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为分析转rolABC基因枳橙GA20ox1基因与rol基因表达的互作关系,进一步阐释其矮化性状形成的分子机制。以转rol基因枳橙实生苗为试验材料,研究其对赤霉素的敏感反应,用荧光定量RT-PCR分析GA20ox1基因和rol基因的表达,并检测幼芽中过氧化物酶活性和植物内源激素含量的变化。结果表明转rol基因枳橙既不属于GA缺陷型,也不属于GA不敏感型,喷施GA3能促进其茎伸长生长,但恢复不到野生型水平,幼芽中IAA(P<0.01)、GA1和GA4(P<0.05)显著降低,POD酶活性显著提高(P<0.01)。转rol基因枳橙幼芽中GA20ox1基因mRNA水平相比对照显著下调(P<0.01)。B、D系与野生型嫩茎中无明显差异,B、D系老叶中明显降低,E系中嫩茎和老叶中均明显增加。B、D和E系嫩叶中GA20ox1基因转录表达均较野生型高。在幼芽、嫩茎中,rolC基因与GA20ox1表达负相关。rol基因通过在幼芽中的高表达下调了GA20ox1基因转录表达,进而抑制了活性GAs在幼芽中的合成,顶端分生组织较低量的活性GAs限制植物茎伸长,在转rolABC基因枳橙矮化性状建成中发挥重要作用。
In order to analyze the interaction between rolABC gene GA20ox1 gene and rol gene expression, we further elucidate the molecular mechanism of dwarfing trait formation. The rol gene transgenic orange citrange was used as the experimental material to study its sensitivity to gibberellin. The expression of GA20ox1 gene and rol gene was analyzed by fluorescence quantitative RT-PCR, and the peroxidase activity Changes in source hormones. The results showed that the rol gene rolong orange was neither GA-deficient nor GA insensitive. Spraying GA3 could promote its stem elongation growth, but it could not recover to the wild-type level. IAA (P <0.01) , GA1 and GA4 (P <0.05), POD activity was significantly increased (P <0.01). The mRNA level of GA20ox1 gene in transgenic rol gene was significantly down-regulated compared with the control (P <0.01). There was no significant difference between B and D lines and wild-type tender stems. The old leaves of B and D lines were significantly reduced, while the tender stem and old leaves of E lines were significantly increased. The transcripts of GA20ox1 gene in young leaves of B, D and E lines were higher than that of wild type. In shoots and young stems, rolC gene was negatively correlated with GA20ox1 expression. rol gene down-regulates the transcriptional expression of GA20ox1 gene through the high expression in shoots, thereby inhibiting the synthesis of active GAs in young shoots. The lower amount of active GAs in the top meristem restricts plant stem elongation. Orange dwarf traits to play an important role in the construction.