Identification of suitable reference genes during the formation of chlamydospores in Clonostachys rosea 67-1.
Identification of suitable reference genes during the formation of chlamydospores in Clonostachys rosea 67-1.
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DOI：
10.1002/mbo3.505
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发表时间：
2017-10
期刊：
影响因子：
3.4
通讯作者：
Sun M
中科院分区：
文献类型：
--
作者：
Zhang J;Sun Z;Li S;Sun M
玫瑰粘帚霉是一种很有潜力的生防真菌，在特定条件下能产生高抗性厚垣孢子。为了研究与厚垣孢子形成相关的基因，我们鉴定了可靠的参考基因，用于定量孢子形成期间玫瑰色念珠菌67 - 1中的基因表达。在这项研究中，9个参考基因，肌动蛋白（ACT），延伸因子1（EF 1），甘油醛-3-磷酸脱氢酶（GAPDH），组蛋白（HIS），RNA聚合酶II CTD磷酸酶Fcp 1（RPP），琥珀酸半醛脱氢酶（SSD），TATA结合蛋白（TBP），泛素（UBQ）和泛素缀合酶（UCE），从67 - 1，并使用逆转录定量PCR测定它们在厚垣孢子形成期间的表达稳定性，并使用软件geNorm、Normandum和BestKeeper进行评估。候选基因的Ct值在19.9 ~ 29.7之间，其中HIS、ACT和SSD表达量较高。统计分析表明，ACT和SSD在不同培养条件下表达最稳定，而UBQ和GAPDH在不同培养条件下表达差异较大。计算成对变异值表明，精确定量需要两个参考基因。最后，选择ACT和SSD以使玫瑰色念珠菌67 - 1中厚垣孢子产生期间的基因表达标准化。据我们所知，这是SSD作为参考基因的首次报道。本研究将有助于准确定量厚垣孢子形成过程中差异表达的基因，并有助于研究厚垣孢子形成的分子机制。
Clonostachys rosea is a potential biocontrol fungus that can produce highly resistant chlamydospores under specific conditions. To investigate the genes related to chlamydospore formation, we identified reliable reference genes for quantification of gene expression in C. rosea 67‐1 during sporulation. In this study, nine reference genes, actin (ACT), elongation factor 1 (EF1), glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH), histone (HIS), RNA polymerase II CTD phosphatase Fcp1 (RPP), succinate‐semialdehyde dehydrogenase (SSD), TATA‐binding protein (TBP), ubiquitin (UBQ), and ubiquitin‐conjugating enzyme (UCE), were selected and cloned from 67‐1, and their expression stability during chlamydospore formation was determined using reverse transcription quantitative PCR and assessed using the software geNorm, NormFinder and BestKeeper. The Ct values of the candidates ranged from 19.9 to 29.7, among which HIS,ACT and SSD exhibited high expression levels. The statistical analysis showed that ACT and SSD were most stably expressed, while UBQ and GAPDH showed relatively large variations under different culture conditions. Calculation of pairwise variation value indicated that two reference genes were required for precise quantification. Finally, ACT and SSD were selected to normalize gene expression during chlamydospore production in C. rosea 67‐1. To the best of our knowledge, this is the first report of SSD as a reference gene. This study will facilitate the accurate quantification of differentially expressed genes during the generation of chlamydospores and contribute to the investigation of the molecular mechanism underlying chlamydospore formation in C. rosea.
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影响因子：
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作者：
Li QQ;Skinner J;Bennett JE
通讯作者：
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影响因子：
3.7
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影响因子：
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