In vitro multiplication and hardening of grapevine plants in aeriated media

In vitro cultures have widely been used in horticulture for rapid multiplication of new varieties and clones as well as to produce pathogen-free stock material. To improve efficient hardening and transfer in vitro grown grapevine plants were multiplied by cutting them into single-node internodes with the whole leaf. Microcuttings including the shoot tips were rooted in granulated perlite moisted with tapwater under sterile conditions. After 2-3 weeks the rooted microcuttings were supplied by nutrients and hardened by gradual opening and finally by complete removal of the lids of jars or plastic boxes used for growth. Using this method microcuttings of Vitis vinifera cvs. „Chardonnay", „Cabernet franc", „Riesling" and „Sauvignon blanc" and the rootstock varieties Vitis riparia x Vitis cinerea cv. „Barrier" and Vitis berlandieri x Vitis rupestris cv. „Richter 110" formed new roots and shoots and 100% of the tested plants survived the acclimatization procedure. Similar results were obtained when perlite was replaced with rockwool-, or pit-pot blocks. This method may highly increase the efficiency of producing pathogen-free propagating material and new transgenic lines.

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Efficient method for rapid multiplication of clean and healthy willow clones via in vitro propagation with broad genotype applicability

Canadian Journal of Forest Research ◽

10.1139/cjfr-2015-0055 ◽

2015 ◽

Vol 45 (11) ◽

pp. 1662-1667 ◽

Cited By ~ 2

Author(s):

Elena Palomo-Ríos ◽

William Macalpine ◽

Ian Shield ◽

Joanna Amey ◽

Cuma Karaoğlu ◽

...

Keyword(s):

Rapid Multiplication ◽

Renewable Biomass ◽

Wide Range ◽

International Transport ◽

Breeding Programmes ◽

New Varieties ◽

Tissue Culture Propagation ◽

Willow Breeding ◽

Free Status

Willow is a versatile crop with considerable potential as a source of renewable biomass for bioenergy. Although breeding new varieties takes less time compared with some other tree species, producing new willow varieties is still a slow, labour-intensive process, partly because clonally propagating the results of each cross is a bottleneck early in the breeding scheme. In this paper, we describe a facile, rapid method for the in vitro culture of a wide range of willow genotypes. We have developed a combination of media and methods for efficient tissue-culture propagation to rapidly multiply individual plants and simultaneously produce clean, stock germplasm applicable to a wide range of willow genotypes that can be phytosanitary tested to demonstrate their disease-free status. The micropropagation method described could generate in the order of 5000 viable, transplantable clones from a single plant in just 24 weeks and was used to produce phytosanitary tested breeding material for export to overcome restriction on the international transport of woody cuttings. This method could represent a valuable biotechnology adjunct to willow breeding programmes and could accommodate early selection via molecular or biochemical markers.

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Micropropagation of Stevia rebaudiana plants

Ciência Rural ◽

10.1590/0103-8478cr20181029 ◽

2020 ◽

Vol 50 (1) ◽

Author(s):

Marta Teresa Rokosa ◽

Danuta Kulpa

Keyword(s):

Plant Growth ◽

Plant Growth Regulators ◽

Growth Regulators ◽

Stevia Rebaudiana ◽

In Vitro Cultures ◽

Optimum Composition ◽

Ms Medium ◽

Single Node ◽

Stevia Rebaudiana Bertoni

ABSTRACT: The aim of the study was to develop optimum composition of plant growth regulators in media for the propagation and rooting of shoots of stevia (Stevia rebaudiana Bertoni) in in vitro cultures. Single-node shoot fragments obtained from plants propagated on MS medium were placed onto media supplemented with: BAP, 2iP and KIN at concentrations: 0.5, 1, 2 and 5 mg∙dm-3, whereas at the rooting stage with addition of: IAA, IBA and NAA at concentrations 1, 2, 4 and 8 mg∙dm-3. The highest number of shoots and leaves was reported for plants propagated on MS medium enriched with 0.5 mg∙dm-3 BAP. The greatest number of the longest roots was developed by stevia on the MS medium enriched with 1 mg∙dm-3 IAA.

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Semi-sterilized Tissue Culture for Rapid Propagation of Grapevines (Vitis vinifera L.) Using Immature Cuttings

HortScience ◽

10.21273/hortsci.49.7.949 ◽

2014 ◽

Vol 49 (7) ◽

pp. 949-954

Author(s):

Fucheng Shan ◽

Kevin Seaton

Keyword(s):

Tissue Culture ◽

Vitis Vinifera ◽

Rapid Expansion ◽

Vitis Vinifera L ◽

Single Node ◽

Skill Levels ◽

Rooting Medium ◽

New Varieties ◽

Novel Method

Rapid expansion of grapevine plantings in many parts of the world has led to increased demand for desirable planting stocks. In countries that rely on importing new varieties and have strict quarantine rules, such as Australia, vines need to stay under quarantine for ≈2 years before they are released, at which time there is very limited wood available. Hence, rapid expansion of propagating stock after release is the key to multiplying up new varieties. A novel method, referred to as Semi-sterilized Tissue Culture (SSTC) using immature single-node cuttings, was established and evaluated as a way of rapid expansion of grapevine (Vitis vinifera L.) planting stock. In the SSTC method, immature single-node cuttings were surface-sterilized using methylated spirits and then cultured in the root pulsing medium [1/2 Murashige and Skoog (MS) medium supplemented with 40 μM indole-3-butyric acid (IBA)] for 24 hours. They were then planted in sterilized aerobic rooting medium (sphagnum peat:coarse river sand:perlite = 0.5:1:2) and cultured in a tissue culture room for ≈4 weeks for root initiation and development. The rooted immature single-node cuttings were then transferred to normal propagation beds in a greenhouse and potted on for acclimatization. Tube stock generated by SSTC easily acclimatized with a 15 times higher root strike rate than cutting propagation. It also took at least 50% less time than fully sterilized micropropagation methods to produce planting stocks. The advantages of the SSTC method are that it can be conducted under semisterilized conditions, avoiding degeneration and bacterial contamination problems encountered in micropropagation methods. By removing the time-consuming steps of the explant establishment, proliferation, and maintenance in vitro, the propagation process was simplified compared with conventional sterile tissue culture procedures. The SSTC procedure removed the need for high operator skill levels, reducing expense and allowing easier commercial adoption.

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Optimizing Initiation and Maintenance of Vitis Embryogenic Cultures

HortScience ◽

10.21273/hortsci.44.5.1400 ◽

2009 ◽

Vol 44 (5) ◽

pp. 1400-1406 ◽

Cited By ~ 28

Author(s):

Sadanand A. Dhekney ◽

Zhijian T. Li ◽

Michael E. Compton ◽

Dennis J. Gray

Keyword(s):

Somatic Embryogenesis ◽

Culture Media ◽

Medium Composition ◽

Embryogenic Competence ◽

Embryogenic Cultures ◽

Vitis Riparia ◽

Vitis Rupestris ◽

In Vitro Micropropagation ◽

Embryogenic Response

Stamens and pistils from mature grapevines and leaves from in vitro micropropagation cultures were used to optimize parameters influencing somatic embryogenesis in Vitis. Embryogenic competence was dependent on species/variety, explant type and developmental stage, medium composition, and growth regulator concentration. Of varieties evaluated, a greater number produced embryogenic cultures from stamens and pistils (26) compared with leaves (six). Among the different stamen and pistil stages, Stage II and III explants produced the maximum embryogenic response regardless of genotype and medium composition. Of seven culture media tested, the highest embryogenic response was recorded from varieties cultured on MSI (18) and PIV (16) media. Experiments annually repeated over 3 to 10 years demonstrated reproducible results. Highly reliable protocols for somatic embryogenesis were obtained for 29 Vitis species and varieties, including 18 Vitis vinifera varieties, Vitis riparia, Vitis rupestris, Vitis champinii, and eight Vitis hybrids. Embryogenic cultures were maintained on X6 medium for a period of 6 months to 2 years depending on the variety and used in studies involving genetic transformation and transgenic plant regeneration.

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Micropropagation of Mountain Mulberry (Morus bombycis Koidz.) ‘Kenmochi’ on Cytokinin-Free Medium

Plants ◽

10.3390/plants9111533 ◽

2020 ◽

Vol 9 (11) ◽

pp. 1533

Author(s):

Wojciech Litwińczuk ◽

Beata Jacek

Keyword(s):

Good Alternative ◽

In Vitro Cultures ◽

Axillary Shoot ◽

Free Medium ◽

Single Node ◽

Potted Plants ◽

Adult Trees ◽

Axillary Branching

The aim of the study was to compare two methods of micropropagation of mulberry: single-node culture (“SNC”), and axillary-branching (“AxB”). The experiments were carried out on in vitro cultures for 6 successive passages. The “AxB” cultures were propagated on modified MS medium (+ 25% Ca2+ and Mg2+), supplemented with WPM vitamins, sucrose (30 g L−1), and BA (1.5 mg l-1). The “SNC” cultures were grown on cytokinin-free 1/2 MS (macro- and micronutrients) medium supplemented with WPM vitamins, IBA (0.05 mg l-1), and sucrose (15 g l-1). Both media (pH 5.8) were solidified with agar (7.0 g l-1). Initiation of in vitro cultures from explants taken from adult trees and young, potted plants was feasible on both media. Cultures were established from about 1 cm long nodal explants. Generally “SNC” cultures formed one well rooted, significantly longer axillary shoot with bigger leaves than “AxB” cultures, which developed significantly more shoots and big callus at the explant base. All shoots collected from “SNC” and “AxB” cultures rooted in vivo in peat mixture and developed into similar plantlets. The single-node method based on application of cytokinin-free medium is a good alternative for the axillary-branching method for micropropagation of mountain mulberry (Morus bombycis) ‘Kenmochi”’.

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Combined In Vitro and In Vivo Propagation for Rapid Multiplication of Grapevine cv. Arka Neelamani

HortScience ◽

10.21273/hortsci.36.6.1107 ◽

2001 ◽

Vol 36 (6) ◽

pp. 1107-1110 ◽

Cited By ~ 4

Author(s):

Pious Thomas ◽

John W. Schiefelbein

Keyword(s):

Shoot Growth ◽

Multiplication Rate ◽

Normal Pattern ◽

Single Node ◽

Rapid Multiplication ◽

Root And Shoot Growth ◽

Subsequent Propagation ◽

Sprout Growth

A novel combination of in vitro and in vivo approaches was employed to generate sufficient stock of an introduced grape (Vitis vinifera L.) cv. Arka Neelamani which significantly accelerated the multiplication rate. The in vitro part included induction of root and shoot growth in shoot tip and nodal microcuttings in MS medium containing 1 μm IAA, sequential pruning of shoots at 1, 1.5, and 2 months, leaving the basal one to two nodes, resulting in fresh sprouts on the stump, and use of remaining stumps for in vivo establishment. The in vivo part included acclimatization of in vitro rooted plantlets and stumps, use of single node cuttings from 1.5- to 2-month-old in vivo shoots for the subsequent propagation, and utilizing the fresh sprout growth from these cuttings and in vivo stumps for further propagation. Employing both in vitro and in vivo approaches, we achieved a multiplication rate unparalleled to the general micropropagation or conventional propagation and significant stock was obtained within 6 months of introducing the material. The in vivo plants exhibited adult characters like distichous phyllotaxy, three lobed leaves and normal pattern of tendril development within 2 months from planting.

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