scholarly journals Secure and Sustainable Sourcing of Plant Tissues for the Exhaustive Exploration of Their Chemodiversity

Molecules ◽  
2020 ◽  
Vol 25 (24) ◽  
pp. 5992
Author(s):  
Rhodin C. Joseph ◽  
Matheus Silva da Fonseca Diniz ◽  
Viviane Magno do Nascimento ◽  
Abraão de Jesus Barbosa Muribeca ◽  
Johan Carlos Costa Santiago ◽  
...  
Keyword(s):  
Leaf Explants ◽  
Chemical Diversity ◽  
Plant Tissues ◽  
Main Challenge ◽  
Plant Chemical ◽  
Leaf Tissues ◽  
Marker Compounds ◽  
Alternative Solutions ◽  
Sustainable Sourcing

The main challenge of plant chemical diversity exploration is how to develop tools to study exhaustively plant tissues. Their sustainable sourcing is a limitation as bioguided strategies and dereplication need quite large amounts of plant material. We examine if alternative solutions could overcome these difficulties by obtaining a secure, sustainable, and scalable source of tissues able to biosynthesize an array of metabolites. As this approach would be as independent of the botanical origin as possible, we chose eight plant species from different families. We applied a four steps culture establishment procedure, monitoring targeted compounds through mass spectrometry-based analytical methods. We also characterized the capacities of leaf explants in culture to produce diverse secondary metabolites. In vitro cultures were successfully established for six species with leaf explants still producing a diversity of compounds after the culture establishment procedure. Furthermore, explants from leaves of axenic plantlets were also analyzed. The detection of marker compounds was confirmed after six days in culture for all tested species. Our results show that the first stage of this approach aiming at easing exploration of plant chemodiversity was completed, and leaf tissues could offer an interesting alternative providing a constant source of natural compounds.

In vitro initiation of adventitious buds and its modification by high concentration of benzyladenine in leaf tissues of mulberry (Morus alba)

1981 ◽  
Vol 59 (1) ◽  
pp. 68-74 ◽  
Author(s):  
S. Oka ◽  
K. Ohyama
Keyword(s):  
Leaf Explants ◽  
Morus Alba ◽  
Adventitious Buds ◽  
High Concentration ◽  
Leaf Tissues ◽  
Murashige And Skoog's Medium ◽  
Murashige And Skoog’S Medium ◽  
Bud Initiation ◽  
The Way

Leaf explants of mulberry (Mortis alba L.) derived from aseptically grown shoots and seedlings were cultured on Murashige and Skoog's medium. Normal and abnormal leaves were grown by varying the concentration of benzyladenine. They differed in the way of forming adventitious buds. In normal leaves bud initiation occurred exclusively at the cut ends of midribs after removing petioles, while in abnormal ones buds formed at any region on midribs and petioles. Histological observations were made to study different patterns of bud initiation.

scholarly journals Plant Regeneration of Chokecherry (Prunus virginiana L.) from in vitro Leaf Tissues

2004 ◽  
Vol 22 (4) ◽  
pp. 225-228 ◽  
Author(s):  
Wenhao Dai ◽  
Victoria Jacques ◽  
James A. Walla ◽  
Zong-Ming Cheng
Keyword(s):  
Plant Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Regeneration System ◽  
Prunus Virginiana ◽  
Leaf Tissues ◽  
Half Strength ◽  
Eventual Death ◽  
Roots In Vitro

Abstract An effective plant regeneration system was developed for chokecherry (Prunus virginiana L.) by using in vitro leaf tissues. Adventitious shoots regenerated from in vitro leaf tissues only when cultured on Woody Plant Medium (WPM), but not on Murashige and Skoog medium, supplemented with benzyladenine (BA) or thidiazuron (TDZ). Three chokecherry clones (NN, 10, and 17) responded differently to types and concentrations of cytokinins, ranging from 16.7 to 91.7% leaf explants regenerating shoots. A mean of four shoots was produced from each explant, with the most shoots (> 10) from clone NN on media with 5–10 μM BA. Higher concentrations of TDZ (> 8 μM) caused serious vitrification and eventual death of newly induced shoots. Regenerated shoots (> 1.5 cm) produced roots in vitro in half strength MS medium or ex vitro in Cellular Rooting Sponge (CRS) rooting plugs with or without auxin (NAA or IBA) treatments. Rooting was affected by auxin, genotypes, and the rooting methods.

scholarly journals Interaction of Four Antagonistic Fungi with Botrytis aclada in Dead Onion Leaves: A Comparative Microscopic and Ultrastructural Study

1997 ◽  
Vol 87 (6) ◽  
pp. 634-642 ◽  
Author(s):  
J. Köhl ◽  
R. R. Bélanger ◽  
N. J. Fokkema
Keyword(s):  
Ultrastructural Level ◽  
Spore Production ◽  
Plant Tissues ◽  
Antagonistic Action ◽  
Biocontrol Activity ◽  
Leaf Tissues ◽  
Biocontrol Potential ◽  
In Vitro Interaction ◽  
Antagonistic Interactions

The colonization of dead onion leaves by Botrytis aclada and the fungal antagonists Aureobasidium pullulans, Chaetomium globosum, Glio-cladium catenulatum, and Ulocladium atrum and the interactions between B. aclada and each of the four antagonists were studied at the microscopic and ultrastructural level. This approach was used in an attempt to understand the colonization pattern of these fungi and the nature of the biocontrol activity of the antagonists that have shown a potential to suppress spore production of Botrytis spp. on necrotic plant tissues. When applied alone, B. aclada and U. atrum were found throughout the leaf tissues in high densities after an incubation period of 6 days at 18°C in a moist chamber. C. globosum and G. catenulatum colonized only the outer portions of the leaf, whereas A. pullulans appeared to be concentrated in the leaf stomata. When pathogen and antagonists were applied together, ultrastructural observations revealed that cells of B. aclada were plasmolyzed in the presence of G. catenulatum, suggesting a reaction to antifungal molecules. Antibiosis also seemed to be involved, albeit to a lesser extent, in the antagonistic interactions between B. aclada and A. pullulans or C. globosum. No evidence of direct parasitism was recorded. On the other hand, U. atrum appeared to completely exclude B. aclada from dead onion tissues when both fungi competed for the substrate. Ultrastructural observations of the in vitro interaction between the two fungi did not reveal parasitism or antibiosis by either fungus. Based on previous records of its biocontrol potential and observations of its colonizing properties, it appears that U. atrum can compete for and utilize necrotic tissues rapidly and extensively, thus, excluding competitors without any other antagonistic action.

scholarly journals Agrobacterium-mediated Transformation of Chokecherry (Prunus virginiana L.)

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 755B-755
Author(s):  
Wenhao Dai* ◽  
Christopher P. Johnson ◽  
Victoria A. Jacques ◽  
James A. Walla
Keyword(s):  
Transformation Efficiency ◽  
Leaf Explants ◽  
Wildlife Habitat ◽  
Infected Leaf ◽  
Small Tree ◽  
Prunus Virginiana ◽  
Gene Encoding ◽  
Leaf Tissues ◽  
Polymerase Chain

An Agrobacterium-mediated transformation system was developed for chokecherry (Prunus virginiana L.), one of the most popular native small tree or large shrub species for resource conservation and wildlife habitat in North America. Leaf tissues from in vitro plants previously maintained in MS medium with 2.5 μm BA were co-cultivated on woody plant medium (WPM) containing 10 μm BA and 200 μm acetosyringone with Agrobacterium tumefaciens strain EHA105 harboring the binary Ti plasmid pBI121 carrying the uid A gene encoding for β-glucuronidase (GUS) and the npt II gene encoding neomycin phosphotransferase II. Infected leaf explants were disinfected in sterile water and antibiotics and then transferred to WPM containing 10 μM BA and the antibiotics cefotaxime, carbenicillin, and kanamycin (CCK) for shoot regeneration at 25 °C with a 16-hour photoperiod. Agrobacterium concentration, pre-conditioning of explants, application of acetosyringone, infection time, and kanamycin tolerance of leaf tissues were evaluated for effects on transformation efficiency. Regeneration of chokecherry shoots on kanamycin-containing medium and screening by GUS histochemical assays showed that both the npt II and the uid A genes were successfully transferred into chokecherry. The transformation will be further confirmed by polymerase chain reaction (PCR) and Southern blot analyses.

scholarly journals (280) Plant Regeneration of Periwinkle (Catharanthus roseus) from In Vitro Leaf Tissues

HortScience ◽  
2005 ◽  
Vol 40 (4) ◽  
pp. 1050E-1051
Author(s):  
Wenhao Dai ◽  
Victoria Jacques
Keyword(s):  
Disease Transmission ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Shoot Formation ◽  
Lymphocytic Leukemia ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Bedding Plant ◽  
Leaf Tissues

Periwinkle, a perennial commonly used as a summer bedding plant, is known as the source of vinca alkaloids used to treat lymphocytic leukemia and Hodgkin's disease. It is also one of the natural hosts of many phytoplasma diseases, such as X-disease of major Prunus species, aster yellows, and ash yellows diseases. Therefore, periwinkle is an ideal plant species for phytoplasma disease research, such as disease transmission, species resistance, and resistant gene screening. Periwinkle tissue culture was established by incubating sterile seeds in hormone-free Murashige and Skoog (MS) medium. Plants were successfully regenerated from in vitro leaf tissues of periwinkle. Adventitious shoots were induced when leaf tissues were cultured on Murashige and Skoog (MS) medium or woody plant medium (WPM) supplemented with benzyladenine (BA) and naphthaleneacetic acid (NAA). Nearly 75% of leaf explants produced shoots in both media with 10–20 μm BA and 1 μm NAA. A mean of 4.3 shoots was produced from each explant cultured on WPM, whereas only 2 shoots were produced on MS medium under 16-h photoperiod. Leaf explants under dark treatment for 2 weeks produced big callus only, indicating that light is necessary for shoot formation. Most adventitious shoots were induced from the joint of leaf blade and petiole. In vitro shoots (>1.5 cm) were easily rooted in half-strength MS medium. Addition of NAA dramatically increased root number, with more than 20 roots being induced in 5 μm NAA medium. Rooted plants were transferred to potting medium and grown in a greenhouse.

scholarly journals Influence of different pathways of regeneration on genetic and phytochemical instability of Curculigo orchioides

2015 ◽  
Vol 24 (2) ◽  
pp. 173-189
Author(s):  
Parameswari Alagar ◽  
Rahul Raveendran Nair ◽  
Ganesh Doss
Keyword(s):  
Low Frequency ◽  
Leaf Explants ◽  
Direct Regeneration ◽  
Terminal Part ◽  
Curculigo Orchioides ◽  
Leaf Tissues ◽  
Micropropagated Plants ◽  
Medicinal Properties ◽  
Explant Analysis

Micropropagated plants of Curculigo orchioides Gaertn. through five different modes of regeneration were evaluated for their genetic stability and major flavanoids content that contribute to medicinal properties of the rhizome. Leaf explants that produced callus and bulbils have produced plantlets at a higher frequency. However, direct regeneration of plants from leaf tissues without intervening callus produced a low frequency of plantlets. Explants of rhizome produced healthy plantlets directly from the terminal part of the explant. Plantlets regenerated through five different modes of regeneration did not show any morphological variation. The duration of regeneration was varying from 60 to 145 days depending upon the type of the explant. Analysis of genomic DNA among the plantlets regenerated through five different modes of micropropagation revealed about 66% polymorphism. Major flavanoids such as rutin, ferulic acid, caffeic acid, quercitrin that contributed to medicinal properties of the rhizomes differed quantitatively among the plantlets regenerated through different pathways. Present study suggests that plantlets regenerated directly from the rhizome as well as from the leaf tips of in vitro grown plants are stable genetically and phytochemically.Plant Tissue Cult. & Biotech. 24(2): 173-189, 2014 (December)

scholarly journals Micropropagation of Betula platyphylla ‘Fargo’ via Shoot Tip Culture and Regeneration from Leaf Tissues

2000 ◽  
Vol 18 (2) ◽  
pp. 119-122 ◽  
Author(s):  
Zong-Ming Cheng ◽  
Jeffrey P. Schnurr ◽  
Wenhao Dai
Keyword(s):  
Woody Plant ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Betula Platyphylla ◽  
Ex Vitro ◽  
White Birch ◽  
Dark Treatment ◽  
Woody Plant Medium ◽  
Leaf Tissues

Abstract A micropropagation system was developed for mass propagation of ‘Fargo’® (Dakota Pinnacle™), a newly released cultivar of Asian white birch (Betula platyphylla). Shoot tips from the mature, 7-year-old tree were established on 75% strength Murashige and Skoog medium supplemented with 0.1 μM thidiazuron. The greatest shoot proliferation occurred on Woody Plant Medium supplemented with 2.2 μM benzyladenine (BA), solidified with 6.5 g/liter agar, and cultured at 24C (75F). Microshoots were rooted in vitro or ex vitro followed by establishment in the greenhouse. A system to regenerate plantlets from leaves of aseptically cultured shoots was also developed. The optimum conditions for shoot regeneration from leaves included a 2-week dark treatment before exposure to a 16/8 hour light/dark photoperiod, use of large and healthy leaf explants, and culture on the Woody Plant Medium containing 20.0 or 30.0 μM BA. The regenerated shoots proliferated on the micropropagation medium and were divided, and the resulting shoots were rooted ex vitro and acclimated in greenhouse conditions.

The role of silicon in forages: A quantitative X-Ray study

1987 ◽  
Vol 45 ◽  
pp. 416-417
Author(s):  
Janet H. Woodward ◽  
D. E. Akin
Keyword(s):  
Sodium Acetate ◽  
Dry Matter ◽  
Acetate Buffer ◽  
Plant Tissues ◽  
Sodium Acetate Buffer ◽  
X Ray ◽  
Dry Matter Digestibility ◽  
Percent Composition

Silicon (Si) is distributed throughout plant tissues, but its role in forages has not been clarified. Although Si has been suggested as an antiquality factor which limits the digestibility of structural carbohydrates, other research indicates that its presence in plants does not affect digestibility. We employed x-ray microanalysis to evaluate Si as an antiquality factor at specific sites of two cultivars of bermuda grass (Cynodon dactvlon (L.) Pers.). “Coastal” and “Tifton-78” were chosen for this study because previous work in our lab has shown that, although these two grasses are similar ultrastructurally, they differ in in vitro dry matter digestibility and in percent composition of Si.Two millimeter leaf sections of Tifton-7 8 (Tift-7 8) and Coastal (CBG) were incubated for 72 hr in 2.5% (w/v) cellulase in 0.05 M sodium acetate buffer, pH 5.0. For controls, sections were incubated in the sodium acetate buffer or were not treated.

FACTORS AFFECTING THE REGENERATION OF PEPPER (CAPSICUM ANNUUM L.)

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1120G-1120
Author(s):  
J. L. Jacobs ◽  
C. T. Stephens
Keyword(s):  
Growth Hormone ◽  
Silver Nitrate ◽  
Callus Formation ◽  
Leaf Explants ◽  
Regeneration System ◽  
Factors Affecting ◽  
Nitrate Concentrations ◽  
Leaf Differentiation ◽  
Whole Plants

Several growth hormone combinations and silver nitrate concentrations were examined for their effect on regeneration of different pepper genotypes. Primary leaf explants from in vitro seedlings were cultured on a revised Murashige and Skoog medium supplemented with auxin, cytokinin and 1.6% glucose. Combinations of different concentrations of indole-3-acetic acid (IAA), 0-5 mg/l, and 6-benzylaminopurine (BAP), 0-5 mg/l, were tested to determine the most effective medium for shoot primordium formation. Experiments with IAA and BAP did not result in a specific growth hormone combination appropriate for regeneration of all genotypes tested. Of the silver nitrate concentrations tested, 10 mg/l resulted in the best shoot and leaf differentiation and reduced callus formation. Differences in organogenic response of individual genotypes were evaluated on a single regeneration medium. Whole plants were regenerated from 11 of 63 genotypes examined. Based on these experiments, a reproducible regeneration system for pepper was developed with a total of 500 plants regenerated to date.

Development of an Agrobacterium-mediated Transformation System for `Beurre Bosc' Pear

HortScience ◽  
1998 ◽  
Vol 33 (3) ◽  
pp. 461d-461
Author(s):  
Richard L. Bell ◽  
Ralph Scorza ◽  
Chinnathambi Srinivasan
Keyword(s):  
Shoot Proliferation ◽  
Induction Medium ◽  
Shoot Induction ◽  
Transformation System ◽  
Gus Histochemical Assay ◽  
Young Leaves ◽  
Leaf Tissues ◽  
Shoot Induction Medium ◽  
Basal Salts

An efficient regeneration/transformation system was developed for `Beurre Bosc' pear. Young leaves were harvested from in vitro shoots proliferated on a medium containing MS basal salts and 5 BAP, 0.5 μM IBA, and 0.6M3. Shoot regeneration was optimized using a modification of the medium of Chevreau and Leblay (1993). Explants were cultured on shoot induction medium contained 10 μM TDZ and 1 μM IBA for 4 weeks in the dark, and then transfered to a similar, but auxinless, regeneration medium until shoots developed, usually after an additional 4 to 8 weeks. Leaf tissues were transformed by co-cultivation for 3 days with Agrobacterium tumefaciens EHA101 carrying a pGA482 plasmid containing NPTII, GUS, and rolC genes, followed by cultivation on SIM containing 300 mg/L timentin. Putative transgenic plants were selected on shoot induction medium containing 80mg/L kanamycin, and multiplied on shoot proliferation medium. Four clones were confirmed as transgenic using the GUS histochemical assay and Southern blots for the NPTII and rolC genes. Plants of each clone have been rooted and successfully transfered to the greenhouse for further analysis of gene expression.

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