华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2014年
6期
21-27
,共7页
田义轲%白牡丹%王彩虹%刘云龙%陈宝印
田義軻%白牡丹%王綵虹%劉雲龍%陳寶印
전의가%백모단%왕채홍%류운룡%진보인
苹果%赤霉素%柯巴焦磷酸合酶基因(CPS)%柱型性状
蘋果%赤黴素%柯巴焦燐痠閤酶基因(CPS)%柱型性狀
평과%적매소%가파초린산합매기인(CPS)%주형성상
Apple%Gibberellins%Ent-copalyl diphosphate synthase(CPS)%Columnar trait
为研究赤霉素合成途径关键酶基因,了解果树的矮化机理。以苹果品种富士当年生新梢的茎尖为试材,以苹果基因组数据库为依据,克隆了编码苹果CPS的基因MdCPS(GenBanK登录号:KC433942.1)。该基因gDNA序列含有15个外显子和14个内含子,其编码序列( Coding sequence,CDS)长度为2400 bp,共编码799个氨基酸。在已发表的金冠苹果基因组中,与该基因相对应的转录本是MDP0000147908,它的定位区间为chr11:32433834~32439214。同源性分析表明,MdCPS与其他植物的CPS间有较高的相似性(49%~67%)。以杂交组合富士(普通型)×舞姿(柱型)的亲本品种及F1后代当年生新梢的茎尖组织为试材,实时荧光定量PCR( qRT-PCR)分析表明,尽管该基因在柱型亲本中的表达水平显著低于普通型亲本,但在其后代的柱型与普通型群体间差异并不明显。同时,在柱型杂种及其普通型突变体间的分析结果也表明,MdCPS的表达水平与柱型性状没有明显的相关性。可见,柱型苹果茎尖组织中活性赤霉素含量偏低受其合成早期步骤关键酶基因MdCPS的影响不大。
為研究赤黴素閤成途徑關鍵酶基因,瞭解果樹的矮化機理。以蘋果品種富士噹年生新梢的莖尖為試材,以蘋果基因組數據庫為依據,剋隆瞭編碼蘋果CPS的基因MdCPS(GenBanK登錄號:KC433942.1)。該基因gDNA序列含有15箇外顯子和14箇內含子,其編碼序列( Coding sequence,CDS)長度為2400 bp,共編碼799箇氨基痠。在已髮錶的金冠蘋果基因組中,與該基因相對應的轉錄本是MDP0000147908,它的定位區間為chr11:32433834~32439214。同源性分析錶明,MdCPS與其他植物的CPS間有較高的相似性(49%~67%)。以雜交組閤富士(普通型)×舞姿(柱型)的親本品種及F1後代噹年生新梢的莖尖組織為試材,實時熒光定量PCR( qRT-PCR)分析錶明,儘管該基因在柱型親本中的錶達水平顯著低于普通型親本,但在其後代的柱型與普通型群體間差異併不明顯。同時,在柱型雜種及其普通型突變體間的分析結果也錶明,MdCPS的錶達水平與柱型性狀沒有明顯的相關性。可見,柱型蘋果莖尖組織中活性赤黴素含量偏低受其閤成早期步驟關鍵酶基因MdCPS的影響不大。
위연구적매소합성도경관건매기인,료해과수적왜화궤리。이평과품충부사당년생신소적경첨위시재,이평과기인조수거고위의거,극륭료편마평과CPS적기인MdCPS(GenBanK등록호:KC433942.1)。해기인gDNA서렬함유15개외현자화14개내함자,기편마서렬( Coding sequence,CDS)장도위2400 bp,공편마799개안기산。재이발표적금관평과기인조중,여해기인상대응적전록본시MDP0000147908,타적정위구간위chr11:32433834~32439214。동원성분석표명,MdCPS여기타식물적CPS간유교고적상사성(49%~67%)。이잡교조합부사(보통형)×무자(주형)적친본품충급F1후대당년생신소적경첨조직위시재,실시형광정량PCR( qRT-PCR)분석표명,진관해기인재주형친본중적표체수평현저저우보통형친본,단재기후대적주형여보통형군체간차이병불명현。동시,재주형잡충급기보통형돌변체간적분석결과야표명,MdCPS적표체수평여주형성상몰유명현적상관성。가견,주형평과경첨조직중활성적매소함량편저수기합성조기보취관건매기인MdCPS적영향불대。
Gibberellines are the most important phtyohormones influencing the plant height of fruit trees. Re-searches of the genes encoding the critical enzymes in gibberellin biosynthesis are significant to understanding the mechanism of dwarf tree architectures. This study reported the isolation of the MdCPS ( GenBank accession number:KC433942. 1),encoding CPS in apple, from primary apical shoots of the variety Fuji based on the apple genome database. The gDNA sequence of MdCPS contained 15 exons and 14 introns,and the coding sequence( CDS) of it was 2 400 bp,which encoded a polypeptide of 799 amino acids. The transcript MDP0000147908 that spanned chro-mosome 11 from location 32433834 to 32439214 in the published Golden delicious apple genome corresponds to this gene. Homology analysis indicated that the deduced MdCPS shared a higher level of similarity ( 49%-67%) with CPS protein from other plant species. Using the primary apical shoots of the parents and F1 progenies of Fuji( stand-ard) × Telamon ( columnar ) as plant materials, quantitative real-time PCR ( qRT-PCR ) analyses showed that even though the transcription level of this gene in the columnar parent was lower than that in the standard parent,there was no significant difference was observed between the two populations of columnar and standard progenies. At the same time,qRT-PCR analysis in the columnar hybrids and their correspondent standard mutants indicated that the expression level of MdCPS was not relevant to the columnar growth habit. These results suggested that the lower con-tent of active gibberellins in columnar apple apical shoots was not distinctively influenced by MdCPS in the early stage of gibberellins synthesis.