青花菜BolPEN2基因克隆与抗软腐病功能研究  

Cloning and Functional Analysis of BolPEN2 Gene against Soft Rot in Broccoli

作　　者：马超超 王瑾 王尧 陶婧婷 王春国[2] 李慧 MA Chaochao;WANG Jin;WANG Yao;TAO Jinting;WANG Chunguo;LI Hui(College of Horticulture and Landscape,Tianjin Agriculture University,Tianjin 300392;College of Life Sciences,Nankai University,Tianjin 300071)

机构地区：[1]天津农学院园艺园林学院,天津300392 [2]南开大学生命科学学院,天津300071

出　　处：《核农学报》2025年第4期686-695,共10页Journal of Nuclear Agricultural Sciences

基　　金：天津市科技支撑重点项目(23YFZCSN00190)。

摘　　要：硫代葡萄糖苷(硫苷)是青花菜等十字花科植物产生的一类重要次生代谢产物,这其中吲哚族硫苷及其水解产物被证实在植物抵御病原菌侵染等生物胁迫中发挥重要作用。PEN2是调控吲哚族硫苷水解的关键基因,为了探究该类基因在青花菜硫苷水解及响应生物胁迫中的功能,本研究对青花菜中PEN2基因(BolPEN2)进行了克隆,并对其转录表达特征及其在响应软腐病菌侵染胁迫中的功能进行了研究。结果表明,BolPEN2编码区全长1668 bp,编码555个氨基酸。BolPEN2在种子中呈现高表达,并且在青花菜受到软腐病菌侵染胁迫后其表达水平呈现上升趋势,暗示BolPEN2在青花菜响应生物胁迫中发挥重要作用。为进一步探究BolPEN2的功能,成功创制了BolPEN2过表达及RNAi沉默青花菜转化材料。生理及表型分析结果表明,BolPEN2过表达显著提高了青花菜转化株对软腐病的抗性,BolPEN2沉默青花菜材料则表现出更强的感病特征。与之相对应,BolPEN2过表达青花菜植株叶片总硫苷含量显著低于对照青花菜,而BolPEN2沉默青花菜植株中总硫苷含量显著高于对照。研究结果证实BolPEN2可能通过调控吲哚族硫苷水解过程而在青花菜抗软腐病中发挥重要作用,该结果为进一步揭示BolPEN2的功能及通过基因工程手段创制抗病青花菜材料奠定了基础。Glucosinolates(GSLs)are a class of important secondary metabolites produced in cruciferous plants such as broccoli.Among them,indole GSLs and their hydrolysates have been confirmed to play important roles in plant resistance to biotic stresses.PEN2 is a key gene regulating the hydrolysis of indole GSLs.However,the roles of PEN2 in regulating the hydrolysis of GSLs and responding to the biotic stresses in broccoli remains unrevealed.In this study,the PEN2 gene(BolPEN2)in broccoli was cloned,and the transcriptional expression characteristics and functions of BolPEN2 in response to soft rot infection stress were studied.The results showed that the coding region of BolPEN2 was 1668 bp in length,encoding 555 amino acids.BolPEN2 was highly expressed in seeds,and its expression level showed an upward trend under soft rot infection stress in broccoli.In order to further explore the function of BolPEN2,transgenic broccoli lines were developed for both BolPEN2 overexpression and RNAi silencing.Physiological and phenotypic analysis showed that overexpression of BolPEN2 significantly improved the resistance of broccoli to soft rot infection,while BolPEN2 silencing lines were more susceptible to soft rot infection than that of the control broccoli.Consistent with these results,the total content of GSLs in leaves of BolPEN2-overexpressingbroccoli lines was significantly lower than that of the control broccoli,while the GSLs content in BolPEN2 silencing plants was significantly higher than that of the control.The results indicated that BolPEN2 may play an important role in increasing tolerance of broccoli to soft rot infection by regulating the hydrolysis process of GSLs,especially indole GSLs.These findings provide new insights into the roles of BolPEN2,and suggest potential applications of BolPEN2 in breeding crops with high tolerance to soft rot infection by genetic engineering.

关 键 词：青花菜 硫代葡萄糖苷 BolPEN2 软腐病 

分 类 号：S635.3[农业科学—蔬菜学]