利用蛋白质组技术分析Cd2+对萝卜幼苗生长的影响

Effects of Cd2+stress on radish (Raphanus sativus) seedling growth based on proteome technique

  • 摘要: 以生长3 d的萝卜幼苗为材料, 分析10~150 mmol·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 mmol·L-1 Cd2+处理, P<0.01)、1.61±0.37 cm (50 mmol·L-1 Cd2+处理, P<0.01)、1.26±0.11 cm (100 mmol·L-1Cd2+ 处理, P<0.01)和0.80±0.14 cm (150 mmol·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。

     

    Abstract: Three-day-old radish seedlings were treated with 10~150 mmol·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 seedling growth. Seedling height decreased from 3.80±0.68 cm under the control to 3.41±0.64 cm under 10 mmol·L-1 Cd2+ treatment (P < 0.01), 1.61±0.37 cm under 50 mmol·L-1Cd2+ treatment (P < 0.01), 1.26±0.11 cm under 100 mmol·L-1 Cd2+ treatment (P < 0.01) and to 0.80±0.14 cm under 150 mmol·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 mmol·L-1, 4.52±0.13 g under 50 mmol·L-1, 3.65±0.07 g under 100 mmol·L-1 and to 1.03±0.01 g under 150 mmol·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, respectively. Proteomic techniques analyses showed that treating seedlings with 100 mmol·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 carboxylase (PEPC), spot-5 (deoxycytidylate deaminase), spot-6 (maturase K), spot-7 (GRAS family transcription factor), spot-8 (resistance protein) and spot-9 puroindoline B (pin). These identified Cd2+ responsive proteins were possibly involved in DNA function 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 adaptation and tolerance of plant to heavy metals.

     

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