362012000100007 ◽

2012 ◽

Vol 30 (1) ◽

pp. 39-43 ◽

Cited By ~ 2

Author(s):

Christine Stanly ◽

Arvind Bhatt ◽

Baharuddin Sulaiman ◽

Chan Lai Keng

Keyword(s):

Large Scale ◽

Ornamental Plants ◽

Basal Medium ◽

Fluorescent Light ◽

Scale Production ◽

Shoot Cultures ◽

Ms Medium ◽

Plantlet Production ◽

Mother Plants

Homalomena pineodora (family Araceae) is a species found to have impressive foliage characteristics which remain evergreen throughout the year. Therefore, H. pineodora can be grown as an ornamental plant. Generally H. pineodora needs 3-5 years to propagate and multiply. However, the demand for new ornamental plants is increasing worldwide and the quality of planting material is a basic need for boosting productivity. Therefore an efficient micropropagation protocol for large-scale production of H. pineodora was developed. In vitro shoot cultures were initiated from the rhizomatous buds on MS basal medium. The best conditions for propagating H. pineodora was found to be MS medium supplemented with 3% sucrose and 0.5 mg L-1 BA (6-benzyladenine) under 24 h of cool fluorescent light which produced an average of 3.8 shoot per explant. Presence of an auxin was not necessary for plantlet production. Liquid MS medium supplemented with 0.5 mg L-1 BA, enhanced the shoot production of H. pineodora as compared to agar-gelled medium with same composition. All the in vitro plantlets of H. pineodora were successfully acclimatized with 100% survival rate. Scanning electron microscopy confirmed the similarity of leaf microstructures between the in vitro and mother plants of H. pineodora.

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In vitro conservation of two plant species (Prunus cerasifera Ehrh. and Rubus fruticosus L.) shoot tips by encapsulation dehydration

Genetika ◽

10.2298/gensr1301001r ◽

2013 ◽

Vol 45 (1) ◽

pp. 1-10 ◽

Cited By ~ 1

Author(s):

Djurdjina Ruzic ◽

Tatjana Vujovic ◽

Radosav Cerovic

Keyword(s):

Osmotic Dehydration ◽

Alginate Beads ◽

Shoot Tips ◽

Ms Medium ◽

Experimental Conditions ◽

Prunus Cerasifera ◽

Rubus Fruticosus ◽

Acid Sodium Salt ◽

Lower Capacity

In vitro grown shoot tips of cherry plum (Prunus cerasifera Ehrh.) and blackberry ?Cacanska Bestrna? (Rubus fruticosus L.) were tested for regrowth after cryopreservation using encapsulation dehydration method. Apical, 2-3 mm long shoot tips, were encapsulated in alginate beads composed of 3, 5 and 10% (w/v) alginic acid sodium salt in calcium-free liquid Murashige and Skoog (MS) medium containing 1.0 mg l-1 benzyladenine, 0.1 mg l-1 indole-3-butyric acid and 0.1 mg l-1 gibberellic acid. Polymerization was done in liquid MS medium with 100 mM CaCl2 for 30 min at room temperature. Encapsulated shoot tips were pre-treated in liquid MS medium with 0.75 or 1 M sucrose for 24 h in growth room and dehydrated for 4 and 8 h (29% and 20% moisture content respectively) before rapid immersion in liquid nitrogen. Upon thawing which involved placing the cryovials in the air current of the laminar airflow cabinet for 2 min, the beads were directly transferred to regrowth medium. In cherry plum, osmotic dehydration in 0.75 M sucrose followed by 8-hour desiccation gave the highest regrowth (60%) of explants encapsulated in 3% and 5% alginate beads. However, in comparison with cherry plum, blackberry displayed significantly lower capacity for regrowth after cryopreservation under described experimental conditions. In this genotype, osmotic dehydration in 1 M sucrose followed by 8-hour desiccation resulted in the highest regrowth (16.7%) of explants encapsulated in 5% alginate beads. Cryopreserved shoot tips of both genotypes multiplied in the three subcultures had normal morphology and similar multiplication capacity in comparison with non-cryopreserved shoots.

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