High frequency plant regeneration from leaf culture of &lt;i&gt;Neolamarckia cadamba&lt;/i&gt;

Li, Jingjian; Deng, Zhili; Ouyang, Kang; Chen, Xiaoyang

doi:10.5511/plantbiotechnology.18.1119a

Cited by 14 publications

(15 citation statements)

References 26 publications

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“…The study demonstrates, for the first time, an improved, reproducible and highly efficient buds/shoots regeneration protocol of N. cadamba was developed in both cotyledonary petiole and hypocotyl explants, used the cytokinin TDZ via an intermediate callus formation. Obviously, in terms of proliferation multiple and potential genetic transformation ability, this in vitro regeneration strategy is superior to our previous research 20 . In addition, this is the first report of genetically sustainable proliferation via cotyledonary petiole and hypocotyl explants in N. cadamba.…”

Section: Resultsmentioning

confidence: 74%

High frequency regeneration of plants via callus-mediated organogenesis from cotyledon and hypocotyl cultures in a multipurpose tropical tree (Neolamarkia Cadamba)

Huang

Wei

Zhai

et al. 2020

Sci Rep

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In this works, a simple, efficient and repeatable protocol was developed for in vitro regeneration via callus-mediated organogenesis of Neolamarkia Cadamba using cotyledonary petioles and hypocotyls. Effects of basal medium, plant growth regulators, the types and age of explant on the formation of adventitious buds/shoots were studied. Meanwhile, histological analysis for early ontogenic stages and genetic stability assessment by flow cytometry were investigated. Our investigation demonstrated that, compared with 6-benzyladenine (BA), N 6-(2-isopentenyl) adenine (2-ip), Thidiazuron (TDZ) was the optimal cytokinin for buds/shoots induction on cotyledon and hypocotyl explants. Douglas-fir and sugar pine medium (DCR) supplemented with 22.7 μM TDZ and 0.27 μM α-naphthalene acetic acid (NAA) was most effective on bud induction, with the highest bud-induction rate and numbers of buds on cotyledon and hypocotyl explants. The available shoot per explant hit 35.2 when the induced callus sub-cultured to a medium without TDZ. It was found that TDZ could promote induction of the callus and the buds, however, continuous exposure beyond 4 weeks of supplemented high concentration (exceed 11.35 μM), TDZ was harmful to the proliferation and growth of buds/shoots. DCR appeared more efficiency than Murashige and Skoog medium (MS), Woody Plant medium (WPM), anther culture of cereal crops medium (n 6) on bud induction. Age of cotyledon and hypocotyl explants in 20-day to 25day was most beneficial to adventitious buds/shoots formation. Histological investigation confirmed that the buds originated from the wounded incisions of cotyledonary petiole and hypocotyl fragments, with callus formation. The regeneration plantlets were successfully acclimatized in greenhouse, yielded above 95% survival rate in field, exhibited normal morphology and growth characteristics. The analysis of flow cytometry on N. cadamba indicated no variation in the ploidy levels between the regenerated plantlets and the donor trees. The developed procedure can be used for mass production, germplasm exchange and transgenic studies to improve the resistance of the species via Agrobacterium-mediated. Neolamarckia cadamba (commonly known as burflower tree, kadamba tree or kadamb) is a tropical tree of great economic importance. First, N. cadamba trees grow fast, and their wood is good for pulp production, furniture, and other construction purposes 1. Second, N. cadamba has been traditionally used as medicine to treat diseases such as dysentery, fever and snake bites 2,3. Now, from its bark, leaves, and flowers people extract phytochemical compounds, including monoterpenoid, triterpenoid saponin, and ethylene glycol 4-9 , alkaloid 10 , cadambine 7,11 , and use them to treat hyperglycemia 12 , hyperlipidemia 13,14 , hypertension 15 , and wound and skin diseases 16. Third,

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Section: Resultsmentioning

confidence: 74%

High frequency regeneration of plants via callus-mediated organogenesis from cotyledon and hypocotyl cultures in a multipurpose tropical tree (Neolamarkia Cadamba)

Huang

Wei

Zhai

et al. 2020

Sci Rep

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“…Morphological examination revealed that the application of NAA produced EC specifically, which was characterized by a compact shape, pale green color, and the presence of small, granular structures. In general, EC may proceed down several morphogenic processes: somatic embryogenesis, nodular embryogenic structures (NESs), or shoot organogenesis [ 34 ]. The regeneration of shoots from leaf explants generally occurs through the induction of EC, and often requires the presence of both auxins and cytokinins [ 35 ].…”

Section: Discussionmentioning

confidence: 99%

Development of a Highly Efficient Shoot Organogenesis System for an Ornamental Aeschynanthus pulcher (Blume) G. Don Using Leaves as Explants

Yang

Yuan

et al. 2022

Plants

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Aeschynanthus pulcher (Blume) G. Don, the “lipstick plant” is a prized ornamental plant with distinctive flowers. Here, we introduce a novel in vitro regeneration method for A. pulcher using leaf explants and an optimized combination of phytohormone plant growth regulators (PGRs). The optimal conditions for shoot regeneration included 1 mg L−1 polyvinyl pyrrolidone (PVP) plus 3 mg L−1 thidiazuron (TDZ), inducing a response rate of 82.4% and a shoot/explant ratio of 38.6. When the Murashige and Skoog (MS) medium contained indole-3-butyric acid (IBA) alone, leaves first differentiated into adventitious roots and then adventitious shoots. Leaves cultured on MS medium containing 1 g L−1 PVP, 3 mg L−1 TDZ, 5 mg L−1 casein, and 0.1 mg L−1 α-naphthaleneacetic acid (NAA) for 30 d exhibited the highest embryogenic callus (EC) induction rate (95.6%). The optimal shoot proliferation coefficient (21.5) was obtained when shoots derived from EC were cultured on the same medium as that used for EC induction for 5 weeks. The most effective medium for rooting of elongated shoots was MS medium containing 1 g L−1 PVP, 5 mg L−1 casein, 3 mg L−1 6-benzyladenine (BA), and 0.1 mg L−1 NAA, and the number of roots reached 18.8. The regenerated plants grown in a greenhouse had 100% survival following one week of hardening. Overall, our effective and efficient propagation method should result in shortened culture periods and reduced production costs, allowing for the future selective breeding and genetic improvement of A. pulcher.

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“…0.1%) of bioactive 3dihydrocadambine and cadambine in the bark and leaves. Given that we have established a highly efficient regeneration system of N. cadamba (Li et al, 2019), the genomic information presented here will greatly facilitate the elucidation of the cadambine synthetic pathway and the development of tools for enhancing bioactive productivity by metabolic engineering in microbes or by molecular breeding in plants.…”

Section: Discussionmentioning

confidence: 99%

Chromosome‐level assembly of the Neolamarckia cadamba genome provides insights into the evolution of cadambine biosynthesis

Zhao

Ouyang

et al. 2021

The Plant Journal

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Neolamarckia cadamba (Roxb.), a close relative of Coffea canephora and Ophiorrhiza pumila, is an important traditional medicine in Southeast Asia. Three major glycosidic monoterpenoid indole alkaloids (MIAs), cadambine and its derivatives 3b-isodihydrocadambine and 3b-dihydrocadambine, accumulate in the bark and leaves, and exhibit antimalarial, antiproliferative, antioxidant, anticancer and anti-inflammatory activities. Here, we report a chromosome-scale N. cadamba genome, with 744.5 Mb assembled into 22 pseudochromosomes with contig N50 and scaffold N50 of 824.14 Kb and 29.20 Mb, respectively. Comparative genomic analysis of N. cadamba with Co. canephora revealed that N. cadamba underwent a relatively recent whole-genome duplication (WGD) event after diverging from Co. canephora, which contributed to the evolution of the MIA biosynthetic pathway. We determined the key intermediates of the cadambine biosynthetic pathway and further showed that NcSTR1 catalyzed the synthesis of strictosidine in N. cadamba. A new component, epoxystrictosidine (C27H34N2O10, m/z 547.2285), was identified in the cadambine biosynthetic pathway. Combining genome-wide association study (GWAS), population analysis, multi-omics analysis and metabolic gene cluster prediction, this study will shed light on the evolution of MIA biosynthetic pathway genes. This N. cadamba reference sequence will accelerate the understanding of the evolutionary history of specific metabolic pathways and facilitate the development of tools for enhancing bioactive productivity by metabolic engineering in microbes or by molecular breeding in plants.

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High frequency plant regeneration from leaf culture of <i>Neolamarckia cadamba</i>

Cited by 14 publications

References 26 publications

High frequency regeneration of plants via callus-mediated organogenesis from cotyledon and hypocotyl cultures in a multipurpose tropical tree (Neolamarkia Cadamba)

High frequency regeneration of plants via callus-mediated organogenesis from cotyledon and hypocotyl cultures in a multipurpose tropical tree (Neolamarkia Cadamba)

Development of a Highly Efficient Shoot Organogenesis System for an Ornamental Aeschynanthus pulcher (Blume) G. Don Using Leaves as Explants

Chromosome‐level assembly of the Neolamarckia cadamba genome provides insights into the evolution of cadambine biosynthesis

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