Screening and Verification of Reference Genes for Analysis of Gene Expression in Garlic ( Allium sativum L.) under Cold and Drought Stress

The principal objective of this study was to screen and verify reference genes appropriate for gene expression evaluation during plant growth and development under distinct growth conditions. Nine candidate reference genes were screened based on garlic transcriptome sequence data. RT-qPCR was used t...

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Bibliographic Details
Published in:Plants (Basel) 2023-02, Vol.12 (4), p.763
Main Authors: Wang, Qizhang, Guo, Chunqian, Yang, Shipeng, Zhong, Qiwen, Tian, Jie
Format: Article
Language:English
Subjects:
Abundance
Allium sativum
Ascorbic acid
China
Cold
Cold weather
Drought
Droughts
Efficiency
Environmental aspects
Garlic
Gene expression
Genes
Genetic aspects
Glyceraldehyde-3-phosphate dehydrogenase
Growth conditions
L-Ascorbate peroxidase
normalization
Peroxidase
Physiological aspects
Plant growth
Quality standards
Rain
reference gene
RNA
rRNA 18S
RT-qPCR
Software
Stress (Physiology)
Transcriptomes
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Summary:The principal objective of this study was to screen and verify reference genes appropriate for gene expression evaluation during plant growth and development under distinct growth conditions. Nine candidate reference genes were screened based on garlic transcriptome sequence data. RT-qPCR was used to detect the expression levels of the aforementioned reference genes in specific tissues under drought and cold stress. Then, geNorm, NormFinder, BestKeeper, and ReFinder were used to consider the consistency of the expression levels of candidate reference genes. Finally, the stress-responsive gene expression of ascorbate peroxidase ( ) was quantitatively evaluated to confirm the chosen reference genes. Our results indicated that there were variations in the abundance and stability of nine reference gene transcripts underneath cold and drought stress, among which and had the highest transcript abundance, and and had the lowest transcript abundance. and were the most stably expressed genes throughout all samples; and were the most stably expressed genes during cold stress, and and were the most stably expressed genes under drought stress. The most stably expressed genes in roots, pseudostems, leaves, and cloves were , , , and , respectively, while was the most unstable gene during drought and cold stress conditions and in exclusive tissues. Taking the steady reference genes , and as references during drought and cold stress, the reliability of the expression levels was further demonstrated by detecting the expression of . Our work thereby offers a theoretical reference for the evaluation of gene expression in garlic in various tissues and under stress conditions.
ISSN:2223-7747
2223-7747
DOI:10.3390/plants12040763