五倍体草莓及其十倍体的叶片差异表达蛋白分析

Differentially Expressed Proteins in Strawberry Leaves between Pentaploid and Its Corresponding Allodecaploid

以二倍体绿色草莓（Fragaria viridis Duch.）为父本、八倍体栽培草莓‘房香’（F.×ananassa‘Fusanoka’）为母本杂交所得的五倍体草莓（株系代号：FxLs-11-37,2n=5x=35）及其染色体加倍而成的十倍体（株系代号：A3,2n=10x=70）草莓为试材,观察记载其部分农艺性状、SPAD值以及净光合速率,利用双向凝胶电泳技术对草莓染色体加倍后叶片蛋白质进行了分析,获得了分辨率高、重复性好的电泳图谱。结果表明：（1）与五倍体FxLs-11-37相比,十倍体A3的株型明显矮化、植株冠径变大、叶长叶宽增大、叶片增厚以及叶色浓绿,其叶片的SPAD值与净光合速率显著高于FxLs-11-37。（2）通过PDQuest软件对图谱分析表明,两者在等电点4~7、分子量14.4~66.2kD范围内蛋白质斑点分布最多,可识别的总蛋白质斑点数超过700个,其中蛋白质表达差异水平在1.5倍以上的有18个,2.0倍以上的有4个。利用MALDI-TOF-MS/MS质谱技术鉴定了这4个差异蛋白质,分别是草莓主要过敏原a 1-E、核内不均一核糖核蛋白1、叶绿体硫辛酰基合酶1、NAD（P）H脱氢酶（醌）FQR1类似蛋白质。这些差异蛋白质主要与抗逆、mRNA转运、物质与能量代谢相关。荧光定量PCR对上述4个蛋白编码基因的转录表达水平检测表明,与蛋白质表达差异的趋势一致。该研究获得了草莓染色体加倍后差异表达的蛋白质,为深入研究提供了线索。

Pentaploid strawberry（FxLs-11-37,2n=5x=35）derived from the hybridization between diploid Fragaria viridis（male）and octaploid F.×ananassa‘Fusanoka＇（female）,and its allodecaploid（A3,2n=10x=70）were employed for conducting proteomic analysis with two dimensional gel electrophoresis technology under observation of agronomic traits,soil and plant analyzer development（SPAD）readings,and the net photosynthetic rate.（1）Compared with FxLs-11-37,A3 showed a significant dwarf type,larger crown,increased leaf length and leaf width,much thicker in leaf,more dark green leaves,and it also has a bigger SPAD for leaf and net photosynthetic rate;（2）Protein spots of FxLs-11-37 and A3analyzed using PDQuest software were mainly scattered in the isoelectric point ranging from pH 4to 7,and molecular weight ranging from 14.4to 66.2kD.More than 700 expressed protein spots in FxLs-11-37 and A3were detected.Expression levels of 18 protein spots changed over 1.5fold after chromosome doubling,and4 protein spots changed over 2.0fold.We identified these 4protein spots with MALDI-TOF-MS/MS mass spectrometry technology,and results showed that the 4proteins are major strawberry allergen Fra a 1-E,heterogeneous nuclear ribonucleoprotein 1,lipoyl synthase 1,chloroplastic,and NAD（P）H dehydrogenase（quinine）FQR1-like.These proteins are involved in stress resistant,mRNA transport,material and energy metabolism.Results of real-time quantitative PCR of gene encoding the above four proteins demonstrated that the difference tendency of gene transcriptional expression was consistent with the proteins levels.We obtained the differentially expressed proteins in strawberry chromosome doubling,which provided tracks for further research.