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 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 Agrobacterium-Mediated Transformation of Chokecherry (Prunus virginiana L.)

HortScience ◽  
2007 ◽  
Vol 42 (1) ◽  
pp. 140-142 ◽  
Author(s):  
Wenhao Dai ◽  
Victoria Magnusson ◽  
Chris Johnson
Keyword(s):  
Acetic Acid ◽  
Southern Blot ◽  
Blot Analysis ◽  
Southern Blot Analysis ◽  
Infected Leaf ◽  
Naphthalene Acetic Acid ◽  
Prunus Virginiana ◽  
Leaf Tissues ◽  
Half Strength

Chokecherry (Prunus virginiana L.) was transformed using Agrobacterium tumefaciens strain EHA105 harboring binary vector pBI121 carrying the neomycin phosphotransferase gene (nptII) and β-glucuronidase (GUS) gene (uidA). Plants were regenerated from the Agrobacterium-infected leaf tissues through organogenesis on woody plant medium (WPM) supplemented with MS (Murashige and Skoog) vitamins, 10 μm 6-benzyladenine (BA), and 250 mg·L−1 cefotaxime plus 500 mg·L−1 carbenicillin plus 15 mg·L−1kanamycin (CCK15). Transformation was verified with polymerase chain reaction (PCR) and Southern blot analysis. Four of 150 (2.67%) initial explants produced GUS- and PCR-positive shoots. Southern blot analysis confirmed that the transgenes were integrated into the chokecherry genome. Transgenic in vitro shoots were rooted in half-strength MS medium containing 10 μm naphthalene acetic acid. Rooted plants were transferred to potting mix and grown in the greenhouse. This research shows a potential for future improvement of chokecherry and other Prunus species. Chemical names used: 6-benzyladenine (BA), naphthalene acetic acid (NAA), acetosyringone (AS), 5-Bromo-4-chloro-3-indoxyl-beta-D-glucuronide cyclohexylammonium (X-Glu), cefotaxime, carbenicillin, kanamycin.

scholarly journals Establishment of a Plant Regeneration and Genetic Transformation System of Panax ginseng C.A. Meyer

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 572d-572
Author(s):  
Hak-Tae Lim ◽  
Haeng-Soon Lee ◽  
Tage Eriksson
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Genetic Transformation ◽  
Adventitious Shoots ◽  
Shoot Multiplication ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Genetic Transformation System ◽  
Regeneration Systems

Plant regeneration of ginseng has been known to be difficult, and there are a few reports on plant regeneration of ginseng via somatic embryogenesis. In vitro flowering has, however, been one of the major drawbacks in these regeneration systems in which BA and GA3 were included in germination and shoot multiplication media. Multiplication of adventitious shoots from a single somatic embryo, abnormal morphology, and vitrified shoots were also observed. All these facts have made successful acclimatization of ginseng plantlets difficult. The purposes of this study were 1) to establish the plant regeneration system via organogenesis, 2) to improve normal plant regeneration via somatic embryogenesis, 3) to improve the efficiency of plant regeneration from protoplast culture, 4) to understand the acclimatization process, 5) to develop effective genetic transformation protocol. Data in relation with all these studies are presented in detail.

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 An Efficient Tissue Culture Regeneration System for Georgia Plume, Elliottia racemosa, a Threatened Georgia Endemic

HortScience ◽  
2008 ◽  
Vol 43 (2) ◽  
pp. 447-453 ◽  
Author(s):  
Seong Min Woo ◽  
Hazel Y. Wetzstein
Keyword(s):  
Tissue Culture ◽  
Plant Regeneration ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Dry Weight ◽  
Medium Composition ◽  
Induction Medium ◽  
Ex Vitro ◽  
Regeneration System

Georgia plume, Elliottia racemosa Muhlenb. ex. Elliott, is an extremely rare small tree or shrub endemic to Georgia, which is being severely affected by habitat loss and lack of sexual recruitment. In vitro plant regeneration of Georgia plume has not been previously reported and may be a method for the conservation and propagation of this threatened species. Studies evaluated the effects of sterilization methods, explant types, medium composition, and light environment on plant regeneration. An efficient plant regeneration system was developed in which adventitious shoot buds were induced using young, expanding leaf explants placed on an induction medium supplemented with 10 μm thidiazuron and 5 μm indole-3-acetic acid with Gamborg's B5 salts. Shoot elongation was promoted on a medium with 25 μm (2-isopentenyl) adenine incorporated into Woody Plant Medium. In vitro rooting studies evaluated continuous and pulse auxin treatments and ex vitro rooting trials after KIBA (indole-3-butric acid, potassium salt) dips. A 5-day pulse treatment on 100 or 150 μm indole-3-butyric acid produced ≈90% rooting of shoots with greater shoot and root dry weight than other pulse times. High rooting frequencies were obtained under in vitro and ex vitro conditions with over 85% survival of plantlets transferred to greenhouse conditions. The culture protocol was found to be effective with material collected from mature specimens in the wild from divergent populations. Tissue culture appears to be a promising approach for the propagation and conservation of this rare and threatened plant.

scholarly journals Plant Regeneration of Periwinkle (Catharanthus roseus) via Organogenesis

HortScience ◽  
2008 ◽  
Vol 43 (3) ◽  
pp. 832-836 ◽  
Author(s):  
Andrea Swanberg ◽  
Wenhao Dai
Keyword(s):  
Plant Regeneration ◽  
Shoot Regeneration ◽  
Woody Plant ◽  
Naphthalene Acetic Acid ◽  
Regeneration System ◽  
Dark Treatment ◽  
Regeneration Rate ◽  
Shoot Production ◽  
Leaf Tissues

Two periwinkle cultivars, Pacific Coral (P1) and Sunstorm Rose (P2), were used for development of a plant regeneration system. Leaf and internodal explants collected from in vitro plants were plated onto woody plant medium (WPM) using a factorial arrangement of 6-benzyladeine (BA) and 1-naphthalene acetic acid (NAA). Shoots were successfully regenerated. Shoot production from leaf tissues was minimal for all cultivars, whereas internodal tissues showed variable rates of regeneration depending on the hormone combination. Cultivar P1 showed the maximum regeneration rate (73.3%) when internodal explants, 4 to 6 mm in length, were placed on WPM containing 5 μm BA and 5 μm NAA. Cultivar P2 showed a regeneration rate of 56.7% with a combination of 20 μm BA and 10 μm NAA. Shoot regeneration rate increased as the internodal explant size increased for P2; however, the regeneration rate decreased when the explant size was greater than 7 mm for P1. The shoot regeneration rate decreased as the period of the dark treatment of internodal explants increased in both P1 and P2. The antibiotics carbenicillin (Carb) and cefotaxime (Cef) had little effect on shoot regeneration. There was a slightly higher rate observed for P1 when Cef was added into the medium, whereas P2 showed a decrease with the addition of Cef. Carb showed no significant effect on shoot regeneration for both cultivars. Addition of both Carb and Cef to the medium slightly inhibited shoot regeneration.

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.

scholarly journals Seed Germination and in vitro Plant Regeneration through Callus Culture of Two Lychnis Species

2015 ◽  
Vol 25 (1) ◽  
pp. 1-12 ◽  
Author(s):  
Kee Hwa Bae ◽  
Eui Soo Yoon
Keyword(s):  
Plant Regeneration ◽  
Seed Germination ◽  
Callus Induction ◽  
In Vitro Propagation ◽  
Ornamental Plants ◽  
Leaf Explants ◽  
Adventitious Shoot ◽  
Temperature Treatment ◽  
In Vitro Plant Regeneration

Lychnis cognate Maxim and Lychnis fulgens Fish. Ex Spreng are two valued ornamental plants in Korea. Soaking of seeds in GA3 solution remarkably promoted germination up to 60%, but the control (0 mg/l) was not effective (> 5%). To select an adequate temperature for seed germination, seeds, previously soaked in a 1000 mg/l GA3 for 24 hrs, were incubated at 15, 20, 25, and 30°C. Seed germination of over 20% was obtained at 15, 20, and 25°C, but only 10% at 30°C. These results indicate that the seeds of L. cognate and L. fulgens are in a such dormant state that they hardly germinate even by dormancy breaker (GA3) and low (15 ? 25°C) temperature treatment. The highest callus induction was observed in the leaf explants of the seedlings on MS containing specific concentrations of 3.0 mg/l BA and 1.0 mg/l NAA. The adventitious shoot was formed < 90% of calli on 1/2 WPM medium. The height of in vitro propagated plantlet was no different media used for regeneration. This in vitro propagation protocol should be useful for conservation of endangered and ornamental plant.Plant Tissue Cult. & Biotech. 25(1): 1-12, 2015 (June)

scholarly journals In vitro shoot regeneration of boldo from leaf explants

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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