中国烟草学报2025，Vol.31Issue(2)：93-103,11.DOI:10.16472/j.chinatobacco.2024.T0086
烟草苯丙氨酸解氨酶基因NtPAL的克隆及功能分析
Cloning and functional analysis of the phenylalanine ammonia-lyase gene NtPAL in tobacco
摘要
Abstract
Tobacco phenylalanine ammonia-lyase(PAL)participates in the phenylalanine metabolism pathway,which is related to tobacco aroma quality and plays a role in defense functions,including disease and stress resistance.The aim is to identify the gene function of phenylalanine ammonia-lyase in the pathway of resistance to tobacco mosaic virus(TMV).To identify the gene function of phenylalanine ammoniase involved in tobacco anti-mosaic pathway,the NtPAL gene was identified and cloned from cultivated tobacco(cv.:Hong Hua Da Jin Yuan)by homologous cloning.The protein structures and evolutionary relationships of NtPAL in tobacco were analyzed using bioinformatics method.CRISPR/cas9 gene editing technology was employed to obtain NtPAL gene-edited plants(PAL-KO),and polyphenol content and PAL activity were measured at the plant's mature stage.Subcellular localization of NtPAL was analyzed using a transient expression system in Nicotiana benthamiana.The protein coding sequence of the tobacco NtPAL gene is 2016bp,encoding a PAL protein consisting of 671 amino acids.NtPAL contains a typical phenylalanine ammonia-lyase domain(PLN02457),identifying it as a novel NtPAL gene in tobacco.Subcellular localization experiments showed that NtPAL is localized in the nucleus.Compared with CK,,the PAL-KO edited plants showed reduced resistance to TMV.In mature leaves,the contents of five polyphenols—neochlorogenic acid,scopoletin,cryptochlorogenic acid,chlorogenic acid,and rutin—were significantly increased,while PAL activity was decreased.Tobacco NtPAL responds to TMV resistance by regulating polyphenols content and phenylalanine ammoniase activity.The results of this study provide a theoretical basis for further elucidating the potential of NtPAL in enhancing disease resistance and genetic improvement of tobacco.关键词
烟草/NtPAL/亚细胞定位/基因功能/抗病性Key words
Nicotiana tabacum/NtPAL/subcellular localization/gene function/disease resistance引用本文复制引用
许力,许永,向海英,黄海涛,米其利,陈钰栋,蒋佳芮,李雪梅,高茜..烟草苯丙氨酸解氨酶基因NtPAL的克隆及功能分析[J].中国烟草学报,2025,31(2):93-103,11.基金项目
中国烟草总公司科技项目"多酚类物质代谢相关新品系配方适用性评价及应用"(110202201015(JY-15)) （110202201015(JY-15）
云南大观实验室课题、云南中烟工业有限责任公司科技项目"低芸香苷新品系的品质保障技术优化及烟叶配伍性研究"(YNDG202301YZ01、2023YJ02) （YNDG202301YZ01、2023YJ02）
