APETALA1大豆同源异型基因GmAPETALA1的克隆及在光周期诱导条件下的表达

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	APETALA1大豆同源异型基因GmAPETALA1的克隆及在光周期诱导条件下的表达
Alternative Title	Cloning GmAP1and analyzing its Expression patterns during floral reversion induced by changes of photoperiod in soybean development
	高小伟
Thesis Advisor	王根轩
	2008-12-11
Degree Grantor	兰州大学
Place of Conferral	兰州
Degree Name	博士
Keyword	光周期自贡冬豆长日照短日照顶端分生组织开花逆转兼并引物
Abstract	本文试验的出发点是一个传统的生态学问题。本文在立足于生态学这样一个基本生态问题—植物如何适应光周期现象，而后延伸到了光周期适应物种是如何适应这种光周期现象而进行程序性发育？光照和温度是自然界中两大最基本的生态因子。从根本上讲，物种（尤其是植物）的进化就是适应光温条件的一个过程。中国地域辽阔，纬度跨越很大，因此在农业生产和品种选育过程中，各个地方逐渐形成了自己的栽培品种，而这些栽培品种无非就是单一适应地方光温条件。本文所用材料为自贡冬豆（Glycine max[L.] Merr.）,大豆中晚熟品种，短日植物，产于四川自贡地区，对光周期反应敏感。在室内模拟纬度变化条件下的光周期变化，在不同光周期条件下比较观察自贡冬豆的生长形态，我们发现自贡冬豆存在三种典型的形态：持续短日（12h/d）条件下的正常花；持续长日（16h/d）完全营养生长；短日/长日条件下花序呈现（自上而下）营养芽、逆转花和正常花混合分布的现象。这些现象说明了四个方面的问题：第一适宜的光周期条件是自贡冬豆成功地由营养生长向生殖生长转变的必须生态因子；第二自贡冬豆由营养生长向生殖生长转变所需光周期条件是连续的；第三维持自贡冬豆开花的信号具有数量特征；第四这些特征同模式植物拟南芥生长发育转变所必须的AP1（APETALA1）花分生组织特征基因行使功能赋予的特征相似。 APETALA1 基因是MADS-box 基因家族中的成员，它们具有高度保守的结构域在植物的发育过程中功能又很高的保守性。根据这些特征结合双子叶植物碱基使用的保守性，我们设计了兼并引物并成功调取了APETALA1 在大豆中的同源类似基因（homolog）GmAPETALA1。在碱基水平和蛋白水平上，GmAPETALA1 同APETALA1 的相似性分别为45%和56.08%。 RT-PCR 实验证明在长日条件，GmAPETALA1 在顶端分生组织（shoot apical meristem）表达处于很低的水平，并且表达水平不会随着长日条件的改变而改变；在短日条件下GmAPETALA1 在顶端分生组织中的表达随着短日条件的延长而升高并且在花器官中的表达处于很高的水平；不论在长短日条件下，在其它组织中都没有表达。为了分析APETALA1 在大豆中的表达，我们比较分析了几种不同的方法制作核酸探针的效率并发现利用PCR 的方法制作ssDNA 探针与利用转录的方法制作的 RNA 探针以及PCR 方法制作的dsDNA 探针相比较可以获得更高产量。关键词：光周期；自贡冬豆；短日照（12h/d）；长日照（16h/d）; 兼并引物； GmAPETALA1; RT-PCR; 顶端分生组织；探针
Other Abstract	The experiments included in this paper are derived from a general ecological question. Based on photoperiodism, a question was asked that how plant species adapt to photoperiod and develop stereotypcially and properly? Light and temperature are two basally ecological factors. Intrincically, species (especially plant) evolution is a progress to adapt to preferential light and temperature. In china, covered widly, many cultival species are those narrow light- and temperature-adaptation species. The experimental plant is Zigongdongdou (Glycine max[L.] Merr.), which is a cultival species, characterized with late maturation, short day—induced reproduction and high sensitivity to photoperiod. Previous experiments showed that inflorescences of Zigongdongdou took on three kinds of typical phenotypes, including normal flowers under continuous short-day light(12h/d); the whole vegetative state of shoot apical meristem under continuous long-day light(16h/d) and fused phenotypes with apical-to-basal vegetative leaves、reversed flowers and normal flowers when plants were transfer from short day(enough time to induce floral initiation) to long day condition. These results suggest that correct photoperiod is necessary for successful transition of plant development stage from vegetative stage to reproductive stage; the needed photoperiod is continuous; signals maintenning floral transition is quantitative and these phenotypes caused by variable photoperiod is Arabidopsis. APETALA1 is a member of MADS-box family with conserved domains for plant development. In combination with properties of dicotyledons preferring to bases, degenerate primers were designed. Using the pair of primers, GmAPETALA1, homolog of APETALA1 in soybean was cloned. The similarity of the cDNA and protein sequence to those of APETALA1 was 45% and 56.08% respectively. RT-PCR was conducted and showed that at long-day condition, GmAPETALA1 expression level was very low and kept low level in shoot apical meristem even increasing long-day treatment; at short-day condition, the expression level increased with more short-day treatment and have a higher level in floral organs; its expression did not expand to other tissues no matter at what conditions. For more expriments, we synthesized DIG-labelled probes using different protocols and checked their synthesis efficience. The results showed that PCR-based synthesis of ssDNA had a higher efficience than transcription-based synthesis RNA probes and PCR-base...
URL	查看原文
Language	中文
Document Type	学位论文
Identifier	https://ir.lzu.edu.cn/handle/262010/222075
Collection	生命科学学院
Recommended Citation GB/T 7714	高小伟. APETALA1大豆同源异型基因GmAPETALA1的克隆及在光周期诱导条件下的表达[D]. 兰州. 兰州大学,2008.

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