阅读说明：本技术 一种提高甘薯脱毒组培苗移栽成活率的方法 (Method for improving transplanting survival rate of sweet potato virus-free tissue culture seedlings ) 是由 陈选阳 陈杏婷 陈国太 尼加提·努尔拉 黄小芳 毕楚韵 杨志坚 林世强 于 2019-10-18 设计创作，主要内容包括：本发明公开了一种提高甘薯脱毒组培苗移栽成活率的方法,包括配制培育壮苗培养基配方、水培炼苗与适时移栽、设置移栽环境、适时移栽和移栽后管理。本发明在组培苗阶段,培养基中添加了多效唑使苗生产更健壮；在炼苗阶段采用水培方式,使其环境处于组培与室外生长的中间过渡环境；在移栽后,控制适宜的环境湿度和温度；这样可以培育组培苗壮苗,促进组培苗长根,提高组培苗移栽成活率。(The invention discloses a method for improving the transplanting survival rate of sweet potato virus-free tissue culture seedlings, which comprises the steps of preparing a culture medium formula for cultivating strong seedlings, hardening seedlings by water culture and transplanting in good time, setting a transplanting environment, transplanting in good time and managing after transplanting. In the tissue culture seedling stage, paclobutrazol is added into a culture medium, so that the seedlings are more robust; in the hardening-seedling stage, a water culture mode is adopted, so that the environment is in a transition environment between tissue culture and outdoor growth; after transplanting, controlling proper environmental humidity and temperature; therefore, strong seedlings of the tissue culture seedlings can be cultured, the tissue culture seedlings are promoted to grow roots, and the transplanting survival rate of the tissue culture seedlings is improved.)

1. A method for improving the transplanting survival rate of sweet potato virus-free tissue culture seedlings is characterized by comprising the following steps:

(1) preparing a culture medium formula for strengthening sweet potato virus-free tissue culture seedlings and carrying out tissue culture;

(2) water planting and hardening seedlings;

(3) setting a transplanting environment, and keeping higher environmental humidity and loose soil;

(4) transplanting;

(5) and (5) managing after transplanting.

2. The method for improving the transplanting survival rate of the sweet potato virus-free tissue culture seedlings according to claim 1, wherein the step (1) comprises the following steps:

1) the formula for preparing the culture medium for strengthening the sweet potato virus-free tissue culture seedlings comprises the following steps: MS + 0.6 mg/L paclobutrazol + 0.15 mg/L6-benzylamino adenine + 0.15 mg/L naphthylacetic acid, pH5.8;

2) culturing sweet potato tissue culture seedlings: selecting a normally growing detoxified tissue culture seedling, shearing stem segments with 2 leaves, and inserting the stem segments into a culture medium of a tissue culture bottle; then, the cells were cultured in an environment of 1600Lx light, 16/8 hours light/dark time, and 26 ℃.

3. The method for improving the transplanting survival rate of the sweet potato virus-free tissue culture seedlings according to claim 1, wherein the step (2) specifically comprises the following steps:

1) making a seedling hardening water culture device: selecting a water culture tank with the length of 60cm, the width of 20cm and the height of 20cm, keeping pure water with the depth of 10cm in the tank, preparing a foam plate with the thickness of 0.5cm, wherein the length and the width are slightly smaller than the middle section of the water culture tank, and drilling two rows of planting holes in the middle of the foam plate, wherein the distance between the two rows of planting holes is 10cm, and the distance between the holes is 5 cm; covering a transparent preservative film above the water culture tank;

2) water culture tissue culture seedling: selecting the tissue culture seedlings which grow in the tissue culture bottle for 30 days in the step (1), have 6-8 leaves and have the plant height of 10cm, cutting off root systems, inserting the tissue culture seedlings into the transplanting holes of the foam board, exposing 2cm stem sections at the lower ends of the tissue culture seedlings, and soaking the tissue culture seedlings in pure water in a water culture tank for water culture seedling hardening.

4. The method for improving the transplanting survival rate of the sweet potato virus-free tissue culture seedlings according to claim 3, wherein during the water culture hardening period, the water surface height of a water culture tank is kept at 10cm, the environmental temperature is 25-28 ℃, and the hardening time is 10 days.

5. The method for improving the transplanting survival rate of the sweet potato virus-free tissue culture seedlings according to claim 1, wherein the step (3) comprises the following steps:

1) setting a tissue culture seedling transplanting environment: in the field, a plastic shed is built by using steel pipes or bamboo chips, a plastic film and a sunshade net are covered on the plastic shed, the humidity in the shed is kept between 90 and 95 percent, and the temperature is kept between 25 and 30 ℃;

2) preparing loose soil: the loose soil is prepared by river sand and soil in a ratio of 1: 1.

6. The method for improving the transplanting survival rate of the sweet potato virus-free tissue culture seedlings according to claim 1, wherein the step (4) comprises the following steps: and (3) when the tissue culture seedlings after water culture hardening in the step (2) are transplanted, bending the two blades at the base parts of the tissue culture seedlings downwards, burying the two blades and the root systems of the tissue culture seedlings into soil and compacting the blades, and then pouring root fixing water, wherein the planting depth of the tissue culture seedlings is 3 cm.

7. The method for improving the survival rate of transplanted detoxified tissue culture seedlings according to claim 1, wherein the step (5) comprises the following steps: after the tissue culture seedlings are transplanted, a sunshade net is covered on the shed, water is sprayed for 7 days continuously, humidity is kept, and the tissue culture seedlings are promoted to grow roots and survive.

Technical Field

The invention belongs to the field of crop cultivation and biotechnology, and particularly relates to a method for improving the transplanting survival rate of sweet potato virus-free tissue culture seedlings.

Background

Sweet potatoes are important crops of grains, feeds and industrial raw materials in China. The cultivation area and the total yield all account for the first place in the world. However, sweet potato production faces the threat of various virus diseases, and particularly Sweet Potato Virus Diseases (SPVD) which can cause outages in recent years are introduced into China and have the possibility of gradual diffusion. The best method for preventing virus diseases in production is to popularize and apply the virus-free seedlings. However, the virus-free seedlings used in production account for only a small proportion at present. Therefore, the detoxified seedlings need to be widely popularized and applied to ensure the safety of the sweet potato industry.

At present, before the tissue culture virus-free seedlings are transplanted, strong seedlings are not cultured specially, hardening seedlings before transplanting are carried out, the culture medium is washed off after the bottle cap of the tissue culture is opened to harden the seedlings for several days, and the tissue culture seedlings are transplanted into the matrix with roots. The roots in the culture medium generally have no root hairs, and are difficult to adapt to the new environment of matrix soil, so that the survival rate of tissue culture seedling transplantation is influenced.

Disclosure of Invention

The technical problem to be solved by the invention is to provide a method for improving the transplanting survival rate of the detoxified tissue culture seedlings aiming at the defects in the prior art, so as to improve the propagation speed of the detoxified seedlings of the sweet potatoes, provide healthy and healthy fine-breed seedlings for production and ensure the production safety.

A method for improving the transplanting survival rate of sweet potato virus-free tissue culture seedlings comprises the following steps:

(1) preparing a culture medium formula for strengthening the sweet potato virus-free seedlings and carrying out tissue culture;

(2) hardening seedlings by water culture and transplanting in due time;

(3) setting a transplanting environment, and keeping higher environmental humidity and loose soil;

(4) transplanting;

(5) and (5) managing after transplanting.

The steps specifically include the following steps:

1) preparing a culture medium formula for the virus-free seedling and strong seedling of the sweet potato, wherein MS, 0.6 mg/L paclobutrazol, 0.15 mg/L6-benzylamino adenine, 0.15 mg/L naphthylacetic acid and pH5.8 are subjected to autoclaving for later use;

2) selecting a normally growing detoxified tissue culture seedling, shearing stem segments with 2 leaves, and inserting the stem segments into a culture medium of a tissue culture bottle; the tissue culture seedlings are subjected to strong seedling culture in an environment with the illumination of 1600Lx, the illumination/dark time of 16/8 hours and the temperature of 26 ℃.

3) Making a seedling hardening water culture device: selecting a water culture tank with the length of 60cm, the width of 20cm and the height of 20cm, keeping pure water with the depth of 10cm in the tank, preparing a foam plate with the thickness of 0.5cm, slightly reducing the length and the width to the middle section of the water culture tank, drilling two rows of planting holes in the middle of the foam plate, wherein the distance between the two rows of planting holes is 10cm, and the distance between the holes is 5 cm. And a transparent preservative film is covered above the water culture tank.

4) Water culture seedling hardening: selecting sweet potato tissue culture seedling which grows in a tissue culture bottle for about 30 days and has 6-8 leaves and plant height of about 10cm, cutting off root system, inserting the tissue culture seedling into a foam board to expose 2cm stem segment at the lower end of the tissue culture seedling, and soaking in pure water in a water culture tank. During the hardening period, the water surface height of the water culture tank is kept at 10cm, the environmental temperature is kept at 25-28 ℃, and the hardening time is 10 days.

5) Setting a sweet potato tissue culture seedling transplanting environment: in the field, a plastic shed is built by using steel pipes or bamboo chips, and the shed can be covered with a plastic film and a sunshade net, so that the humidity in the shed is kept between 90 and 95 percent, and the temperature is kept between 25 and 30 ℃;

6) preparing loose soil: the loose soil is prepared from river sand and vegetable garden soil in a ratio of 1: 1.

7) Transplanting the tissue culture seedling, bending two blades at the base part of the sweet potato tissue culture seedling downwards, burying the two blades and the root system of the tissue culture seedling into soil, slightly compacting, wherein the planting depth of the tissue culture seedling is 3cm, and immediately watering for rooting after transplanting.

8) After the tissue culture seedlings are transplanted, a sunshade net is covered on the shed to prevent strong light irradiation; and water is required to be sprayed for 7 days after transplanting, the humidity is kept, and the growth and the survival of the tissue culture seedlings are promoted.

The invention has the following remarkable advantages:

1. after strong seedling culture, the tissue culture seedling has stronger stress resistance than a control.

2. The water culture hardening seedling setting is simple and the operation is easy.

3. After hardening-seedling treatment, the transplanting survival rate of the detoxified tissue culture seedlings of the sweet potatoes can reach 100 percent.

Detailed Description