Evaluation of reference genes for normalization of mRNA and microRNA expression by RT-qPCR under different experimental conditions in Medicago ruthenica (L.) Ledeb.

Guo, Mao-Wei; Zhu, Lin; Li, Hongyan; Wu, Zinian; Li, Zhiyong; Li, Jun

doi:10.1007/s10722-021-01243-z

Cited by 3 publications

(2 citation statements)

References 41 publications

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“…Therefore, miRNA can be used as an internal reference gene and are sometimes more stable than the commonly used internal reference genes (Michael and Bowman, 2008). Recently, studies have been performed to screen for miRNAs in many species such as soybean (Kulcheski et al, 2010), wheat , watermelon (Kong et al, 2014), Chinese Cedar , sugarcane (Nandakumar et al, 2021), Medicago ruthenica (Guo et al, 2022). miR167 and miR159 were used as the internal reference miRNAs under abiotic and biotic stress in wheat .…”

Section: Expression Analysis Of Differential Mirnas After Stressmentioning

confidence: 99%

Selection of suitable reference genes for miRNA expression normalization by quantitative RTPCR under abiotic stress in Larix olgensis

Zhang,

Yan,

Yan

et al. 2023

Silvae Genetica

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Suitable selection of endogenous controls is necessity for obtaining reliable qRT-PCR results. However, insufficient attention has been paid to the selection of endogenous controls for normalization in different stresses in plant, especially Larix olgensis. In this study, we screened candidate internal reference genes for miRNA expression normalization after subjecting Larix olgensis PEG-induced drought, NaHCO3, or NaCl stress. Needles samples were collected at 0, 12, 24, 48 and 96 h after treatment. Four common housekeeping genes and 11 mature miRNAs that were found to be stably expressed in small RNA sequencing data of Larix olgensis were selected as candidate internal reference genes. Their expression levels were assessed by qRT-PCR. GeNorm and NormFinder were used to analyse the stability of gene expression from the candidate internal reference genes. Finally, the expression of some miRNAs was analysed after stress. The results showed that miRNAs exhibited better expression stability compared to the analysed ncRNAs. Under PEG, NaHCO3, or NaCl stress, miR83321, miR43169, and miR74940 were identified as the most suitable internal reference genes, respectively. Under PEG stress, the expression of most miRNAs reached its highest level at 12 h, showing a trend of first increasing expression and then decreasing. Similarly, under NaHCO3 and NaCl stress, the expression of most miRNAs reached its maximum levels at 48 h and 12 h, respectively. This indicates that most miRNAs more quickly respond to PEG and NaCl stress than NaHCO3 stress.

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Section: Expression Analysis Of Differential Mirnas After Stressmentioning

confidence: 99%

Selection of suitable reference genes for miRNA expression normalization by quantitative RTPCR under abiotic stress in Larix olgensis

Zhang,

Yan,

Yan

et al. 2023

Silvae Genetica

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“…Previous studies on drought tolerance in M. ruthenica (L.) have mostly focused on morphology, physiology, and genomics [7][8][9][10]. However, there are many problems in improving drought tolerance in lentils: traditional breeding methods are time-consuming and labor-intensive, and mutations brought about by mutation breeding are full of uncertainties.…”

Section: Introductionmentioning

confidence: 99%

Variety-Driven Effect of Rhizosphere Microbial-Specific Recruitment on Drought Tolerance of Medicago ruthenica (L.)

Xing,

Fan,

Wang

et al. 2023

Microorganisms

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As one of the environmental factors that seriously affect plant growth and crop production, drought requires an efficient but environmentally neutral approach to mitigate its harm to plants. Soil microbiomes can interact with plants and soil to improve the adverse effects of drought. Medicago ruthenica (L.) is an excellent legume forage with strong drought tolerance, but the key role of microbes in fighting drought stress remains unclear. What kind of flora plays a key role? Is the recruitment of such flora related to its genotype? Therefore, we selected three varieties of M. ruthenica (L.) for drought treatment, analyzed their growth and development as well as their physiological and biochemical characteristics, and performed 16S rRNA high-throughput sequencing analysis on their rhizosphere soils to clarify the variety-mediated response of rhizosphere bacteria to drought stress. It was found that among the three varieties of M. ruthenica (L.), Mengnong No.2, Mengnong No.1 and Zhilixing were subjected to drought stress and showed a reduction in plant height increment of 24.86%, 34.37%, and 31.97% and in fresh weight of 39.19%, 50.22%, and 41.12%, respectively, whereas dry weight was reduced by 23.26%, 26.10%, and 24.49%, respectively. At the same time, we found that the rhizosphere microbial community of Mengnong No. 2 was also less affected by drought, and it was able to maintain the diversity of rhizosphere soil microflora stable after drought stress, while Mennong No. 1 and Zhilixing were affected by drought stress, resulting in a decrease in rhizosphere soil bacterial community diversity indices to 92.92% and 82.27%, respectively. Moreover, the rhizosphere of Mengnon No. 2 was enriched with more nitrogen-fixing bacteria Rhizobium than the other two varieties of M. ruthenica (L.), which made it still have a good ability to accumulate aboveground biomass after drought stress. In conclusion, this study proves that the enrichment process of bacteria is closely related to plant genotype, and different varieties enrich different types of bacteria in the rhizosphere to help them adapt to drought stress, and the respective effects are quite different. Our results provide new evidence for the study of bacteria to improve the tolerance of plants to drought stress and lay a foundation for the screening and study mechanism of drought-tolerant bacteria in the future.

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Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress

Jing¹,

Zhou²,

Gong³

et al. 2023

J. Ocean Univ. China

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Evaluation of reference genes for normalization of mRNA and microRNA expression by RT-qPCR under different experimental conditions in Medicago ruthenica (L.) Ledeb.

Cited by 3 publications

References 41 publications

Selection of suitable reference genes for miRNA expression normalization by quantitative RTPCR under abiotic stress in Larix olgensis

Selection of suitable reference genes for miRNA expression normalization by quantitative RTPCR under abiotic stress in Larix olgensis

Variety-Driven Effect of Rhizosphere Microbial-Specific Recruitment on Drought Tolerance of Medicago ruthenica (L.)

Evaluation of Reference Genes for Quantitative Real-Time PCR Analysis in Manila Clam Ruditapes philippinarum Under Hypoxic Stress

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