Genome-wide identification of BAM genes in grapevine (Vitis vinifera L.) and ectopic expression of VvBAM1 modulating soluble sugar levels to improve low-temperature tolerance in tomato.
Genome-wide identification of BAM genes in grapevine (Vitis vinifera L.) and ectopic expression of VvBAM1 modulating soluble sugar levels to improve low-temperature tolerance in tomato.
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DOI：
10.1186/s12870-021-02916-8
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发表时间：
2021-03-26
影响因子：
5.3
通讯作者：
Mao J
中科院分区：
文献类型：
--
作者：
Liang G;He H;Nai G;Feng L;Li Y;Zhou Q;Ma Z;Yue Y;Chen B;Mao J
低温是影响葡萄生长发育的主要限制因子之一。增加可溶性糖和清除活性氧(ROS)在葡萄抗寒中起关键作用。然而，β-淀粉酶(BAM)参与调节糖水平和抗氧化酶活性以响应冷胁迫的机制尚不清楚。在本研究中，共鉴定了6个BAM基因，并将其聚为4个组。多重序列比对和基因结构分析表明，VvBAM6缺少Glu380残基，只含有一个外显子。随着温度的降低，VvBAM1和VvBAM3的转录丰度显著增加。在LT胁迫后，VvBAM1在高表达番茄品系的叶片、叶柄、茎和根中高表达。与野生型番茄相比，转基因植株的总淀粉酶活性和BAM活性分别提高了6.5和6.01倍。在LT胁迫下，转基因植株的葡萄糖和蔗糖含量显著高于WT植株，而前者的淀粉含量比后者低1.5倍。转录组测序数据分析表明，转基因植株中有541个基因表达上调，663个基因表达下调。1个糖转运蛋白基因(SLSTP10)、2个过氧化物酶(POD)相关基因(SLPER7和SLPER5)和1个过氧化氢酶(CAT)相关基因(SlCAT1)在LT胁迫后分别上调8.6、3.6、3.0和2.3倍。我们的结果表明，VvBAM1的过表达促进了对ROS的清除，并通过调节淀粉水解来影响番茄植株的可溶性糖水平，从而提高了番茄的耐冷性。网上版载有补充材料，可在10.1186/s12870-021-02916-8查阅。
Low temperature (LT) is one of the main limiting factors that affect growth and development in grape. Increasing soluble sugar and scavenging reactive oxygen species (ROS) play critical roles in grapevine resistance to cold stress. However, the mechanism of β-amylase (BAM) involved in the regulation of sugar levels and antioxidant enzyme activities in response to cold stress is unclear. In this study, six BAM genes were identified and clustered into four groups. Multiple sequence alignment and gene structure analysis showed that VvBAM6 lacked the Glu380 residue and contained only an exon. The transcript abundance of VvBAM1 and VvBAM3 significantly increased as temperature decreased. After LT stress, VvBAM1 was highly expressed in the leaves, petioles, stems, and roots of overexpressing tomato lines. The total amylase and BAM activities increased by 6.5- and 6.01-fold in transgenic plants compared with those in wild-type tomato plants (WT) subjected to LT, respectively. The glucose and sucrose contents in transgenic plants were significantly higher than those in WT plants, whereas the starch contents in the former decreased by 1.5-fold compared with those in the latter under LT stress. The analysis of transcriptome sequencing data revealed that 541 genes were upregulated, and 663 genes were downregulated in transgenic plants. One sugar transporter protein gene (SlSTP10), two peroxidase (POD)-related genes (SlPER7 and SlPER5), and one catalase (CAT)-related gene (SlCAT1) were upregulated by 8.6-, 3.6-, 3.0-, and 2.3-fold in transgenic plants after LT stress, respectively. Our results suggest that VvBAM1 overexpression promotes ROS scavenging and improves cold tolerance ability by modulating starch hydrolysis to affect soluble sugar levels in tomato plants. The online version contains supplementary material available at 10.1186/s12870-021-02916-8.
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