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为揭示番茄叶片中miRNA对非生物胁迫的响应机制,以番茄ABA缺失突变体sit和野生型番茄RR为试验材料。采用实时荧光定量(qRT-PCR)技术检测叶片中6个miRNA(miR160、miR162、miR166、miR1919、miR397和miR6026)及其预测的靶基因在夜间低温和喷施外源ABA逆境胁迫下的表达情况。结果表明,在15℃夜温(对照)下,sit的ABA含量是60 ng·g-1,RR是120 ng·g~(-1),sit叶片的保卫细胞和气孔的开度都明显大于野生型RR,ABA缺失突变体sit中miRNA的表达量与野生型RR相比都显著下调,其靶基因除miR1919外,表达量都上调;夜间低温(6℃)处理12 h后,所有miRNA在sit中比RR中的表达量都显著增加;喷施80 mg·L~(-1)的ABA后,sit中miR162、miR166、miR1919及miR397比0 mg·L~(-1) ABA处理的表达量显著上调。推测miR160、miR162和miR6026通过依赖ABA信号途径参与夜间低温胁迫。
To reveal the mechanism of miRNA response to abiotic stresses in tomato leaves, tomato ABA deletion mutant sit and wild-type tomato RR were used as experimental materials. The expression of 6 miRNAs (miR160, miR162, miR166, miR1919, miR397 and miR6026) in leaves and their predicted target genes under low nocturnal and exogenous ABA stress were detected by qRT-PCR . The results showed that the ABA content of sit was 60 ng · g-1 and the RR was 120 ng · g -1 at 15 ℃ overnight, the guard cells and stomata in sit leaves were significantly larger than that in sit leaves The expression of miRNA in wild-type RR and aba-deficient mutant sit was significantly down-regulated compared with wild-type RR, and the expression of all miRNAs was up-regulated except for miR1919. After 12 hours of low temperature (6 ℃) the expression of miR162, miR166, miR1919 and miR397 was significantly higher than that of RR at 0 mg · L -1 sit after spraying ABA of 80 mg · L -1 Significantly increased. It is speculated that miR160, miR162 and miR6026 participate in nocturnal hypothermia through the ABA-dependent signaling pathway.