植物资源与环境学报
植物資源與環境學報
식물자원여배경학보
JOURNAL OF PLANT RESOURCES AND ENVIRONMENT
2014年
2期
1-10
,共10页
田畅%蒋倩%王枫%李梦瑶%贾晓玲%熊爱生
田暢%蔣倩%王楓%李夢瑤%賈曉玲%熊愛生
전창%장천%왕풍%리몽요%가효령%웅애생
芹菜%Agnp-G3PDH基因%序列分析%同源性%非生物胁迫%表达特性
芹菜%Agnp-G3PDH基因%序列分析%同源性%非生物脅迫%錶達特性
근채%Agnp-G3PDH기인%서렬분석%동원성%비생물협박%표체특성
Apium graveolens Linn.%Agnp-G3PDH gene%sequence analysis%homology%abiotic stress%expression characteristics
从芹菜( Apium graveolens Linn.)品种‘六合黄心芹’(‘Liuhe Huangxinqin’)、‘津南实芹’(‘Jinnan Shiqin’)和‘文图拉’(‘Ventura’)中分别克隆获得非磷酸化甘油醛-3-磷酸脱氢酶基因 Agnp-G3PDH。3个品种的 Agnp-G3PDH基因序列均包含1个全长1491 bp的开放阅读框(ORF),编码496个氨基酸；存在84个碱基位点差异,导致14个氨基酸位点改变。由该基因编码的‘六合黄心芹’、‘津南实芹’和‘文图拉’的Agnp-G3PDH蛋白的理论相对分子质量分别为53201.6、53051.4和52960.3,理论等电点分别为pI 7.49、pI 7.86和pI 8.12,氨基酸组成中碱性氨基酸数量大于酸性氨基酸数量；该蛋白质具有亲水性与疏水性双重特性,均为疏水性蛋白。多重比对结果表明：3个品种Agnp-G3PDH基因编码的氨基酸序列同源性达到99.09％,具有高度保守性；且与其他11种植物np-G3PDH的氨基酸序列的相似度也较高。在基于np-G3PDH基因编码的氨基酸序列进化树上,3个芹菜品种聚在同一分支中,并与杨柳科(Salicaceae)种类毛果杨(Populus trichocarpa Torr.& Gray)聚在一起,表明它们的np-G3PDH进化关系较近。实时定量PCR分析结果表明：在3个芹菜品种的不同组织中Agnp-G3PDH基因的相对表达水平均有明显差异,其中,在‘津南实芹’的叶、‘文图拉’的花和‘六合黄心芹’的根中该基因的相对表达水平最高；经高温(38℃)、低温(4℃)和干旱(20％PEG)胁迫处理后3个品种Agnp-G3PDH基因的相对表达水平均极显著高于或低于对照,但经盐(0.2 mol·L-1 NaCl)胁迫处理后仅品种‘津南实芹’ Agnp-G3PDH基因的相对表达水平显著高于对照,品种‘六合黄心芹’和‘文图拉’Agnp-G3PDH基因的相对表达水平与对照无显著差异。表明该基因的表达具有组织特异性且与品种间的抗逆性差异有关。
從芹菜( Apium graveolens Linn.)品種‘六閤黃心芹’(‘Liuhe Huangxinqin’)、‘津南實芹’(‘Jinnan Shiqin’)和‘文圖拉’(‘Ventura’)中分彆剋隆穫得非燐痠化甘油醛-3-燐痠脫氫酶基因 Agnp-G3PDH。3箇品種的 Agnp-G3PDH基因序列均包含1箇全長1491 bp的開放閱讀框(ORF),編碼496箇氨基痠；存在84箇堿基位點差異,導緻14箇氨基痠位點改變。由該基因編碼的‘六閤黃心芹’、‘津南實芹’和‘文圖拉’的Agnp-G3PDH蛋白的理論相對分子質量分彆為53201.6、53051.4和52960.3,理論等電點分彆為pI 7.49、pI 7.86和pI 8.12,氨基痠組成中堿性氨基痠數量大于痠性氨基痠數量；該蛋白質具有親水性與疏水性雙重特性,均為疏水性蛋白。多重比對結果錶明：3箇品種Agnp-G3PDH基因編碼的氨基痠序列同源性達到99.09％,具有高度保守性；且與其他11種植物np-G3PDH的氨基痠序列的相似度也較高。在基于np-G3PDH基因編碼的氨基痠序列進化樹上,3箇芹菜品種聚在同一分支中,併與楊柳科(Salicaceae)種類毛果楊(Populus trichocarpa Torr.& Gray)聚在一起,錶明它們的np-G3PDH進化關繫較近。實時定量PCR分析結果錶明：在3箇芹菜品種的不同組織中Agnp-G3PDH基因的相對錶達水平均有明顯差異,其中,在‘津南實芹’的葉、‘文圖拉’的花和‘六閤黃心芹’的根中該基因的相對錶達水平最高；經高溫(38℃)、低溫(4℃)和榦旱(20％PEG)脅迫處理後3箇品種Agnp-G3PDH基因的相對錶達水平均極顯著高于或低于對照,但經鹽(0.2 mol·L-1 NaCl)脅迫處理後僅品種‘津南實芹’ Agnp-G3PDH基因的相對錶達水平顯著高于對照,品種‘六閤黃心芹’和‘文圖拉’Agnp-G3PDH基因的相對錶達水平與對照無顯著差異。錶明該基因的錶達具有組織特異性且與品種間的抗逆性差異有關。
종근채( Apium graveolens Linn.)품충‘륙합황심근’(‘Liuhe Huangxinqin’)、‘진남실근’(‘Jinnan Shiqin’)화‘문도랍’(‘Ventura’)중분별극륭획득비린산화감유철-3-린산탈경매기인 Agnp-G3PDH。3개품충적 Agnp-G3PDH기인서렬균포함1개전장1491 bp적개방열독광(ORF),편마496개안기산；존재84개감기위점차이,도치14개안기산위점개변。유해기인편마적‘륙합황심근’、‘진남실근’화‘문도랍’적Agnp-G3PDH단백적이론상대분자질량분별위53201.6、53051.4화52960.3,이론등전점분별위pI 7.49、pI 7.86화pI 8.12,안기산조성중감성안기산수량대우산성안기산수량；해단백질구유친수성여소수성쌍중특성,균위소수성단백。다중비대결과표명：3개품충Agnp-G3PDH기인편마적안기산서렬동원성체도99.09％,구유고도보수성；차여기타11충식물np-G3PDH적안기산서렬적상사도야교고。재기우np-G3PDH기인편마적안기산서렬진화수상,3개근채품충취재동일분지중,병여양류과(Salicaceae)충류모과양(Populus trichocarpa Torr.& Gray)취재일기,표명타문적np-G3PDH진화관계교근。실시정량PCR분석결과표명：재3개근채품충적불동조직중Agnp-G3PDH기인적상대표체수평균유명현차이,기중,재‘진남실근’적협、‘문도랍’적화화‘륙합황심근’적근중해기인적상대표체수평최고；경고온(38℃)、저온(4℃)화간한(20％PEG)협박처리후3개품충Agnp-G3PDH기인적상대표체수평균겁현저고우혹저우대조,단경염(0.2 mol·L-1 NaCl)협박처리후부품충‘진남실근’ Agnp-G3PDH기인적상대표체수평현저고우대조,품충‘륙합황심근’화‘문도랍’Agnp-G3PDH기인적상대표체수평여대조무현저차이。표명해기인적표체구유조직특이성차여품충간적항역성차이유관。
Non-phosphorylating glyceraldehyde-3-phosphate dehydrogenase gene Agnp-G3 PDH was cloned from cultivars ‘Liuhe Huangxinqin’ , ‘Jinnan Shiqin’ and ‘Ventura’ of Apium graveolens Linn., respectively. The sequence of Agnp-G3PDH gene from three cultivars contains an open reading frame ( ORF) with the full length of 1 491 bp, which encodes 496 amino acids. There are 84 base site differences, leading to site changes of 14 amino acids. Theoretical relative molecular mass of protein encoded by Agnp-G3 PDH gene from‘Liuhe Huangxinqin’ ,‘Jinnan Shiqin’ and‘Ventura’ is 53 201 . 6 , 53 051. 4 and 52 960. 3, respectively, and theoretical isoelectric point is pI 7. 49, pI 7. 86 and pI 8. 12,respectively. The number of basic amino acids is more than that of acidic amino acids in amino acid composition. And the proteins all possess double characteristics of hydrophilicity and hydrophobicity and all are the hydrophobic protein. The alignment result shows that homology of amino acid sequences encoded by Agnp-G3PDH gene from three cultivars reaches 99. 09%, which is highly conserved. Similarity of amino acid sequence among three cultivars and other eleven species is higher. Based on phylogenetic tree of amino acid sequence encoded by np-G3PDH gene, three cultivars are clustered in the same branch, and three cultivars and Populus trichocarpa Torr. & Gray in Salicaceae can be clustered together which shows that their np-G3PDH amino acid sequences have a closer evolutionary relationship. The result of quantitative real-time PCR shows that there are obvious differences in relative expression level of Agnp-G3PDH gene among different tissues of three cultivars, in which, relative expression level of the gene is the highest in leaf of ‘Jinnan Shiqin ’ , in flower of ‘Ventura ’ and in root of ‘Liuhe Huangxinqin’ . After treated by stress of high temperature (38℃) , low temperature (4℃) and drought (20%PEG), relative expression level of Agnp-G3PDH gene from three cultivars is extremely significantly higher or lower than that of the control, but after treated by salinity (0. 2 mol·L-1 NaCl) stress only that of ‘Jinnan Shiqin’ is significantly higher than that of the control, and that of ‘Liuhe Huangxinqin’ and‘Ventura’ has no significant difference with that of the control. It is indicated that the expression of Agnp-G3PDH gene has tissue specificity and is related to resistance difference among cultivars.
