摘要
为探讨茭白冷藏期间衰老的分子机理,应用蛋白质组学技术,研究了茭白冷藏期间蛋白质表达谱的变化。结果显示,双向电泳胶上共检测到大约650个蛋白点,其中35个蛋白表达量存在2.0倍以上显著(p<0.05)差异,经过串联飞行时间质谱分析,成功鉴定出29个蛋白,根据其功能可分为6类,即代谢(20.7%)、细胞结构(27.6%)、抗胁迫(20.7%)、衰老(6.9%)、蛋白质合成(13.8%)和功能未知蛋白(10.3%);其中:代谢相关蛋白3个上调表达、3个下调表达,细胞结构相关蛋白6个上调表达、2个下调表达,抗胁迫相关蛋白4个上调表达、2个下调表达,衰老相关蛋白2个上调表达,蛋白质合成相关蛋白4个及功能未知蛋白3个均下调表达。这些差异表达蛋白的功能分析表明,茭白采后衰老机理可能涉及物质代谢过程的调整、能量代谢途径的改变、活性氧清除能力的减弱以及细胞结构的解体。
The changes of protein profile in postharvest Z. latifolia during cooling storage at 1°C for 0 （control）, 15 and 30 days were investigated to study the molecular mechanism of senescence of postharvest Zizania latifoliausing two-dimensional electrophoresis （2-DE）. The results showed that a total of about 650 protein spots were detected on the gels in which 35 spots showed a significant （p〈0.05） change in protein abundance based on the two-fold difference. 29 spots were identified using MALDI-TOF/TOF. We classified these 29 protein spots into six functional categories： metabolism （20.7%）, cell structure （27.6%）, stress response and defense （20.7%）, senescence （6.9%）, protein synthesis （13.8%） and unknown functional proteins （10.3%）. 15 spots were up-regulated while 14 spots were down-regulated among 29 differentially expressed proteins spots. For 15 up-regulated spots, spot 9 （adenosine kinase）, spot 23 （glycosyl hydrolases family 17） and spot 27 （transketolase） were related to metabolism; spot 1 （profiling）, spot 6 （β-1,3-glucanase precursor）, spot 7 （glucan endo-1,3-beta-glucosidase）, spot 8 （β-1,3-glucanase precursor）, spot 24 （xyloglucan endotransglycosylases） and spot 39 （β-1,3-glucanase） were related to cell structure; spot 17 （universal stress protein family）, spot 30 （thaumatin-like protein isoform 2）, spot 32 （glutathione S-transferase） and spot 48 （dehydroascorbate reductase） were related to stress response and defense; spot 22 （cysteine protease） and spot 33 （papain-like cysteine proteinase） were related to senescence. For 14 down-regulated spots, spot 36 （diphosphonucleotide phosphatase 1）, spot 44 （enolase） and spot 53 （isocitrate dehydrogenase） were related to metabolism;spot 25 （actin） and spot 38 （actin depolymerisation factor/cofilin -like domains） were related to cell structure; spot 31 （hipI-SODC1s） and spot 49 （chitinase class I） were related to stress response and defense;spot 15 （glycine-rich RNA-binding protein）, spot 18 （putative chaperonin 21 precursor）, spot 21 （putative ribosomal protein S12） and spot 56 （ubiquitin-conjugating enzyme E2） were related to protein synthesis;spot 43 （WD40 domain）, spot 51 （unknown protein） and spot 55 （band_7_prohibitin） were unknown functional proteins. The most important proteins that behave differentially are related to senescence in postharvest Z. latifolia wereβ-1,3-glucanase precursor （spot 6, spot 8）, adenosine kinase （spot 9）, universal stress protein family （spot 17）, cysteine protease （spot 22）, glycosyl hydrolases family 17 （spot 23）, hipI-SODC1s （spot 31）, glutathione S-transferase （spot 32）, papain-like cysteine proteinase （spot 33）,β-1,3-glucanase （spot 39）, dehydroascorbate reductase （spot 48） and chitinase class I （spot 49）. These protein’s functions mainly concentrated on material and energy metabolism, stress response and defense, cell structure and senescence. These results indicated that the senescence of postharvest Z. latifolia may be caused by multi-actions including the regulation of material metabolism, change of energy metabolism pathway, decline of reactive oxygen scavenging capacity and degradation of cell structure. This study provided experimental data and come up with new perspectives for the mechanism of postharvest Z. latifolia senescence. However, the molecular biological analysis and subcellular proteomic studies need to be further investigated in the future.
出处
《农业工程学报》
EI
CAS
CSCD
北大核心
2014年第1期285-292,共8页
Transactions of the Chinese Society of Agricultural Engineering
基金
"十二五"国家科技支撑计划项目(2012BAD27B03)
关键词
蛋白质
冷藏
电泳
质谱
衰老
茭白
proteins
cold storage
electrophoresis
mass spectrometry
slow death
Zizania latifolia
