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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 (OD 600 ) 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.

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