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.)

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 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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In vitro Plant Regeneration of Four Local Varieties of Chickpea (Cicer arietinum L.) Grown in Bangladesh

Bangladesh Journal of Scientific and Industrial Research ◽

10.3329/bjsir.v46i3.9047 ◽

1970 ◽

Vol 46 (3) ◽

pp. 379-384 ◽

Cited By ~ 2

Author(s):

TA Banu ◽

RH Sarker ◽

MI Hoque

Keyword(s):

In Vitro Regeneration ◽

Multiple Shoots ◽

Mature Embryo ◽

Multiple Shoot ◽

Direct Organogenesis ◽

Ms Medium ◽

Regeneration System ◽

Best Response ◽

Multiple Shoot Regeneration

In vitro regeneration system was developed through direct organogenesis from decapitated mature embryo explants of locally grown four chickpea varieties, namely, Barichhola-4, Hyprochhola, Binachhola-3 and Binachhola-4. Best response towards multiple shoot regeneration was obtained on MS medium supplemented with 0.5 mg/l BAP, 0.5 mg/l Kn, 0.2 mg/l NAA along with double concentrations of CaCl2 and NH4NO3. However good shoot health and expanded leaf was found on MS medium containing 1.0 mg/l kn. Apart from this, few experiments were conducted with decapitated embryo attached cotyledon. Using this explants highest number of multiple shoots were obtained on MSB medium containing 4× micronutrients of MS medium with 3.0 mg/l BAP and 0.04 mg/l NAA in all four varieties. Shoots regenerated on 1.0 mg/l kn supplemented medium showed good response towards rooting on MS medium supplemented with 0.2 mg/l IBA in all four varieties. It was observed that micrografting is an alternative technique to in vitro rooting in chickpea. Key words: In vitro regeneration; Decapitated embryo; Chickpea. DOI: http://dx.doi.org/10.3329/bjsir.v46i3.9047 BJSIR 2011; 46(3): 379-384

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In vitro Regeneration System of Couroupita guianensis Using Cotyledonary Nodes

Asian Journal of Biological Sciences ◽

10.3923/ajbs.2019.412.422 ◽

2019 ◽

Vol 12 (3) ◽

pp. 412-422 ◽

Cited By ~ 1

Author(s):

Shiny Arokiamary Pushparaj ◽

Vinoth Alphonse ◽

Ravindhran Ramalingam

Keyword(s):

In Vitro Regeneration ◽

Regeneration System ◽

Cotyledonary Nodes ◽

Couroupita Guianensis

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In vitro Plant Regeneration from Seedling-derived Explants of two Cultivars of White Jute (Corchorus capsularies L.)

Plant Tissue Culture and Biotechnology ◽

10.3329/ptcb.v19i2.5437 ◽

2010 ◽

Vol 19 (2) ◽

pp. 199-206

Author(s):

Rakha Hari Sarker ◽

Bivas Kumar Sarkar ◽

M. I. Hoque ◽

M. I. Hoque

Keyword(s):

Plant Regeneration ◽

In Vitro Regeneration ◽

Cotyledonary Node ◽

Direct Organogenesis ◽

In Vitro Plant Regeneration ◽

Regeneration System ◽

Shoot Initiation ◽

Corchorus Capsularis ◽

In Vitro Plant

An in vitro regeneration system for two varieties of white jute (Corchorus capsularis L.) namely, BJC-7370 and BJC-83 was developed. The regeneration protocol was based upon direct organogenesis from seedling-derived explants such as cotyledon with petioles, cotyledonary node. Shoot regeneration was achieved from cotyledon with petiole and cotyledonary node through use of MS supplemented with 0.5 mg/l BAP and 1.0 mg/l IAA for BJC-7370 while for BJC-83 shoot initiation was obtained on 1.25 mg/l BAP and 0.25 mg/l NAA using the same explants. Elongation of shoots was achieved on MS containing 0.2 mg/l BAP for the two said jute varieties. Regenerated excised shoots developed effective in vitro root system on half strength of MS supplemented with 0.3 mg/l IBA for both the varieties. The in vitro grown plantlets were transferred to soil for acclimation. These plants grew up to maturity, flowered and produced seeds identical to the control plants. Key words: Corchorus capsularies, In vitro plant regeneration, seedling explants D.O.I. 10.3329/ptcb.v19i2.5437 Plant Tissue Cult. & Biotech. 19(2): 199-206, 2009 (December)

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Callus induction and adventitious bud differentiation of Cyclocodon lancifolius (Roxb.) Kurz

Botanical Sciences ◽

10.17129/botsci.2609 ◽

2020 ◽

Vol 98 (4) ◽

pp. 534-544

Author(s):

Yin-Kai Xi ◽

Ye Wang ◽

Biao Zeng ◽

Heng-Yu Huang ◽

Wu-Dei Yang

Keyword(s):

Callus Induction ◽

In Vitro Regeneration ◽

Culture Method ◽

Adventitious Shoot ◽

Adventitious Bud ◽

Ms Medium ◽

Regeneration System ◽

Edible Plant ◽

Wild Resources

Background: Cyclocodon lancifolius (Roxb.) Kurz is a perennial medicinal and edible plant with a huge potential economic value. The wild resources of this plant are gradually scarce by the serious destruction of the habitat and the limitations of sexual reproduction. This is the first attempt to establish an in vitro reproductive system for the species. Hypothesis: The suitable plant regulator types and its mass concentration range, combined with the explants, can induce the development of plants at various stages. We expected to establish an in vitro regeneration system of C. lancifolius based on these factors. Species studied: Cyclocodon lancifolius Study site and years of study: Yunnan Breeding and Cultivation Research and Development Center of Endangered and Daodi Chinese Medicinal Materials, Yunnan University of Chinese Medicine, from 2017 to 2019. Methods: The plant regeneration of C. lancifolius was established by single factor, L9 (34) orthogonal and complete combination experiments. Results: Stem segments were the best explants for callus induction on MS medium containing 0.05 mg·L-1 KT, 0.5 mg·L-1 6-BA and 0.5 mg·L-1 NAA. MS medium with 0.2 mg·L-1 TDZ and MS medium were used alternately as the culture method to conduct differentiation and proliferation of adventitious shoot. The optimal protocol for the rooting was MS medium combined with 0.1 mg·L-1 6-BA and 1.0 mg·L-1 NAA. Conclusions: A rapid propagation system of C. lancifolius was established which provided a possible solution for the protection of wild resources and artificial cultivation.

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Direct in vitro Regeneration of Arachis Hypogaea L.(Jl-24) and Study of Salt Tolerance

Indian Journal Of Applied Research ◽

10.15373/2249555x/aug2013/17 ◽

2011 ◽

Vol 3 (8) ◽

pp. 52-54 ◽

Cited By ~ 1

Author(s):

Bhagya Lakshmi Jyothi Kusuma ◽

◽

Sharmila Begum S

Keyword(s):

Salt Tolerance ◽

Arachis Hypogaea ◽

In Vitro Regeneration ◽

Arachis Hypogaea L

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Intensification de la régénération du pois (Pisum sativum L.), par le thidiazuron, via la formation de structures caulinaires organogènes

Canadian Journal of Botany ◽

10.1139/b97-053 ◽

1997 ◽

Vol 75 (3) ◽

pp. 492-500 ◽

Cited By ~ 6

Author(s):

Delphine Popiers ◽

Frédéric Flandre ◽

Brigitte S. Sangwan-Norreel

Keyword(s):

Pisum Sativum ◽

In Vitro Regeneration ◽

Naphthaleneacetic Acid ◽

Cotyledonary Nodes ◽

Pisum Sativum L ◽

Embryo Axes ◽

Initial Medium ◽

Second Wave ◽

Mature Seeds

In vitro regeneration of pea (Pisum sativum L.), a regeneration recalcitrant legume, was optimised using thidiazuron. Buds were initiated from the meristems of the cotyledonary nodes of embryo axes, isolated from mature seeds, and subcultured on Murashige and Skoog medium supplemented with 13.3 μM 6-benzylaminopurine, 16.1 μM α-naphthaleneacetic acid, and 0.2 μM 2,3,5-triiodobenzoic acid. Proliferation of buds was preceded by the formation of white nodular-like protrusions. These structures were cut transversally in fine slices and subcultured on the same medium or in presence of thidiazuron that produces a second wave of secondary budding. The best results (90–110 buds per expiant) were obtained with 10 μM thidiazuron. The capacity of regeneration was genotype independent and reproducible. Buds elongated on the initial medium, then formed roots in presence of 5.37 μM α-naphthaleneacetic acid. and developed into viable plants. Key words: Pisum sativum L., regeneration, meristems, embryo axes, thidiazuron.

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