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Abstract：
This study systematically investigated the physiological and molecular responses of the wheat mutant \'XC-MU201\' under high-temperature stress through comprehensive transcriptome analysis and physiological measurements. RNA sequencing of 21 samples across seven different treatment groups revealed, through Weighted Gene Co-expression Network Analysis (WGCNA), 13 modules among 9071 genes closely related to high-temperature treatments. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analyses showed significant enrichment of lignin biosynthesis-related modules under high-temperature conditions, especially at the H-10DAT, H-20DAT, and H-30DAT time points. Experimental results demonstrated a significant increase in lignin content in high-temperature-treated samples, confirmed by tissue staining methods, indicating wheat\'s adaptation to heat damage through lignin accumulation. The phenylalanine ammonia-lyase gene (TaPAL33) was significantly upregulated under high-temperature stress, peaking at H-30DAT, suggesting its critical role in cellular defense mechanisms. Overexpression of TaPAL33 in the wheat variety \'Xinchun 11\' enhanced lignin synthesis but inhibited growth. Subcellular localization of GFP-labeled TaPAL33 in tobacco cells showed its distribution mainly in the cytoplasm and cell membrane. Transgenic wheat exhibited higher PAL enzyme activity, enhanced antioxidant defense, and reduced oxidative damage under high-temperature stress, outperforming wild-type wheat. These results highlight TaPAL33\'s key role in improving wheat heat tolerance and provide a genetic foundation for future research and applications.
摘要：
本研究通过综合转录组分析和生理测量，系统地研究了小麦突变体\'XC-MU201\'在高温胁迫下的生理和分子响应。七个不同治疗组的21个样本的RNA测序显示，通过加权基因共表达网络分析(WGCNA)，9071个基因中有13个模块与高温处理密切相关。基因本体论（GO）和京都基因和基因组百科全书（KEGG）途径分析显示，在高温条件下木质素生物合成相关模块的显着富集，尤其是在H-10DAT,H-20DAT,和H-30DAT时间点。实验结果表明，高温处理样品中木质素含量显着增加，通过组织染色方法证实，表明小麦通过木质素积累适应热损伤。苯丙氨酸解氨酶基因(TaPAL33)在高温胁迫下显著上调,在H-30DAT达到峰值，表明其在细胞防御机制中的关键作用。小麦品种“新春11”中TaPAL33的过表达增强了木质素的合成，但抑制了生长。GFP标记的TaPAL33在烟草细胞中的亚细胞定位显示其主要分布在细胞质和细胞膜上。转基因小麦表现出更高的PAL酶活性，增强抗氧化防御，减少高温胁迫下的氧化损伤，优于野生型小麦。这些结果突出了TaPAL33在提高小麦耐热性方面的关键作用,为今后的研究和应用提供了遗传基础。
