scholarly journals 348 In Vitro Establishment and Growth of Bermudagrass, Buffalograss, Saltgrass, and Zoysiagrass

HortScience ◽  
2000 ◽  
Vol 35 (3) ◽  
pp. 452B-452
Author(s):  
Harrison Hughes ◽  
Leigh Towill
Keyword(s):  
Growth Regulators ◽  
Growth Regulator ◽  
Basal Medium ◽  
Tween 20 ◽  
Sterile Water ◽  
Antimicrobial Compound ◽  
Running Water ◽  
In Vitro Establishment

There are turfgrasses species that are clonally propagated; notably bermudagrass, buffalograss, and zoysiagrass. Some of the early cultivars of these species are no longer widely grown, and may eventually be lost if not preserved. In order to facilitate studies on the long-term cryopreservation of these species and specific lines of saltgrass, it is necessary to develop suitable micropropagation procedures. We have developed protocol for the isolation and establishment of clean cultures in vitro for all four species. A 1/2-strength MS basal medium with Nitsch & Nitsch vitamins, 5 mg/L of thiamine, 2 mg/L of glycine, 30 g of sucrose, 7 g of agar with varying growth regulators has been used. Explant materials are prewashed in the greenhouse prior to a 15- to 30-min soapy wash in the laboratory. After a 30- to 60-min rinse in running water, nodal sections are surface-disinfested in 10% bleach with Tween 20 for 15 min, followed by three sterile water rinses. This procedure, sometimes with PPM (a proprietary antimicrobial compound), results in 50% or greater clean cultures. Rapidly growing nodal sections work best and preferably those not established in soil. We have tested various growth regulator combinations and have found that 10 mg/L of BA results in proliferation of buffalograss and saltgrass. However, proliferation remains relatively slow, requiring 8 to 12 weeks to develop sufficiently for subculture. Although we have succeeded in obtaining clean cultures of bermudagrass and zoysiagrass, proliferation is minimal, Further research is ongoing to develop a proliferative system with these two species.

Somatic Embryogenesis and In vitro Plant Regeneration in Vigna trilobata (L.) Verd. - A Potential Pasture Legume

1970 ◽  
Vol 19 (1) ◽  
pp. 89-99
Author(s):  
K. Choudhary ◽  
M. Singh ◽  
M. S. Rathore ◽  
N. S. Shekhawat
Keyword(s):  
Somatic Embryogenesis ◽  
Growth Regulators ◽  
Somatic Embryos ◽  
Basal Medium ◽  
Long Term Study ◽  
Continuous Application ◽  
First Time ◽  
Pasture Legume

This long term study demonstrates for the first time that it is possible to propagate embryogenic Vigna trilobata and to subsequently initiate the differentiation of embryos into complete plantlets. Initiation of callus was possible on 2,4-D. Somatic embryos differentiated on modified MS basal nutrient medium with 1.0 mg/l  of 2,4-D and 0.5 mg/l  of Kn. Sustained cell division resulted in globular and heart shape stages of somatic embryos. Transfer of embryos on to a fresh modified MS basal medium with 0.5 mg/l of Kn and 0.5 mg/l of GA3 helped them to attain maturation and germination. However, the propagation of cells, as well as the differentiation of embryos, were inhibited by a continuous application of these growth regulators. For this reason, a long period on medium lacking these growth regulators was necessary before the differentiation of embryos occurred again. The consequences for improving the propagation of embryogenic cultures in Vigna species are discussed. Key words: Pasture  legume, Vigna trilobata, Globular, Heart shape, somatic embryogenesis D.O.I. 10.3329/ptcb.v19i1.4990 Plant Tissue Cult. & Biotech. 19(1): 89-99, 2009 (June)

scholarly journals Plant Regeneration of Pampas Grass from Immature Inflorescences Cultured in Vitro

HortScience ◽  
1992 ◽  
Vol 27 (7) ◽  
pp. 841-843 ◽  
Author(s):  
C.D. Robacker ◽  
W.L. Corley
Keyword(s):  
Acetic Acid ◽  
Plant Regeneration ◽  
Developmental Stage ◽  
Shoot Regeneration ◽  
Growth Regulators ◽  
Growth Regulator ◽  
Basal Medium ◽  
Cortaderia Selloana ◽  
Dichlorophenoxy Acetic Acid

A micropropagation system to obtain plants from inflorescences of pampas grass (Cortaderia selloana Schult. `Pumila') was developed. Factors examined included developmental stage of inflorescence cultured and growth regulator combinations and concentrations that support explant establishment, shoot regeneration, and rooting. Immature inflorescences ≈300 mm long formed many shoot primordia when initially cultured on Murashige and Skoog basal medium containing 4.5 μm 2,4-D and 8.9 μm BA and subcultured to medium with 0.4 μm 2,4-D and 4.4 μm BA. Thereafter, monthly transfer to a medium without growth regulators yielded about three shoots per tube per month for more than 6 months. Most shoots rooted spontaneously and were easily hardened to greenhouse conditions. Field-tested plants flowered within 2 years and nearly all appeared identical to the parent cultivar. With this technique, several thousand plants can be obtained from a single inflorescence in 1 year. Chemical names used: N -(phenylmethyl)-1 H -purine-6-amine (BA); (2,4-dichlorophenoxy)acetic acid (2,4-D).

scholarly journals In vitro organogenesis and plant regeneration of Passiflora xishuangbannaensis, a species with extremely small populations

2021 ◽  
Author(s):  
Yuan-yuan Meng ◽  
Shi-jie Song ◽  
Sven Landrein
Keyword(s):  
Plant Regeneration ◽  
Plant Growth ◽  
Plant Growth Regulators ◽  
Growth Regulators ◽  
Basal Medium ◽  
Ex Situ ◽  
Naphthaleneacetic Acid ◽  
South American ◽  
South American Species

Abstract Passiflora xishuangbannaensis (Passifloraceae) is endemic to a few sites of Mengyang nature reserve in Yunnan, Xishuangbanna and less than 40 individuals have been recorded. Nine Passiflora species are endemic to Yunnan with most species occurring in South America, making P. xishuangbannaensis highly significant and emblematic to the conservation work in the region. This study is designed to provide the first protocol for in vitro organogenesis and plant regeneration for ex situ conservation and reintroduction for an Asian Passiflora species. Using internodes, petioles and tendrils we optimize calli formation and root elongation using several plant growth regulators, individually or in combination. We also assess the genetic stability of regenerated cells. The maximum callus induction and shoot bud differentiation were both achieved on half Murashige and Skoog basal medium supplemented with 4.44 µM 6-Benzylaminopurine and 1.08 µM 1-Naphthaleneacetic acid. The best rooting was achieved from 30 days old, regenerated shoots on half Murashige and Skoog basal medium supplemented with 1.08 µM 1-Naphthaleneacetic acid. Micropropagated plants were subjected to inter simple sequence repeat markers analyses. Collectively, 86 bands were generated from 6 primers of which 12 bands were polymorphic, showing genetic variation between the regenerated plantlets and the original plant. Response to plant growth regulators was more specific than most other studies using South American species, which could be explained by the morphological and physiological differences between South American and Asian Passiflora species

2 BOVINE EMBRYONIC STEM-LIKE CELLS DERIVED FROM IN VITRO-PRODUCED BLASTOCYSTS

2017 ◽  
Vol 29 (1) ◽  
pp. 108
Author(s):  
Y. S. Bogliotti ◽  
J. Wu ◽  
M. Vilariño ◽  
K. Suzuki ◽  
J. C. Belmonte ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Cell Mass ◽  
Embryonic Stem ◽  
Basal Medium ◽  
Primitive Endoderm ◽  
Culture Dish ◽  
Rock Inhibitor ◽  
Immunodeficient Mice

Embryonic stem cells (ESC) are derived from the inner cell mass (ICM) of preimplantation blastocysts. To date, it has been challenging to establish pluripotent ESC lines for domestic animals, which could be important for biotechnological applications, such as genetic engineering and SCNT, and biomedical research. The aim of this work was to derive and characterise bovine embryonic stem-like cells (bESC) from in vitro-produced bovine blastocysts. Embryos were produced by in vitro fertilization of in vitro-matured oocytes aspirated from abattoir ovaries and cultured in groups of 25 in 50-μL drops of KSOM (Evolve, Zenith Biotech) with 4 mg mL−1 BSA for 7 days until they reached the blastocyst stage (Ross et al., 2009 Reproduction 137, 427–437). At that point, the zona pellucida (ZP) was removed using 1 mg mL−1 Pronase (Sigma, St. Louis, MO), and ZP-free blastocysts were washed 6 times in SOF-HEPES. Three derivation approaches were tested: ZP-free whole blastocysts, mechanically isolated ICM, and immunosurgery-derived ICM. In each case, individual blastocysts/ICM were placed in 1 well of a 12-well dish seeded with a monolayer of mouse embryo fibroblasts (MEF) and cultured in mTeSR1 basal medium (without growth factors) supplemented with 20 ng mL−1 FGF2 and 2.5 μM IWR1 (CTFR) (Wu et al. 2015 Nature 521, 316–321). After 48 h, blastocysts/ICM that failed to adhere were physically pressed against the bottom of the culture dish with a 22-gauge needle under a stereoscope to aid attachment. Thereafter, the media was changed daily. Outgrowths (after 6–7 days in culture) were dissociated and passaged using TrypLE and re-seeded in the presence of ROCK inhibitor (Y-27632, 10 μM) onto newly prepared wells containing MEF. Established bESC lines were cultured on MEF and passaged every 4 to 5 days at a 1:10 split ratio. The bESC lines were characterised by immunofluorescence (IF), RNA-seq, and teratoma formation. The efficiency of cell line derivation (evaluated at passage 3) was similar for the 3 approaches: whole blastocysts (9/16, 56.3%), mechanical ICM isolation (7/12, 58.3%), and immunosurgical ICM isolation (7/16, 43.8%). The bESC were passaged and cultured long-term (more than 15 passages) and were subjected to several rounds of freezing and thawing while retaining their morphology and characteristics. IF analysis showed that long-term cultured bESC expressed the markers SOX2 and OCT4 (pluripotency), but did not express CDX2 (trophectoderm) or GATA6 (primitive endoderm). RNAseq analysis of 2 bESC lines showed that ICM markers (POU5F1, NANOG, SOX2, LIN28B, DNAMT3B, UTF1, SALL4) were expressed (RPKM > 0.4), while trophectoderm markers (CDX2, GATA2, GATA3, FGF4, TFAP2A) and primitive endoderm markers (GATA6, HNF4A) were not expressed (RPKM < 0.4). Finally, bESC lines (n = 2) were able to form teratomas in immunodeficient mice. The teratomas contained tissues representative of the 3 germ lineages and expressed lineage-specific markers (ectoderm: TUJ1, endoderm: FOXA2, and mesoderm: ASM). In conclusion, the culture condition used in this work (CTFR) enables robust derivation and long-term in vitro propagation of pluripotent bESC.

scholarly journals In Vitro Propagation of Lilium longiflorum Bulbs Using NAA and BAP Plant Growth Regulator Treatment

2020 ◽  
Author(s):  
Ni Wayan Deswiniyanti ◽  
Ni Kadek Dwipayani Lestari
Keyword(s):  
Growth Regulators ◽  
Growth Regulator ◽  
In Vitro Propagation ◽  
Culture Media ◽  
Lilium Longiflorum ◽  
Herbaceous Plant ◽  
Perennial Herbaceous Plant ◽  
Plant Parts ◽  
Lily Bulb

Lily (Lilium longiflorum) is a perennial herbaceous plant with white trumpet-shaped flowers, fragrant and bulbous. In vitro culture through bulbs is one of way propagation of lily plants, but it requires a long time and only produces limited plants. In vitro propagation is a very promising technique for plant propagation because it can produce a lot of plant seeds in a short time. Bulbs are one of the fastest explants for growing shoots in lilies, but it is not known for certain which cuts of explants from bulb scales are best for multiplying in vitro. This study aims to determine the effect of lily bulb explants and the concentration of NAA and BAP growth regulators on the growth of lily bulb explants. The best results were obtained on the base and middle cuts explant of bulb scales compared to the tip cuts explant ones. The best results of the growing percentage, the number of shoots and the best growing time are shown in the combination treatment of growth regulator 1 mg L−1 NAA and 1 mg L−1 BAP. The optimum results on the number of micro bulbs were found in the treatment of growth regulators 0.5 mg L−1 NAA and 1 mg L−1 BAP. The best results of the average time formed micro bulb was in the treatment of 1 mg L−1 NAA and BAP with middle explant cuts, and treatment concentrations of 0.5 mg L−1 NAA and BAP in the base explant section. The base and middle bulb explants are able to regenerate or grow higher shoots. This is caused by the presence of endogenous natural auxin and the spread of auxin in plant parts not in the same amount. Therefore when added to the exogenous growth regulator such as auxin or cytokines to culture media will further trigger the formation of micro tubers more quickly,. It can increase the concentration of endogenous growth regulators in cells, help growing process and developing tissue.   Keywords: Bulb, lily, micro bulbs, in vitro, shoots

scholarly journals Use of growth regulators and experimental LED phytoirradiator in clonal micropropagation of garden strawberry (Fragaria × ananassa, Duchesne ex Weston)

2019 ◽  
Vol 20 (4) ◽  
pp. 324-333 ◽  
Author(s):  
M. G. Markova ◽  
E. N. Somova
Keyword(s):  
Experimental Data ◽  
Growth Regulators ◽  
Growth Regulator ◽  
Fragaria Ananassa ◽  
Clonal Micropropagation ◽  
Fluorescent Lamps ◽  
Combined Use ◽  
The Impact ◽  
Reproduction Factor

The article provides experimental data of 2017-2018 study on the effect of growth regulators and LED phytoirradiator on the proliferation and rooting of promising garden strawberry (Fragaria ananassa) varieties in vitro. Micro-shoots of Korona and Brighton strawberry varieties were taken as the object of the research. Strawberry micro-shoots were cultivated under fluorescent lamps in the control variant. A programmable combined blinking LED phytoirradiator was under study. The combined effect of cytokinin and gibberellic acid by adding them to the Murashige and Skoog nutrient medium, as well as the impact of Siliplant and EcoFus growth regulators on strawberry micropropagation has been studied. It was established that in the cultivation of Korona variety the combined use of Siliplant and EcoFus under illumination with LED phytoirradiator provided an increase in the reproduction factor. The coefficient was 5.0 pcs./explant that was 1.7 times higher than the control (3.0 pcs/explant), the LSD05 1.4 pcs/explant. The maximum reproduction factor of remontant strawberry Brighton variety was obtained in the variant with the use of Siliplant and LED phytoirradiator and amounted to 4.9 pcs./explant (4.2 pcs./explant in the control), the LSD05 was 1.5 pcs./ explant. Regardless of the lighting, the use of RibavExtra in all variants under study increased the rooting rate of the strawberry Korona micro-shoots from 92.8 to 99.1%, the LSD05 6.1%. The use of LED phytoirradiator in comparison with the luminescent one (94.3%) provided a significant increase in the rooting rate of the strawberry Korona micro-shoots to 98.1% regardless of the growth regulators used, the LSD05 3.5%. The combined use of LED phytoirradiator and Ribav-Extra growth regulator in concentrations of 1.0 and 1.5 mg/l resulted in rooting of strawberry Korona micro-shoots up to 100%. Regardless of the growth regulator used, the use of LED phytoirradiator in comparison with the luminescent one (88.9%) provided a significant increase in the rooting rate of the strawberry Brighton micro-shoots to 97.2%, the LSD05 4.6%. The rooting rate of the remontant strawberry Brighton microshoots was 100% in the variant with the use of Ribav-Extra in the concentration of 1.0 mg /l combined with LED phytoirradiator 20 days after transplanting for rooting.

scholarly journals THE EFFICIENCY OF SHOOT AND PLANTLET FORMATION OF Cephaelis ipecacuanha AFTER THREE SUBCULTURES IN VITRO

1994 ◽  
Vol 24 (3) ◽  
pp. 523-526 ◽  
Author(s):  
Osmar Alves Lameira ◽  
Marly Pedroso da Costa ◽  
José Eduardo Brasil Pereira Pinto
Keyword(s):  
Gibberellic Acid ◽  
Butyric Acid ◽  
Growth Regulators ◽  
Activated Charcoal ◽  
Basal Medium ◽  
Adventitious Shoot ◽  
Shoot Elongation ◽  
Plantlet Formation ◽  
Cephaelis Ipecacuanha

Multiple adventitious shoot formed from internodal segments of Cephaelis ipecacuanha cultured 25 days on Gamborg basal medium (GAMBORG et al., 1968) supplemented with 6.66mM 6-benzylaminopurine there was a maximum of nine shoots per segment and an average of five shoots per segment formed. The presence of gibberellic acid in the subculture media promoted shoot elongation in all treatments. The shoots attained 3cm in height and rooting of 100% after 35 days of culturing upon Murashige and Skoog's basal medium (MS), added with 4.92mM indole-3-butyric acid, 0.87m gibberellic acid and 0.1% activated charcoal. Further growth was accelerated after the transfer to 1/2 MS without growth regulators. Rooted plantlets transferred to potting soil could be successfully established.

scholarly journals USE OF FORCING SOLUTIONS TO STUDY PLANT GROWTH REGULATOR EFFECTS ON VANHOUTTE'S SPIRAEA CULTURED IN VITRO

HortScience ◽  
1990 ◽  
Vol 25 (9) ◽  
pp. 1124e-1124
Author(s):  
Guochen Yang ◽  
P. E. Read
Keyword(s):  
Plant Physiology ◽  
Plant Growth ◽  
Growth Regulators ◽  
Growth Regulator ◽  
Plant Growth Regulator ◽  
Woody Plant ◽  
Shoot Proliferation ◽  
Plant Tissues ◽  
Forcing Solution

Vanhoutte's spiraea has been propagated in vitro using explants from softwood growth of dormant stems forced in a solution containing 200 mg/l 8-hydroxyquinoline citrate (8-HQC) and 2% sucrose (Yang and Read, 1989). Objectives to further utilize this system were to determine the feasibility of applying plant growth regulators (PGR) via the forcing solution to softwood growth from forced dormant stems and to study the resulting influence on in vitro culture. BA and GA3 were placed in the forcing solution at various concentrations, including a zero PGR control. Explants were cultured on Linsmaier and Skoog (LS) medium containing zero PGR or different amounts of BA or thidiazuron (TDZ) or combinations of BA and IAA. Control explants placed on LS medium supplemented with 5uM BA with or without 1 or 5uM IAA, or with 0.5 or 0.75 uM TDZ alone produced the best shoot proliferation. BA in the forcing solution stimulated micropropagation, while GA3 caused less proliferation than explants from control solutions. Forcing solutions containing PGR are useful for manipulating responses of plant tissues cultured in vitro and for studying PGR influence on woody plant physiology.

scholarly journals A Highly Efficient In Vitro Cranberry Regeneration System Using Leaf Explants

HortScience ◽  
2000 ◽  
Vol 35 (5) ◽  
pp. 948-952 ◽  
Author(s):  
Luping Qu ◽  
James Polashock ◽  
Nicholi Vorsa
Keyword(s):  
Growth Regulators ◽  
Adventitious Shoots ◽  
Basal Medium ◽  
Naphthaleneacetic Acid ◽  
Leaf Position ◽  
Regeneration System ◽  
Abaxial Side ◽  
Adventitious Regeneration ◽  
Explant Orientation

A very efficient adventitious regeneration (shoot organogenesis) system for cranberry (Vaccinium macrocarpon Ait.) leaves was developed. A basal medium consisting of Anderson's rhododendron salts and Murashige and Skoog's (MS) organics, supplemented with 10.0 μm thidiazuron (TDZ) and 5.0 μm 2ip, was effective for adventitious regeneration from leaves for the five cranberry cultivars tested: `Early Black', `Pilgrim', `Stevens', `Ben Lear', and `No. 35'. Parameters examined included: 1) varying combinations of three plant growth regulators (TDZ, 2ip, and NAA); 2) explant orientation (adaxial vs. abaxial side in contact with the medium); and 3) leaf position relative to the apical meristem from the source plant. Cultivars varied in regeneration frequency, but cultivar × growth regulator interaction was nonsignificant. With optimal treatment conditions, regeneration occurred on more than 95% of the explants, with `Early Black' and `Pilgrim' producing as many as 100 shoot meristems per explant. At all concentrations tested, NAA (as low as 0.1 μm) increased callus formation and significantly reduced regeneration. Emerging adventitious shoots were always observed on the adaxial side of the leaves regardless of explant orientation on the medium. Regeneration was much greater when the abaxial side was in contact with the medium, and was not related to leaf position on the source plants. Elongation of adventitious shoots began ≈2 weeks after transfer to the basal medium without growth regulators. Cuttings of elongated shoots rooted 100% both in vitro in the basal medium and ex vitro in shredded sphagnum moss. The high regeneration efficiency achieved by using this system will be very useful in the application of techniques, such as Agrobacterium- and particle bombardment-mediated transformation. Chemical names used: 1-phenyl-3-(1,2,3-thiadiazol-5-yl) urea (thidiazuron, TDZ); N6-(γ-γ-dimethyallylamino) purine (2ip); α-naphthaleneacetic acid (NAA).

scholarly journals In Vitro Long-Term Cultures of Papaya (Carica Papaya L.).

2021 ◽  
Author(s):  
Jose Javier Regalado González ◽  
Manuel López Granero ◽  
Carlos Lopez Encina
Keyword(s):  
Morphological Characteristics ◽  
Basal Medium ◽  
Multiplication Rate ◽  
Ms Medium ◽  
High Quality ◽  
Poor Growth ◽  
Average Multiplication ◽  
Half Strength

Abstract We present the data on proliferation corresponding to 10 years of continuous incubation in vitro of papaya shoots, and propose a reliable method for long-term micropropagation for papaya, using two types of explants: Microshoots from somatic embryos, and from axillary buds of papaya. Three different media were assayed. The proliferation medium (PPRM) allowed to maintain papaya shoots under continuous proliferation during 20 years, maintaining a consistent behaviour. Most of the shoots developed in PPRM rooted during the incubation, and after acclimated easily, maintaining the morphological characteristics of the parental plants, flowering and setting fruits normally. The PPRM medium consist in MS medium supplemented with NAA (0.1 mg l-1), BA (0.5 mg l-1), GA3 (0.5 mg l-1) and Adenine sulphate (40 mg l-1). The average multiplication rate was higher than 20 shoots per explant along the long-term assay. The elongation medium (PELM), was designed to recover shoots with a poor growth, and allowed the development of high quality shoots ready for rooting, and consist in a MS basal medium supplemented with NAA (0.1 mg l-1), Kin (0.5 mg l-1) and GA3 (1 mg l-1). The rooting medium (PROM) was designed to induce high quality roots from non-rooted shoots and consist in a half strength MS medium plus IBA (1mg l-1). On PROM, agar can be exchanged for expanded vermiculite. Acclimation took place inside an acclimatization tunnel under progressive hydric stress. After 4 weeks, the plant recovery rate was 90% for plants maintained under continuous proliferation during ten years.

Sign in / Sign up

Export Citation Format

Share Document