DOI: 10.3724/SP.J.1006.2017.01417

Acta Agronomica Sinica (作物学报) 2017/43:10 PP.1417-1425

Cloning of β-amylase Gene (CsBAM3) and Its Expression Model Response to Cold Stress in Tea Plant

A major enzyme of starch breakdown in plant leaf, β-amylase (BAM), plays important roles in response to abiotic stresses. In tea plant, a differentially expressed gene involved in starch metabolism was identified from previous RNA-Seq analysis of cold acclimation. Its full-length cDNA was cloned and sequenced, showing that it is a homolog of BAM3 in Arabidopsis (named CsBAM3). CsBAM3 contains 548 amino acid residues and is grouped into subfamily Ⅱ together with BAM1 and BAM3 from Arabidopsis. According to protein sequence and 3D structure analyses, CsBAM3 was considered as a chloroplast-targeted protein with β-amylase activity. Promoter cloning and sequence analysis showed a couple of responsive elements related to circadian, light, cold, and phytohormone, indicating its complex regulating mechanism. CsBAM3 showed the expression higher in leaf, relative lower in stem and flower and extremely low in root. The expression of CsBAM3 in mature leaf was significantly up-regulated during the early time courses of winter cold acclimation and maintained at relative high level during the following period. With the treatments of 4℃ and 0℃, the transcript levels of CsBAM3 in both mature leaves and young shoots were dramatically increased, interestingly, much higher level was detected in young shoots. The cold environment similar to spring cold spell was created using climate chamber. And the expression changes of CsBAM3 in shoots were determined then after the tea plants were moved into climatic chamber at different sprouting phrases. CsBAM3 was quickly up-regulated by cold once the first leaf unfolded. Our study illuminated that CsBAM3 is a key β-amylase coding gene involved in starch metabolism of tea plant, which can be stimulated in both mature leaves and young shoots by cold stress.

Key words:β-amylase,Tea plant,Gene clone,Cold stress,Expression regulation

ReleaseDate:2017-12-07 14:03:20

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