| Nelumbo nucifera Gaertn.(sacred lotus)is not only one of the important aquatic vegetable crops,but also good material for the research in basic science with extremely resistance to high temperature,accordingly,this includes greater theoretical and practical meaning.The extraction methods of sacred lotus seeds protein were to be selected and optimized using four kinds of protein extraction methods(new TCA/acetone,improved tris-cl,traditional TCA/acetone,tris-saturated phenol methods)combined with two-dimensional electrophoresis(2-DE)technology in the present study.The 2-DE analysis showed that the resulting protein profile/map exhibited consistency with the composition and distribution of the sacred lotus seeds proteome in 'Tielian'.Analysis by PDQuest software showed that the new TCA/acetone method was suitable for 2-DE protein extraction of lotus cotyledons and plantules,whereas the traditional TCA/acetone method was suitable for the 2-DE protein extraction of hypocotyls.This work lays a foundation for further research on the sacred lotus seeds proteome by mass spectrometry.By means of 10-minute heating treatment of different temperature gradient for cotyledon protein samples,then 2-DE to isolate protein and PDQuest software to analyze electrophoretograms,next,the essay can make use of UniprotKB index for protein identification on the basis of MS/MS data combined with HPLC-MS/MS,finally,functions and their roles of thermotolerant proteins in relevant signal control network may work out by means of GO Enrichment analysis and KEGG pathway analysis.Heating treatment and MS experiments showed proteins in Nelumbo fruit that were able to withstand heating stress in cotyledon,proteins that were heat stable at 100 ? and had protective function may be equally important to seed thermostability and viability.Consequently,the results suggested that thermoproteins might be involved in response to heat stress and the process of lotus seed reproduction and development. |
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