华北农学报
華北農學報
화북농학보
ACTA AGRICULTURAE BOREALI-SINICA
2014年
3期
22-26
,共5页
张玉喜%张文超%李玉娥%刘春英%郑国生%盖树鹏
張玉喜%張文超%李玉娥%劉春英%鄭國生%蓋樹鵬
장옥희%장문초%리옥아%류춘영%정국생%개수붕
牡丹%PsGA20ox基因%内源赤霉素%低温%内休眠解除
牡丹%PsGA20ox基因%內源赤黴素%低溫%內休眠解除
모단%PsGA20ox기인%내원적매소%저온%내휴면해제
Tree peony Paeonia suffroticosa%PsGA20ox%Endogenous gibberellin%Chilling%Endo-dormancy release
为明确在花芽内休眠进程中牡丹赤霉素氧化酶PsGA20ox基因与内源GA3含量的相关性,以454测序筛选到的PsGA20ox基因部分cDNA序列为基础,经RACE扩增和序列拼接后得到1391 bp全长cDNA序列,包括16 bp 5′UTR区,212 bp 3′UTR区,17 bp的PolyA和1146 bp编码区,编码区编码381个氨基酸。同源性分析表明,PsGA20ox与杨树GA20ox同源性最高,为77.6%。表达分析结果表明,在内休眠的前期阶段,PsGA20ox基因的表达水平随着低温处理天数增加而升高,低温处理14 d时PsGA20ox基因表达量最高。酶联免疫反应结果表明,不同低温处理的牡丹花芽内源GA3的含量的变化趋势与PsGA20ox基因表达模式相似。相关分析表明,在鲁荷红休眠解除过程中PsGA20ox基因表达水平与内源GA3含量呈显著正相关。低温累积可诱导PsGA20ox基因的表达,PsGA20ox基因通过调节内源赤霉素含量促进花芽休眠解除。
為明確在花芽內休眠進程中牡丹赤黴素氧化酶PsGA20ox基因與內源GA3含量的相關性,以454測序篩選到的PsGA20ox基因部分cDNA序列為基礎,經RACE擴增和序列拼接後得到1391 bp全長cDNA序列,包括16 bp 5′UTR區,212 bp 3′UTR區,17 bp的PolyA和1146 bp編碼區,編碼區編碼381箇氨基痠。同源性分析錶明,PsGA20ox與楊樹GA20ox同源性最高,為77.6%。錶達分析結果錶明,在內休眠的前期階段,PsGA20ox基因的錶達水平隨著低溫處理天數增加而升高,低溫處理14 d時PsGA20ox基因錶達量最高。酶聯免疫反應結果錶明,不同低溫處理的牡丹花芽內源GA3的含量的變化趨勢與PsGA20ox基因錶達模式相似。相關分析錶明,在魯荷紅休眠解除過程中PsGA20ox基因錶達水平與內源GA3含量呈顯著正相關。低溫纍積可誘導PsGA20ox基因的錶達,PsGA20ox基因通過調節內源赤黴素含量促進花芽休眠解除。
위명학재화아내휴면진정중모단적매소양화매PsGA20ox기인여내원GA3함량적상관성,이454측서사선도적PsGA20ox기인부분cDNA서렬위기출,경RACE확증화서렬병접후득도1391 bp전장cDNA서렬,포괄16 bp 5′UTR구,212 bp 3′UTR구,17 bp적PolyA화1146 bp편마구,편마구편마381개안기산。동원성분석표명,PsGA20ox여양수GA20ox동원성최고,위77.6%。표체분석결과표명,재내휴면적전기계단,PsGA20ox기인적표체수평수착저온처리천수증가이승고,저온처리14 d시PsGA20ox기인표체량최고。매련면역반응결과표명,불동저온처리적모단화아내원GA3적함량적변화추세여PsGA20ox기인표체모식상사。상관분석표명,재로하홍휴면해제과정중PsGA20ox기인표체수평여내원GA3함량정현저정상관。저온루적가유도PsGA20ox기인적표체,PsGA20ox기인통과조절내원적매소함량촉진화아휴면해제。
In order to investigate the correlation between the expression level of PsGA20 ox gene and endoge-nous GA3 contents during dormancy release in tree peony, using tree peony Lu hehong as plant materials, RACE PCR amplification was carried out based on the obtained partial cDNA sequence by 454 sequencing. The results of fragment assembly indicated that full-length cDNA sequence of the PsGA20 ox gene was 1 391 bp including a 1 146 bp open reading frame( ORF) ,16 bp 5′terminal UTR and 212 bp 3′terminal UTR followed by 17 bp polyA tail. A-nalysis of the ORF sequence showed that the ORF of PsGA20ox encoded a protein of 381 amino acids. The result of sequence homology indicated that PsGA20ox protein had relatively highly identity with that of poplar,and there was an identity of 77 . 6%. The results of RT-PCR indicated that the expression of PsGA20 ox continuously increased in the tree peony flower buds during the early stage of dormancy release( from 0 to 14 d chilling treatment) ,followed by gradually decrease until 28 d,and the expression level of all treatment points were more than that of 0 d. The change trend of the endogenous GA3 content by enzyme-linked immunoassay was similar to that of PsGA20ox expres-sion. Significant positive correlation between the expression level of PsGA20ox and the contents of endogenous GA3 was obtained. These results indicated that PsGA20ox played an important role to accelerate dormancy release by reg-ulating the synthesis of endogenous GA3 .
