IAgrobacterium tumefaciens/I-Mediated Transformation of the Aquatic Fungus IPhialemonium inflatum/I FBCC-F1546
Phialemonium inflatum is a useful fungus known for its ability to mineralise lignin during primary metabolism and decompose polycyclic aromatic hydrocarbons (PAHs). However, no functional genetic analysis techniques have been developed yet for this fungus, specifically in terms of transformation. In...
Saved in:
Published in | Journal of fungi (Basel) Vol. 9; no. 12 |
---|---|
Main Authors | , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
MDPI AG
01.12.2023
|
Subjects | |
Online Access | Get full text |
Cover
Loading…
Abstract | Phialemonium inflatum is a useful fungus known for its ability to mineralise lignin during primary metabolism and decompose polycyclic aromatic hydrocarbons (PAHs). However, no functional genetic analysis techniques have been developed yet for this fungus, specifically in terms of transformation. In this study, we applied an Agrobacterium tumefaciens-mediated transformation (ATMT) system to P. inflatum for a functional gene analysis. We generated 3689 transformants using the binary vector pSK1044, which carried either the hygromycin B phosphotransferase (hph) gene or the enhanced green fluorescent protein (eGFP) gene to label the transformants. A Southern blot analysis showed that the probability of a single copy of T-DNA insertion was approximately 50% when the co-cultivation of fungal spores and Agrobacterium tumefaciens cells was performed at 24–36 h, whereas at 48 h, it was approximately 35.5%. Therefore, when performing gene knockout using the ATMT system, the co-cultivation time was reduced to ≤36 h. The resulting transformants were mitotically stable, and a PCR analysis confirmed the genes’ integration into the transformant genome. Additionally, hph and eGFP gene expressions were confirmed via PCR amplification and fluorescence microscopy. This optimised transformation system will enable functional gene analyses to study genes of interest in P. inflatum. |
---|---|
AbstractList | Phialemonium inflatum is a useful fungus known for its ability to mineralise lignin during primary metabolism and decompose polycyclic aromatic hydrocarbons (PAHs). However, no functional genetic analysis techniques have been developed yet for this fungus, specifically in terms of transformation. In this study, we applied an Agrobacterium tumefaciens-mediated transformation (ATMT) system to P. inflatum for a functional gene analysis. We generated 3689 transformants using the binary vector pSK1044, which carried either the hygromycin B phosphotransferase (hph) gene or the enhanced green fluorescent protein (eGFP) gene to label the transformants. A Southern blot analysis showed that the probability of a single copy of T-DNA insertion was approximately 50% when the co-cultivation of fungal spores and Agrobacterium tumefaciens cells was performed at 24–36 h, whereas at 48 h, it was approximately 35.5%. Therefore, when performing gene knockout using the ATMT system, the co-cultivation time was reduced to ≤36 h. The resulting transformants were mitotically stable, and a PCR analysis confirmed the genes’ integration into the transformant genome. Additionally, hph and eGFP gene expressions were confirmed via PCR amplification and fluorescence microscopy. This optimised transformation system will enable functional gene analyses to study genes of interest in P. inflatum. |
Audience | Academic |
Author | Kim, Youngjun Yoon, Jonghan Jeong, Min-Hye Jo, Miju An, Sunmin Jang, Seol-Hwa Jeong, Haejun Park, Jiwon Park, Jiyoon Goh, Jaeduk Park, Sangkyu Park, Sook-Young Kim, Seoyeon |
Author_xml | – sequence: 1 fullname: Yoon, Jonghan – sequence: 2 fullname: Kim, Youngjun – sequence: 3 fullname: Kim, Seoyeon – sequence: 4 fullname: Jeong, Haejun – sequence: 5 fullname: Park, Jiyoon – sequence: 6 fullname: Jeong, Min-Hye – sequence: 7 fullname: Park, Sangkyu – sequence: 8 fullname: Jo, Miju – sequence: 9 fullname: An, Sunmin – sequence: 10 fullname: Park, Jiwon – sequence: 11 fullname: Jang, Seol-Hwa – sequence: 12 fullname: Goh, Jaeduk – sequence: 13 fullname: Park, Sook-Young |
BookMark | eNqVj81qwzAQhEVJoWmbS59AL-BYshLbOrqmpj4Eesiht6LKK0fBkqh-3j8K9JBr2MPsDPsN7DNaWWcBoTdKtoxxUp6d4rSidN8-oHXFCC9q0n6vbvYntAnhTAjJNzXnbI3s2M3e_QoZwetkcEwGlJAabCjH4gCTFhEmfPTCBuW8EVE7i53C8QS4-0vZSzwkO6eAx6-TFgsYZ69N2qpF5LpyxMN73xcD3e_qV_SoxBJg868vaDt8HPvPYs7gT0Zc9ELmmcBomd9TOudd03Dats2uYncDF4vyWWk |
ContentType | Journal Article |
Copyright | COPYRIGHT 2023 MDPI AG |
Copyright_xml | – notice: COPYRIGHT 2023 MDPI AG |
DOI | 10.3390/jof9121158 |
DatabaseTitleList | |
DeliveryMethod | fulltext_linktorsrc |
Discipline | Botany |
EISSN | 2309-608X |
ExternalDocumentID | A779188742 |
GeographicLocations | South Korea Japan |
GeographicLocations_xml | – name: South Korea – name: Japan |
GroupedDBID | 53G 5VS 8FE 8FH AADQD AAFWJ ABDBF ADBBV AFKRA AFPKN AFZYC ALMA_UNASSIGNED_HOLDINGS AOIJS BBNVY BCNDV BENPR BHPHI CCPQU GROUPED_DOAJ HCIFZ HYE IAO KQ8 LK8 M7P MODMG M~E OK1 PIMPY PROAC RPM |
ID | FETCH-gale_infotracacademiconefile_A7791887423 |
ISSN | 2309-608X |
IngestDate | Tue Jan 23 06:11:57 EST 2024 |
IsPeerReviewed | true |
IsScholarly | true |
Issue | 12 |
Language | English |
LinkModel | OpenURL |
MergedId | FETCHMERGED-gale_infotracacademiconefile_A7791887423 |
ParticipantIDs | gale_infotracacademiconefile_A779188742 |
PublicationCentury | 2000 |
PublicationDate | 20231201 |
PublicationDateYYYYMMDD | 2023-12-01 |
PublicationDate_xml | – month: 12 year: 2023 text: 20231201 day: 01 |
PublicationDecade | 2020 |
PublicationTitle | Journal of fungi (Basel) |
PublicationYear | 2023 |
Publisher | MDPI AG |
Publisher_xml | – name: MDPI AG |
SSID | ssj0001586993 |
Score | 4.5882626 |
Snippet | Phialemonium inflatum is a useful fungus known for its ability to mineralise lignin during primary metabolism and decompose polycyclic aromatic hydrocarbons... |
SourceID | gale |
SourceType | Aggregation Database |
SubjectTerms | DNA Ethylenediaminetetraacetic acid Fluorescence Fluorescence microscopy Fungi Gene expression Genes Genomics Lignin Physiological aspects Polycyclic aromatic hydrocarbons |
Title | IAgrobacterium tumefaciens/I-Mediated Transformation of the Aquatic Fungus IPhialemonium inflatum/I FBCC-F1546 |
Volume | 9 |
hasFullText | 1 |
inHoldings | 1 |
isFullTextHit | |
isPrint | |
link | http://utb.summon.serialssolutions.com/2.0.0/link/0/eLvHCXMwnV3da8IwEA9D97CXsU_2KXkY7EGqs_Yrj62us4JDmAP3JLGmOnEtc-2D--t3Saup4sDtpZQjHEful7sfSe6C0B0kBUO1qK4QRlRFMwJdocw3lYCOGNMos4whr3fuPButV63d1_vyBF9Ul8TDiv-9ta7kP14FGfiVV8n-wbMrpSCAf_AvfMHD8N3Jx549nsN6FP2Wk49yDHEmoHyxgnNcT-mIZziAUfZy7DTlh6Kg5DMR7VpdWO_JV9nrTsBiBsZzXWDejMb8EWTXK7tOo6G4QIOMX7gsZMfxOyerDiTFWW534S1KT_XbUTie0NyJv4ChCDXTZFP8wqIFk9cD2iy7NtyibDk426dQ6xt3PjrNrle2n2R4U_nZjvEgHhaGTLRFlsVnkoehui3s1-uE35OcRgHhHevSXvAbbbRt0yQ1iKga5OuiahJdL6Ci7TQdV27J6ZZBRJvmlR1pQ1uuvyq1Z9k7x0N6R-gwm3Rsp2g4RnssPEH7TgQkf3GKwnVI4BwkqhIQeB0QOAowAAJngMApIPAaIPASEFUPSzicoYr72Gu0FG7pgE9WPKc-zWouopDxtl8DOSn1c1QIQXyBsOlrhFEGoR-o30i3LKrqPm9dWNMYDXzrEt3vqPRq55HX6EBi5gYV4nnCboHtxcNS5qWS2C35ASlSXPs |
link.rule.ids | 315,786,790,870,27955,27956 |
linkProvider | EBSCOhost |
openUrl | ctx_ver=Z39.88-2004&ctx_enc=info%3Aofi%2Fenc%3AUTF-8&rfr_id=info%3Asid%2Fsummon.serialssolutions.com&rft_val_fmt=info%3Aofi%2Ffmt%3Akev%3Amtx%3Ajournal&rft.genre=article&rft.atitle=IAgrobacterium+tumefaciens%2FI-Mediated+Transformation+of+the+Aquatic+Fungus+IPhialemonium+inflatum%2FI+FBCC-F1546&rft.jtitle=Journal+of+fungi+%28Basel%29&rft.au=Yoon%2C+Jonghan&rft.au=Kim%2C+Youngjun&rft.au=Kim%2C+Seoyeon&rft.au=Jeong%2C+Haejun&rft.date=2023-12-01&rft.pub=MDPI+AG&rft.issn=2309-608X&rft.eissn=2309-608X&rft.volume=9&rft.issue=12&rft_id=info:doi/10.3390%2Fjof9121158&rft.externalDocID=A779188742 |
thumbnail_l | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/lc.gif&issn=2309-608X&client=summon |
thumbnail_m | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/mc.gif&issn=2309-608X&client=summon |
thumbnail_s | http://covers-cdn.summon.serialssolutions.com/index.aspx?isbn=/sc.gif&issn=2309-608X&client=summon |