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摘要：以花青苷敏感型的‘杨花’萝卜芽苗为试验材料，合适的光照培养条件下在营养液中外源添加不同浓度的硫化氢（H2S）供体硫氢化钠（NaHS）作为处理，以不加硫氢化钠的营养液为对照，探究硫化氢对花青苷合成的影响。实验结果表明，在一定硫氢化钠浓度范围内（0.5 mM～2 mM），随着硫氢化钠浓度的升高，花青苷含量显著提高。对结构基因和转录因子表达量的测定结果表明，硫氢化钠处理后，花青苷合成途径中的关键结构基因PAL、CHS、CHI、F3H、DFR、ANS、LDOX、UF3GT，调控基因PAP1、PAP2的表达量，关键酶PAL、CHI酶活性较对照相比都有显著提高，提高趋势与花青苷变化趋势相同。由此说明硫化氢通过上调调控花青苷合成的转录因子和关键结构基因的表达，提高关键酶活性来促进花青苷的合成和积累。PAG是硫化氢的合成抑制剂，PAG与NaHS共处理后萝卜芽苗中花青苷含量显著下降但仍高于对照，表明外源硫化氢可能通过介导内源硫化氢含量影响花青苷合成。29788
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毕业论文关键词：花青苷；萝卜芽苗；硫化氢；基因表达
The effects of hydrogen sulfide on anthocyanin biosynthesis in hypocotyls of radish sprouts Student majoring in Horticulture
Abstract: In this study, radish sprouts varieties ‘YangHua’ sensitive to anthocyanin were tested to investigate the effects of hydrogen sulfide (H2S) on the synthesis of flavonoids. The influences of H2S on anthocyanin synthesis were studied with different concentrations of exogenous H2S donor, NaHS, treatments and the absence of NaHS was as control. The results showed that the content of anthocyanin was significantly improved as the increasing of NaHS concentration within a certain range(0.5 mM～2 mM). The measures of structural gene and transcription factor expressions showed that the key structure genes,PAL, CHS, CHI, F3H, DFR, ANS, LDOX, UF3GT,and regulator genes, PAP1, PAP2 of anthocyanin synthesis pathway were notably promoted as the increasing tendency of anthocyanin change. Meanwhile, the crucial enzyme activity of PAL and CHI were higher with NaHS treatments than control. All the conclusions showed that H2S developed anthocyanin synthesis and accumulation by regulating the transcription factors and key structural genes stimulated the enzyme activity. Anthocyanin content in radish seedlings was decreased significantly but still higher than the control with the co-treatment of NaHS and PAG, the inhibitor of H2S synthesis,which showed that exogenous H2S by mediating the endogenous H2S content to regulate anthocyanin synthesis.
Key words: anthocyanin; radish sprouts; H2S; gene expression
目 录
摘要 1
关键词 1
Abstract. 1
Key words 1
引言 1
1 材料与方法 3
1.1 试验时间与试验材料 3
1.2 催芽、播种与培养方法 3
1.3 硫氢化钠浓度的选择与确定 4
1.4 花青苷含量测定方法 4
1.5 苯丙氨酸转氨酶（PAL）和查尔酮异构酶（CHI）活力的测定方法 4
1.5.1 PAL酶活检测方法 4
1.5.2 CHI酶活检测方法 4
1.6 CHS酶活检测方法 4
1.7 基因表达量检测方法 5
1.8 抑制剂浓度的选择与确定 5
1.9 数据处理及作图方法 5
2 结果与分析 5
2.1 外源硫氢化钠对萝卜芽苗下胚轴中花青苷含量的影响 5
2.2 外源硫氢化钠处理对花青苷合成途径中关键酶活力的影响 6