The MAP-Kinase HOG1 Controls Cold Adaptation in Rhodosporidium kratochvilovae by Promoting Biosynthesis of Polyunsaturated Fatty Acids and Glycerol
The MAP-Kinase HOG1 Controls Cold Adaptation in Rhodosporidium kratochvilovae by Promoting Biosynthesis of Polyunsaturated Fatty Acids and Glycerol
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DOI：
10.1007/s00284-022-02957-8
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发表时间：
2022-09-01
影响因子：
2.6
通讯作者：
Zhang, Qi
中科院分区：
文献类型：
--
作者：
Chen, Wei;Zhang, Xiaoqing;Zhang, Qi
本研究旨在探讨RKHog1在克氏红孢子虫（Rhodosporidium kratochvilovae）菌株YM25235冷适应中的作用，并阐明其多不饱和脂肪酸（PUFAs）和甘油的生物合成与其冷适应的相关性。对YM25235菌株进行了耐盐性、渗透性和耐冷性分析。反转录实时荧光定量PCR检测YM25235中RKhog1、Delta（12/15）脂肪酸去饱和酶基因（RKD12）、RKMsn4、HisK2301、RKGPD1 mRNA表达水平。采用气相色谱-质谱联用技术测定了YM25235中亚油酸（LA）和亚麻酸（ALA）等PUFAs的含量，并测定了YM25235在低温下的生长速率和甘油含量。构建RKHog1过表达、敲除和修复菌株。胁迫抗性分析表明，RKHog1基因的过表达增加了YM25235的甘油生物合成，增强了YM25235对冷胁迫、盐胁迫和渗透胁迫的耐受性。相反，敲除RKHog1基因减少了甘油的生物合成，抑制了YM25235对不同胁迫的耐受性。脂肪酸分析表明，过表达RKHog1基因的YM25235菌株LA和ALA含量显著升高，而敲除RKHog1基因的YM25235菌株LA和ALA含量降低。RKHog1基因过表达后，RKD12、RKMsn4、RKHisK2301和RKGPD1 mRNA的表达量在15℃时升高，而在30℃时表达量不变，RKHog1可以调节YM25235在低温下的生长适应性和PUFA含量，这可能有助于YM25235的冷适应。
The aim of this study was to investigate the role of RKHog1 in the cold adaptation of Rhodosporidium kratochvilovae strain YM25235 and elucidate the correlation of biosynthesis of polyunsaturated fatty acids (PUFAs) and glycerol with its cold adaptation. The YM25235 strain was subjected to salt, osmotic, and cold stress tolerance analyses. mRNA levels of RKhog1, Delta(12/15)-fatty acid desaturase gene (RKD12), RKMsn4, HisK2301, and RKGPD1 in YM25235 were detected by reverse transcription quantitative real-time PCR. The contents of PUFAs, such as linoleic acid (LA) and linolenic acid (ALA) was measured using a gas chromatography-mass spectrometer, followed by determination of the growth rate of YM25235 and its glycerol content at low temperature. The RKHog1 overexpression, knockout, and remediation strains were constructed. Stress resistance analysis showed that overexpression of RKHog1 gene increased the biosynthesis of glycerol and enhanced the tolerance of YM25235 to cold, salt, and osmotic stresses, respectively. Inversely, the knockout of RKHog1 gene decreased the biosynthesis of glycerol and inhibited the tolerance of YM25235 to different stresses. Fatty acid analysis showed that the overexpression of RKHog1 gene in YM25235 significantly increased the content of LA and ALA, but RKHog1 gene knockout YM25235 strain had decreased content of LA and ALA. In addition, the mRNA expression level of RKD12, RKMsn4, RKHisK2301, and RKGPD1 showed an increase at 15 degrees C after RKHog1 gene overexpression but were unchanged at 30 degrees C. RKHog1 could regulate the growth adaptability and PUFA content of YM25235 at low temperature and this could be helpful for the cold adaptation of YM25235.