香榧胚性愈伤组织的悬浮培养动力学及对赤霉素的响应  

Suspension culture dynamics of embryogenic callus from Torreya grandis‘Merrillii’and its response to gibberellin

作　　者：杨正宇 周佳君 胡恒康[1,2] 张启香[1,2] YANG Zhengyu;ZHOU Jiajun;HU Hengkang;ZHANG Qixiang(College of Forestry and Biotechnology,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China;State Key Laboratory of Subtropical Silviculture,Zhejiang A&F University,Hangzhou 311300,Zhejiang,China)

机构地区：[1]浙江农林大学林业与生物技术学院,浙江杭州311300 [2]浙江农林大学省部共建亚热带森林培育国家重点实验室,浙江杭州311300

出　　处：《浙江农林大学学报》2025年第1期103-111,共9页Journal of Zhejiang A&F University

基　　金：浙江省重点研发计划项目(2021C02037)。

摘　　要：【目的】以香榧Torreya grandis ‘Merrillii’胚性愈伤组织为试验材料,对其悬浮细胞的生长以及悬浮培养动力学进行研究,探究香榧胚性愈伤组织对赤霉素的响应。【方法】以实验室培育的香榧胚性愈伤组织为试验材料,通过测定细胞活力、细胞鲜质量进行悬浮培养动力学的研究,优化培养条件,并对施加外源赤霉素A3 (GA3)的愈伤组织进行转录组分析。【结果】(1)在培养周期内,悬浮细胞的鲜质量增长呈“S”型曲线,细胞活力在3 d时达最高值后平稳下降。(2)香榧悬浮细胞培养最佳继代周期为9~12 d,最佳接种量为30 g·L^(-1),培养基最佳初始pH为5.7,最佳摇床转速为110r·min^(-1)。(3)对外源GA3处理后的香榧胚性愈伤组织进行转录组分析,共获得差异表达基因428个,其中上调基因236个,下调基因192个。在差异基因的GO富集分析中,GA3处理后上调表达基因主要参与催化活性、氧化还原过程、氧化还原酶活性等,在KEGG代谢通路中,差异表达基因主要富集在苯丙烷类生物合成、木栓质生物合成等途径中。【结论】香榧胚性愈伤组织培养动力学分析能获得最佳接种时期,优化培养条件能提高其细胞增长率及抗逆性,外源GA3处理下的香榧胚性愈伤组织细胞中,苯丙烷生物合成、木栓质生物合成等代谢通路为差异基因主要参与部分,且部分关键基因表达量变化可能在香榧愈伤组织生长发育、适应环境及应对胁迫中发挥重要作用。[Objective]This study,with embryogenic callus from Torreya grandis‘Merrillii’employed as the experimental material,is aimed to investigate the growth and suspension culture dynamics of its suspension cells,explore the response of T.grandis‘Merrillii’embryogenic callus to gibberellin treatment.[Method]Using the embryogenic callus of T.grandis‘Merrillii’cultivated in laboratory as the experimental material,the suspension culture dynamics were studied by measuring cell viability and fresh cell weight,and the culture conditions were optimized before a transcriptome analysis was performed on the callus treated with exogenous GA3.[Result](1)Within the culture cycle,the fresh weight growth curve of the suspension cells exhibited an“S”shaped pattern,with cell viability reaching its peak at 3 days and then gradually decreasing steadily.(2)The optimal subculture period for T.grandis‘Merrillii’suspension cells was determined to be 9 to 12 days,with an optimal inoculation rate of 30 g·L^(-1),an initial pH of 5.7 for the culture medium,and an optimal shaking speed of 110 r·min^(-1).(3)transcriptome analysis of T.grandis‘Merrillii’embryogenic callus treated with exogenous GA3 revealed 428 differentially expressed genes,including 236 upregulated and 192 downregulated genes whereas GO enrichment analysis of these differentially expressed genes showed that the upregulated genes after GA3 treatment were mainly involved in catalytic activity,oxidation-reduction processes,and oxidoreductase activity,and in the KEGG metabolic pathways,the differentially expressed genes were primarily enriched in phenylpropanoid biosynthesis and suberin biosynthesis.[Conclusion]The kinetic analysis of T.grandis‘Merrillii’embryogenic callus culture can identify the optimal inoculation period and optimizing the culture conditions can enhance cell growth rate and stress resistance.In embryogenic callus of T.grandis‘Merrillii’under exogenous GA3 treatment,metabolic pathways such as phenylpropanoid biosynthesis and suberin

关 键 词：香榧 胚性愈伤组织 悬浮培养 赤霉素 

分 类 号：S722.3[农业科学—林木遗传育种]