1.College of Horticulture and Gardening,Yangtze University;2.Xiangxin Crop Research Institute,Yangtze University;3.College of Agriculture,Yangtze University
Key R&D Program of Hubei Province（2021BBA097；2022BBA0061）；Jingzhou Science and Technology Plan for 2022（2022BB36）；Chongqing Outstanding Scientist Program（2022CQYC0167）
异戊烯基转移酶(isopentenyl-transferases,IPT)是催化细胞分裂素生物合成的关键酶,也是重要的限速酶。在植物中,IPT以ATP/ADP IPTs和tRNA-IPTs的形式存在,其主要介导植物体内两种类型CKs(细胞分裂素)的合成途径：ATP/ADP途径和tRNA途径。生姜是一种重要的“药食同源”植物,但其种植过程中容易受到各种生物/非生物胁迫的影响,不利于生姜的安全生产。本文通过系统的生物信息学分析,从生姜基因组中鉴定到10个ZoIPT,并且将其命名为ZoIPT1-ZoIPT10。基因结构分析表明所有的ZoIPTs基因家族成员均包含内含子。其编码蛋白的氨基酸长度范围在283(ZoIPT8)~491(ZoIPT10) aa之间,分子质量在31.14(ZoIPT8)~54.02(ZoIPT10) kD,等电点在4.97(ZoIPT8)~9.37(ZoIPT6)之间；蛋白质特征分析表明,所有ZoIPTs均具亲水性。亚细胞定位分析表明,8个ZoIPTs定位在叶绿体中,2个ZoIPTs定位在线粒体和细胞质中。转录组数据分析结果显示,ZoIPTs有一定的组织表达特异性,并且能响应病害、低温等逆境胁迫,其中,ZoIPT3和ZoIPT5在生姜不同生长时期、不同部位、低温和病害胁迫下均有较高表达。qRT-PCR分析结果表明,ZoIPTs响应干旱、淹水、盐胁迫。在淹水和盐胁迫下,根茎中ZoIPT3表达量显著上升；在干旱胁迫下,叶和根茎中ZoIPT5的表达显著上升。综上所述,本研究通过系统的鉴定、进化分析、特征分析、启动子分析、表达模式分析,并对干旱、盐、淹水胁迫下的表达模式进行了分析,为深入研究ZoIPT在调控生姜生长发育和抗逆性中的生物学功能提供了理论基础。
Isopentenyl-transferases (IPT) are key enzymes that catalyze the biosynthesis of cytokinin and are also important rate-limiting enzymes of cytokinin. In plants, IPT exists in the form of ATP/ADP IPTs and TrnA-IPTS, and IPTs mediate the synthesis of two types of CKs in plants: the ATP/ADP pathway and the tRNA pathway. Ginger is an important vegetable that can been used as medicine and food, but it is tend to be affected by various biotic and abiotic stresses during its cultivation, which is detrimental to the safe production of ginger. Here, 10 ZoIPT were identified from ginger genome through systematic bioinformatics analysis, and they were named as ZoIPT1-ZoIPT10. Gene structure analysis showed that all ZoIPTs gene family members contain introns. The amino acid length of the encoded protein ranged from 283 (ZoIPT8) to 491(ZoIPT10) aa, the molecular weight was 31.14 (ZoIPT8) to 54.02 (ZoIPT10) kD, and the isoelectric point was 4.97 (ZoIPT8) to 9.37(ZoIPT6) pI. The analysis of protein characteristics showed that all ZoIPTs were hydrophilic. Subcellular localization analysis showed that 8 ZoIPTs were localized in chloroplasts and 2 ZoIPTs were localized in mitochondria and cytoplasm. Transcriptome data analysis showed that ZoIPTs had tissue specific expression patterns and could respond to stress such as disease and low temperature, ZoIPT3 and ZoIPT5 were highly expressed at different growth stages, different tissues, and in response to low temperature and disease stress. qRT-PCR analysis showed that ZoIPTs was responsive to drought, flooding and salt stress. Under flooding and salt stress, the expression of ZoIPT3 in rhizomes was significantly increased. Under drought stress, the expression of ZoIPT5 in leaves and rhizomes were significantly increased. In summary, through systematic identification, evolutionary analysis, characteristic analysis, promoter analysis, expression pattern analysis, and expression patterns analysis under drought, salt, and flooding stress, this study provided a theoretical basis for in-depth research on the biological functions of ZoIPT in regulating growth and development and stress resistance of ginger.