Beijing Key Laboratory for Agricultural Application and New Technology, National Demonstration Center for Experimental Plant Production Education, College of Plant Science and Technology, Beijing University of Agriculture, Beijing, China.
University of Arizona, Tucson, Arizona, USA.
Plant Signal Behav. 2021 Jul 3;16(7):1913845. doi: 10.1080/15592324.2021.1913845. Epub 2021 May 6.
To determine the effect of the serine/threonine protein kinase (STPK) gene on leaf lettuce bolting, we utilized virus-induced gene silencing (VIGS) using the TRV vector to silence the target gene. The 'GB30' leaf lettuce cultivar was the test material, and the methods included gene cloning, bioinformatics analysis, quantitative real-time PCR (qRT-PCR) and VIGS. LsSTPK, was cloned from the 'GB30' leaf lettuce cultivar via reverse transcription-polymerase chain reaction (RT-PCR). qRT-PCR analysis showed that the expression of LsSTPK in the stem of leaf lettuce was significantly greater than that in the roots and leaves, and after high-temperature treatment, the gene expression in the stems in the experimental group was markedly lower than that in the control groups. Following LsSTPK silencing via the VIGS method, the stem length in the treatment group was significantly greater than that in the blank and negative control groups, and the contents of auxin (IAA), GA3 and abscisic acid (ABA) in the treatment group were greater than those in the other two groups. Flower bud differentiation occurred in the treatment group but not in the control group. The above findings suggested that LsSTPK inhibits the bolting of leaf lettuce under high-temperature conditions.
为了确定丝氨酸/苏氨酸蛋白激酶(STPK)基因对生菜抽薹的影响,我们利用 TRV 载体的病毒诱导基因沉默(VIGS)来沉默靶基因。试验材料为‘GB30’生菜品种,方法包括基因克隆、生物信息学分析、定量实时 PCR(qRT-PCR)和 VIGS。通过反转录-聚合酶链反应(RT-PCR)从‘GB30’生菜品种中克隆了 LsSTPK。qRT-PCR 分析表明,LsSTPK 在生菜茎中的表达明显高于根和叶,高温处理后,实验组茎中的基因表达明显低于对照组。通过 VIGS 法沉默 LsSTPK 后,处理组的茎长明显大于空白组和阴性对照组,处理组的生长素(IAA)、GA3 和脱落酸(ABA)含量均大于其他两组。处理组出现花芽分化,但对照组没有。上述结果表明,LsSTPK 抑制生菜在高温条件下的抽薹。