摘    要 番茄是全球范围内广泛种植的经济作物，也是研究肉质果实发育和成熟的模式作物。近年来，关于转录因子介导的调控网络在番茄果实成熟中的作用，已经取得突破性的进展，其中LeSPL-CNR便是发挥至关重要作用的转录因子。LeSPL-CNR属于SBP-box转录因子家族，LeSPL-CNR启动子区高度甲基化导致LeSPL-CNR转录表达受阻，从而导致无色不成熟突变体Cnr的表型。）。本研究通过实时荧光定量PCR技术分析了野生型AC和突变体Cnr在根、茎、叶、花、果实等不同组织中LeSPL-CNR表达模式发现，不论是在野生型AC还是突变体Cnr中，LeSPL-CNR在果实和花中的表达量远远高于根、茎、叶，这表明相较于番茄的营养生长相比，转录因子LeSPL-CNR主要在果实发育过程中发挥作用。不仅如此，进一步比较LeSPL-CNR在野生型AC和突变体Cnr果实发育不同时期的基因表达模式发现，在AC果实发育和成熟过程中LeSPL-CNR基因表达呈现的趋势为：先升高，至破色期达到顶峰，然后又迅速下降；而在突变体Cnr中，LeSPL-CNR基因表达呈现逐渐降低的趋势，并且基因表达丰度始终保持在非常低的水平，较AC中的表达有差异显著。以上结果进一步证实果实内低丰度的LeSPL-CNR表达水平是导致突变体Cnr不成熟表型的主要原因，将有助于阐释LeSPL-CNR在调控番茄果实发育和成熟的生物学功能。90261

毕业论文关键词：番茄；实时荧光定量PCR；LeSPL-CNR；突变体Cnr 

Abstract   Tomato is an economic crop cultivated globally wide, and has long been the ideal model of fleshy fruit development and ripening。 In recent years, the role of transcription factor-mediated regulatory networks during the fruit ripening in tomato has made breakthrough progress, in which LeSPL-CNR is a transcription factor that plays a vital role。 LeSPL-CNR belongs to the family of SBP-box transcription factors, the highly-methylated region in the promotor of LeSPL-CNR inhibited the expression of LeSPL-CNR gene expression, which resulted in the phenotype of colorless-non ripening mutant Cnr。 Therefore, the expression pattern of LeSPL-CNR in wild type AC and mutant Cnr was analyzed by qRT-PCR, respectively。 on the one hand, via comparing the expression pattern of LeSPL-CNR in different tissues, it is found that the expression level of LeSPL-CNR was much higher in the flowers and fruits than that in the root, stem, and leaves, not only in AC but also in Cnr, which suggested that the role of LeSPL-CNR during fruit development and ripening was more important than that during vegetative growth。 On the other hand, via comparing the differential and developmental expression of LeSPL-CNR in AC and Cnr mutant fruits, the relative expression of LeSPL-CNR trended to increase gradually during AC fruits development, reached a peak at the breaker stage, and followed by a rapid decline when fruits started to ripen, whereas in Cnr mutant fruits, expression levels of LeSPL-CNR reduced substantially and remained a very low level throughout all the developmental stages, which had a significant difference than that in AC。 These results conformed again that the lowly expression level of LeSPL-CNR led to the phenotype of Cnr mutant, which will help to elucidate the biological role of LeSPL-CNR during the fruit development and ripening in tomato。

  Key words: tomato；Real-time quantitative PCR；LeSPL-CNR；mutant Cnr

目    录

摘要 3

Abstract 4

目录 5

1。 引言 6

1。1 定量PCR技术 6

1。1。1 定量PCR技术的由来