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Cloning and expression of NtTFL1-1 and NtTFL1-2 genes from Narcissus tazetta var. chinensis*(PDF)

Chinese Journal of Applied & Environmental Biology[ISSN:1006-687X/CN:51-1482/Q]

2020 02
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Cloning and expression of NtTFL1-1 and NtTFL1-2 genes from Narcissus tazetta var. chinensis*
ZHANG Lu1 WU Jinghua1** YAO Xinting1 Gefu WANG-PRUSKI 1 2 & ZHANG Zhizhong 1**
1 Joint FAFU-Dalhousie Lab, College of Horticulture, Fujian Agriculture and Forestry University, Fuzhou, 350002, China
2 Faculty of Agriculture, Dalhousie University, Truro, B2N 5E3, Canada
Narcissus tazetta var. chinensis TFL1 gene flower bud differentiation qPCR subcellular localization

Objectives: Narcissus tazetta var. chinensis ( Nt) is one of the ten famous traditional flowers. It blossoms only in winter and is a typical Spring Festival flowers. The fixed florescence limits its application value. The flower bud differentiation of Nt occurs in the dormancy period of the bulb in summer. This period is the key stage of the research of flowering time regulator. However, the study on flowering mechanism and regulation is still in its infancy. TFL1 is an important flowering-related gene in plants. In order to reveal the role of TFL1 gene in flower bud differentiation and flowering of Nt, we cloned the TFL1 gene from Nt and analyzed its related bioinformatics. TFL1 protein subcellular localization and TFL1 gene expression level were analyzed. The results will be helpful to provide references for florescence regulation and breeding of different florescence Nt varieties. Methods: T he TFL1 gene of Nt were cloned by Reverse Transcription-Polymerase Chain Reaction (RT-PCR) using Nt cv. ‘ Jinzhanyintai’ as material. Two clones were obtained in this experiment and named NtTFL1-1 and NtTFL1-2 . The obtained gene sequence was analyzed by bioinformatics, and the subcellular localization of the gene encoding protein was carried out. The differential expression of the gene in different stages of flower bud differentiation and different organ types of Nt was determined by qPCR. Results: Bioinformatics analysis showed that the ORF of NtTFL1-1 was 498 bp, encoding 166 amino acids, and the ORF of NtTFL1-2 was 522 bp, encoding 174 amino acids, all of which had conserved domains of the TFL1 family. NtTFL1-1 and NtTFL1-2 were localized in the cytoplasm. The results of qRT-PCR analysis showed that the expression of NtTFL1-1 and NtTFL1-2 genes was lower in the early stage of flower bud differentiation and inflorescence primordium formation and higher in flower primordium formation. The expression of NtTFL1-1 and NtTFL1-2 in roots was significantly higher than that in stems, leaves and mature flowers. Conclusions: The results confirmed that NtTFL1-1 and NtTFL1-2 expressed differently in the flower bud differentiation stage of Nt, which were closely related to the flower bud differentiation of Nt. The experimental results could provide theoretical reference for flower bud regulation of Nt.




Last Update: 2019-09-04