摘要
为了探究赤霉素(GA)促进银杏种子萌发的分子机制,本研究在MS培养基和添加有5 mg·L^(-1)GA的MS培养基上培养15 d后,对银杏胚的形态及种子的相关生理生化指标进行比较,采用转录组测序技术鉴定差异表达基因(DEGs)。对DEGs所关联的生理功能及代谢途径进行分析,确定了参与碳水化合物代谢途径的关键基因,并对Gb_33733基因进行了功能验证。研究结果显示,GA显著提高了银杏胚的长度、重量、发芽势和发芽率;经过15 d的萌发,种子中的淀粉和蛋白质含量下降,而还原糖含量及α-淀粉酶活性显著上升。GO分析结果揭示,两种处理条件下特异性调控的179个DEGs主要富集于光合作用、光照强度感知、光质适应及对生物和非生物胁迫的响应等生理过程。KEGG通路分析表明,DEGs主要富集于碳水化合物代谢、氨基糖与核苷酸糖代谢、光合作用、丝裂原活化蛋白激酶(MAPK)信号传导、核质转运及能量代谢等途径。通过蛋白质互作筛选,鉴定了碳水化合物代谢途径中的6个关键基因。此外实时荧光定量PCR(qRT-PCR)结果证实,GA处理显著增强了Gb_33733、Gb_33736、Gb_19037、Gb_01010、Gb_05593和Gb_35011基因的表达水平。而且,Gb_33733基因的异源表达促进了拟南芥种子的萌发。本研究为进一步阐明GA诱导银杏种子萌发的分子基础提供了理论依据。
In order to investigate the molecular mechanism of gibberellin(GA)promoting Ginkgo seed germination,this study compared the morphology of Ginkgo embryos and related physiological and biochemical indicators of seeds after 15 days of cultivation on MS medium or MS medium supplemented with 5 mg L^(-1)GA,Differentially expressed genes(DEGs)were identified using transcriptome sequencing technology.The physiological activities and metabolic pathways associated with the DEGs were analyzed,key genes involved in carbohydrate metabolism pathways were identified,and the functionality of Gb_33733 gene was validated.The results showed that GA significantly increased the length,weight,germination potential,and germination rate of Ginkgo biloba embryos.After 15 days of germination,the starch and protein content in the seeds decreased,while reducing sugar content and alpha amylase activity increased significantly.GO analysis revealed that 179 DEGs under two treatment conditions were mainly enriched in physiological processes such as photosynthesis,light intensity perception,light quality adaptation,and response to biotic and abiotic stress.The KEGG pathway analysis revealed that DEGs were mainly enriched in carbohydrate metabolism,amino sugar and nucleotide sugar metabolism,photosynthesis,mitogen activated protein kinase(MAPK)signaling,nuclear cytoplasmic transport,and energy metabolism pathways.Six important genes involved in carbohydrate metabolism pathways were identified by protein interaction screening.In addition,qRT PCR results confirmed that GA treatment significantly enhanced the expression levels of Gb_33733,Gb_33736,Gb_19037,Gb_01010,Gb_05593,and Gb_35011 genes.Moreover,the heterologous expression of the Gb_33733 gene promotes the germination of Arabidopsis seeds.This study provides a theoretical foundation for further elucidating the molecular mechanism of GA-induced germination of Ginkgo biloba seeds.
作者
张斌
袁志辉
ZHANG Bin;YUAN Zhihui(College of Chemistry and Bioengineering,Hunan University of Science and Engineering,Yongzhou,Hunan 425199)
出处
《核农学报》
CAS
CSCD
北大核心
2024年第11期2074-2082,共9页
Journal of Nuclear Agricultural Sciences
基金
永州市指导性科技计划项目(2020-YZKJ-001)
湖南省自然科学基金项目(2024JJ7190)。
