Strain of Tulasnella calospora QS104 and application and method for promoting growth of paphiopedilumm aseptic seedlings
A technology of QS104 and gelatinous bacteria, applied in the direction of microorganism-based methods, botany equipment and methods, biochemical equipment and methods, etc., can solve the problems of low survival rate of transplanting, restriction of artificial breeding of Paphiopedilum, and slow growth of aseptic seedlings And other issues
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Embodiment 1
[0048] 1. Observation and isolation of mycorrhizal fungi
[0049] Material: P. armeniacum (P. armeniacum) is a terrestrial or semi-epiphytic plant, produced in western Yunnan (Bijiang, Hushui). Born in limestone soil at an altitude of 1400-2100 meters or on rocky and well-drained grass slopes, flowering period is February-April.
[0050] Laser confocal observation of the root structure to determine the root material: WGA-AF (wheat germ agglutinin-alexa fluor conjugate) 488 (Molecular Probes) can specifically stain the chitin of the hyphae, and the root material was fixed in absolute ethanol for more than 24 hours , cut into thin slices of about 1 mm by hand, placed in 10% KOH solution overnight, and placed the material in the dye solution (10mg / ml WGA-AF 488, 0.02% Tween20 dissolved in 1×PBS (137mM NaCl, 27mM KCl, 100mM Na2HPO4, 2mM K2HPO4, pH 7.4) stained for 2h, vacuumed once every 15min during the staining period, then the material was placed in 1 × PBS solution and rinsed...
Embodiment 2
[0101] Example 2 Co-cultivation of Coomenophora mesorium QS104 and Paphiopedilum apricot aseptic seedlings
[0102] 1. Materials and methods:
[0103] The tested seedlings of Paphiopedilum apricot were 9-month-old uniform-sized sterile seedlings (plant height 2-3cm, leaves 4-5, roots 2-4, root length about 1cm). The tested strain is an endophytic fungus isolated from P. mesoporium QS104 (preservation number CGMCC NO.17777).
[0104] The co-cultivation medium was DE medium.
[0105] DE medium formula: CaCl 2 1.0mmol / L, MgSO 4 0.5mmol / L, K 2 SO 4 1.0mmol / L, KH 2 PO 4 0.4mmol / L, FeSO 4 100μmol / L, H 3 BO 4 25μmol / L, MnCl 2 33μmol / L, ZnSO 4 2.8 μmol / L, NaMoO 4 1.0 μmol / L, Na 2 EDTA 140μmol / L, yeast extract 1.0g / L, agar 8.5g / L, pH 6.0.
[0106] Co-cultivation method: insert Paphiopedilum seedlings into orchid tissue culture bottles filled with culture medium (volume 600mL, bottle mouth inner diameter 4cm, culture medium 120mL / bottle) in a sterile operating tabl...
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