Clonal propagation of Pelargonium x hortorum through tissue culture: Effects of salt dilution and growth regulator concentration

1993 ◽  
Vol 73 (3) ◽  
pp. 871-878 ◽  
Author(s):  
Hélène Desilets ◽  
Yves Desjardins ◽  
Richard R. Bélanger
Keyword(s):  
Growth Regulators ◽  
Doubling Time ◽  
Culture Media ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Naphthaleneacetic Acid ◽  
High Survival ◽  
Half Strength

Different culture media were compared at the initiation and multiplication steps to develop a rapid production system for geranium (Pelargonium × hortorum) in vitro. Different salt dilutions of the Murashige and Skoog (MS) (1962) mineral medium were used in combination with different concentrations of 1-naphthaleneacetic acid (NAA) and 6-benzyladenine (BA) in order to optimize initiation of shoots of four geranium cultivars. The use of a MS basal medium with half-strength macrosalts supplemented with 0.11 μM NAA and 0.89 μM BA gave the best results in initiation. More than 40% of the apices initiated on this medium produced multiple shoots within a month. Subsequently, the effect of different concentrations of growth regulators was quantified by the mean of "shoot doubling time" evaluation. The shortest time recorded was 10.5 d for a theoretical production of 1 × 109 plantlets/apex/year. This is the first quantitative evaluation of geranium production in vitro. Geranium plantlets rooted easily on a half-strength MS medium without growth regulators. Acclimatization of geranium plantlets was characterized by high survival rates (94%) and the plants thus produced were phenotypically comparable to seed-derived plants. Key words: Geranium, micropropagation, shoot doubling time, in vitro

scholarly journals Propagation and Establishment of Three Endangered Mexican Orchids from Protocorms

HortScience ◽  
2009 ◽  
Vol 44 (5) ◽  
pp. 1395-1399 ◽  
Author(s):  
Martín Mata-Rosas ◽  
Víctor M. Salazar-Rojas
Keyword(s):  
In Vitro Propagation ◽  
Culture Media ◽  
Survival Rates ◽  
Basal Medium ◽  
Shoot Formation ◽  
Naphthaleneacetic Acid ◽  
Ex Vitro ◽  
Shoot Production ◽  
Further Development

Protocols for in vitro propagation from protocorms of Mormodes tuxtlensis Salazar, Cuitlauzina pendula La Llave & Lex., and Lycaste skinneri (Batem. Ex. Lind.) Lind., three endangered species distributed in Mexico and highly appreciated as ornamentals, were developed. The effect of two different culture media, Murashige and Skoog (MS) and modified Knudson (KCm), combined with varying concentrations of N6-benzyladenine (0, 2.2, 4.4, 8.9, and 13.3 μM) and α-naphthaleneacetic acid (0, 0.5 and 2.7 μM), were investigated. Shoot formation and development of protocorm-like bodies were observed. For all three species, cultures in MS produced more shoots per explant than those in KCm, and those shoots were longer and more robust in appearance. Maximum number of shoots for M. tuxtlensis (1.5) and C. pendula (24.3) were obtained in media supplemented with 13.3 μM and 2.2 μM N6-benzyladenine, respectively. Conversely, for L. skinneri the greatest shoot production (16.4) was achieved in medium supplemented with 2.7 μM α-naphthaleneacetic acid. Subculturing explants in MS basal medium allowed further development and rooting of the shoots as well as growth of protocorm-like bodies. The effect of different potting mixes on ex vitro survival plantlets was also investigated; pine bark:oak charcoal:pumice (3:1:1) allowed the highest survival rates in all three species.

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

scholarly journals Micropropagation and in vitro conservation of Alcantarea nahoumii (Bromeliaceae), an endemic and endangered species of the Brazilian Atlantic Forest

2020 ◽  
Vol 42 ◽  
pp. e52940
Author(s):  
Simone Sacramento dos Santos Silva ◽  
Everton Hilo de Souza ◽  
Fernanda Vidigal Duarte Souza ◽  
Cristina Ferreira Nepomuceno ◽  
Maria Angélica Pereira de Carvalho Costa
Keyword(s):  
Atlantic Forest ◽  
Culture Media ◽  
In Vitro Conservation ◽  
Naphthaleneacetic Acid ◽  
High Survival ◽  
Multiplication Rate ◽  
Ms Medium ◽  
Mature Seeds ◽  
Stem Segments

Alcantarea nahoumii (Leme) J. R. Grant is a species native to the Atlantic Forest that stands out for ornamental purposes. The objective of this work was to evaluate the in vitro germination of A. nahoumii seeds and establish a micropropagation protocol for production of seedlings so as to minimize the effects of predatory extractivism and develop an in vitro conservation system. Mature seeds were disinfested, established in three culture media (MS, MS½ and MS⅓) and incubated at four temperatures (20, 25, 30 and 35ºC) in a germination chamber. In the micropropagation experiment, stem segments were introduced in MS medium supplemented with 0.5 μM of 1-naphthaleneacetic acid (NAA) and 0.0, 2.2, 4.4 and 6.6 μM of 6-benzylaminopurine (BAP). For the in vitro conservation, plantlets were established in MS or MS½ medium supplemented with 15 g L-1 or 30 g L-1 of sucrose. The plants were acclimated with commercial substrate. The highest seed germination percentages were promoted by temperature conditions of 20 and 25ºC, with MS culture medium. The highest multiplication rate of shoots was obtained from the treatment without addition of the growth regulator or when combined with 2.2 μM of BAP + 0.5 μM of NAA. The acclimation of the plants occurred with high survival rate. The species can be conserved in vitro under slow growth condition for 24 months when incubated in MS medium supplemented with 30 g L-1 of sucrose.

scholarly journals In vitro Multiple Shoot Regeneration from Petunia hybrida

2019 ◽  
Vol 7 (10) ◽  
pp. 1554
Author(s):  
Rebaz Rasul Habas ◽  
Musa Turker ◽  
Fethi Ahmet Ozdemir
Keyword(s):  
Petunia Hybrida ◽  
Survival Rates ◽  
Basal Medium ◽  
Multiple Shoot ◽  
Shoot Length ◽  
Peat Moss ◽  
Naphthaleneacetic Acid ◽  
Ms Medium ◽  
Shoot Number

An efficient plant regeneration protocol was developed from in vitro germinated seeds of Petunia hybrida an ornamentally important plant in the family Solanaceae. Shoot tip and node explants of Petunia hybrida were cultured on MS basal medium supplemented with different concentrations and combinations of Benzyl amino purine (BAP), 1-Naphthaleneacetic acid (NAA), Indole-3-butyric acid (IBA) and Gibberellic acid (GA3). The highest shoot length was obtained from MS medium supplemented with 1 mg/l BAP + 1 mg/l NAA. The highest shoot number (3 shoots/explant) were obtained from MS medium supplemented with 0.6 mg/l BAP + 0.5 mg/l IBA. The isolated shoots were transferred to MS basal medium supplemented with different concentrations of GA3 ranging from 0.05, 0.2, 0.5 and 1 mg/l for shoot elongation. The highest shoot length (5.75 cm) was recorded from the MS medium supplemented with 0.2 mg/l GA3 +0.2 mg/l BAP. Rooting of regenerated shoots were achieved on MS medium supplemented with 0.1-1 mg/1 IBA and NAA. The regenerated shoots with well developed roots were successfully acclimatized and established in pots containing sterilized peat moss and grown under laboratory conditions with 70% survival rates.

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 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.

In Vitro Reproductive Development of a Diploid Wild Species, Arachis duranensis1

1994 ◽  
Vol 21 (2) ◽  
pp. 139-143
Author(s):  
Q. L. Feng ◽  
H. E. Pattee ◽  
H. T. Stalker
Keyword(s):  
Growth Regulators ◽  
Early Stage ◽  
Basal Medium ◽  
Reproductive Development ◽  
Naphthaleneacetic Acid ◽  
Embryo Abortion ◽  
Culture Techniques ◽  
Combination Treatments ◽  
Four Levels

Abstract Embryo abortion at an early stage of reproductive development is a major impediment for introgressing germplasm from wild to cultivated species of Arachis by interspecific hybridization. Ovule and embryo culture techniques have been used to rescue aborting hybrid embryos, but increased efficiency and recovery of very young tissues are still needed. The objective of this study was to induce growth and differentiation of A. duranensis proembryos. Seven-, 10-, and 14-d-old peg tips were cultured on a modified basal medium containing MS and B5 media combinations with 16 combination treatments using three growth regulators—1-naphthaleneacetic acid, gibberellic acid, and 6-benzylaminopurine—each at four levels. The results showed that seeds could be obtained in vitro by peg tip culture of four- to 16-celled proembryos. The favorable concentration ranges of growth regulators for pod formation and embryo development were 0.5-2.0 mg/L NAA, 0.05-0.5 mg/L GA3, and 0.05-0.2 mg/L 6-BAP. Over all three selected ages of pegs, the three best combinations of growth regulators resulted in 4.8, 4.7, and 3.5% pod formation, respectively.

scholarly journals In vitro Regeneration of Grass Pea (Lathyrus sativus L.)

2016 ◽  
Vol 4 (2) ◽  
pp. 1-8 ◽  
Author(s):  
P Saha ◽  
M Afrin ◽  
AKM Mohiuddin ◽  
AM Shohael
Keyword(s):  
In Vitro Regeneration ◽  
Basal Medium ◽  
Multiple Shoots ◽  
Nodal Explants ◽  
Lathyrus Sativus ◽  
Naphthalene Acetic Acid ◽  
Grass Pea ◽  
Half Strength ◽  
Lathyrus Sativus L

In vitro regeneration protocol for grass pea (Lathyrus sativus L.) was optimized using different concentrations and combinations of growth regulators. Direct shoot regeneration obtained through shoot organogenesis from different explants of grass pea cultured on MS medium supplemented with Gamborg B5 vitamin containing 6-benzylaminopurine (BAP), Thidiazuron (TDZ) and ?-naphthalene acetic acid (NAA). Highest percentage of shoots were obtained at 4.0 mg/l of BAP on nodal explants. Stunted multiple shoots were developed from nodal explants while 1.5 mg/l TDZ was used. About 56% of direct shoots were also obtained, while the combination of BAP (4.0 mg/l) and NAA (0.5 mg/l) were used. Regenerated plantlets were rooted most effectively (40%) in rooting medium containing half strength of MS basal medium containing 1.0 mg/l NAA. Well rooted plantlets were further successfully acclimatized to ambient humidity level and grown in controlled environment until hardening.Jahangirnagar University J. Biol. Sci. 4(2): 1-8, 2015 (December)

scholarly journals Clonal Micropropagation of representatives of the genus Dactylorhiza Neck. ex Nevski

2021 ◽  
Vol 4 (46) ◽  
pp. 17-17
Author(s):  
Alexander Saakian ◽  
◽  
Keyword(s):  
Survival Rate ◽  
Growth Regulators ◽  
In Vitro Propagation ◽  
Culture Medium ◽  
Culture Media ◽  
Ms Medium ◽  
Organic Additives ◽  
Clonal Micropropagation ◽  
Half Strength

Abstract The aim of this study is to develop and improve methods of in vitro propagation of representatives of Dactylorhiza: D.baltica , D. fuchsii. For the study, we used protocorms obtained by the asymbiotic germination of seed during 90 days. It has been established that half-strength of Murashige and Skoog (1962) medium (½ MS) supplemented with 1-2 mg/l 6-Benzylaminopurine(6-BAP), potato puree (20g/l), and charcoal (1g/l) effectively influenced the development of protocorms, and seedlings formation in the studied species. The result of the study showed that the survival rate of protocorms was high in all experimental culture media, but in D. fuchsii it was better at a concentration 2mg/l of 6-BAP (95.4%), while in D. baltica it was high at 1mg/l (87.0%). The highest percentage of multiple protocorms (68%) and the formation of new secondary protocorms in D. fuchsii (5,5±0,3 units) were observed on a culture medium containing 2 mg/l 6-BAP. The highest percent of rooting of D. fuchsii protosoms (78%) and length of roots (0.9cm) observed in ½ MS medium without growth regulators. During the development of D. baltica protosoms, the culture medium of ½ MS containing 1 mg/l 6-BAP had the best effect on the number of roots (1.8±0.1root/protosom), while the medium supplemented with 2mg/l of 6-BAP contributed to the formation of a larger number of new secondary protocorms (3,2±0,1protocorm/unit). During the subsequent cultivation of protosoms of D. baltica on a culture medium containing 1 mg/l it was observed an increase in the height of shoots (4,8±0,3 см), and the length of roots (2,2±0,1 см), wherein the number of newly formed protocorms was higher by 30% on the medium supplemented with 2 mg/l 6-BAP. Keywords: DACTYLORHIZA BALTICA, DACTYLORHIZA FUCHSII, IN VITRO, PROTOCORMS, ORGANIC ADDITIVES

scholarly journals 027 MULTIPLICATION OF ROSE SPECIES IN VITRO

HortScience ◽  
1994 ◽  
Vol 29 (5) ◽  
pp. 431d-431
Author(s):  
Yan Ma ◽  
David H. Byrne ◽  
Jing Chen ◽  
Amanda Byrne
Keyword(s):  
Basal Medium ◽  
Shoot Proliferation ◽  
Shoot Multiplication ◽  
Multiple Shoots ◽  
Naphthalene Acetic Acid ◽  
Good Growth ◽  
Lateral Buds ◽  
Half Strength ◽  
The Media

Several rose species (Rosa rugosa, R. wichuraiana, R. setigera, R. laevigata, R. banksiae, R. roxburghii, R. odorata and hybrids) were employed to establish the appropriate nutrient media for shoot multiplication and root initiation of cultured shoots and to describe a procedure for the successful transfer to soil of plants obtained in vitro. Cultured shoot tips and lateral buds from different genotypes proliferated multiple shoots on a basal medium (MS salt, vitamins, glycine, sucrose and agar) supplemented with 0mg/l to 6mg/l 6-benzylamino purine (BA) and 0mg/l to 0.5 mg/l naphthalene acetic acid (NAA). Most rose species cultured in a modified MS medium supplemented with 2mg/l BA showed good growth and shoot proliferation. The buds nearest the apex exhibited the slowest rate of bud development. Root development was enhanced and shoot development inhibited by lowering the concentration of MS salts to quarter- and half-strength. With difficult-to-root species, rooting was improved by supplementing the media with auxin or giving them 3-7days of dark treatment.

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