| The Content Variation of Natural Product Induced by Different Factor(s) | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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| Species Name: Ascochyta medicaginicola | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: PDB medium | [1] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
The live yew branches were collected from branches of T wallichiana var. mairei (Lemee et H. Lev.) L. K. Fu et Nan Li growing in the Taihang Mountain, Henan Province, China in March 2014. The hypha with PDA pieces (approximately 4 mm × 4 mm) were inoculated into 60 mL PDB culture medium in culture flask for incubation at 120 rpm and 20 to 25 ℃ for 4 d. Five milliliters of liquid culture medium containing hyphae fung was inoculated into 60 mL PDB culture medium in culture flask and cultured at 120 rpm and 20 to 25 ℃ for 8 days. The uninoculated PDB culture medium in culture flask was taken as control.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDB medium with whole culture (20-25℃ + 8 days)
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Branches | Taihang Mountain, Henan Province, China |
NP Content: 1.1215 mg/kg
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PDB medium with spent culture (20-25℃ + 8 days)
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Branches | Taihang Mountain, Henan Province, China |
NP Content: 0.936 mg/kg
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PDB medium with mycelia (20-25℃ + 8 days)
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Branches | Taihang Mountain, Henan Province, China |
NP Content: 20 mg/kg
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| Species Name: Aspergillus aculeatinus Tax-6 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: PDB medium | [2] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Fungi were isolated from the outer and inner bark of Taxus chinensis var. maire in Baiji, Anhui Province, China. The endophytic fungus was aerobically cultured in potato dextrose broth medium at 28 ℃ for one week. Plackett-Burman design: In this study, seven factors were included. For each factor, a high (+) and low (-) level was tested. Twelve runs of various levels of factors were formulated by Design-Expert version 8.0 and the response was measured according to the yield of taxol. All runs were conducted in 250 mL Erlenmeyer flasks containing 100 mL PDA liquid medium at 30 ℃ with an initial pH of 7.0. The tested factors were: sodium benzoate; salicylic acid; phenylalanine; sodium acetate; glycine; CuSO4; methyl jasmonate. After culturing for 8 days, the fungal taxol was extracted as described above.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDB medium (28℃ + 7 days)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 334.92 µg/L
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| Factor Name: PDA medium | [2] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Fungi were isolated from the outer and inner bark of Taxus chinensis var. maire in Baiji, Anhui Province, China. The endophytic fungus was aerobically cultured in potato dextrose broth medium at 28 ℃ for one week. Plackett-Burman design: In this study, seven factors were included. For each factor, a high (+) and low (-) level was tested. Twelve runs of various levels of factors were formulated by Design-Expert version 8.0 and the response was measured according to the yield of taxol. All runs were conducted in 250 mL Erlenmeyer flasks containing 100 mL PDA liquid medium at 30 ℃ with an initial pH of 7.0. The tested factors were: sodium benzoate; salicylic acid; phenylalanine; sodium acetate; glycine; CuSO4; methyl jasmonate. After culturing for 8 days, the fungal taxol was extracted as described above.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDA medium with 10, 15, 8, 25, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 492.48 µg/L
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PDA medium with 20, 5, 2, 25, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 493.81 µg/L
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PDA medium with 20, 5, 8, 15, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 468.54 µg/L
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PDA medium with 20, 15, 2, 15, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 822.1 µg/L
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PDA medium with 10, 5, 2, 15, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 50 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 355.23 µg/L
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PDA medium with 10, 15, 2, 25, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 365.25 µg/L
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PDA medium with 10, 5, 8, 25, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 50 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 384.26 µg/L
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PDA medium with 20, 15, 2, 25, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 50 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 851.25 µg/L
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PDA medium with 10, 15, 8, 15, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 50 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 404.52 µg/L
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PDA medium with 20, 5, 8, 25, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 50 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 410.45 µg/L
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PDA medium with 20, 15, 8, 15, 0.05 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 6 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 502.54 µg/L
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PDA medium with 10, 5, 2, 15, 0.15 mg/L sodium benzoate, salicylic acid, phenylalanine, glycine, CuSO4 and 10 g/L sodium acetate, 150 µmol/L methyl jasmonate model (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 305.65 µg/L
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PDA medium with 0.05 mg/L CuSO4, 5 mg/L salicylic acid and 8 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 335.28 µg/L
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PDA medium with 0.15 mg/L CuSO4, 5 mg/L salicylic acid and 10 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 476.73 µg/L
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PDA medium with 0.05 mg/L CuSO4, 15 mg/L salicylic acid and 8 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 454.64 µg/L
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PDA medium with 0.15 mg/L CuSO4, 15 mg/L salicylic acid and 8 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 282.62 µg/L
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PDA medium with 0.05 mg/L CuSO4, 10 mg/L salicylic acid and 6 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 413.73 µg/L
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PDA medium with 0.15 mg/L CuSO4, 10 mg/L salicylic acid and 6 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 547.26 µg/L
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PDA medium with 0.05 mg/L CuSO4, 10 mg/L salicylic acid and 10 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 854.9 µg/L
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PDA medium with 0.15 mg/L CuSO4, 10 mg/L salicylic acid and 10 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 843.26 µg/L
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PDA medium with 0.1 mg/L CuSO4, 5 mg/L salicylic acid and 6 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 633.38 µg/L
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PDA medium with 0.1 mg/L CuSO4, 15 mg/L salicylic acid and 6 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 338.24 µg/L
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PDA medium with 0.1 mg/L CuSO4, 5 mg/L salicylic acid and 10 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 565.72 µg/L
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PDA medium with 0.1 mg/L CuSO4, 15 mg/L salicylic acid and 10 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 828.8 µg/L
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PDA medium with 0.1 mg/L CuSO4, 10 mg/L salicylic acid and 8 g/L sodium acetate (30℃ + 8 days + PH7)
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Outer and inner bark | Baiji, Anhui Province, China |
NP Content: 1377.56 µg/L
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| Species Name: Aspergillus fumigatus strain TPF-06 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: S7 medium | [3] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Taxol-producing endophytes were isolated from different plant tissues (bark, stem, and needle) of Taxus sp. collected from Shimla, Himachal Pradesh (India). Erlenmeyer flask containing 100 mL modified S7 liquid medium was seeded with 4.79 × 104 spores per mL of the fungal isolate TPF-06 and incubated at 25 ℃ for 21 days with agitation speed at 150 rpm in an incubator shaker.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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Modified S7 liquid medium (25℃ +21 days)
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Bark; Stems; Needles | Shimla, Himachal Pradesh, India |
NP Content: 1.6 g/L
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| Species Name: Cladosporium cladosporioides strain MD-2 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: PDB medium | [4] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Samples of T. media were obtained from the campus of Huazhong University of Science and Technology (114° 31′ E, 30° 36′ N) in Hubei province in central China. Seven bark samples (0.5 × 3 cm) from yew trees were harvested and immediately transported to our laboratory and stored at 4 ℃. These endophytic fungi were cultured in 1-l Erlenmeyer flasks containing 300 ml potato dextrose liquid medium at 25 ℃ with 180 recycles/minute for 10 days.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDB medium (25℃ + 10 days)
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Inner bark | Hubei, China. |
NP Content: 800 µg/l
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| Species Name: Fusarium redolens | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: S7 Medium | [5] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Bark samples (1 × 3 cm) were harvested from the stem of relatively young T. baccata subsp. wallichiana from Bhadrewah(district Doda, Jammu & Kashmir, India) Pieces of agar block containing the mycelia mats from a 7-10-day-old culture plate of TBPJ-B were transferred to 100 ml of sterilized S-7 media and incubated at 25-28 ℃ on a rotary shaker for 5 days. This culture was used as seed culture for taxol production, where 10-20 ml of seed culture was transferred to a 2 L Erlenmeyer flask containing 500 ml of sterilized S-7 mediumand incubated at 25-28 ℃ for 21 days in the dark as the stationary culture.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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S7 Medium (25-28 degrees Celsius + 21 Days) in the dark
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bark | Bhadrewah(district Doda, Jammu & Kashmir, India) |
NP Content: 66 ng/L
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| Species Name: Penicillium aurantiogriseum NRRL 62431 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: PDB Medium | [6] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
Fungal cultures were isolated from freshly harvested Corylus avellana "Barcelona" nuts from Aurora, Oregon, USA. A 1 week-old sporulating culture on PDA was rinsed with 20 mL of sterile water containing 1 drop of Tween-20. Two mL of the spore solution with an absorbance of about 0.8 at 600 nm was added to each of 6 liters of potato dextrose broth (PDB, DIFCO). Broth cultures were shaken at 20 ℃ and 100 rpm for 2 weeks. On Day 14, when the amount of reducing sugars in the cultures was no longer detectable using glucose test strips, 100 µL of methyl jasmonate and 0.172 g/L filter-sterilized phenylalanine were added to each flask, and shaking was resumed. The cultures were harvested on day 24.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDB Medium (20 degrees Celsius + 100 rpm + 24Days) with 100 µL methyl jasmonate and 0.172 g/L filter-sterilized phenylalanine on Day 14
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nut | Aurora, Oregon, USA |
NP Content: 120 µg
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| Species Name: Pestalotiopsis microspora | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: MID Medium | [7] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
The fresh stem fragments of Taxus wallichiana Zucc., the Himalayan yew, were collected during January 1998 from Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). Several fungal endophytes were isolated from its inner bark. Each pure isolate was grown in 500 mL MID media with 1 g soytone (Difco Co) per litre for three weeks
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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500 mL MID Medium with 1 g soytone (Difco Co) per litre (21Days)
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fresh stem | Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). |
NP Content: 25.7 ng/L
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500 mL MID Medium with 1 g soytone (Difco Co) per litre (21Days)
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fresh stem | Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). |
NP Content: 167.2 ng/L
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500 mL MID Medium with 1 g soytone (Difco Co) per litre (21Days)
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fresh stem | Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). |
NP Content: 203.4 ng/L
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| Species Name: Sporormia minima | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: MID Medium | [7] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
The fresh stem fragments of Taxus wallichiana Zucc., the Himalayan yew, were collected during January 1998 from Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). One of the new isolated fungi, Sporormia minima, was cultured in 2 L MID media containing 1 g soytone per liter for three weeks
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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500 mL MID Medium with 1 g soytone (Difco Co) per litre (21Days)
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fresh stem | Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). |
NP Content: 15.7 ng/L
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| Species Name: Taxomyces andreanae | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: S7 medium | [8] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
This fungus was isolated from the surface-disinfected (80% ethanol) inner bark of one tree in an old-growth cedar forest in northern Montana. Taxomyces andreanae was cultured bytransferring hyphal tips from water agar, on which bark pieces had been cultured, onto a modified-mycological agar( S-7 medium consists of 1 g of glucose, 3 g of fructose, 6 g of sucrose, 1 g of Na+-acetate, 1 g of soytone, 1 mg of thiamine, 1 mg of biotin, 1 mg of pyridoxal, 1 mg of Ca2+-pantothenate, 3.6 of mg MgSO4, 6.5 mg of CaNO3, 1 mg of Cu(N03)2, 2.5 mg of ZnSO4, 5 mg of MnCI2, 2 mg of FeCI3, 5 mg of phenylalanine, 100 mg of Na+-benzoate, and 1 ml of 1 M KH2PO4 buffer (pH 6.8) per liter. Sugar ratio is identical to that occurring in the inner bark of Pacific yew. Modified mycological agar consists of 10 g of bacto-soytone, 40 g of glucose, 15 g of bacto-agar, 1 g of Na+-acetate, and 50 mg of sodium benzoate per liter. ). The mycelium was then successively transferred to eliminate traces of taxol or other taxanes carried over from the original tree source.Transfers of small agar plugs to broth cultures were made from mycelia that hadgrown 3 to 7 days. After this period, mycelia appeared to go into a quiescent state.Taxomyces andreanae was stored in water at 4 ℃ and grown on a semi-defined culture medium. The conidia of T. andreanae do not germinate, therefore we transferred pieces (0.5 by 0.5 cm) of agar block containing the mycelial mats to sterilized S-7 medium. Optimum conditions were as a still culture, at 25 ℃ , with a surface-to-volume ratio of 1.3 (cm2:ml). After 21 days of incubation, the culture was filtered through cheesecloth. Chlorocholine chloride (at 1 mg/ml) in the medium abolished taxol production.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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S7 medium (25℃ + 21 days)
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Phloems (Inner bark) | Montana, USA |
NP Content: 24 ng/L
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S7 medium with 1 mg/ml choline chloride (25℃ + 21 days)
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Phloems (Inner bark) | Montana, USA |
NP Content: 0 ng/L
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| Species Name: Trichothecium sp. | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: MID Medium | [7] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
The fresh stem fragments of Taxus wallichiana Zucc., the Himalayan yew, were collected during January 1998 from Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). Several fungal endophytes were isolated from its inner bark. Each pure isolate was grown in 500 mL MID media with 1 g soytone (Difco Co) per litre for three weeks
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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500 mL MID Medium with 1 g soytone (Difco Co) per litre (21Days)
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fresh stem | Shivapuri, 15 km north from Kathmandu at an altitude of 2200 m (voucher specimen KS-100 is deposited at National Herbarium Center, Godawari, Kathmandu). |
NP Content: 165.7 ng/L
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| Species Name: Tubercularia sp. strain TF5 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Factor Name: PDB Medium | [9] | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
| Experiment Detail |
The fungus used in this study was one of 45 endophytic fungi isolated from the inner bark of T. mairei obtained in Fujian province, southeast China.The endophytic fungus strain TF5 was grown in 2-lErlenmeyer asks containing 500 ml potato dextrose liquidmedium. The fungus was incubated at 25 ℃ without agitation for 21 days.
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| Factor | Part | Location | NP Content | ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
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PDB Medium (25 degrees Celsius + 21Days)
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inner bark | Fujian province, southeast China |
NP Content: 10 µl
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