Medical resources, seed and seedling quality of Bletilla striata

The most graded quality of medicinal resources and seed seedlings of Bletilla striata

(1) seed testing and quality grading

1. Cutting sample

The maximum mass of the seed batch is lOkg, and the test samples are taken by the freehand halving method. The samples submitted for inspection are at least 3G, and the cleanliness analysis samples are at least 0.3go.

2. Cleanliness analysis

The samples obtained from the cutting samples were screened out for 20 mesh to remove larger impurities, and then the test samples were divided into pure seeds, other plant seeds, waste seeds, residual calyx, peel fragments, pedicel and petals, mud sand and other impurities, and the weight of each component was determined. The weight of the test sample and each component is expressed as "g", four decimal places are retained, the seed purity is calculated, and if the sum of the weight of each component and the weight of the original sample increases or loses more than 5% of the weight of the original sample, it must be redone, if the increase and loss is less than 5% of the weight of the original sample, the net seed percentage is calculated.

three. Weight determination

After the purity analysis, the seeds were fully mixed evenly and divided into 4 parts by dichotomy. 1000 seeds from each net seed were randomly divided into a group and repeated for 5 times. The 1000-grain weight was recorded, the weight was kept with 4 decimal places, and the standard deviation and coefficient of variation were calculated. The coefficient of variation should not exceed 4%.

four. Germination test

The pure seeds were mixed evenly, and appropriate amount of seeds were randomly selected. The petri dish covered with two layers of wet filter paper was used as the germination bed. 100 seeds were placed in each petri dish and cultured under the condition of 30 ℃ light intensity of 2000 lx. Observe every day, replenish water, and record the number of seeds germinated on the 4th-20th day.

5. Authenticity identification

One hundred seeds were randomly selected and repeated for 3 times. the morphology, color and surface characteristics of seeds were observed under stereoscope, and the seed size was measured. Identification basis: the seeds are very small, most of them are long spindle-shaped, a few crescent-shaped, 0.62-2.69mm in length, 0.11 in diameter, 0.36mm in diameter, and the seed coat is yellow-brown, translucent and metallic luster.

6. Moisture determination

Take 0.5g of mixed white and pure seeds, put the whole seeds in a clean and constant weight aluminum box, weigh them, keep 4 decimal places and repeat 4 times. When the sample box is put into the human oven and the box temperature is maintained at (133 ±2) ℃, the calculation time begins and the sample drying time is 3 hours. Remove and cool to room temperature, then weigh. According to the formula, the water content of seed is calculated as follows: seed water content = [(sample weight before drying-sample weight after drying) / sample weight before drying] X 100%.

7. Determination of vitality

About 100 seeds were randomly selected and soaked in clean water for 4 hours at room temperature. Under the condition of 40 ℃ in 1% TTC (2jin3penol 5-triphenyltetrazolium chloride) solution, dyeing for 7 hours. Take out the seed and rinse the embryo with clean water and observe its staining. The embryo (without endosperm) is dyed red to be a viable seed.

8. Health check

The direct inspection method was used to detect the seeds infected with diseases and insect pests, and the plate culture method was used to detect the infected seeds. ① direct examination: 400 seeds were randomly selected and put on white paper or glass and examined with naked eyes. The seeds infected with diseases and insect pests were taken out, the number of seeds were calculated, and the infection rate was calculated. ② plate culture method: sterilize the petri dish and PDA medium; randomly select 100 seeds and put them in the petri dish with 15~20ml medium, 5 seeds in each petri dish, culture in a 25 ℃ incubator for 3-5 days and observe timely; pick the purer part of the fungus to another new culture medium for culture The purified fungi were stained with cotton blue stain, then observed under microscope, photographed and identified. The carrier rate and isolation rate were calculated.

(2) Classification of seed stem quality

The tubers planted for two years will be eradicated. According to the quality, the tubers are divided into I, Ⅱ and Ⅲ grades (Table 1).

Table 1 quality grade of stem

(3) grading of capsule quality

The capsule is cylindrical, 1.6-4.5cm long, 0.5-1.2cm in diameter, slightly narrowly pointed at both ends, with 6 longitudinal ribs. Take white and capsule, select clean, remove impurities, take the quality as the index, divided into I, Ⅱ, Ⅲ grade (Table 2).

Table 2 quality grade of capsule

(4) quality classification of tissue culture tubers.

The tuber culture medium was washed away and the fibrous roots were removed. According to the quality, the tubers were divided into I, Ⅱ and Ⅲ grades (Table 3).

Table 3 quality grade of tissue culture tuber

(5) the highest grade of tissue culture seedling quality.

The culture medium of tissue culture seedlings with tubers was washed out and divided into I, Ⅱ and Ⅲ grades (Table 4) according to the quality.

Table 4 quality grade of tissue culture seedlings