阅读说明：本技术 一种提高玉米单倍体加倍效率的栽培方法 (Cultivation method for improving doubling efficiency of corn haploid ) 是由 刘文国 孟令聪 何欢 周德龙 张建新 郑淑波 高嵩 岳尧海 路明 周旭东 王敏 于 2021-08-24 设计创作，主要内容包括：本发明公开了一种提高玉米单倍体加倍效率的栽培方法,属于作物栽培技术领域。本发明公开的一种提高玉米单倍体加倍效率的栽培方法,采用专用营养土,加入营养液,将玉米单倍体幼苗移入牛皮纸袋后,直接进入大棚内进行移栽和炼苗缓苗,保持棚内温度、湿度,炼苗,喷施专用营养液和消毒液,保持牛皮纸袋里土壤湿润,田间精心管理。本发明方法省略了室外炼苗缓苗过程,减少了大棚内移栽时对幼苗的机械损伤,可以使单倍体幼苗成活率达约80-90％,从而可以提高玉米单倍体加倍率约25-34％,降低了育种材料和人工成本,减少了环境污染,提高了单倍体育种效率,操作简便,可大规模推广应用。(The invention discloses a cultivation method for improving doubling efficiency of corn haploid, and belongs to the technical field of crop cultivation. The invention discloses a cultivation method for improving doubling efficiency of corn haploid, which adopts special nutrient soil, adds nutrient solution, directly enters a greenhouse for transplanting, hardening and seedling recovering after transplanting corn haploid seedlings into a kraft paper bag, keeps temperature and humidity in the greenhouse, hardens seedlings, sprays special nutrient solution and disinfectant, keeps soil in the kraft paper bag moist, and carefully manages in the field. The method omits the outdoor seedling exercising and reviving process, reduces the mechanical damage to the seedlings during transplanting in a greenhouse, and can ensure that the survival rate of the haploid seedlings reaches about 80-90 percent, thereby improving the corn haploid multiplying power by about 25-34 percent, reducing the breeding material and labor cost, reducing the environmental pollution, improving the haploid breeding efficiency, having simple and convenient operation and being suitable for large-scale popularization and application.)

1. A cultivation method for improving doubling efficiency of corn haploid comprises building a plastic greenhouse, preparing special nutrient soil, preparing nutrient solution and disinfectant, preparing seedling, transplanting seedlings in the greenhouse and managing the field; the method is characterized in that special nutrient soil is adopted, nutrient solution is added, corn haploid seedlings are transplanted into a kraft paper bag, then the corn haploid seedlings directly enter a greenhouse to be transplanted and hardened and slow seedlings, the temperature and the humidity in the greenhouse are kept at 28-32 ℃ and 60-75%, the seedlings are hardened for 10-15 days, the special nutrient solution and disinfectant are sprayed every 2-3 days, the soil in the kraft paper bag is kept moist, and field management is meticulous.

2. The cultivation method for improving the doubling efficiency of the corn haploid as claimed in claim 1, which is characterized by comprising the following specific steps:

(1) building a plastic greenhouse: the appearance of the greenhouse is built in an arched shape, the height in the greenhouse is 1.8-2.0m, the arched part of the greenhouse is covered by a special vegetable greenhouse film, and the periphery of the greenhouse is built by a special vegetable greenhouse insect-proof net to keep ventilation and insect prevention in the greenhouse;

(2) preparing special nutrient soil: mixing danish peaty peat soil, perlite and turfy soil according to the proportion of 5: 1: 4, obtaining special nutrient soil for later use;

(3) preparing nutrient solution and disinfectant: mixing the nutrient fertilizer according to the proportion of 1: 500, and mixing the rooting powder according to the proportion of 1: 200, diluting, and mixing the diluted nutrient fertilizer and the diluted rooting powder in equal volume to prepare a special mixed nutrient solution for later use; mixing the sterilizing agent according to the proportion of 1: 1000, diluting and uniformly stirring to prepare a disinfectant for later use;

(4) seedling preparation: spraying the special nutrient soil prepared in the step (2) by adopting the nutrient solution prepared in the step (3), wherein the special nutrient soil keeps a semi-dry and semi-wet state, and then filling the semi-dry and semi-wet special nutrient soil into a kraft paper bag, wherein the soil filling amount is two thirds of the capacity of the paper bag;

(5) transplanting seedlings: taking out the corn haploid seedlings in the laboratory culture bottles, and carefully transferring the corn haploid seedlings into the kraft paper bags in the step (4);

(6) transplanting in a greenhouse: manually digging a ditch with the depth of 15-20cm, placing a kraft paper bag containing corn haploid seedlings into the ditch, manually filling and slightly pressing a small amount of fine soil around the paper bag; sprinkling irrigation is carried out in time after the transplanting is finished;

(7) field management: keeping the temperature in the greenhouse at 28-32 ℃ and the humidity at 60-75%, hardening seedlings for 10-15 days, spraying special nutrient solution and disinfectant once every 2-3 days on the special nutrient soil in the kraft bags, spraying 40-50ml of special nutrient solution and 10-15ml of disinfectant once on each kraft bag, keeping the soil in the kraft bags moist, and fertilizing when the seedlings grow to 6-7 leaves; the field management work is carried out by a specially-assigned person, so that sufficient moisture in the shed is ensured, and weeds are avoided.

Technical Field

The invention relates to the technical field of crop cultivation, in particular to a cultivation method for improving doubling efficiency of corn haploid.

Background

In recent years, haploid breeding technology of corn based on biological induction has gradually become a key technology of corn breeding. The Doubled Haploid (DH) technique is the fastest, simplest, economical, practical and direct method for breeding maize inbred lines. The basic process is to obtain homozygous diploid plant by artificially or naturally doubling the biologically induced haploid plant, and then to screen the superior line from the homozygous diploid plant. The method is mainly characterized in that the line selection speed is high, and a stable pure line can be obtained only by 2 generations (more than 7 generations are needed for traditional inbred line breeding).

With the breeding of novel induction lines (Nongda high-inducing No. 1, Jigao inducing No. 3 and Nongda high-inducing No. 5), the induction efficiency and the identification efficiency are continuously improved, and the doubling technology of haploids is the bottleneck of the current large-scale application. Haploid doubling methods are numerous, physical and chemical factors are applied to treatment, and the method can be successfully applied, and is mainly divided into natural doubling and chemical artificial doubling, the natural doubling frequency is usually low, and the natural doubling rate of most materials is lower than 5%. The chemical doubling is more effective, and the doubling rate can be obviously improved to 15-20%. Therefore, the application of the chemical doubling method is an effective method for improving the haploid breeding efficiency. At present, in the chemical doubling method, the doubling rate of the young embryo doubling method can reach 20-30%, and the method is a high-efficiency doubling method, but the outdoor seedling exercising and seedling reviving and transplanting processes are complicated, and the doubling effect of the young embryo can be directly influenced.

At present, the conventional cultivation procedure of the doubled haploid embryo seedlings comprises the steps of building a shading net, preparing special nutrient soil, preparing nutrient solution and disinfectant, preparing seedling, transplanting seedlings, exercising and reviving seedlings outdoors, transplanting in the field and managing in the field. Wherein, the outdoor hardening-seedling and transplanting process is very important and needs to be carefully cultured and managed. The conventional seedling hardening and reviving and transplanting processes are carried out by adopting a sunshade net, and the sprayed solution has single nutrition, insufficient illumination time, lower growth temperature, mechanical damage to seedlings and the like, so that the survival rate of the seedlings is greatly reduced. The doubling efficiency and scale are influenced, and the breeding efficiency is reduced.

Therefore, the problem to be solved by the technical personnel in the field is to provide a cultivation method for improving the doubling efficiency of the corn haploid.

Disclosure of Invention

In view of the above, the cultivation method for improving the doubling efficiency of the corn haploid is provided, is simple and convenient to operate, is environment-friendly, is low in cost, greatly improves the doubling efficiency, and has important significance for large-scale breeding of the corn haploid.

In order to achieve the purpose, the invention adopts the following technical scheme:

a cultivation method for improving doubling efficiency of corn haploid comprises building a plastic greenhouse, preparing special nutrient soil, preparing nutrient solution and disinfectant, preparing seedling, transplanting seedlings in the greenhouse and managing the field; the method comprises the steps of adopting special nutrient soil, adding nutrient solution, directly entering a greenhouse for transplanting, hardening and seedling recovering after transferring corn haploid seedlings into a kraft paper bag, keeping the temperature of 28-32 ℃ and the humidity of 60-75% in the greenhouse, hardening seedlings for 10-15 days, spraying the special nutrient solution and disinfectant once every 2-3 days, keeping the soil in the kraft paper bag moist, and carefully managing in the field.

Further, the cultivation method for improving the doubling efficiency of the corn haploid comprises the following specific steps:

(1) building a plastic greenhouse: the appearance of the greenhouse is built in an arched manner, the height in the greenhouse is 1.8-2.0m, the arched part of the greenhouse is covered by a special vegetable greenhouse film which has high light transmittance, heat preservation and wind resistance, and the periphery of the greenhouse is built in a surrounding manner by a special vegetable greenhouse insect-proof net with high density, so that ventilation and insect prevention in the greenhouse are kept;

(2) preparing special nutrient soil: mixing danish peaty peat soil, perlite and turfy soil according to the proportion of 5: 1: 4, obtaining special nutrient soil for later use;

(3) preparing nutrient solution and disinfectant: mixing the nutrient fertilizer according to the proportion of 1: 500, and mixing the rooting powder according to the proportion of 1: 200, diluting, and mixing the diluted nutrient fertilizer and the diluted rooting powder in equal volume to prepare a special mixed nutrient solution for later use; mixing the sterilizing agent according to the proportion of 1: 1000, diluting and uniformly stirring to prepare a disinfectant for later use;

(4) seedling preparation: spraying the special nutrient soil prepared in the step (2) by adopting the nutrient solution prepared in the step (3), wherein the special nutrient soil keeps a semi-dry and semi-wet state, and then filling the semi-dry and semi-wet special nutrient soil into a kraft paper bag, wherein the soil filling amount is two thirds of the capacity of the paper bag;

(5) transplanting seedlings: taking out the corn haploid seedlings in the laboratory culture bottles, and carefully transferring the corn haploid seedlings into the kraft paper bags in the step (4);

(6) transplanting in a greenhouse: manually digging a ditch with the depth of 15-20cm, placing a kraft paper bag containing corn haploid seedlings into the ditch, manually filling and slightly pressing a small amount of fine soil around the paper bag; sprinkling irrigation is carried out in time after the transplanting is finished;

(7) field management: keeping the temperature in the greenhouse at 28-32 ℃ and the humidity at 60-75%, hardening seedlings for 10-15 days, spraying special nutrient solution and disinfectant once every 2-3 days on the special nutrient soil in the kraft bags, spraying 40-50ml of special nutrient solution and 10-15ml of disinfectant once on each kraft bag, keeping the soil in the kraft bags moist, and fertilizing when the seedlings grow to 6-7 leaves; the field management work is carried out by a specially-assigned person, so that sufficient moisture in the shed is ensured, and weeds are avoided.

Compared with the conventional culture technology of the seedling after the doubling of the haploid embryo, the invention provides the culture method for improving the doubling efficiency of the corn haploid. The technology has the outstanding innovation that corn haploid seedlings cultured in a laboratory are transplanted into a kraft paper bag and directly enter a greenhouse for transplanting, hardening and seedling recovering, the temperature and the humidity in the greenhouse are kept at 28-32 ℃ and 60-75%, the seedling is hardened for 10-15 days, special nutrient solution and a small amount of disinfectant are sprayed every 2-3 days, the soil is kept moist, the haploid seedlings can grow healthily, the outdoor seedling hardening and recovering process is omitted, the damage rate of the seedlings is reduced, the survival rate of the haploid seedlings can reach about 80-90%, the corn haploid multiplying power can be increased by about 25-34%, the breeding material and labor cost are reduced, the environmental pollution is reduced, the haploid breeding efficiency is improved, the operation is simple and convenient, and the technology can be popularized and applied in a large scale. The method can be repeatedly implemented, has strong reproducibility, and greatly improves the survival rate of the seedlings.

According to the technical scheme, compared with the prior art, the cultivation method for improving the doubling efficiency of the corn haploid provided by the invention has the following beneficial effects:

(1) the corn haploid seedlings are cultured by adopting special nutrient soil, nutrient solution and kraft paper bags, so that the seedling damage can be reduced, the seedlings can be revived more easily, and the survival rate of the seedlings is improved.

(2) The kraft paper bag adopted for seedling cultivation is cheaper than the seedling cultivation pot and the seedling revival frame, so that a lot of material cost is saved, and the operation is simple and convenient.

(3) After the corn haploid seedlings are transplanted into the kraft paper bags, the corn haploid seedlings directly enter a greenhouse for transplanting, the seedling hardening and seedling reviving process of adopting seedling pots and seedling racks outdoors in the conventional seedling reviving cultivation method is omitted, and much labor cost is saved.

(4) In the field seedling transplanting process in the conventional seedling revival cultivation method, the corn haploid seedlings need to be poured out of the seedling pot or pulled out by hands for transplanting, the mechanical damage to the seedlings is large, and the mechanical damage is not beneficial to revival; the invention transplants the seedlings contained in the kraft paper bags by directly placing the paper bags containing the seedlings into the ditch, which hardly damages the seedlings and ensures that the seedlings can survive more easily.

(5) The greenhouse adopts a special vegetable greenhouse film with high light transmittance, heat preservation and wind prevention to buckle the film, so that the light transmission in the greenhouse is good, the higher temperature is kept, and the high-density special vegetable greenhouse insect-proof net is adopted around the greenhouse, so that the ventilation in the greenhouse is good, insect damage is prevented, the temperature and humidity in the greenhouse are adjusted, the growth of seedlings is promoted, and the survival of the seedlings is facilitated.

(6) After the corn haploid seedlings are moved into a shed, direct sunlight irradiation is adopted to revive the seedlings, the temperature in the shed is kept at 25-32 ℃, the humidity is kept at 60-75%, the seedlings are trained for 10-15 days, and special nutrient solution and a small amount of disinfectant are sprayed every 2-3 days to keep the soil moist, so that the haploid seedlings can grow healthily, the damage rate of the seedlings is reduced, and the survival rate and the doubling rate of the seedlings are greatly improved.

(7) After the transplanting in the greenhouse is completed for one week, the kraft paper bags are rotten and fall into the soil, and the environmental pollution is reduced.

(8) The method has the characteristics of repeated implementation and strong reproducibility; the direct transplanting technology of the arched plastic greenhouse, the special nutrient soil and nutrient solution and the kraft paper bag is adopted, the outdoor seedling hardening and rejuvenating step in the conventional transplanting is omitted, the mechanical damage in the seedling transplanting is reduced, the cost can be saved by 10-20%, the greenhouse has the advantages of sunlight irradiation, constant temperature, proper humidity, good ventilation, insect damage prevention and the like, the haploid seedling growth is facilitated, the survival rate of the young buds can reach about 80-90%, and the corn haploid multiplying power can be improved by about 25-34%.

Drawings

In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings used in the description of the embodiments or the prior art will be briefly described below, it is obvious that the drawings in the following description are only embodiments of the present invention, and for those skilled in the art, other drawings can be obtained according to the provided drawings without creative efforts.

FIG. 1 is a drawing showing the construction of an arched vinyl house according to the invention;

FIG. 2 is the attached drawing of the preparation and filling of the special nutrient soil of the invention;

FIG. 3 is a drawing showing the soil content of the kraft paper bag of the invention;

FIG. 4 is a drawing showing the nondestructive seedling transplanting of kraft paper bags in a shed according to the present invention;

FIG. 5 is a drawing showing the effect of the kraft paper bag of the present invention in direct transplantation, hardening and rejuvenating seedlings;

FIG. 6 is a view showing a plastic greenhouse for heat preservation, moisture preservation, ventilation, sufficient illumination and insect pest prevention according to the present invention;

FIG. 7 is a view showing the micro-spraying of the soil in the greenhouse of the present invention;

FIG. 8 is a schematic view of a conventional transplanting method according to the present invention for constructing a shading net;

FIG. 9 is a drawing showing seedling transplanting in seedling pots in a conventional transplanting method;

FIG. 10 is a schematic view of an outdoor hardening-seedling and rejuvenating seedling of a conventional transplanting method;

FIG. 11 is a drawing showing the seedling hardening and reviving effects of the conventional transplanting method;

FIG. 12 is a drawing showing a seedling transplantation in a conventional transplantation method;

FIG. 13 is a graph showing a comparison of growth vigor of haploid seedlings in direct paper bag transplanting in a greenhouse according to example 1 and in a conventional seedling pot transplanting method according to a comparative example;

wherein, 1: the kraft paper bag of the invention can be directly transplanted to the haploid seedling growth; 2: haploid seedling growth vigor is achieved by a conventional cultivation method of a seedling pot.

Detailed Description

The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.

The special vegetable greenhouse film is purchased from Qingzhou Yuxin Source greenhouse engineering Co., Ltd; the special vegetable greenhouse insect prevention net is purchased from Taizhou city agriculture net company Limited of Zhejiang province; "Huawu" brand nutrient fertilizer is purchased from Shanghai Yongtong ecological engineering, Inc.; kraft paper bags were purchased from new bridge plus large screen mills in the road and bridge area of Taizhou city.

Example 1

A cultivation method for improving doubling efficiency of corn haploid comprises the following specific steps:

(1) building a plastic greenhouse: the appearance of the greenhouse is built in an arched manner, the height in the greenhouse is 1.8-2.0m, the arched part of the greenhouse is covered by a special vegetable greenhouse film which has high light transmittance, heat preservation and wind prevention, the periphery of the greenhouse is built in an enclosing manner by a special vegetable greenhouse insect-proof net with high density, the ventilation and insect prevention in the greenhouse are kept, and the temperature is kept at 28-32 ℃; the construction of the arched plastic greenhouse is shown in figure 1;

(2) preparing special nutrient soil: mixing danish peaty peat soil, perlite and turfy soil according to the proportion of 5: 1: 4, obtaining special nutrient soil for later use; the preparation and filling of the special nutrient soil are shown in figure 2;

(3) preparing nutrient solution and disinfectant: the nutrient fertilizer for the flower-scarless brand is prepared by mixing the following components in parts by weight of 1: 500, and mixing the rooting powder according to the proportion of 1: 200, diluting, and mixing the diluted nutrient fertilizer and the diluted rooting powder in equal volume to prepare a special mixed nutrient solution for later use; mixing the sterilizing agent according to the proportion of 1: 1000, diluting and uniformly stirring to prepare a disinfectant for later use;

(4) seedling preparation: spraying the special nutrient soil prepared in the step (2) by adopting the nutrient solution prepared in the step (3), wherein the special nutrient soil keeps a semi-dry and semi-wet state, and then filling the semi-dry and semi-wet special nutrient soil into a kraft paper bag, wherein the soil filling amount is two thirds of the capacity of the paper bag (figure 3);

(5) transplanting seedlings: taking out the corn haploid seedlings in the laboratory culture bottles, and carefully transferring the corn haploid seedlings into the kraft paper bags in the step (4) according to different numbers;

(6) transplanting in a greenhouse: manually digging a ditch with the depth of 15-20cm, placing kraft paper bags containing corn haploid seedlings into the ditch according to the number (figure 4), manually filling and slightly pressing a small amount of fine soil around the paper bags (figure 5); timely sprinkling irrigation and recording after transplanting is completed;

(7) field management: keeping the temperature in the greenhouse at 28-32 ℃ and the humidity at 60-75%, hardening seedlings for 10-15 days, spraying special nutrient solution and disinfectant once every 2-3 days on the special nutrient soil in the kraft bags, spraying 40-50ml of special nutrient solution and 10-15ml of disinfectant once on each kraft bag, keeping the soil in the kraft bags moist, and fertilizing when the seedlings grow to 6-7 leaves; the field management work is carried out by a specially-assigned person, so that the greenhouse is ensured to have no weeds (figure 6) and sufficient water (figure 7).

And when the seedlings grow to 6-7 leaves, counting the survival rate of the seedlings, counting the number of fructification plants after harvesting, and calculating the multiplying power of the haploids, wherein the results are shown in table 1.

TABLE 1 haploid seedling survival (%) and doubling (%) of each variety using the cultivation method of the present invention

Note: the survival rate (%) of the seedlings is the number of surviving seedlings/total number of transplanted seedlings is 100; haploid multiplying power (%) (number of successfully doubled strains/number of survival seedlings) × 100; doubling the number of successful plants means the number of plants that are fructified at harvest.

Comparative example

The conventional cultivation method comprises the following operation steps: building a shading net, preparing special nutrient soil, preparing nutrient solution and disinfectant, preparing seedling, transplanting seedlings, hardening and reviving seedlings outdoors, transplanting in the field and managing in the field.

The method comprises the following specific steps:

(1) building a shading net: the black shading net with holes is adopted for surrounding construction (figure 8), and the ventilation in the shed is kept.

(2) Preparing nutrient soil: mixing danish peaty peat soil and perlite according to a preparation ratio of 5: 2.

(3) preparing a nutrient solution: the nutrient fertilizer for flowers without flowers comprises the following components in percentage by weight: 500 for dilution.

(4) Seedling preparation: spraying a nutrient solution on nutrient soil prepared in advance, keeping the nutrient soil in a semi-dry and semi-wet state, and then filling the nutrient soil into a seedling pot, wherein the soil filling amount is two thirds of the capacity of the seedling pot.

(5) Transplanting seedlings: the seedlings in the laboratory flasks were removed and carefully transferred to pots with different numbers (FIG. 9).

(6) Outdoor hardening and seedling reviving: the seedling pots are placed on the seedling frame according to the numbers, placed in seed boxes, moved out of the room and placed in a shading net (figure 10). Keeping the temperature in the shade net at 28-32 deg.C and humidity at 60-75%, and hardening seedling for 10-15 days (FIG. 11).

(7) Transplanting in the field: manually digging a ditch with the depth of 15-20cm, putting a seedling pot containing haploid seedlings into the ditch according to the number (figure 12), carefully digging out or pouring the seedlings by hands, transplanting the seedlings into the ditch, manually filling and lightly pressing a small amount of fine soil. And (5) timely sprinkling irrigation and recording after transplanting is finished.

(8) Field management: keeping soil moist, and fertilizing when the seedlings grow to 6-7 leaves. The field management work is carried out by a specially-assigned person, so that the soil is ensured to have sufficient moisture and no weed.

And when the seedlings grow to 6-7 leaves, counting the survival rate of the haploid seedlings of each variety by the conventional cultivation method, counting the number of fruiting plants after harvesting is finished, and calculating the doubling rate of the haploid, wherein the results are shown in table 2.

TABLE 2 haploid seedling survival (%) and doubling (%) of each variety using conventional cultivation method

Note: the survival rate (%) of the seedlings is the number of surviving seedlings/total number of transplanted seedlings is 100; haploid multiplying power (%) (number of successfully doubled strains/number of survival seedlings) × 100; doubling the number of successful plants means the number of plants that are fructified at harvest.

The improved survival rate and doubling rate of seedlings for each variety after comparison of the cultivation method of the present invention (example 1) and the conventional cultivation method (comparative example) are shown in table 3.

TABLE 3 survival (%) and doubling (%) of seedlings of each variety after comparison between the cultivation method of the present invention and the conventional cultivation method

The method of the invention omits the outdoor seedling exercising and reviving process in the conventional cultivation method, and reduces the labor cost. The direct transplanting of the kraft paper bags in the greenhouse is beneficial to rejuvenation, the survival rate and the doubling rate of the haploid seedlings are greatly improved (figure 13), the mechanical damage and the environmental pollution of the seedlings are reduced, the cost of breeding materials is reduced, the operation is simple and convenient, and the large-scale popularization and application can be realized.

The previous description of the disclosed embodiments is provided to enable any person skilled in the art to make or use the present invention. Various modifications to these embodiments will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other embodiments without departing from the spirit or scope of the invention. Thus, the present invention is not intended to be limited to the embodiments shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.