Stability determination of candidate reference genes in cucumber plants subjected to stresses from Phytophthora melonis
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Abstract
Stable reference genes are indispensable for ensuring the fidelity of determined gene expression levels. However, the expression levels of reference genes are unable to remain constant under all possible experimental conditions. Therefore, the stability determination of reference genes is necessary in an experimental system set. In the preset study, the stability of nine cucumber candidate reference genes (CsACT, CsUBQ, CsEF1α, CsCYP, CsαTU, CsCACS, CsTIP41, CsYSL8 and CsHEL) subjected to stresses from Phytophthora melonis(P. melonis) were determined using four different analysis methods, including Delta Ct, BestKeeper, NormFinder and GeNorm. The study results revealed that CsUBQ and CsCYP were the most stable genes suitable as internal control in cucumber plants under attack by P. melonis condition.
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Jain M, Kaur N, Garg R, Thakur JK, Tyagi AK, et al. Structure and expression analysis of early auxin-responsive Aux/IAA gene family in rice (Oryza sativa). Funct Integr Genomics. 2006; 6: 47-59. Ref.: https://tinyurl.com/y4pl23bf
Gantasala NP, Papolu PK, Thakur PK, Kamaraju D, Sreevathsa RU. Selection and validation of reference genes for quantitative gene expression studies by real-time PCR in eggplant (Solanum melongena L). BMC Res Notes. 2013; 6: 312. Ref.: https://tinyurl.com/y4xoe9cz
Thellin O, Zorzi W, Lakaye B, De-Borman B, Coumans B, et al. Husekeeping genes as internal standards: use and limits. J. Biotechnol. 1999; 75: 291-295. Ref.: https://tinyurl.com/y4cltzsy
Huggett J, Dheda K, Bustin S, Zumla A. Real-time RT-PCR normalization: strategies and considerations. Genes Immun. 2006; 6: 279-84. Ref.: https://tinyurl.com/y4wlytm4
Tian C, Jiang Q, Wang F, Wang GL, Xu ZS, et al. Selection of suitable reference genes for qPCR normalization under abiotic stresses and hormone stimuli in carrot leaves. Plos One. 2015; 10: e0117569. Ref.: https://tinyurl.com/y6e4nqcs
Santi S, Schmidt W. Laser microdissection-assisted analysis of the functional fate of iron deficiency-induced root hairs in cucumber. J Exp Bot. 2008; 59:697-704. Ref.: https://tinyurl.com/y3u8yhj3
Xia XJ, Zhang Y, Wu JX, Wang JT, Zhou YH, et al. Brassinosteroids promote metabolism of pesticides in cucumber. Agric Food Chem. 2009; 57: 8406-8413. Ref.: https://tinyurl.com/y3mxyjnd
Robinson TL, Sutherland IA, Sutherland J. Validation of candidate bovine reference genes for use with real-time PCR. Vet Immunol Immunopathol. 2007; 115: 160-165. Ref.: https://tinyurl.com/y6hpdq7j
Migocka M, Papierniak A. Identification of suitable reference genes for studying gene expression in cucumber plants subjected to abiotic stress and growth regulators. Molecular Breeding. 2011; 28: 343-357. Ref.: https://tinyurl.com/y5af57ng
Chandna R, Augustine R, Bisht NC. Evaluation of candidate reference genes for gene expression normalization in Brassica juncea using real time quantitative RT-PCR. PLoS One. 2012; 7: e36918. Ref.: https://tinyurl.com/y3n85cka
Hu R, Fan C, Li H, Zhang Q, Fu YF. Evaluation of putative reference genes for gene expression normalization in soybean by quantitative real-time RT-PCR. BMC Mol Biol. 2009; 10:93. Ref.: https://tinyurl.com/y2dzlz8y
Warzybok A, Migocka M. Reliable reference genes for normalization of gene expression in cucumber grown under different nitrogen nutrition. Plos One. 2013; 8: e72887 Ref.: https://tinyurl.com/y67f7g8m
Wan H, Zhao Z, Qian C, Sui Y, Chen J. Selection of appropriate reference genes for gene expression studies by quantitative real-time polymerase chain reaction in cucumber. Anal Biochem. 2009; 399: 257-261. Ref.: https://tinyurl.com/y3ayy86k
Bustin SA. Quantification of mRNA using real-time reverse transcription PCR (RT-PCR): trends and problems. J Mol Endocrinol. 2002; 29: 23-39. Ref.: https://tinyurl.com/y3nrrjmo
Silver N, Best S, Jiang J, Thein SL. Selection of housekeeping genes for gene expression studies in human reticulocytes using real-time PCR. BMC Mol Biol. 2006; 7: 33. Ref.: https://tinyurl.com/y6zlcchr
Pfaffl MW, Tichopad A, Prgomet C, Neuvians TP. Determination of stable housekeeping genes, differentially regulated target genes and sample integrity: BestKeeper-excel-based tool using pair-wise correlations. Biotechnol Lett. 2004; 26: 509-515. Ref..: https://tinyurl.com/y4y3frkb
Andersen CL, Jensen JL, Orntoft TF. Normalization of real-time quantitative reverse transcription-PCR data: a model-based variance estimation approach to identify genes suited for normalization, applied to bladder and colon cancer data sets. Cancer Res. 2004; 64: 5245-5250. Ref.: https://tinyurl.com/yya9s2cj
Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, et al. Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol. 2012; 18: 3 (RESEARCH0034). Ref.: https://tinyurl.com/yy2kw3pu
Song Y, Du D, Chu M, Li X, Xu H, et al. Cloning and characterization of a cucumber phospholipase D gene in response to excess nitrate and other abiotic stresses. Scientia Horticulturae. 2012; 135:128-136. Ref.: https://tinyurl.com/y3f92yns
Wang R, Chen M, Liao F, Jiang F, Ma B, et al. Cloning of the gene encoding cucumber lumazine synthase and an analysis of its promoter activity in cucumber. Canadian Journal of Plant Science. 2010; 90: 809-818. Ref.: https://tinyurl.com/yy97ybos
Wang M, Bi H, Liu P, Ai X. Molecular cloning and expression analysis of the gene encoding sedoheptulose-1,7-bisphosphatase from Cucumis sativus. Scientia Horticulturae. 2011; 129: 414-420. Ref.: https://tinyurl.com/y5937kcp
Qi J, Liu X, Shen D, Miao H, Xie B, et al. A genomic variation map provides insights into the genetic basis of cucumber domestication and diversity. Nat Genet. 2013; 45: 1510-1515. Ref.: https://tinyurl.com/y3hn9wys
Chandrasekaran J, Brumin M, Wolf D, Leibman D, Klap C, et al. Development of broad virus resistance in non-transgenic cucumber using crispr/cas9 technology. Mol Plant Pathol. 2016; 17: 1140-1153. Ref.: https://tinyurl.com/y6rngnpp
Huang S, Li R, Zhang Z, Li L, Gu X, Fan W, et al (2009) The genome of the cucumber, Cucumis sativus L. Nat Genet. 2009; 475: 1275-1281. Ref.: https://tinyurl.com/y2xzsvp9
Wu T, Wang R, Xu X, He X, Sun B, et al. Cucumis sativus L-type lectin receptor kinase (CsLecRK) gene family response to Phytophthora melonis, Phytophthora capsici and water immersion in disease resistant and susceptible cucumber cultivars. Gene. 2014; 549: 214-222. Ref.: https://tinyurl.com/yxsmlxse
Xu X, Wang R, Chao J, Lin Y, Jin Q, et al. The expression patterns of Cucumis sativus WRKY (CsWRKY) family under the condition of inoculation with Phytophthora melonisin disease resistant and susceptible cucumber cultivars. Canadian Journal of Plant Science. 2015; 95: 1121-1131. Ref.: https://tinyurl.com/yycqkswq