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

• 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
• ISSN: 0302-8933; 1432-072X
• DOI: 10.1007/s00203-022-03206-z

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.