Plasma Membrane H(+)-ATPase SmPHA4 Negatively Regulates the Biosynthesis of Tanshinones in Salvia miltiorrhiza.
Plasma Membrane H(+)-ATPase SmPHA4 Negatively Regulates the Biosynthesis of Tanshinones in Salvia miltiorrhiza.
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DOI：
10.3390/ijms22073353
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发表时间：
2021-03-25
影响因子：
5.6
通讯作者：
Dong J
中科院分区：
文献类型：
--
作者：
Li X;Zhang B;Ma P;Cao R;Yang X;Dong J
丹参因其丹参酮等活性成分的药理作用而被广泛应用于心脑血管疾病的治疗。质膜H ~+-ATPase在植物的许多生理过程中起着重要作用。然而，对S.丹参（Sm.）在此，鉴定了9种PM H+-ATP酶同工型，并命名为SmPHA 1-SmPHA 9。系统进化树分析表明，在S.丹参PM H+-ATPase家族。此外，跨膜结构富含SmPHA蛋白。此外，SmPHA 4被发现在根和花中高度表达。高效液相色谱法显示，二氢丹参酮（DT），隐丹参酮（CT）和丹参酮I（TI）的积累显着减少，但增加SmPHA 4-RNAi株系，范围从2.54至3.52，3.77至6.33，和0.35至0.74 mg/g，分别，表明SmPHA 4是丹参酮代谢产物的候选调节剂。此外，qRT-PCR证实，丹参酮生物合成相关关键酶的表达在SmPHA 4-RNAi株系中也上调。总之，本研究揭示了质膜H+-ATP酶的功能，并为研究调控次生代谢生物合成的候选基因提供了新的思路。丹参。
Salvia miltiorrhiza Bunge has been widely used in the treatment of cardiovascular and cerebrovascular diseases, due to the pharmacological action of its active components such as the tanshinones. Plasma membrane (PM) H+-ATPase plays key roles in numerous physiological processes in plants. However, little is known about the PM H+-ATPase gene family in S. miltiorrhiza (Sm). Here, nine PM H+-ATPase isoforms were identified and named SmPHA1–SmPHA9. Phylogenetic tree analysis showed that the genetic distance of SmPHAs was relatively far in the S. miltiorrhiza PM H+-ATPase family. Moreover, the transmembrane structures were rich in SmPHA protein. In addition, SmPHA4 was found to be highly expressed in roots and flowers. HPLC revealed that accumulation of dihydrotanshinone (DT), cryptotanshinone (CT), and tanshinone I (TI) was significantly reduced in the SmPHA4-OE lines but was increased in the SmPHA4-RNAi lines, ranging from 2.54 to 3.52, 3.77 to 6.33, and 0.35 to 0.74 mg/g, respectively, suggesting that SmPHA4 is a candidate regulator of tanshinone metabolites. Moreover, qRT-PCR confirmed that the expression of tanshinone biosynthetic-related key enzymes was also upregulated in the SmPHA4-RNAi lines. In summary, this study highlighted PM H+-ATPase function and provided new insights into regulatory candidate genes for modulating secondary metabolism biosynthesis in S. miltiorrhiza.
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