Transient expression in electroporated pea protoplasts: Elicitor responsiveness of a phenylalanine ammonia-lyase promoter |
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Authors: | Tadaaki Hashimoto Tetsuji Yamada Akiko Tada Shinji Kawamata Yoshikazu Tanaka Pernpong Sriprasertsak Yuki Ichinose Hisaharu Kato Satoshi Izutsu Tomonori Shiraishi Hachiro Oku Yoshiaki Ohtsuki |
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Institution: | (1) Laboratory of Plant Pathology & Genetic Engineering, College of Agriculture, Okayama University, Tsushima, 700 Okayama, Japan;(2) Agricultural Research Center, Kannondai 3-1-1, 305 Tsukuba, Japan |
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Abstract: | Summary High yields of viable pea protoplasts were produced from suspension cultured cells and the conditions for the optimum transient expression of the chloramphenicol acetyltransferase (CAT) gene fused to the CaMV 35S promoter after electroporation were investigated. Conditions for elicitor induction of a member of the phenylalanine ammonia-lyase (PAL) gene family in pea was also investigated using a chimeric gene carrying 480 bp of the putative promoter region of gPAL1 connected to bacterial cat gene and nos terminator. CAT activity was considerably induced by the treatment with fungal elicitor (>100 g/ml glucose equivalent) isolated from Mycosphaerella pinodes, a pea pathogen.Abbreviations CAT
chloramphenicol acetyltransferase
- PAL
phenylalanine ammonia-lyase
- CM
acetylated chloramphenicol
- GSH
reduced glutathione
- NOS
nopaline synthase
- ES
electroporation solution
- CaMV
cauliflower mosaic virus
- GUS
-glucuronidase
- CHS
chalcone synthase
- 2,4-D
2, 4-dichlorophenoxyacetic acid
Present address: Research Institute, Takasago Perfumery Inc, 5-31-36, Kamata, Minato, Tokyo, 144, Japan Toso Inc, 4560 Oaza-Tomita, Shin-nanyo, Yamaguchi, 746, Japan Central Laboratory of Green Complex, Kasetsert University, Kamphaensaen, Nakohn Pathom, Thailand |
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Keywords: | Pisum sativum L Midoriusui Protoplast Suspension culture Phenylalanine ammonia-lyase Transient assay |
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