Micropropagation of Oleander (Nerium oleander L.)

Successful propagation of Nerium oleander L. (oleander) was achieved by in vitro methods. Shoot cultures were initiated from seedlings of wild-growing plants and from shoot apices of adult plants belonging to the commercial cultivars Splendens Giganteum, Revanche, and Alsace. Axillary shoot breaking from shoot tips excised from these cultures required the presence of either 6-benzylaminopurine (BA) or thidiazuron (TDZ). The higher number of axillary shoots from juvenile material was obtained by culturing shoot tips from BA-pretreated material derived from seedlings on a modified Schenk and Hildebrandt medium (SHM) supplemented with BA or TDZ (average of 3.9 shoots per explant with a mean length of 10.4 mm) and when the media were supplemented with 8.8 μM TDZ (average of 3.5 shoots per explant with a mean length of 7.3 mm) or 4.4 μM BA (average of 3.3 shoots per explant with a mean length of 12.3 mm). Among cultivars, cv. Revanche showed best shoot proliferation rates, especially when explants were cultured on Woody Plant Medium (average of 3.2 shoots per explant with a mean length of 10.2 mm). Adventitious bud differentiation from oleander leaves is also reported. Leaves excised from seedling-derived shoot cultures responded better than those from adult plant-derived shoot cultures (40% versus 5%, respectively). Bud differentiation required the presence of TDZ in the SHM medium, although shoot development was only achieved on transference of explants to media without TDZ but supplemented with BA and indoleacetic acid (IAA) or BA, kinetin, and IAA. Axillary and adventitious shoots were easily rooted (99%) and successfully (95% to 100%) transferred to soil.

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Development of in vitro shoot cultures of strawberry (Fragaria × ananassa Duch.) ‘Senga Sengana’ and ‘Elsanta’ under the influence of high doses of gibberellic acid

Folia Horticulturae ◽

10.2478/fhort-2013-0137 ◽

2009 ◽

Vol 21 (2) ◽

pp. 43-52 ◽

Cited By ~ 3

Author(s):

Wojciech Litwińczuk ◽

Ewa Okołotkiewicz ◽

Iwona Matyaszek

Keyword(s):

Gibberellic Acid ◽

Adventitious Shoots ◽

Shoot Culture ◽

Shoot Elongation ◽

Axillary Shoot ◽

Shoot Cultures ◽

Axillary Shoots ◽

High Doses ◽

Culture Growth

Abstract In this study, the influence of gibberellic acid (GA3) on strawberry in vitro shoot culture growth and development was investigated. ‘Senga Sengana’ and ‘Elsanta’ clones were grown on the medium recommended by Boxus (1999), supplemented with BA (0.5 mg dm-3), IBA (0.1 mg dm-3), glucose (40.0 g dm-3) and GA3 (0.1 - control, 1.0, 2.0, 5.0, 10.0 mg dm-3). In general, gibberellic acid improved axillary shoot elongation and reduced the growth of callus as well as the formation of roots and the development of adventitious shoots. GA3 applied at a concentration of 1.0-2.0 mg dm-3 significantly increased the number of axillary shoots (mainly crown shoots), whereas under higher (5.0-10.0 mg dm-3) doses it stimulated the development of runners. It seems that a new method of strawberry micropropagation based on the multiplication of axillary crowns and runners, which could also reduce the risk of domination of in vitro cultures with adventitious shoots, might be elaborated

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Vegetative Propagation of Phytophthora cinnamomi-Tolerant Holm Oak Genotypes by Axillary Budding and Somatic Embryogenesis

Forests ◽

10.3390/f11080841 ◽

2020 ◽

Vol 11 (8) ◽

pp. 841

Author(s):

Maria Teresa Martínez ◽

Francisco Javier Vieitez ◽

Alejandro Solla ◽

Raúl Tapias ◽

Noelia Ramírez-Martín ◽

...

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Phytophthora Cinnamomi ◽

Shoot Proliferation ◽

Holm Oak ◽

Induction Medium ◽

Axillary Shoot ◽

Oak Decline ◽

Shoot Cultures

Holm oak (Quercus ilex) is one of the most widely distributed tree species in the Mediterranean basin. High mortality rates have been observed in holm oak populations in the southwest of the Iberian Peninsula as a result of oak decline syndrome. Selection and propagation of genotypes tolerant to this syndrome could aid the restoration of affected areas. In this article, we report micropropagation and conservation procedures based on axillary budding and somatic embryogenesis (SE) of holm oak plants, selected for their tolerance to Phytophthora cinnamomi—the main biotic factor responsible for oak decline. Forced shoots were obtained from potted plants of eight different genotypes, and used as stock material to establish in vitro shoot proliferation cultures. Reliable shoot proliferation was obtained in seven out the eight genotypes established in vitro, whereas multiplication rates were genotype-dependent. The highest rooting rates were obtained by culturing shoots for 24 h or 48 h on rooting induction medium containing 25 mg L−1 indole-3-butyric acid, followed by transfer to medium supplemented with 20 µM silver thiosulphate. Axillary shoot cultures can be successful conserved by cold storage for 12 months at 4 °C under dim lighting. Shoot tips, excised from axillary shoot cultures established from tolerant plants, were used as initial explants to induce SE. Somatic embryos and/or nodular embryogenic structures were obtained on induction medium with or without indole-acetic acid 4 mg L−1, in two out the three genotypes evaluated, and induction rates ranged between 2 and 4%. Plantlet recovery was 45% after two months cold stratification of somatic embryos and eight weeks of culture on germination medium. Vegetative propagation of P. cinnamomi-tolerant Q. ilex trees is a valuable milestone towards the restoration of disease-affected areas.

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High-efficiency Propagation of Mature ‘Washington Navel’ Orange and Juvenile ‘Carrizo’ Citrange Using Axillary Shoot Proliferation

HortTechnology ◽

10.21273/horttech.26.3.278 ◽

2016 ◽

Vol 26 (3) ◽

pp. 278-286 ◽

Cited By ~ 1

Author(s):

Maria Luiza De Oliveira ◽

James G. Thomson ◽

Ed Stover

Keyword(s):

Root Length ◽

Shoot Proliferation ◽

Axillary Shoot ◽

Navel Orange ◽

Axillary Shoots ◽

Axillary Shoot Proliferation ◽

Full Strength ◽

Carrizo Citrange ◽

Washington Navel

In vitro axillary shoot proliferation can be used to increase availability of citrus (Citrus) types in high demand, while limiting somaclonal variation. However, established protocols could be improved to increase efficiency. Therefore, this study investigated some factors [plant growth regulators (PGRs), basal media, and successive subculturing] which affect the in vitro axillary shoot proliferation of mature ‘Washington Navel’ orange (Citrus sinensis) and juvenile ‘Carrizo’ citrange (C. sinensis × Poncirus trifoliata). In ‘Washington Navel’ orange, maximum axillary shoot induction (66.9% explants producing axillary shoots with a mean of 2.45 shoots per explant) was obtained in Driver and Kuniyuki walnut (DKW) medium supplemented with 0.1 mg·L−1 6-benzylaminopurine (BA), 0.05 mg·L−1 naphthalene acetic acid (NAA) along with 1 mg·L−1 6-furfurylaminopurine [kinetin (kin)], whereas in ‘Carrizo’ citrange, axillary shoot production was greatest (82.6% and 87.5% of explants producing axillary shoots with a mean of 4.3 and 4.1 shoots per explant) at 1.0 or 2.0 mg·L−1 BA in DKW medium. The initial nodal propagules (with basal tissue remaining from removed shoots) were repeatedly subcultured for six times every 4 weeks onto DKW medium with the same levels of PGRs used for initial culturing. Woody plant medium (WPM), Murashige and Skoog medium (MS), and DKW were also compared for rooting at quarter to full strength for salt components, all amended with 2.0 mg·L−1 indolebutyric acid (IBA) and 0.5 mg·L−1 NAA. MS at full strength provided the highest rooting in ‘Carrizo’ citrange (93%) and longest root length (58 mm), whereas half-strength MS provided the highest rooting in ‘Washington Navel’ orange (60% to 61%) and the longest roots (26 mm). Addition of 1 μm spermidine to the rooting medium enhanced root length only for ‘Washington Navel’ orange on full-strength MS, but accelerated rooting for both cultivars on all media. The plantlets were successfully transferred to greenhouse conditions, exhibiting normal development, with high uniformity, and no evidence of somaclonal variation.

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Comparative Rooting of Shoot Tips of Four Japanese Persimmon Cultivars vs. Shoots Regenerated from Roots Cultured In Vitro

HortScience ◽

10.21273/hortsci.35.5.940 ◽

2000 ◽

Vol 35 (5) ◽

pp. 940-944 ◽

Cited By ~ 4

Author(s):

Takuya Tetsumura ◽

Hisajiro Yukinaga

Keyword(s):

Histological Study ◽

Adventitious Shoots ◽

Shoot Proliferation ◽

Shoot Tips ◽

Diospyros Kaki ◽

Japanese Persimmon ◽

Rooting Ability ◽

Regenerated Plants ◽

In Vitro Roots

When cultured in vitro, roots of four Japanese persimmon (Diospyros kaki L.) cultivars formed adventitious shoots on MS medium with 10 μm zeatin and 0.01 μm indole-3-acetic acid, although their organogenetic capacities varied. Histological study revealed that the origin of the adventitious shoots was the pericycle. The regenerated shoots grew well on the shoot proliferation medium (MS with 5 μm zeatin). Final rooting percentages of shoots regenerated from roots of three of the four cultivars were greater than those of shoots that originated from shoot tips and that had been subcultured >50 times. Shoots regenerated from `Jiro' roots rooted 10 days earlier, had more roots than those from shoot tips, and maintained higher rooting ability over ten subcultures. Rooted `Hiratanenashi' shoots regenerated from roots survived better after acclimatization than those from shoot tips. No obvious variants were observed either in vitro or in the field. The trees regenerated from roots flowered within 4 years. These findings suggest that partial rather than true rejuvenation was responsible for both the early flowering and the juvenile characteristics, i.e., the enhanced rooting ability, observed in the regenerated plants. Chemical name used: 6-(4-hydroxy-3-methylbut-2-enylamino) purine (zeatin).

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In vitro propagation of pointed gourd (Trichosanthes dioica Roxb.) through encapsulated shoot tips

Bangladesh Journal of Agricultural Research ◽

10.3329/bjar.v34i4.5832 ◽

1970 ◽

Vol 34 (4) ◽

pp. 555-563 ◽

Cited By ~ 1

Author(s):

MA Malek

Keyword(s):

Sodium Alginate ◽

Basal Medium ◽

Shoot Proliferation ◽

Alginate Beads ◽

Multiple Shoot ◽

Shoot Tips ◽

Shoot Cultures ◽

Artificial Seed ◽

Pointed Gourd

Plants were regenerated from encapsulated shoot tips of pointed gourd. Shoot tips isolated from multiple shoot cultures of AM-8 and AM-15 cultivars of pointed gourd were encapsulated in sodium alginate beads. For germination and shoot proliferation, encapsulated shoot tips (artificial seed) were cultured in MS basal medium containing different concentrations and combinations of BAP and NAA. Use of MS medium resulted in 90% conversion of encapsulated shoot tips into plantlets. The results exhibited that BAP and combinations of BAP and NAA play an important role in germination of artificial seed being encapsulated by sodium alginate beads. The plantlets were successfully established in earthen pot. Under the present study, limited experimental efforts have been made to establish the protocol for encapsulating the shoot tips for the production of artificial seed and their subsequent regeneration. It is the first report in Bangladesh in developing artificial seed production technique using vegetative tissue of pointed gourd. Key Words: In vitro propagation; pointed gourd; shoot tips. DOI: 10.3329/bjar.v34i4.5832Bangladesh J. Agril. Res. 34(4) : 555-563, December 2009

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A Laboratory Exercise for Axillary Shoot Proliferation using Ajuga reptans

HortTechnology ◽

10.21273/horttech.4.3.312b ◽

1994 ◽

Vol 4 (3) ◽

pp. 312b-314 ◽

Cited By ~ 3

Author(s):

John E. Preece ◽

Carl A. Huetteman

Keyword(s):

Adventitious Shoots ◽

Shoot Proliferation ◽

Tween 20 ◽

Axillary Shoot ◽

Axillary Shoots ◽

Single Node ◽

Axillary Shoot Proliferation ◽

Laboratory Exercise ◽

Short Time ◽

Shoot Tip Explants

This exercise was developed for a plant propagation course to demonstrate, in a short time, the four stages of micropropagation, the effects of cytokinin concentrations, and the differences between adventitious and axillary shoots. Greenhouse-grown stock plants were brought into the laboratory, and 4- to 5-cm-long tips of runners were surface-dis-infested for 15 min in 0.5% NaClO with 1 ml of Tween 20/liter, followed by two 5-min rinses in sterile water. Working in the open laboratory near the bases of pairs of lit Bunsen burners, students placed either single-node or shoot tip explants (2 cm long, five replications) onto MS medium with 0, 1, or 10 μM BA. Cultures were in-cubated in parafilm-sealed culture tubes on open laboratory benches. Axillary shoots grew regardless of concentration of BA, and explants on medium with 10 μM BA produced the most callus and adventitious shoots. Microshoots were rooted and ac-climatized under mist in the greenhouse. This exercise can be performed in an open laboratory without the use of laminar flow hoods, specialized sterilizing equipment, or supplemental lighting.

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In vitro Shoot Multiplication of Bupleurum disticho-phyllum Wight - A Native Medicinal Plant of Southern India

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v17i2.2574 ◽

1970 ◽

Vol 17 (2) ◽

pp. 115-124 ◽

Cited By ~ 4

Author(s):

S. Karuppusamy ◽

T. Pullaiah

Keyword(s):

Medicinal Plant ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Shoot Formation ◽

Shoot Tip ◽

Axillary Shoot ◽

Axillary Shoots ◽

Axillary Shoot Proliferation ◽

Shoot Tip Explants

Shoot multiplication of Bupleurum distichophyllum was achieved from the nodal and shoot tip explants of mature plants using MS with different concentrations and combinations of growth regulators. Maximum explant response was from axillary shoots and the highest number of shoots per explant was obtained on MS fortified with 1.0 mg/l BAP. The highest degree of axillary shoot proliferation was found to be 74 and 70% for nodal- and shoot tip explants, respectively on the medium containing 1.0 mg/l BAP + 0.1 mg/l NAA. The combination of BAP and GA3 was also found to be effective for both type of explants. The degree of shoot formation was affected by explant types and the exogenous hormonal regime in the medium. The regenerated shoots were successfully rooted on MS supplemented with 2.0 mg/l IBA, after sequential hardening, survival rate was 71%. Key words: Bupleurum distichophyllum, Medicinal plant, Micropropagation, Conservation Plant Tissue Cult. & Biotech. 17(2): 115-124, 2007 (December) DOI: 10.3329/ptcb.v17i2.2574

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Micropropagation of Chinquapin (Castanea henryi) Using Axillary Shoots and Cotyledonary Nodes

HortScience ◽

10.21273/hortsci13292-18 ◽

2018 ◽

Vol 53 (10) ◽

pp. 1482-1486 ◽

Cited By ~ 2

Author(s):

Huan Xiong ◽

He Sun ◽

Feng Zou ◽

Xiaoming Fan ◽

Genhua Niu ◽

...

Keyword(s):

Adventitious Shoots ◽

Shoot Proliferation ◽

Shoot Multiplication ◽

Nodal Segments ◽

Direct Organogenesis ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Axillary Shoots ◽

Cotyledonary Nodes

Castanea henryi is an important woody grain tree species native to China. The objective of the current study was to find the suitable plant growth regulators (PGRs) and the optimal concentrations for direct organogenesis by using axillary shoots and cotyledonary nodes. Seeds were collected from the field, sterilized, and germinated in vitro. Axillary shoots and cotyledonary nodes of 3-week-old seedlings were used as explants. To find the suitable PGR for adventitious shoot induction, 0.5 mg·L–1 6-benzylaminopurine (6-BA), 0.1 mg·L–1 indole-3-acetic acid (IAA), 0.1 mg·L–1 2,4-dichlorophenoxyacetic acid (2,4-D), or 0.1 mg·L–1 1-naphthaleneacetic acid (NAA) was supplemented to Murashige and Skoog (MS) medium containing 0.65% agar and 3% sucrose. A high induction percentage of adventitious shoots (85.67%) was obtained from cotyledonary nodes supplemented with 0.1 mg·L–1 2,4-D. The type of explant influenced shoot proliferation rates and quality. Apical explants produced more and longer shoots than nodal segments. For shoot multiplication, 1 mg·L–1 6-BA + 0.05 mg·L–1 indole-3-butyric acid (IBA) supplemented with MS medium produced 12.33 and 6.25 shoots per explant, respectively, from apical and nodal explants. For shoot elongation and strengthening, 2 mg·L–1 6-BA + 0.05 mg·L–1 IBA supplemented with MS medium was the best combination, producing shoots with a mean length of 3.50 cm, a diameter of 0.46 cm, and about eight leaves per shoot. The greatest rooting of 76.70% and 11.33 roots per shoot was achieved when cultured in MS medium supplemented with 3.5% perlite + 1.5 mg·L–1 IBA. For acclimatization of the rooted plantlets in the greenhouse, a survival rate of 80% was achieved. This protocol—from multiplication to acclimation—is helpful to realize mass propagation of high-quality trees of chinquapin for increasing production and nut quality.

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Micro- and Cutting Propagation of Silver Maple. I. Results with Adult and Juvenile Propagules

Journal of the American Society for Horticultural Science ◽

10.21273/jashs.116.1.142 ◽

1991 ◽

Vol 116 (1) ◽

pp. 142-148 ◽

Cited By ~ 51

Author(s):

John E. Preece ◽

Carl A. Huetteman ◽

W. Clark Ashby ◽

Paul L. Roth

Keyword(s):

Shoot Proliferation ◽

Ethanol Solution ◽

Axillary Shoot ◽

Stem Cuttings ◽

Axillary Shoots ◽

Axillary Shoot Proliferation ◽

Silver Maple ◽

Adult Trees ◽

Juvenile Explants

Clonal micropropagation studies with silver maple (Acer saccharinum L.) included experiments with various shoot. explant types, cytokinins, and stock plant maturation levels. These trials led to successful explant establishment, axillary shoot proliferation, rooting of microshoots, and establishment of plantlets in the greenhouse. Overall, the best cytokinin tested was the phenylurea derivative TDZ. Shoot proliferation on juvenile explants was poor with kinetin, 2iP, and BA. Only zeatin at 10 μm was comparable to TDZ. TDZ at 10 nm was optimal for both juvenile and adult nodal explants. Juvenile explants that were held in vitro for 4 months commonly had at least 60 axillary shoots that could be subculture or excised for rooting. Microshoots rooted within 2 weeks. Following rooting, silver maple plantlets could be transplanted into a growing medium and placed directly onto a greenhouse bench. Studies were also conducted on rooting stem cuttings (macropropagation). Single nodes from juvenile plants rooted under intermittent mist, regardless of auxin application; however, shoot-tip cuttings from adult trees rooted best when auxin in ethanol solution was applied. Chemical names used: N- phenyl- N' -1,2,3 -thiadiazol-5-ylurea (thidiazuron, TDZ), N- (2-furanylmethyl)-1H-purin-6-amine (kinetin), isopentenyladenine (2iP), benzyladenine (BA), (E)-2-methyl-4-(1H-purin-6-ylamino)-2-buten-1-ol (zeatin).

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In vitro propagation of lnula royleana DC

Acta Societatis Botanicorum Poloniae ◽

10.5586/asbp.2004.001 ◽

2011 ◽

Vol 73 (1) ◽

pp. 5-8 ◽

Cited By ~ 3

Author(s):

Anna Stojakowska ◽

Janusz Malarz

Keyword(s):

In Vitro Propagation ◽

Shoot Proliferation ◽

Cotyledonary Node ◽

Axillary Shoot ◽

Ms Medium ◽

Axillary Shoots ◽

Axillary Shoot Proliferation ◽

Primary Explants ◽

Cotyledonary Node Explants

A micropropagation method, through axillary shoot proliferation, was elaborated for <em>Inula royleana </em>DC. (Asteraceae), a medicinal plant native of Himalaya. Primary explants (cotyledonary node explants) and secondary explants (node explants of in vitro regenerated shoots) of the plant, inoculated on MS medium supplemented with 0.1 μM NAA and 5.0 μM kinetin, regenerated 3.4 ± 1.2 and 5.1 ± 1.9 axillary shoots per explant, respectively. The regenerated shoots were easily rooting and adapting to growth in soil.

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