| Salt damage is one of the most important abiotic stresses in global agricultural regions. There are 20% of global land area and over 50% of agricultural irrigated land influenced by salt damages,how to cope with soil salinization has become one of research hotspots nowadays. Industrial hemp is one of the important natural green fiber crops, which has the characteristics of wide distribution, deep root system, fast growth, high biomass, strong resistance to stress and ease cultivation. It can adapt to the stress environment of saline alkali land. The research of salt-tolerant of hemp mainly get through cultivation and physiological way, but the molecular mechanisms of adaption to salt stress remain unclear. In this study,We used salt-tolerant (BM) and salt-sensitive(YM) industrial hemps as experimental materials, established a protein sample preparation method and studied the comparative proteome analysis of hemp under salt stress. The main results were as following:1. This article establishes a protein extected system that is suitable for the proteomic analysis of hemp leaf after optimizing the traditional method:Protein lysis buffer (5 M urea,2M thiourea,1% Triton X-100,50mM Tris-Hcl(PH 8.8),1%DTT),4 times volume of acetone added after protein purification; The number of protein in the leaves were 1027±12 by two-dimensional electrophoret,and the quality of protein can meet requirement of next iTRAQ proteomics.2. The soluble sugar, soluble protein content and SOD content in salt-sensitive YM rapidly decreased in 2d after salt stress;and chlorophyll content continuously decreased with salt stress, with an increased rate of descent after 2d.The contents of Fv/Fm, ETR, qP in two varieties have a downward trend on the whole under salt stress, but qN is on the rise. Physiological datas indicated salt-sensitive variety YM was most sensitive to stress at the 4th day of salt stress, therefore the differential proteomics analysis was conducted at the 4th day of each treatment in this experiment.3. Through study on differential proteomics in the leaves of control, salt-treated BM and YM at 4d of salt stress,3799 proteins were identified, and 3599 proteins were successfully matched by BLAST (NCBI). The result demonstrated that 34.5% of the matched protein annotation species were from the Rosaceae, with 403 and 252 differential proteins in BM and YM, respectively. These differential proteins could be classified into 14 categories:primary metabolism, energy, orientation and storage of protein, disease/defense, protein synthesis, photosynthesis, transport, transcription, signal transduction, cell structure, secondary metabolism, intracellular transport, cell growth/differentiation and unknown.4. Hemp may adapt to stress mainly through improve ATP metabolism, regulate the photosynthesis according to light intensity, strengthen chlorophyll synthesis, promote cell relaxation and enlargement, improve the synthesis of osmoregulation substances, enhanced the flow of inorganic sulfur in body, regulate aquaporins, strengthen the transmission of ion transport signals, improve the signal transmission between proteins, as well as between proteins and cytomembranes, improve the selective absorption and transportation speed of organic and inorganic molecular, degrade hemicellulose cell wall, control cellular material in and out, promote metabolism and cell stability. The test results can provide important reference information for the researches on molecular mechanism of salt-stress adaption in hemp. |
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