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基于转录组分析高温抑制广东虫草原基形成的调控网络
Dissecting the regulation network of high temperature inhibiting primordial formation of Cordyceps guangdongensis based on transcriptomic data
, ({{custom_author.role_en}}), {{javascript:window.custom_author_en_index++;}}在大型真菌原基形成和子实体发育的过程中,温度是一个极其关键的因素,高温显著影响多种食用菌原基的形成和子实体的品质。广东虫草是中国华南地区特有的虫草类新食品原料,其子实体富含多种活性成分和营养成分,且能够进行较大规模人工栽培。然而,温度对广东虫草原基形成的影响及调控机制尚不清楚。本研究通过不同温度处理不同时间段,发现29℃处理3d抑制了广东虫草原基的形成;对29℃处理3d处理前后的菌丝阶段（CK）和原基阶段（CK-P和HT-P）样品进行转录组测序分析,高温处理后原基阶段的两个组中发现1 682个差异表达基因,其中1 015个上调表达和667个下调表达。在碳水化合物代谢途径中,多个糖酵解和三羧酸循环及葡聚糖和海藻糖合成相关基因在高温处理后呈现下调表达;在原基样品中,多个热激蛋白基因（Hsp10、Hsp23、DnaJ、Hsp70、Hsp90和Hsp98）和转录因子C2H2转录水平显著上调表达。本研究结果基于分子水平揭示了高温影响原基形成过程中能量代谢和相关基因的差异表达,为后续利用广东虫草抗逆相关基因资源培育新品种奠定了重要的基础。
During the process of primordial formation and fruiting body (FB) development in macro-fungi, temperature is an extremely crucial factor, and high temperature significantly affects primordial formation and FB quality. The fruiting bodies of Cordyceps guangdongensis, a new and special food resource of Cordyceps-like fungi in South China, have abundant bioactive and nutritional ingredients, and are cultivatable in a large scale. Nevertheless, the effect and regulatory mechanisms of temperature on its primordial formation remain less-known. In this study, we found that 3d treatment under 29°C obviously inhibited the primordial formation of T. guangdongense using different period treatments of different temperatures. After treatment, the samples of mycelium (CK) and primordium (CK-P and HT-P) stages were used for transcriptome sequencing. In the comparison of HT-P vs CK-P, 1 682 differentially expressed genes (DEGs) were gained. Among them, 1 015 genes were markedly up-regulated, and 667 genes were obviously down-regulated. In terms of the carbohydrate metabolic pathway, several genes involved in glycolysis, TCA cycle, glucan and trehalose synthesis were significantly down-regulated. In additions, many genes encoding heat shock protein (Hsp10, Hsp23, DnaJ, Hsp70, Hsp90 and Hsp98) and transcription factor C2H2 were markedly up-regulated in primordia. At molecular level, the results of this study revealed the differential expression of energy metabolism and the relative genes during the process of high temperature affecting primordial formation, laying an important foundation for breeding new varieties of C. guangdongensis using the stress-related genes in the future.
广东虫草 / 高温胁迫 / 原基形成 / 差异表达基因 / 热激蛋白 / 转录因子 {{custom_keyword}} /
Cordyceps guangdongensis / high temperature stress / primordial formation / differentially expressed genes / heat shock protein / transcription factor {{custom_keyword}} /
图3 剑孢新赤壳 (HMAS 290889)A-C：自然基物上的子囊壳;D,E：25 ℃培养7 d的菌落形态 (D：PDA;E：SNA);F：子囊壳纵切面结构;G-I：子囊及子囊孢子;J-L：子囊孢子;M,N：分生孢子梗和小型分生孢子;O：小型分生孢子;P-S：大型分生孢子. 标尺：A-C=1 mm;F=50 μm;G-S=10 μm Fig. 3 Neocosmospora protoensiformis (HMAS 290889). A-C: Ascomata on natural substratum; D, E: Colonies after 7 d at 25 °C (D: PDA; E: SNA); F: Median section of an ascoma; G-I: Asci with ascospores; J-L: Ascospores; M, N: Conidiophores and microconidia; O: Microconidia; P-S: Macroconidia. Bars: A-C=1 mm; F=50 μm; G-S=10 μm. |
图5 瘤顶赤壳 (HMAS 290890)A-C：自然基物上的子囊壳;D,E：25 ℃培养14 d的菌落形态 (D：PDA;E：SNA);F：子囊壳纵切面结构;G-K：分生孢子梗和分生孢子;L：厚垣孢子. 标尺：A-C=1 mm;F=50 μm;G-L=10 μm Fig. 5 Tumenectria laetidisca (HMAS 290890). A-C: Ascomata on natural substratum; D, E: Colonies after 14 d at 25 °C (D: PDA; E: SNA); F: Median section of an ascoma; G-K: Conidiophores and conidia; L: Chlamydospores. Bars: A-C=1 mm; F=50 μm; G-L=10 μm. |
图6 基于ITS和LSU序列的MP树粗体显示5个中国新记录种的系统发育位置,MPBP大于50% (左)、BIPP大于90% (右)标注于分支节点上 Fig. 6 Maximum parsimony phylogram reconstructed from the combined sequences of ITS and LSU. the phylogenetic position of the five Nectriaceae species new to China. MPBP above 50% (left) showing and BIPP above 90% (right) are given respectively. |
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