scholarly journals High-frequency Direct Somatic Embryogenesis and Plantlet Regeneration from Leaves Derived from In Vitro-Germinated Seedlings of a Coffea arabica Hybrid Cultivar

HortScience ◽  
2016 ◽  
Vol 51 (9) ◽  
pp. 1148-1152 ◽  
Author(s):  
Jane Kahia ◽  
Margaret Kirika ◽  
Hudson Lubabali ◽  
Sinclair Mantell
Keyword(s):  
Tissue Culture ◽  
High Frequency ◽  
Somatic Embryos ◽  
Coffea Arabica ◽  
Alternative Methods ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Embryogenic Cultures ◽  
Half Strength

Breeding work carried out during the period 1971–85 by the Coffee Research Institute, Ruiru, Kenya resulted in the release of a new improved hybrid Coffea arabica named Ruiru 11. The cultivar combines resistance to coffee berry disease (CBD) and leaf rust, with high yield and good cup quality attributes. The propagation by F1 hybrid seeds production, cuttings, and tip grafting do not produce enough planting materials. There was a need to explore alternative methods and tissue culture offers potential options. The objective of the study was to evaluate the effect of explant sources and cytokinins on induction and regeneration of somatic embryos. Eight different explants were cultured on half-strength Murashige and Skoog (MS) medium supplemented with 10 µm benzylaminopurine (BAP). The effect of kinetin, N6-(2-isopentyl) adenine (2iP) evaluated at (0, 0.5, 5, or 25 µm) or thidiazuron (TDZ) (0, 0.5, 1.0, or 5 µm) added in separate experiments was also evaluated. The percentage of embryogenic cultures and the numbers of embryos per explant were determined after 3 months’ culture. The explant type had a significant effect (P > 0.05) on the induction of somatic embryos. Explants from in vitro-germinated seedlings produced the highest embryogenic cultures (90%) and the highest mean number of embryos (19.36) per explant. Cytokinins strongly enhanced induction and regeneration of somatic embryos. TDZ at 1 µm produced the highest embryogenic cultures (100%) and the highest mean number of embryos (24.2). The embryos were germinated on half-strength MS medium without any hormones. A high (98%) survival rate of the regenerated plantlets was recorded over all the treatments in the greenhouse. This is the first report on induction of high-frequency direct somatic embryos from coffee juvenile tissues. This is of great significance in tissue culture and indeed molecular biology manipulations because it allows regeneration of coffee from several explants.

scholarly journals Micropropagation, Micromorphological Studies, andIn VitroFlowering inRungia pectinataL.

Scientifica ◽  
2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari ◽  
C. P. Ravindran
Keyword(s):  
Tissue Culture ◽  
Shoot Multiplication ◽  
Developmental Changes ◽  
Bud Break ◽  
Ms Medium ◽  
Half Strength ◽  
Indole 3 Acetic Acid ◽  
Culture Protocol ◽  
Important Medicinal Plant

A tissue culture protocol was developed for an important medicinal plantRungia pectinataL. in the present study. Nodal shoots were used as explants and surface-sterilized with 0.1% HgCl2solution. Murashige and Skoog (MS) medium was used to establish the cultures ofR. pectinata. The bud break was reported on MS medium supplemented with 1.0 mg L−16-benzylaminopurine (BAP). About 98% response was observed with this media combination and maximum 3.2 shoots per explant with 4.3 cm length were recorded. The shoots were further multiplied using MS medium augmented with 0.5 mg L−1each of BAP and kinetin (Kin) + 0.1 mg L−1indole-3 acetic acid (IAA). Maximum 13.2 shoots per explant with 5.2 cm length were observed. All the shoots were rooted (4.9 roots per shoot with 3.5 cm length) on half strength MS medium fortified with 2.0 mg L−1indole-3 butyric acid (IBA).In vitroflowering was induced from the shoots on half strength MS medium supplemented with same concentrations and combinations of growth regulators used for shoot multiplication under 12/12 hr light/dark photoperiod. The plantlets were hardened in the greenhouse for two months and finally transferred to the field. The foliar micromorphological studies revealed the developmental changes in stomata, vein density, and trichomes during the culture of shoots underin vitroconditions.

scholarly journals Effect of Plant Growth Regulators on Somatic Embryogenesis and Plantlet Development of Turkey Berry (Solanum torvum SW)

2021 ◽  
pp. 1-8
Author(s):  
Ghan Singh Maloth ◽  
Rajinikanth Marka ◽  
Rama Swamy Nanna
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Solanum Torvum ◽  
Plantlet Formation ◽  
Regenerated Plants

In the present study it was reported on direct somatic embryogenesis and plant regeneration from cotyledon and leaf explants of Turkey berry/pea egg plant (Solanum torvum SW), a medicinally important plant. Somatic embryogenesis has several advantages over other routes of in vitro plant regeneration. Somatic embryogenesis was induced directly from cotyledon and leaf explants on MS medium fortified with BAP (0.5 mg/L)+NAA (0.5-6.0 mg/L). High percentage of somatic embryogenesis (90%), maximum number of somatic embryos formation (62±0.18)  along with high percentage (76%) conversion of somatic embryos into bipolar embryos was observed on cotyledon explants in 0.5 mg/L BAP+2.5 mg/L NAA. At the same concentration of BAP (0.5 mg/L)+NAA (2.5 mg/L) also resulted  on the maximum percentage of somatic embryogenesis (92%), the highest number of somatic embryos formation (88±0.15) and the highest percentage (76%) of somatic embryos conversion into bipolar embryos in leaf explants. A mixture of globular, heart and torpedo-shaped embryos were germinated on MS medium supplemented with 0.5 mg/L IAA+1.0-4.0 mg/L BAP. Maximum germination frequency (75±0.14) of somatic embryos and plantlet formation was found in 0.5 mg/L IAA+2.0 mg/L BAP, but they didn’t germinate on ½ MSO and MSO media. The survival rate of regenerated plants after field transfer was recorded to be 75%. These regenerated plants were found morphologically similar to donor plants. The present protocol can be used for conservation of the species and also for genetic transformation experiments in S. torvum.

scholarly journals In vitro Micropropagation of Pomegranate (Punica granatum L.) Derived from Cotyledon

2021 ◽  
Vol 31 (1) ◽  
pp. 61-69
Author(s):  
MH Kabir ◽  
Pronabananda Das ◽  
ANK Mamun ◽  
Md Monirul Islam ◽  
Md Aminul Islam
Keyword(s):  
High Frequency ◽  
Punica Granatum ◽  
Ms Medium ◽  
In Vitro Plant Regeneration ◽  
Culture Initiation ◽  
In Vitro Micropropagation ◽  
Half Strength ◽  
Acclimatization Period ◽  
Punica Granatum L

A high frequency in vitro plant regeneration of pomegranate was established on MS medium supplemented with different concentrations and combinations of plant growth regulators. As explant cotyledons were employed for this study. Ninety percent of the cultured explants responded to form shoots from 30 days old in vitro raised seedlings after 90 days of culture initiation in MS containing 1.0 mg/l IBA + 0.1 mg/l NAA. The average number of shoots per explant was 10.0 ± 2.20, shoot length of 12.0 ± 2.40 cm, node per regenerated shoot was 9.0 ± 1.60 and the leaf number was14.0 ± 1.40. Well developed shoots were cultured on half strength of MS medium supplemented with 0.5 mg/l IBA, in which 90% shoot induced roots implanted after one month. The average number of root per shoot was 8.0 ± 0.90 and the average root length of 6.5 ± 0.40 cm was observed in this medium. Eighty percent plantlets were survived in the outdoor condition during the acclimatization period of seven days. Plant Tissue Cult. & Biotech. 31(1): 61-69, 2021 (June)

scholarly journals Can Agricultural Biotechnology Help Guava Growing in Temperate Climate

HortScience ◽  
2004 ◽  
Vol 39 (4) ◽  
pp. 861B-861
Author(s):  
Bipul K. Biswas* ◽  
Nirmal Joshee ◽  
Anand K. Yadav
Keyword(s):  
Tissue Culture ◽  
Somatic Embryos ◽  
Adventitious Shoots ◽  
Multiple Shoots ◽  
Cold Climate ◽  
Nodal Explants ◽  
Temperate Climate ◽  
Zygotic Embryos ◽  
Ms Medium

Guava (Psidium guajava L.), also called `apple of tropics,' is immensely nutraceutical and horticulturally important. Being a tropical plant, it cannot stand temperatures below 25° F and needs frost protection to grow in temperate regions. To adapt in cold climate, cold hardy guava cultivars are needed. Conventional ways are uneconomic in time and efforts. Still, transgenic plants developed using biotechnological approaches of tissue culture and rDNA technology, appear to have great potential. Thus, protocols for in vitro propagation of guava were developed via organogenesis and somatic embryogenesis using nodal explants from mature trees and young zygotic embryos, respectively. Nodal explants induced multiple shoots when cultured on MS medium fortified with KIN, BAP and Ad.S. Adding a (NO3)2 to medium was useful to prevent in vitro shoot tip browning of adventitious shoots. Rocker liquid culture greatly increased growth of multiple shoots compared to the agar-based medium. It appears to be a good tool for woody plant tissue culture. Induction of somatic embryos in guava was also achieved on MS medium supplemented with IAA auxin. About 80% to 90% somatic embryos germinated normally. To achieve Agro-bacterium-mediated gene transfer in guava, on-going co-cultivation of organogenic tissues of guava is to optimize protocols for freeze tolerance gene (CBF1, CBF2, CBF3) transfer. Plasmid vectors containing selectable markers (nptII gene for antibiotic selection and GUS reporter gene as scorable gene mediated selection), with CaMV 35S promoter gene has been introduced into guava tissues and the resultant plants showed antibiotic resistance. Details of the experimental procedures and up-to-date results will be discussed.

scholarly journals (291) Plant Regeneration through Direct Somatic Embryogenesis in Homalomena `Emerald Gem'

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1078A-1078
Author(s):  
Qian Zhang ◽  
Jianjun Chen ◽  
Richard J. Henny
Keyword(s):  
Somatic Embryogenesis ◽  
Plant Regeneration ◽  
Somatic Embryo ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Multiple Shoot ◽  
Direct Somatic Embryogenesis ◽  
Induction Medium ◽  
Ms Medium

Homalomena `Emerald Gem' is an important ornamental foliage plant and widely used for interior plantscaping. Current propagation of this cultivar has been primarily carried out through in vitro culture by organogenesis; regeneration through somatic embryogenesis has not been documented. This report describes successful plant regeneration via direct somatic embryogenesis from explants of different organs. Somatic embryos formed at and around the cut surface of petiole, spathe, and peduncle explants. Embryos also appeared at the base between expanded ovaries of the spadix segment, and around midrib of leaf explants. The optimal treatments for somatic embryo occurrence from petiole, spathe, and peduncle explants were MS medium containing 0.2 mg/L NAA or 0.5 mg/L 2, 4-D with 2.0 mg/L CPPU, and for spadix explants were MS medium with 0.5 mg/L PAA and 2.5 mg/L TDZ. Somatic embryos appeared 6 to 8 weeks after culture and formed large embryo clumps in 3 to 4 months. Somatic embryos produced more secondary embryos and geminated on induction medium. Multiple shoot development and plant regeneration occurred from somatic embryo clusters on MS medium without hormone or with 2 mg/L BA and 0.2 mg/L NAA. The regenerated plants grew vigorously after transplanting to a soilless container substrate in a shaded greenhouse.

scholarly journals In Vitro Regeneration and Mass Propagation of Ruta graveolens L.—A Multipurpose Shrub

HortScience ◽  
2005 ◽  
Vol 40 (5) ◽  
pp. 1478-1480 ◽  
Author(s):  
Mohd Faisal ◽  
Naseem Ahmad ◽  
Mohammad Anis
Keyword(s):  
High Frequency ◽  
In Vitro Regeneration ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ruta Graveolens ◽  
Mass Propagation ◽  
Regeneration Frequency ◽  
Whole Plant ◽  
Half Strength

A protocol for rapid in vitro propagation of Ruta graveolens L. through high-frequency shoot induction from nodal explants was established. Proliferation of shoots from nodal segments was achieved on Murashige and Skoog medium supplemented with various concentrations of BA, Kin, IAA, and NAA, either singly or in various combinations. The highest shoot regeneration frequency (98.5%) and the highest number of shoots per explant (40.2 ± 2.8) was obtained on MS medium supplemented with 10 μm BA and 2.5 μm NAA. In vitro regenerated shoots rooted best on half-strength MS medium containing 0.5 μm IBA. Rooted shoots, following acclimatization in the greenhouse, were successfully transferred to field conditions, and 90% of plants survived. The efficient in vitro regeneration of the whole plant can be used as a fast and reliable method to transform R. graveolens genetically for its active principles.

scholarly journals (80) Direct Somatic Embryogenesis from Thin-section Leaf Explant of Phalaenopsis Hybrids

HortScience ◽  
2006 ◽  
Vol 41 (4) ◽  
pp. 1066D-1067
Author(s):  
Jae-Dong Chung ◽  
Hong-Yul Kim ◽  
Jung-Hae Suh ◽  
Oh-Chang Kwon ◽  
Chang-kil Kim
Keyword(s):  
Somatic Embryogenesis ◽  
Somatic Embryo ◽  
Thin Section ◽  
Somatic Embryos ◽  
Leaf Explants ◽  
Leaf Explant ◽  
Direct Somatic Embryogenesis ◽  
Ms Medium ◽  
Embryo Formation ◽  
Half Strength

Somatic embryo formation was observed on thin-sectioned leaf explants within 3 weeks of culture from two Phalaenopsis hybrids—Phalaenopsis Hwafeng Redjewell `Ching Ruey' Phalaenopsis Chingruey's Giant Ching Ruey' (R×R), and Phalaenopsis Formosa Best Girl Ching Ruey' Depts. Lih Jiang Beauty `S 566' (WR×WR). Frequency of somatic embryo formation was higher in hybrid WRxWR than R×R and optimal concentration of TDZ for the induction of somatic embryos was 9.08 μM. In (WR×WR) embryo proliferation was simultaneously observed after transferring the explants with somatic embryo clumps onto PGR-free half-strength MS medium. Six months after initiation, the culture plantlets were produced. This is the first report on somatic embryogenesis induced directly from the leaf explants using TDZ in Phalaenopsis.

scholarly journals In Vitro Flowering of Plantlets Regenerated from Zygotic Embryo-derived Somatic Embryos of Ginseng

HortScience ◽  
1990 ◽  
Vol 25 (12) ◽  
pp. 1652-1654 ◽  
Author(s):  
Haeng S. Lee ◽  
Jang R. Liu ◽  
Seung G. Yang ◽  
Young H. Lee ◽  
Kwang-W. Lee
Keyword(s):  
Somatic Embryos ◽  
Zygotic Embryo ◽  
Ginseng Root ◽  
Zygotic Embryos ◽  
Ms Medium ◽  
Root Explants ◽  
Half Strength ◽  
Dichlorophenoxy Acetic Acid ◽  
Somatic Embryo Induction

Mature zygotic embryos dissected from ginseng (Panax ginseng C.A. Meyer) seeds were cultured on Murashige and Skoog (MS) medium containing various concentrations of 2,4-D and kinetin. Somatic embryos were induced directly from cotyledonary tissue and from intervening callus. The frequency of somatic embryo induction was up to 55% of zygotic embryo explants. Upon transfer onto half-strength MS medium supplemented with 1 mg BA/liter and 1 mg GA3/liter, most somatic embryos developed into plantlets. More than 50% of the plantlets flowered after 4 weeks of culture, and some developed immature fruits in vitro. These results indicate that adulthood of ginseng root explants is not a prerequisite for flowering of plantlets regenerated through somatic embryogenesis. Chemical names used: (2,4 -dichlorophenoxy) acetic acid (2,4-D); N-(2-furanylmethyl) -1H-purin-6-amine(kinetin); N-(phenylmethyl) -1H-purin-6-amine (BA); gibberellic acid (GA3).

scholarly journals In vitro multiplication, micromorphological studies and ex vitro rooting of Hybanthus enneaspermus (L.) F. Muell. – a rare medicinal plant

2018 ◽  
Vol 77 (1) ◽  
pp. 80-87 ◽  
Author(s):  
Mahipal S. Shekhawat ◽  
M. Manokari
Keyword(s):  
Medicinal Plant ◽  
Large Scale ◽  
Nodal Segments ◽  
Ms Medium ◽  
Ex Vitro ◽  
Culture Bottle ◽  
Hybanthus Enneaspermus ◽  
Half Strength

AbstractHybanthus enneaspermusis a rare medicinal plant. We defined a protocol for micropropagation,ex vitrorooting of cloned shoots and their acclimatization. Surface-sterilized nodal segments were cultured on Murashige and Skoog (MS) medium with different concentrations of 6-benzylaminopurine (BAP) and kinetin (Kin). Medium supplemented with 1.5 mg L−1BAP was found optimum for shoot induction from the explants and 6.4±0.69 shoots were regenerated from each node with 97% response. Shoots were further proliferated maximally (228±10.3 shoots per culture bottle with 7.5±0.43 cm length) on MS medium augmented with 1.0 mg L−1each of BAP and Kin within 4–5 weeks. The shoots were rootedin vitroon half strength MS medium containing 2.0 mg L−1indole-3 butyric acid (IBA). The cloned shoots were pulse-treated with 300 mg L–1 of IBA and cultured on soilrite® in a greenhouse. About 96% of the IBA-pulsed shoots rootedex vitroin soilrite®, each shoot producing 12.5±0.54 roots with 5.1±0.62 cm length. Theex vitrorooted plantlets showed a better rate of survival (92%) in a field study thanin vitrorooted plantlets (86%). A comparative foliar micromorphological study ofH. enneaspermuswas conducted to understand the micromorphological changes during plant developmental processes fromin vitrotoin vivoconditions in terms of variations in stomata, vein structures and spacing, and trichomes. This is the first report onex vitrorooting inH. enneaspermusand the protocol can be exploited for conservation and large-scale propagation of this rare and medicinally important plant.

scholarly journals An Efficient Protocol for High-frequency Direct Multiple Shoot Regeneration from Internodes of Peppermint (Mentha x piperita)

2010 ◽  
Vol 5 (12) ◽  
pp. 1934578X1000501
Author(s):  
Sanjog T. Thul ◽  
Arun K. Kukreja
Keyword(s):  
Shoot Regeneration ◽  
Multiple Shoots ◽  
Shoot Elongation ◽  
Shoot Formation ◽  
Multiple Shoot ◽  
Ms Medium ◽  
Mentha X Piperita ◽  
Half Strength ◽  
Multiple Shoot Regeneration

A simple, repeatable and efficient protocol for direct multiple shoot regeneration from internodal explants has been defined in peppermint ( Mentha x piperita var. Indus). In vitro regenerated shoots of peppermint were excised into 4 to 8 mm long internodes and cultured on Murashige and Skoog's medium supplemented with different cytokinins. In the hormonal assay, 3.0 mg L-l zeatin or 6-isopentenyl adenine independently supplemented to half strength MS medium exhibited multiple shoot regeneration, while thiaduzorn (0.1-3.0 mg L−1) showed no morphogenetic effect. A maximum of 85% in vitro cultured explants showed multiple shoot formation with an average of 7 shoots per explant on MS medium supplemented with zeatin. Multiple shoots were initiated within three weeks of cultivation. Internodes with regenerated multiple shoots were transferred to half - strength MS medium without supplementing with any plant growth hormone for shoot elongation and rhizogenesis. Rooted plants acclimatized and grew to maturity under glasshouse conditions. The plantlets developed were phenotypically identical to the parent plant and exhibited 96 % survival.

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