Key Laboratory of Resource Biology and Biotechnology in Western China (Ministry of Education), Shaanxi Provincial Key Laboratory of Biotechnology, College of Life Sciences, Northwest University, Xi'an, Shaanxi 710069, People's Republic of China.
Key Laboratory of Molecular Engineering of Polymers of Ministry of Education, Department of Macromolecular Science and Laboratory of Advanced Materials, Fudan University, Shanghai 200433, People's Republic of China.
J Plant Physiol. 2019 Sep;240:152991. doi: 10.1016/j.jplph.2019.152991. Epub 2019 Jun 8.
The coding sequence of NtabSPL6-1 was cloned by high-fidelity PCR with specific primers and was used in construction of a binary vector for overexpression. Wild-type Col-0 Arabidopsis plants and Qinyan95 tobacco leaves were transformed using floral dip and leaf disc methods, respectively. Phenotypic observation showed that constitutive expression of NtabSPL6-1 in Arabidopsis could promote the development of trichomes on leaf epidermis and influence the growth pattern of cauline leaves. In tobacco, ectopic expression of NtabSPL6-1 led to dwarfism of the plants and alteration of the leaf structure, accompanied by changes of the glandular trichomes in development. At the same time, the self-regulation capability of NtabSPL6-1 was determined by yeast two-hybrid system. The results indicated that SBP-C terminal domain and C terminal domain of NtabSPL6-1 possessed strong transcriptional activation ability; the intact protein, N terminal domain, and the first peptide fragment in N terminal domain possessed weak transcriptional activation ability; and the second and the third peptide fragments in N terminal domain had no transcriptional activation ability, suggesting the N terminal domain of NtabSPL6-1 could block the activity of the C terminal domain. NtabSPL6-1 may affect the resistance of plants to biotic stress factors indirectly by regulation of the trichome growth.
NtabSPL6-1 的编码序列通过特异性引物的高保真 PCR 进行克隆，并用于构建用于过表达的二元载体。使用花浸和叶盘法分别转化野生型 Col-0 拟南芥植物和 Qinyan95 烟草叶片。表型观察表明，NtabSPL6-1 在拟南芥中的组成型表达可以促进叶表皮毛的发育，并影响茎生叶的生长模式。在烟草中，NtabSPL6-1 的异位表达导致植物矮化和叶片结构改变，同时伴随着发育中的腺毛的变化。同时，通过酵母双杂交系统确定了 NtabSPL6-1 的自我调节能力。结果表明，NtabSPL6-1 的 SBP-C 端结构域和 C 端结构域具有很强的转录激活能力；完整蛋白、N 端结构域和 N 端结构域的第一个肽片段具有较弱的转录激活能力；而 N 端结构域的第二个和第三个肽片段没有转录激活能力，表明 NtabSPL6-1 的 N 端结构域可以阻断 C 端结构域的活性。NtabSPL6-1 可能通过调节毛发生长间接影响植物对生物胁迫因子的抗性。
