scholarly journals Expression pattern of in vitro organogenesis-associated genes as transcriptional marker in Indian Sandalwood (Santalum album L.).

2021 ◽  
Author(s):  
Gareema Pandey ◽  
Arpan Modi ◽  
Shikha Shah ◽  
Ghanshyam Patil ◽  
Subhash Narayanan
Keyword(s):  
Expression Pattern ◽  
Callus Formation ◽  
Expression Patterns ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Santalum Album ◽  
Mass Propagation ◽  
Indirect Pathway ◽  
Indirect Organogenesis

Abstract Indian sandalwood (Santalum album L.) is an expensive wood that requires reproducible method for mass propagation to ensure consistent production and sustainable use of sandalwood. For mass propagation of sandalwood, plant organogenesis requires different combinations of the tissue culture medium. The media is composed of exogenous phytohormones which decides the explant's morphological stages such as shooting or rooting induction. Early prediction of morphological stage from explant can potentially help in selecting the exogenous phytohormones combinations thereby saving time and resources for mass sandalwood propogation. An efficient protocol for the direct and indirect organogenesis (up to shooting development phase) of sandalwood were developed using Woody Plant Media (WPM). WPM supplemented with various concentrations of 6-Bezylaminopurine (BAP) and 1-Naphthaleneacetic acid (NAA) were tested for direct organogenesis, while different treatments consisting of various levels of 2,4-dichlorophenoxyacetic acid (2,4-D), NAA, BAP, Adenine sulphate (ADS), glycine and potassium nitrate were tested for indirect organogenesis. Three stages of leaf development were selected viz., the leaf just after inoculation in WPM media, initial stage of callus formation from leaf and shoot formation for expression pattern analysis. The targeted genes were Alternative oxidase (ao), Late embryogenesis abundant (lea), Cytochrome P450 (cyt-p450), ABC transporter (abct), and Serine-threonine phosphatase (stp) which are associated with in vitro organogenesis. The expression patterns were evaluated to identify a transcription marker. During the initial stages of organogenesis, ao, cyt-p450 and abct showed no/little change in expression in the direct pathway but up-regulation of ao and abct and downregulation of cyt-p450 were observed in the indirect pathway. Expression of lea was increased up to 70-fold during direct and dropped to half during indirect organogenesis.

scholarly journals In vitro morphogenesis of Syngonanthus mucugensis Giul: subsp. mucugensis

2011 ◽  
Vol 35 (3) ◽  
pp. 502-510 ◽  
Author(s):  
Alone Lima-Brito ◽  
Sheila Vitória Resende ◽  
Carolina Oliveira de Cerqueira Lima ◽  
Bruno Matos Alvim ◽  
Claudia Elena Carneiro ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
Callus Formation ◽  
Economic Value ◽  
Direct Organogenesis ◽  
Naphthalene Acetic Acid ◽  
Herbaceous Plant ◽  
Indirect Organogenesis ◽  
In Vitro Morphogenesis ◽  
Different Types

Syngonanthus mucugensis Giul. subsp. mucugensis is an herbaceous plant with significant economic value in the ornamental dry flower business. The restricted occurrence of the municipality Mucugê-BA, Brazil, exclusively associated with extractive exploitation, has considered this species as endangered. The objective of this work was to evaluate the organogenic potential of three different types of S. mucugensis subsp. mucugensis explants to promote the development of an alternative method to the propagation of the genetic resources of this important plant. The morphogenetic capacities of the leaf, stem and root this species was tested using Murashige and Skoog culture medium at half salt concentration and different concentrations of growth of regulators benzylaminopurine - BAP (0.00; 2.22 and 4.44 µM), and naphthalene acetic acid - NAA (0.00; 1.34 and 2.68 µM). The morphoanatomic events that lead to formation of shoots were described. Stems proved to be the best source of explants, showing 58.75% regeneration of shoot by direct organogenesis in the absence of growth regulators, and 32.18 and 47.55% of shoot regeneration by indirect organogenesis in the presence of 2.22 and 4.44 µM BAP, respectively. As for leaves, there was callus formation, but without regenerating shoots. Morphogenesis was not observed when roots were used as explants. The histological analyses showed that shoot regeneration in S. mucugensis subsp. mucugensis occurred both indirectly, by unorganized tissue differentiation, and directly through returning to merismatic activity in differentiated mature cells and preexisting bud proliferation.

scholarly journals Massive in vitro Cloning of Sandalwood (Santalum album Linn.) via Cultured Nodal Segments

2019 ◽  
pp. 1-14
Author(s):  
D. Bele ◽  
Nishi Mishra ◽  
Sushma Tiwari ◽  
M. K. Tripathi ◽  
G. Tiwari
Keyword(s):  
Somatic Embryogenesis ◽  
Plantlet Regeneration ◽  
Nodal Segments ◽  
Direct Somatic Embryogenesis ◽  
Direct Organogenesis ◽  
Santalum Album ◽  
Ms Medium ◽  
Regeneration Medium ◽  
Indirect Organogenesis

Nodal segments of sandalwood were cultured on MS medium amended with different plant growth regulators in varying concentrations to search out higher in vitro response leading to plantlet regeneration via somatic embryogenesis and/or organogenesis. Higher proportion of direct somatic embryogenesis, number(s) of somatic embryo per explant and plantlet regeneration via direct organogenesis were recorded on MS medium supplemented with a moderate concentration of TDZ (1.0 mgl-1) in combination with comparatively a lower concentration of NAA (0.5 mgl-1). A relative higher concentration of BAP (1.0-2.0 mgl-1) in combination with a lower concentration of NAA (0.5 mgl-1) promoted frequency of indirect somatic embryogenesis. Ratio of organ formation directly from surface of cultured explants was recovered from culture medium fortified with a higher concentration of BA at the concentration of 4.0 mgl-1 in combination with a lower concentration of NAA (0.5 mgl-1). Maximum plantlets regenerated via somatic embryogenesis (direct and/or indirect) on regeneration medium supplemented with 2.0 mgl-1TDZ  in combination with 1.0 mg l-1GA3, while plantlets in higher frequencies via indirect organogenesis was attained with regeneration medium amended with comparatively lower concentration of TDZ (1.0 mg l-1) in combination with 0.5 mgl-1 GA3 and 0.5     mgl-1 NAA. The plantlets were transferred to pots and hardened in Environmental Growth Cabinet and Net House during preliminary weaning period and transferred to field successfully. Morphologically normal plants were recovered.

scholarly journals Direct Shoot Regeneration from Vaccinium pahalae (Ohelo) and V. myrtillus (Bilberry) Leaf Explants

HortScience ◽  
1996 ◽  
Vol 31 (7) ◽  
pp. 1225-1228 ◽  
Author(s):  
Rida A. Shibli ◽  
M.A.L. Smith
Keyword(s):  
Callus Formation ◽  
Leaf Explants ◽  
Shoot Proliferation ◽  
Direct Organogenesis ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
Direct Shoot Regeneration ◽  
The Media ◽  
2,4 Dichlorophenoxyacetic Acid

Ohelo (V. pahalae Skottsb.) and bilberry (V. myrtillus L.) shoots were regenerated via direct organogenesis from whole leaves and leaf sections and also from hypocotyl explants of bilberry. Explants preincubated for 1 to 2 weeks in darkness yielded ≈75% regeneration frequencies and the highest number of regenerating shoots/explant on TDZ-supplemented media (0.9 to 2.7 μm). When 2iP or zeatin were substituted as the cytokinin source, frequencies of regeneration and shoot productivity were significantly lower. Explants held under constant illumination (no dark pretreatment) had significantly lower regeneration frequencies in all tested cytokinin-supplemented media. 2,4-D stimulated callus formation, but did not support regeneration from vegetative explants. Cells from callus and suspension cultures did not exhibit regeneration in any of the media that supported organogenesis from leaves. Regenerants were successfully micropropagated, although callus formation caused by zeatin and high 2iP levels interfered with shoot proliferation. Zeatin induced hyperhydricity in shoots from both species, but more severely in ohelo. Ex vitro rooting after treatment with 4.9 μm IBA or 5.4 μm NAA was 95% and 60% successful for bilberry and ohelo, respectively, and plants were readily acclimatized after an interval in a fog chamber. Bilberry microshoots also rooted in vitro in the absence of growth regulator treatment. Chemical names used: 1H-indole-3-butanoic acid (IBA); N-(3-methyl-2-butenyl)-1-H-purine-6-amine (2iP); 6-furfurylaminopurine (kinetin); 1-naphthaleneacetic acid (NAA); thidiazuron=1-phenyl-3-(1,2,3-thiadiazio-5-yl)urea (TDZ); 2,4-dichlorophenoxyacetic acid (2,4-D); 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

scholarly journals In vitro morphogenesis of Physalis ixocarpa Brot ex. Horm

2021 ◽  
Vol 51 ◽  
Author(s):  
Domitzel Zagal Alvarado ◽  
Andressa Priscila Piancó Santos Lima ◽  
José Raniere Ferreira de Santana ◽  
Alone Lima-Brito
Keyword(s):  
Tissue Culture ◽  
Experimental Design ◽  
Cotyledonary Node ◽  
Direct Organogenesis ◽  
Naphthaleneacetic Acid ◽  
Indirect Organogenesis ◽  
In Vitro Morphogenesis ◽  
Important Species ◽  
Randomized Experimental Design

ABSTRACT Physalis ixocarpa Brot. ex Horm. is considered the most economically important species of the genus. Tissue culture is pointed out as a strategy for its propagation, but researches indicate that in vitro responses are genotype-dependent. This study aimed to evaluate the in vitro morphogenesis of the P. ixocarpa green and purple varieties, in view of the massive propagation of the species. The morphogenic capacity of the explants cotyledonary node, hypocotyl and cotyledon was evaluated in Murashige & Skoog medium supplemented with benzylaminopurine - BAP (0.00, 2.5, 5.0, 7.5 or 10.0 μM) and naphthaleneacetic acid - NAA (0.00 or 2.5 μM), using a completely randomized experimental design, in a 3 x 5 x 2 factorial scheme, with 30 treatments for each variety. The number of shoots per direct and indirect organogenesis and the percentage of explants with callus were analyzed. The in vitro morphogenetic expression of P. ixocarpa is influenced by the type of explant and by the plant regulators BAP and NAA. The cotyledonary node explant is efficient for the production of shoots via direct organogenesis in the two varieties studied.

Selectivity of auxin for induction and growth of callus from excised embryo of spring and winter wheat

1984 ◽  
Vol 62 (7) ◽  
pp. 1393-1397 ◽  
Author(s):  
M. D. Zhou ◽  
T. T. Lee
Keyword(s):  
Winter Wheat ◽  
Chinese Spring ◽  
Shoot Growth ◽  
Callus Formation ◽  
Triticum Aestivum L ◽  
Dichlorophenoxyacetic Acid ◽  
Naphthaleneacetic Acid ◽  
2,4 Dichlorophenoxyacetic Acid ◽  
Methoxybenzoic Acid

The callus-promoting activity of most commonly known as well as some rarely tested auxins was compared with that of 2,4-dichlorophenoxyacetic acid (2,4-D) for in vitro culture of the excised embryo of spring and winter wheat (Triticum aestivum L.), cv. Chinese Spring and cv. Fredrick. Different auxins in a concentration range from 1 to 50 μM showed widely different activities. Also the two wheat cultivars responded differently to the auxins. When rapid callus formation with limited root growth was used as the basis for comparison, 2-(2-methyl-4-chlorophenoxy)propionic acid (2-MCPP), α-naphthaleneacetic acid, 3,6-dichloro-2-methoxybenzoic acid (dicamba), 4-amino-3,5,6,trichloropicolinic acid (picloram), γ-(2,4-dichlorophenoxy)butyric acid, 2,4,5-trichlorophenoxyacetic acid, and 2,4,5-trichlorophenoxypropionic acid, in the order of effectiveness, were superior to 2,4,-D for callus induction from the embryo of 'Chinese Spring,' although the concentration required was higher than that of 2,4-D. For the winter wheat 'Fredrick,' however, only picloram, dicamba, and 2-MCPP performed as well as 2,4-D. All auxins tested promoted shoot growth; 2-methyl-4-chlorophenoxypropionic acid was most effective for 'Chinese Spring,' whereas picloram was most effective for 'Fredrick.'

scholarly journals Factors Affecting the Regeneration, via Organogenesis, and the Selection of Transgenic Calli in the Peach Rootstock Hansen 536 (Prunus persica × Prunus amygdalus) to Express an RNAi Construct against PPV Virus

Plants ◽  
2019 ◽  
Vol 8 (6) ◽  
pp. 178 ◽  
Author(s):  
Sabbadini ◽  
Ricci ◽  
Limera ◽  
Baldoni ◽  
Capriotti ◽  
...  
Keyword(s):  
Shoot Regeneration ◽  
Prunus Persica ◽  
Fluorescent Protein ◽  
In Vitro Regeneration ◽  
Naphthaleneacetic Acid ◽  
Prunus Amygdalus ◽  
Indirect Organogenesis ◽  
Egfp Gene ◽  
Rnai Construct

Prunus spp. is one of the most recalcitrant fruit tree species in terms of in vitro regeneration and transformation, mostly when mature tissues are used as explants. The present study describes the in vitro regeneration via indirect organogenesis, and Agrobacterium tumefaciens-mediated transformation of the peach rootstock Hansen 536 (Prunus persica × Prunus amygdalus) through the use of meristematic bulks (MBs) as starting explants. Efficient adventitious shoot regeneration was obtained when Hansen 536 MBs were cultured on an optimized medium consisting of modified McCown Woody Plant medium (WPM) enriched with 4.4 M 6-Benzyladenine (BA), 0.1 M 1-Naphthaleneacetic acid (NAA) and 6.0 g L−1 plant agar S1000 (B&V). MB slices were used later as starting explants for Agrobacterium-mediated transformation to introduce an RNAi construct “ihp35S-PPV194” against PPV virus. Transgenic events were identified by both green fluorescent protein (GFP) screening and kanamycin selection at different concentrations (0, 17 or 42 M). GFP-fluorescent proliferating callus lines were selected and confirmed to stably express the ihp35S-PPV194::eGFP gene construct by molecular analysis. Although shoot regeneration from these transgenic calli has not been obtained yet, this represents one of the few examples of successful attempts in peach genetic transformation from somatic tissues, and also serves as a useful in vitro system for future gene functional analysis in peach.

Mesenchymal Stem Cell Differentiation Markers Shared with Soft Tissue Sarcomas

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 4755-4755
Author(s):  
Stefan Wirths ◽  
Hans-Joerg Buehring ◽  
Lothar Kanz ◽  
Joerg T Hartmann ◽  
Hans-Georg Kopp
Keyword(s):  
Stem Cells ◽  
Soft Tissue ◽  
Expression Pattern ◽  
Cell Lines ◽  
Expression Profiles ◽  
Expression Patterns ◽  
Soft Tissue Sarcomas ◽  
Stem Cell Differentiation ◽  
Antibody Panel

Abstract Malignant tumors are hypothesized to harbor small populations of self-renewing cancer stem cells. Targeting these cells may be the decisive step to overcome treatment resistance and achieve tumor eradication in cancer patients. Advanced soft tissue sarcomas (STS) are rare tumors with a dismal prognosis and a small number of systemic treatment options. STS may originate from mesenchymal stem cells (MSC); the latter have mainly been isolated from adult bone marrow (BM) as non-hematopoietic, self-renewing cells whose in vitro progeny comprises osteoblasts, chondroblasts, myocytes, and adipocytes. While in vitro expression profiles of MSC have been investigated extensively, the in vivo counterparts of MSC are still hypothetical. To target rare human cell BM populations including MSC, an exclusive antibody panel was developed. The target antigens include platelet-derived growth factor receptor-β (CD140b), HER-2/erbB2 (CD340), the recently described W8B2 antigen as well as several surface antigens identified by novel antibodies. To define the expression pattern of MSC-markers in STS, three STS cell lines were tested for expression of these antigens. In addition, snap-frozen primary STS sections were analyzed by immunohistochemistry using the same antibody panel. All cell lines revealed expression of selected markers including CD340, W8B2, and CD140b. Several MSC markers were restricted to a subpopulation of cells. In addition, leiomyosarcoma cells displayed a different expression pattern as compared to liposarcoma and Ewing’s sarcoma cells. Results of immunohistochemistry analysis of primary leiomyosarcoma tumor samples correlated strongly with expression patterns established by FACS analysis. However, important cytoarchitectural features regarding selected markers were revealed by immunohistochemistry: while primary leiomyosarcomas displayed uniform expression of W7C6, HEK3D6, CD10, and CD318, other markers such as CD34, W5C5, and 57D2 were expressed by tumor endothelia only. Moreover, a population of perivascular tumor cells was found to express the MSC-markers W4A5, W8B2, CD140b, W3D5, and W5C4. Novel MSC-markers are expressed by subpopulations in STS cell lines as well as in primary sarcoma tissue. Further studies on the functional significance of these phenotypical studies are underway and may help to identify novel specific targets recognizing the self-renewing STS-compartment.

scholarly journals Micropropagation of Clerodendrum phlomidis L.F.

2016 ◽  
Vol 24 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Mafatlal M. Kher ◽  
Deepak Soner ◽  
Neha Srivastava ◽  
Murugan Nataraj ◽  
Jaime A. Teixeira da Silva
Keyword(s):  
Callus Formation ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Nodal Explants ◽  
Axillary Shoot ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Axillary Shoot Proliferation ◽  
Clerodendrum Phlomidis

Abstract Clerodendrum phlomidis L. f. is an important medicinal plant of the Lamiaceae family, particularly its roots, which are used for various therapeutic purposes in a pulverized form. The objective of this study was to develop a standard protocol for axillary shoot proliferation and rooting of C. phlomidis for its propagation and conservation. Nodal explants were inoculated on Murashige and Skoog (MS) medium that was supplemented with one of six cytokinins: 6-benzyladenine, kinetin, thidiazuron, N6-(2-isopentenyl) adenine (2iP), trans-zeatin (Zea) and meta-topolin. Callus induction, which was prolific at all concentrations, formed at the base of nodal explants and hindered shoot multiplication and elongation. To avoid or reduce callus formation with the objective of increasing shoot formation, the same six cytokinins were combined with 4 μM 2,3,5-tri-iodobenzoic acid (TIBA) alone or in combination with 270 μM adenine sulphate (AdS). Nodal explants that were cultured on the medium supplemented with 9.12 μM Zea, 4 μM TIBA and 270 μM AdS produced significantly more and longer shoots than on medium without TIBA and AdS. Half-strength MS medium supplemented with 8.05 μM α-naphthaleneacetic acid was the best medium for root formation. Most (75%) in vitro rooted plantlets were successfully acclimatized under natural conditions.

scholarly journals In-vitro Regeneration from Direct and Indirect Organogenesis of Crescentia alata Kunth an Important Multipurpose Medicinal Tree

2021 ◽  
pp. 48-58
Author(s):  
R. Abinaya
Keyword(s):  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Basal Medium ◽  
Direct Organogenesis ◽  
Ms Medium ◽  
Indirect Organogenesis ◽  
Medicinal Tree ◽  
Short Period ◽  
In Vitro Clonal Propagation

In this present work, an in-vitro regeneration protocol for Crescentia alata (C. alata) was developed using various explants on Murashige and Skoog (MS) medium augmented with different concentrations and combinations of plant growth regulators (PGRs) for direct and indirect regeneration. The direct organogenesis was established from nodes and internodes on MS medium supplemented with cytokinins and auxins. The indirect organogenesis via callus phase was obtained from leaf, nodes and internodes on MS medium supplemented with different concentrations of PGRs. The high frequency shoot organogenesis were achieved directly from nodal explants were cultured on MS medium supplemented with 3.0 mg/L BAP+0.5 mg/L KIN +1.0 mg/L NAA. Indirect organogenesis callogenic frequency was optimized at the concentration of MS medium containing 1.0 mg/L BAP + 5.0 mg/L IAA. The callus was obtained from all the explants were used, among these explants internodal explants gave best result on MS medium supplemented with different concentrations of cytokinins and auxins for indirect organogenesis experiment. Indirect organogenesis the highest number of shoot regeneration was obtained in MS Basal Medium with 4.0 mg/L BAP + 0.5 mg/L KIN + 2.0 mg/L NAA from internodal explants. For root formation the regenerative shoots which were sub cultured on MS medium containing different ratios of auxins. The rooted plantlets were transferred successfully to the pots containing sterilized soil and were successfully hardened at greenhouse condition for 20 days then exposed to the natural environment. This is the first successful micropropagation report of an efficient and rapid in-vitro clonal propagation protocol for C. alata by direct and indirect shoot organogenesis through various explants, which can be employed for conservation of this important medicinal tree species as well as the utilization of an biologically important active biomolecules. This protocol can be very useful to obtain plants from various explants, without the requirement of meristematic regions, enabling the obtainment of a higher number of plants in short period.

scholarly journals Growth and Photosynthetic Capability of Momordica grosvenori Plantlets Grown Photoautotrophically in Response to Light Intensity

HortScience ◽  
2009 ◽  
Vol 44 (3) ◽  
pp. 757-763 ◽  
Author(s):  
Meijun Zhang ◽  
Duanduan Zhao ◽  
Zengqiang Ma ◽  
Xuedong Li ◽  
Yulan Xiao
Keyword(s):  
Callus Formation ◽  
Dry Weight ◽  
Control Treatment ◽  
Photon Flux ◽  
Photon Flux Density ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Light Intensities ◽  
Four Levels

Momordica grosvenori plantlets were cultured in vitro for 26 d on sucrose- and hormone-free Murashige and Skoog (MS) medium with four levels of photosynthetic photon flux density (PPFD), namely 25, 50, 100, or 200 μmol·m−2·s−1, and a CO2 concentration of 1000 μmol·mol−1 in the culture room [i.e., photoautotrophic micropropagation (PA) treatments]. The control treatment was a photomixotrophic culture using MS medium containing sucrose and NAA with a CO2 concentration of 400 μmol·mol−1 in the culture room and a PPFD of 25 μmol·m−2·s−1. Based on the results, a second experiment was conducted to investigate the effects of α-naphthaleneacetic acid (NAA) and sucrose on callus formation. For this, plantlets were grown in the absence and presence of either NAA or sucrose. Compared with the control, the PA plantlet had a well-developed rooting system, better shoot, greater chlorophyll content, and higher electron transport rate and the ex vitro survival percentage was increased by 31%. Both sucrose and NAA stimulated callus formation on the shoot bases of control plantlets, whereas calluses did not form on the plantlets grown in sucrose- and hormone-free medium. The stronger light intensities increased the fresh and dry weight of plantlets. A PPFD of 100 μmol·m−2·s−1 was more suitable for the growth of M. grosvenori plantlets. Therefore, photoautotrophic plantlets grown at high light intensities would be better suited to the intense irradiance found in sunlight.

Sign in / Sign up

Export Citation Format

Share Document