利用蛋白质组技术分析Cd^(2＋)对萝卜幼苗生长的影响  

Effects of Cd^(2+) stress on radish(Raphanus sativus) seedling growth based on proteome technique

作　　者：党晨[1] 高越[2] 阎晗[2] 彭永康[2] 

机构地区：[1]中国农业大学农学与生物技术学院,北京100193 [2]天津师范大学生命科学学院,天津300387

出　　处：《中国生态农业学报》2012年第2期231-235,共5页Chinese Journal of Eco-Agriculture

基　　金：教育部科学技术研究重点项目(02010)资助

摘　　要：以生长3 d的萝卜幼苗为材料,分析10~150?mol.L?1 Cd2+处理12 h后幼苗的生长及蛋白质组的变化。结果表明,不同浓度Cd2+处理导致幼苗生长严重受抑,幼苗高度从对照组的3.80?0.68 cm降至3.41?0.64 cm(10?mol.L?1 Cd2+处理,P<0.01)、1.61±0.37 cm(50?mol.L?1 Cd2+处理,P<0.01)、1.26±0.11 cm(100?mol.L?1 Cd2+处理,P<0.01)和0.80±0.14 cm(150?mol.L?1 Cd2+处理,P<0.01);胚根生长也明显受到影响,分生组织细胞有丝分裂受抑,幼苗鲜重下降;叶绿素(a+b)含量[mg.g?1(FW)]从对照组的6.72±0.05分别下降至6.66±0.17、6.02±0.15、5.38±0.07和3.94±0.06。蛋白质组技术分析表明,叶片中有50多个蛋白质斑点产生差异表达现象,其中9个蛋白质斑点的归属得以鉴别,分别是斑点1 PWWP domain containing protein、斑点2 AAA-type ATPase family protein、斑点3 NB-ARC domain containing protein、斑点4 Phosphoenolpyruvate carboxylase(PEPC)、斑点5 Deoxycytidylate deaminase、斑点6 Maturase K、斑点7 GRAS family transcription factor、斑点8 Resistance protein和斑点9 Puroindoline B(pin),其功能涉及DNA功能修饰(甲基化)、能量代谢、信号转导、蛋白质合成、基因转座与剪切和不良环境条件防御等。Three-day-old radish seedlings were treated with 10～150 μmol·L-1 Cd2＋ for 12 h for determination of the effects of Cd2＋ on seedling growth and proteome by using 2-DE technique and MALDI-TOF MS.The results showed an obvious inhibition of seed-ling growth.Seedling height decreased from 3.80±0.68 cm under the control to 3.41±0.64 cm under 10 μmol·L-1 Cd2＋ treatment（P 0.01）,1.61±0.37 cm under 50 μmol·L-1 Cd2＋ treatment（P 0.01）,1.26±0.11 cm under 100 μmol·L-1 Cd2＋ treatment（P 0.01） and to 0.80±0.14 cm under 150 μmol·L-1 Cd2＋ treatment（P 0.01）.Root growth was also obviously inhibited.Cell mitotic inhibition was noted in root tip meristem.Seedling fresh weight decreased from 10.92±0.86 g under the control to 9.93±0.77 g under 10 μmol·L-1,4.52±0.13 g under 50 μmol·L-1,3.65±0.07 g under 100 μmol·L-1 and to 1.03±0.01 g under 150 μmol·L-1 Cd2＋ treatments.Similarly,chlorophyll（a＋b） content [mg-g-1（FW）] declined from 6.72±0.05 to 6.66±0.17,6.02±0.15,5.38±0.07 and 3.94±0.06,re-spectively.Proteomic techniques analyses showed that treating seedlings with 100 μmol·L-1 Cd2＋ altered over 50 protein species.9 protein spots were identified via the MS/MS approach.The identified protein sports included： spot-1（PWWP domain containing protein）,spot-2（AAA-type ATPase family protein）,spot-3（NB-ARC domain containing protein）,spot-4 [phosphoenolpyruvate car-boxylase（PEPC）],spot-5（deoxycytidylate deaminase）,spot-6（maturase K）,spot-7（GRAS family transcription factor）,spot-8（re-sistance protein） and spot-9 [puroindoline B（pin）].These identified Cd2＋ responsive proteins were possibly involved in DNA func-tion modification（DNA methylation）,energy metabolism,cell signal transduction,protein biosynthesis,gene transposition,intron splicing and defense response.This suggested that several protein types were responsive to Cd2＋ stress.Proteome technique was applicable in studying physiological and biochemical mechanisms of adapta

关 键 词：萝卜 CD2+胁迫 叶绿素(a+b)含量 蛋白质组技术 串联质谱 幼苗生长 

分 类 号：Q89[生物学]