作物学报2018，Vol.44Issue(4)：512-521,10.DOI:10.3724/SP.J.1006.2018.00512
amiRNA技术沉默C-3氧化酶编码基因StCPD对马铃薯抗旱性的影响
Effect of Silencing C-3 Oxidase Encoded Gene StCPD on Potato Drought Re-sistance by amiRNA Technology
摘要
Abstract
The constitutive photomorphogenesis and dwarf(CPD)gene encodes C-3 oxidase as a key rate-limiting enzyme in the brassinosteroids (BRs) biosynthesis pathway, which plays a vital role in response to abiotic stress in plant. In this research, the potato CPD gene(StCPD)interference expression vector pCPB121-amiRcpd was constructed using the plant expression vector pRS-300 and amiRcpd skeleton precursor of Arabidopsis miR319a,and transferred into the potato genome mediated by Agrobac-tium tumefacienses L.,so that transgenic plants(Ci1–Ci5)were obtained.The analysis of real time fluorescence quantitative po-lymerase chain reaction(qRT-PCR)showed that the interference degree of StCPD gene expression reached 78% and 90% in the transgenic lines Ci1 and Ci3 respectively. StCPD gene expressed in the roots, stems and leaves of the transgenic and non-transgenic (NT) plants, and expression level in the leaves was 3.05 and 1.65 fold higher than that in stems and roots. The malondialdehyde (MDA) content in transgenic plant leaves was significantly higher, whereas the proline content was significantly lower than those in NT under drought stress.The expression level of StCPD gene,MDA and proline contents in transgenic and NT plants under drought stress were significantly higher than those of control, and increased at different sampling times. These results indicated that StCPD gene interference expression could depress the effects of drought stress on potato.These results lay a foun-dation for further research on BRs regulation in potato development, which will help to reveal the molecular mechanisms of BRs regulation in potato.关键词
马铃薯/油菜素内酯/StCPD基因/RNA干扰/干旱胁迫Key words
potato/brassinosteroids/StCPD gene/RNA interference/drought stress引用本文复制引用
周香艳,杨江伟,唐勋,文义凯,张宁,司怀军..amiRNA技术沉默C-3氧化酶编码基因StCPD对马铃薯抗旱性的影响[J].作物学报,2018,44(4):512-521,10.基金项目
本研究由国家自然科学基金项目(31460370,31660416)资助.This study was supported by the National Natural Science Foundation of China(31460370,31660416). （31460370,31660416）
