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休眠和非休眠野燕麦颖果胚胎中差异表达基因的cDNA克隆的表征

Characterization of cDNA clones for differentially expressed genes in embryos of dormant and nondormant Avena fatua L. caryopses.

作者信息

Johnson R R, Cranston H J, Chaverra M E, Dyer W E

机构信息

Department of Plant, Soil and Environmental Sciences, Montana State University, Bozeman 59717-0312, USA.

出版信息

Plant Mol Biol. 1995 Apr;28(1):113-22. doi: 10.1007/BF00042043.

DOI:10.1007/BF00042043
PMID:7787176
Abstract

The molecular regulation of seed dormancy was investigated using differential display to visualize and isolate cDNAs representing differentially expressed genes during early imbibition of dormant and nondormant Avena fatua L. embryos. Of about 3000 cDNA bands examined, 5 cDNAs hybridized with mRNAs exhibiting dormancy-associated expression patterns during the first 48 h of inhibition, while many more nondormancy-associated cDNAs were observed. Dormancy-associated clone AFD1 hybridized with a 1.5 kb mRNA barely detectable in dry dormant and nondormant embryos that became more abundant in dormant embryos after 24 h of imbibition. Clone AFD2 hybridized with two mRNAs, a 1.3 kb message constitutively expressed in dormant and nondormant embryos and a 0.9 kb message present at higher levels in dormant embryos after 3 h of imbibition. Nondormancy-associated clones AFN1, AFN2 and AFN3 hybridized with 1.5 kb, 1.7 kb and 1.1 kb mRNAs, respectively, that were more abundant in nondormant embryos during imbibition. Expression patterns of some mRNAs in dormant embryos induced to germinate by GA3 treatment were different than water controls, but were not identical to those observed in nondormant embryos. DNA sequence analysis revealed 76% sequence identity between clone AFN3 and a Citrus sinensis glutathione peroxidase-like cDNA, while significant sequence similarities with known genes were not found for other clones. Southern hybridization analyses showed that all clones represent low (1 to 4) copy number genes.

摘要

利用差异显示技术研究种子休眠的分子调控,以可视化并分离代表休眠和非休眠野燕麦胚胎早期吸水过程中差异表达基因的cDNA。在检测的约3000条cDNA条带中,有5条cDNA与在吸水抑制的前48小时内呈现与休眠相关表达模式的mRNA杂交,同时观察到更多与非休眠相关的cDNA。与休眠相关的克隆AFD1与一个1.5 kb的mRNA杂交,该mRNA在干燥的休眠和非休眠胚胎中几乎检测不到,在吸水24小时后在休眠胚胎中变得更加丰富。克隆AFD2与两条mRNA杂交,一条1.3 kb的信息在休眠和非休眠胚胎中组成性表达,另一条0.9 kb的信息在吸水3小时后在休眠胚胎中含量更高。与非休眠相关的克隆AFN1、AFN2和AFN3分别与1.5 kb、1.7 kb和1.1 kb的mRNA杂交,这些mRNA在吸水过程中在非休眠胚胎中更为丰富。经GA3处理诱导发芽的休眠胚胎中一些mRNA的表达模式与水对照不同,但与非休眠胚胎中观察到的模式也不相同。DNA序列分析显示克隆AFN3与柑橘谷胱甘肽过氧化物酶样cDNA之间有76%的序列同一性,而其他克隆未发现与已知基因有显著的序列相似性。Southern杂交分析表明,所有克隆均代表低(1至4)拷贝数基因。

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