Expression Analysis of Phenylpropanoid Pathway Genes and Metabolomic Analysis of Phenylpropanoid Compounds in Adventitious, Hairy, and Seedling Roots of Tartary Buckwheat

Tartary buckwheat (Fagopyrum tataricum) is an important crop that belongs to the Polygonaceae family, whose roots have received considerable attention due to the presence of compounds with high nutritional and medicinal value. In this study, we aimed to develop an efficient protocol for the culture of adventitious (ARs) and hairy (HRs) roots on a half-strength Schenk and Hildebrandt (SH) medium containing different concentrations of the auxins, α-naphthaleneacetic acid (NAA), indole-3-butyric acid (IBA), and indole-3-acetic acid (IAA). The highest percentage of root induction (91.67%) was achieved with 0.5 mg/L IAA, whereas the greatest number of roots was found in 1 mg/L IAA. In contrast, 0.1 mg/L IBA returned the longest roots. As expected, HRs were obtained from in vitro leaf explants infected with Agrobacterium rhizogenes R1000. Quantitative real-time polymerase chain reaction (qRT-PCR) analysis of 11 phenolic pathway genes revealed that five genes (FtPAL, FtC3H, FtHQT, FtCHS, and FtANS) were highly expressed in HRs, whereas only four (FtC4H, FtFLS2, FtDFR, and FtANR), and three (Ft4CL, FtCHI, and FtF3H) were recognized in the ARs and seedling roots (SRs), respectively. HPLC analysis of phenolic compounds in different root cultures showed that the majority of the phenolic compounds (both individual and total) were significantly accumulated in the HRs. Principal component analysis (PCA) identified differences among the three root types, whereby HRs were separated from ARs and SRs based on the amount of phenolic compounds present. Analysis of the metabolic pathway revealed that among the identified metabolites, the 3, 2, and 1 pathways were associated with flavonoid, flavone and flavonol, and phenylpropanoid biosynthesis, respectively. Hierarchical clustering analysis and the heat map showed that the different root cultures presented unique metabolites.

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In vitro shoot regeneration of boldo from leaf explants

Ciência Rural ◽

10.1590/s0103-84782010005000173 ◽

2010 ◽

Vol 40 (10) ◽

pp. 2210-2213

Author(s):

Monalize Salete Mota ◽

Juliana de Magalhães Bandeira ◽

Eugenia Jacira Bolacel Braga ◽

Valmor João Bianchi ◽

José Antonio Peters

Keyword(s):

Shoot Regeneration ◽

Leaf Explants ◽

Shoot Development ◽

High Potential ◽

Naphthaleneacetic Acid ◽

Regeneration System ◽

Bud Induction ◽

Molecular Studies ◽

In Vitro Shoot Regeneration

A shoot regeneration system for Plectranthus neochilus was studied from leaf explants. Leaves developed under in vitro conditions were cultured on Wood Plant Medium supplemented with 0.2mg dm-3 α-naphthaleneacetic acid (NAA) and different 6-benzilaminopurine (BAP) or thidiazuron (TDZ) concentrations (0, 1.5, 3.0, 4.5 and 6.0mg dm-3). An increase in percentage of responsive explants (85.3%) and in the number of shoots developed per explant (3.2) was observed when the explants were treated with 5.3 and 4.7mg dm-3 BAP, respectively. The leaf explants cultured on media supplemented with TDZ became vitreous and did not form buds. The regeneration system used is efficient for boldo bud induction and shoot development, showing high potential for advanced cellular and molecular studies.

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In vitro regeneration protocol of Rauvolfia serpentina L.

Bangladesh Journal of Scientific and Industrial Research ◽

10.3329/bjsir.v53i2.36674 ◽

2018 ◽

Vol 53 (2) ◽

pp. 133-138 ◽

Cited By ~ 1

Author(s):

S Khan ◽

TA Banu ◽

S Akter ◽

B Goswami ◽

M Islam ◽

...

Keyword(s):

In Vitro Regeneration ◽

Leaf Explants ◽

Multiple Shoot ◽

Nodal Segments ◽

Root Induction ◽

Ms Medium ◽

Regeneration System ◽

Rauvolfia Serpentina ◽

Number Of Shoots

An efficient in vitro regeneration system was developed for Rauvolfia serpentina L. through direct and indirect organogenesis from nodal and leaf explants. Among the different growth regulators, MS medium supplemented with 2.0 mg/l BAP, 0.5mg/l IAA and 0.02mg/l NAA found best for the multiple shoot formation from nodal segments. In this combination 98% explants produced multiple shoots and the average number of shoots per explants is 13∙4. The frequency of callus induction and multiple shoot induction from leaves was highest 88% in MS medium supplemented with 2.0 mg/l BAP, where mean number of shoots/explants was 12.5. The highest frequency of root induction (80%) and mean number of roots/plantlets (10) were obtained on half strength of MS medium containing 0.2 mg/l IBA. The rooted plantlets were transferred for hardening following acclimatization and finally were successfully established in the field.Bangladesh J. Sci. Ind. Res.53(2), 133-138, 2018

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

Botanica Orientalis Journal of Plant Science ◽

10.3126/botor.v6i0.2918 ◽

2010 ◽

Vol 6 ◽

pp. 103-105 ◽

Cited By ~ 5

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

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

HortScience ◽

10.21273/hortsci12056-17 ◽

2017 ◽

Vol 52 (9) ◽

pp. 1278-1282 ◽

Cited By ~ 4

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.

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In vitro Regeneration and Agrobacterium-mediated Genetic Transformation of Tomato (Lycopersicon esculentum Mill.)

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i1.5004 ◽

1970 ◽

Vol 19 (1) ◽

pp. 101-111 ◽

Cited By ~ 1

Author(s):

Rakha Hari Sarker ◽

Khaleda Islam ◽

M.I. Hoque

Keyword(s):

Lycopersicon Esculentum ◽

Genetic Transformation ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Multiple Shoots ◽

Multiple Shoot ◽

Root Induction ◽

Cotyledonary Leaf ◽

Lycopersicon Esculentum Mill

Agrobacterium-mediated genetic transformation system has been developed for two tomato (Lycopersicon esculentum Mill.) varieties, namely Pusa Ruby (PR) and BARI Tomato-3 (BT-3). Prior to the establishment of transformation protocol cotyledonary leaf explants from the two varieties were cultured to obtain genotype independent in vitro regeneration. Healthy multiple shoot regeneration was obtained from the cut ends of cotyledonary leaf segments for both the varieties on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. The maximum root induction from the regenerated shoots was achieved on half the strength of MS medium supplemented with 0.2 mg/l IAA. The in vitro grown plantlets were successfully transplanted into soil where they flowered and produced fruits identical to those developed by control plants. Transformation ability of cotyledonary leaf explants was tested with Agrobacterium tumefaciens strain LBA4404 harboring binary plasmid pBI121, containing GUS and npt II genes. Transformed cotyledonary leaf explants were found to produce multiple shoots on MS containing 1.0 mg/l BAP and 0.1 mg/l IAA. Selection of the transformed shoots was carried out by gradually increasing the concentration of kanamycin to 200 mg/l since kanamycin resistant gene was used for transformation experiments. Shoots that survived under selection pressure were subjected to rooting. Transformed rooted plantlets were transferred to soil. Stable expression of GUS gene was detected in the various tissues from putatively transformed plantlets using GUS histochemical assay. Key words: In vitro regeneration, transformation, tomato D.O.I. 10.3329/ptcb.v19i1.5004 Plant Tissue Cult. & Biotech. 19(1): 101-111, 2009 (June)

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Honokiol and Magnolol Production by in vitro Micropropagated Plants of Magnolia dealbata, an Endangered Endemic Mexican Species

Natural Product Communications ◽

10.1177/1934578x1000500213 ◽

2010 ◽

Vol 5 (2) ◽

pp. 1934578X1000500 ◽

Cited By ~ 4

Author(s):

Fabiola Domínguez ◽

Marco Chávez ◽

María Luisa Garduño-Ramírez ◽

Víctor M. Chávez-Avila ◽

Martín Mata ◽

...

Keyword(s):

Growth Medium ◽

Leaf Explants ◽

Wild Plants ◽

Root Induction ◽

Ms Medium ◽

Plant Materials ◽

Detection Techniques ◽

Magnolia Dealbata ◽

Selection Of

An efficient protocol for the in vitro propagation of Magnolia dealbata Zucc., an important medicinal plant that is the source of the anxiolytic and anticancer compounds honokiol and magnolol, was established. This plant is wild-crafted, and conservationists have expressed concerns with regard to the sustainability of production. In the present work, two factors were found to be of importance for the regeneration of M. dealbata and the production of honokiol and magnolol. These factors were the type of explants and the combination and concentration of plant-growth regulators. Green, compact, nodular organogenic callus was obtained from leaf explants in a medium fortified with Murashige and Skoog salts and supplemented with 1.5 mg/L 2,4-dicholorophenoxyacetic acid and 1.5 mg/L kinetin. Shoot multiplication from callus cultures was achieved in the Murashige and Skoog (MS) medium with 1.5 mg/L thidiazuron (TDZ). Phenol secretion was controlled by the addition of 250 mg/L of activated charcoal. For rooting, shoots were transferred to MS medium supplemented with several auxins. After root induction, the plants were hardened in earthen pots containing sand, soil, and vermiculite. The contents of honokiol (HK) and magnolol (MG) were determined in different plant materials by high-performance liquid chromatography-diode-array detection techniques. This analysis revealed that the honokiol and magnolol content in aerial and underground parts of micropropagated M. dealbata were higher than that observed in wild plants (both 6 months old). Our results suggest that conservation of M. dealbata is possible by means of in vitro multiplication of leaf-derived callus. The usefulness of M. dealbata regeneration and production of HK and MG may be attributed to the proper selection of explant sourcing and identification of the correct growth medium to support adequate growth. This careful selection of explants and growth medium leads to a very useful source of plant material for pharmacological and phytomedicinal screening applications and, above all, would safeguard this plant species from the threat of extinction.

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Total Polyphenol Content, in vitro Antifungal and Antioxidant Activities of Callus Cultures from Inula crithmoides

Natural Product Communications ◽

10.1177/1934578x1300801122 ◽

2013 ◽

Vol 8 (11) ◽

pp. 1934578X1300801 ◽

Cited By ~ 2

Author(s):

Anahi Bucchini ◽

Laura Giamperi ◽

Donata Ricci

Keyword(s):

Methanol Extract ◽

Antioxidant Activities ◽

Leaf Explants ◽

Basal Medium ◽

Alternaria Solani ◽

Antimicrobial Activities ◽

Callus Cultures ◽

Naphthaleneacetic Acid ◽

Stable Free Radical

This is the first report on the antioxidant and antifungal activities of callus cultures from Inula crithmoides L. (Asteraceae). Callus cultures were initiated from leaf sections, on initial culture MS basal medium supplemented with various concentrations of 2,4-D (2,4-dichlorophenoxyacetic acid), NAA (1-naphthaleneacetic acid) and IBA (indole-3-butyric acid) and a 72% survival was achieved. Significant differences between the various auxins used as phytohormones on callus growth were found. Maximum callusing was noticed on the leaf explants grown on MS basal medium supplemented with 1 mgL–1 2,4-D. Subsequently the antioxidant and antimicrobial activities of the methanol extract from calli were investigated. Antioxidant studies suggested that the methanol extracts of dark-grown and light-grown callus were able to reduce the stable free radical 2,2-diphenyl-1-picrilhydrazyl (DPPH). In the inhibition against lipid peroxidation, extracts of dark-grown callus showed the strongest effect with IC50 values better than those of the standards. The methanol extract of callus cultures had significant antifungal activity only against two of the fungi tested: Alternaria solani and Phytophthora cryptogea. Against all the other tested fungi, the I. crithmoides calli extracts showed fungistatic activity.

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REGENERATION OF TREMA ORIENTALIS (BLUME) LINN.: EFFECT OF GROWTH REGULATORS, CULTURE CONDITIONS, AND AGE AND SOURCE OF EXPLANTS

Israel Journal of Plant Sciences ◽

10.1080/07929978.1995.10676625 ◽

1995 ◽

Vol 43 (4) ◽

pp. 391-395 ◽

Cited By ~ 1

Author(s):

G.R. Rout ◽

S. Samantaray ◽

P. Das

Keyword(s):

Growth Regulators ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Mature Trees ◽

Regeneration Rate ◽

Bud Regeneration ◽

Shoot Bud ◽

Shoot Bud Regeneration ◽

Trema Orientalis

Optimal conditions for high frequency shoot bud regeneration from leaf callus of Trema orientalis (Blume) Linn. were studied. The regeneration rate was controlled by the growth regulators, the age and the source of the explants, and the illumination conditions. Irrespective of illumination conditions, shoot bud regeneration was achieved only in media containing benzyladenine (BA) + α-naphthaleneacetic acid (NAA) combinations, with the best results being obtained in the presence of 2.5 mg/1 BA and 0.25–0.5 mg/1 NAA. The morphogenic response was less frequent in the calluses derived from leaf explants of the mature trees compared to those of the in vitro-grown seedlings. The rate of shoot bud regeneration was more pronounced in the cultures maintained for 4 weeks in the light (16-h photoperiod) than the cultures incubated in the dark. Regenerated shoots were rooted on the medium containing 1/2 strength basal Murashige and Skoog (MS) salts supplemented with 0.01 mg/1 NAA or indole-3-butyric acid (IBA). The rooted plantlets were established in the greenhouse.

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CALLUS INDUCTION FROM LEAF EXPLANT OF FICUS DELTOIDEA VARKUNSTLERI

Science Heritage Journal ◽

10.26480/gws.01.2020.06.08 ◽

2020 ◽

Vol 4 (1) ◽

pp. 06-08

Author(s):

Hafizuddin Sa’adan ◽

Zarina Zainuddin

Keyword(s):

Blood Circulation ◽

Leaf Explants ◽

Mercury Chloride ◽

Root Induction ◽

Ficus Deltoidea ◽

Surface Sterilization ◽

Pre Treatment ◽

In Vitro Clonal Propagation ◽

Herbal Plant

Ficus deltoidea or commonly known as ‘mas cotek’ is a herbal plant indigenous to Southeast Asia including Malaysia and Indonesia. This plant is popular for its medicinal values such as improve blood circulation, regain energy and enhance fertility naturally for both men and women. The main objective of this study is to develop in vitro clonal propagation method for rapid production of F. deltoidea using different concentrations of benzyl aminopurine (BAP) through shoot induction and multiplication, rooting and subsequent establishment in soil following acclimatization. Surface sterilization of the leaf explants was done using mercury chloride and ethanol as the disinfectants. Pre-treatment of the explants with carbendazim successfully reduced the occurrence of fungal contamination. At the end of the experiment, no shoot and root induction were observed but calli were successfully induced on MS medium containing 1.0, 1.5, 2.0, 2.5 and 3.0 mg/l BAP, with calli induced from 3.0 mg/l BAP were bigger and healthier. In short, the higher the concentration of BAP used, the higher tendency for the explant to induce callus.

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Isolated culture of A. reptance L., its’ morphological and growth features

BIO Web of Conferences ◽

10.1051/bioconf/20214001001 ◽

2021 ◽

Vol 40 ◽

pp. 01001

Author(s):

Elen Poghosyan ◽

Naira Sahakyan ◽

Margarit Petrosyan ◽

Irina Batlutskaya ◽

Karen Trchounian

Keyword(s):

Callus Culture ◽

Nutrient Medium ◽

Leaf Explants ◽

Shoot Formation ◽

Callus Cultures ◽

Soil Conditions ◽

Naphthaleneacetic Acid ◽

Micro Propagation ◽

2,4 Dichlorophenoxyacetic Acid

A growing demand for the ecologically pure products brings us for searching novel biotechnological approaches for plant cultivation. One of these approaches is the in vitro cultivation and further acclimatization of valuable plant species. The object of our investigation was Ajugareptance L. ornamental plant which possesses high metabolic activity. In vitro cultivation was carried out applying Murashige-Skoog nutrient medium and its modifications. Acclimatization of in vitro plants was implemented according Hazarika. In the presence of twice higher concentration of cytokinins over auxins and 0.2 mg/ml gibberellins callus culture was formed from the leaf explants. Callus tissue was formed in the presence of 0.2 mg/ml kinetin and 2 mg/ml indole-3-acetic acid which has denser structure than the first one. The shoot formation was observed on callus cultures growing on the same medium approximately after 5th passage. Callus culture growth was supported also by the adding of 2 mg/ml 2,4-dichlorophenoxyacetic acid. For the micropropagation, the already formed shoots were transferred to the nutrient medium which contains only 0.1 mg/ml 1-Naphthaleneacetic acid as a phytohormone. A. reptans culture has high regenerative ability and the micro-propagation index was 104 – 105. In vitro regenerated plants were successfully acclimatized to the soil conditions during two-week period.

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