Bi Zhenghong, Li Xiang, Huang Huasun, Hua Yuwei
College of Agronomy, Hainan University, Haikou 570228, China.
Key Laboratory of Rubber Biology of the Ministry of Agriculture, Rubber Research Institute, Chinese Academy of Tropical Agricultural Sciences, Danzhou 571737, China.
Int J Mol Sci. 2016 Mar 2;17(3):247. doi: 10.3390/ijms17030247.
A homolog of MOTHER OF FT AND TFL1 (MFT) was isolated from Hevea brasiliensis and its biological function was investigated. Protein multiple sequence alignment and phylogenetic analysis revealed that HbMFT1 conserved critical amino acid residues to distinguish MFT, FLOWERING LOCUS T (FT) and TERMINAL FLOWER1 (TFL1)-like proteins and showed a closer genetic relationship to the MFT-like group. The accumulation of HbMFT1 was generally detected in various tissues except pericarps, with the highest expression in embryos and relatively higher expression in roots and stems of seedlings, flowering inflorescences, and male and female flowers. HbMFT1 putative promoter analysis showed that tissue-specific, environmental change responsive and hormone-signaling responsive elements were generally present. HbMFT1 was strongly induced under a short-day condition at 28 °C, with the highest expression after the onset of a day. Overexpression of HbMFT1 inhibited seed germination, seedling growth, and flowering in transgenic Arabidopsis. The qRT-PCR further confirmed that APETALA1 (AP1) and FRUITFULL (FUL) were drastically down-regulated in 35S::HbMFT1 plants. A histochemical β-glucuronidase (GUS) assay showed that HbMFT1::GUS activity was mainly detected in stamens and mature seeds coinciding with its original expression and notably induced in rosette leaves and seedlings of transgenic Arabidopsis by exogenous abscisic acid (ABA) due to the presence of ABA cis-elements in HbMFT1 promoter. These results suggested that HbMFT1 was mainly involved in maintenance of seed maturation and stamen development, but negatively controlled germination, growth and development of seedlings and flowering. In addition, the HbMFT1 promoter can be utilized in controlling transgene expression in stamens and seeds of rubber tree or other plant species.
从巴西橡胶树中分离出了FT和TFL1基因的同源基因(MFT),并对其生物学功能进行了研究。蛋白质多序列比对和系统发育分析表明,HbMFT1保留了区分MFT、成花素(FT)和类终端花1(TFL1)蛋白的关键氨基酸残基,并且与类MFT组具有更近的遗传关系。除果皮外,在巴西橡胶树的各种组织中普遍检测到HbMFT1的积累,在胚胎中表达量最高,在幼苗的根和茎、开花的花序以及雄花和雌花中表达量相对较高。对HbMFT1推定启动子的分析表明,通常存在组织特异性、环境变化响应和激素信号响应元件。在28℃短日照条件下,HbMFT1被强烈诱导,在一天开始后表达量最高。在转基因拟南芥中过表达HbMFT1会抑制种子萌发、幼苗生长和开花。定量逆转录聚合酶链反应(qRT-PCR)进一步证实,在35S::HbMFT1植株中,APETALA1(AP1)和FRUITFULL(FUL)显著下调。组织化学β-葡萄糖醛酸酶(GUS)分析表明,HbMFT1::GUS活性主要在雄蕊和成熟种子中检测到,与其原始表达一致,并且由于HbMFT1启动子中存在脱落酸(ABA)顺式元件,在转基因拟南芥的莲座叶和幼苗中被外源ABA显著诱导。这些结果表明,HbMFT1主要参与种子成熟和雄蕊发育的维持,但对种子萌发、幼苗生长和发育以及开花起负调控作用。此外,HbMFT1启动子可用于控制橡胶树或其他植物物种雄蕊和种子中的转基因表达。