Establishment of an efficient Agrobacterium -mediated genetic transformation method in Macleaya cordata

Huang, Peng; Xu, Min; Xia, Liqiong; Qing, Zhixing; Tang, Qi; Liu, Wei; Zeng, Jianguo

doi:10.1016/j.scienta.2017.09.004

Cited by 12 publications

(10 citation statements)

References 44 publications

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“…Then, we put the explants on sterile filter paper to remove surface moisture and excess bacteria and incubated them in darkness on co-cultivation medium (MS solid medium, 30 g/L sucrose, 8 g/L agar, pH 5.8) at 25 °C for 3 days. In previous studies, we found that 20 mg/L acetosyringone in the co-cultivation medium was the most effective concentration for induction of transformants 26 . Therefore, we used this concentration in the co-cultivation medium.…”

Section: Methodsmentioning

confidence: 87%

“…rhizogenes infection comes from an improvement on the previous transformation system of M . cordata 26 . In this study, an A .…”

Section: Resultsmentioning

confidence: 99%

“…cordata was established using A . tumefaciens 26 . However, there are two reasons for us to construct an A .…”

Section: Discussionmentioning

confidence: 99%

“…The explants were submerged in the A . rhizogenes suspension medium and submitted to a continuous vacuum for 10 min 26 . Then, we put the explants on sterile filter paper to remove surface moisture and excess bacteria and incubated them in darkness on co-cultivation medium (MS solid medium, 30 g/L sucrose, 8 g/L agar, pH 5.8) at 25 °C for 3 days.…”

Section: Methodsmentioning

confidence: 99%

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Hairy root induction and benzylisoquinoline alkaloid production in Macleaya cordata

Huang

Xia

Liu

et al. 2018

Sci Rep

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Sanguinarine is currently widely used to replace antibiotic growth promoters in animal feeding and has demonstrated useful anticancer activity. Currently, the main source of sanguinarine is from an important medicinal plant, Macleaya cordata. To obtain a new source of sanguinarine production, we established hairy root cultures of M. cordata by co-cultivating leaf and stem explants with Agrobacterium rhizogenes. Except the co-cultivation medium, all growth media contained 200 mg/L timentin to eliminate A. rhizogenes. Through comparing the metabolic profiles and gene expression of hairy roots and wild-type roots sampled at five time points, we found that the sanguinarine and dihydrosanguinarine contents of hairy roots were far higher than those of wild-type roots, and we revealed the molecular mechanism that causes these metabolites to increase. Consequently, this study demonstrated that the hairy root system has further potential for bioengineering and sustainable production of sanguinarine on a commercial scale. To the best of our knowledge, this is the first efficient protocol reported for the establishment of hairy root cultures in M. cordata using A. rhizogenes.

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Section: Methodsmentioning

confidence: 87%

“…rhizogenes infection comes from an improvement on the previous transformation system of M . cordata 26 . In this study, an A .…”

Section: Resultsmentioning

confidence: 99%

“…cordata was established using A . tumefaciens 26 . However, there are two reasons for us to construct an A .…”

Section: Discussionmentioning

confidence: 99%

Section: Methodsmentioning

confidence: 99%

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Hairy root induction and benzylisoquinoline alkaloid production in Macleaya cordata

Huang

Xia

Liu

et al. 2018

Sci Rep

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“…Recently, the whole genome of M. cordata has been de novo sequenced and validated for SAN biosynthesis 9 . In addition, a transformation and regeneration system to produce transgenic M. cordata has been constructed 23 . These genomics data and transgenic methods provide the opportunity for enhanced production of BIAs through plant metabolic engineering.…”

Section: Introductionmentioning

confidence: 99%

Modulation of benzylisoquinoline alkaloid biosynthesis by overexpression berberine bridge enzyme in Macleaya cordata

Huang

Liu

et al. 2018

Sci Rep

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Macleaya cordata produces a variety of benzylisoquinoline alkaloids (BIAs), such as sanguinarine, protopine, and berberine, which are potential anticancer drugs and natural growth promoters. The genes encoding the berberine bridge enzyme (BBE) were isolated from M. cordata and Papaver somniferum, and then the two genes were overexpressed in M. cordata. Through liquid chromatography with triple-quadrupole mass spectrometry analysis, it was determined that McBBE-OX caused higher levels of (S)-norcoclaurine, (S)-coclaurine, (S)-N-cis-methylcoclaurine, (S)-reticuline, (S)-tetrahydrocolumbamine, (S)-tetrahydroberberine, (S)-cheilanthifoline, and (S)-scoulerine than PsBBE-OX, empty vector or control treatments. qRT-PCR analysis demonstrated that the introduced genes in the transgenic lines were all highly expressed. However, the levels of sanguinarine (SAN) and chelerythrine (CHE) in all the transgenic lines were slightly lower than those in the wild-type lines, possibly because the overexpression of McBBE causes feedback-inhibition. This is the first report on the overexpression of potential key genes in M. cordata, and the findings are important for the design of metabolic engineering strategies that target BIAs biosynthesis.

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Application of genetics and biotechnology for improving medicinal plants

Niazian

2019

Planta

138

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Establishment of an efficient Agrobacterium -mediated genetic transformation method in Macleaya cordata

Cited by 12 publications

References 44 publications

Hairy root induction and benzylisoquinoline alkaloid production in Macleaya cordata

Hairy root induction and benzylisoquinoline alkaloid production in Macleaya cordata

Modulation of benzylisoquinoline alkaloid biosynthesis by overexpression berberine bridge enzyme in Macleaya cordata

Application of genetics and biotechnology for improving medicinal plants

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