scholarly journals Effects of culture medium and temperature on fungal growth of Mycena and marasmioid isolates and in vitro symbiotic culture in mycoheterotrophic orchid, Gastrodia pubilabiata

2021 ◽  
Author(s):  
Akane Shimazaki ◽  
Kana Higaki ◽  
Kento Rammitsu ◽  
Yuki Ogura-Tsujita
Keyword(s):  
Seed Germination ◽  
Culture Medium ◽  
Organic Materials ◽  
Agar Medium ◽  
Leaf Disc ◽  
Japanese Cedar ◽  
Malt Extract ◽  
Fungal Isolates ◽  
Mycorrhizal Association

In vitro symbiotic culture of Gastrodia pubilabiata seeds were conducted with the fungal isolates from G. pubilabiata roots. We obtained five fungal isolates, which belonged to Mycena, Marasmiaceae and Omphalotaceae. Firstly, optimal temperature and culture medium for subculture of these fungal isolates were examined. All five isolates grew the fastest on malt extract agar medium. Mycelial growth rate was highest at 25 °C between 10 °C and 40 °C. Secondly, we evaluated suitable culture vessels and organic materials for symbiotic culture. Seeds germinated well in petri dishes with Quercus leaf disc on water agar medium, and the seed germination process was well observed without dense mycelium. The most developed seedlings were found in glass bottles filled with Japanese cedar leaves, but densely grew mycelium prevent accurate seedling counts. Leaves of Quercus, Japanese cedar and bamboo were used as organic materials for symbiotic culture. All three leaves induced seed germination with Mycena and Marasmiaceae fungi, but material types affected subsequent seedling growth. Our method will contribute to understanding the mycorrhizal association of Gastrodia species and also other mycoheterotrophic plants.

scholarly journals In Vitro Seed Germination and Embryo Culture in Nothapodytes foetida (Wight) Sleumer

2015 ◽  
Vol 48 ◽  
pp. 23-31 ◽  
Author(s):  
S. Kaveri ◽  
Rao Srinath
Keyword(s):  
Seed Germination ◽  
Embryo Culture ◽  
Filter Paper ◽  
Agar Medium ◽  
Ms Medium ◽  
Mature Embryos ◽  
Nothapodytes Foetida ◽  
Half Strength ◽  
Paper Bridge

In vitro seed germination and embryo culture have been achieved in Nothapodytes foetida, this plant is known for its rich source of anticancer drug i. e., Camptothecin. In present study both normal and decoated seeds were subjected to different treatments viz., H2O, GA3, H2O2, H2SO4, chlorine water and mechanical scarification, further these were germinated on water agar medium (WA), filter paper bridge (FB), half strength MS (HMS) and full strength MS (FMS) medium. The highest percentage (69%) of germination was achieved from decoated seeds treated with 10mg/L GA3 and germinated on Filter Paper Bridge. And for embryo culture mature embryos were inoculated on MS medium containing various combination and concentrations of cytokinins (BAP, Kn and TDZ) and auxin (IAA and NAA) for rapid conversion into a plantlet. Among the different combinations of growth regulators; highest frequency (100%) of plantlet conversion was obtained on MS medium containing Kn (1.0mg/L) and NAA (0.2mg/L).

scholarly journals A Review on Asymbiotic Seed Germination in Orchids through Plant Tissue Culture

2021 ◽  
Vol 1 (1) ◽  
Author(s):  
Purnima Paramanik ◽  
◽  
Dipak Kar ◽  
Subrata Raha ◽  
◽  
...  
Keyword(s):  
Seed Germination ◽  
Mineral Nutrients ◽  
Symbiotic Association ◽  
Organic Additives ◽  
Asymbiotic Seed Germination ◽  
Orchid Species ◽  
Mycorrhizal Association ◽  
Commercially Important

Orchid seeds are very small, dust like in appearance, fusiform in shape, lacking endosperm and have undifferentiated embryo covered by transparent seed coat. Mycorrhizal association is required for seed germination of naturally growing orchids. In this symbiotic association, orchid species are dependent on mycorrhizal association for supply of mineral nutrients. In in-vitro condition, such demand of minerals may be compensated by external supply of sugar and mineral nutrients that are required for seed germination of orchid. Several orchids are responded by this asymbiotic seed culture and have commercial importance. Therefore, formulation of efficient in-vitro protocol is important for commercially important orchid species as well as endangered orchid species for conservation. This review paper is focused on various aspects of asymbiotic seed germination of orchids and the role of organic additives in successful seed germination.

scholarly journals ASYMBIOTIC GERMINATION OF CYPRIPEDIUM CALCEOLUS VAR. PARVIFLORUM (ORCHIDACEAE) IN VITRO.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 618d-618
Author(s):  
Chin-Chang Chu ◽  
Kenneth W. Mudge
Keyword(s):  
Seed Germination ◽  
Suspension Culture ◽  
Agar Medium ◽  
Germination Rate ◽  
Coconut Water ◽  
Mature Seed ◽  
Orchid Species ◽  
Cypripedium Calceolus ◽  
Chilling Treatment

Seeds from mature seed pods of Cypripedium calceolus var. parviflorum were germinated on 1/4 MSMO (Sigma) + 100ml/l coconut water + 1% sucrose +/- 8g/l agar (pH 6.0), and with or without prechilling at 5C for 8 weeks. Protocorm with apex (stage 3) was use as an index of germination. Seeds sown on agar medium withou chilling treatment resulted in a 40% germination rate in 120 days but the germination was very uneven. Seeds germinated on agar medium with prechilling developed more synchronously with 92% germination in 60 days (ie. about 120 days after sowing). Suspension culture of seeds without prechilling resulted in 85% germination after 90 days. The synchronization of seed germination in suspension culture was intermediate between that on agar with and without prechilling. Protocorms germinated in suspension culture appeared morphologically identical to those germinated on agar medium. All stage 3 protocorms developed further on the same agar medium in darkness. Both agar and suspension culture in media containing coconut water provided reliable seed germination methods for this orchid species.

scholarly journals Enhanced Seed Germination and Seedling Growth of Tillandsia Eizii In Vitro

HortScience ◽  
2003 ◽  
Vol 38 (1) ◽  
pp. 101-104 ◽  
Author(s):  
Kimberly A. Pickens ◽  
James M. Affolter ◽  
Hazel Y. Wetzstein ◽  
Jan H.D. Wolf
Keyword(s):  
Seed Germination ◽  
In Vitro Culture ◽  
Seedling Growth ◽  
Culture Medium ◽  
Habitat Destruction ◽  
Light Conditions ◽  
Physiological Constraints ◽  
Large Numbers ◽  
In The Wild

Tillandsia eizii is an epiphytic bromeliad that due to over-collection, habitat destruction, and physiological constraints has declined to near threatened status. This species exhibits high mortality in the wild, and seed are characterized by low percentages of germination. As a means to conserve this species, in vitro culture protocols were developed to enhance seed germination and seedling growth. A sterilization protocol using 70% ethanol for 2 minutes followed by 2.6% NaOCl for 40 minutes disinfested seed and promoted seedling growth. Sucrose incorporated into the culture medium had no effect on germination or growth, while NAA inhibited growth, but not germination. Cultures maintained under a 16-hour photoperiod at 22 °C exhibited greater growth than those grown at 30 °C. Seed that germinated in the dark remained etiolated and failed to develop even after transfer to light conditions. Plants grown in vitro were successfully acclimatized and transferred to the greenhouse. Over 86% survival and rapid growth were obtained with either an all-pine-bark medium, or a mixture of 2 redwood bark: 2 fir bark: 2 potting mix: 1 perlite. This demonstrated that in vitro culture of seed may be used to rapidly produce large numbers of T. eizii, and thus can be used for the conservation and reintroduction of this species.

scholarly journals Fungicide Seed Treatment Effects on Seedling Damping-off of Pumpkin Caused by Phytophthora capsici

Plant Disease ◽  
2003 ◽  
Vol 87 (1) ◽  
pp. 63-68 ◽  
Author(s):  
M. Babadoost ◽  
S. Z. Islam
Keyword(s):  
Seed Germination ◽  
Untreated Control ◽  
Mycelial Growth ◽  
Seed Treatment ◽  
Culture Medium ◽  
Phytophthora Capsici ◽  
Field Trials ◽  
Seedling Vigor ◽  
Damping Off

Apron XL LS (mefenoxam) and Allegiance FL (metalaxyl) were highly inhibitory to growth of mycelium of Phytophthora capsici in vitro. Effective dose (ED50) of mefenoxam and metalaxyl for 50% inhibition of mycelial growth, for all five isolates of P. capsici tested, was 0.98 and 0.99 μg a.i./ml of culture medium, respectively. For mefenoxam at 200 μg a.i./ml, sporangium and zoospore germination were reduced by 92 and 96%, respectively, and 21 and 24%, respectively, for metalaxyl. In greenhouse studies, seed treatment with mefenoxam (0.42 ml of Apron XL LS/kg of seed) and metalaxyl (0.98 ml of Allegiance FL/kg of seed) significantly reduced pre- and post-emergence damping-off of seedlings caused by P. capsici in three pumpkin cultivars (Dickinson, Hybrid-401, and Hybrid-698) tested. Thirty-one days after seeding, at inoculum levels of 0, 90, 600, 1,400, and 4,000 CFU/g of soil, the average seedling stands for mefenoxam treatment were 98.4, 93.8, 88.3, 77.8, and 64.8%; for metalaxyl, 99.1, 85.3, 85.8, 73.5, and 59.3; and for the untreated control, 97.5, 55.2, 45.7, 37.0, and 22.9%, respectively. In field trials, the average seedling stands 35 days after seeding were 76.7, 74.7, and 44.9% for mefenoxam, metalaxyl, and untreated control, respectively. Seed treatment with mefenoxam or metalaxyl did not have any significant effect on either seed germination or seedling vigor.

scholarly journals IN VITRO SEED GERMINATION AND PLANT GROWTH OF “CABEÇA-DE-FRADE” (CACTACEAE)

2021 ◽  
Vol 34 (1) ◽  
pp. 1-8
Author(s):  
SHEILA VITÓRIA RESENDE ◽  
ALONE LIMA-BRITO ◽  
GABRIELA TORRES- SILVA ◽  
JOSÉ RANIERE FERREIRA DE SANTANA
Keyword(s):  
Seed Germination ◽  
Plant Growth ◽  
Culture Medium ◽  
Ornamental Plants ◽  
In Vitro Cultivation ◽  
Apical Segment ◽  
Aerial Portion ◽  
Low Concentrations ◽  
Chemical Sterilization

ABSTRACT The genus Melocactus (“cabeça-de-frade”) comprises 32 species in Brazil, of which M. glaucescens and M. paucispinus are threatened with extinction. The present work evaluated the effects of different concentrations of Murashige & Skoog (MS, MS/2 and MS/4) culture medium and sucrose (15 g L-1 and 30 g L-1) on in vitro seed germination and plant growth of M. glaucescens and the efficiency of sterilization with sodium hypochlorite (NaOCl) on in vitro seed germination and plant growth of M. glaucescens when using seeds and M. glaucescens and M. paucispinus when using apical segment of cladode. In M. glaucescens, the final germination at the different MS and sucrose concentrations varied between 53.5 and 68.1% and the best results for in vitro growth were observed with the lowest mineral salt (MS/2 and MS/4) and sucrose (15 g L-1) concentrations, with lengths of the aerial portion of 9.70 and 10.76 mm, respectively. There was no difference in seed germination and plant growth in chemical and autoclave medium. It is concluded that the use of chemical sterilization with NaOCl at low concentrations of salts (MS/2 and MS/4) and sucrose (15 g L-1) are quite advantageous for producing ornamental plants germinated in vitro and/or apical segment of cladode of M. glaucescens andM. paucispinus, representing a reduction of costs for in vitro cultivation of this species.

scholarly journals Indução e caracterização morfológica e bioquímica de calos de Hyptis leucocephala (Lamiaceae)

2012 ◽  
Vol 12 (1) ◽  
pp. 151 ◽  
Author(s):  
Danilo Marcelo Santos Pereira ◽  
Lenaldo Muniz De Oliveira ◽  
Cristina Ferreira Nepomuceno ◽  
José Raniere Ferreira De Santana ◽  
Marília Lordelo Cardoso Silva ◽  
...  
Keyword(s):  
Yeast Extract ◽  
Callus Induction ◽  
Culture Medium ◽  
Callus Formation ◽  
Semiarid Region ◽  
In Vitro Cultivation ◽  
Malt Extract ◽  
Organic Additives ◽  
Brazilian Semiarid

Hyptis leucocephala is an aromatic herb endemic to the Brazilian semiarid region, with antimicrobial, antifungal,cytotoxic, anti-inflammatory and anti-HIV properties. In vitro cultivation makes the production of high genetic andsanitary quality cuttings possible for this species, allowing its sustainable exploitation. In the present study, the effect of 6-benzilaminopurine (BAP) and naftalenoacetic acid (NAA) on callus induction in leaf segments of this species anddifferent organic additives to WPM (Wood Plant Medium) for the callus induction was evaluated. The combination ofBAP and NAA allowed the induction of compact and undifferentiated calluses. The highest rate of callus formation andgrowth was obtained with the combination of 8.88 μM BAP and 21.48 μM NAA. Supplementing the culture medium withcoconut water, malt extract and yeast extract did not induce friable calluses. The levels of total sugars, reducing sugars,protein, and amino acids were reduced in calluses maintained in culture medium supplemented with coconut water, maltextract and yeast extract.

scholarly journals REGENERASIRUMPUT LAUT Kappaphycus alvarezii (Doty) MELALUI INDUKSI KALUS DAN EMBRIO DENGAN PENAMBAHAN HORMON PERANGSANG TUMBUH SECARA IN VITRO

2009 ◽  
Vol 4 (1) ◽  
pp. 39
Author(s):  
Emma Suryati ◽  
Sri Rejeki Hesti Mulyaningrum
Keyword(s):  
Acetic Acid ◽  
Growth Regulator ◽  
Culture Medium ◽  
Solid Medium ◽  
Agar Medium ◽  
Kappaphycus Alvarezii ◽  
Indol Acetic Acid ◽  
High Quality Seed ◽  
Medium Culture

Regenerasi rumput laut Kappaphycus alvarezii dilakukan dalam rangka penyediaan benih yang bermutu dan mempunyai keunggulan melalui induksi kalus dan embrio dengan penambahan hormon pertumbuhan yang diintroduksi ke dalam media kultur yang dapat memacu induksi kalus dan penebalan pigmen rumput laut. Media kultur yang digunakan adalah media Conwy padat dengan penambahan agar 0,8%-1,6%. Hormon perangsang tumbuh yang digunakan untuk memacu pertumbuhan kalus dan filamen embrio yaitu IAA (Indol acetic acid), kinetin, dan auxilin dengan konsentrasi berkisar 0,4-1 mg/L. Embrio yang dihasilkan merupakan anakan yang mempunyai sifat yang sama dengan induknya. Sintasan dan perkembangan embrio yang paling baik yaitu dengan penambahan IAA dengan konsentrasi 0,4 mg/L pada media padat. Pembentukan anakan dilakukan dengan mengiris embrio dan menumbuhkan pada media cair yang diperkaya dengan hormon yang sama. Pemeliharaan anakan pada media kultur dilakukan hingga mencapai ukuran 2-3 cm.Regeneration of seaweed Kappaphycus alvarezii was done to provide high quality seed through callus and embryo induction using plant growth regulator which was introducted to the culture medium. This growth regulator can stimulate the callus induction procces and thickening the seaweed pigment. Applied medium culture was agar medium with 0.8%-1.6% concentration enriched with Conwy and the applied growth regulators were IAA (Indol acetic acid), kinetin dan auxilin with 0.4-1 mg/L concentration range. Resulted embryo has the same characteristics with the stock. The best survival rate and embryo growth was IAA treatment with 0.4 mg/L concentration. Formation of embryo was done by transferring them from solid medium to the liquid one with the same growth regulator treatment. The nursery of the seed in culture medium was carried out until it has reached 2-3 cm in size.

scholarly journals Effect of agar concentration in culture medium on in vitro development of potato plants

2020 ◽  
Vol 9 (7) ◽  
pp. e542974571
Author(s):  
Cinthia Kutz de Matos ◽  
Camila Eduarda Loli Pereira ◽  
Leonardo Balena ◽  
Jackson Kawakami
Keyword(s):  
Culture Medium ◽  
Agar Medium ◽  
Dry Mass ◽  
Culture Technique ◽  
Potato Cultivars ◽  
Semisolid Medium ◽  
Potato Plants ◽  
Best Response ◽  
Vitro Development

In vitro propagation and tissue culture technique is an alternative for obtaining healthy potato seeds. However, data found in the literature differ on the optimal consistency of the culture medium to be used in vitro. The aim in this research was to look at the adaptation of potato cultivars to medium with different agar concentrations. The experimental design was entirely randomized in a factorial scheme. The experiment tested the effect of 3 agar medium concentrations (solid, semisolid and liquid) on 6 cultivars (Agata, Bel, Camila, Catucha, Clara and Eliza), with 5 repetitions. We quantified from the plantlets, fresh and dry mass of roots and shoot, the number of nodes, the multiplication capacity and vigor. The study showed that potato cultivars respond significantly to different concentrations of agar and their best response was in the semisolid medium.

IN VITRO PROPAGATION OF OTTAWA 3 APPLE ROOTSTOCK

1983 ◽  
Vol 63 (1) ◽  
pp. 183-188 ◽  
Author(s):  
E.-C. PUA ◽  
CALVIN CHONG ◽  
G. L. ROUSSELLE
Keyword(s):  
Culture Medium ◽  
Agar Medium ◽  
Shoot Proliferation ◽  
Normal Growth ◽  
Naphthalene Acetic Acid ◽  
Shoot Cultures ◽  
Apple Rootstock ◽  
In Vitro Proliferation ◽  
In Vitro Shoot Proliferation

Methods were developed for obtaining normal shoot cultures and for rapid in vitro proliferation and rooting of Ottawa 3 apple rootstock from meristem tips. The presence of naphthalene acetic acid (NAA) and benzyladenine (BA), both at concentrations of either 0.5 or 1.0 mg/L in the culture medium, was most effective for in vitro shoot proliferation, but growth was abnormal. Normal growth was achieved when shoots were cultured with a combination of 0.5 mg/L NAA, 0.5 mg/L BA and 5.0 mg/L gibberellic acid. One hundred percent rooting was achieved after 2 wk on agar medium supplemented with 6.25 mg/L indole butyric acid.Key words: Tissue culture, Malus, meristem cloning, growth regulators

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