| 摘要: |
| 【目的】对浙江地区采集的野生桑黄菌株进行鉴定和系统发育分析，并探讨其培养特性，为进一步开发利用桑黄菌提供科学依据。【方法】利用核糖体基因rDNA内转录间隔区（internal transcribed spacer，ITS）序列并结合形态学特征，对野生桑黄菌株（编号SA-1）进行鉴定；基于ITS1-5.8S-ITS2序列，对菌株SA-1和桑黄孔菌属的桑树桑黄、高山桑黄、锦带花桑黄、杨树桑黄、环区桑黄、小孔忍冬桑黄、暴马桑黄共8个菌株进行亲缘关系和系统发育分析；设置不同碳源、氮源、无机盐、温度和pH开展单因素试验，分析各培养因子对桑黄菌株菌丝生长的影响，并运用L9（34）正交试验对培养条件进行优化。【结果】根据菌株形态学特征和rDNA ITS序列分析，将野生菌株SA-1鉴定为桑树桑黄（Sanghuangporue sanghuang），但其与桑黄孔菌属中其他6个菌株的遗传距离较远。系统发育进化树结果显示，桑树桑黄和菌株SA-1聚为一类（bootstrap=100%），其他6个菌株聚为另一类。单因素试验确定SA-1菌丝生长的适宜培养温度为27.5 ℃，最适pH为7.0，最佳碳源为蔗糖，最佳氮源为蛋白胨，最佳无机盐为KH2PO4；经正交试验得到最适培养条件为可溶性淀粉20 g/L，蛋白胨4 g/L，KH2PO4 1 g/L，培养温度30 ℃。【结论】确定浙江地区采集的野生桑黄菌株为桑树桑黄（S.sanghuang），并初步筛选出了该菌株的适宜培养条件。 |
| 关键词: 野生桑黄 药用真菌 菌株培养 ITS鉴定 培养特性 |
| DOI： |
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| 基金项目:国家自然科学基金项目（31501815）；浙江省食用菌新品种选育项目（2016C02057-8）；杭州市重大项目（20180416A02） |
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| Isolation and cultivation of wild Sanghuangporue |
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SONG Jiling,WANG Weike,CHENG Junwen,et al
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| Abstract: |
| 【Objective】A wild Sanghuangporue strain collected in Zhejiang was identified,its phylogenetic status was analyzed,and the culture characteristics were explored to provide basis for its further development and utilization.【Method】A wild strain of Sanghuangporue (number SA-1) was identified by internal transcribed spacer (ITS) sequence of ribosome rDNA integrated with morphological characteristics.The genetic relationship among SA-1 and seven strains (S. sanghuang,S. alpinus,S.weigelae,S. vaninii,S.zonatus,S.lonicericola and S. baumii) of Sanghuangporue and phylogenetic characteristics were analyzed based on ITS1-5.8S-ITS2 sequence.The effects of different culture factors on growth of SA-1 were analyzed with single control of different factors including carbon,nitrogen,inorganic salts,temperature and pH.Then,the culture conditions were optimized by the L9（34）orthogonal test.【Result】The SA-1 strain was identified as Sanghuangporue sanghuang based on morphological characteristics and rDNA ITS analysis.The identification results showed that the SA-1 strain had further genetic distance from the other six strains of Sanghuangporue.The phylogenetic tree indicated that S. sanghuang and SA-1 were clustered into one group (bootstrap=100%),and the other six strains of Sanghuangporue were clustered into another group.The results of single factor tests showed that the optimum carbon source was sucrose,the optimum nitrogen source was peptone,the optimum incubation temperature was 27.5 ℃,the optimum pH in medium was 7.0 and the optimum inorganic salt was KH2PO4.The orthogonal test results showed that the optimum culture conditions were soluble starch 20 g/L,peptone 4 g/L,KH2PO4 1 g/L,and culture temperature 30 ℃.【Conclusion】The SA-1 collected in Zhejiang was determined as Sanghuangporue sanghuang and its optimum culture conditions were determined. |
| Key words: wild Sanghuangporus medicinal fungus strain culture ITS identification culture characteristics |
