scholarly journals Variables Affecting Shoot Growth and Plantlet Recovery in Tissue Cultures of Drug-Type Cannabis sativa L.

2021 ◽  
Vol 12 ◽  
Author(s):  
Janesse E. Holmes ◽  
Samantha Lung ◽  
Danielle Collyer ◽  
Zamir K. Punja
Keyword(s):  
Silver Nitrate ◽  
Activated Charcoal ◽  
Shoot Growth ◽  
Cannabis Sativa ◽  
Sodium Metasilicate ◽  
Nodal Explants ◽  
Meristem Culture ◽  
Ms Medium ◽  
Tissue Cultures ◽  
Drug Type

Tissue culture approaches are widely used in crop plants for the purposes of micropropagation, regeneration of plants through organogenesis, obtaining pathogen-free plantlets from meristem culture, and developing genetically modified plants. In this research, we evaluated variables that can influence the success of shoot growth and plantlet production in tissue cultures of drug-type Cannabis sativa L. (marijuana). Various sterilization methods were tested to ensure shoot development from nodal explants by limiting the frequency of contaminating endophytes, which otherwise caused the death of explants. Seven commercially grown tetrahydrocannabinol (THC)-containing cannabis genotypes (strains) showed significant differences in response to shoot growth from meristems and nodal explants on Murashige and Skoog (MS) medium containing thidiazuron (1 μM) and naphthaleneacetic acid (0.5 μM) plus 1% activated charcoal. The effect of Driver and Kuniyuki Walnut (DKW) or MS basal salts in media on shoot length and leaf numbers from nodal explants was compared and showed genotype dependency with regard to the growth response. To obtain rooted plantlets, shoots from meristems and nodal explants of genotype Moby Dick were evaluated for rooting, following the addition of sodium metasilicate, silver nitrate, indole-3-butyric acid (IBA), kinetin, or 2,4-D. Sodium metasilicate improved the visual appearance of the foliage and improved the rate of rooting. Silver nitrate also promoted rooting. Following acclimatization, plantlet survival in hydroponic culture, peat plugs, and rockwool substrate was 57, 76, and 83%, respectively. The development of plantlets from meristems is described for the first time in C. sativa and has potential for obtaining pathogen-free plants. The callogenesis response of leaf explants of 11 genotypes on MS medium without activated charcoal was 35% to 100%, depending on the genotype; organogenesis was not observed. The success in recovery of plantlets from meristems and nodal explants is influenced by cannabis genotype, degree of endophytic contamination of the explants, and frequency of rooting. The procedures described here have potential applications for research and commercial utility to obtain plantlets in stage 1 tissue cultures of C. sativa.

scholarly journals The Past, Present and Future of Cannabis sativa Tissue Culture

Plants ◽  
2021 ◽  
Vol 10 (1) ◽  
pp. 185
Author(s):  
Adrian S. Monthony ◽  
Serena R. Page ◽  
Mohsen Hesami ◽  
Andrew Maxwell P. Jones
Keyword(s):  
Tissue Culture ◽  
Disease Virus ◽  
Cannabis Sativa ◽  
Clonal Plants ◽  
Future Directions ◽  
The Past ◽  
Recent Developments ◽  
History Of ◽  
Drug Type

The recent legalization of Cannabis sativa L. in many regions has revealed a need for effective propagation and biotechnologies for the species. Micropropagation affords researchers and producers methods to rapidly propagate insect-/disease-/virus-free clonal plants and store germplasm and forms the basis for other biotechnologies. Despite this need, research in the area is limited due to the long history of prohibitions and restrictions. Existing literature has multiple limitations: many publications use hemp as a proxy for drug-type Cannabis when it is well established that there is significant genotype specificity; studies using drug-type cultivars are predominantly optimized using a single cultivar; most protocols have not been replicated by independent groups, and some attempts demonstrate a lack of reproducibility across genotypes. Due to culture decline and other problems, the multiplication phase of micropropagation (Stage 2) has not been fully developed in many reports. This review will provide a brief background on the history and botany of Cannabis as well as a comprehensive and critical summary of Cannabis tissue culture. Special attention will be paid to current challenges faced by researchers, the limitations of existing Cannabis micropropagation studies, and recent developments and future directions of Cannabis tissue culture technologies.

scholarly journals In vitro Flowering of Shoots Regenerated from Cultured Nodal Explants

2011 ◽  
Vol 39 (1) ◽  
pp. 84 ◽  
Author(s):  
Kantamaht KANCHANAPOOM ◽  
Suttinee JINGJIT ◽  
Kamnoon KANCHANAPOOM
Keyword(s):  
Multiple Shoots ◽  
Shoot Formation ◽  
In Vitro Flowering ◽  
Nodal Explants ◽  
Callus Growth ◽  
Ms Medium ◽  
Old Field ◽  
Gypsophila Paniculata ◽  
Initial Culture

A protocol for the regeneration of Gypsophila paniculata L. using nodal explants from 2-month-old field grown plants was established. The induction of multiple shoots was best obtained on Murashige and Skoog (MS) medium supplemented with 13.3 μM BA. Callus growth was observed on MS medium containing 44.3 μM BA. Calluses were transferred to MS medium supplemented with 2, 4-D (4.5, 13.5, 22.6 μM), NAA (5.3, 16.1, 26.8 μM) or BA (4.4, 13.3, 22.1 μM) for 2 months to induce shoot formation. After 6 weeks of initial culture, multiple shoots were regenerated from calluses cultured on MS medium supplemented with 13.3 μM BA. All regenerated shoots produced roots on 16.1 μM NAA containing MS medium within 4 weeks. Rooted plantlets were hardened and established in pots at 100% survival. For induction of in vitro flowering, regenerated shoots could be induced to flower efficiently when cultured on MS medium containing 13.3 μM BA and 50 g/l sucrose.

scholarly journals STANDARDIZATION OF MEDIUM FOR SYNTHETIC SEED GERMINATION OF SALVIA SCLAREA L.

2015 ◽  
Vol 2 (1) ◽  
pp. 118-120
Author(s):  
Durgha H ◽  
Ramya G ◽  
Gogul Ramanth M ◽  
Thirugnanasampandan R
Keyword(s):  
Seed Germination ◽  
Sodium Alginate ◽  
Calcium Chloride ◽  
Multiple Shoot ◽  
Nodal Explants ◽  
Synthetic Seed ◽  
Ms Medium ◽  
Shoot Induction ◽  
Multiple Shoot Induction ◽  
Salvia Sclarea

Young nodal explants (0.5-1cm) of Salvia sclarea L. was used for synthetic seed preparation.Synthetic seeds were prepared using 5% sodium alginate and 1.11% calcium chloride. Seed germination was observed on MS medium fortified with 1.4µM GA3+4.4µM BA after twenty days of culture. Further multiple shoot induction was observed after fifteen days of shootinduction.

scholarly journals Diazotrophic bacteria in the growth of micropropagated ornamental pineapple

2019 ◽  
Vol 13 (2) ◽  
pp. 269-278
Author(s):  
Adriano Bortolotti Silva ◽  
Ligiane Aparecida Florentino ◽  
Dalvana De Sousa Pereira ◽  
Paulo Roberto Correa Landgraf ◽  
Ana Carolina Rodrigues Alves ◽  
...  
Keyword(s):  
Shoot Growth ◽  
Ex Vivo ◽  
Control Treatment ◽  
In Vitro Cultivation ◽  
Ms Medium ◽  
Ex Vitro ◽  
Diazotrophic Bacteria ◽  
Bacterial Strains ◽  
Indole 3 Acetic Acid

Ornamental pineapple is a hardy plant with significant landscaping value. Tissue culture of plants is viable for producing plants with a high phytosanitary quality. However, one of the difficulties with this cultivar is the acclimatization process, which is slow and can cause losses. The objective of the present study was to verify the potential of inoculation with diazotrophic bacteria for in vitro and ex vivo growth of ornamental pineapple. A group of diazotrophic bacterial strains selected at the Universidade José do Rosário Vellano (UNIFENAS) was prioritized in this study, and the treatments included bacterial strains UNIFENAS (100-13, 100-60, 100-68, 100-153, 100-167 and 100-198). These strains were evaluated in terms of their capacity to produce indole 3-acetic acid. Subsequently, plants were cultivated in a medium composed of MS medium salts (1/4), adding 1 mL of the bacterial strain. In the control treatment, the plants were maintained in 2 mL of MS medium. 7 days after inoculation, the plants were transplanted into the MS, where they were maintained for 30 days. After in vitro cultivation, the plants were transferred to pots containing commercial Plantmax® substrate and maintained under these conditions for 60 days. The diazotrophic bacteria were able to synthesize auxins, and their inoculation promoted greater growth in vitro and ex vitro in the plants. In the acclimatization phase, the plants inoculated with UNIFENAS strains (100-60, 100-68 and 100-153) promoted a higher shoot growth, chlorophyll content and nitrate reductase enzyme activity.

scholarly journals Comparative study on meristem culture of three potato cultivars diamant, cardinal and granula and their shoot formation

2016 ◽  
Vol 1 (3) ◽  
pp. 537-544
Author(s):  
Marufa Khatun ◽  
Md Shahadat Hossain ◽  
Md Khalekuzzan ◽  
Amina Rownaq
Keyword(s):  
Comparative Study ◽  
Growth Response ◽  
Potato Cultivar ◽  
Shoot Formation ◽  
Shoot Tips ◽  
Potato Cultivars ◽  
Meristem Culture ◽  
Ms Medium ◽  
Surface Sterilization ◽  
Apical Meristems

The main aim of this study was to analyze the comparative study on meristem culture of three important potato cultivars, diamant, cardinal and granula and their shoot formation using meristem tips. Apical meristems were isolated from shoot tips of 25-35 days old field grown plants. After surface sterilization the meristems (0.3 mm) were isolated from the shoot tips. After isolation the apical meristems were placed quickly on “M” shaped filter paper bridge in culture tubes containing liquid MS medium supplemented with various concentrations and combinations of different plant growth regulators. Meristems showed their first growth response by increase in size and became greenish white in colour. They continued their growth and developed shoots with roots. In the present investigation, among the three cultivars of potato, cardinal were the best responsive cultivars for the resuming new growth of cultured meristems on MS medium supplemented with KIN (0.4 mg l-1) + GA3 (0.5 mg l-1). Cultivar diamant showed comparatively better growth response in the primary culture and granula showed better results in shoot length formation in MS0 medium. Cultivar cardinal proved to be best potato cultivar in case of meristem culture than diamant and granula cultivars.Asian J. Med. Biol. Res. December 2015, 1(3): 537-544

scholarly journals Mass Propagation of Feronia limonia L. through Tissue Culture

1970 ◽  
Vol 45 (1) ◽  
pp. 75-78 ◽  
Author(s):  
Shahina Islam ◽  
Mosfequa Zahan ◽  
Shahina Akter ◽  
Tanjina Akhtar Banu ◽  
Ahashan Habib ◽  
...  
Keyword(s):  
Tissue Culture ◽  
Plant Growth ◽  
Key Words ◽  
Multiple Shoot ◽  
Shoot Tips ◽  
Nodal Explants ◽  
Ms Medium ◽  
Ex Vitro ◽  
Mass Propagation

An efficient mass propagation method for Feronia limonia was developed from excised shoot tips and nodal explants of in vitro grown seedlings. Explants were cultured on MS medium with different conc. of NAA, Kn, IAA and BAP singly or in combinations. Highest number of micro shoots and better plant growth were obtained from these two explants on MS medium supplemented with 0.2 mg/l BAP alone. The regenerated shoots were successfully rooted on MS medium supplemented with 0.5 mg/l NAA. The in vitro raised plantlets were successfully established in soil following the formation of roots with 100% survivability under ex vitro condition. Key words: Feronia limonia; Mass propagation; Node; Shoot tips; Multiple shoot DOI: 10.3329/bjsir.v45i1.5186 Bangladesh J. Sci. Ind. Res. 45(1), 75-78, 2010

scholarly journals Micropropagation of Marsdenia brunoniana Wight & Arn. - A rare antidiabetic plant

2010 ◽  
Vol 20 (1) ◽  
pp. 7-12 ◽  
Author(s):  
A. Ugraiah ◽  
S. Karuppusamy ◽  
T. Pullaiah
Keyword(s):  
Survival Rate ◽  
Key Words ◽  
Growth Regulators ◽  
Plant Tissue ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Explants ◽  
Ms Medium ◽  
Plant Micropropagation ◽  
Number Of Shoots

Shoot multiplication of M. brunoniana Wight & Arn. was achieved from the nodal explants of mature plants using MS with different concentrations and combina-tions of growth regulators. Maximum explant response and highest number of shoots per explant was obtained on MS medium fortified with 1.0 mg/l BAP. The highest degree of shoot proliferation was found to be 90%. The combination of BAP and Kn was also found to be effective for regeneration. The regenerated shoots were successfully rooted on MS supplemented with 0.5 mg/l NAA, after sequential hardening; survival rate was 90%.  Key words: Marsdenia brunoniana, Medinal plant, Micropropagation, Conservation D. O.I. 10.3329/ptcb.v20i1.5958 Plant Tissue Cult. & Biotech. 20(1): 7-12, 2010 (June)

scholarly journals In vitro study of Tinospora cordifolia (Willd.) Miers (Menispermaceae)

2010 ◽  
Vol 6 ◽  
pp. 103-105 ◽  
Author(s):  
Aditi Singh ◽  
Saroj K Sah ◽  
Aunji Pradhan ◽  
Sabari Rajbahak ◽  
Niran Maharajan
Keyword(s):  
Medicinal Plant ◽  
Callus Induction ◽  
Shoot Growth ◽  
Tinospora Cordifolia ◽  
Nodal Segments ◽  
Naphthaleneacetic Acid ◽  
Nodal Segment ◽  
Root Induction ◽  
Ms Medium

In vitro study was carried out in an important medicinal plant Tinospora cordifolia (Willd.) Miers belonging to the family: Menispermaceae. Vegetative parts such as stem, leaf and nodal explants were excised from an elite in vivo grown mature plant and thereafter cultured on Murashige-Skoog (MS) medium supplemented with different hormonal concentrations for callus induction and organogenesis. Callus formation occurred from nodal segments, leaf and inter-node explants when planted on different combinations of hormones. Tinospora cordifolia showed response for in vitro shoot growth from the nodal segment. The best shoot growth was observed on MS medium supplemented with kinetin (1.5 mg/l). Similarly, the best result for root induction was obtained on MS medium supplemented with 6-benzylaminopurine (1.0 mg/l) and naphthaleneacetic acid (2.5 mg/l). Key-words: callus induction; explants; medicinal plant; MS medium; tissue culture.DOI: 10.3126/botor.v6i0.2918 Botanica Orientalis - Journal of Plant Science (2009) 6: 103-105

scholarly journals Highly Predictive Genetic Markers Distinguish Drug-Type from Fiber-Type Cannabis sativa L

Plants ◽  
2019 ◽  
Vol 8 (11) ◽  
pp. 496 ◽  
Author(s):  
Fidelia Cascini ◽  
Alessio Farcomeni ◽  
Daniele Migliorini ◽  
Laura Baldassarri ◽  
Ilaria Boschi ◽  
...  
Keyword(s):  
Genetic Markers ◽  
Cannabis Sativa ◽  
Fiber Type ◽  
Nucleotide Polymorphisms ◽  
Loss Of Function ◽  
Single Nucleotide ◽  
Identification Process ◽  
Cannabidiolic Acid ◽  
Drug Type ◽  
Industrial Hemp

Genetic markers can be used in seeds and in plants to distinguish drug-type from fiber-type Cannabis Sativa L. varieties even at early stages, including pre-germination when cannabinoids are not accumulated yet. With this aim, this paper reports sequencing results for tetrahydrocannabinolic acid synthase (THCAS) and cannabidiolic acid synthase (CBDAS) genes from 21 C. sativa L. varieties. Taking into account that THCAS- and CBDAS-derived enzymes compete for the same substrate, the novelty of this work relies in the identification of markers based on both THCAS and CBDAS rather than THCAS alone. Notably, in our panel, we achieved an adequate degree of discrimination (AUC 100%) between drug-type and fiber-type cannabis samples. Our sequencing approach allowed identifying multiple genetic markers (single-nucleotide polymorphisms—SNPs—and a deletion/insertion) that effectively discriminate between the two subgroups of cannabis, namely fiber type vs. drug type. We identified four functional SNPs that are likely to induce decreased THCAS activity in the fiber-type cannabis plants. We also report the finding on a deletion in the CBDAS gene sequence that produces a truncated protein, possibly resulting in loss of function of the enzyme in the drug-type varieties. Chemical analyses for the actual concentration of cannabinoids confirmed the identification of drug-type rather than fiber-type genotypes. Genetic markers permit an early identification process for forensic applications while simplifying the procedures related to detection of therapeutic or industrial hemp.

scholarly journals In vitro propagation of Citrullus lanatus Thumb. from nodal explants culture

1970 ◽  
Vol 8 (2) ◽  
pp. 203-206 ◽  
Author(s):  
MM Khatun ◽  
MS Hossain ◽  
MA Haque ◽  
M Khalekuzzaman
Keyword(s):  
Citrullus Lanatus ◽  
In Vitro Regeneration ◽  
Shoot Proliferation ◽  
Multiple Shoot ◽  
Nodal Explants ◽  
Root Induction ◽  
Ms Medium ◽  
Nodal Explant ◽  
Grown Plant

A standard protocol was established for rapid in vitro propagation of watermelon (Citrullus lanatus Thumb.) from nodal explants of field grown plant. Multiple shoot proliferation was achieved from nodal explants on MS medium supplemented with 1.0 mg/l BAP + 0.2 mg/l NAA within 30 days of inoculation. The elongation of shoots was obtained on the same medium. Highest percentage of root induction was achieved on MS medium supplement with 1.0 mg/l IBA within 25 days of culture. Well rooted plantlets were transferred to small pots and after proper acclimatization the plantlets were transplanted in the field condition, where 80% plantlets were survived and grew successfully. Keywords: In vitro regeneration; Nodal explant; Citrullus lanatus DOI: 10.3329/jbau.v8i2.7926 J. Bangladesh Agril. Univ. 8(2): 203-206, 2010  

Sign in / Sign up

Export Citation Format

Share Document