scholarly journals Enhanced Production of β-Caryophyllene by Farnesyl Diphosphate Precursor-Treated Callus and Hairy Root Cultures of Artemisia vulgaris L.

2021 ◽  
Vol 12 ◽  
Author(s):  
Sundararajan Balasubramani ◽  
B. D. Ranjitha Kumari ◽  
Anil Kumar Moola ◽  
D. Sathish ◽  
G. Prem Kumar ◽  
...  
Keyword(s):  
Hairy Root ◽  
Leaf Explants ◽  
Farnesyl Diphosphate ◽  
Gas Chromatography Mass Spectrometry ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Artemisia Vulgaris ◽  
Enhanced Production ◽  
Wide Range ◽  
Half Strength

Artemisia vulgaris L. produces a wide range of valuable secondary metabolites. The aim of the present study is to determine the effects of various concentrations of farnesyl diphosphate (FDP) on β-caryophyllene content in both callus and hairy root (HR) cultures regeneration from leaf explants of A. vulgaris L. Murashige and Skoog (MS) medium supplemented with various concentrations of 2,4-dichlorophenoxyacetic acid (2,4D; 4–13 μM), α-naphthaleneacetic acid (NAA; 5–16 μM), and FDP (1 and 3 μM) was used for callus induction and HR regeneration from leaf explants of A. vulgaris L. In this study, precursor-treated (2,4D 13.5 μM + FDP 3 μM) callus displayed the highest biomass fresh weight (FW)/dry weight (DW): 46/25 g, followed by NAA 10.7 μM + FDP 3 μM with FW/DW: 50/28 g. Two different Agrobacterium rhizogenes strains (A4 and R1000) were evaluated for HR induction. The biomass of HRs induced using half-strength MS + B5 vitamins with 3 μM FDP was FW/DW: 40/20 g and FW/DW: 41/19 g, respectively. To determine β-caryophyllene accumulation, we have isolated the essential oil from FDP-treated calli and HRs and quantified β-caryophyllene using gas chromatography–mass spectrometry (GC–MS). The highest production of β-caryophyllene was noticed in HR cultures induced using A4 and R1000 strains on half-strength MS medium containing 3 μM FDP, which produced 2.92 and 2.80 mg/ml β-caryophyllene, respectively. The optimized protocol can be used commercially by scaling up the production of a β-caryophyllene compound in a short span of time.

scholarly journals IN VITRO REGENERATION AND MASS PROPAGATION OF AZIMA TETRACANTHA [LAM.] FROM THE LEAF EXPLANTS THROUGH CALLUS CULTURE

2017 ◽  
Vol 4 (2) ◽  
pp. 52-56
Author(s):  
Mallika Devi T
Keyword(s):  
Callus Induction ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Ms Medium ◽  
Apical Leaf ◽  
Half Strength ◽  
Regenerated Plantlets ◽  
Azima Tetracantha

In the present study the protocol for callus induction and regeneration in Azima tetracantha has been developed in culture medium. The young apical leaf explants were used for callus induction on MS medium containing BAP and NAA at 1.0 and 0.4mgl-1 respectively showed maximum callus induction (73%). The amount of callus responded for shoot formation (74%) was obtained in the MS medium containing BAP (1.5 mgl-1) and NAA (0.3mgl-1).The elongated shoots were rooted on half strength medium supplemented with IBA (1.5 mgl-1) and Kn (0.4 mgl-1) for shoots rooted. Regenerated plantlets were successfully acclimatized and hardened off inside the culture and then transferred to green house with better survival rate.

scholarly journals Establishment of a Rapid and Efficient Micropropagation System for Succulent Plant Haworthia turgida Haw.

HortScience ◽  
2017 ◽  
Vol 52 (9) ◽  
pp. 1278-1282 ◽  
Author(s):  
Boling Liu ◽  
Hongzhou Fang ◽  
Chaorong Meng ◽  
Ming Chen ◽  
Qingdong Chai ◽  
...  
Keyword(s):  
Callus Induction ◽  
Adventitious Shoots ◽  
Germplasm Conservation ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Induction Medium ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Succulent Plant

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.

scholarly journals Induction of Callogenesis, Organogenesis, and Embryogenesis in Non-Meristematic Explants of Bleeding Heart and Evaluation of Chemical Diversity of Key Metabolites from Callus

2020 ◽  
Vol 21 (16) ◽  
pp. 5826
Author(s):  
Dariusz Kulus ◽  
Alicja Tymoszuk
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Chemical Diversity ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Explant Type ◽  
Leaf Petiole ◽  
Primary And Secondary Metabolites ◽  
Culture Systems ◽  
Lamprocapnos Spectabilis

Lamprocapnos spectabilis (L.) Fukuhara is a perennial plant species valued in the horticultural, cosmetic, and pharmaceutical markets. To date, however, there were no studies on tissue culture systems in this species when adjusted from non-meristematic explants. The aim of this study is to induce callogenesis, organogenesis, and somatic embryogenesis in non-meristematic explants of Lamprocapnos spectabilis ‘Alba’ cultured in various media and to analyze the chemical diversity of the produced callus. Leaf, petiole, and internode explants were cultured on the modified Murashige and Skoog (MS) medium fortified with various combinations and concentrations of 6-benzyladenine (BA), indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorphenoxyacetic acid (2,4-D), and picloram (PIC). After 10 weeks of culturing, the morphogenetic response of explants was evaluated and the concentration of chlorophylls, carotenoids, anthocyanins, and polyphenols in callus was analyzed. There was no influence of explant type on the callogenesis efficiency (62.1–65.3%). The highest fresh weight of callus was produced on leaf explants in the presence of 2,4-D or PIC. In contrast, the highest share of dry weight was found in internode-derived calli and cultured on IAA-supplemented medium (up to 30.8%). Only 2.5% of all explants regenerated adventitious shoots, while rhizogenesis was reported in 4.5% of explants. Somatic embryos were produced indirectly by 0% to 100% of explants, depending on the culture medium and explant type. The highest mean number of embryos (11.4 per explant) was found on petioles cultured in the MS medium with 0.5 mg·L−1 BA and 1.0 mg·L−1 PIC. Calli cultured in media with NAA usually contained a higher content of primary and secondary metabolites. There was also a significant impact of explant type on the content of anthocyanins, polyphenols, and carotenoids in callus. Further studies should focus on the elicitation of metabolites production in callus culture systems of the bleeding heart.

scholarly journals Micropropagation and Quantification of Bioactive Compounds in Mertensia maritima (L.) Gray

2019 ◽  
Vol 20 (9) ◽  
pp. 2141 ◽  
Author(s):  
Han Yong Park ◽  
Doo Hwan Kim ◽  
Ramesh Kumar Saini ◽  
Judy Gopal ◽  
Young-Soo Keum ◽  
...  
Keyword(s):  
Linolenic Acid ◽  
Large Scale ◽  
Shoot Tip ◽  
Mass Spectrometry Analysis ◽  
Gas Chromatography Mass Spectrometry ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Root Induction ◽  
Ms Medium ◽  
Leaf Tissues

The goal of this study was to establish an efficient protocol for the large-scale propagation of Mertensia maritima (L.) Gray, and evaluate the carotenoid, fatty acid, and tocopherol contents in the leaves of in vitro regenerated shoots. Surface-disinfected node and shoot tip explants were placed on semisolid Murashige and Skoog (MS) medium with 0–16 µM N6-benzyladenine (BA), kinetin, (KN), and thidiazuron (TDZ) alone, or in combination with, 1 or 2 µM α-naphthaleneacetic acid (NAA). Of the three different cytokinins employed, TDZ elicited the best results for axillary shoot proliferation. A maximum frequency of shoot initiation above 84%, with a mean of 8.9 and 4.8 shoots per node and shoot tip, respectively, was achieved on the culture medium supplemented with 4 µM TDZ. A combination of TDZ + NAA significantly increased the percentage of multiple shoot formation and number of shoots per explant. The best shoot induction response occurred on MS medium with 4 µM TDZ and 1 µM NAA. On this medium, the node (93.8%) and shoot tip (95.9%) explants produced an average of 17.7 and 8.6 shoots, respectively. The highest root induction frequency (97.4%) and number of roots per shoot (25.4), as well as the greatest root length (4.2 cm), were obtained on half-strength MS medium supplemented with 4 µM indole-3-butyric acid (IBA). The presence of six carotenoids and α-tocopherol in the leaf tissues of M. maritima was confirmed by HPLC. Gas chromatography-mass spectrometry analysis confirmed the presence of 10 fatty acids, including γ-linolenic acid and stearidonic acid in the leaf tissues of M. maritima. All-E-lutein (18.49 μg g−1 fresh weight, FW), α-tocopherol (3.82 μg g−1 FW) and α-linolenic acid (30.37%) were found to be the significant compounds in M. maritima. For the first time, a successful protocol has been established for the mass propagation of M. maritima with promising prospects for harnessing its bioactive reserves.

scholarly journals Transformation of Nicotiana alata Link and Otto. with an Autoregulatory Senescence-inhibitor Gene Construct

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 617c-617
Author(s):  
Kenneth R. Schroeder ◽  
Dennis P. Stimart
Keyword(s):  
Agrobacterium Tumefaciens ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Nicotiana Alata ◽  
Ms Medium ◽  
Gene Encoding ◽  
Half Strength ◽  
Cytokinin Synthesis ◽  
Senesced Leaves

Leaf explants of Nicotiana alata Link and Otto. were surface disinfested and cultured on Murashige and Skoog (MS) medium containing 2.66 μm N6-benzyladenine (BA) to promote shoot proliferation. After 5 weeks, proliferated shoots were removed and remaining callus saved. Callus was inoculated with Agrobacterium tumefaciens encoding a senescence-specific promoter SAG12 cloned from Arabidopsis thaliana fused to a Agrobacterium tumefaciens gene encoding isopentenyl transferase which catalyzes cytokinin synthesis. Following inoculation, the callus was cocultivated for 6 days on BA medium. Selection for transgenics was done on BA medium plus 100 mg Kanamycin and 400 mg Ticarcillin (antibiotics) per liter. Proliferating shoots were rooted on MS medium containing antibiotics. Rooted cuttings were transplanted to soil, acclimated and flowered in the greenhouse. Transgenics were outcrossed to a commercial N. alata hybrid. Seed was germinated in vitro on half-strength MS medium plus antibiotics. Segregation of transgenics to nontransgenics was 1:1. Evaluation of leaf senescence on 5-month-old plants showed 2 to 14 times fewer senesced leaves on the transgenic than the nontransgenic plants.

scholarly journals Resveratrol Production from Hairy Root Cultures of Scutellaria baicalensis

2013 ◽  
Vol 8 (5) ◽  
pp. 1934578X1300800
Author(s):  
Sang-Won Lee ◽  
Young Seon Kim ◽  
Md. Romij Uddin ◽  
Do Yeon Kwon ◽  
Yeon Bok Kim ◽  
...  
Keyword(s):  
Hairy Root ◽  
Scutellaria Baicalensis ◽  
Growth Media ◽  
Hairy Root Cultures ◽  
Ms Medium ◽  
Root Cultures ◽  
Full Strength ◽  
Alternative Approach ◽  
Half Strength ◽  
The Media

The levels of resveratrol produced by hairy root cultures of Scutellaria baicalensis were investigated using different media of varying strengths and in the presence of various concentrations of auxins. The levels of resveratrol were higher when the hairy root cultures were maintained in full-strength Murashige and Skoog (MS) medium when compared with other growth media. The cultures grown in full-strength MS medium produced 2.5-fold higher resveratrol than those grown in half-strength B5 medium—the lowest resveratrol-producing medium. The levels of resveratrol varied significantly when cultures were grown in full-strength MS with varying concentrations of auxins. Supplementation of the media with the auxin indole acetic acid (IAA) at 0.1 mg/L produced the highest accumulation of resveratrol. Our findings reveal a valuable alternative approach for the production of resveratrol from S. baicalensis.

Development of an elicitation strategy on enhanced accumulation of oleanolic acid in suspension cultures of Ocimum tenuiflorum L.

2021 ◽  
Author(s):  
Swati Sharan ◽  
Neera Bhalla Sarin ◽  
Kunal Mukhopadhyay
Keyword(s):  
Oleanolic Acid ◽  
Biomass Production ◽  
Cell Cultures ◽  
Leaf Explants ◽  
Suspension Cultures ◽  
Suspension Cell ◽  
Naphthaleneacetic Acid ◽  
Enhanced Production ◽  
Ocimum Tenuiflorum ◽  
Suspension Cell Cultures

Abstract Ocimum tenuiflorum Linn. is an important aromatic medicinal plant which produces several secondary metabolites responsible for diverse pharmacological activities. The present study focuses on enhanced production of biomass as well as oleanolic acid (OA), an anticancer and antioxidant compound, in suspension cell cultures of O. tenuiflorum upon elicitation with different elicitors. Leaf explants derived friable calli were inoculated intol iquid Murashige and Skoog (MS) media containing plant growth regulators [0.25 mg/L of α-naphthaleneacetic acid (NAA) and 0.5 mg/L of 6-benzyl amino purine (BAP)] for the establishment of suspension cultures. Influence of several factors such as, age of the suspension cultures, different concentrations, and exposure times of various elicitors such as yeast extract (YE), methyl jasmonate (MeJ) and salicylic acid (SA) respectively were analysed for cell biomass production and accumulation of OA during this study. Among the elicitors tested, YE at 50 mg/L was found to be the most efficient in terms of increased biomass production and accumulation of OA in the cultures. The highest increase in OA production (13.16-fold in elicited cultures compared to untreated cultures) was noted on 17-day-old suspension cultures when exposed to 50 mg/L of YE for four days. Enhancement of 2.72-fold in OA content was also recorded in 17-day-old cultures when treated with MeJ (60 mg/L) during two days of exposure. SA was not efficient in inducing accumulation of OA in 17- and 22-day-old suspension cultures at any concentration and exposure time. Furthermore, it was observed that the effect of different elicitors on biomass production and OA content depended on concentration and the duration of exposure times. Therefore, utilization of elicitation method could be a promising tool to enhance cell growth and OA accumulation in the suspension cell cultures of O. tenuiflorum.

scholarly journals Propagation of goldenrod (Solidago canadensis L.) from leaf and nodal explants

2012 ◽  
Vol 81 (1) ◽  
pp. 53-60 ◽  
Author(s):  
Jun Li ◽  
Ye Kang ◽  
Sheng Qiang ◽  
Gary Peng
Keyword(s):  
Invasive Plant ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Nodal Segments ◽  
Naphthalene Acetic Acid ◽  
Axillary Shoot ◽  
Solidago Canadensis ◽  
Ms Medium ◽  
Regeneration System ◽  
Half Strength

Goldenrod (<em>Solidago canadensis </em>L.) is an invasive plant species in many countries except North America but a cut-flower species worldwide. There is a need to generate and propagate goldenrod clones efficiently for research and commercial purposes. A callus induction and plantlet regeneration system was developed by studying the influence of explant type and different concentrations of plant growth regulators. The highest callus production from leaf segments was obtained on Murashige and Skoog’s medium (MS medium) supplemented with 1.0 mg/L naphthalene acetic acid (NAA) and 1.0 mg/L 6-benzylaminopurine (BA). Adventitious shoots could be regenerated directly from leaf explants without an intermediate callus phase with the highest shoot induction percentage of 87.2%. The largest number of adventitious shoots per leaf explant (3.2) was obtained on MS medium supplemented with 0.4 mg/L NAA and 2.0 mg/L BA. MS medium supplemented with 0.1 mg/L NAA and 1.0 mg/L BA was the best medium for axillary shoot regeneration from nodal segments. The highest root number and longest roots occurred on half-strength MS without the addition of any growth regulator. Rooted plantlets were then transferred to a soil-based growth medium, placed in a greenhouse, and acclimatized with 100% success. All surviving plants grew normally without showing any morphological varia&shy;tion when compared to those grow from seed. This regeneration protocol may be used to produce certain biotypes of goldenrod suitable for genetic transformation rapid propagation of goldenrod for commercial purposes or for screening fungi and toxins as potential biocontrol agents against this weed.

scholarly journals High Efficiency Direct Shoot Organogenesis from Leaf Segments ofAerva lanata(L.) Juss. Ex Schult by Using Thidiazuron

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
K. Varutharaju ◽  
C. Soundar Raju ◽  
C. Thilip ◽  
A. Aslam ◽  
A. Shajahan
Keyword(s):  
Shoot Organogenesis ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Naphthaleneacetic Acid ◽  
Scale Production ◽  
Ms Medium ◽  
Leaf Segments ◽  
Direct Shoot Organogenesis ◽  
Direct Shoot

An efficient protocol for direct shoot organogenesis has been developed for the medicinal plantAerva lanata(L.) Juss. ex Schult. Regeneration was achieved from leaf segments of 20 days oldin vitroplantlets raised on Murashige and Skoog (MS) medium containing 0.25–2.0 mg L−1thiadiazuron (TDZ), 3% sucrose, and 0.8% agar. After 21 days of culture incubation, maximum number of shoot organogenesis (23.6 ± 0.16) was obtained on medium containing 1.0 mg L−1TDZ. The shoots were able to producein vitroflowers on medium containing 1.0 mg L−1TDZ in combination with 0.25–0.5 mg L−1  α-naphthaleneacetic acid (NAA). Histological observation showed that the epidermal cells of the leaf explants exhibited continuous cell division led to formation of numerous dome shaped meristematic protrusions and subsequently developed into adventitious shoots. Upon transfer of shootlets to half strength MS medium containing 1.0 mg L−1indole-3-butyric acid (IBA), around 86% of the regenerated shoots formed roots and plantlets. Rooted plants were hardened and successfully established in the soil at the survival rate of 92%. The regeneration protocol developed in this study provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for a large scale production of its medicinally active compounds and genetic transformations for further improvement.

scholarly journals In vitro Propagation of Solanecio biafrae and Determination of Genetic Stability of Plantlets Using RAPD and ISSR Markers

2016 ◽  
Vol 24 (1) ◽  
pp. 29-36 ◽  
Author(s):  
Jelili Opabode ◽  
Oluyemisi Akinyemiju
Keyword(s):  
Random Amplified Polymorphic Dna ◽  
Issr Markers ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Germplasm Exchange ◽  
Indole Butyric Acid ◽  
Half Strength ◽  
Micropropagated Plants

Abstract An efficient and reproducible micropropagation protocol of Solanecio biafrae (Oliv. & Hiern) C. Jeffrey has been developed from nodal stem segments. Shoot development was obtained on Murashige and Skoog (MS) medium supplemented with benzylaminopurine (BAP) alone and in combination with zeatin and 1-naphthaleneacetic acid (NAA). Elongated shoots were rooted in the presence of zeatin or 3-indole-butyric acid (IBA) alone or in combinations. The highest number of explants forming shoots (100%) as well as the highest number of shoots per explant (3.4) and the longest shoots (22 mm) were recorded on medium containing 4.0 mg·dm−3 BAP, 2.0 mg·dm−3 NAA, and 1.0 mg·dm−3 zeatin. About 76% of shoots formed roots on half-strength MS medium free of plant growth regulators. The best root formation (approximately 88%) was recorded on the medium containing 1.0-1.5 mg·dm−3 IBA. The micropropagated shoots with well-developed roots were efficiently acclimatized under greenhouse conditions. The random amplified polymorphic DNA (RAPD) and intersimple sequence repeat (ISSR) amplification products were monomorphic in micropropagated plants and similar to those of mother plant showing their genetic uniformity. This is the first report of micropropagation of S. biafrae, which will facilitate in vitro mass propagation, conservation, and germplasm exchange of this endangered African vegetable.

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