CN108782226A - A kind of method for creating of male sterile wheat line and application - Google Patents

Abstract

The invention belongs to the technical field of wheat breeding, and in particular relates to a creation method and application of a wheat sterile line. In this method, the first step is to plant the wheat seeds normally in the experimental field; the second step is to wait until the internal morphology of the wheat grows to the stage of pistil primordium differentiation, and the external morphology shows that the flag leaf grows to half the length of the second leaf At this time, the high temperature treatment is carried out for 10-12 days, and the temperature of the wheat growth environment is 10-15°C higher than the normal environment temperature, so as to obtain the wheat sterile line. The sterile line created in this way has no substantial damage to the ovary, and experiments have proved that the seed setting rate of artificial pollination is above 96%. By creating a sterile line in this way, to a certain extent, it is possible to freely prepare hybrid combinations among different wheat varieties, thereby further breeding dominant hybrid combinations, and finally breeding hybrid wheat varieties.

Description

A kind of creation method and application of wheat sterile line

technical field

The invention belongs to the technical field of wheat breeding, and in particular relates to a creation method and application of a wheat sterile line.

Background technique

Wheat is a gramineous crop widely planted in the world, and it has always been one of the main varieties of traditional crops in our country. According to statistics, wheat production must increase by 2% per year to meet the needs of people all over the world. In the short term, one of the ways to increase wheat yield is still to breed high-yielding varieties, and the breeding of hybrid varieties is an important way of expressing high-yielding varieties. The routine breeding of wheat sterile lines takes a long time, while the creation of some other sterile lines may depend on weather conditions or be unfriendly to the environment, and the creation cost is relatively high. Therefore, it is an urgent task to choose a more favorable and low-cost way to create sterile lines of wheat. The present invention aims at creating a low-cost convenient wheat sterile line shortcut.

The conventional wheat breeding process is to select excellent lines after natural hybridization or self-crossing until a new variety is bred. However, the conventional breeding method takes a long time and has low efficiency, which limits the development of the wheat breeding process. Therefore, in order to reduce the breeding work, select suitable It is particularly important to have a good hybrid male parent, female parent and suitable hybridization methods. Therefore, the creation of sterile lines becomes the basis for the selection and breeding of hybrid varieties.

With the development of molecular biology technology, transgenic technology is gradually applied in wheat breeding, wheat sterility genes have also been found and cloned, such as MS1 gene, but there is still a long way to go to apply these sterility genes to wheat hybrid breeding, so through A low-cost and non-polluting way to create wheat sterile lines is still the basis of hybrid wheat breeding.

Contents of the invention

The invention provides a method and application for creating a wheat sterile line, which creates a wheat sterile line in a low-cost and pollution-free manner, laying a foundation for hybrid wheat breeding.

The object of the present invention is to provide a kind of creation method of wheat sterile line, comprises the following steps:

In the first step, the wheat seeds are normally planted in the experimental field;

In the second step, when the wheat grows to the internal form of pistil primordia, the external form shows that the flag leaf grows to half the length of the second leaf, at this time, high temperature treatment is carried out for 10-12 days, and the wheat grow The ambient temperature is 10-15°C higher than the normal ambient temperature, and the wheat sterile line is obtained.

Preferably, the high temperature treatment method of the above-mentioned wheat sterile line creation method is as follows: build a shed above the wheat area to be treated at high temperature, then cover the shed roof with a light-transmitting plastic film, and dismantle the shed after 10-12 days.

Preferably, in the above-mentioned method for creating a sterile line of wheat, the plastic film material is polyvinyl chloride resin, and the thickness is 0.12mm.

Preferably, in the creation method of the above-mentioned wheat sterile line, the shed is a square shed with a height of 1.5 meters set up with a steel pipe support.

Preferably, in the above-mentioned method for creating a sterile wheat line, the wheat seeds are planted from October 6th to October 17th.

Preferably, in the above-mentioned method for creating a sterile wheat line, the wheat seeds used in the first step are Xinong 1376 and Zhoumai 28.

The present invention also provides an application of the method for creating a sterile line of wheat in the combination of strong heterosis in wheat.

Compared with the prior art, the creation method and application of the wheat sterile line provided by the present invention have the following beneficial effects:

(1) The present invention creates the mode of high-temperature environment by film covering, develops the new method of establishing wheat sterile line, the wheat of normal growth is processed by the method for high-temperature stress, finds through investigation temperature, the temperature of film covering is generally higher than The normal ambient temperature is 10°C, and the temperature difference between morning and night fluctuates about 10-15°C. The sterile lines created in this way are characterized by smaller anthers and shorter lengths, and finally show no accumulation of starch grains, and the anthers do not dehisce. The tapetum of the male sterile line was degraded ahead of time, and the apoptosis was significantly higher than that of the normal wheat anthers.

(2) The wheat sterile line created by us, the final investigation results show that the self-seed setting rate of wheat variety Xinong 1376 in normal growth is 100%, the self-seed-setting rate of high-temperature treatment is close to 0, and the self-seed-setting rate of pollination hybridization after high-temperature treatment is The seed setting rate of hybridization was 96.3%; Zhoumai 28, the selfing seed setting rate of normal growth was 100%, the selfing seed setting rate of high temperature treatment was close to 0, and the hybrid seed setting rate of pollination hybrid after high temperature treatment was 98.2%. Therefore, we believe that the wheat sterile system can be successfully constructed by high temperature treatment. The method is low-cost and pollution-free, and the induction of sterile wheat through high-temperature treatment provides a convenient and environment-friendly shortcut for creating different hybrid vigor combinations.

Description of drawings

Fig. 1 is a flowchart of the method for creating a sterile line of wheat of the present invention and its effect verification.

Detailed ways

The present invention will be described in detail below in conjunction with specific examples, but should not be construed as a limitation of the present invention. The test methods for which specific conditions are not indicated in the following examples are usually operated under conventional conditions, and the steps are not described in detail because they do not involve the invention point.

A method for creating a wheat sterile line provided by the invention comprises the following steps:

In the first step, the wheat seeds are normally planted in the experimental field;

In the second step, when the wheat grows to the internal form of pistil primordia, the external form shows that the flag leaf grows to half the length of the second leaf, at this time, high temperature treatment is carried out for 10-12 days, and the wheat grow The ambient temperature is 10-15°C higher than the normal ambient temperature, and the wheat sterile line is obtained.

Zhoumai 28 is a semi-winter late-maturing variety with semi-creeping seedlings, strong seedlings, narrow and long leaves, and good cold resistance in winter. Strong tillering ability, medium tillering rate, fast jointing in early spring, fast polarization, late heading, moderate spring cold resistance, late high temperature resistance, and medium maturity. The plant height is 76 cm, the plant type is moderately elastic, and the lodging resistance is good. The ear layer is relatively neat, the internodes under the ear are long, the leaves are flushed, and the stems and leaves are heavy in wax. The ear is nearly oblong, the ear length is sparse, long awn, white shell, white grain, horny kernel, good plumpness, and medium black embryo rate.

Xinong 1376 is an early-maturing variety with semi-spring nature, moderate tillering, high ear formation rate, plant height of 80cm, good yellow shedding, strong comprehensive disease resistance in the field, high grain weight per ear, and high yield potential. White grains, 1000-grain weight about 40g. Xinong 1376 is an ideal supporting variety for intercropping.

Therefore, in the following examples and comparative examples, we used Zhoumai 28 and Xinong 1376 as research objects.

Preferably, a method for creating a wheat sterile line provided by the present invention includes the following examples.

Example 1

A method for creating a wheat sterile line, the method comprises the following steps:

The first step, on October 6, the wheat seeds of Zhoumai 28 and Xinong 1376 were planted in the experimental field normally, and the routine management of watering, fertilizing and weeding was carried out according to the normal wheat cultivation procedures;

In the second step, when the Zhoumai 28 and Xinong 1376 wheat varieties grow to the stage of differentiation of pistil primordium in the internal form, and the external form shows that the flag leaf grows to half the length of the second leaf, high temperature treatment is carried out at this time for 10 days, and the temperature of the wheat growth environment is 10-15°C higher than the normal environment temperature to obtain a wheat sterile line.

The method of high temperature treatment is as follows: build a shed above the area of wheat to be treated with high temperature, and then cover the roof of the shed with a light-transmitting plastic film. In this embodiment 1, a 1.5-meter-high square shed is built with a steel pipe support above the high-temperature-treated wheat area, and the shed is covered with a light-transmitting plastic film (the material is polyvinyl chloride (PVC) resin, with a thickness of 0.12 mm) Top, the film thickness is 0.12mm, that is, covered with a layer of plastic film;

Example 2

A method for creating a wheat sterile line, the method comprises the following steps:

The first step, on October 17, the wheat seeds of Zhoumai 28 and Xinong 1376 were normally planted in the experimental field, and the routine management of watering, fertilizing and weeding was carried out according to the normal wheat cultivation procedures;

In the second step, when the Zhoumai 28 and Xinong 1376 wheat varieties grow to the stage of differentiation of pistil primordium in the internal form, and the external form shows that the flag leaf grows to half the length of the second leaf, high temperature treatment is carried out at this time for 12 days, and the temperature of the wheat growth environment is 10-15°C higher than the normal environment temperature to obtain a wheat sterile line.

The method of high temperature treatment is as follows: build a shed above the area of wheat to be treated with high temperature, and then cover the roof of the shed with a light-transmitting plastic film. In this embodiment 2, a 1.5-meter-high square shed is built with a steel pipe support above the high-temperature-treated wheat area, and then the shed is covered with a light-transmitting plastic film (the material is polyvinyl chloride (PVC) resin, and the thickness is 0.12mm). Top, the film thickness is 0.12mm, that is, covered with a layer of plastic film;

Xinong 1376 is later than Zhoumai 28,

Comparative example 1

The first step, on October 6, the wheat seeds of Zhoumai 28 and Xinong 1376 were normally planted in the experimental field, and the routine management of watering, fertilizing and weeding was carried out according to the normal wheat cultivation procedures;

In the second step, when the Zhoumai 28 and Xinong 1376 wheat varieties grow to the stage of differentiation of pistil primordium in the internal form, and the external form shows that the flag leaf grows to half the length of the second leaf, at this time, the wheat is not treated. Do any treatment, still adopt conventional planting method, get normal wheat plant.

In order to illustrate the beneficial effects of the method of the present invention, we have carried out the following experiments.

Fig. 1 is the flowchart of the creation method and effect verification of the wheat sterile line of the present invention, we first carry out the normal planting of the wheat variety, wait until the internal form is the differentiation stage of the pistil primordium, and the external form shows that the flag leaf grows to the inverted 1/2 length of two leaves, then carry out high temperature treatment and normal growth treatment according to the method of embodiment 1 and comparative example 1 respectively. Then the wheat growth temperature, anther quality, self-pollination seed-setting rate and artificial pollination seed-setting rate were tested respectively.

1. Wheat growth temperature:

The normal growing wheat was treated by the high temperature treatment method in Example 1. After 10 days of continuous temperature investigation, it was found that the temperature covered by the film was generally 10° C. higher than the normal ambient temperature in Comparative Example 1.

2. Anther quality

The anther quality test steps are as follows: (1) Take Xinong 1376 and Zhoumai 28 wheat anthers in different stages of normal growth and high temperature treatment, measure the length of wheat anthers with a vernier caliper, and record them respectively. Among them, the different stages refer to the differentiation stage of pistil primordium, tetrad stage, early mononucleate stage, late stage mononucleate stage, dinucleate stage, and trinucleate stage. Take samples for normal growth and high temperature treatment respectively.

(2) After measuring the length of wheat anthers, store them in FAA fixative solution (the formula of FAA fixative solution is as follows: 5 mL of formaldehyde solution with a volume fraction of 38, 5 mL of glacial acetic acid, 90 mL of ethanol solution with a volume fraction of 70%, and 5 mL of glycerin), and then follow the routine sectioning procedures The detection of paraffin section and apoptosis were recorded and observed respectively. The results showed that the sterile line created by the method in Example 1 specifically showed that the anthers became smaller and shorter, and finally showed no accumulation of starch grains, and the anthers did not dehisce. The tapetum of the sterile line was degraded ahead of time, and the apoptosis was significantly higher than that of the normal wheat anthers in Comparative Example 1.

3. Self-fertilization rate:

Bagging treatment comparative example 1: 30 pieces of Xinong 1376 wheat with normal growth, check the self-seed setting rate of normal wheat when harvesting; Bagging treatment comparison example 1: 30 wheat with normal growth Zhoumai 28, check the normal wheat when it is to be harvested self-fertilization rate. 30 Xinong 1376 wheat grown in bagging treatment example 1 at high temperature, check the self-seed setting rate of wheat induced by high temperature when waiting to be harvested; 30 Zhoumai 28 wheat grown in bagging treatment example 1 at high temperature, check when waiting to harvest Wheat self-seed setting rate induced by high temperature.

The final investigation results show that the selfing seed setting rate of wheat variety Xinong 1376 under normal growth is 100%, and that under high temperature treatment is close to 0; Zhoumai 28, the selfing seed setting rate under normal growth is 100%, and the selfing seed setting rate under high temperature treatment is 100%. The seed setting rate is close to 0.

4. Artificial pollination seed setting rate

The ears are trimmed according to the conventional hybridization treatment method, that is, half of the awns and glumes of the wheat ears after the high temperature treatment are cut off with scissors, and then pollinated with normal pollen and bagged. Co-process 30 ears of Xinong 1376 wheat grown at high temperature in Example 1, check the artificial pollination hybridization seed setting rate of high-temperature treatment when harvesting; co-process 30 ears of Zhoumai 28 wheat grown at high temperature, check high-temperature treatment when harvesting seed setting rate of artificial pollination hybrids. The results showed that the hybrid seed setting rate of the pollination cross of wheat variety Xinong 1376 after high temperature treatment was 96.3%, and the hybrid seed setting rate of Zhoumai 28 pollination cross after high temperature treatment was 98.2%.

In summary, we confirmed that the wheat anthers treated with high temperature were completely aborted, and the wheat sterile system could be successfully constructed, while the wheat ovary treated with high temperature was not affected in any way. The induction of sterile wheat by high temperature treatment provides a convenient and environment-friendly shortcut to create different combinations of hybrid vigor.

It should be noted that when a numerical range is involved in the present invention, unless otherwise specified in the present invention, the two endpoints of each numerical range and any value between the two endpoints can be selected. In order to avoid redundant description, the present invention describes preferred embodiments and effects. Although the preferred embodiments of the present invention have been described, those skilled in the art can make these embodiments once they know the basic inventive concepts. Additional changes and modifications. Therefore, it is intended that the appended claims be construed to cover the preferred embodiment as well as all changes and modifications which fall within the scope of the invention.

Obviously, those skilled in the art can make various changes and modifications to the present invention without departing from the spirit and scope of the present invention. Thus, if these modifications and variations of the present invention fall within the scope of the claims of the present invention and equivalent technologies thereof, the present invention also intends to include these modifications and variations.

Claims (7)

1. a kind of method for creating of male sterile wheat line, which is characterized in that include the following steps：

The first step normally plants wheat seed in experimental plot；

Second step waits for that wheat growth to external morphology is the Pistil And Stamen former base idiophase, and formalness shows as boot leaf and grows to down The half length of two leaves, carries out high-temperature process 10~12 days at this time, and the temperature of wheat growth environment is made to compare normal environment Temperature is higher by 10~15 DEG C, obtains male sterile wheat line.

2. the method for creating of male sterile wheat line according to claim 1, which is characterized in that the method for high-temperature process is as follows： In the wheat overlying regions scaffolding for waiting for high-temperature process, the covered rearing with plastic film ceiling of light transmission is then used, canopy is removed after 10~12 days.

3. the method for creating of male sterile wheat line according to claim 2, which is characterized in that the plastic film material is poly- Vinyl chloride resin, thickness 0.12mm.

4. the method for creating of male sterile wheat line according to claim 2, which is characterized in that canopy is set up with steel pipe support 1.5 meters high of rectangular canopy.

5. the method for creating of male sterile wheat line according to claim 1, which is characterized in that the implantation time of wheat seed is 6~October 17 October.

6. the method for creating of male sterile wheat line according to claim 1, which is characterized in that used wheat in the first step Seed is western agriculture 1376 and all wheats 28.

7. application of the method for creating of male sterile wheat line according to claim 1 in wheat hybridizing influential point set.