摘要
【目的】揭示根蘖繁殖与嫁接繁殖对灵武长枣果实可溶性糖代谢变化的差异,为提高果实品质提供科学依据。【方法】以根蘖与嫁接灵武长枣在白熟期(NB vs JB)、着色期(NZ vs JZ)和成熟期(NC vs JC)的果实为材料,测定果实葡萄糖、果糖、蔗糖及可溶性糖含量;基于转录组数据,从糖酵解/糖异生(ko00010)、淀粉与蔗糖代谢(ko00500)、果糖与甘露糖代谢(ko00051)以及氨基糖和核苷酸糖代谢(ko000520)4个关键糖代谢通路中,筛选与可溶性糖合成代谢相关的差异基因;将表型数据与转录组数据结合,进行WGCNA分析;利用qRT-PCR验证以确保结果的可靠性。【结果】灵武长枣在白熟期和着色期的糖分积累形式主要为果糖,而在成熟期则以蔗糖为主要积累形式。不同繁殖方式下灵武长枣果实糖类积累呈现显著的发育阶段特异性。着色期根蘖繁殖的果糖含量比嫁接繁殖高20.25%;成熟期根蘖繁殖的蔗糖含量比嫁接繁殖高49.32%。从上述4条通路中筛选获得了24个显著差异表达基因(DEGs)。通过GO功能富集分析发现,这些差异基因主要富集在碳水化合物代谢过程相关功能条目中。此外,加权基因共表达网络分析结果表明,黑色、棕色和绿色模块与可溶性糖代谢显著关联,其中ncbi_112492650、HERC2(ncbi_107420452)、PGMP(ncbi_107414395)基因的连通性最强。qRT-PCR分析结果表明,所测定的差异表达基因在不同时期的表达模式存在差异,与转录组数据中的表达趋势一致。【结论】基因ncbi_112492650、HERC2(ncbi_107420452)、PGMP(ncbi_107414395)在调控灵武长枣可溶性糖代谢方面发挥关键作用,可作为后续关键验证基因。本研究为优化灵武长枣的繁殖方式及提高果实品质提供了重要的理论依据。
【Objective】Ziziphus jujuba Mill.‘Lingwuchangzao’belongs to Rhamnaceae Jujube.Its fruits are rich in protein,sugar and other nutrients,and have high economic value and market development potential.It uasually propagated by root tillers and grafting.The study aimed to reveal the differences of soluble sugar metabolism in the fruits of Lingwuchangzao propagated by different propagaion methods through studying on the changes of soluble sugar content in different developmental stages of the fruits of Lingwuchangzao propagated by root tillers and grafting in order to provide a scientific basis for improving fruit quality.【Methods】The investigation was carried ou at the white ripening stage(NB vs JB),coloring stage(NZ vs JZ),and mature stage(NC vs JC)on the fruits of Lingwuchangzao propagated by root suckers and grafting as experimental materials,.The contents of glucose,fructose,sucrose and soluble sugar in the fruits of the three periods were determined respectively.Combined with transcriptome data analysis,significant differential genes were screened from four key sugar metabolism pathways,glycolysis/gluconeogenesis(ko00010),starch and sucrose metabolism(ko00500),fructose and mannose metabolism(ko00051),and amino sugar and nucleotide sugar metabolism(ko000520)with FDR<0.05 and|log2FC|≥1 as the standard.By integrating phenotypic data with transcriptome data for WGCNA analysis,key differential genes regulating soluble sugar synthesis and metabolism at different developmental stages of Lingwuchangzao fruits were identified,followed by qRTPCR verification to ensure the reliability of the results.【Results】The soluble sugars in Lingwuchangzao included sucrose,fructose,glucose and others.The fructose was the main sugar accumulated at white ripe stage and coloring stage,while sucrose was the main sugar at mature stage.However,sugar accumulation exhibited significant developmental stage specificity in the fruits of Lingwuchangzao propagated with different methods.At the white ripe stage,the soluble sugar content of the fruits from Lingwuchangzao propagated by grafting was 17.55%higher than that from root tillers.At the coloring stage and maturity stage,the soluble sugar content of fruits from the Lingwuchangzao propagated by root tillers was 24.38%and 27.91%higher than that from grafted trees,respectively.At the coloring and mature stages,the sucrose content in the fruits fruits from Lingwuchangzao propagated by propagated by root tiller was 50.63%and 49.32%higher than that from grafted trees,respectively.At the coloring stage,the fructose content of in the fruits fruits from root tiller propagated trees was 20.25%higher than that from grafted trees.At the mature stage,the fructose content in the fruits from grafted trees was 13.32%higher than that from root tiller propagated trees.As the fruit grew and developed,the glucose content in the fruits from grafting propagated trees was 36.08%,20.67%,and 26.85%higher than from the trees propagated by root tiller,respectively.24 significantly differentially expressed genes(DEGs)were screened.Among them,the genes EP3(ncbi_107406053),BAM1(ncbi_107422617),BGLU11(ncbi_107426268),PGMP(ncbi_107414395)showed higher expression level,and the difference of the gene expressions between the coloring period and the maturity period was significant,indicating the potential for further fluorescence quantitative analysis.Through GO functional enrichment analysis,it was found that there were significant differences in carbohydrate metabolism,hydrolytic enzyme activity and secondary metabolite synthesis between the trees propagated by two different methods.These differences might be closely related to the adaptability of trees and the fruit quality.The analysis of soluble sugar metabolism pathways and differential genes revealed that the gene PGMP(ncbi_107414395)appeared multiple times in the metabolic pathways,indicating that this gene might play an important role in regulating soluble sugars in Lingwuchangzao and could serve as a key candidate gene for further validation studies.Additionally,weighted gene co-expression network analysis(WGCNA)showed that the black,brown,and green modules were significantly associated with soluble sugar metabolism.Genes in the black module exhibited a strong positive correlation with fructose,with a correlation coefficient of 0.87;genes in the brown and green modules showed strong positive correlations with soluble sugar and sucrose,with correlation coefficients of 0.71,0.86,0.74,and 0.83,respectively.Finally,the genes ncbi_112492650 and HERC2(ncbi_107420452),which exhibited the highest connectivity and high expression level in the black module,were selected as key candidate genes for subsequent validation.Through the fluorescence quantitative analysis(qRT-PCR),it was found that the trend of gene expression levels in the fruits from the trees grafted by root-tiller was basically consistent with the transcriptome sequencing results at the white ripe stage,coloring stage and mature stage,indicating that the transcriptome data had high reliability.【Conclusion】This study revealed the difference of soluble sugar metabolism in the fruits of Lingwuchangzao propagated by two differnt methods,providing an important theoretical basis for optimizing the propagation mode of Lingwuchangzao.The genes ncbi_112492650,HERC2(ncbi_107420452)and PGMP(ncbi_107414395)would play a key role in regulating soluble sugar metabolism of Lingwuchangzao,and could be used as subsequent key verification genes.
作者
李思思
杨荣
刘佳嘉
何开明
刘思佳
LI Sisi;YANG Rong;LIU Jiajia;HE Kaiming;LIU Sijia(College of Forestry and Grassland,Ningxia University,Yinchuan 750021,Ningxia,China;National Key Laboratory of Efficient Production of Forest Tree Resources,Yinchuan 750021,Ningxia,China;Lingwu City Natural Resources Bureau,Yinchuan 750021,Ningxia,China)
出处
《果树学报》
北大核心
2026年第2期309-322,共14页
Journal of Fruit Science
基金
国家自然科学基金项目(32060338)
北京林业大学中央高校基本科研业务费专项资金项目(BFKUF202401)。
关键词
灵武长枣
根蘖繁殖
嫁接繁殖
可溶性糖
差异基因
Ziziphus jujuba Mill.‘Lingwuchangzao’
Root tiller propagation
Grafting propagation
Soluble sugars
Differential genes
