WO2019091497A1 - Earthworm juvenile individual and group quality evaluation methods - Google Patents

Abstract

Provided is an earthworm juvenile individual quality evaluation method, comprising the following steps: observing the integrity, body colour and reproductive ring of an earthworm individual, and measuring the weight, body length, reproductive ring diameter and vitality of the earthworm individual; if the earthworm individual is complete, the back is brown to purple-grey, the abdomen is light yellow-brown, the reproductive ring is not obvious, the weight is ≥4.0g, the body length is ≥9.0cm, the reproductive ring diameter is ≥4.5mm and the vitality is ≥35.0s, determining that the earthworm individual is an earthworm juvenile which is up to standard. Also provided is an earthworm juvenile group quality evaluation method, comprising the following steps: randomly collecting several earthworm individuals, measuring the weight, body length, reproductive ring diameter and vitality of the earthworm individuals, calculating an average value X1 and a standard deviation SV1 for the earthworm individual weight, an average value X2 and a standard deviation SV2 for the earthworm individual body length, an average value X3 and a standard deviation SV3 for the earthworm individual reproductive ring diameter and an average value X4 and a standard deviation SV4 for the earthworm individual vitality, and according to the average values and standard deviations for the weight, body length, reproductive ring diameter and vitality of the earthworm individuals, respectively calculating coefficients of variation CV1, CV2, CV3 and CV4 for the earthworm individual weight, body length, reproductive ring diameter and vitality; if X1≥5.5g, CV1≤0.28, X2≥11.0cm, CV2≤0.2, X3≥5.0mm, CV3≤0.16, X4≥42s and CV4≤0.15, determining that the earthworm juvenile group is an earthworm juvenile group which is up to standard.

Description

Individual and population quality evaluation method for Agrocybe aegerita Technical field:

The invention belongs to the technical field of cockroach breeding and Chinese medicinal materials development, and particularly relates to an individual and population quality evaluation method for radix chinensis seedlings.

Background technique:

Earth dragon, also known as cockroach, is a common terrestrial animal. The earthworm is cold and salty. The main effects are clearing heat and calming, collaterals, asthma, and diuresis. Studies have shown that Dilong has the effects of lowering blood pressure, relieving heat, calming stagnation, anticoagulation, antithrombotic, anticancer and enhancing immune function. At present, the medicinal materials of Dilong in China are mainly “Guangdi Long” and “Shanghai Dilong”. Among them, Guangdilong is a Guangdong medicinal material and is recognized as the best quality in the medicinal materials industry.

As the health needs of human medicine continue to grow and change, the demand for Chinese medicine resources is growing. Since most of the Chinese herbal medicines in China come from the wild, with the reduction of wild resources, the artificial breeding of Guangdilong will become the main source. Therefore, screening out the wide-ranging seedlings in line with the specifications is the primary link of the artificial breeding of the earthworms, which is conducive to the growth and reproduction of the earthworms, and also provides quality assurance for the production of the medicinal herbs.

Summary of the invention:

The object of the present invention is to overcome the deficiencies in the prior art and to provide an individual and population quality evaluation method for the seedlings.

A first object of the present invention is to provide a method for evaluating individual quality of A. glabra seedlings, which comprises the steps of: observing the integrity, body color and reproductive ring zone of Guangdilong, and measuring the individual weight of Guangdilong. Body length, reproductive ring diameter and vitality; if the Guanglong dragon is intact, the back is brown to purple gray, the abdomen is light yellow-brown, the reproductive ring is not obvious, and at the same time it satisfies the weight of Guangdi dragon ≥4.0g, body length ≥ 9.0cm, the diameter of the reproductive ring is ≥4.5mm and the vitality is ≥35.0s, then it is judged that the Guanglong individual is a qualified Guangdi Longyan seedling.

The body length of the genus of the genus is the length of the reproductive ring to the tail.

Preferably, the measuring body length of the Phyllostachys pubescens is specifically to place the phoenix dragon in a volume fraction of 50% alcohol, and to anesthesia until the body is in a relaxed state, measuring the length of the reproductive ring to the tail.

Preferably, the measuring the diameter of the individual reproductive ring of the typhoon is specifically measuring the diameter of the reproductive ring in the quiet state of the typhoon.

Preferably, the measuring the activity of the Phyllostachys pubescens is specifically to place the phoenix dragon in a volume fraction of 50% alcohol, and record the time required for the anesthesia to be in a relaxed state.

Preferably, the guangdilong individual is completely intact and the individual is intact and intact.

Before the evaluation of the quality of the seeds of the cultivar, the cultivar was tested for DNA molecular identification.

The molecular identification of the typhoon DNA: extracting the genomic DNA of the genus, and designing primers for PCR amplification according to the mitochondrial COI and 16SrRNA gene sequences of the typhoon in the gene bank, PCR product electrophoresis detection, and analysis of the band amplification results. The sequence of the gene was sequenced, and the sequenced results were spliced and then searched by BLAST (Basic Local Alignment Search) similarity search to find the matched sequence. The similarity between the mitochondrial COI and 16SrRNA gene sequences in the test sample and the gene bank was analyzed to confirm whether the sample was Wideland dragon variety.

A second object of the present invention is to provide a method for evaluating the quality of the seedlings of the genus Amaranthus chinensis, which comprises the following steps: randomly collecting a plurality of genus genus, measuring the individual weight, body length and reproduction of the genus With the diameter and vigor of the ring, the average value X1 and standard deviation SV1 of the collected individual weight of the genus, and the mean value X2 and standard deviation SV2 of the body length of the genus, and the average diameter of the genital ring of the genus X3 and standard deviation SV3, mean value X4 and standard deviation SV4 of Guangdilong individual vitality, according to the average and standard deviation of the individual weight, body length, reproductive band diameter and vitality of Guangdilong, respectively, calculate the Guanglong individual Discrete coefficients CV1, CV2, CV3 and CV4 of weight, body length, reproductive ring diameter and vitality; if simultaneously satisfy X1≥5.5g, CV1≤0.28, X2≥11.0cm, CV2≤0.2, X3≥5.0mm, CV3≤ 0.16, X4≥42s and CV4≤0.15, it is judged that the population of the long-range seedlings is a qualified population of Longjing seedlings.

The body length of the genus of the genus is the length of the reproductive ring to the tail.

Preferably, the measuring body length of the Phyllostachys pubescens is specifically to place the phoenix dragon in a volume fraction of 50% alcohol, and to anesthesia until the body is in a relaxed state, measuring the length of the reproductive ring to the tail.

Preferably, the measuring the diameter of the individual reproductive ring of the typhoon is specifically measuring the diameter of the reproductive ring in the quiet state of the typhoon.

Preferably, the measuring the activity of the Phyllostachys pubescens is specifically to place the phoenix dragon in a volume fraction of 50% alcohol, and record the time required for the anesthesia to be in a relaxed state.

The invention has the beneficial effects that the evaluation method of the invention is scientific, simple and convenient, and requires less experimental instruments. The evaluation method of the invention can be used for harvesting, placing and breeding of the seedlings of Guangdilong, and promoting the growth and reproduction of the earthworm.

BRIEF DESCRIPTION OF THE DRAWINGS:

Figure 1 is the CO I sequence of the typhoon (Ginseng).

Detailed ways:

The following examples are intended to further illustrate the invention and not to limit the invention.

Example 1: Identification of DNA of Guanglonglong DNA

Random samples were collected from the Guangdilong (Shenhuan) breeding base in Luchuan County, Guangxi, and the genomic DNA of Dilong was extracted. The primers were designed for PCR amplification based on the mitochondrial COI gene sequence of the gene in the gene bank. The PCR products were sequenced. After the obtained gene sequence results are spliced (as shown in Figure 1, as shown in SEQ ID NO. 1), the BLAST (Basic Local Alignment Search) similarity search is performed to find the matched sequence, and the detection sample and the gene pool are analyzed. The sequence similarity of the mitochondrial COI gene of Guangdilong, combined with morphological analysis, determined that the sample was collected from Guangdilong.

Example 2:

A random sample of 50 samples, numbered 1 to 50, was collected from the breeding ground of Guangdilong (Shenhuan) in Luchuan County. All the collected Guangdi dragons have intact organs and the individual intact without damage, the back is brown to purple gray, the abdomen is light yellow-brown, and the reproductive ring is not obvious. After morphological identification and DNA molecular identification, they are all varieties of Guangdi.

1, Guangdi dragon weight measurement

The weight of 50 samples was weighed using an electronic balance, repeated 3 times, and averaged. The statistical results are shown in Table 1 and Table 2 below:

Table 1: Results of weight measurement of Guangdi Longyan seedlings (unit: g)

Numbering	weight	Numbering	weight	Numbering	weight	Numbering	weight	Numbering	weight
1	4.2	11	8.2	twenty one	5.8	31	6.6	41	3.5
2	3.1	12	6.7	twenty two	7.5	32	8.0	42	7.1
3	2.2	13	6.5	twenty three	4.3	33	5.0	43	6.4
4	4.6	14	3.5	twenty four	9.9	34	6.0	44	4.7
5	4.4	15	4.6	25	6.3	35	5.4	45	5.8
6	7.7	16	6.0	26	4.3	36	6.3	46	3.6
7	6.8	17	6.3	27	6.3	37	5.3	47	5.4
8	6.7	18	5.1	28	6.8	38	5.2	48	4.8
9	3.4	19	6.9	29	2.6	39	7.3	49	5.8
10	5.8	20	5.9	30	7.4	40	5.9	50	6.4

Table 2: The number of weight range of Guangdi Longyan seedlings

Weight range (unit: g)	Quantity
<3.0	2
[3.0-4.0)	5
[4.00-5.0)	8
[5.00-6.0)	12
[6.00-7.0)	15
[7.00-8.0)	5
[8.0-9.0)	2
≥9.0	1

Note: "[" stands for greater than or equal to, ")" stands for less than

2, Guangdi dragon reproductive ring belt diameter measurement

Use the vernier caliper to measure the diameter of the reproductive ring in the quiet state of the terrapin, repeat 3 times, and take the average. The statistical results are shown in Table 3 and Table 4 below:

Table 3: Results of the measurement of the diameter of the reproductive ring of Guangdi Longjing (live-quiet) (unit: mm)

Numbering	diameter	Numbering	diameter	Numbering	diameter	Numbering	diameter	Numbering	diameter
1	4.8	11	6.2	twenty one	5.5	31	5.7	41	4.3
2	4.2	12	6.6	twenty two	5.8	32	6.3	42	6.7
3	5.2	13	5.9	twenty three	4.4	33	5.2	43	6.3
4	5.8	14	4.9	twenty four	5.9	34	5.5	44	5.8
5	5.7	15	6.2	25	4.1	35	5.3	45	5.2
6	7.4	16	5.6	26	5.6	36	4.7	46	5.1
7	5.3	17	5.7	27	5.5	37	5.1	47	5.3
8	7.2	18	5.1	28	6.1	38	4.4	48	5.0
9	3.1	19	5.2	29	4.7	39	6.1	49	4.7
10	5.3	20	4.6	30	6.8	40	5.1	50	5.4

Table 4: Frequency range of the diameter of the reproductive ring zone of the radix

Note: "[" stands for greater than or equal to, ")" stands for less than

3, Guangdi dragon body length measurement

Place the Guanglong dragon in a volume fraction of 50% alcohol, and wait until it is anesthetized until the body is in a relaxed state. Use a ruler to measure the length of the reproductive ring to the tail, repeat 3 times, and take the average. The statistical results are shown in Table 5 and Table below. 6:

Table 5: Determination of body length of 50% alcohol treated radix rehmanniae (reproductive annulus to tail) (unit: cm)

Numbering	length	Numbering	length	Numbering	length	Numbering	length	Numbering	length
1	13.2	11	12.7	twenty one	9.5	31	12.2	41	8.5
2	10.2	12	11.2	twenty two	13.2	32	13.5	42	11.5
3	7.5	13	12.5	twenty three	10.8	33	10.1	43	11.8
4	8.5	14	9.2	twenty four	16.9	34	11.3	44	10.6
5	10.2	15	10.7	25	11.2	35	13.4	45	13.0
6	12.3	16	12.0	26	9.4	36	9.8	46	6.8
7	15.3	17	11.6	27	12.9	37	11.5	47	11.1
8	13.5	18	9.5	28	11.2	38	9.6	48	12.7
9	7.3	19	11.1	29	6.8	39	15.5	49	11.3

10

13.4

20

14.1

30

12.6

40

11.4

50

12.4

Table 6: Broad-range frequency of Longjing seedlings

Note: "[" stands for greater than or equal to, ")" stands for less than

4. Guangdilong vitality evaluation

The ground dragon was placed in a volume fraction of 50% alcohol until it was anesthetized until the body was in a relaxed state. The time consumption of each sample was recorded using a stopwatch. The statistical results are shown in Tables 7 and 8 below:

Table 7: Results of the determination of the activity of the radix seedlings (50% alcohol) (unit: s)

Table 8: Vitality of Guangdi Longyan Seedlings

Vitality (unit: s)	Frequency
Less than 34.0	3
[34.0-37.0)	4
[37.0-40.0)	9
[40.0-43.0)	12
[43.0-46.00)	9
[46.0-49.0)	6
[49.0-52.0)	4
Greater than 52.0	3

Note: "[" stands for greater than or equal to, ")" stands for less than

5. Quality evaluation of the individual and population of Guangdilong:

1) Quality evaluation results of Guangdilong individual:

According to the results of the measurements of the scale weight, reproductive ring diameter, body length and vigor of Table 1, Table 3, Table 5 and Table 7, combined with the frequency/number of Table 2, Table 4, Table 6 and Table 8, 50 The quality of each sample was evaluated. 36 Guangdilong individuals numbered 1, 6-8, 10-13, 15-22, 24, 26-28, 30-34, 36, 37, 39, 40, 42-45, 47, 49, 50 At the same time, the individual weights of Guangdilong ≥4.0g, body length ≥9.0cm, reproductive ring diameter ≥4.5mm and vitality ≥35.0s are satisfied, then the 36 Guangdilong individuals are judged as qualified Guangdilong individuals (see Table 9). ).

Table 9: Quality evaluation results of Guangdi Longyan seedlings

Numbering	whether it is passed	Numbering	whether it is passed	Numbering	whether it is passed	Numbering	whether it is passed	Numbering	whether it is passed
1	qualified	11	qualified	twenty one	qualified	31	qualified	41	-

2	-	12	qualified	twenty two	qualified	32	qualified	42	qualified
3	-	13	qualified	twenty three	-	33	qualified	43	qualified
4	-	14	-	twenty four	qualified	34	qualified	44	qualified
5	-	15	qualified	25	-	35	-	45	qualified
6	qualified	16	qualified	26	qualified	36	qualified	46	-
7	qualified	17	qualified	27	qualified	37	qualified	47	qualified
8	qualified	18	qualified	28	qualified	38	-	48	-
9	-	19	qualified	29	-	39	qualified	49	qualified
10	qualified	20	qualified	30	qualified	40	qualified	50	qualified

According to the results of the measurements of the weight of the genus Dragon, the diameter of the reproductive ring, the length of the body and the vitality of Table 1, Table 3, Table 5 and Table 7, the average value of the individual weights of 50 cultivar X1 = 5.7 g, standard deviation SV1 =1.53, discrete coefficient CV1=0.268; average value of body length X2=11.37cm, standard deviation SV2=2.15, discrete coefficient CV2=0.189; mean value of reproductive band diameter X3=5.4mm, standard deviation SV3=0.82, discrete The coefficient CV3 = 0.15; the average value of the vitality X4 = 42.9 s, the standard deviation SV4 = 6.21, and the dispersion coefficient CV4 = 0.148. At the same time, it satisfies X1≥5.5g, CV1≤0.28, X2≥11.0cm, CV2≤0.2, X3≥5.0mm, CV3≤0.16, X4≥42s and CV4≤0.15, so it is judged that the wide dragon of Guangdilong breeding base in Luchuan County The seedling population is a qualified population of the genus Longjing seedlings.

Claims (10)

1. The method for evaluating individual quality of the radix chinensis seedlings comprises the following steps: observing the individual integrity, body color and reproductive zonal band of the genus, and measuring the individual weight, body length, reproductive ring diameter and Vitality; if the Guanglong dragon is intact, the back is tan to purple gray, the abdomen is light yellow-brown, the reproductive ring is not obvious, and at the same time, the individual weight of Guangdilong is ≥4.0g, the body length is ≥9.0cm, and the reproductive ring diameter is ≥4.5. When mm and vigor are ≥35.0s, it is judged that the Guanglong individual is a qualified Guangdi Longyan seedling.
2. The method according to claim 1, wherein the body length of the genus of the genus is the length of the reproductive ring to the tail.
3. The method according to claim 1, wherein the measuring the body length of the Phyllostachys pubescens is specifically to place the cockroach in a volume fraction of 50% alcohol, and to anesthetize it. Until the body is in a relaxed state, measure the length of the reproductive ring to the tail.
4. The method according to claim 1, wherein the measuring the diameter of the reproductive ring of the genus is specifically measuring the diameter of the reproductive ring in the quiet state of the terrapin.
5. The method according to claim 1, wherein the measuring the activity of the scorpion dragon is specifically to place the phoenix dragon in a volume fraction of 50% alcohol, and recording the anesthesia to The time it takes for the body to be in a relaxed state.
6. A method for evaluating the quality of the seedlings of the radix chinensis seedlings is characterized in that it comprises the following steps: randomly collecting a plurality of genus genus, measuring the individual weight, body length, reproductive band diameter and vitality of the genus, and calculating the collection The average value of the individual weight of the broad-eared dragon X1 and the standard deviation SV1, the average value of the body length of the wide-body dragon X2 and the standard deviation SV2, the average value of the diameter of the individual reproductive belt ring X3 and the standard deviation SV3, Guangdilong The mean value of individual vitality X4 and standard deviation SV4, according to the average weight and standard deviation of the individual weight, body length, reproductive ring diameter and vigor of the genus, the individual weight, body length and reproductive ring diameter of the genus And the discrete coefficients of vitality CV1, CV2, CV3 and CV4; if simultaneously satisfy X1≥5.5g, CV1≤0.28, X2≥11.0cm, CV2≤0.2, X3≥5.0mm, CV3≤0.16,X4≥42s and CV4≤0.15 Then, it is judged that the population of the long-range seedlings is a qualified population of Longjing seedlings.
7. The method according to claim 6, wherein the body length of the genus of the genus is the length of the reproductive ring to the tail.
8. The method for evaluating the quality of the seedlings of the genus Amaranthus chinensis according to claim 6, wherein the measuring body length of the Phyllostachys pubescens is specifically to place the cockroach in a volume fraction of 50% alcohol, and wait for the Anesthesia until the body is in a relaxed state, measuring the length of the reproductive ring to the tail.
9. The method according to claim 6, wherein the measuring the diameter of the reproductive ring of the genus is specifically measuring the diameter of the reproductive ring in the quiet state of the terrapin.
10. The method for evaluating the quality of the seedlings of the genus Amaranthus chinensis according to claim 6, wherein the measuring the activity of the genus sylvestris is specifically to place the phoenix in a volume fraction of 50% alcohol, and record the anesthesia. The time until the body is in a relaxed state.

Images