Selection and Validation of Reference Genes in Different Tissues of Okra (Abelmoschus esculentus L.) under Different Abiotic Stresses

Abstract

Okra (Abelmoschus esculentus L.) is a particular vegetable with both edible and medicinal values. However, the expression pattern of the okra reference genes in response to abiotic stress has not been explored. In the present study, 18 potential reference genes were selected from okra in various tissues and abiotic stress conditions, and their expression levels were detected by Real-Time quantitative PCR (RT-qPCR). Their expression stabilities were calculated by four algorithms (geNorm, NormFinder, BestKeeper, and RefFinder). Under cold stress, the most stable genes included GAPC1 and CYP (leaf), CYP and ACT7 (root), HIS6 and GAPC1 (stem), and HIS6 and 60s (different tissues). Under salt stress, EF-1α and UBQ (leaf), EF-1α and UBQ (root), TUA4 and Eif (stem), and HIS6 and Eif (different tissues) were the most stable genes. Under drought stress, UBQ and Eif in the leaf, HIS6 and Eif in the root, TUA4 and HIS6 in the stem, and UBQ and Eif in different tissues were most stably expressed in okra. In addition, complete sequencing results by RefFinder showed that HIS6 and ACT7 in the leaf, HIS6 and Eif in the root, UBC5B and 60s in the stem, and HIS6 and Eif in different tissues, were most the suitable reference genes for okra. Furthermore, AeMYB1R1 transcription factor was used to verify the reliability of RT-qPCR values. In summary, this study was carried out to demonstrate the potential reference genes of okra under abiotic stress, aiming to provide a molecular basis for functional gene analysis and regulatory mechanism research of okra.

Selection of Reference Genes in Evodia rutaecarpa var. officinalis and Expression Patterns of Genes Involved in Its Limonin Biosynthesis

“…Many researchers have conducted relatively quantitative comparisons between the most stable and unstable reference genes, and they have often obtained significant differences. The use of unstable reference genes leads to inaccurate quantitative results [ 30 , 31 , 32 ]. Specifically, in the case of fruits at different developmental stages, the two most stable reference genes, Act1 and UBQ , should be employed, while Act1 and His3 are advisable choices for different tissues.…”

Appressoria Formation in Phytopathogenic Fungi Suppressed by Antimicrobial Peptides and Hybrid Peptides from Black Soldier Flies

“…Then, using RNAase-free mortars and pestles (Beijing, China), the samples were crushed, and a powder was obtained. The total RNA was extracted and purified using TRIzol [38] (Ambion, Austin, TX, USA) according to the manufacturer's instructions. The RNA quality and concentration were determined using NanoDrop (BioRad, Hercules, California, USA) after dissolving the RNAs in DEPC water, and then, 20 µL of RNA aliquot was stored at −80 • C.…”