本文選題：芝麻莖點枯病菌 + qRT-PCR　； 參考：《中國油料作物學報》2017年03期

【摘要】：為選擇適合菜豆殼球孢(Macrophomina phaseolina)侵染芝麻過程中實時熒光定量PCR分析的內參基因,11個基因作為候選內參基因。經過PCR擴增效率篩選,β-肌動蛋白(ACTB)、泛素連接酶(UBC)、α-微管蛋白(TUBA)、γ-微管蛋白(TUBC)、3-磷酸甘油醛脫氫酶基因(GAPDH)、核糖體蛋白S5(RPS5-a,RPS5-b)和內部轉錄間隔區(qū)(ITS)8個基因符合要求,可用于穩(wěn)定度篩選。利用實時熒光定量PCR技術,檢測了這8個候選基因在菜豆殼球孢侵染芝麻8h、16h、24h和32h的表達情況。經Ge Norm軟件分析,ACTB、GAPDH和RPS5-b等3個基因表達較穩(wěn)定。經過最適內參基因數目分析,在菜豆殼球孢侵染芝麻過程中基因定量表達分析時,選擇ACTB、GAPDH和RPS5-b的多基因組合作為內參,能夠更準確地校正定量結果。
[Abstract]:In order to select the internal reference gene suitable for the real-time fluorescence quantitative PCR analysis of sesame seeds infected by Macrophomina phaseolina, 11 genes were used as candidate internal reference genes. After PCR amplification, eight genes, 尾 -actin ACTBN, ubiquitin ligase (UBCU), 偽 -tubulin (TUBAA), 緯 -tubulin (TUBC) 3-phosphoglyceraldehyde dehydrogenase (GAPDHN), ribosomal protein S5 (RPS5-aRPS5-b) and internal transcriptional spacer (ITS5), met the requirements and could be used for stability screening. The expression of these eight candidate genes was detected by real-time fluorescence quantitative PCR (PCR) in cultured sesame seeds infected with Phaseolus vulgaris for 24 h and 32 h, respectively. The expression of GAPDH and RPS5-b were stable by GE Norm software. According to the analysis of the optimum number of internal reference genes, the multi-genome cooperation of RPS5-b and ACTB GAPDH can be used to correct the quantitative results more accurately in the process of quantitative expression analysis of the genes in the process of infecting sesame seeds.
【作者單位】： 河南省農業(yè)科學院植物保護研究所農業(yè)部華北南部農作物有害生物綜合治理重點實驗室河南省農作物病蟲害防治重點實驗室;
【基金】：國家自然科學基金(31301631) 農業(yè)部現代農業(yè)產業(yè)技術體系(CARS-15-1-05)
【分類號】：S435.653

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1 李慶基;L. S. Bird;P. M. Thaxton;M. L. Howell;;菜豆殼球孢菌與棉花的關系[J];植物保護學報;1986年03期

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本文編號：1853084