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仙客来体细胞胚和合子胚的蛋白质组学分析揭示了种子及萌发生理学的新见解。

Proteomic analyses of somatic and zygotic embryos of Cyclamen persicum Mill. reveal new insights into seed and germination physiology.

作者信息

Winkelmann Traud, Heintz Dimitri, Van Dorsselaer Alain, Serek Margrethe, Braun Hans-Peter

机构信息

Institute of Floriculture and Woody Plant Science, Faculty of Natural Sciences, University of Hannover, Herrenhaeuser Strasse 2, 30419 Hannover, Germany.

出版信息

Planta. 2006 Aug;224(3):508-19. doi: 10.1007/s00425-006-0238-8. Epub 2006 Feb 22.

Abstract

In the horticulturally important ornamental species Cyclamen persicum Mill., somatic embryogenesis is an efficient vegetative propagation method and the development of artificial seeds is an ultimate aim. This study aims at a systematic comparison of the proteomes of zygotic embryos, somatic embryos grown in liquid medium containing 30 or 60 g l(-1) sucrose, germinating embryos of both types and endosperm in order to obtain novel insights into seed and germination physiology. Using high resolution two-dimensional isoelectric focussing/sodium dodecylsulfate polyacrylamide gel electrophoresis (2D IEF/SDS PAGE), 74% of the proteins expressed in zygotic embryos were found in similar abundance in somatic embryos grown in 60 g l(-1) sucrose. Somatic embryos grown in 30 g l(-1) sucrose accumulated fewer protein species than those grown in 60 g l(-1). Selected proteins were identified following mass spectrometry (nano-LC-MS/MS). Four enzymes involved in glycolysis (UDP-glucose pyrophosphorylase, fructose bisphosphate aldolase, triosephosphate isomerase and glyceraldehyde-3-phosphate dehydrogenase GAPDH) were specifically induced in somatic embryos. 11S globulin proteins identified by MS were present in high levels in somatic embryos, zygotic embryos and endosperm, whereas 7S globulins were detected mainly in endosperm and zygotic embryos. These are the first storage proteins identified in C. persicum. Xyloglucans are known to be another group of seed storage compounds in C. persicum. Interestingly, xyloglucan endotransglycosylases were found to be highly expressed in endosperm tissue. We discuss the physiological implications of these observations.

摘要

在园艺学上重要的观赏植物仙客来(Cyclamen persicum Mill.)中,体细胞胚胎发生是一种有效的营养繁殖方法,而人工种子的开发是最终目标。本研究旨在系统比较合子胚、在含有30或60 g l⁻¹蔗糖的液体培养基中生长的体细胞胚、两种类型的萌发胚以及胚乳的蛋白质组,以便获得关于种子和萌发生理学的新见解。使用高分辨率二维等电聚焦/十二烷基硫酸钠聚丙烯酰胺凝胶电泳(2D IEF/SDS PAGE),发现在合子胚中表达的74%的蛋白质在60 g l⁻¹蔗糖中生长的体细胞胚中丰度相似。在30 g l⁻¹蔗糖中生长的体细胞胚积累的蛋白质种类比在60 g l⁻¹蔗糖中生长的体细胞胚少。通过质谱(nano-LC-MS/MS)鉴定了选定的蛋白质。参与糖酵解的四种酶(UDP-葡萄糖焦磷酸化酶、果糖二磷酸醛缩酶、磷酸丙糖异构酶和甘油醛-3-磷酸脱氢酶GAPDH)在体细胞胚中被特异性诱导。通过质谱鉴定的11S球蛋白在体细胞胚、合子胚和胚乳中含量很高,而7S球蛋白主要在胚乳和合子胚中检测到。这些是在仙客来中鉴定出的首批贮藏蛋白。木葡聚糖是仙客来中另一类种子贮藏化合物。有趣的是,发现木葡聚糖内转糖基酶在胚乳组织中高度表达。我们讨论了这些观察结果的生理学意义。

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