Hairy Root Transformation: A Useful Tool to Explore Gene Function and Expression in Salix spp. Recalcitrant to Transformation

Gomes, Carolina; Dupas, Annabelle; Pagano, Andrea; Grima‐Pettenati, Jacqueline; Paiva, Jorge A P

doi:10.3389/fpls.2019.01427

Cited by 23 publications

(15 citation statements)

References 46 publications

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“…HR cultures have also shown promising biosynthetic ability as producers of various biologically active substances and provided insights into root metabolism [ 40 , 41 ]. Moreover, A. rhizogenes -mediated hairy root transformation systems are suitable for species recalcitrant to transformation by A. tumefaciens , as higher transformation efficiencies are obtained in comparison to A. tumefaciens -mediated transformation systems [ 42 ]. They may also help faster production of mutants using the CRISPR/CAS9 technology [ 43 ].…”

Section: Discussionmentioning

confidence: 99%

WUSCHEL Overexpression Promotes Callogenesis and Somatic Embryogenesis in Medicago truncatula Gaertn

Kadri

March

Guérineau

et al. 2021

Plants

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The induction of plant somatic embryogenesis is often a limiting step for plant multiplication and genetic manipulation in numerous crops. It depends on multiple signaling developmental processes involving phytohormones and the induction of specific genes. The WUSCHEL gene (WUS) is required for the production of plant embryogenic stem cells. To explore a different approach to induce somatic embryogenesis, we have investigated the effect of the heterologous ArabidopsisWUS gene overexpression under the control of the jasmonate responsive vsp1 promoter on the morphogenic responses of Medicago truncatula explants. WUS expression in leaf explants increased callogenesis and embryogenesis in the absence of growth regulators. Similarly, WUS expression enhanced the embryogenic potential of hairy root fragments. The WUS gene represents thus a promising tool to develop plant growth regulator-free regeneration systems or to improve regeneration and transformation efficiency in recalcitrant crops.

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Section: Discussionmentioning

confidence: 99%

WUSCHEL Overexpression Promotes Callogenesis and Somatic Embryogenesis in Medicago truncatula Gaertn

Kadri

March

Guérineau

et al. 2021

Plants

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“…Given that sweet potato is vegetatively propagated, an in vivo root transgenic system is not required for the sterilization of explants prior to A. rhizogenes infection 26,31 . The fluorescent marker DsRed used here helps to identify TRs from nontransgenic ARs of the same seedling 31 (Fig. 2b).…”

Section: Discussionmentioning

confidence: 99%

Overexpression of phosphatidylserine synthase IbPSS1 affords cellular Na+ homeostasis and salt tolerance by activating plasma membrane Na+/H+ antiport activity in sweet potato roots

Yu¹,

Xuan²,

Bian³

et al. 2020

Hortic Res

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Phosphatidylserine synthase (PSS)-mediated phosphatidylserine (PS) synthesis is crucial for plant development. However, little is known about the contribution of PSS to Na + homeostasis regulation and salt tolerance in plants. Here, we cloned the IbPSS1 gene, which encodes an ortholog of Arabidopsis AtPSS1, from sweet potato (Ipomoea batatas (L.) Lam.). The transient expression of IbPSS1 in Nicotiana benthamiana leaves increased PS abundance. We then established an efficient Agrobacterium rhizogenes-mediated in vivo root transgenic system for sweet potato. Overexpression of IbPSS1 through this system markedly decreased cellular Na + accumulation in salinized transgenic roots (TRs) compared with adventitious roots. The overexpression of IbPSS1 enhanced salt-induced Na + /H + antiport activity and increased plasma membrane (PM) Ca 2+-permeable channel sensitivity to NaCl and H 2 O 2 in the TRs. We confirmed the important role of IbPSS1 in improving salt tolerance in transgenic sweet potato lines obtained from an Agrobacterium tumefaciens-mediated transformation system. Similarly, compared with the wild-type (WT) plants, the transgenic lines presented decreased Na + accumulation, enhanced Na + exclusion, and increased PM Ca 2+-permeable channel sensitivity to NaCl and H 2 O 2 in the roots. Exogenous application of lysophosphatidylserine triggered similar shifts in Na + accumulation and Na + and Ca 2+ fluxes in the salinized roots of WT. Overall, this study provides an efficient and reliable transgenic method for functional genomic studies of sweet potato. Our results revealed that IbPSS1 contributes to the salt tolerance of sweet potato by enabling Na + homeostasis and Na + exclusion in the roots, and the latter process is possibly controlled by PS reinforcing Ca 2+ signaling in the roots.

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“…A. rhizogenes -mediated hairy root transformation systems provide an alternative to species recalcitrant to transformation by A. tumefaciens [ 17 ]. Compared to A. tumefaciens -mediated transformation, A. rhizogenes -mediated transformation system is speedy, since transgenic hairy roots grow rapidly without a complex cultured process to regenerated plantlets [ 18 , 19 ]. Therefore, A .…”

Section: Introductionmentioning

confidence: 99%

Agrobacterium rhizogenes-mediated hairy root transformation as an efficient system for gene function analysis in Litchi chinensis

Qin

Wang

et al. 2021

Plant Methods

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Background Litchi chinensis Sonn. is an economically important fruit tree in tropical and subtropical regions. However, litchi functional genomics is severely hindered due to its recalcitrance to regeneration and stable transformation. Agrobacterium rhizogenes-mediated hairy root transgenic system provide an alternative to study functional genomics in woody plants. However, the hairy root transgenic system has not been established in litchi. Results In this study, we report a rapid and highly efficient A. rhizogenes-mediated co-transformation system in L. chinensis using Green Fluorescent Protein (GFP) gene as a marker. Both leaf discs and stem segments of L. chinensis cv. ‘Fenhongguiwei’ seedlings were able to induce transgenic hairy roots. The optimal procedure involved the use of stem segments as explants, infection by A. rhizogenes strain MSU440 at optical density (OD600) of 0.7 for 10 min and co-cultivation for 3 days, with a co-transformation efficiency of 9.33%. Furthermore, the hairy root transgenic system was successfully used to validate the function of the key anthocyanin regulatory gene LcMYB1 in litchi. Over-expression of LcMYB1 produced red hairy roots, which accumulated higher contents of anthocyanins, proanthocyanins, and flavonols. Additionally, the genes involving in the flavonoid pathway were strongly activated in the red hairy roots. Conclusion We first established a rapid and efficient transformation system for the study of gene function in hairy roots of litchi using A. rhizogenes strain MSU440 by optimizing parameters. This hairy root transgenic system was effective for gene function analysis in litchi using the key anthocyanin regulator gene LcMYB1 as an example.

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Hairy Root Transformation: A Useful Tool to Explore Gene Function and Expression in Salix spp. Recalcitrant to Transformation

Cited by 23 publications

References 46 publications

WUSCHEL Overexpression Promotes Callogenesis and Somatic Embryogenesis in Medicago truncatula Gaertn

WUSCHEL Overexpression Promotes Callogenesis and Somatic Embryogenesis in Medicago truncatula Gaertn

Overexpression of phosphatidylserine synthase IbPSS1 affords cellular Na+ homeostasis and salt tolerance by activating plasma membrane Na+/H+ antiport activity in sweet potato roots

Agrobacterium rhizogenes-mediated hairy root transformation as an efficient system for gene function analysis in Litchi chinensis

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