Selection and validation of reference genes for normalizing qRT‒PCR gene expression studies in Colletotrichum gloeosporioides and interaction with the guava plants

• Published in: Frontiers in plant science Vol. 14; p. 1235848
• Main Authors: Haq, Imran Ul, Ijaz, Siddra, Hashem, Abeer, Avila-Quezada, Graciela Dolores, Abd Allah, Elsayed Fathi
• Format: Journal Article
• Language: English
• Published: Switzerland Frontiers Media S.A 23-11-2023
• Subjects: Colletotrichum gloeosporioides; pathogenesis-related genes; Plant Science; Psidium guajava; qRT−PCR; reference genes; reference genes; qRT−PCR; Colletotrichum gloeosporioides; Psidium guajava; pathogenesis-related genes
• ISSN: 1664-462X; 1664-462X
• DOI: 10.3389/fpls.2023.1235848

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
, a guava canker and dieback pathogen. The reference gene selection and validation in
were evaluated for germinated conidia and mycelium (
) and in infected guava (
) (interaction with host plant). The
gene was determined as a highly stable reference gene, followed by the
in
for germinating conidia and mycelium. However, the
gene was determined to be a highly stable reference gene, followed by the
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
.