Cloning and Expression Analysis of the ScD53 Gene from Sugarcane
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D53 is a crucial gene involved in tillering regulation in crops via the strigolactone signaling pathway. The objective of our research was to clone the homologous sugarcane (Saccharum L. spp. hybrids) gene of D53 (ScD53) by in silico cloning combined with RT-PCR and rapid amplification of cDNA ends techniques, and the physicochemical properties and structure of ScD53 protein were analyzed using bioinformatic software and tools. The relationships between the expression of ScD53 and some plant hormones, bud germination and tillering occurrence of seedlings were also detected using the real-time fluorescence quantitative PCR (FQ-PCR) technique. The homolog was named ScD53 and possessed a 3918-bp cDNA sequence. Bioinformatic analysis showed that ScD53 has a complete open reading frame of 3363 bp that encodes 1120 amino acid residues. The ScD53 protein possesses the conserved domain of the P-loop NTPase and ClpB_D2-small superfamilies, which may be unstable hydrophilic proteins and are likely to be located in the nucleus. The main secondary and tertiary structures of the ScD53 protein are composed of random coil and alpha helix structures and exhibit high similarity in structure and homology with D53 proteins from sorghum [Sorghum bicolor (L.) Moench] and maize (Zea mays L.). The FQ-PCR results showed that the expression of ScD53 is tissue specific and is differently expressed in the different tissues. The expression of ScD53 is associated with bud germination, the high expression of which inhibits bud germination. Plant hormone treatment experiments indicated that the expressions of ScD53 were induced in bud and leaf tissues by strigolactones, 3-indole acetic acid and kinetin (KT, a cytokinin analog). The high expression of ScD53 following strigolactone and auxin treatment obviously delayed occurrence of tillers and reduced the number of tillers for the seedlings. However, this was not observed during KT treatment, which implied that the negative effects of this gene on seedlings tillering phenotype were eliminated by the positive influences of other unknown pathways. Our results above help elucidate the function of ScD53 and provide important information and direction for the use of this gene in improving sugarcane yield through tiller regulation.
KeywordsSugarcane ScD53 Cloning Expression Tillering
This work was supported by the National Natural Science Foundation of China (31760412) and Candidates of the Young and Middle Aged Academic Leaders of Yunnan Province (2014HB038), and by funding from the Yunnan Provincial Science and Technology Department for High-End Talent Program. We would like to thank LetPub (www.letpub.com) and Dr. David M. Burner for editorial assistance.
ALL carried out the experiment and wrote the manuscript. XJL, CJL, HBL, QYZ, XQL participated in experimental work. XLL conceived and designed the study.
Compliance with Ethical Standards
Conflict of interest
The authors declare no conflict of interest.
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