摘要
MAX1(MORE AXILLARY GROWTH1)是独脚金内酯(Strigolactones,SLs)合成途径中的关键基因,为了研究MAX1在苹果分枝调控中的功能,以苹果‘长富2号’(Malus domestica‘Nagafu 2’)腋芽为材料,采用PCR方法,克隆MdMAX1基因,进行生物信息学和表达水平分析;采用瞬时表达转化烟草,进行GUS染色,分析MdMAX1启动子活性。结果表明:(1)成功克隆得到苹果MAX1,其开放阅读框(ORF)1620 bp,编码539个氨基酸;系统进化和基序分析表明,MdMAX1和已知的A1型MAX1相似。(2)qRT-PCR分析表明,MAX1基因在‘长富2号’嫁接苗茎中高表达,并在腋芽本身有表达;RNA-seq分析表明,细胞分裂素(6-BA)处理苹果腋芽96 h后MAX1基因的表达水平显著降低。(3)成功克隆获得‘长富2号’MAX1启动子序列片段(1500 bp),顺式作用元件预测显示MdMAX1启动子序列中存在光响应元件,GUS活性分析表明光照处理能够减弱MAX1启动子的活性。该研究为进一步研究苹果MAX1参与SLs合成、调控苹果分枝的功能奠定了基础。
In order to study the function of MAX1 in the regulation of apple branching,we cloned the MdMAX1 gene by PCR from the axillary buds of Malus domestica‘Nagafu 2’,and carried out the bioinformatics and expression level analysis.We further analyzed the promoter activity of MdMAX1 by GUS staining.The results showed that:(1)apple MAX1 was successfully cloned and its ORF was 1620 bp,encoding 539 amino acids.Phylogenetic analysis and motif analysis showed that MdMAX1 was similar to known A1 type MAX1.(2)qRT-PCR analysis showed that MAX1 was highly expressed in the stems of‘Changfu 2’grafted seedlings,and also in axillary buds.RNA-seq analysis showed that the expression level of MAX1 in apple axillary buds decreased significantly after treated with 6-BA for 96 h.(3)The promoter sequence of‘Changfu 2’MdMAX1(1500 bp)was successfully cloned.The cis acting element prediction showed that there was a light response element in the promoter sequence of MdMAX1.GUS activity analysis showed that the activity of MAX1 promoter was weakened by light treatment.This study provides a basis for further research on the function of MAX1 involved in the synthesis of SLs in regulating apple branches in the future.
作者
高馨怡
陈皓
程方
常兴敏
孙怡婷
谢玲玲
马娟娟
GAO Xinyi;CHEN Hao;CHENG Fang;CHANG Xingmin;SUN Yiting;XIE Lingling;MA Juanjuan(College of Horticulture, Northwest A&F University, Yangling, Shaanxi 712100, China)
出处
《西北植物学报》
CAS
CSCD
北大核心
2021年第1期29-36,共8页
Acta Botanica Boreali-Occidentalia Sinica
基金
农业部国家苹果产业技术体系(CARS-27)
中国博士后项目(2015M582713)。
