| Oil palm (Elaeis guineensis Jacq.),one of the most important oil-producing crops in the tropics, contains large amounts of polyunsaturated fatty acids which has gradually been becoming the hotspot research in the recent years. Palm oil fatty acid content in terms of the type and exhibit canola, soybeans and other bulk oil crops different characteristics, where in the unsaturated fatty acids (oleic and linoleic acids) ratio of about 50% of the total fatty acids.At present, domestic oil palm has not been related to an unsaturated fatty acid molecular mechanism research reports.In this study we were cloned and identified three different oil palm fatty acid desaturase gene (?3, ?6, ?12) promoter sequence, do a study on its regulatory mechanism, achieved the following results:Oil palm leaf genomic DNA was used as templateas, based on the oil palm genome sequence synthesized oil palm desaturase gene ?3, ?6, ?12 promoters pecific primers by PCR amplification and TA cloning method, respectively obtained with a length of 1144 bp, 1474 bp,1233bp oil palm desaturase gene ?3, ?6, ?12 fatty acid desaturase gene promoter sequences.The cis-acting elements of the sequence were analyzed by PlantCARE software,it found that three promoters contain a plant gene promoter specific CAAT-box and TATA box domain;Meanwhile, the promoter sequence cloned also contains a large number of associated light responsive element, and some hormone-responsive elements: such as abscisic acid, ethylene, salicylic acid, methyl jasmonate response components.The gus staining and gus activity expression of transgenic arabidopsis was analyzed by structuring plant expression vector which contains gus and being transformed by Agrobacterium mediated transformation. The result showed that the coined three promoter sequences and downstream of the integration of gus gene has been stable to obtain transgenic arabidopsis genome. In the meanwhile, gene gus showed obvious tissue specificity in different tissues.Among them, the ?3 promoter in the stem and leaf area of tissue specificity, expressing quantity is greatest in the stem, the leaf veins in the expression. A6 promoter in leaf and capsule with tissue specificity,leaf expressing quantity is higher than capsule.?12 promoter in leaf with tissue specificity.In order to study external factors on unsaturated fatty acid regulation of gene expression mechanism, containing ?3, ?6, ?12 promoter transgenic Arabidopsis different photoperiod, temperature, abscisic acid, ethylene, salicylic acid and methyl jasmonate treatment.GUS activity after treatment Quantitative analysis showed that:?3 promoter under the conditions of photoperiod treatment, leaves and stems of GUS activity increased;After the ABA treatment, leaves and stems of GUS activity increased; in the process of JA, leaves GUS protein activity decreased, while the stems GUS protein activity has increased;In the case of 4 "C cold treatment, GUS protein activity leaves and stems showing first increased, then decreased;Under salicylic acid treatment, GUS activity in leaves and stems increased;nder ethephon treatment, GUS protein activity leaves showed an increasing trend, GUS protein activity stems downward trend. ? 6 promoter under the conditions of photoperiod treatment, and the Ministry of pods leaves GUS activity increases; under ABA treatment, GUS protein activity at the blade and pod at significantly increased; in the case of 4? cold treatment next, GUS protein activity leaves and fruit pods increased; JA in the process, leaves and pods of GUS activity decreased.?12 promoter under the conditions according to photoperiod treatment, after GUS protein activity blade at first and then decreased;Abscisic acid, the vane after GUS protein activity at first and then decreased;In the process of JA, GUS protein activity leaves fall; case 4? cold treatment, GUS protein activity leaves decreased; salicylic acid treatment under, GUS activity was significantly increased blade conclusion can be drawn, the promoter of the expression of these three by photoperiod, temperature, abscisic acid, ethylene, salicylic acid and methyl jasmonate hormonal regulation of these. |
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