A novel gene, Le-Dd10, is involved in fruiting body formation of Lentinula edodes

The cDNA library prepared from Lentinula edodes , Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum prestalk as a probe. Twenty-one clones, Le-Dd1 ~ 21, were isolated from the L. edodes primordia cDNA library. Functional analysis of each gene w...

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Published in:	Archives of microbiology Vol. 204; no. 10; p. 602
Main Authors:	Kishikawa, Akihiro, Hamada, Satoshi, Kamei, Ichiro, Fujimoto, Yosuke, Miyazaki, Kazuhiro, Yoshida, Motonobu
Format:	Journal Article
Language:	English
Published:	Berlin/Heidelberg Springer Berlin Heidelberg 01-10-2022 Springer Nature B.V
Subjects:	Antibodies Biochemistry Biomedical and Life Sciences Biotechnology Cell Biology Cell culture Ecology Fruit bodies Functional analysis Lentinula edodes Life Sciences Microbial Ecology Microbiology Molecular weight Mushrooms Original Paper Polyclonal antibodies Polymorphism Primordia Proteins Western blotting Shiitake mushroom Transformation Transcription factor Fruiting body
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Abstract	The cDNA library prepared from Lentinula edodes , Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum prestalk as a probe. Twenty-one clones, Le-Dd1 ~ 21, were isolated from the L. edodes primordia cDNA library. Functional analysis of each gene was carried out by transformation into protoplast cells from L. edodes Mori 252 (M252) mycelia with the overexpression vector pLG-RasF1 of each gene because M252 protoplast cells were transformed with an 11-fold higher efficiency than H600 cells. Transformants with the overexpression vector of Le-Dd10 formed a fruiting body at almost the same time as H600, a positive control, although M252, a negative control, did not form a fruiting body under culture conditions. This suggested that Le-Dd10 is involved in the formation of fruiting bodies. Single-strand conformation polymorphism analysis revealed that Le-Dd10 is located on No. 4 linkage group of L. edodes . The properties of Le-Dd10 products were investigated by Western blotting analysis using polyclonal antibodies against GST:Le-Dd10 fusion proteins. As a result, 56-kDa, 27-kDa, and 14-kDa protein bands appeared in primordial and fruiting body stages, although the expected molecular weight of the Le-Dd10 product was 50 kDa.
AbstractList	The cDNA library prepared from Lentinula edodes , Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum prestalk as a probe. Twenty-one clones, Le-Dd1 ~ 21, were isolated from the L. edodes primordia cDNA library. Functional analysis of each gene was carried out by transformation into protoplast cells from L. edodes Mori 252 (M252) mycelia with the overexpression vector pLG-RasF1 of each gene because M252 protoplast cells were transformed with an 11-fold higher efficiency than H600 cells. Transformants with the overexpression vector of Le-Dd10 formed a fruiting body at almost the same time as H600, a positive control, although M252, a negative control, did not form a fruiting body under culture conditions. This suggested that Le-Dd10 is involved in the formation of fruiting bodies. Single-strand conformation polymorphism analysis revealed that Le-Dd10 is located on No. 4 linkage group of L. edodes . The properties of Le-Dd10 products were investigated by Western blotting analysis using polyclonal antibodies against GST:Le-Dd10 fusion proteins. As a result, 56-kDa, 27-kDa, and 14-kDa protein bands appeared in primordial and fruiting body stages, although the expected molecular weight of the Le-Dd10 product was 50 kDa. The cDNA library prepared from Lentinula edodes, Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum prestalk as a probe. Twenty-one clones, Le-Dd1 ~ 21, were isolated from the L. edodes primordia cDNA library. Functional analysis of each gene was carried out by transformation into protoplast cells from L. edodes Mori 252 (M252) mycelia with the overexpression vector pLG-RasF1 of each gene because M252 protoplast cells were transformed with an 11-fold higher efficiency than H600 cells. Transformants with the overexpression vector of Le-Dd10 formed a fruiting body at almost the same time as H600, a positive control, although M252, a negative control, did not form a fruiting body under culture conditions. This suggested that Le-Dd10 is involved in the formation of fruiting bodies. Single-strand conformation polymorphism analysis revealed that Le-Dd10 is located on No. 4 linkage group of L. edodes. The properties of Le-Dd10 products were investigated by Western blotting analysis using polyclonal antibodies against GST:Le-Dd10 fusion proteins. As a result, 56-kDa, 27-kDa, and 14-kDa protein bands appeared in primordial and fruiting body stages, although the expected molecular weight of the Le-Dd10 product was 50 kDa.
ArticleNumber	602
Author	Hamada, Satoshi Kamei, Ichiro Fujimoto, Yosuke Miyazaki, Kazuhiro Kishikawa, Akihiro Yoshida, Motonobu
Author_xml	– sequence: 1 givenname: Akihiro surname: Kishikawa fullname: Kishikawa, Akihiro organization: Department of Agricultural Science, Kinki University – sequence: 2 givenname: Satoshi surname: Hamada fullname: Hamada, Satoshi organization: Department of Agricultural Science, Kinki University – sequence: 3 givenname: Ichiro surname: Kamei fullname: Kamei, Ichiro organization: Department of Agricultural Science, Kinki University – sequence: 4 givenname: Yosuke surname: Fujimoto fullname: Fujimoto, Yosuke organization: Department of Agricultural Science, Kinki University – sequence: 5 givenname: Kazuhiro surname: Miyazaki fullname: Miyazaki, Kazuhiro organization: Kyushu Research Center, Forest Products Research Institute – sequence: 6 givenname: Motonobu orcidid: 0000-0002-7698-6895 surname: Yoshida fullname: Yoshida, Motonobu email: yoshida3m@gmail.com organization: Department of Agricultural Science, Kinki University, Osaka University of Comprehensive Children Education
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Keywords	Shiitake mushroom Transformation Transcription factor Fruiting body
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Snippet	The cDNA library prepared from Lentinula edodes , Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum... The cDNA library prepared from Lentinula edodes, Hokken 600 (H600), primordia was screened using cDNA expressed specifically in Dictyostelium discoideum...
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SubjectTerms	Antibodies Biochemistry Biomedical and Life Sciences Biotechnology Cell Biology Cell culture Ecology Fruit bodies Functional analysis Lentinula edodes Life Sciences Microbial Ecology Microbiology Molecular weight Mushrooms Original Paper Polyclonal antibodies Polymorphism Primordia Proteins Western blotting
Title	A novel gene, Le-Dd10, is involved in fruiting body formation of Lentinula edodes
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