CAJ | 학술논문

多胺作为一类新型植物生理活性物质,对植物生长发育起着重要作用,在细胞工程与农业生产中具有广阔的应用价值.多胺氧化酶(Polyamine oxidase, PAO)是催化多胺降解的关键酶之一,对调控植物细胞中多胺浓度非常重要,影响植物的生长发育、形态建成过程.根据国际DNA数据库中已报道的PAO蛋白的保守结构域,设计简并引物(degenerate primer),通过RT-PCR方法首次从花生(仲恺花1号)中克隆PAO基因cDNA序列,并利用BLASTn、BLASTp及多重序列排比(Multiple Sequence Alignment)等生物信息学方法对所克隆的cDNA进行序列分析与鉴定,初步确定已从花生中克隆到PAO基因的同系物(homolog).根据克隆的花生PAO cDNA序列(AhPAO1)设计特异引物,通过半定量RT-PCR方法检测花生胚根、胚芽和幼叶中AhPAO1基因的表达,结果发现胚根中AhPAO1基因表达最强,胚芽和幼叶中的表达无明显差异.本研究为进一步了解花生种子萌发和幼苗生长发育过程中多胺氧化酶基因表达的时空特异性奠定实验基础,并对进一步利用植物基因工程技术综合调控花生体内多胺水平有重要实践意义.
다알작위일류신형식물생리활성물질,대식물생장발육기착중요작용,재세포공정여농업생산중구유엄활적응용개치.다알양화매(Polyamine oxidase, PAO)시최화다알강해적관건매지일,대조공식물세포중다알농도비상중요,영향식물적생장발육、형태건성과정.근거국제DNA수거고중이보도적PAO단백적보수결구역,설계간병인물(degenerate primer),통과RT-PCR방법수차종화생(중개화1호)중극륭PAO기인cDNA서렬,병이용BLASTn、BLASTp급다중서렬배비(Multiple Sequence Alignment)등생물신식학방법대소극륭적cDNA진행서렬분석여감정,초보학정이종화생중극륭도PAO기인적동계물(homolog).근거극륭적화생PAO cDNA서렬(AhPAO1)설계특이인물,통과반정량RT-PCR방법검측화생배근、배아화유협중AhPAO1기인적표체,결과발현배근중AhPAO1기인표체최강,배아화유협중적표체무명현차이.본연구위진일보료해화생충자맹발화유묘생장발육과정중다알양화매기인표체적시공특이성전정실험기출,병대진일보이용식물기인공정기술종합조공화생체내다알수평유중요실천의의.
Polyamines are being considered as novel active substances with great significance to plant growth and development and play important potential role in application to cell engineering and agriculture production. Polyamine oxidase (PAO) is one of the key enzymes which catalyze the degradation of polyamines. PAO is of great importance to regulation of polyamines concentration in plant cells and influences growth, development and morphogenesis of plants. In the present study, based on the conserved regions of reported PAO proteins in GenBank database, two degenerate primers were designed to amplify PAO cDNA homolog through reverse transcription polymerase chain reaction (RT-PCR) from peanut (Arachis hypogaea cv Zhongkaihua No. 1). The cloned PAO cDNA, designated as AhPAO1, was characterized and confirmed by BLASTn, BLASTp and Multiple Sequence Alignment analysis bioinformatically, showing that AhPAO1 is the cDNA encoding a putative PAO of peanut plants. Semi-quantitative RT-PCR with two gene-specific primers was performed to investigate the organ specific expression pattern of AhPAO1 gene in radicles, plumules and young leaves of peanut seedlings. The results showed that the expression of AhPAO1 gene in peanut radicles was significantly stronger than that in plumules and young leaves. This investigation provides experimental foundations for further insight into the temporal and spatial expression patterns of AhPAO1 gene during seed germination and seedling development of peanut plants, and for regulation of polyamines level in peanut plants through transgenic plant engineering.