Trichoderma viride|BNCC341743 |BNCC

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Trichoderma viride

Culture medium	Comprehensive PDA agar (CPDA): potato boil 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4 · 7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling liquid: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect the filtrate to a constant volume of 1.0L.
Subculture procedure	(1) Prepare a test tube containing 5~10mL of liquid medium and 2 plates; (2) open it in the biosafety cabinet, heat the tip of ampoule in the flame, quickly drop sterile water to creak it, and break it with forceps; (3) draw 0.5mL of liquid culture medium into a freeze dried ampoule, fully rehydrate and transfer the solution to the liquid test tube, mix evenly; (4) inoculate a plate with 0.2mL of suspension liquid, repeat the step to obtain two plates; ⑤ Put all the liquid test tubes and plates under the above culture conditions for cultivation, and the strains can be used when they grow.
Growth conditions	28 ℃;,3-5 days; Aerobic;
Growth characteristics	Macroscopic morphology: MEA medium, cultured at 25 ℃ for 96h, colony radius 43-45mm, colony white; The surface is flat; The texture is velvet-like; The reverse side is light yellow; No exudate is produced, no soluble pigment is produced. Microscopic morphology: On MEA medium, cultured at 25 ℃ for 10 days, the conidia stalk is stout in the main axis, with obvious separation, and secondary branches; the bottle stalk is solitary or whorled on the main axis or each, the bottle stalk is curved, columnar or middle is enlarged, the top is obviously constricted, and the size is 9.0-25 × 2.0-3.0 μm; the conidia are green, terminal, nearly spherical or oval, the long axis diameter is 2.8-5.0 μm, the aspect ratio is 1.0-1.6, and the wall is smooth. The transcribed spacer gene sequence in rDNA accepts number DQ677655, and the elongation factor gene sequence accepts number DQ672615.
Storage conditions	2-8 ℃
morphology	Small filamentous fungi, with obvious colonies, white, dense and vigorous hyphae on the integrated PDA medium, spreading to the edge of the plate, and yellowish on the back of the medium.
Sharing mode	Public welfare sharing

Trichoderma viride

Storage conditions: 2~8 ℃

No. 341743

Product format: agar slant in 14mm test tube

Validity period : growing culture, in 30 dyas

Biosafety level : 1 , handle in ultra-clean table or safety cabinet

Receiving notice: if any abnormality is found on the day of receipt, please contact the customer service within 24 hours. If it is overdue, it is deemed that the bacteria are well. The agar slant can be used directly. In principle, it can be used many times without contamination within the validity period, but viability will gradually decrease with time. Please operate in strict accordance with this instruction, otherwise the replacement of bacteria are not be available in case of aberrant growth and loss of viability.

Growth conditions :28 ℃, aerobic, integrated PDA,3-5 days. Comprehensive PDA: potato boiling solution 1.0L, glucose 20.0g,KH2PO4 3.0g,MgSO4·7H2O 1.5g, vitamin B1 trace, agar 20.0g,pH 6.0±0.2. Sterilization at 121 ℃ for 15min. Potato boiling solution: weigh 200g of peeled potato pieces, boil in boiling water for 30min, and collect filtrate to a constant volume of 1.0L.

Recovery steps:
(1)Prepare 1-2 pieces of PDA plates;
(2)sterilize the surface of test tube for the agar slant, open it in the biosafety cabinet;
(3) cut into the pieces with aseptic inoculation hoe and inoculation shovel (see attached page). the size of square pieces is 0.5 × 0.5cm2;
(4) lay flat the small pieces to the center of the agar plate;
(5)put the plates under the above culture conditions, and the strains can be used when they grow.
Recovery record: According to the recovery instructions, the results of the recovery are reported as follows:

Item	test results
viability:	good viability,in 4 days plate colonies are obvious
colony morphology:	small filamentous fungi, with obvious colonies on the integrated PDA medium, white, dense and vigorous hyphae, spreading and growing to the edge of the plate, and light yellow on the back of the medium
Conclusion:	good viability, no abnormal colony morphology, qualified