scholarly journals 117 An In Vitro Regeneration System for Basil (Ocimum basilicum L.)

HortScience ◽  
1999 ◽  
Vol 34 (3) ◽  
pp. 461E-461
Author(s):  
Winthrop B. Phippen ◽  
James E. Simon
Keyword(s):  
In Vitro Regeneration ◽  
Morphological Characteristics ◽  
Leaf Explants ◽  
Basal Medium ◽  
Induction Medium ◽  
Shoot Induction ◽  
Regeneration System ◽  
Transformation Protocol ◽  
Regeneration Procedure

A plant regeneration protocol was successfully developed for basil (O. basilicum L.). Explants from 1-month-old seedlings yielded the highest frequency of regeneration of shoots (37%) with an average number of 3.6 shoots per explant. Calli and shoot induction were initiated on Murashige and Skoog (MS) basal medium supplemented with thidiazuron (TDZ) (4 mg/L) for ≈30 days. Shoot induction and development was achieved by refreshing the induction medium once after 14 days. The most morphogenetically responsive explants were basal leaf explants from the first fully expanded true leafs of greenhouse-grown basil seedlings. Developing shoots were then rooted on MS media in the dark without TDZ. Within 20 days, rooted plantlets were transferred and acclimatized under greenhouse conditions where they developed normal morphological characteristics. This is the first report of a successful in vitro regeneration system for basil through primary callus. The establishment of a reliable regeneration procedure is critical when developing a transformation protocol for enhancing the production of basil for insect and disease resistance and improved essential oil constituents.

scholarly journals In vitro regeneration protocol of Rauvolfia serpentina L.

2018 ◽  
Vol 53 (2) ◽  
pp. 133-138 ◽  
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

scholarly journals Adventitious Shoot Regeneration from In Vitro Leaf Explants of the Peach Rootstock Hansen 536

Plants ◽  
2020 ◽  
Vol 9 (6) ◽  
pp. 755
Author(s):  
Angela Ricci ◽  
Luca Capriotti ◽  
Bruno Mezzetti ◽  
Oriano Navacchi ◽  
Silvia Sabbadini
Keyword(s):  
In Vitro Regeneration ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Cultures ◽  
Efficient System ◽  
Regeneration Rate ◽  
Factors Affecting

In the present study, an efficient system for the in vitro regeneration of adventitious shoots from the peach rootstock Hansen 536 leaves has been established. Twenty regeneration media containing McCown Woody Plant Medium (WPM) as a basal salt supplemented with different concentrations and combinations of plant growth regulators (PGRs) were tested. Expanded leaves along with their petiole from 3-week-old elongated in vitro shoot cultures were used as starting explants. The highest regeneration rate (up to 53%) was obtained on WPM basal medium enriched with 15.5 μM N6-benzylaminopurine (BAP). The influences on leaf regeneration of the ethylene inhibitor silver thiosulphate (STS) and of different combinations of antibiotics added to the optimized regeneration medium were also investigated. The use of 10 μM STS or carbenicillin (238 μM) combined with cefotaxime (210 μM) significantly increased the average number of regenerating shoots per leaf compared to the control. In vitro shoots were finally elongated, rooted and successfully acclimatized in the greenhouse. The results achieved in this study advances the knowledge on factors affecting leaf organogenesis in Prunus spp., and the regeneration protocol described looks promising for the optimization of new genetic transformation procedures in Hansen 536 and other peach rootstocks and cultivars.

scholarly journals Plant Regeneration through Protocorm-like Bodies Induced from Nodal Explants of Syngonium podophyllum ‘White Butterfly’

HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2129-2133 ◽  
Author(s):  
Jin Cui ◽  
Juanxu Liu ◽  
Min Deng ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Shoot Culture ◽  
Nodal Explants ◽  
Induction Medium ◽  
Naphthalene Acetic Acid ◽  
Ex Vitro ◽  
Petiole Explants

Syngonium podophyllum ‘White Butterfly’, one of the most popular ornamental foliage plants, is propagated almost exclusively through in vitro shoot culture. Ex vitro rooting, however, has been associated with severe Myrothecium leaf spot (Myrothecium roridum Tode ex Fr.). The objective of this study was to establish a method for regenerating well-rooted plantlets before ex vitro transplanting. Leaf and petiole explants were cultured on a Murashige and Skoog (MS) basal medium supplemented with N-(2-chloro-4-pyridyl)-N′-phenylurea (CPPU), N-phenyl-N′-1,2,3-thiadiazol-5-ylurea (TDZ), 6-benzyladenine (BA), or N-isopentenylaminopurine (2iP) with α-naphthalene acetic acid (NAA) and 2,4-dichlorophenoxyacetic acid (2,4-D), respectively. Calli formed from leaf explants cultured on the basal medium supplemented with CPPU or TDZ with 2,4-D or with NAA as well as from petiole explants cultured on the medium supplemented with BA, CPPU, or TDZ with 2,4-D or NAA. The calli, however, failed to differentiate, and shoot organogenesis did not occur. Culture of nodal explants on the MS basal medium supplemented with 9.84 μm 2iP, 8.88 μm BA, 8.07 μm CPPU, or 9.08 μm TDZ with 2.26 μm 2,4-D resulted in the formation of protocorm-like bodies, adventitious shoots, and subsequently well-rooted plantlets. MS basal medium supplemented with 19.68 μm 2iP and 1.07 μm NAA resulted in the highest percentage (92.9%) of nodal explants producing protocorm-like bodies and an average of 16.9 well-rooted plantlets per nodal explant. Adventitious shoots were able to root in the initial induction medium, but better root development occurred after shoots with protocorm-like bodies were transferred onto MS basal medium supplemented with 9.84 μm 2iP and 2.69 μm NAA. Regenerated plantlets were stable and grew vigorously with 100% survival rates after ex vitro transplanting to a container substrate in a shaded greenhouse.

scholarly journals Development of an Efficient Plant Regeneration System for the Selenium-hyperaccumulator Astragalus racemosus and the Nonaccumulator Astragalus canadensis

HortScience ◽  
2008 ◽  
Vol 43 (7) ◽  
pp. 2138-2142 ◽  
Author(s):  
Chiu-Yueh Hung ◽  
Jiahua Xie
Keyword(s):  
Plant Regeneration ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Shoot Formation ◽  
Naphthaleneacetic Acid ◽  
Shoot Induction ◽  
Regeneration System ◽  
Rooting Medium ◽  
Significant Difference

A method of in vitro plant regeneration for both the selenium-hyperaccumulator Astragalus racemosus ‘Cream Milkvetch’ and the nonaccumulator Astragalus canadensis ‘Canadian Milkvetch’ was developed with two induction media, M1 and M2. The M1 and M2 contain Murashige and Skoog basal medium plus vitamins, 8.07 μm N-(2-chloro-4-pyridyl)-N′-phenylurea, 2.5% (w·v−1) sucrose, 0.7% (w·v−1) agar (pH 5.7), and 0.89 μm or 3.12 μm a-naphthaleneacetic acid, respectively. In vitro cultures were initiated on these two types of media with three types of explants: cotyledons, hypocotyls, and roots. More than 93% of cultured explants from both species could form calli or calli with shoots. With regard to shoot formation, A. canadensis could produce multiple shoots from all types of explants more efficiently than A. racemosus. The highest shoot induction was approximately three shoots per explant in A. racemosus, whereas A. canadensis could reach ≈10 shoots per explant. M1 could induce more shoots than M2 no matter what type of explant was used, but the overall induction rates were no significant difference. Among the three types of explants used, the cotyledons were the best explants for shoot induction in A. canadensis, whereas hypocotyls were the best in A. racemosus. In A. racemosus, shoots could also be obtained from calli on the rooting medium containing Murashige and Skoog basal plus vitamins, 2.84 μm indole-3 acetic acid, 2.5% (w·v−1) sucrose, and 0.7% (w·v−1) agar (pH 5.7). Approximately 43% of A. canadensis shoots and 19% of A. racemosus shoots could be rooted on the rooting medium.

scholarly journals In Vitro Shoot Regeneration and Polyploid Induction from Leaves of Hypericum Species

HortScience ◽  
2009 ◽  
Vol 44 (7) ◽  
pp. 1957-1961 ◽  
Author(s):  
Elisabeth M. Meyer ◽  
Darren H. Touchell ◽  
Thomas G. Ranney
Keyword(s):  
Shoot Regeneration ◽  
Chromosome Doubling ◽  
In Vitro Regeneration ◽  
Leaf Explants ◽  
Shoot Induction ◽  
Bluish Green ◽  
Spontaneous Chromosome ◽  
Ornamental Traits ◽  
In Vitro Shoot Regeneration

Hypericum L. H2003-004-016 is a complex hybrid among Hypericum frondosum Michx., Hypericum galioides Lam., and Hypericum kalmianum L. and exhibits valuable ornamental characteristics, including compact habit, bluish green foliage, and showy flowers. Inducing polyploidy may further enhance the ornamental traits of this hybrid and provide new opportunities for hybridizing with other naturally occurring polyploid Hypericum sp. In this study, in vitro shoot regeneration and treatment of regenerative callus with the dinitroaniline herbicide oryzalin (3,5-dinitro-N4,N4-dipropylsufanilamide) were investigated as a means of inducing allopolyploidy. First, in vitro regeneration was optimized for callus and shoot induction by culture of leaf explants on medium supplemented with benzylamino purine (BA) or meta-topolin (mT) at 5, 10, or 15 μM in combination with indoleacetic acid (IAA) at 0, 1.25, 2.5, or 5 μM. Both BA and mT treatments successfully induced regenerative callus and shoots. Multiple regression analysis estimated maximum regenerative callus (94%) and shoot induction (18 shoots per explant) in medium supplemented with 5 μM BA and 3.75 μM IAA. In the second part of the study, exposure of regenerative callus to oryzalin at 0, 7.5, 15, 30, 60, or 90 μM for durations of 3, 6, or 9 d was investigated for polyploid induction. There was no survival for any of the calli in the 60- or 90-μM oryzalin treatments, but calli subjected to the other treatments exhibited some survival and polyploid induction. Duration had no effect on callus survival or ploidy level, but oryzalin concentration was a significant factor in both. The greatest percentage (44%) of polyploids was induced with 30 μM oryzalin. Spontaneous chromosome doubling was observed in 8% of control explants receiving no oryzalin treatment.

scholarly journals SHOOT ORGANOGENESIS FROM CAMBIAL DISC EXPLANTS OF PEAR.

HortScience ◽  
1992 ◽  
Vol 27 (6) ◽  
pp. 696b-696
Author(s):  
Richard L. Bell
Keyword(s):  
Shoot Organogenesis ◽  
Adventitious Shoots ◽  
Leaf Explants ◽  
Basal Medium ◽  
Induction Medium ◽  
Absolute Requirement ◽  
Initial Medium ◽  
Free Media ◽  
Macro Nutrients

Discs of cambial tissue were excised from actively growing shoots of `Bartlett' pear, and explanted directly on regeneration induction media. The basal medium was 1/2 strength MS macro-nutrients, MS micro-nutrients and organics, 8 g/l agar, and 30 g/l sucrose. Phytohormone treatments consisted of a factorial design of NAA (0 and 5μM) and TDZ (1, 2, 3, 4, and 5μM). After 4 weeks incubation in the dark, the explants were transferred to auxin-free media with identical concentrations of TDZ. There was an absolute requirement for auxin in the induction medium, as all discs on auxin-free initial media died without callusing. Maximum shoot regeneration 4 weeks after transfer to expression media was obtained with an initial medium containing 5μM NAA and 3μM TDZ, from which 30% of the explants produced one or more adventitious shoots. This rate of regeneration is similar to that obtained in some experiments with in vitro leaf explants, and provides an alternative system for regeneration of pear.

scholarly journals Plant Regeneration of Hot Pepper (Capsicum annuum L.) and Expression of Mouse Adenosine Deaminase (ADA) Gene via Agrobacterium-mediated Transformation

HortScience ◽  
1996 ◽  
Vol 31 (4) ◽  
pp. 572c-572
Author(s):  
Hak-Tae Lim ◽  
Kei-youn Lee ◽  
Yeoung-Sook Yoo ◽  
Duck-Chun Yang
Keyword(s):  
Plant Regeneration ◽  
Capsicum Annuum ◽  
In Vitro Regeneration ◽  
Induction Medium ◽  
Foreign Gene ◽  
Hot Pepper ◽  
Shoot Induction ◽  
Regeneration Rate ◽  
Cotyledon Explants

Since in vitro regeneration and transformation systems in hot pepper (Capsicum annuum L.) have not been available, the application of new genetic manipulations has been limited. Here we report an efficient procedure to regenerate whole pepper plants and to generate transgenic plants expressing a foreign gene was established. High frequency of plant regeneration was observed when hypocotyl and cotyledon explants were cultured on MS/B5 medium supplemented with NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, NAA 0.05 mg·L–1 plus zeatin 2.0 mg·L–1, IBA 10.0 mg·L–1 plus BA 1.0 mg·L–1, IAA 0.02 mg·L–1 plus zeatin 3.0 mg·L–1. An addition of AgNO3 5–10 μm to these media improved the regeneration rate by about 10%. For plant transformation, hypocotyl and cotyledon explants of pepper were preconditioned on kanamycin-free shoot induction medium for 48 hours. Then, co-cultivation with Agrobacterium tumeaacience was done on the co-culture medium for 2 days. The explants were then blotted in sterile filter paper and placed on shoot induction and selection medium containing kanamycin sulfate (100 mg·L–1) and carbenicillin (500 mg·L–1). PCR showed that the introduced ADA gene was integrated and stably expressed in the regenerated plants. ADA enzyme activities were checked by spectrophotometric analysis.

scholarly journals An efficient in vitro regeneration system via callus from hypocotyl and root of Iris laevigata

2020 ◽  
Author(s):  
Fengyi Li ◽  
Lijuan Fan ◽  
Haijing Fu ◽  
Yujia Liu ◽  
Ling Wang
Keyword(s):  
In Vitro Regeneration ◽  
Genetic Material ◽  
Reproductive Capacity ◽  
Penicillin G ◽  
Induction Medium ◽  
Multiplication Rate ◽  
Regeneration System ◽  
Root Explants ◽  
2,4 Dichlorophenoxyacetic Acid

Abstract Background Iris laevigata is an ornamental plant with strong cold resistance. However, its low reproductive capacity limits its landscape applications. The I. laevigata wild genetic resources also need to be protected. In order to develop an effective regeneration system, the optimum agar concentration for the induction medium was determined. Two explants (hypocotyl and root) were then cultured on medium containing different concentrations of plant growth regulator (PGR). In addition, three antibiotics were evaluated for controlling endophyte contamination. Results The highest induction rate (75.00%) was obtained from hypocotyl explants on Murashige and Skoog salt mixture (MS) medium containing 6-benzylaminopurine (6-BA) 0.5 mg L-1 + 2,4-Dichlorophenoxyacetic acid (2,4-D) 1.0 mg L-1 + 1-naphthylacetic acid (NAA) 0.4 mg L-1. The medium containing 6-BA 0.5 mg L-1 + 2,4-D 0.5 mg L-1 + NAA 0.2 mg L-1 achieved the greatest multiplication rate (73.33%). Media containing indole-3-butyric acid (IBA) 0.5 mg L-1 + 6-BA 1.5 mg L-1 + NAA 1.0 mg L-1 achieved the highest differentiation rate (39.72%) for hypocotyl induced calli. Medium containing 6-BA 2.0 mg L-1 + NAA 0.4 mg L-1 + kinetin (KT) 1.0 mg L-1 resulted in the highest differentiation rate (49.52%) for root induced calli. One hundred mg L-1 penicillin G resulted in the optimal rate for reducing endophyte contamination. Conclusions The research determined the optimum PGR concentrations for inducting and multiplying I. laevigata calli from hypocotyl and root explants and a satisfactory means for control endophyte contamination. This research will result in the efficient and reliable reproduction of I. laevigata for landscape applications, genetic development of new Iris varieties, and preservation of the wild genetic material.

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 Direct Shoot Organogenesis in Murraya paniculata (L.) Jack: A Prerequisite for Genetic Transformation

HortScience ◽  
2013 ◽  
Vol 48 (7) ◽  
pp. 938-941 ◽  
Author(s):  
Xiuli Shen ◽  
Vladimir Orbović ◽  
Manjul Dutt ◽  
William S. Castle ◽  
Frederick G. Gmitter
Keyword(s):  
Genetic Transformation ◽  
Shoot Organogenesis ◽  
In Vitro Regeneration ◽  
Ms Medium ◽  
Shoot Induction ◽  
Regeneration System ◽  
Murraya Paniculata ◽  
Direct Shoot Organogenesis ◽  
Direct Shoot

An efficient in vitro regeneration system through direct shoot organogenesis was established for Murraya paniculata (L.) Jack (Orange Jessamine). Epicotyls, leaves, roots, and cotyledons from in vitro-germinated seedlings and several plant growth regulators (PGRs) were evaluated for their effects on plant regeneration. Longitudinally cut epicotyl segments were observed to be the optimal explants followed by uncut epicotyls (not longitudinally cut). Roots, leaves, and cotyledons were not suitable as explants as a result of little or no shoot induction. Adventitious shoot induction was enhanced by the addition of 6-benzyladenine (BA). The highest percentage of shoot induction (87%) and the greatest number of shoots per explant (12.7) occurred on Murashige and Skoog (MS) medium supplemented with 15 μM BA from longitudinally cut epicotyls followed by 5.2 shoots per explant from uncut epicotyls. Optimal concentration of gibberellic acid (GA3) for shoot elongation was observed to be 15 μM. Eighty-five percent of the regenerated shoots produced roots with an average of three roots per shoot on MS medium supplemented with 5 μM indole-3-butyric acid (IBA). Our protocol for direct shoot organogenesis can potentially lead to the development of a robust method for production of transgenic plants of M. paniculata through Agrobacterium-mediated genetic transformation.

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