
盐胁迫下三倍体小黑杨杂种无性系叶片蛋白质差异表达分析
张恒, 刘晓婷, 陈嵩, 周雪燕, 司冬晶, 李莹, 赵曦阳
南京林业大学学报(自然科学版) ›› 2020, Vol. 44 ›› Issue (2) : 59-66.
盐胁迫下三倍体小黑杨杂种无性系叶片蛋白质差异表达分析
Analysis of differentially expressed proteins in leaves of triploid Populus simonii × P. nigra hybrid clones under salt stress
【目的】探究三倍体小黑杨生长迅速原因。通过蛋白质差异表达分析,深入研究三倍体小黑杨生长发育机理,为其优质高产育种提供参考。【方法】以三倍体小黑杨杂种无性系为材料,分别用75 mmol/L NaCl溶液胁迫处理0、4、8、12和16 d,利用蛋白质双向电泳和MALDI-TOF/TOF质谱技术,探讨其不同处理条件下叶片差异表达蛋白。【结果】共检测到4 000多个蛋白点,包含96个差异蛋白点,其中盐胁迫4、8、12、16 d与对照相比,分别鉴定出5、21、36、34个差异蛋白点,对60个蛋白点进行质谱鉴定,这些蛋白按功能可分为8类:其中光合作用占25%、能量代谢占20%、氧化还原作用占12%、抗性蛋白占8%、生物合成和代谢占7%、运输蛋白占3%、信号转导占2%和未知蛋白占23%。【结论】三倍体小黑杨杂种无性系受到盐胁迫后,多种蛋白质共同参与应答,其中主要是光合作用和能量代谢相关蛋白,本研究可以为盐胁迫下林木蛋白质组学研究提供基础。
【Objective】 Polyploid breeding has widely been used in the genetic improvement of different tree species because of the rapid and extensive growth of polyploid plants. There are many studies describing polyploidy in Populus hybrids, but most of them focused on growth traits, wood properties and stress tolerance, and few studied their growth and development mechanisms. In this study, the salt tolerance mechanisms of polyploid were deeply studied through proteomics analysis, and their metabolic regulation mechanisms were revealed, which could provide a theoretical reference for their high yield breeding. We studied salt tolerance mechanisms of triploid Populus hybrids, which was of great significance for improving salt-tolerance of P. simonii × P. nigra. 【Method】 In this study, we used triploidP. simonii × P. nigra hybrid clones which were treated with 75 mmol/L NaCl solution for 0, 4, 8, 12 and 16 days, and we analyzed the differentially expressed proteins from leaves by two-dimensional electrophoresis and MALDI-TOF/TOF mass spectrometry under salt stress in different treatment conditions. 【Result】 More than 4 000 protein spots were detected in this study. There were 96 protein spots showing different expression patterns after the treatment. We identified 5, 21, 36 and 34 differentially expressed protein spots under salt stress on the 4th, 8th, 12th and 16th day, respectively, compared with the control group. The up-regulated proteins were 1, 7, 17 and 9, respectively, while the rest were down-regulated proteins. Sixty proteins were successfully identified by mass spectrometry. These proteins could be divided into eight categories according to their functions, and 15 photosynthesis-related proteins, including phosphoribulokinase and photosystem Ⅱ oxygen-evolving complex protein two precursor, accounted for 25% of the total protein content. There were 12 energy metabolism-related proteins, including triosephosphate isomerase and fructose-bisphosphate aldolase 1, which accounted for 20% of the total protein content. SOD, glyoxalase I homolog family protein, and seven other redox-related proteins accounted for 12% of the total protein content. H+-transporting two-sector ATPase family of proteins and five other resistant proteins accounted for 8% of the total protein content. Four biosynthesis and metabolism proteins, including S-adenosylmethionine synthase 3, accounted for 7% of the total protein content. Two transport proteins accounted for 3% of the total protein content, one belonging to the MD-2-related lipid recognition domain-containing family of proteins, and the other being a temperature-induced lipocalin. The signal transduction protein only contained thylakoid lumeinal 29 ku protein and accounted for 2% of the total protein content, and 14 unknown proteins accounted for 23% of the total protein content. 【Conclusion】 Under salt stress, multiple proteins (mainly photosynthesis and energy metabolism related proteins) were co-involved in the response to salt stress. This study can provide a basis for future studies of tree proteomics under salt stress.
三倍体小黑杨 / 盐胁迫 / 差异蛋白 / 蛋白质双向电泳 / MALDI-TOF/TOF质谱
triploid Populus simonii × P. nigra / salt stress / differential protein / two-dimensional electrophoresis / MALDI-TOF/TOF mass spectrometry
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