Shoot Regeneration Rates of Perennial Phlox are Dependant on Cultivar and Explant Type

Meristem culture and/or thermotherapy were used for virus elimination from ornamental Phlox paniculata L. (`Blue Boy', `Orange perfection' and `Starfire') mother plants. Shoot tip, leaf, node and flower ovary explants collected from greenhouse-maintained virus free plants were cultured in vitro for shoot initiation. Adventitious shoot initiation was observed on Murashige and Skoog (MS) medium containing the cytokinin BA with or without the auxin NAA. The addition of 0.4 mg·L-1 thiamine, 0.4 mg·L-1 folic acid, and 40 mg·L-1 adenine sulfate to the MS medium did not improve the regeneration rate. Multiplication and rooting were genotype dependent. Blue Boy and Orange Perfection cultivars regenerated the maximum number of shoots from leaf explants. `Blue Boy' leaf explants from in vitro plants had a lower regeneration rate than explants from greenhouse plants. Cultivar `Starfire' had the highest shoot formation with open flower ovary explants and failed to regenerate from leaf explants. In vitro rooting of adventitious shoots in the presence of auxins (IAA, NAA, or IBA) with or without BA was less effective than ex vitro rooting. Chemical names used: 6-benzyladenine (BA); indole-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA).

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High Efficiency Direct Shoot Organogenesis from Leaf Segments ofAerva lanata(L.) Juss. Ex Schult by Using Thidiazuron

The Scientific World JOURNAL ◽

10.1155/2014/652919 ◽

2014 ◽

Vol 2014 ◽

pp. 1-6 ◽

Cited By ~ 5

Author(s):

K. Varutharaju ◽

C. Soundar Raju ◽

C. Thilip ◽

A. Aslam ◽

A. Shajahan

Keyword(s):

Shoot Organogenesis ◽

Adventitious Shoots ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Scale Production ◽

Ms Medium ◽

Leaf Segments ◽

Direct Shoot Organogenesis ◽

Direct Shoot

An efficient protocol for direct shoot organogenesis has been developed for the medicinal plantAerva lanata(L.) Juss. ex Schult. Regeneration was achieved from leaf segments of 20 days oldin vitroplantlets raised on Murashige and Skoog (MS) medium containing 0.25–2.0 mg L−1thiadiazuron (TDZ), 3% sucrose, and 0.8% agar. After 21 days of culture incubation, maximum number of shoot organogenesis (23.6 ± 0.16) was obtained on medium containing 1.0 mg L−1TDZ. The shoots were able to producein vitroflowers on medium containing 1.0 mg L−1TDZ in combination with 0.25–0.5 mg L−1 α-naphthaleneacetic acid (NAA). Histological observation showed that the epidermal cells of the leaf explants exhibited continuous cell division led to formation of numerous dome shaped meristematic protrusions and subsequently developed into adventitious shoots. Upon transfer of shootlets to half strength MS medium containing 1.0 mg L−1indole-3-butyric acid (IBA), around 86% of the regenerated shoots formed roots and plantlets. Rooted plants were hardened and successfully established in the soil at the survival rate of 92%. The regeneration protocol developed in this study provides an important method of micropropagation of this plant. Furthermore, this protocol may be used for a large scale production of its medicinally active compounds and genetic transformations for further improvement.

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Micropropagation of Dianthus gratianopolitanus

HortScience ◽

10.21273/hortsci.39.5.1083 ◽

2004 ◽

Vol 39 (5) ◽

pp. 1083-1087 ◽

Cited By ~ 4

Author(s):

Margarita Fraga ◽

Mertxe Alonso ◽

Philippe Ellul ◽

Marisé Borja

Keyword(s):

Indole Acetic Acid ◽

Adventitious Shoots ◽

Shoot Tip ◽

Naphthaleneacetic Acid ◽

Optimum Number ◽

Meristem Culture ◽

Axillary Shoots ◽

Shoot Initiation ◽

Mother Plants

Meristem culture and/or thermotherapy were used to eliminate viruses from ornamental Dianthus gratianopolitanus Vill. (`Spotti' and `Frosty Fire') mother plants. Shoot tip, leaf, node, and ovary explants collected from greenhouse-maintained, virus-free plants were cultured in vitro for shoot initiation on Murashige and Skoog (MS) medium containing BAP, kinetin, or 2-iP with or without IAA or NAA. Culture of shoot tips in MS with 0.57 μm IAA and node explants in MS with 2.46 μm 2-iP is recommended for `Spotti' cultivar. In `Frosty Fire', optimum number of axillary shoots was obtained from shoot tip and node explants in MS without plant regulators. Leaves and ovaries were not adequate explants for D. gratianopolitanus micropropagation because none or only a low percentage of explants regenerated shoots. High levels of cytokinins increased the number of shoots per explant but also increased the production of aberrant phenotypes and induced hyperhydricity. Adventitious shoots rooted in vitro with auxins, but maximum rooting was 97% ex vitro without auxins. This study demonstrated that D. gratianopolitanus can be successfully micropropagated. Chemical names used: 6-benzyladenine (BAP); kinetin (KIN); 6-(γ,γ-dimethylallylamino)-purine (2iP); indole-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); gibberellic acid (GA3).

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Efficient Plant Regeneration via Indirect Organogenesis in Carnation (Dianthus Caryophyllus Semperflorens flore pleno) Cultivars

Journal of Horticultural Research ◽

10.2478/johr-2021-0020 ◽

2022 ◽

Vol 0 (0) ◽

Author(s):

Hamid Reza SABAGHI ◽

Gholamreza SHARIFI-SIRCHI ◽

Pejman AZADI ◽

Mohammad Hossein AZIMI

Keyword(s):

Plant Regeneration ◽

Callus Induction ◽

Dianthus Caryophyllus ◽

Casein Hydrolysate ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Regeneration Rate ◽

Shoot Number

ABSTRACT Callus induction and plant regeneration are important steps of in vitro plant breeding of ornamental plants. In this study, the effects of different combinations of plant growth regulators (PGRs), promoters, and minerals on callus induction and plant regeneration in different carnation cultivars were studied in a completely randomized design with three replications. For callus induction, 16 different combinations of 2,4-dichlorophenoxyacetic acid (2,4-D), 6-benzylaminopurine (BA), 1-naphthaleneacetic acid (NAA), and casein hydrolysate (CH) were studied using in vitro leaf explants. The Murashige and Skoog (MS) medium supplemented with 0.2 mg·dm-3 of 2,4-D and 200 mg·dm-3 of CH showed the highest frequency of callus induction. Among the cultivars, ‘Noblesse’ showed the highest rate of callus induction (91.67%). Regarding regeneration, BA, NAA, silver nitrate (AgNO3), and adenine hemisulfate (As) were used in ten different combinations. The ‘Cameron’, ‘Tabasco’, and ‘Noblesse’ cultivars with 95.24% regeneration percentage showed the highest rate of plant regeneration. Generally, in most cultivars, the highest regeneration rate and shoot number per explant were found in the MS medium supplemented with 3 mg·dm-3 of BA, 0.6 mg·dm-3 of NAA, 5 mg·dm-3 of AgNO3, and 40 mg·dm-3 of As. According to the results, the highest regeneration frequency was obtained when 40 mg·dm-3 of As was added to the medium. Finally, the flow cytometry analysis indicated that there were no significant differences between in vitro regenerated and control plants in terms of DNA ratios.

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(280) Plant Regeneration of Periwinkle (Catharanthus roseus) from In Vitro Leaf Tissues

HortScience ◽

10.21273/hortsci.40.4.1050e ◽

2005 ◽

Vol 40 (4) ◽

pp. 1050E-1051

Author(s):

Wenhao Dai ◽

Victoria Jacques

Keyword(s):

Disease Transmission ◽

Adventitious Shoots ◽

Leaf Explants ◽

Shoot Formation ◽

Lymphocytic Leukemia ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Bedding Plant ◽

Leaf Tissues

Periwinkle, a perennial commonly used as a summer bedding plant, is known as the source of vinca alkaloids used to treat lymphocytic leukemia and Hodgkin's disease. It is also one of the natural hosts of many phytoplasma diseases, such as X-disease of major Prunus species, aster yellows, and ash yellows diseases. Therefore, periwinkle is an ideal plant species for phytoplasma disease research, such as disease transmission, species resistance, and resistant gene screening. Periwinkle tissue culture was established by incubating sterile seeds in hormone-free Murashige and Skoog (MS) medium. Plants were successfully regenerated from in vitro leaf tissues of periwinkle. Adventitious shoots were induced when leaf tissues were cultured on Murashige and Skoog (MS) medium or woody plant medium (WPM) supplemented with benzyladenine (BA) and naphthaleneacetic acid (NAA). Nearly 75% of leaf explants produced shoots in both media with 10–20 μm BA and 1 μm NAA. A mean of 4.3 shoots was produced from each explant cultured on WPM, whereas only 2 shoots were produced on MS medium under 16-h photoperiod. Leaf explants under dark treatment for 2 weeks produced big callus only, indicating that light is necessary for shoot formation. Most adventitious shoots were induced from the joint of leaf blade and petiole. In vitro shoots (>1.5 cm) were easily rooted in half-strength MS medium. Addition of NAA dramatically increased root number, with more than 20 roots being induced in 5 μm NAA medium. Rooted plants were transferred to potting medium and grown in a greenhouse.

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In Vitro Rooting and Greenhouse Acclimatization of Veltheimia bracteata and V. capensis Shoots

HortScience ◽

10.21273/hortsci.31.7.1229 ◽

1996 ◽

Vol 31 (7) ◽

pp. 1229-1230 ◽

Cited By ~ 3

Author(s):

James R. Ault

Keyword(s):

Overall Survival ◽

Potassium Salt ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Flower Bud ◽

Single Leaf ◽

Shoot Initiation ◽

Survival Percentage

Shoot initiation and multiplication were obtained in vitro from immature flower bud and leaf explants of Veltheimia bracteata Bak. `Lemon Flame' and from leaf explants of V. bracteata `Rosalba' cultured on a Murashige and Skoog (MS) medium supplemented with sucrose at 30 g•L–1, and either 8.87 μm BA plus 0.54 μm NAA or 8.87 μm BA plus 5.40 μm NAA. Shoot initiation and multiplication was obtained from a single leaf explant of Veltheimia capensis (L.) DC. on MS medium with 8.87 μm BA plus 0.54 μm NAA. Shoots of the three genotypes rooted on subculture to medium with 0.0, 4.14, or 8.29 μm K-IBA or 0.0, 4.46, or 8.92 μm K-NAA. Maximal rooting was 98% for V. bracteata `Lemon Flame', 95% for V. bracteata `Rosalba', and 98% for V. capensis, from medium with 4.46 μm KNAA. Rooted shoots were acclimatized for 3 to 4 weeks. Overall survival percentage was 69% for V. bracteata `Lemon Flame', 65% for V. bracteata `Rosalba', and 83% for V. capensis. Chemical names used: 6-benzyladenine (BA); potassium salt of indole-3-butyric acid (K-IBA); potassium salt of 1-naphthaleneacetic acid (K-NAA); 1-naphthaleneacetic acid (NAA).

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Establishment of a Rapid and Efficient Micropropagation System for Succulent Plant Haworthia turgida Haw.

HortScience ◽

10.21273/hortsci12056-17 ◽

2017 ◽

Vol 52 (9) ◽

pp. 1278-1282 ◽

Cited By ~ 4

Author(s):

Boling Liu ◽

Hongzhou Fang ◽

Chaorong Meng ◽

Ming Chen ◽

Qingdong Chai ◽

...

Keyword(s):

Callus Induction ◽

Adventitious Shoots ◽

Germplasm Conservation ◽

Leaf Explants ◽

Adventitious Shoot ◽

Induction Medium ◽

Naphthaleneacetic Acid ◽

Root Induction ◽

Ms Medium ◽

Succulent Plant

In the present study, the effect of plant growth regulators (PGRs) on callus regeneration, adventitious shoot differentiation, and root formation of Haworthia turgida Haw. was investigated. The greatest callus induction percentage (95.6%) was achieved with leaf explants inoculated on Murashige and Skoog (MS) medium with 1.0 mg·L−1 6-benzyladenine (BA) and 0.1 mg·L−1 1-naphthaleneacetic acid (NAA), and this callus induction medium supplemented with 2.5 mg·L−1 thidiazuron (TDZ) was optimal for callus proliferation. The maximum number of shoots (25.7) was obtained when the callus was cultured on MS medium supplemented with 1.0 mg·L−1 BA and 0.2 mg·L−1 2,4-dichlorophenoxyacetic acid (2,4-D). The highest number of roots per shoot (6.2) and highest rooting frequency (82.0%) were obtained when adventitious shoots were inoculated on MS medium with 0.05 mg·L−1 NAA. Regenerated plantlets were transferred to a mixture of vermiculite and soil and acclimated in a greenhouse. The survival rate of the transplanted plantlets was about 91.6%. The rate of ex vitro rooting was 83.3%, indicating that this technique is effective for root induction in H. turgida. This study has established a rapid and efficient micropropagation system that can be beneficial for commercial cultivation and germplasm conservation of H. turgida.

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

Plants ◽

10.3390/plants9060755 ◽

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.

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Induction of Callogenesis, Organogenesis, and Embryogenesis in Non-Meristematic Explants of Bleeding Heart and Evaluation of Chemical Diversity of Key Metabolites from Callus

International Journal of Molecular Sciences ◽

10.3390/ijms21165826 ◽

2020 ◽

Vol 21 (16) ◽

pp. 5826

Author(s):

Dariusz Kulus ◽

Alicja Tymoszuk

Keyword(s):

Adventitious Shoots ◽

Leaf Explants ◽

Chemical Diversity ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Explant Type ◽

Leaf Petiole ◽

Primary And Secondary Metabolites ◽

Culture Systems ◽

Lamprocapnos Spectabilis

Lamprocapnos spectabilis (L.) Fukuhara is a perennial plant species valued in the horticultural, cosmetic, and pharmaceutical markets. To date, however, there were no studies on tissue culture systems in this species when adjusted from non-meristematic explants. The aim of this study is to induce callogenesis, organogenesis, and somatic embryogenesis in non-meristematic explants of Lamprocapnos spectabilis ‘Alba’ cultured in various media and to analyze the chemical diversity of the produced callus. Leaf, petiole, and internode explants were cultured on the modified Murashige and Skoog (MS) medium fortified with various combinations and concentrations of 6-benzyladenine (BA), indole-3-acetic acid (IAA), 1-naphthaleneacetic acid (NAA), 2,4-dichlorphenoxyacetic acid (2,4-D), and picloram (PIC). After 10 weeks of culturing, the morphogenetic response of explants was evaluated and the concentration of chlorophylls, carotenoids, anthocyanins, and polyphenols in callus was analyzed. There was no influence of explant type on the callogenesis efficiency (62.1–65.3%). The highest fresh weight of callus was produced on leaf explants in the presence of 2,4-D or PIC. In contrast, the highest share of dry weight was found in internode-derived calli and cultured on IAA-supplemented medium (up to 30.8%). Only 2.5% of all explants regenerated adventitious shoots, while rhizogenesis was reported in 4.5% of explants. Somatic embryos were produced indirectly by 0% to 100% of explants, depending on the culture medium and explant type. The highest mean number of embryos (11.4 per explant) was found on petioles cultured in the MS medium with 0.5 mg·L−1 BA and 1.0 mg·L−1 PIC. Calli cultured in media with NAA usually contained a higher content of primary and secondary metabolites. There was also a significant impact of explant type on the content of anthocyanins, polyphenols, and carotenoids in callus. Further studies should focus on the elicitation of metabolites production in callus culture systems of the bleeding heart.

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REGENERATION OF TREMA ORIENTALIS (BLUME) LINN.: EFFECT OF GROWTH REGULATORS, CULTURE CONDITIONS, AND AGE AND SOURCE OF EXPLANTS

Israel Journal of Plant Sciences ◽

10.1080/07929978.1995.10676625 ◽

1995 ◽

Vol 43 (4) ◽

pp. 391-395 ◽

Cited By ~ 1

Author(s):

G.R. Rout ◽

S. Samantaray ◽

P. Das

Keyword(s):

Growth Regulators ◽

Leaf Explants ◽

Naphthaleneacetic Acid ◽

Mature Trees ◽

Regeneration Rate ◽

Bud Regeneration ◽

Shoot Bud ◽

Shoot Bud Regeneration ◽

Trema Orientalis

Optimal conditions for high frequency shoot bud regeneration from leaf callus of Trema orientalis (Blume) Linn. were studied. The regeneration rate was controlled by the growth regulators, the age and the source of the explants, and the illumination conditions. Irrespective of illumination conditions, shoot bud regeneration was achieved only in media containing benzyladenine (BA) + α-naphthaleneacetic acid (NAA) combinations, with the best results being obtained in the presence of 2.5 mg/1 BA and 0.25–0.5 mg/1 NAA. The morphogenic response was less frequent in the calluses derived from leaf explants of the mature trees compared to those of the in vitro-grown seedlings. The rate of shoot bud regeneration was more pronounced in the cultures maintained for 4 weeks in the light (16-h photoperiod) than the cultures incubated in the dark. Regenerated shoots were rooted on the medium containing 1/2 strength basal Murashige and Skoog (MS) salts supplemented with 0.01 mg/1 NAA or indole-3-butyric acid (IBA). The rooted plantlets were established in the greenhouse.

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Adventitious Shoot Regeneration and Rooting of Prunus serotina In Vitro Cultures

HortScience ◽

10.21273/hortsci.41.1.193 ◽

2006 ◽

Vol 41 (1) ◽

pp. 193-201 ◽

Cited By ~ 37

Author(s):

Ana Carolina Espinosa ◽

Paula M. Pijut ◽

Charles H. Michler

Keyword(s):

Shoot Regeneration ◽

Adventitious Shoots ◽

Leaf Explants ◽

Adventitious Shoot ◽

Prunus Serotina ◽

Naphthaleneacetic Acid ◽

Eastern United States ◽

Continuous Darkness ◽

Number Of Shoots

A complete regeneration protocol was developed for Prunus serotina Ehrh., an important hardwood species for timber and sawlog production in the central and eastern United States. Nodal sections were cultured on Murashige and Skoog (MS) medium supplemented with 4.44 μm 6-benzylaminopurine (BA), 0.49 μm indole-3-butyric acid (IBA), and 0.29 μm gibberellic acid (GA3). In vitro leaf explants of three genotypes were placed on woody plant medium (WPM) supplemented with 0, 2.27, 4.54, or 6.81 μm thidiazuron (TDZ) in combination with 0, 0.54, 1.07, or 5.37 μm naphthaleneacetic acid (NAA), and on WPM supplemented with 0, 4.44, 8.88, or 13.32 μm BA in combination with 0, 0.54, 1.07, or 5.37 μm NAA. Cultures were maintained either in continuous darkness for 5 weeks, or in the dark for 3 weeks and then transferred to a 16-hour photoperiod. TDZ and the genotype had a significant effect on the number of shoots regenerated. The maximum mean number of shoots regenerated per explant (5.05 ± 1.14) was obtained with 2.27 μm TDZ plus 0.54 μm NAA with the 3-week dark period then light treatment. The highest percent shoot regeneration (38.3) and mean number of shoots (4.13 ± 0.97) was obtained with 6.81 μm TDZ plus 1.07 μm NAA. The highest rooting (27%) of adventitious shoots and number of roots per shoot (2.3 ± 0.2) was obtained with 2.5 μm IBA when shoots were maintained for 7 days in the dark on rooting medium before transfer to a 16-hour photoperiod. The highest rooting (70%) of nodal explant-derived stock cultures and number of roots per shoot (2.7 ± 0.9) was also obtained with 2.5 μm IBA, but when shoots were maintained for 4 days in the dark before transfer to a 16-hour photoperiod. In total, 86% of the plantlets survived acclimatization to the greenhouse and 100% survival after overwintering in cold-storage.

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