Xu Kedong, Chang Yunxia, Liu Kun, Wang Feige, Liu Zhongyuan, Zhang Ting, Li Tong, Zhang Yi, Zhang Fuli, Zhang Ju, Wang Yan, Niu Wei, Jia Shuzhao, Xie Hengchang, Tan Guangxuan, Li Chengwei
Key Laboratory of Plant Genetics and Molecular Breeding, Zhoukou Normal University, Zhoukou, People's Republic of China.
College of Life Science and Agronomy, Zhoukou Normal University, Zhoukou, People's Republic of China.
PLoS One. 2014 Jun 4;9(6):e98672. doi: 10.1371/journal.pone.0098672. eCollection 2014.
A new protocol was established for the regeneration of Solanum nigrum by frog egg-like bodies (FELBs), which are novel somatic embryogenesis (SE) structures induced from the root, stem, and leaf explants. The root, stem, and leaf explants (93.33%, 85.10%, and 100.00%, respectively) were induced to form special embryonic calli on Murashige and Skoog (MS) medium containing 1.0 mg/L 2,4-dichlorophenoxyacetic acid, under dark condition. Further, special embryonic calli from the root, stem, and leaf explants (86.97%, 83.30%, and 99.47%, respectively) were developed into FELBs. Plantlets of FELBs from the three explants were induced in vitro on MS medium supplemented with 5.0 mg/L 6-benzylaminopurine and 0.1 mg/L gibberellic acid, and 100.00% plantlet induction rates were noted. However, plantlet induction in vivo on MS medium supplemented with 20 mg/L thidiazuron showed rates of 38.63%, 15.63%, and 61.30% for the root, stem, and leaf explants, respectively, which were lower than those of the in vitro culture. Morphological and histological analyses of FELBs at different development stages revealed that they are a novel type of SE structure that developed from the mesophyll (leaf) or cortex (stem and root) cells of S. nigrum.
建立了一种通过类蛙卵体（FELBs）再生龙葵的新方案，类蛙卵体是从根、茎和叶外植体诱导产生的新型体细胞胚胎发生（SE）结构。在含有1.0 mg/L 2,4-二氯苯氧乙酸的Murashige和Skoog（MS）培养基上，在黑暗条件下，根、茎和叶外植体（分别为93.33%、85.10%和100.00%）被诱导形成特殊的胚性愈伤组织。此外，来自根、茎和叶外植体的特殊胚性愈伤组织（分别为86.97%、83.30%和99.47%）发育成类蛙卵体。在添加5.0 mg/L 6-苄基腺嘌呤和0.1 mg/L赤霉素的MS培养基上，体外诱导来自三种外植体的类蛙卵体形成植株，植株诱导率为100.00%。然而，在添加20 mg/L噻苯隆的MS培养基上体内诱导植株时，根、茎和叶外植体的诱导率分别为38.63%、15.63%和61.30%，低于体外培养的诱导率。对不同发育阶段的类蛙卵体进行形态学和组织学分析表明，它们是一种新型的体细胞胚胎发生结构，由龙葵的叶肉（叶）或皮层（茎和根）细胞发育而来。
