Biodegradation of γ-hexachlorocyclohexane by transgenic hairy root cultures of Cucurbita moschata that accumulate recombinant bacterial LinA.
γ-HCH was successfully degraded using LinA-expressed transgenic hairy root cultures of Cucurbita moschata . Fusing an endoplasmic reticulum-targeting signal peptide to LinA was essential for stable accumulation in the hairy roots. The pesticide γ-hexachlorocyclohexane (γ-HCH) is a persistent organic pollutant (POP) that raises public health and environmental pollution concerns worldwide. Although several isolates of γ-HCH-degrading bacteria are available, inoculating them directly into γ-HCH-contaminated soil is ineffective because of the bacterial survival rate. Cucurbita species incorporate significant amounts of POPs from soils compared with other plant species. Here, we describe a novel bioremediation strategy that combines the bacterial degradation of γ-HCH and the efficient uptake of γ-HCH by Cucurbita species. We produced transgenic hairy root cultures of Cucurbita moschata that expressed recombinant bacterial linA, isolated from the bacterium Sphingobium japonicum UT26. The LinA protein was accumulated stably in the hairy root cultures by fusing an endoplasmic reticulum (ER)-targeting signal peptide to LinA. Then, we demonstrated that the cultures degraded more than 90 % of γ-HCH (1 ppm) overnight and produced the γ-HCH metabolite 1,2,4-trichlorobenzene, indicating that LinA degraded γ-HCH. These results indicate that the gene linA has high potential for phytoremediation of environmental γ-HCH.
Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan. nanasato@affrc.go.jp.|Forest Bio-Research Center, Forestry and Forest Products Research Institute, 3809-1 Ishi, Juo, Hitachi, Ibaraki, 319-1301, Japan. nanasato@affrc.go.jp.
Yoshihiko
Y
Nanasato
Graduate School of Life and Environmental Sciences, University of Tsukuba, 1-1-1 Tennodai, Tsukuba, Ibaraki, 305-8577, Japan.|Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
Sayuri
S
Namiki
Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.
Masao
M
Oshima
Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577, Japan.|The United Graduate School of Agricultural Science, Gifu University, 1-1 Yanagido, Gifu, Gifu, 501-1193, Japan.
Ryota
R
Moriuchi
Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan.|Forest Bio-Research Center, Forestry and Forest Products Research Institute, 3809-1 Ishi, Juo, Hitachi, Ibaraki, 319-1301, Japan.
Ken-Ichi
K
Konagaya
Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
Nobuyasu
N
Seike
Organochemicals Division, National Institute for Agro-Environmental Sciences, 3-1-3 Kannondai, Tsukuba, Ibaraki, 305-8604, Japan.
Takashi
T
Otani
Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577, Japan.
Yuji
Y
Nagata
Department of Environmental Life Sciences, Graduate School of Life Sciences, Tohoku University, 2-1-1 Katahira, Aoba, Sendai, Miyagi, 980-8577, Japan.
Masataka
M
Tsuda
Genetically Modified Organism Research Center, National Institute of Agrobiological Sciences, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan. tabei@affrc.go.jp.
Yutaka
Y
Tabei
D001426:Bacterial Proteins / Q000378:metabolism*; D001483:Base Sequence; D001673:Biodegradation, Environmental; D028464:Cucurbita / Q000235:genetics* / Q000378:metabolism*; D004721:Endoplasmic Reticulum / Q000378:metabolism; D001556:Hexachlorocyclohexane / Q000378:metabolism*; D018517:Plant Roots / Q000254:growth & development*; D030821:Plants, Genetically Modified; D021382:Protein Sorting Signals; D011995:Recombination, Genetic / Q000235:genetics; D016415:Sequence Alignment; D045249:Sphingobacterium / Q000378:metabolism*; D013347:Subcellular Fractions / Q000378:metabolism; D046509:Tissue Culture Techniques / Q000379:methods*
D001426:Bacterial Proteins; D021382:Protein Sorting Signals; D001556:Hexachlorocyclohexane
Biodegradation; Cucurbita moschata; Hairy root cultures; LinA; Phytoremediation; γ-Hexachlorocyclohexane (γ-HCH)