本文選題：芝麻莖點(diǎn)枯病菌 + qRT-PCR��； 參考：《中國油料作物學(xué)報(bào)》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(shí)時(shí)熒光定量PCR分析的內(nèi)參基因,11個(gè)基因作為候選內(nèi)參基因。經(jīng)過PCR擴(kuò)增效率篩選,β-肌動(dòng)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(nèi)部轉(zhuǎn)錄間隔區(qū)(ITS)8個(gè)基因符合要求,可用于穩(wěn)定度篩選。利用實(shí)時(shí)熒光定量PCR技術(shù),檢測了這8個(gè)候選基因在菜豆殼球孢侵染芝麻8h、16h、24h和32h的表達(dá)情況。經(jīng)Ge Norm軟件分析,ACTB、GAPDH和RPS5-b等3個(gè)基因表達(dá)較穩(wěn)定。經(jīng)過最適內(nèi)參基因數(shù)目分析,在菜豆殼球孢侵染芝麻過程中基因定量表達(dá)分析時(shí),選擇ACTB、GAPDH和RPS5-b的多基因組合作為內(nèi)參,能夠更準(zhǔn)確地校正定量結(jié)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(nóng)業(yè)科學(xué)院植物保護(hù)研究所農(nóng)業(yè)部華北南部農(nóng)作物有害生物綜合治理重點(diǎn)實(shí)驗(yàn)室河南省農(nóng)作物病蟲害防治重點(diǎn)實(shí)驗(yàn)室;
【基金】：國家自然科學(xué)基金(31301631) 農(nóng)業(yè)部現(xiàn)代農(nóng)業(yè)產(chǎn)業(yè)技術(shù)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(guān)系[J];植物保護(hù)學(xué)報(bào);1986年03期

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