不同组织与逆境胁迫下茄子qRT-PCR内参基因筛选

Screening of Eggplant qRT-PCR Internal Reference Genes under Different Tissues and Adversity Stresses

为了筛选茄子中不同组织与不同逆境胁迫下合适的内参基因,本研究选取UBC、EF-1a、18S、Actin、GAPDH、β-TUB、L33和EF28个候选内参基因,在涝害胁迫、干旱胁迫、冷胁迫、热胁迫、大丽轮枝菌侵染、盐胁迫、水杨酸处理和不同组织部位8种试验条件下,利用geNorm、NormFinder、ΔC_(T)、Bestkeeper和RefFinder对其稳定性进行分析。结果显示,在全部样品、冷胁迫、热胁迫、大丽轮枝菌侵染和盐胁迫下,Actin均为最稳定内参基因,β-TUB是涝害胁迫下表达最稳定的基因,干旱胁迫下表达最稳定的是EF2,水杨酸处理下表达最稳定的是EF-1a,18S在不同组织部位的处理下稳定性最好。UBC、L33和GAPDH在各个处理下均不稳定表达。本研究为茄子不同组织及逆境胁迫下基因特异性表达研究提供了合适的内参基因。

To screen for suitable internal reference genes in eggplant under different tissues and adversity stresses,UBC,EF-1a,18S,Actin,GAPDH,β-TUB,L33 and EF2,eight candidate internal reference genes were selected in this study under eight experimental conditions,including flood stress,drought stress,cold stress,heat stress,Verticillium dahliae infestation,salt stress,salicylic acid treatment and different tissue sites.The stability of candidate internal reference genes was analyzed using geNorm,NormFinder,ΔC_(T),Bestkeeper and RefFinder under eight test conditions.The results showed that Actin was the most stable internal gene among all samples,cold stress,heat stress,Verticillium dahliae infestation and salt stress,β-TUB was the most stable gene expressed under flooding stress,EF2 was the most stable under drought stress,EF-1a was the most stable gene expressed under salicylic acid treatment,and 18S was the most stable among different tissue sites.UBC,L33 and GAPDH were not stably expressed under all treatments.This study provides stable internal reference genes for the study of gene specific expression in different tissues of eggplants and under adversity stress.