Transcriptome-guided selection of reference genes from field-grown adult trees and seedlings exposed to defense phytohormones in Erythrina velutina

IF 2.2 Q3 GENETICS & HEREDITY

Plant Gene Pub Date : 2025-02-28 DOI:10.1016/j.plgene.2025.100496

Daisy Sotero Chacon , Cibele Tesser da Costa , Fábio Antônio Antonelo , Bernardo Bonilauri , Taffarel Melo Torres , Raquel Brandt Giordani , Arthur Germano Fett-Neto

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引用次数: 0

Abstract

Erythrina velutina Willd (Fabaceae) is an important non-model tree with pharmaceutical potential, traditionally used in folk medicine at the northeastern Brazilian semiarid region. The species shows anti-inflammatory properties and central nervous system modulation. Our previous work highlighted its capacity of biosynthesizing several bioactive compounds. It was also possible to identify enzyme and transcription factor genes involved in the response to both biotic and abiotic stresses. However, the identification of reference genes (RGs) for real-time quantitative polymerase chain reaction (RT-qPCR) assays and validation of molecular mechanisms is still lacking. Leaves and seeds of field-grown plants and shoots of seedlings exposed to defense phytohormones (sodium nitroprusside and methyl jasmonate, the former as source of nitric oxide) were examined. As a result, 60 transcripts had RPKM (log2) > 2 and CV < 10 %, from which the 11 most conserved were selected: NADH, 26S, UPF, PBL27, CYB5, ARP4, ARD2, OAT, CSTF64, NLE, and RAB7. CSTF64, RAB7, NLE, UPF and ARP4 exhibited the highest stability and showed positive, and significant correlations, while NADH and PBL27 were the least stable genes. The use of two genes was sufficient for normalizing qPCR assays, and the best gene pairs were recommended. All reference genes were validated with target genes, sequenced, and showed adequate melting curves and amplification efficiency. This pioneering identification of reference genes in Erythrina velutina provides a platform for future molecular studies in this unique dryland species. The herein described approach can also be useful in identifying adequate housekeeping genes in other non-model plant species.

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毛毛Erythrina velutina田间成树和幼苗防御激素内参基因的转录组引导选择

Erythrina velutina Willd（豆科）是一种重要的非模式树种，具有制药潜力，传统上用于巴西东北部半干旱地区的民间医药。该物种具有抗炎和调节中枢神经系统的特性。我们之前的工作强调了它生物合成多种生物活性化合物的能力。此外，我们还发现了参与应对生物和非生物压力的酶和转录因子基因。然而，目前仍缺乏用于实时定量聚合酶链反应（RT-qPCR）测定和分子机制验证的参考基因（RGs）的鉴定。研究人员对田间生长植物的叶片和种子以及暴露于防御植物激素（硝普钠和茉莉酸甲酯，前者是一氧化氮的来源）的幼苗嫩枝进行了检测。结果发现，60 个转录本的 RPKM (log2) > 2 和 CV < 10 %，从中选出了 11 个最保守的转录本：NADH、26S、UPF、PBL27、CYB5、ARP4、ARD2、OAT、CSTF64、NLE 和 RAB7。CSTF64、RAB7、NLE、UPF 和 ARP4 表现出最高的稳定性和显著的正相关性，而 NADH 和 PBL27 是最不稳定的基因。使用两个基因就足以对 qPCR 检测进行归一化，推荐使用最佳基因对。所有参考基因都与目标基因进行了验证、测序，并显示出足够的熔解曲线和扩增效率。这次对 Erythrina velutina 参考基因的开创性鉴定为今后对这一独特旱地物种进行分子研究提供了一个平台。本文所述方法也可用于鉴定其他非模式植物物种的适当管家基因。

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来源期刊

Plant Gene Agricultural and Biological Sciences-Plant Science

CiteScore

4.50

自引率

0.00%

发文量

审稿时长

51 days

期刊介绍： Plant Gene publishes papers that focus on the regulation, expression, function and evolution of genes in plants, algae and other photosynthesizing organisms (e.g., cyanobacteria), and plant-associated microorganisms. Plant Gene strives to be a diverse plant journal and topics in multiple fields will be considered for publication. Although not limited to the following, some general topics include: Gene discovery and characterization, Gene regulation in response to environmental stress (e.g., salinity, drought, etc.), Genetic effects of transposable elements, Genetic control of secondary metabolic pathways and metabolic enzymes. Herbal Medicine - regulation and medicinal properties of plant products, Plant hormonal signaling, Plant evolutionary genetics, molecular evolution, population genetics, and phylogenetics, Profiling of plant gene expression and genetic variation, Plant-microbe interactions (e.g., influence of endophytes on gene expression; horizontal gene transfer studies; etc.), Agricultural genetics - biotechnology and crop improvement.