In vitro regeneration, Agrobacterium-mediated transformation, and genetic assay of chalcone synthase in the medicinal plant Echinacea pallida

Wang, Hsin-Mei; Jeng, Shih‐Tong; To, Kin‐Ying

doi:10.1007/s11240-017-1208-5

Cited by 7 publications

(8 citation statements)

References 61 publications

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“…In brief, incomplete integration of T-DNA into the plant chromosome during the transformation process was found in transgenic plants OD24 and FAA11. Previously, using the transformation vector pCHS, which carries the Petunia chalcone synthase (chs) and nptII genes, we also observed incomplete integration of T-DNA in several plant species, including B. pilosa [30], the floricultural plant Cleome spinosa [31], and the medicinal plant Echinacea pallida [32]. The loss of one of the two transgenes within the same T-DNA has been clearly demonstrated in transgenic wheat [33] and rice [34].…”

Section: Discussionmentioning

confidence: 98%

“…Oligonucleotide sequences of specific FAA primers (FAA-F, FAA-R, cFAA-f1, cFAA-f2, cFAA-r1, cFAA-r2) and specific OD primers (OD-F, OD-R, cOD-f1, cOD-f2, cOD-r1, cOD-r2) are listed in Supplementary Table S1. Protocols for gel purification, cloning into pGEM-T Easy Vector (Promega, Madison, WI, USA), and transformation into Escherichia coli JM109 competent cells were previously described [32]. Plasmid DNA was isolated and then sequenced.…”

Section: Cloning Of Full-length Cdnasmentioning

confidence: 99%

“…Total RNA was isolated from the green leaves of WT and transgenic plants by CTAB [47]. Quantitative real-time PCR (qRT-PCR) analysis was carried out as previously described [32]. Specific primer sets for the amplification of DNA fragments for FAA (187 bp), OD (234 bp), and ribosomal protein L2 (100 bp; internal control) from B. pilosa are listed in Supplementary Table S1.…”

Section: Quantitative Real-time Pcr Analysismentioning

confidence: 99%

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Transformation and Characterization of Δ12-Fatty Acid Acetylenase and Δ12-Oleate Desaturase Potentially Involved in the Polyacetylene Biosynthetic Pathway from Bidens pilosa

Chen

Hsieh

Tsai

et al. 2020

Plants

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Bidens pilosa is commonly used as an herbal tea component or traditional medicine for treating several diseases, including diabetes. Polyacetylenes have two or more carbon–carbon triple bonds or alkynyl functional groups and are mainly derived from fatty acid and polyketide precursors. Here, we report the cloning of full-length cDNAs that encode 12-fatty acid acetylenase (designated BPFAA) and 12-oleate desaturase (designated BPOD) from B. pilosa, which we predicted to play a role in the polyacetylene biosynthetic pathway. Subsequently, expression vectors carrying BPFAA or BPOD were constructed and transformed into B. pilosa via the Agrobacterium-mediated method. Genomic PCR analysis confirmed the presence of transgenes and selection marker genes in the obtained transgenic lines. The copy numbers of transgenes in transgenic lines were determined by Southern blot analysis. Furthermore, 4–5 FAA genes and 2–3 OD genes were detected in wild-type (WT) plants. Quantitative real time-PCR revealed that some transgenic lines had higher expression levels than WT. Western blot analysis revealed OD protein expression in the selected transformants. High-performance liquid chromatography profiling was used to analyze the seven index polyacetylenic compounds, and fluctuation patterns were found.

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Section: Discussionmentioning

confidence: 98%

Section: Cloning Of Full-length Cdnasmentioning

confidence: 99%

Section: Quantitative Real-time Pcr Analysismentioning

confidence: 99%

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Transformation and Characterization of Δ12-Fatty Acid Acetylenase and Δ12-Oleate Desaturase Potentially Involved in the Polyacetylene Biosynthetic Pathway from Bidens pilosa

Chen

Hsieh

Tsai

et al. 2020

Plants

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“…Still, uniform flowering, equal pick length, higher essential oil genotypes and additional yield using suitable agrotechnologies are some of the crucial areas that require attention in future German chamomile studies. Several researchers aim to genetic engineer the growth-process of the plant to improve secondary metabolites [ 107 , 108 ].…”

Section: Genetic Improvementmentioning

confidence: 99%

A Comprehensive Review on Biology, Genetic Improvement, Agro and Process Technology of German Chamomile (Matricaria chamomilla L.)

Chauhan

Singh

Kumar

et al. 2021

Plants

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German chamomile (M. chamomilla) is recognized as a star herb due to its medicinal and aromatic properties. This plant is found across a wide range of climatic and soil conditions. Both the flower heads and blue essential oils of German chamomile possess several pharmacological properties of an anti-inflammatory, antimicrobial, antiseptic, antispasmodic and sedative, etc., nature, which makes it a highly sought after herb for use in many pharma and aroma industries. Chamomile tea, prepared from its flower heads, is also a well-known herbal tea for mind and body relaxation. Though it is a high-demand herb, farmers have not adopted this plant for large scale cultivation as a crop, which could improve their livelihood, due to the high cost in flower heads harvesting, loss in over mature and immature flower heads picking during harvesting, unavailability of varieties and agrotechnologies for machine harvesting, a lack of efficient process development of oil extraction and in the lack of improved stable varieties. There are many studies that have reported on the phytochemistry and pharmacological uses of chamomile, which further explore its importance in the medicine industry. Several studies are also present in the literature on its cultivation practices and plant ecology. However, studies on breeding behavior, genetic improvement, varietal development and mechanical harvesting are scarce in German chamomile. Hence, keeping in mind various aspects of farmers’ and researchers’ interest, earlier reports on taxonomy, floral biology, processing of oil extraction, active constituents, uses, agronomy, breeding challenges and opportunities in German chamomile are summarized in this review.

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“…In nuclear transformation, one nuclear transformation vector can be applied to multiple plants. For example, a nuclear transformation vector pCHS [carrying the expression cassette for the selection marker neomycin phosphotransferase II (nptII) driven by nopaline synthase (nos) promoter and Petunia chalcone synthase (chs) driven by cauliflower mosaic virus 35S promoter] and an Agrobacterium tumefaciensmediated method were employed to a range of plant species including the highvalue medicinal plants Echinacea purpurea [11] and E. pallida [12], model plant tobacco [13], medicinal herb Bidens pilosa [14], and the floricultural plant Cleome spinosa [15]; the transfer DNA (T-DNA) containing the expression cassette in vector pCHS was integrated randomly in the nuclear chromosome of transgenic plants, and the foreign Petunia chs and selection marker nptII were detected and expressed. However, in plastid transformation, a plastid transformation vector has to be constructed for each target plant which is quite a laborious work.…”

Section: Introductionmentioning

confidence: 99%

Construction and Evaluation of Chloroplast Expression Vectors in Higher Plants

Chen¹,

Tsai²,

To³

2020

Genetic Transformation in Crops

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Plastid transformation has a number of advantages in comparison with nuclear transformation. Currently, only tobacco (Nicotiana tabacum) is routinely used in plastid transformation. Here we constructed a series of chloroplast expression vectors specific for spinach (pCEV1), tomato (pCEV2 and pCEV3), and N. benthamiana (pCEV4). Selection marker aminoglycoside 3 0-adenyltransferase (aadA) conferring spectinomycin resistance was used in pCEV1, pCEV2, and pCEV4, while selection marker neomycin phosphotransferase II (nptII) was used in pCEV3. The expression cassette in these vectors was integrated in the intergenic spacer between trnI and trnA of plastid genome via homologous recombination. Several transgenes, including a reporter gene encoding GFP:GUS fusion protein and genes from tomato (lycopene b-cyclase, z-carotene desaturase) and bamboo mosaic virus satellite RNA (encoding coat protein CP20), were independently cloned into some of these vectors. Transient GUS expression was detected in spinach leaves bombarded by pCEV1/GFP-GUS. Functional expression of selection markers aadA and nptII was demonstrated for spinach, tomato, and N. benthamiana. Seedling assay from T 0 self-pollinated plant of transplastomic N. benthamiana confirmed maternal inheritance of transgenes, and genomic PCR analysis confirmed integration of transgenic expression cassette into the plastid genome of N. benthamiana. Moreover, auxiliary vectors pECaad and pECnpt are also reported.

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In vitro regeneration, Agrobacterium-mediated transformation, and genetic assay of chalcone synthase in the medicinal plant Echinacea pallida

Cited by 7 publications

References 61 publications

Transformation and Characterization of Δ12-Fatty Acid Acetylenase and Δ12-Oleate Desaturase Potentially Involved in the Polyacetylene Biosynthetic Pathway from Bidens pilosa

Transformation and Characterization of Δ12-Fatty Acid Acetylenase and Δ12-Oleate Desaturase Potentially Involved in the Polyacetylene Biosynthetic Pathway from Bidens pilosa

A Comprehensive Review on Biology, Genetic Improvement, Agro and Process Technology of German Chamomile (Matricaria chamomilla L.)

Construction and Evaluation of Chloroplast Expression Vectors in Higher Plants

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