CN111264124A

CN111264124A - Method for efficiently promoting germination of seeds of premna hirta

Info

Publication number: CN111264124A
Application number: CN202010254178.4A
Authority: CN
Inventors: 张明生; 幸菲菲; 刘剑东; 李扬; 阳娇; 王艳秋
Original assignee: Guizhou University
Current assignee: Guizhou University
Priority date: 2020-04-02
Filing date: 2020-04-02
Publication date: 2020-06-12

Abstract

The invention discloses a method for efficiently promoting germination of bromhidrosis firewood seeds. The method comprises the following steps: soaking mature wood fruit with bromhidrosis in clear water, removing peel, cleaning, and selecting clean seeds; soaking the seeds for later use in sulfuric acid solution for 3-8min, washing with clear water, and then sucking dry water on the surface of the seeds with filter paper for later use; soaking the seeds for later use in gibberellin solution, taking out, washing with clear water, and absorbing water on the surfaces of the seeds with filter paper for later use; sterilizing the seeds for later use by using 75% of alcohol, 0.1% of mercury bichloride and sterile water for washing to obtain sterilized seeds for later use; inoculating the disinfected seeds into an MS culture medium for germination culture. The method has the advantages of effectively solving the problem of difficult germination of the corymbose odoriphaga seeds caused by physical dormancy, improving the germination rate of the corymbose odoriphaga seeds and shortening the germination time, thereby providing guarantee for a large-scale production technology, along with simple operation and low cost.

Description

Method for efficiently promoting germination of seeds of premna hirta

Technical Field

The invention relates to a method for germinating hircismus bunge seeds, in particular to a method for efficiently promoting the germination of the hircismus bunge seeds.

Background

Bromhidrosis firewood (Premnamberlla Pamp), also known as immortal Premna microphylla, belongs to erect or shrub to small arbor of Premna microphylla of Verbenaceae, has the effects of clearing heat and removing toxicity, and relieving swelling and stopping bleeding, and has very wide development and utilization values. The bromhidrosis firewood is seriously felled under the wild condition, lacks reasonable development, management and utilization, is scattered, and the number of adult plants is continuously reduced, so that the artificial planting is necessary for realizing the industrialized development of the bromhidrosis firewood resource.

At present, according to the literature, the natural propagation and germination of the seeds of the corylus odorifera in the natural state are difficult, and experiments prove that the seed coat is hard, so that the physical dormancy phenomenon of the seeds exists, which is a main factor for limiting the germination of the corylus odorifera. Therefore, the method can be used for rapidly and efficiently germinating the hircismus seeds by removing the germination obstacle of the hircismus seeds, improving the germination rate, and is a key link for obtaining high-quality seed seedlings for breeding and realizing large-scale artificial planting.

Disclosure of Invention

The invention aims to provide a method for efficiently promoting germination of the seeds of the premna hirta. The method has the characteristics of effectively solving the problem of difficult germination of the corymbose odoriphaga seeds caused by physical dormancy, improving the germination rate of the corymbose odoriphaga seeds and shortening the germination time, thereby providing guarantee for a large-scale production technology, along with simple operation and low cost.

The technical scheme of the invention is as follows: a method for efficiently promoting the germination of the seeds of the premna hirta comprises the following steps:

the method comprises the following steps: soaking mature fruits of the premna hirta in clear water for 12-24h, removing peel, cleaning, and selecting clean seeds for later use;

step two: soaking the standby seeds in the first step in a sulfuric acid solution for 3-8min, washing the seeds with clear water, and then sucking water on the surfaces of the seeds by using filter paper for standby;

step three: placing the seeds for later use in the step two into a gibberellin solution, soaking for 12-36h, taking out, washing with clear water, and then sucking water on the surfaces of the seeds by using filter paper for later use;

step four: sterilizing the spare seeds obtained in the third step by using 75% alcohol for 0.5-2min, 0.1% mercury bichloride for 3-10min and sterile water for several times to obtain sterilized seeds for later use;

step five: inoculating the seeds sterilized in the step four into an MS culture medium for germination culture, wherein the culture time is 25-35 d.

In the method for efficiently promoting the germination of the bromhidrosis firewood seeds, in the second step, the mass concentration of the sulfuric acid solution is 98%.

In the aforementioned method for efficiently promoting the germination of the seeds of the corylus bromhidrosis, in the third step, the concentration of the gibberellin solution is 100-300 mg/L.

In the fourth step, the seeds for standby in the third step are firstly soaked in 75% alcohol for 0.5-2min, then washed with sterile water for 1-3 times, then soaked in 0.1% mercuric chloride for 3-10min, washed with sterile water for 2-4 times, and finally the seeds after disinfection treatment are obtained by sucking or draining water through filter paper for standby.

In the fifth step, the content of the MS culture medium is 1.5-2.5 mg/L6-BA and 0-1mg/L GA₃25-35g/L of sucrose, 5-8g/L of agar and pH of 5.8-6.0.

In the aforementioned method for efficiently promoting the germination of the corylus chinensis seeds, in the fifth step, the germination culture condition is illumination intensity of 2000-3000lx, the illumination time is 12h/d, and the culture temperature is 25 +/-2 ℃.

Compared with the prior art, the invention has the following beneficial effects:

concentrated sulfuric acid with the mass concentration of 98 percent and plant hormone gibberellin GA are utilized₃The method for treating the hircismus bunge seeds by combining soaking can ensure that the hircismus bunge seeds germinate in a short time and improve the germination rate, and effectively solves the problem that the hircismus bunge seeds are difficult to germinate due to the fact that seed coats are in physical dormancy.

The inventors carried out a number of experiments, and the following were part of the experimental studies:

FIG. 1 is a diagram of seed germination cultivation in example 1 "soaking seeds in 98% concentrated sulfuric acid solution for 6 min";

the inventor's "soak the seeds in the concentrated sulfuric acid solution with the mass concentration of 98% for 3 min", the other conditions are the same as the example 1, and a seed germination culture diagram is obtained, which is shown in fig. 2;

the inventor soaks the seeds in a concentrated sulfuric acid solution with the mass concentration of 98% for 8min, and obtains a seed germination culture diagram which is shown in a figure 3 under the same other conditions as the example 1;

the inventor omits the prior treatment of the seeds in the embodiment 1, and directly takes the mature fruits of the premna hirta in 6-7 months for disinfection treatment and germination culture to obtain a seed germination culture diagram, which is shown in figure 4;

as can be seen from comparative analysis of fig. 1, 2 and 3, in example 1, "soaking seeds in a concentrated sulfuric acid solution with a mass concentration of 98% for 6 min" is the optimal parameter selection, and the germination rate is high; as can be seen from the comparative analysis of fig. 1, 2, 3 and 4, the preliminary treatment of the seeds in embodiment 1 of the present invention "soaking in clear water for 24 hours, removing the peel with tweezers, and selecting clean seeds after washing; soaking the seeds in a concentrated sulfuric acid solution with the mass concentration of 98% for 6min, stirring the seeds during the soaking, washing the seeds clean with clear water, and then absorbing water on the surfaces of the seeds with filter paper; and then the seeds are placed in a gibberellin solution of 200mg/L to be soaked for 24 hours and then taken out, and the seeds are washed clean by clear water and then are dried by filter paper for standby application, so that the method has the advantages of improving the germination rate of the seeds of the corymbifera odorata and shortening the germination time.

In conclusion, the method has the advantages of effectively solving the problem of difficult germination of the corymbose odoriphaga seeds caused by physical dormancy, improving the germination rate of the corymbose odoriphaga seeds and shortening the germination time, thereby providing guarantee for a large-scale production technology, along with simple operation and low cost.

Drawings

FIG. 1 is a diagram showing seed germination under the condition of "immersing seeds in a concentrated sulfuric acid solution having a mass concentration of 98% for 6 min" according to the present invention (a diagram showing seed germination in example 1);

FIG. 2 is a diagram of seed germination under the condition of "soaking seeds in a concentrated sulfuric acid solution with a mass concentration of 98% for 3 min" according to the present invention;

FIG. 3 is a diagram of seed germination under the condition of "soaking seeds in a concentrated sulfuric acid solution with a mass concentration of 98% for 8 min" according to the present invention;

FIG. 4 is a seed germination culture diagram obtained by directly sterilizing and germinating mature fruits of Premna microphylla in 6-7 months without the pretreatment of the seeds.

Detailed Description

The invention is further illustrated by the following figures and examples, which are not to be construed as limiting the invention.

Example 1. A method for efficiently promoting germination of bromhidrosis firewood seeds is characterized in that plant material bromhidrosis firewood (Premnauberula Pamp.) in the embodiment is collected from the autonomous county of Silao Miao nationality in Guizhou province and is identified by the New professor of Xiongyuan of Guizhou university.

Example 1

Pre-treatment of seeds: soaking mature fruits of Bupleurum falcatum in clear water for 24h for 6-7 months, removing peel with tweezers, cleaning, and selecting clean seeds; soaking the seeds in a concentrated sulfuric acid solution with the mass concentration of 98% for 6min, stirring the seeds during the soaking, washing the seeds clean with clear water, and then absorbing water on the surfaces of the seeds with filter paper; then placing the seeds in 200mg/L gibberellin solution to be soaked for 24 hours, taking out the seeds, washing the seeds clean with clear water, and then sucking the water on the surfaces of the seeds with filter paper for later use;

and (3) disinfection treatment: placing the seeds for standby in a sterilized beaker, sterilizing the surface of the beaker with alcohol, placing the beaker in an aseptic operation table, sequentially washing with 75% alcohol for 1min and sterile water for 2 times, soaking for 8min with 0.1% mercuric chloride for 1min each time, washing with sterile water for 3 times, and draining water for standby after cleaning the seeds for 1min each time;

germination and culture: placing the sterilized seeds on sterile filter paper in a sterile operation table, sucking water, inoculating in MS culture medium, and performing germination culture, wherein the MS culture medium is respectively added with 2.0 mg/L6-BA and 0.5mg/L GA₃30g/L of sucrose and 7g/L of agar; the culture conditions are pH5.8-6.0, illumination intensity 2000-The culture time is 12h/d, the culture temperature is 25 +/-2 ℃, and the culture time is 25-35 d.

Claims

1. A method for efficiently promoting the germination of the seeds of the premna hirta is characterized by comprising the following steps: the method comprises the following steps:

2. The method for efficiently promoting the germination of the seeds of the premna hirta as claimed in claim 1, wherein the method comprises the following steps: in the second step, the mass concentration of the sulfuric acid solution is 98%.

3. The method for efficiently promoting the germination of the seeds of the premna hirta as claimed in claim 1, wherein the method comprises the following steps: in the third step, the concentration of the gibberellin solution is 100-300 mg/L.

4. The method for efficiently promoting the germination of the seeds of the premna hirta as claimed in claim 1, wherein the method comprises the following steps: in the fourth step, the seeds for standby in the third step are soaked in 75% alcohol for 0.5-2min, washed with sterile water for 1-3 times, then soaked in 0.1% mercuric chloride for 3-10min, washed with sterile water for 2-4 times, and then the seeds after disinfection treatment are obtained by absorbing or draining water with filter paper.

5. The method for efficiently promoting the germination of the seeds of the premna hirta as claimed in claim 1, wherein the method comprises the following steps: in the fifth step, the content of MS culture medium is 1.5-2.5 mg/L6-BA and 0-1mg/L GA₃25-35g/L of sucrose, 5-8g/L of agar and pH5.8-6.0.

6. The method for efficiently promoting the germination of the seeds of the premna hirta as claimed in claim 1, wherein the method comprises the following steps: in the fifth step, the germination culture condition is illumination intensity of 2000-3000lx, the illumination time is 12h/d, and the culture temperature is 25 +/-2 ℃.