Protoplast isolation is relevant for many different applications and has been principally used in proceduresnvolving genetic manipulation. In this study, the age of mycelium, osmotic stabilizers, enzyme, incubation te...Protoplast isolation is relevant for many different applications and has been principally used in proceduresnvolving genetic manipulation. In this study, the age of mycelium, osmotic stabilizers, enzyme, incubation temperature and incubation time were evaluated in terms of their effects on protoplast yield. The young mycelia （3 d） of Tulasnella calospora were digested for 6 h at 30℃ in a mixture of 1.2 mol·L-1 MgSO_4 ＋ 10 mmoI·L-1 K2HPO4 as the osmotic stabilizer, with a 1.0% lysing enzyme and 1.5% driselase： more than 106 protoplasts mL-1 were obtained. When collected 3y density gradient centrifugation, the concentration of protoplasts can reach 107-108 protoplasts mL-1, an amount suitable enough for experiments of transformation in fungi. For every 10_5 protoplasts, about 15-25 protoplasts can egenerate after 24-36 h cultivation in a liquid medium and after 8-10 d in an agar medium. This study produced an efficient method for protoplast production, reverting them into a typical mycelia morphology using a Tulasnella calospora solate.展开更多
从曼地亚红豆杉(Taxus x media)树皮内表皮中分离得到1株产紫杉醇的内生真菌O60B1,并通过高效液相色谱法(HPLC)和质谱法(MS)分析其发酵产物,其紫杉醇产量为2.5～3.0μg/g(紫杉醇/菌丝干重),并通过形态学特征和18 S rDNA序列分析,将其初...从曼地亚红豆杉(Taxus x media)树皮内表皮中分离得到1株产紫杉醇的内生真菌O60B1,并通过高效液相色谱法(HPLC)和质谱法(MS)分析其发酵产物,其紫杉醇产量为2.5～3.0μg/g(紫杉醇/菌丝干重),并通过形态学特征和18 S rDNA序列分析,将其初步鉴定为毛霉属(Mucor sp.)真菌。展开更多
The biodiversity of arbuscular mycorrhizal fungi(AMF) was surveyed in the Kolm region of Iran in three adjacent sites, a natural stand, a 10-year-old and a 15-yearold plantation of Amygdalus scoparia. To date, there h...The biodiversity of arbuscular mycorrhizal fungi(AMF) was surveyed in the Kolm region of Iran in three adjacent sites, a natural stand, a 10-year-old and a 15-yearold plantation of Amygdalus scoparia. To date, there have been few studies of AMF biodiversity in Iran, especially in the western forests of the country. For this study, soil and root samples were taken from A. scoparia rhizosphere soil in spring and autumn. Almost half of the root length was colonized by AMF. We identified 13 AMF species belonging to Glomeraceae, Claroideoglomeraceae or Diversisporaceae. The three plantations differed in terms of soil electrical conductivity, organic C and P. Spore density was significant correlated with P concentration. Root length colonization was correlated only with soil Ca.Species diversity and richness were significantly correlated with soil N, P, organic C and spore density. AMF diversity in 15-year-old plantations was more similar to that in the natural stand than in the 10-year-old plantation. We confirmed that a 15-year-old plantation is not similar in terms of AMF colonization to natural stands. We conclude that more than 15 years are required for AMF colonization of plantations to resemble that of natural stands.展开更多
基金supported by the National Key Project of Scientific and Technical Supporting Programs funded by the Ministry of Science & Technology, China (No. 2012BAC01B05-3)
文摘Protoplast isolation is relevant for many different applications and has been principally used in proceduresnvolving genetic manipulation. In this study, the age of mycelium, osmotic stabilizers, enzyme, incubation temperature and incubation time were evaluated in terms of their effects on protoplast yield. The young mycelia （3 d） of Tulasnella calospora were digested for 6 h at 30℃ in a mixture of 1.2 mol·L-1 MgSO_4 ＋ 10 mmoI·L-1 K2HPO4 as the osmotic stabilizer, with a 1.0% lysing enzyme and 1.5% driselase： more than 106 protoplasts mL-1 were obtained. When collected 3y density gradient centrifugation, the concentration of protoplasts can reach 107-108 protoplasts mL-1, an amount suitable enough for experiments of transformation in fungi. For every 10_5 protoplasts, about 15-25 protoplasts can egenerate after 24-36 h cultivation in a liquid medium and after 8-10 d in an agar medium. This study produced an efficient method for protoplast production, reverting them into a typical mycelia morphology using a Tulasnella calospora solate.
文摘从曼地亚红豆杉(Taxus x media)树皮内表皮中分离得到1株产紫杉醇的内生真菌O60B1,并通过高效液相色谱法(HPLC)和质谱法(MS)分析其发酵产物,其紫杉醇产量为2.5～3.0μg/g(紫杉醇/菌丝干重),并通过形态学特征和18 S rDNA序列分析,将其初步鉴定为毛霉属(Mucor sp.)真菌。
基金funded by the Ilam University(Grant No.32.1164.)
文摘The biodiversity of arbuscular mycorrhizal fungi(AMF) was surveyed in the Kolm region of Iran in three adjacent sites, a natural stand, a 10-year-old and a 15-yearold plantation of Amygdalus scoparia. To date, there have been few studies of AMF biodiversity in Iran, especially in the western forests of the country. For this study, soil and root samples were taken from A. scoparia rhizosphere soil in spring and autumn. Almost half of the root length was colonized by AMF. We identified 13 AMF species belonging to Glomeraceae, Claroideoglomeraceae or Diversisporaceae. The three plantations differed in terms of soil electrical conductivity, organic C and P. Spore density was significant correlated with P concentration. Root length colonization was correlated only with soil Ca.Species diversity and richness were significantly correlated with soil N, P, organic C and spore density. AMF diversity in 15-year-old plantations was more similar to that in the natural stand than in the 10-year-old plantation. We confirmed that a 15-year-old plantation is not similar in terms of AMF colonization to natural stands. We conclude that more than 15 years are required for AMF colonization of plantations to resemble that of natural stands.
