title: An efficient protocol for genetic transformation of watercress (Nasturtium officinale) using Agrobacterium rhizogenes.
abstract: Watercress (Nasturtium officinale) is a member of the Brassicaceae family and a rich source of glucosinolate, which has been shown to possess anticancer properties. To extract these compounds from N. officinale for study, a method was developed in which Agrobacterium rhizogenes was used to transfer DNA segments into plant genomes in order to produce hairy root cultures, which are a reliable source of plant compounds. The A. rhizogenes strain R1000 had the highest infection frequency and induces the most hairy roots per explant. Polymerase chain reaction and cytohistochemical staining methods were used to validate transgenic hairy roots from N. officinale. Glucosinolate from watercress hairy roots was separated and analyzed using high-performance liquid chromatography coupled to electrospray ionization mass spectrometry. Indolic glucosinolates, including glucobrassicin (0.01-0.02 μmol/g of DW) and 4-methoxyglucobrassicin (0.06-0.18 μmol/g of DW), as well as aromatic glucosinolate (gluconasturtiin) (0.06-0.21 μmol/g of DW), were identified virtually identical or more in transformed than wild type roots of N. officinale. Hairy root culture of watercress is a valuable approach for future efforts in the metabolic engineering of glucosinolate biofortification in plants, particularly, because indolic glucosinolates are the precursors of a potent cancer chemopreventive agent (indole-3-carbinol).
authors.affiliation: Department of Crop Science, College of Agriculture and Life Sciences, Chungnam National University, 220 Gung-dong, Yuseong-gu, Daejeon 305-764, Korea.
authors.forename: Nam Il
authors.initials: NI
authors.lastname: Park
authors.forename: Jae Kwang
authors.initials: JK
authors.lastname: Kim
authors.forename: Woo Tae
authors.initials: WT
authors.lastname: Park
authors.forename: Jin Woong
authors.initials: JW
authors.lastname: Cho
authors.forename: Yong Pyo
authors.initials: YP
authors.lastname: Lim
authors.forename: Sang Un
authors.initials: SU
authors.lastname: Park
mesh_terms: D060054:Agrobacterium / Q000378:metabolism*; D002851:Chromatography, High Pressure Liquid; D018548:Cotyledon / Q000235:genetics; D005821:Genetic Techniques; D005961:Glucosinolates / Q000378:metabolism; D005966:Glucuronidase / Q000737:chemistry / Q000378:metabolism; D013058:Mass Spectrometry; D031223:Nasturtium / Q000235:genetics* / Q000382:microbiology*; D018515:Plant Leaves / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000166:cytology / Q000235:genetics / Q000378:metabolism; D030821:Plants, Genetically Modified; D014170:Transformation, Genetic
chemical_list: D005961:Glucosinolates; D005966:Glucuronidase