Comparative experiment on liquid strain substitute cultivation of six strains of Lentinus edodes

In recent years, the production of Lentinus edodes in China has developed rapidly, especially the substitute cultivation of Lentinus edodes. The traditional solid seed production technology of Lentinus edodes can be extended to the production of cultivation bags after the three-stage cultivation of mother seed, original seed and cultivated species. The development of Lentinus edodes is greatly restricted by its great labor intensity, complicated technology, high contamination rate of miscellaneous bacteria, long growth cycle and so on. In this experiment, six liquid strains of Lentinus edodes were cultured and cultivated, in order to provide a theoretical basis for the popularization and application of liquid strains of Lentinus edodes.

1 materials and methods

1.1 strains L135 (Huazhong Agricultural University), L939 (Huazhong Agricultural University), L241-4 (Laboratory of Edible Fungi of Xinyang Agricultural College), L103 (Xixia County Nongfeng Fungi Industry Company), L9608 (Henan Academy of Agricultural Sciences), L9015 (Institute of Biology, Henan Academy of Sciences).

① liquid medium (CYM): peptone 2g, yeast extract 2g, magnesium sulfate 0.5g, potassium dihydrogen phosphate 0.46g, dipotassium hydrogen phosphate 1.0g, glucose 20g, Agar 20g, distilled water 1000mL. ② culture medium: sawdust 77%, wheat bran 10%, rice bran 10%, sucrose 0.7%, potassium dihydrogen phosphate 0.3%, gypsum 1%, quicklime 1%. The water content is 55% and 60%.

1.3 liquid culture method: different strains of Lentinus edodes were cultured in (25 ±1) ℃ for 9 days, and then transferred into liquid medium after the mycelium was full of slant. Each bottle was inoculated with 3 pieces of hyphae per bottle, and 3 bottles of each treatment (formula) were repeated. 25 ℃ static culture for 48 hours, then shaking culture for 6 days at rotational speed of 120r/min and temperature of (25 ±1) ℃. When many small and dense mycelium balls were produced in the bottle, the culture was terminated.

Determination of nutritional composition of liquid mycelium six Lentinus edodes strains were inserted into liquid medium, each bottle of 100mL culture medium was connected with 0.5cm mycelium block and repeated in 5 bottles. After the culture was terminated, the fermented culture medium was filtered with 60 mesh sieve, and the mycelium and filtrate of each strain were collected. The mycelium was washed with distilled water twice and dried to constant weight at 60 ℃. The dry weight of mycelium was measured by electronic balance. The pH value was determined by precision pH test paper.

1.5 comparison of culture characteristics of liquid bacteria in this experiment, the specification of cultivation bag was 55cm × 15cm × 0.045cm, each bag was filled with dry material 1.5kg, and then inoculated under the condition of 20 ℃ and 25 ℃, 20 bags for each variety. When the hyphae began to germinate, the mycelium growth rate was measured with a ruler. After the mycelium was full, the mycelium was moved into the mushroom room to remove the bag to produce mushroom, and the characteristics of mushroom production were compared.

2 results and analysis

2.1 determination of mycelium biomass and pH value of liquid strains of different Lentinus edodes strains

Six strains of Lentinus edodes were put into the liquid medium of the same formula and each bottle of 100mL culture medium was repeated for 5 times. After the end of culture, the dry weight and initial and final pH of mycelia among different strains of Lentinus edodes were determined. The results are shown in Table 1.

Table 1 Biomass (mg/mL) and pH among different strains of Lentinus edodes

The initial pH of mycelium dry weight of variety ends the change of pH value.

L960816.764.21.8

L241-416.464.21.8

L9399.763.92.1

L1359.963.92.1

L90158.964.21.8

L1037.863.92.1

It can be seen from Table 1 that the mycelial dry weight of 6 Lentinus edodes strains is higher than that of L9608 and L241-4. The results showed that the two strains decomposed and utilized various nutrients quickly in liquid medium, which was beneficial to the improvement of mycelium absorption, growth and vitality, and was suitable for liquid culture. The mycelium dry weight of the other four strains was lower, which indicated that the strain was not suitable for liquid culture. The pH values of 6 strains of Lentinus edodes changed little.

2.2 the growth of different mycelia in the material bag the liquid strains of 6 Lentinus edodes strains were inoculated on the cultivation materials of the same formula, the cultivation bags of the same size and the same material in each bag were used and cultured under the condition of 20 ~ 25 ℃ after inoculation. with 10 days as the interval, the mycelium growth rate of each bag was measured with ruler and the average value was calculated. The mycelium growth is shown in Table 2.

Table 2 mycelial growth of different strains in bacterial bags (mm)

Mycelial growth L9608L9015L103L939L241-4L135

Mycelium growth rate 32.1928.7227.1429.3232.3031.28

36.8727.6328.3228.2637.4329.33

31.6435.2829.2434.3430.3027.29

Average 33.5730.5528.2330.6433.3429.30

Note: the above values are measured on the 10th day.

As can be seen from Table 2, when the six strains of Lentinus edodes were used in cultivation, the resistance of L103, L135, L939 and L9015 was relatively weak and the mycelium grew slowly. The mycelium growth rate of L9608 and L241-4 is relatively fast, and the pollution is relatively low. The mycelium growth of L9608 was the fastest, and the average value of 10 days was 33.57mm. The mycelium growth of L103 was the slowest, indicating that the mycelium viability of different strains was different.

2.3 the cultivation test results of different strains are shown in Table 3.

Table 3 characteristics of mushroom emergence of different strains (unit: G,%)

Characteristics of mushroom emergence early and late mushroom cover size stalk long fruiting body color total yield bioefficiency ranking

L241-4 single medium-large and medium yellowish brown 2395279.422

L9608 single medium and short light brown 2395579.831

L939 single medium short brown 2060168.671

L135 early morning single / medium tea brown 2095869.864

L9015 Zhongzao single / clump short light brown 2142671.423

L103 morning single size short brown 1972565.766

Average 2176972.33

It can be seen from Table 3 that L241-4, L939 and L9608 among the 6 Lentinus edodes strains come out late and belong to late-maturing strains. The highest yield was L9608, followed by L241-4, and the biological efficiency was L9608 > L241-4 > L9015 > L135 > L939 > L103.

3 discussion

In this experiment, through the analysis of the liquid cultivation characteristics of 6 Lentinus edodes strains, it can be seen that among the 6 strains, L9608 and L241-4 have certain advantages in mycelial activity, growth potential and impurity resistance, which are suitable for liquid strain substitute cultivation. Further experiments are needed to compare the culture characteristics, mushroom production characteristics and yield of the same strain under different temperature and other ecological factors.

Note: I am deeply grateful for the strong support from Professor Lu Zuozhou of Huazhong Agricultural University in the course of the experiment.