Selection and validation of reference genes for normalizing qRTâPCR gene expression studies in Colletotrichum gloeosporioides and interaction with the guava plants.
Front Plant Sci
; 14: 1235848, 2023.
Article
em En
| MEDLINE
| ID: mdl-38089797
ABSTRACT
Quantitative real-time PCR is used to quantify gene expression, even to detect low-level transcripts. It detects and quantifies the inoculum level of fungal pathogens in infected hosts. However, reliable expression profiling data require accurate transcript normalization against a stable reference gene. Hence, using stably expressed reference genes under variable conditions is paramount in gene expression analysis. In the current study, reference genes were selected and validated in Colletotrichum gloeosporioides, a guava canker and dieback pathogen. The reference gene selection and validation in C. gloeosporioides were evaluated for germinated conidia and mycelium (in vitro) and in infected guava (Psidium guajava) (interaction with host plant). The CgCAL gene was determined as a highly stable reference gene, followed by the CgTUB2 in C. gloeosporioides for germinating conidia and mycelium. However, the CgTUB2 gene was determined to be a highly stable reference gene, followed by the CgCAL for expression analysis during its interaction with the plant. Expression profiling revealed stable and constant relative expression patterns of selected reference genes for both PR genes by determining their relative transcript level. This study is the first to describe reference gene selection and validation to quantify target gene expression in C. gloeosporioides.
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Base de dados:
MEDLINE
Idioma:
En
Revista:
Front Plant Sci
Ano de publicação:
2023
Tipo de documento:
Article
País de afiliação:
Paquistão