Kim T D, Lee B S, Kim T S, Choi Y E
Division of Forest Resources, College of Forest and Environmental Sciences, Kangwon National University, Chunchon 200-701, Korea.
Ann Bot. 2007 Aug;100(2):177-83. doi: 10.1093/aob/mcm094. Epub 2007 Jun 11.
In Tilia amurensis, two types of trichomes (hairy and glandular) develop from epidermal surfaces of cotyledons and hypocotyls of zygotic embryos soon after germination. Here, it is demonstrated that glandular trichome initials develop directly into somatic embryos when treated in vitro with 2,4-dichlorophenoxyacetic acid (2,4-D).
Zygotic embryos of Tilia amurensis were cultured on Murashige and Skoog medium with 3 % sucrose and various concentrations (0, 2.2, 4.4 and 8.8 microm) of 2,4-D. Morphological development of trichomes and somatic embryos was analysed by scanning electron microscope and light microscope after histological sectioning.
In zygotic embryos cultured on medium with 4.4 microM 2,4-D, formation of hairy trichomes was completely suppressed but formation of glandular trichome initials increased. That some filamentous trichome initials developed directly into somatic embryos was confirmed by histological and scanning electron microscope observation. When explants with different stages of trichome initials (two-, four- and eight-celled filamentous and fully mature trichomes) were temporally pre-treated with 4.4 microM 2,4-D for 24 h and transferred into hormone-free medium, two-celled and four-celled filamentous trichome initials were the effective stage of trichomes for somatic embryo induction.
It is suggested that early developing filamentous trichome initials have developmental plasticity and that with 2,4-D treatment these trichome initials develop directly into somatic embryos.
在紫椴中,两种类型的毛状体(毛状和腺状)在合子胚萌发后不久从子叶和下胚轴的表皮表面发育而来。在此,研究表明,用2,4 - 二氯苯氧乙酸(2,4 - D)体外处理时,腺状毛状体原始细胞可直接发育成体细胞胚。
将紫椴合子胚培养在含有3%蔗糖和不同浓度(0、2.2、4.4和8.8微摩尔)2,4 - D的Murashige和Skoog培养基上。在组织切片后,通过扫描电子显微镜和光学显微镜分析毛状体和体细胞胚的形态发育。
在含有4.4微摩尔2,4 - D的培养基上培养的合子胚中,毛状毛状体的形成被完全抑制,但腺状毛状体原始细胞的形成增加。组织学和扫描电子显微镜观察证实,一些丝状毛状体原始细胞直接发育成体细胞胚。当用4.4微摩尔2,4 - D对处于不同毛状体原始细胞阶段(二细胞、四细胞和八细胞丝状以及完全成熟的毛状体)的外植体进行24小时的临时预处理,然后转移到无激素培养基中时,二细胞和四细胞丝状毛状体原始细胞是毛状体诱导体细胞胚的有效阶段。
表明早期发育的丝状毛状体原始细胞具有发育可塑性,并且通过2,4 - D处理,这些毛状体原始细胞可直接发育成体细胞胚。