In vitro propagation and growth of ex vitro Chrysanthemum indicum L. in Da Lat - Lam Dong

Chrysanthemum indicum plant is a valuable and beneficial herb for human health. In this paper, we present the results of in vitro propagation of this plant in Da Lat. The results showed that MS medium supplemented with 25 g/l sucrose, 9 g/l agar, pH 5.8 was the best for shoot regeneration and growth (shoot height was 2.41 - 2.47 cm, 1 shoot/explant). MS media supplemented with BA (0.1, 0.5, 1, 1.5, 2 mg/l), Kinetin (0.1, 0.5, 1, 1.5, 2 mg/l ) and TDZ (0.1, 0.5, 1 mg/l) were unsuitable for shoot regeneration and growth of C. indicum. The regeneration and growth of shoots on the medium supplemented with 1 g/l of activated charcoal was better than (plant height of 3.45 cm) medium without activated charcoal (plant height of 2.46 cm). MS medium supplemented with 0, 0.1, 0.5, 1 mg/l IBA, 25 g/l sucrose, 9 g/l agar, pH 5,8 were suitable for in vitro root regeneration, rate of root formation was 100%. Coconut fiber powder was the best substrate to transfer the plantlets to the greenhouse, with survial rate of 100%, plantlets grew well. Nitrophoska irrigation was the best for the growth and development of C. indicum cultivated on coconut fiber powder under greenhouse conditions (plant height of 61.70 cm, 100.80 flowers/tree, flower diameter of 1.68 cm, fresh weight of 0.376 g/flower). The results also show that seedlings derived from tissue culture can be used to cultivate C. indicum and C. indicum grew well in the climate of Da Lat - Lam Dong and flowered all year round.

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Study on in vitro propagation of Sophora tonkinensis Gagnep through stem node culture

Ministry of Science and Technology, Vietnam ◽

10.31276/vjst.63(12).59-63 ◽

2021 ◽

Vol 63 (12) ◽

pp. 59-63

Author(s):

Thi Lai Nguyen ◽

◽

Thi Binh Nguyen ◽

Anh Duc Pham ◽

Huong Son Pham ◽

...

Keyword(s):

Activated Charcoal ◽

Coconut Water ◽

High Survival ◽

Ms Medium ◽

Shoot Height ◽

Sophora Tonkinensis ◽

Carrot Puree ◽

Node Culture ◽

Stem Segments

In the present study, authors propagated Sophora tonkinensisGagnep plants using stem nodal culture. The results indicated that on MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 0.75 mg/l TDZ shoots proliferated from stem segments have the best count and height of shoots. The most appropriate medium for multiplication of shoots was the MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 0.75 mg/l TDZ, 0.5 mg/l IBA, 2.0 g/l peptone, 30 g/l carrot puree, pH 5.5 with the results of 20.60 shoots/explant, shoot height of 3.75 cm and 4.6 leaves/shoot after 8 weeks of culture. Root formation of shoots carried out on the MS medium supplemented with 30 g/l sucrose, 5.5 g/l agar, 200 ml/l coconut water, 1 g/l activated charcoal, 1.0 mg/l αNAA gave the best result. In the nursery, a mixture of humus + coconut fiber powder (70:30 ratio) was regarded as the best substrate due to the high survival rate of plantlets (92%) and healthy plantlets (10.30 cm high with 7.2 leaves and 4.3 new roots/a plantlet) at 10 weeks after planting

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Explants selection for in vitro propagation of Pachyrhizus erosus L.

Notulae Scientia Biologicae ◽

10.15835/nsb13110844 ◽

2021 ◽

Vol 13 (1) ◽

pp. 10844

Author(s):

Idowu A. OBISESAN ◽

Ayobola M. A. SAKPERE ◽

Bamidele J. AMUJOYEGBE ◽

Michael S. AKINROPO

Keyword(s):

Shoot Regeneration ◽

In Vitro Propagation ◽

Callus Formation ◽

Leaf Explants ◽

Stem Explants ◽

Ms Medium ◽

Friable Callus ◽

Pachyrhizus Erosus ◽

Regeneration Response

Pachyrhizus erosus tuber is rich in protein asides its agronomical value as a legume, but the seeds by which it is propagated have very low viability. This study established sterilization protocol and effect of various concentrations of auxins and cytokinins on callus production and shoot regeneration from explants of P. erosus. Explants and seeds were sterilized using sodiumhypochlorite (NaClO) solution (5, 10 and 15% v/v) for 5 and 10 mins. Nodal, stem and leaf explants from in vitro germinated P. erosus and tuber from field grown plant were sterilized and cultured on Murashige and Skoog (MS) medium (control) and MS combined with different concentrations of auxins (NAA and 2, 4-D) and cytokinin (BA and Kinetin) and the cultured explants were monitored in terms of degree of callus formation, morphology and colour of callus and also for shoot induction. The results showed that seeds of P. erosus sterilized with 10% NaClO solution for 10 mins and germinated in vitro is the best way of getting sterile nodal, stem and leaf explants for the in vitro propagation of the plant, while tuber explants could be sterilized with 15% NaClO for 10 minutes. Nodal explants inoculated in MS medium supplemented with 1.0 mg/L BA gave the highest shoot regeneration response, while stem explants inoculated on MS medium supplemented with 1.0 mg/L BA and 0.5 mg/L NAA also gave the highest amount of friable callus. The study concluded that in vitro germinated seeds were the best way of getting explant for P. erosus.

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Indirect shoot regeneration of Iranian purple coneflower (Echinacea purpurea L.) from cotyledon and hypocotyl explants

Acta Agronomica Hungarica ◽

10.1556/aagr.59.2011.1.7 ◽

2011 ◽

Vol 59 (1) ◽

pp. 65-72 ◽

Cited By ~ 2

Author(s):

A. Zebarjadi ◽

J. Motamedi ◽

A. Ismaili

Keyword(s):

Shoot Regeneration ◽

Callus Induction ◽

Shoot Formation ◽

Ms Medium ◽

Medicinal Species ◽

Regeneration Rate ◽

Hypocotyl Explants ◽

Purple Coneflower ◽

Significant Difference

In vitro plant regeneration was optimized for Iranian purple coneflower via organogenesis from callus cultures derived from cotyledon and hypocotyl tissues by placing them on MS medium supplemented with different concentrations and combinations of BAP and NAA. The experiment was laid out as a completely randomized design in a factorial arrangement with three replications. The results indicated that the mean callus induction was influenced by explant type, with a significant difference between cotyledon (77.81%) and hypocotyl (65.33%) explants at the 0.01 probability level. In relation with the regeneration rate, no significant differences were observed between the two types of explants. For both cotyledons and hypocotyls the optimum shoot regeneration frequency (31.5% and 32.5%, respectively) and number of shoots per explant (5.2 and 5.3, respectively) were achieved using medium supplemented with 0.4 mg l−1 BAP. Proliferated shoots were elongated in hormone-free MS medium and well-developed shoots were rooted on MS medium, both with and without the addition of 2 mg l−1 IBA. All the plantlets survived acclimatization, producing normal plants under controlled conditions. This study revealed that cotyledon and hypocotyl explants of E. purpurea have relatively good potential for callus induction and shoot formation. Furthermore, a beneficial method has been established for the micropropagation of this valuable medicinal species.

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Nutritional requirements for germination and in vitro development of three Orchidaceceae species in the southern Brazilian Amazon

Ornamental Horticulture ◽

10.14295/oh.v24i2.1130 ◽

2018 ◽

Vol 24 (2) ◽

pp. 87-94 ◽

Cited By ~ 1

Author(s):

Viviane Luiza Hunhoff ◽

Lais Alves Lage ◽

Ednamar Gabriela Palú ◽

Willian Krause ◽

Celice Alexandre Silva

Keyword(s):

Activated Charcoal ◽

Culture Media ◽

Leaf Number ◽

Alternative Form ◽

Ms Medium ◽

Root Number ◽

Shoot Height ◽

Full Strength ◽

Half Strength

Tissue culture is an alternative form of producing healthy, vigorous and regular plants on a large scale. The purpose of this study was to evaluate the most efficient culture medium for in vitro plantlet germination and development of three Orchidaceae species. Seeds disinfested of three species were dispersed in distilled water and dripped into basic Murashige and Skoog (MS) medium. The experimental design was completely randomized in a factorial 3 x 4 (three species x four culture media), with 5 replications. Four treatments were established: (1) full-strength MS medium, with the full nutrient concentration (MSØ), (2) full-strength MS medium plus 0.3% activated charcoal (MSØ ACh), (3) half- strength MS medium (½ MS) and (4) half- strength MS medium with 0.3 % activated charcoal (½ MS ACh). Germination was evaluated after 15, 20, 25, 30, and 60 days. The shoot height, leaf number and length, root number and length of plantlets of the three studied species were assessed. In A. variegata, 73% germinated after 60 days in ½ MS ACh medium. In the same period, 100% of E. viparum and S. gloriosa seeds germinated in MSØ ACh medium. The plant height, leaf number and length, root number and length were significantly higher for the species A. variegata and E. viviparum in MSØ ACh medium. The culture media ½ MS and MSØ with addition of activated charcoal favored in vitro germination for the three orchid species of this study.

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In vitro propagation of the Chinese traditional and medicinal plant Heracleum scabridum Franch

Bangladesh Journal of Botany ◽

10.3329/bjb.v48i3.47933 ◽

2019 ◽

Vol 48 (3) ◽

pp. 575-581

Author(s):

Li-Juan Zou ◽

Jin-Yao Hu ◽

Ming-Hua Luo ◽

Qing-Gui Wu

Keyword(s):

Medicinal Plant ◽

Shoot Regeneration ◽

In Vitro Propagation ◽

Multiple Shoots ◽

Axillary Bud ◽

Direct Organogenesis ◽

Ms Medium ◽

Indirect Organogenesis ◽

Best Response

Three explants such as stem tips, leaves and petioles of Heracleum scabridum were compared for their shoot development/differentiation ability by using different plant growth regulators (PGRs). The most effective PGRs combination for callus induction and organogenesis was determined. TDZ, Kn, Zn and IAA were used to induce multiple shoots. For indirect organogenesis (from the calli), the best response (27.25 ± 4.19 shoot per stem tips) and (18.23 ± 2.12 per leaves) were obtained in Murashige and Skoog (MS) medium fortified with 0.5 mg/l IAA and 3.0 mg/l Zn with 100, 97.3% induction rate, respectively. MS medium containing 0.5 mg/l IAA and 3.0 mg/l Zn was also found to be optimal for shoot regeneration from petioles. The highest percentage of regeneration (94.6) with mean number of shoots 17.83 ± 4.87 from petioles were produced. For direct organogenesis (from axillary bud shoot clumps), 0.1 mg/l IAA and 1.5 mg/l TDZ were found to be optimal for shoot regeneration of stem tips, with mean numbers of axillary bud shoot clumps 7.12 ± 1.23 were produced. Plantlets were transferred to soil with 95% of plants acclimatized recovered successfully.

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Protokol Perbanyakan Masal Dendrobium ‘Balithi CF22-58’ secara In Vitro Melalui Embriogenesis Somatik Tidak Langsung (In Vitro Propagation Protocol of Dendrobium ‘Balithi CF22-58’ via Indirect Somatic Embryogenesis)

Jurnal Hortikultura ◽

10.21082/jhort.v29n2.2019.p137-146 ◽

2020 ◽

Vol 29 (2) ◽

pp. 137

Author(s):

Fitri Rachmawati ◽

Dewi Permanik ◽

Ronald Bunga Mayang ◽

Budi Winarto

Keyword(s):

Somatic Embryogenesis ◽

Activated Charcoal ◽

Embryogenic Callus ◽

Culture System ◽

In Vitro Propagation ◽

Clonal Propagation ◽

Ms Medium ◽

Indirect Somatic Embryogenesis ◽

Half Strength

Protokol perbanyakan klonal yang efektif dan efisien sangat diperlukan untuk produksi benih berkualitas pada komersialisasi produk unggulan hasil pemuliaan. Penelitian bertujuan untuk mendapatkan protokol perbanyakan klonal Dendrobium ‘Balithi CF22-58’ melalui embriogenesis tidak langsung. Percobaan dilakukan di Laboratorium Kultur Jaringan Balai Penelitian Tanaman Hias dari bulan Januari hingga Desember 2017. Penelitian ini menekankan pada penggunaan jenis eksplan, media, dan sistem kultur. Jenis eksplan yang diuji adalah tunas pucuk, tunas lateral, dan pangkal plantlet dengan tiga media inisiasi [½ Murashige and Skoog (MS) dikombinasikan dengan 1,5 mg/l thidiazuron (TDZ) dan 0,5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2,5 mg/l metathopolin (mT) dan 0,05 mg/l BAP (MI-2), dan 5 mg/l mT dan 0,05 mg/l BAP (MI-3)]; empat media proliferasi, yaitu ½ MS dengan kombinasi: MP-1 (0,75 mg/l TDZ + 0,25 mg/l BAP), MP-2 (1,5 mg/l TDZ + 0,5 mg/l BAP), MP-3 (2,5 mg/l mT + 0,05 mg/l BAP), dan MP-4 (5,0 mg/l+ 0,05 mg/l BAP); dua sistem kultur (padat dan cair); dan tiga media regenerasi MPP-1 (½ MS dengan vitamin penuh (1/2 MS-FV) + 2% charcoal); MPP-2 (½ MS-FV); dan MPP-3 (2 g/l Rosasol 18:18:18 TE). Percobaan disusun menggunakan rancangan acak kelompok faktorial dengan lima ulangan. Hasil penelitian menunjukkan bahwa inisiasi kalus embriogenik (KE) tertinggi, yaitu 38,3% dengan waktu inisiasi 16,8 hari dihasilkan dari eksplan pangkal plantlet pada medium MI-1. Medium MP-2 dan sistem kultur cair mampu mempertahankan proliferasi KE sampai 83,1% dengan rasio penggandaan 3,23 kali. Perkecambahan embrio terbaik sampai 86,9% embrio berkecambah dengan 18,2 kecambah per rumpun dalam waktu 21,3 hari, ditunjukkan pada medium MPP-1, sedangkan pembesaran plantlet terbaik mencapai tinggi plantlet sampai 5 cm, jumlah daun hingga 4,9 helai, dan jumlah akar 2,8, dengan 2,6 cm panjang akar dan 0,27 g bobot basah plantlet, diperoleh pada medium MPP-3. Perbanyakan anggrek dengan protokol ini diperkirakan dapat menghasilkan sekitar 3.000–4.000 plantlet/eksplan/tahun. Protokol hasil penelitian ini sangat potensial diaplikasikan pada perbanyakan klonal Dendrobium melalui kultur jaringan. KeywordsDendrobium; Embriogenesis somatik; Perbanyakan masal; Proliferasi; Sistem kultur AbstractThe effective and efficient clonal propagation protocol is significantly needed for producing qualified seedling for commercialization of superior breeding products. The objective of the study was to establish clonal propagation protocol for Dendrobium ‘Balithi CF22-58’ via indirect somatic embryogenesis. The study was conducted at the Tissue Culture Laboratory in Indonesian Ornamental Crops Research Institute from January to December 2017. The study emphasized to utilize explant source, culture media, and culture system. Explant types were shoot tip, lateral shoot, and basal part of plantlets; three initiation media [half strength Murashige and Skoog (MS) medium containing 1.5 mg/l thidiazuron (TDZ) and 0.5 mg/l 6-benzylaminopurine (BAP) (MI-1), 2.5 mg/l metathopolin (mT) and 0.05 mg/l BAP (MI-2), and 5 mg/l mT and 0.05 mg/l BAP (MI-3)]; four proliferation media (half strength MS medium supplemented with: MP-1 (0.75 mg/l TDZ and 0.25 mg/l BAP), MP-2 (1.5 mg/l TDZ and 0.5 mg/l BAP), MP-3 (2.5 mg/l mT and 0,05 mg/l BAP), and MP-4 (5.0 mg/l and 0.05 mg/l BAP); two culture system were solid and liquid; and three regeneration media viz, MPP-1 (half strength MS medium with full vitamin and 2% activated charcoal); MPP-2 (MR-1 activated charcoal free), and MPP-3 (2 g/l Rosasol 18:18:18 TE). These experiments were arranged using a factorial randomized complete block design with five replications. Results of the study revealed that the highest initiation rate of embryogenic callus (EC) was up to 38.3% in 16.8 days after culture. The EC was regenerated from a basal part on MI-1 medium, MP-2 medium and liquid culture system were able to maintain proliferation of embryogenic callus up to 83.1% with 3.23 multiplication rate. The best embryo germination up to 86.9% with 18.2 germinated embryos per clump within 21.3 days was determined on MPP-1 medium. While the best plantlet performances with 5 cm height of plantlets, 4.9 number of leaves, 2.8 number of roots, 2.6 cm root length, and 0.27 g plantlet fresh weight was obtained MPP-3 medium. With this propagation protocol, 3,000 - 4,000 plantlets/explant/year can be produced. Results of the study have high potential to be applied for in vitro propagation of Dendrobiums.

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Factors Affecting Plant Regeneration from Leaf Tissues of Buddleia Species

HortScience ◽

10.21273/hortsci.42.7.1670 ◽

2007 ◽

Vol 42 (7) ◽

pp. 1670-1673 ◽

Cited By ~ 9

Author(s):

Wenhao Dai ◽

Cielo Castillo

Keyword(s):

Shoot Regeneration ◽

Basal Medium ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Dark Treatment ◽

Regeneration Rate ◽

Factors Affecting ◽

In Vitro Shoots ◽

Leaf Tissues

The effects of genotype, basal medium, plant growth regulator (PGR), dark treatment, and antibiotics on shoot regeneration of two Buddleia cultivars, B. davidii ‘Potters Purple’ and Buddleia ‘Lochinch’, were investigated. In vitro shoots were regenerated from leaf tissues in either Murashige and Skoog (MS) or woody plant medium (WPM) media supplemented with benzyladenine (BA). In general, more shoots were regenerated in WPM medium than in MS medium. Dark treatment for 3 to 5 weeks dramatically increased shoot regeneration. Addition of indole-3-butyric acid (IBA) or naphthalene acetic acid (NAA) significantly enhanced the regeneration rate and shoots of each explant. The maximum regeneration rate (100%) of B. davidii ‘Potters Purple’ was achieved when cultured in WPM containing 5 μm BA plus 5 μm IBA. The maximum regeneration rate (98.4%) of Buddleia ‘Lochinch’ was found in WPM supplemented with 20 μm BA plus 4 μm IBA. Carbenicillin at 250 to 500 mg·L−1 and cefotaxime at 125 to 250 mg·L−1, individually or combined, promoted shoot regeneration. Interactions between genotype and medium or PGRs were found. In vitro shoots were easily rooted in half-strength MS medium with or without NAA. Rooted plants were transferred to potting mix and grown in the greenhouse. This research will facilitate genetic improvement and fast propagation of Buddleia species using biotechnology.

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In vitro propagation of five Alocasia species

Horticultura Brasileira ◽

10.1590/s0102-05362013000200006 ◽

2013 ◽

Vol 31 (2) ◽

pp. 210-215 ◽

Cited By ~ 2

Author(s):

Arvind Bhatt ◽

Christine Stanly ◽

Chan Lai Keng

Keyword(s):

Liquid Medium ◽

In Vitro Propagation ◽

Shake Flask ◽

Shoot Proliferation ◽

Ms Medium ◽

Shoot Height ◽

Shoot Number ◽

Significant Difference ◽

Shoot Tip Explants

The influence of cytokinin, N6-benzyladenine, on shoot proliferation of five Alocasia species (A. amazonica, A. cuprea, A. robusta, A. longiloba and A. chaii) was investigated. In vitro propagation of these species was established using shoot tip explants. Murashige & Skoog (MS) medium supplemented with different concentrations of BA (N6-benzyladenine) ranging from 0, 2, 5, 10 mg/L was then used to establish the optimum medium for shoot proliferation for all the species. MS medium supplemented with 2.0 mg/L BA was optimum for the shoot proliferation. All the tested species showed varying results for shoot number and shoot height. A comparison between agar-gelled medium and shake flask system using liquid medium was carried out to evaluate the shoot growth and proliferation for all the tested species. For A. amazonica, A. cuprea, A. robusta and A. longiloba, shake flask system using liquid medium of the same constituents stimulated more shoot proliferation as compared to agar-gelled medium. However, for A. chaii there was no significant difference. All the in vitro plantlets with well developed roots and leaves were successfully acclimatized with more than 90% survival rate.

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In Vitro Propagation, Huperzine A Content and Antioxidant Activity of Three Genotypic Huperzia serrata

Plants ◽

10.3390/plants10061112 ◽

2021 ◽

Vol 10 (6) ◽

pp. 1112

Author(s):

Yan Yang ◽

Liangfang Dai ◽

Decai Wu ◽

Limin Dong ◽

Yisheng Tu ◽

...

Keyword(s):

Antioxidant Activity ◽

High Performance ◽

In Vitro Propagation ◽

Huperzine A ◽

Induction Medium ◽

Naphthalene Acetic Acid ◽

Regeneration Rate ◽

Huperzia Serrata ◽

Chromatography Analysis

Huperzia serrata is a traditional herb and endangered Chinese medicinal material, which has attracted much attention due to its production of Huperzine A (HupA). In vitro propagation of H. serrata is considered a new way to relieve the resource pressure of H. serrata. In this study, three different genotypic wild H. serrata were used for in vitro propagation. Then, the antioxidant activity and the content of HupA in the regenerated H. serrata were investigated. The results showed the survival rate of the explant was increased to 25.37% when using multiple sterilization processes. The best induction medium for H. serrata was the Schenk and Hildebrandt (SH) medium supplemented with 0.5 mg·L−1 Naphthalene acetic acid (NAA) and 0.1 mg·L−1 2,4-Dichlorophenoxyacetic acid (2,4-D), where the regeneration rate of the explant was to 57.04%. The best proliferation medium was the SH medium with NAA (1.0 mg·L−1), as the biomass of in vitro tissue increased 164.17 ± 0.41 times. High-performance liquid chromatography analysis showed that the in vitro culture of three genotypes could produce HupA and the content of HupA was 53.90–87.17 µg·g−1. The antioxidant experiment showed that the methanol extract of in vitro H. serrata had higher antioxidant activity than that of wild H. serrata. This study provides a reliable in vitro H. serrata culture protocol and laid an important foundation for the antioxidant capacity of the thallus and the content of HupA.

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Influence of growth regulators and explants on shoot regeneration in carnation

Horticultural Science ◽

10.17221/1/2009-hortsci ◽

2009 ◽

Vol 36 (No. 4) ◽

pp. 140-146 ◽

Cited By ~ 4

Author(s):

J.K. Kanwar ◽

S. Kumar

Keyword(s):

Acetic Acid ◽

Shoot Regeneration ◽

Growth Regulators ◽

Dianthus Caryophyllus ◽

Naphthalene Acetic Acid ◽

Ms Medium ◽

Shoot Bud ◽

Dichlorophenoxy Acetic Acid ◽

Number Of Shoots

The influence of growth regulators, explants and their interactions on in vitro shoot bud formation from callus was studied in Dianthus caryophyllus L. The leaf and internode explants were cultured on Murashige and Skoog (MS) medium containing different concentrations of growth regulators. The highest callus induction was observed with 2 mg/l 2,4-dichlorophenoxy acetic acid (2,4-D) and 1 mg/l benzyl adenine (BA). Out of twenty seven shoot regeneration media tested, only 2 mg/l thidiazuron (TDZ) and zeatin alone or in combination with naphthalene acetic acid (NAA) and/or indole acetic acid (IAA) could differentiate calli. The highest average number of shoots was observed with 2 mg/l TDZ and 1 mg/l IAA. Significant differences were observed in calli producing shoots and number of shoots per callus in the explants of leaf and internode. The shoots were elongated and multiplied on MS medium supplemented with 1 mg/l BA and solidified with 1% agar. The shoots were rooted and hardened with 76% survival success in pots after six weeks of transfer to the pots.

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