CAJ | 학술논문

酸性土壤中的铝毒害是限制作物生长和产量的主要因素之一。拟南芥中的 AtSTOP1(Arabidopsis thaliana sensitive to proton rhizotoxicity 1)是一个调控多种铝毒耐受机制相关基因表达的转录因子,在拟南芥耐铝毒中发挥重要作用。为研究大豆中 STOP1-like 基因的表达特性,本研究利用 RT-PCR 从耐铝毒大豆品种科丰1号中克隆了一个位于第16染色体的 STOP1-like 基因,命名为 GmSTOP1。该基因的编码区(coding DNA sequence, CDS)序列长度为1566 bp,编码521个氨基酸。在 GmSTOP1起始密码子上游1500 bp 的核苷酸序列区间预测到多种顺式作用元件,包括与激素、热、逆境响应等相关的应答元件,如 ABRE、HSE、TC-rich 重复序列等。蛋白质结构预测表明 GmSTOP1不具有跨膜结构和信号肽,含有4个保守的 Cys-2-His-2锌指蛋白结构域。系统进化分析显示 GmSTOP1与菜豆(Phaseolus vulgaris)中的 STOP1-like 蛋白亲缘关系较近。亚细胞定位结果显示 GmSTOP1定位于细胞核,说明GmSTOP1蛋白可能在细胞核中发挥其功能。GmSTOP1基因在种子中的相对表达量最高,在根、茎尖分生组织、茎、叶、花、荚等多种组织中也均有表达。用25μmol L–1 AlCl3溶液处理大豆幼苗, GmSTOP1基因在根中上调表达,24 h达到最高相对表达量,约为对照(0μmol L–1 AlCl3)的9.2倍,表明该基因的表达受铝离子的诱导。此外, ABA、NaCl和 PEG 等胁迫也能诱导大豆根和叶中 GmSTOP1基因的上调表达。由此推测 GmSTOP1基因可能参与大豆对铝毒、高盐和渗透等非生物胁迫的应答过程。
산성토양중적려독해시한제작물생장화산량적주요인소지일。의남개중적 AtSTOP1(Arabidopsis thaliana sensitive to proton rhizotoxicity 1)시일개조공다충려독내수궤제상관기인표체적전록인자,재의남개내려독중발휘중요작용。위연구대두중 STOP1-like 기인적표체특성,본연구이용 RT-PCR 종내려독대두품충과봉1호중극륭료일개위우제16염색체적 STOP1-like 기인,명명위 GmSTOP1。해기인적편마구(coding DNA sequence, CDS)서렬장도위1566 bp,편마521개안기산。재 GmSTOP1기시밀마자상유1500 bp 적핵감산서렬구간예측도다충순식작용원건,포괄여격소、열、역경향응등상관적응답원건,여 ABRE、HSE、TC-rich 중복서렬등。단백질결구예측표명 GmSTOP1불구유과막결구화신호태,함유4개보수적 Cys-2-His-2자지단백결구역。계통진화분석현시 GmSTOP1여채두(Phaseolus vulgaris)중적 STOP1-like 단백친연관계교근。아세포정위결과현시 GmSTOP1정위우세포핵,설명GmSTOP1단백가능재세포핵중발휘기공능。GmSTOP1기인재충자중적상대표체량최고,재근、경첨분생조직、경、협、화、협등다충조직중야균유표체。용25μmol L–1 AlCl3용액처리대두유묘, GmSTOP1기인재근중상조표체,24 h체도최고상대표체량,약위대조(0μmol L–1 AlCl3)적9.2배,표명해기인적표체수려리자적유도。차외, ABA、NaCl화 PEG 등협박야능유도대두근화협중 GmSTOP1기인적상조표체。유차추측 GmSTOP1기인가능삼여대두대려독、고염화삼투등비생물협박적응답과정。
Aluminum toxicity is one of the major factors that limits the growth and production of crops in acid soils. AtSTOP1 transcription factor can regulate the expression of genes related to aluminum-toxicity tolerance mechanisms, which plays an im-portant role in aluminum-toxicity tolerance in Arabidopsis. To study the expression features of the STOP1-like gene in soybean, we cloned a STOP1 gene located on chromosome 16 from the aluminum-toxicity tolerant soybean cultivar (Kefeng-1) using RT-PCR, and designated as GmSTOP1. The length of GmSTOP1 coding DNA sequence was 1566 bp, which encoded 521 amino acid residues. Diverse cis-acting promoter elements involved in hormone, heat and stress responses were discovered in the 1500 bp upstream region of GmSTOP1, such as ABRE, HSE, TC-rich repeats, and other elements. Protein structure prediction showed that it did not have any signal-peptide or transmembrane region, but contained four conservative Cys-2-His-2 zinc-finger domains. Phylogenetic analysis demonstrated that GmSTOP1 was similar to the putative STOP1-like protein from Phaseolus vulgaris. Re-sults of subcellular localization showed that GmSTOP1 protein is located in the cell nucleus. The transcripts of GmSTOP1 were detected in all organs tested including root, shoot apical meristem, stem, leaf, flower, pod and seed, with the highest level in seed. GmSTOP1 was up-regulated in soybean roots by 25 μmol L–1 AlCl3 treatment, and reached the highest relative expression level at 24 hours, which was about 9.2 times of the level in control (0 μmol L–1 AlCl3). In addition, Real-time PCR analysis showed that the expression of GmSTOP1 in soybean leaf and root was also up-regulated by ABA, NaCl, and PEG, respectively. These results indicated that GmSTOP1 might participate in soybean response to abiotic stresses including aluminum-toxicity, high salinity and osmosis stress, which provides the basis for further studying the functions of GmSTOP1.