摘要
过氧化物酶(peroxidase, PRX)是过氧化物酶体的标志酶,能够保护细胞免受氧化损伤和解除H2O_(2)的毒害作用,还可以增强自然杀伤细胞的活性,调节细胞的增殖、分化和凋亡等.其中Ⅲ类过氧化物酶(CⅢPRXs)是植物中特有的过氧化物酶家族,通过清除活性氧(ROS)在植物免疫中发挥重要作用,然而CⅢPRXs在水稻株型建立中的功能尚不清楚.通过基因编辑技术CRISPR/Cas9获得CⅢPRXs基因(LOC_Os12g09460)两种不同形式的敲除突变体iph1-1,iph1-2.iph1突变体的株高显著高于野生型,除倒1节节长(即第5节)外,其他节节长均显著或极显著长于野生型.农艺性状考察表明,穗长及结实率等主要性状差异无统计学意义;通过RT-qPCR技术进行的表达模式分析表明,IPH1在根、茎、叶、鞘、穗中均表达,并在茎和鞘中表达相对较高;亚细胞定位分析表明,IPH1蛋白主要定位于过氧化物酶体中.进一步通过生理分析,发现突变体中过氧化物酶活性显著降低,同时H2O_(2)质量分数显著增加.这些结果初步证实了IPH1能够通过过氧化氢途径调控水稻株高的形成,可为丰富株高调控网络提供有利的基因资源,进一步为株型相关生物育种奠定基础.
Peroxidase(PRX)is a marker enzyme of peroxisome,which can protect cells from oxidative damage and relieve the toxic effect of H2O_(2).It can also enhance the activity of natural killer cells and regulate cell proliferation,differentiation and apoptosis.The classⅢperoxidase(CⅢPRXs)is a unique peroxidase family in plants,which plays an important role in plant immunity by scavenging reactive oxygen species(ROS).However,the function of CⅢPRXs in the establishment of plant architecture remains unclear in rice.In this study,two different forms of knockout homozygous mutants iph1-1 and iph1-2 of CⅢPRXs gene(LOC_Os12g09460)were obtained by CRISPR/Cas9.The plant height of the iph1 mutant was significantly higher than that of the wild type.Except for the length of the last internode,the lengths of other internodes were significantly or extremely significantly longer than that of the wild type.The investigation of agronomic traits showed that there were no significant differences in main traits such as panicle length and seed setting rate.The expression pattern analyzed by RT-qPCR showed that IPH1 was expressed in roots,stems,leaves,sheaths and panicles,and was relatively highly expressed in stems and sheaths.Subcellular localization analysis showed that IPH1 was mainly located in peroxisomes.Through further physiological analysis,it was found that the peroxidase activity in iph1 mutant was significantly reduced,while the H2O_(2) content was increased.These results preliminarily confirmed that IPH1 regulates the formation of plant height through the hydrogen peroxide pathway in rice,which can provide favorable gene resources for enriching the plant height regulation network and further lay a good foundation for plant architecture related biological breeding.
作者
冯萍
刘杨
杨杰
马宏蕾
秦诚
王楠
沈文强
FENG Ping;LIU Yang;YANG Jie;MA Honglei;QIN Cheng;WANG Nan;SHEN Wenqiang(Rice Research Institute,Southwest University/Academy of Agricultural Sciences/Chongqing Key Laboratory of Transgenic Plant and Safety Control,Chongqing 400715,China)
出处
《西南大学学报（自然科学版）》
CSCD
北大核心
2024年第2期24-33,共10页
Journal of Southwest University(Natural Science Edition)
基金
重庆市技术创新与应用发展专项(CSTB2022TIAD-KPX0014)
重庆市自然科学基金项目(cstc2021jcyj-bshX0076)
西南大学实验技术研究项目(SYJ2022025).
