In vitro regeneration of mulberry plants from seedling explants of Morus indica cv. G4 through direct organogenesis

In this present work, an in-vitro regeneration protocol for Crescentia alata (C. alata) was developed using various explants on Murashige and Skoog (MS) medium augmented with different concentrations and combinations of plant growth regulators (PGRs) for direct and indirect regeneration. The direct organogenesis was established from nodes and internodes on MS medium supplemented with cytokinins and auxins. The indirect organogenesis via callus phase was obtained from leaf, nodes and internodes on MS medium supplemented with different concentrations of PGRs. The high frequency shoot organogenesis were achieved directly from nodal explants were cultured on MS medium supplemented with 3.0 mg/L BAP+0.5 mg/L KIN +1.0 mg/L NAA. Indirect organogenesis callogenic frequency was optimized at the concentration of MS medium containing 1.0 mg/L BAP + 5.0 mg/L IAA. The callus was obtained from all the explants were used, among these explants internodal explants gave best result on MS medium supplemented with different concentrations of cytokinins and auxins for indirect organogenesis experiment. Indirect organogenesis the highest number of shoot regeneration was obtained in MS Basal Medium with 4.0 mg/L BAP + 0.5 mg/L KIN + 2.0 mg/L NAA from internodal explants. For root formation the regenerative shoots which were sub cultured on MS medium containing different ratios of auxins. The rooted plantlets were transferred successfully to the pots containing sterilized soil and were successfully hardened at greenhouse condition for 20 days then exposed to the natural environment. This is the first successful micropropagation report of an efficient and rapid in-vitro clonal propagation protocol for C. alata by direct and indirect shoot organogenesis through various explants, which can be employed for conservation of this important medicinal tree species as well as the utilization of an biologically important active biomolecules. This protocol can be very useful to obtain plants from various explants, without the requirement of meristematic regions, enabling the obtainment of a higher number of plants in short period.

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Histological studies of in vitro regeneration induced in leaf explants of Nymphoides cristatum (Roxb) O.Kuntze – An aquatic medicinal plant

International Journal of Life Sciences ◽

10.3126/ijls.v3i0.2370 ◽

1970 ◽

Vol 3 ◽

Author(s):

Mallapa Hanumanthu Niranjan ◽

Mysore Shankar Sudarshana

Keyword(s):

Medicinal Plant ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Direct Organogenesis ◽

Ms Medium ◽

Free Medium ◽

Histological Studies ◽

Leaf Histology ◽

Benzyl Amino Purine

The objective of this work was to study the histological events related to the regeneration process of a medicinal plant, Nymphoides cristatum (Roxb). Leaf explants were cultured on MS medium supplemented with 0.5 mgl-1 of 6-benzyl amino purine (BAP). About 90% of explants gave rise to shoots after 15 days of incubation. The histological studies showed that the regeneration originated directly from parenchymatous cells and direct organogenesis after 20 days of culture could be observed. Buds and roots were found completely differentiated after 40 days of culture and number of shoots per explants was 30. Micorshoots were rooted in hormone - free medium and the plants obtained grew in artificial pond under green house conditions. Key words: Leaf, Histology, in vitro regeneration, Nymphoides cristatum. DOI: 10.3126/ijls.v3i0.2370

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In Vitro Regeneration of Valencia-type Peanut (Arachis hypogaea L.) from Cultured Petiolules, Epicotyl Sections and Other Seedling Explants1

Peanut Science ◽

10.3146/i0095-3679-19-2-5 ◽

1992 ◽

Vol 19 (2) ◽

pp. 82-87 ◽

Cited By ~ 25

Author(s):

Ming Cheng ◽

David C. H. Hsi ◽

Gregory C. Phillips

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Shoot Formation ◽

Direct Organogenesis ◽

Naphthaleneacetic Acid ◽

Regeneration System ◽

Arachis Hypogaea L ◽

Wide Range ◽

Cultivated Peanut

Abstract This study evaluated plant development via direct organogenesis from in vitro-cultured young seedling tissues of cultivated peanut, especially the valencia-type peanut. Complete plants were regenerated from in vitro-cultured petiolule-with-blade-attached explants, leaflet segments, and epicotyl andpetiole sections. Multiple shoots arose on Murashige and Skoog medium (MS) supplemented with 6-benzylaminopurine (BA) (5–25 mg/L) plus 1-naphthaleneacetic acid (NAA) (0.5–3 mg/L). After 30 d culture on 25 mg/L BA + 1 mg/L NAA, 1.6 buds or shoots/explant were regenerated from the petiolule-with-blade-attached explants. Comparable numbers of shoots were obtained from epicotyl sections of the first node region of the seedling after 60 d culture using 10 mg/L BA + 1 mg/L NAA. Leaflet segments and petiole sections were less responsive for shoot formation. Excised shoots developed roots in vitro upon transfer for 15 d to MS medium supplemented with NAA at 1 mg/L. Plantlets were transferred to soil and grown in a greenhouse to maturity. A wide range of cultivated peanut genotypes was evaluated for organogenic responsiveness, using the petiolule-with-blade-attached explant source. Only valencia-type cultivars, or a hybrid derivative with a Valencia background, were responsive with this regeneration system.

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Establishment of an efficient alfalfa regeneration system via organogenesis and the co-expression of Arabidopsis SOS genes improves salt tolerance in transgenic alfalfa (Medicago sativa L.)

Canadian Journal of Plant Science ◽

10.1139/cjps-2018-0105 ◽

2019 ◽

Vol 99 (3) ◽

pp. 348-363

Author(s):

Jing Wang ◽

Xing Xu ◽

Dongmei Ma

Keyword(s):

Salt Tolerance ◽

Transgenic Plants ◽

In Vitro Regeneration ◽

Plant Genome ◽

Direct Organogenesis ◽

Adventitious Bud ◽

Transcriptional Level ◽

Regeneration System ◽

Cotyledonary Nodes

The Salt Overly Sensitive (SOS) signal transduction pathway is pivotal in Na+ efflux and facilitates ion transport and homeostasis for improved salt tolerance in plants. Ten alfalfa varieties were used as experimental materials and two alfalfa regeneration systems were established and optimized. Cotyledons and hypocotyls were initially used as explants to induce embryogenic callus via the indirect production of somatic embryos to establish a callus acceptor system. Cotyledonary nodes were used as explants to induce adventitious bud formation via direct organogenesis, thereby establishing an in vitro regeneration system that could be used for the genetic transformation. Agrobacterium-mediated transformation of the cotyledonary nodes of the alfalfa ‘Golden Empress b’ was used to generate 25 independent sources of transformed plants exhibiting herbicide tolerance. Four of the positive transgenic plants were randomly selected for southern blot analysis, and three hybridization signals with one or two copies were detected. Reverse transcription polymerase chain reaction showed that the Bialaphos resistance (Bar) and SOS1 genes were expressed in transgenic plants and that multiple exogenous salt-tolerant genes were integrated into the transgenic plant genome and expressed at the transcriptional level. The overexpression of Arabidopsis SOS genes in alfalfa conferred a high degree of salinity tolerance, enhanced plant growth, lowered the accumulation of Na+, increased the accumulation of K+ in the leaves, and altered physiological and biochemical parameters in response to salt stress.

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In vitro regeneration of Carlina acaulis subsp. simplex from seedling explants

Acta Physiologiae Plantarum ◽

10.1007/s11738-008-0252-5 ◽

2009 ◽

Vol 31 (3) ◽

pp. 445-453 ◽

Cited By ~ 9

Author(s):

Alina Trejgell ◽

Grażyna Dąbrowska ◽

Andrzej Tretyn

Keyword(s):

In Vitro Regeneration ◽

Seedling Explants

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In vitro regeneration of Alstroemeria cv. ‘Yellow King’ by direct organogenesis

Plant Cell Tissue and Organ Culture (PCTOC) ◽

10.1007/s11240-005-9020-z ◽

2005 ◽

Vol 84 (2) ◽

pp. 189-198 ◽

Cited By ~ 5

Author(s):

Martha E. Pedraza-Santos ◽

Ma. Cristina López-Peralta ◽

Víctor A. González- Hernández ◽

E. Mark Engleman-Clark ◽

Prometeo Sánchez-García

Keyword(s):

In Vitro Regeneration ◽

Direct Organogenesis

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Development of In-Vitro Regeneration Protocol From Seedling Explants of Populus Gamblei Dode: A Difficult to Propagate Endemic Tree Species of Eastern Himalayas

10.21203/rs.3.rs-910342/v1 ◽

2021 ◽

Author(s):

Puran Chandra ◽

Ajay Thakur ◽

K P Mohapatra ◽

T S Mehra ◽

Arti Kunwar ◽

...

Keyword(s):

Tree Species ◽

In Vitro Regeneration ◽

Subtropical Forest ◽

Nodal Segment ◽

Eastern Himalayas ◽

First Report ◽

Percent Survival ◽

Regenerated Plants ◽

Seedling Explants

Abstract This is the first report of in-vitro regeneration of Populus gamblei, an endemic and rare tree of montane to subtropical forest of Eastern Himalayas. Nodal segment from coppice shoot of young headed back seedling resulted in 100 % sprouting in MS + 1.0 mg L-1 BAP. Each micro shoot proliferated and gave 7.06 micro shoots on MS + 0.50 mg L-1 BAP + 0.10 mg L-1 IBA. In vitro multiplied shoots produced 100 % rooting in ½ MS + 0.5 mg L-1 NAA. In-vitro regenerated plants recorded 80 percent survival during hardening.

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Cytokinin-Based Tissue Cultures for Stable Medicinal Plant Production: Regeneration and Phytochemical Profiling of Salvia bulleyana Shoots

Biomolecules ◽

10.3390/biom11101513 ◽

2021 ◽

Vol 11 (10) ◽

pp. 1513

Author(s):

Izabela Grzegorczyk-Karolak ◽

Katarzyna Hnatuszko-Konka ◽

Marta Krzemińska ◽

Monika A. Olszewska ◽

Aleksandra Owczarek

Keyword(s):

Medicinal Plant ◽

Growth Parameters ◽

In Vitro Regeneration ◽

Leaf Explants ◽

Plant Production ◽

Bioactive Metabolites ◽

Direct Organogenesis ◽

Genetic Profile ◽

Adventitious Shoot Formation

Salvia bulleyana is a rare Chinese medicinal plant that due to the presence of polyphenols lowers the risk of some chronic diseases especially those related to the cardiovascular system. The present study examines the organogenic competence of various combinations and concentrations of plant growth regulators to develop an efficient protocol for in vitro regeneration of S. bulleyana via leaf explants, maintaining the high production of active constituents. The purpose of the study was also to assess the possibilities of using a cytokinin-based regeneration to effectively produce therapeutic compounds. The adventitious shoot formation was observed through direct organogenesis on media with purine derivatives (meta-topolin, mT and benzylaminopurine, BAP), and through indirect organogenesis on media with urea derivatives (tidiazuron, TDZ and forchlorfenuron, CPPU). The highest regeneration frequency (95%) with 5.2 shoots per explant was obtained on leaves cultured on Murashige and Skoog (MS) medium containing 0.1 mg/L naphthalene-1-acetic acid (NAA) and 2 mg/L BAP. Following inter simple sequence repeat (ISSR) marker-based profiling, the obtained organogenic shoot lines revealed a similar banding pattern to the mother line, with total variability of 4.2–13.7%, indicating high level of genetic stability. The similar genetic profile of the studied lines translated into similar growth parameters. Moreover, HPLC analysis revealed no qualitative differences in the profile of bioactive metabolites; also, the total polyphenol content was similar for different lines, with the exception of the shoots obtained in the presence of CPPU that produced higher level of bioactive compounds. This is the first report of an effective and rapid in vitro organogenesis protocol for S. bulleyana, which can be efficiently employed for obtaining stable cultures rich in bioactive metabolites.

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