Identification and functional characterization of a new flavonoid synthase gene MdFLS1 from apple.
Identification and functional characterization of a new flavonoid synthase gene MdFLS1 from apple.
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苹果新黄酮合酶基因 MdFLS1 的鉴定和功能表征
DOI：
10.1007/s00425-021-03615-2
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发表时间：
2021
期刊：
影响因子：
4.3
通讯作者：
Ji Lusha
中科院分区：
文献类型：
--
作者：
Li Pan;Lei Kang;Liu Lin;Zhang Guizhi;Ge Hongjuan;Zheng Chengchao;Shu Huairui;Zhang Shizhong;Ji Lusha
主要结论苹果类黄酮合成酶基因MdFLS 1在苹果“粉红穗”紫红色枝条中积累，可能在花青苷合成中起重要作用。本研究从青岛市农业科学院苗圃中筛选出20个苹果新品系，其中绿色10个，红色10个。代谢产物分析表明，‘粉红尖顶’紫穗槐紫红色枝条中积累了大量的花色苷。实时荧光定量聚合酶链反应显示，黄酮醇合成酶基因MdFLS 1在M.粉红色的尖顶比其他品种。在MdFLS 1的活性结合位点的前三个碱基处插入一个碱基A，证明苹果叶片和茎的紫红色在M.“粉红色螺旋”可能是MdFLS 1蛋白失活所致。体外酶促反应结果表明，MdFLS 1蛋白丧失了活性。获得高表达野生型菌株（OE 1和OE 2）和A碱基插入高表达菌株（A-OE 1和A-OE 2）。与野生型相比，过量表达株系表现出更浅的组织颜色和更少的花青素积累。而A-OE 1和A-OE 2在着色方面没有差异。因此，我们推测MdFLS 1基因在M.“粉红色尖顶”不能结合类黄酮，触发类黄酮代谢途径的另一个分支中花青素的合成，导致苹果叶片和茎呈紫红色。这些结果表明，MdLS 1基因可能是未来苹果高黄酮育种的一个潜在遗传目标。
Main conclusionThe flavonoid synthase gene MdFLS1 from apple, which possibly plays an important role in anthocyanin synthesis, accumulates in the purple-red branches of Malus ‘Pink spire’.AbstractFlavonoid metabolism serves an important function in plant growth and development. In this study, we selected 20 varieties of apple lines, 10 green and ten red branches, from the plant nursery of Qingdao Agriculture Academy. Metabolite analysis revealed that large amounts of anthocyanins accumulated in the purple-red branches ofM. ‘Pink spire’. Real-time polymerase chain reaction showed that the expression of the flavonol synthase geneMdFLS1was over 1500-fold higher inM.‘Pink spire’ than in the other varieties. A single base A was inserted at the first three bases of the active binding site ofMdFLS1to prove that the purple-red colour of apple leaves and stems inM.‘Pink spire’ may be caused by the inactivation of MdFLS1 protein. The results of in vitro enzymatic reaction revealed that the MdFLS1 protein lost its activity.MdFLS1was expressed inArabidopsis thalianato explore further its functions. High-expression wild-type strains (OE1andOE2) and high-expression strains of A-base insertion (A-OE1andA-OE2) were obtained. Compared with the wild-type strains, the overexpression lines showed lighter tissue colour and less accumulation of anthocyanins. However,A-OE1andA-OE2showed no difference in colouration. In conclusion, we speculated that theMdFLS1gene inM. ‘Pink spire’ cannot bind flavonoids, triggering the synthesis of anthocyanins in another branch of the flavonoid metabolic pathway and resulting in the purple-red colouration of apple leaves and stems. These results suggest thatMdLS1is a potential genetic target for breeding high-flavonoid apples in future cultivar development.