Regeneration of Hemidesmus indicus (L.) R. Br. using in vitro nodes: an alternative method for efficient multiplication of shoots

In vivo nodes of Hemidesmus indicus (L.) R. Br. induced healthy multiple shoots with branching in our earlier studies and thus in the present study, potency of in vitro nodes to regenerate shoots was evaluated. In vitro nodes were excised from eight-week-old shoots and placed in Murashige and Skoog’s (MS) medium fortified with sucrose (3%) and different concentrations of 6-benzyladenine (BA) and kinetin (Kn). After eight weeks, optimum of 5.42 ± 0.36 shoots with 100% response were regenerated in medium supplemented with BA (10 µM) and Kn (5 µM). These healthy shoots were placed in full, half and quarter strengths of liquid MS medium fortified with sucrose (1%) and α-naphthaleneacetic acid (NAA, 1-25 µM) for rooting. Among all the strengths of MS medium, full strength MS medium having 8 µM NAA formed maximum of 3.42 ± 0.55 roots (91.67% response) within four weeks. The protocol is in continuation with earlier study and it was confirmed that a single in vivo nodal explant can regenerate around 385 healthy elongated shoots within 4 months, which will help in mass-propagation of the species.

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Direct Organogenesis from Rhizome Explants in Marsilea quadrifolia L.: A Threatened Fern Species

Advances in Biology ◽

10.1155/2015/639678 ◽

2015 ◽

Vol 2015 ◽

pp. 1-6 ◽

Cited By ~ 2

Author(s):

Mahipal S. Shekhawat ◽

M. Manokari

Keyword(s):

Multiple Shoots ◽

Direct Organogenesis ◽

Root Induction ◽

Ms Medium ◽

Full Strength ◽

Marsilea Quadrifolia ◽

Fern Species ◽

Half Strength

An efficient micropropagation protocol has been developed for Marsilea quadrifolia L. through direct organogenesis. The mature rhizomes were used as explants and successfully sterilized using 0.1% HgCl2 for the establishment of cultures. The multiple shoots were differentiated from the explants on Murashige and Skoog (MS) medium augmented with 6-benzylaminopurin (BAP). Full strength MS medium was reported to be effective for the induction of sporophytes from the rhizomes after four weeks of inoculation. Maximum response (96%) with average of 6.2 shoots (2.72 cm length) was achieved on full strength of MS medium augmented with 0.5 mg/L BAP in culture initiation experiments. The cultures were further proliferated in clusters (79.0±0.37 shoots per explant) with stunted growth on half strength MS medium supplemented with 0.25 mg/L BAP after four weeks. These stunted shoots were elongated (5.30 cm long) on half MS medium devoid of growth hormones. Root induction and proliferation (3.0–4.0 cm long) were observed after 4th subculture of sporophytes on hormone-free half strength MS medium. The rooted plantlets were hardened in the fern house for 4-5 weeks and transferred to the field with 92% survival rate. There were no observable differences in between in vivo grown and in vitro propagated plantlets in the field.

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Micropropagation and Comparative Study of Chemical Components of Essential Oils of in-vitro and in-vivo Grown Mentha spicata L.

Nepal Journal of Science and Technology ◽

10.3126/njst.v7i0.575 ◽

2007 ◽

Vol 7 ◽

pp. 71

Author(s):

B. R. Paudel ◽

B. Pant

Keyword(s):

Essential Oils ◽

Root Formation ◽

Shoot Culture ◽

Multiple Shoots ◽

Major Constituent ◽

Chemical Components ◽

Ms Medium ◽

Mentha Spicata

Axenic shoot culture of Mentha spicata L. was established from young shoots of the plant naturally growing in the field. Large number of multiple shoots were observed on MS media supplemented with NAA (0.5ppm) and BAP (1ppm) within eight weeks of culture. Extensive root formation was observed on MS medium supplemented with 1 ppm NAA. Essential oils from both in vitro and in vivo samples showed l- carvone as a major constituent, however, the in vitro samples showed higher concentration of menthol than in vivo samples. <i>Nepal Journal of Science and Technology</i> Vol. 7, 2006

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Shoot Proliferation, Callus Induction And Plant Regeneration In Tripsacum Laxum Nash

10.21203/rs.3.rs-880060/v1 ◽

2021 ◽

Author(s):

Yuping Xiong ◽

Jinhui Pang ◽

Kunlin Wu ◽

Jaime A. Teixeira Silva ◽

Xinhua Zhang ◽

...

Keyword(s):

Peat Soil ◽

Shoot Proliferation ◽

Multiple Shoots ◽

Dichlorophenoxyacetic Acid ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Axillary Shoots ◽

Rooting Medium ◽

2,4 Dichlorophenoxyacetic Acid

Abstract The peduncles of Tripsacum laxum Nash were used as explants to induce axillary shoots. Multiple shoots were proliferated on Murashige and Skoog (MS) medium to establish, for the first time, efficient shoot proliferation and plant in vitro regeneration systems. Optimal shoot proliferation medium was MS with 3.0 mg/L 6-benzyladenine (BA) and 0.2 mg/L α-naphthaleneacetic acid (NAA), resulting in a shoot proliferation coefficient of 11.0 within 45 d. Optimal rooting medium was MS with 0.1 mg/L NAA and/or 0.1 mg/L indole-3-butyric acid (IBA), inducing 100% root formation from shoots within 30 d. When young roots, leaf sheaths and shoot bases were used as explants, MS medium with 1.0 mg/L thidiazuron (TDZ) and 0.2 mg/L BA induced most shoots, with the least callus. Shoot bases induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L 2,4-dichlorophenoxyacetic acid (2,4-D), while leaf sheaths induced beige-white callus and shoots directly on MS medium with 1.0 mg/L TDZ and 0.2 mg/L BA. Rooted plantlets showed 99.3% survival when transplanted into a substrate of vermiculite: peat soil (1:3, v/v).

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IN VITRO PROPAGATION OF GUAVA (PSIDIUM GUAJAVA L.) FROM SEEDLING EXPLANTS

HortScience ◽

10.21273/hortsci.27.6.696e ◽

1992 ◽

Vol 27 (6) ◽

pp. 696e-696

Author(s):

Yaseen Mohamed-Yaseen ◽

Raymond J. Schnell ◽

Robert J. Knight ◽

T. L. Davenport

Keyword(s):

Shoot Growth ◽

Multiple Shoots ◽

Psidium Guajava ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Indolebutyric Acid ◽

Seedling Explants ◽

Cultivated Species ◽

Regenerated Plantlets

Guava (Psidium guajava L.) is an exceptional source of vitamin. C. It is also considered to be the most important cultivated species of the Myrtel family. Shoot tip and stem node were taken from seedling germinated in Murashige and Skoog medium (MS) and cultured in the same medium supplemented with 1-3mg/l benzylaminopurine (BA) and 0.1mg/l naphthaleneacetic acid (NAA) or 0.2-2mg/l thidiazuron (TDZ) and 0.1mg/l NAA. Multiple shoots (4-6) were obtained in 4-5 weeks from culture in 1-2mg/l BA and 0.1mg/l NAA, while TDZ caused abnormal shoot growth. Shoots were rooted successfully with 100% frequency in MS medium containing 2mg/l indolebutyric acid and further elongation of shoots was achieved in MS medium, supplemented with lg/l activated charcoal. Regenerated plantlets were successfully established in soil.

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Plant Production and Leaf Anatomy of Mertensia maritima (L.) Gray: Comparison of In Vitro Culture Methods to Improve Acclimatization

Horticulturae ◽

10.3390/horticulturae7050111 ◽

2021 ◽

Vol 7 (5) ◽

pp. 111

Author(s):

Andrea Copetta ◽

Miriam Bazzicalupo ◽

Arianna Cassetti ◽

Ilaria Marchioni ◽

Carlo Mascarello ◽

...

Keyword(s):

Solid Substrate ◽

Plant Production ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Temporary Immersion System ◽

Culture Methods ◽

Temporary Immersion ◽

Half Strength

Mertensia maritima is a commercially interesting herb with edible leaves and flowers, characterized by oyster flavor and taste. Plant propagation and traditional cultivation are challenging for this species. Therefore, the main purpose of the present study was to establish successful protocols aimed at ensuring oyster plant shoot propagation, rooting and in vivo acclimatization. Both micropropagation and rooting were tested, comparing the traditional in vitro solid substrate in jar vs. the liquid culture in a temporary immersion system (TIS) bioreactor (Plantform™). A Murashige and Skoog (MS) medium added with 4-µM thidiazuron (TDZ) and 1-µM α-naphthaleneacetic acid (NAA) was employed for micropropagation, while a half-strength MS medium supplemented with 4-µM indole−3-butyric acid (IBA) was used for rooting. Different acclimatization conditions in the greenhouse or in growth chamber were tested. Morphometric and microscopical analyses were performed on the oyster plant leaves at the propagation, rooting and acclimatization stages both in a jar and in a TIS. Micropropagation in a TIS allowed to obtain large shoots, while a great number of shoots was observed in the jar. M. maritima shoots rooted in TIS produced more developed roots, leaves with more developed waxy glands and well-formed stomata; moreover, the plants coming from the TIS showed the best acclimatization performances.

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Organogenesis and Ultrastructural Features ofIn VitroGrownCanna indicaL.

BioMed Research International ◽

10.1155/2016/2820454 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 1

Author(s):

Sharifah Nurashikin Wafa ◽

Rosna Mat Taha ◽

Sadegh Mohajer ◽

Noraini Mahmad ◽

Bakrudeen Ali Ahmed Abdul

Keyword(s):

Plant Regeneration ◽

Multiple Shoots ◽

Ms Medium ◽

Canna Indica ◽

Ultrastructural Studies ◽

Complete Plant ◽

Mother Plants ◽

Rhizome Explants

An efficient protocol for micropropagation ofCanna indicaL., an economically and pharmaceutically important plant, was standardized using rhizome explants, excised from two-month-old aseptic seedlings. Complete plant regeneration was induced on MS medium supplemented with 3.0 mg/L BAP plus 1.5 mg/L NAA, which produced the highest number of shoots (73.3 ± 0.5%) and roots (86.7 ± 0.4%) after 2 weeks. Furthermore, the optimum media for multiple shoots regeneration were recorded on MS enriched with 7.0 mg/L BAP (33.0 ± 0.5%). Plantlets obtained were transplanted to pots after two months and acclimatized in the greenhouse, with 75% survival. In addition, ultrastructural studies showed that rhizomes ofin vitrogrown specimens were underdeveloped compared to thein vivospecimens, possibly due to the presence of wide spaces. Meanwhile, the leaves ofin vivospecimens had more open stomata compared toin vitrospecimens, yet their paracytic stomata structures were similar. Hence, there were no abnormalities or major differences betweenin vitroregenerants and mother plants.

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Effects of Phytohormone and Regulators on Shoot Tip and Nodal Explants for In Vitro Shoot and Root Clonal Propagation of Vitex negundo L.

American Journal of Pure and Applied Biosciences ◽

10.34104/ajpab.021.065078 ◽

2021 ◽

pp. 65-78

Keyword(s):

Germplasm Conservation ◽

Multiple Shoots ◽

Shoot Tips ◽

Nodal Explants ◽

Nodal Segments ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Vitex Negundo ◽

Surface Sterilization

Medicinal plants are one of the most vital natural resources, but many of them are currently endangered due to habitat loss. Consequently, it is critical to emphasize the importance of using micropropagation techniques for mass propagation of plantlets on a commercial scale, in addition to germplasm conservation and distribution. Nodal explants and shoot tips were expunged from 15 days of the explant by aseptic seedlings, an effective, quick, and better in vitro plant regeneration procedure for Vitex negundo L. has been developed. The recent study was considered to develop an in vitro procedure for the regeneration of V. negundo L., a traditional medicinal plant. Nodal segments and shoot tips were cultivated on MS medium enhanced with numerous plant growth regulators. For multiple shoots and root regeneration, various cytokinins were examined. 6-benzyl-aminopurin (BAP), kinetin (Kin), and 1H-indole-3-butanoic acid (IBA) were all tested as a supplement to Murashige and Skoog (MS) medium including auxin phytohormone, such as Indole acetic acid (IAA) and 1-naphthaleneacetic acid (NAA). The furthermost effective surface sterilization treatment for explants of V. negundo has been found 0.1% HgCl2 for 8 minutes. In all treatments, multiple shoots were collected from shoot tips and nodal segments. In MS media added with 2.0mg/l BAP, the most shoots were seen in V. negundo. Furthermore, V. negundo regeneration shoots rooted effectively in half MS containing 1.0 mg/l IBA. Finally, proliferated plantlets were effectively adapted in soil, where they grew normally without morphological anomalies and had a survival rate of 92 percent.

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In Vitro Flowering and Fruiting of Capsicum fruitescens L.

HortScience ◽

10.21273/hortsci.30.1.130 ◽

1995 ◽

Vol 30 (1) ◽

pp. 130-132 ◽

Cited By ~ 17

Author(s):

Brent Tisserat ◽

Paul D. Galletta

Keyword(s):

Growth Regulators ◽

Agar Medium ◽

Fruit Size ◽

Fruit Production ◽

Shoot Tips ◽

Naphthaleneacetic Acid ◽

Ms Medium ◽

Immature Fruit

Flowering and fruit production were obtained from cultured shoot tips of `California Wonder', `Super Cayenne', and `Zippy' peppers grown in a liquid Murashige and Skoog (MS) medium without growth regulators. Thirty-day-old pepper shoot tips, derived from seedlings germinated sterilely, were cultured in a 6-liter polycarbonate container coupled to an automated plant culture system (APCS). Plants were given ten daily, 30-minute, compressed-air applications at 300 ml/minute. These shoots began flowering after an additional 60 to 90 days in culture and flowered continuously thereafter. About 5% to 10% of the flowers set fruit. Maximum fruit size obtained was ≈25% to 75% of the size of fruit produced on plants grown in vivo. In contrast, no flowering occurred from shoot tips grown in 25 × 150-mm culture tubes containing agar medium and subcultured every 8 weeks to fresh medium. Immature fruit excised from plantlets grown in the APCS were cultured separately on agar medium containing 0.0, 0.1, 0.3, or 1.0 mg BA/liter with and without 0.1 mg NAA/liter. Isolated fruit grew best on MS medium with BA only and poorest on medium without growth regulators. Chemical names used: N-(phenylmethyl)-1H-purine-6-amine (BA); 1-naphthaleneacetic acid (NAA).

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In vitro Embryogenesis Derived from Shoot Tips in Mass Propagation of Two Selected-Clones of Phalaenopsis

Notulae Scientia Biologicae ◽

10.15835/nsb839830 ◽

2016 ◽

Vol 8 (3) ◽

pp. 317-325 ◽

Cited By ~ 1

Author(s):

Budi WINARTO ◽

Kristina Dwi ATMINI ◽

Dedeh Siti BADRIAH ◽

Mega WEGADARA

Keyword(s):

Economic Value ◽

Shoot Tips ◽

Ms Medium ◽

Market Demand ◽

Mass Propagation ◽

Full Strength ◽

Half Strength ◽

Alternative Choice ◽

In Vitro Embryogenesis

Phalaenopsis is of high economic value and market demand in Indonesia; however, orchid products are mostly imported from other countries. ‘Kristina Dwi’ (KD) 69.274 and ‘Dedeh’ (D) 802.28 are two selected clones with high potential utilized and developed commercially. To support their commercialization, a reliable in vitro propagation protocol is essential. In the current study, an in vitro mass propagation protocol for KD 69.274 and D 802.28 clones was successfully established using shoot tips as explant sources. A high number of embryos, up to 8.2 embryos per explant, with 58.5% explant regeneration, and 3.5 regenerated-explants in average were regenerated from shoot tips of KD 69.274 clone cultured on half-strength Murashige and Skoog (MS) medium, with full strength micro, Fe-chellate and vitamin containing 0.5 mg/L thidiazuruon (TDZ) and 0.25 mg/L N6-benzyladenine (BA). The initial embryos were proliferated by culturing embryos individually on half-strength MS medium with 0.13 mg/L TDZ and 0.25 mg/L BA and resulted in high embryo regeneration up to 91.4%, with 10.2 embryos per explant and no embryo browning. The embryos were multiplied under periodical subcultures of 3 months each, resulting in gradual increasing number of embryos from the first subculture till the fifth subculture, with 23.6 embryos produced, then declined afterward. The embryos were easily germinated on half-strength MS medium with full strength of vitamin and hormone free, with 73.9% embryo germination and 14.9 germinated embryos. Healthy plantlets were stimulated on the same medium with 2 g/L activated charcoal (AC) and successfully acclimatized on Cycas rumphii bulk, with 88.3% survival plantlets. Finally, it can be summarized that a new in vitro mass propagation protocol, as new alternative choice for Phalaenopsis propagation, was successfully established.

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Plant Regeneration of Allium victorialis var. platyphyllum Makino via Organogenesis and Somatic Embryogenesis

HortScience ◽

10.21273/hortsci.31.4.628a ◽

1996 ◽

Vol 31 (4) ◽

pp. 628a-628

Author(s):

Hak-Tae Lim ◽

Eun-Ae Lee ◽

Won-Bae Kim

Keyword(s):

Somatic Embryogenesis ◽

Somatic Embryos ◽

Multiple Shoots ◽

Root Induction ◽

Ms Medium ◽

Mass Propagation ◽

Embryogenic Calli ◽

Wild Vegetable ◽

Shoot Tip Explants

This study was conducted to investigate the possibility of obtaining plantlets via somatic embryogenesis and organogenesis as means of in vitro mass propagation in Allium victorialis var. platyphyllum Makino, one of the most popular wild vegetable plants in Korea. Shoots formed directly when bulb explants of A. victorialis were cultured on MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin under 16 hours (light)/8 hours (dark) illumination. The use of leaf and shoot tip explants was not successful, largely due to explant senescence in the present of plant growth regulators. Embryogenic calli were obtained from the bulb explants of A. victorialis on MS medium supplemented with 0.2 mg·L–1 NAA, 0.2 mg·L–1 BAP, and 1.0 mg·L–1 picloram after 4–5 weeks of culture in the dark at 27°C. Upon transfer to shoot-induced MS medium containing 0.2 mg·L–1 NAA and 2.0 mg·L–1 zeatin, embryogenic calli gave rise to numerous somatic embryos, which subsequently developed into multiple shoots after 3 months of culture under 16 hours(light)/8 hours (dark) illumination. For root induction, regenerated shoots were transferred to MS medium added with 1.0 mg·L–1 NAA. Regenerants with well-developed roots were potted in an artificial soil mixture of vermiculite (1) and perlite (1).

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