Yao Qian, Duan Ruiwei, Feng Yang, Duan Dong
Key Laboratory of Resource Biology and Biotechnology in Western China, Ministry of Education, College of Life Sciences, Northwest University, Xi'an, 710069, China.
Planta. 2025 May 14;261(6):139. doi: 10.1007/s00425-025-04713-1.
Alternative splicing of transcriptomes after Al and flg22 treatment for 12 h in response to plant defense of Chinese wild Vitis quinquangularis: genes related to stress resistance and splicing factors were identified in response to Al and flg22 treatment. Alternative splicing (AS) is one of the major post-transcriptional regulation processes that potentially regulates the response to biotic and abiotic stresses in plants. So far, the insight into potential roles of AS in grapevine response to aluminium (Al) and flagellin 22 (flg22) stresses remains poorly understood. We performed transcriptome sequencing of grape leaves before and after Al treatment and flg22 treatment, respectively, to identify AS genes. In this study, a total of 11,805 AS events were identified in Al treatment, of which the skipped exon (SE; 88.72%) type was the most frequent. 9156 AS events were identified under flg22 treatment, of which the SE (88.52%) type was the most frequent. Compared with Al-treated and flg22-treated 0 h, there were 42 and 147 differential alternative splicing (DAS) genes differentially expressed (DASEGs) in Al-treated and flg22-treated 12 h, respectively. Functional analysis showed that DASEGs after Al treatment were mainly enriched in glutathione metabolism pathway; DASEGs after flg22 treatment were enriched in MAPK signaling and plant hormone signal transduction. We further verified seven resistance-related DASEGs with up-regulated expression in Al-treated 12 h, including beta-glucosidase, calcineurin B-like protein, synaptotagmin-3, cysteine synthase and glutathione reductase. Several genes function as leucine-rich repeats receptor-like serine/threonine protein kinase, BRI1 associated receptor kinase 1 and receptor-like protein kinase were also verified by RT-qPCR. We also verified four serine/arginine (SR)-rich proteins SCL30A, SCL28, RS2Z32 and SR45A, which were up-regulated in both Al and flg22 stresses. In conclusion, this study provides an in-depth analysis of the correlation between alternative splicing and grapevine stress tolerance, which helps to identify potential candidate genes for useful traits, provides a theoretical basis for grapevine breeding in plant stress tolerance, and offers new perspectives for understanding grapevine environmental adaptation strategies.
中国野生毛葡萄响应植物防御时，铝（Al）和鞭毛蛋白22（flg22）处理12小时后转录组的可变剪接：鉴定了与抗逆性相关的基因和响应Al与flg22处理的剪接因子。可变剪接（AS）是主要的转录后调控过程之一，可能调控植物对生物和非生物胁迫的响应。到目前为止，对AS在葡萄对铝（Al）和鞭毛蛋白22（flg22）胁迫响应中的潜在作用了解甚少。我们分别对Al处理和flg22处理前后的葡萄叶片进行转录组测序，以鉴定AS基因。在本研究中，Al处理中共鉴定出11,805个AS事件，其中外显子跳跃（SE；88.72%）类型最为常见。flg22处理下鉴定出9156个AS事件，其中SE（88.52%）类型最为常见。与Al处理和flg22处理0小时相比，Al处理12小时和flg22处理12小时分别有42个和147个差异可变剪接（DAS）基因差异表达（DASEGs）。功能分析表明，Al处理后的DASEGs主要富集于谷胱甘肽代谢途径；flg22处理后的DASEGs富集于MAPK信号传导和植物激素信号转导。我们进一步验证了Al处理12小时后上调表达的7个与抗性相关DASEGs，包括β-葡萄糖苷酶、类钙调神经磷酸酶B蛋白、突触结合蛋白-3、半胱氨酸合酶和谷胱甘肽还原酶。几个作为富含亮氨酸重复序列的受体样丝氨酸/苏氨酸蛋白激酶、BRI1相关受体激酶1和受体样蛋白激酶的基因也通过RT-qPCR得到验证。我们还验证了4个富含丝氨酸/精氨酸（SR）的蛋白SCL30A、SCL28、RS2Z32和SR45A，它们在Al和flg22胁迫下均上调表达。总之，本研究深入分析了可变剪接与葡萄抗逆性之间的相关性，有助于鉴定有用性状的潜在候选基因，为葡萄抗逆性育种提供理论依据，并为理解葡萄环境适应策略提供新视角。
