CN114128706B

CN114128706B - Pineapple pollination liquid and hybrid pollination method

Info

Publication number: CN114128706B
Application number: CN202111423935.7A
Authority: CN
Inventors: 杨祥燕; 蔡元保
Original assignee: Guangxi Subtropical Crops Research Institute
Current assignee: Guangxi Subtropical Crops Research Institute
Priority date: 2021-11-26
Filing date: 2021-11-26
Publication date: 2022-06-10
Anticipated expiration: 2041-11-26
Also published as: CN114128706A

Abstract

The invention relates to the technical field of pineapple breeding, and particularly discloses a pineapple pollination liquid which comprises the following components in parts by mass: 0.012-0.03% propyl gallate, 0.02-0.035% betaine, 8-12% chitosan oligosaccharide, 3-6% mannitol, 0.05-0.08% xanthan gum, 0.04-0.07% borax and the balance water. The pineapple pollination liquid has the function of permeation regulation, reduces the absorption speed of pollen to water, further regulates the water potential in the pollen and prevents the pollen from swelling and dying in the water solution; the pollen has good suspension dispersibility in the pollination liquid; can keep the activity of pollen, improve the activity and the resistance of stigma, is favorable for the stable attachment of the pollen to the stigma, promotes the germination of the pollen, thereby improving the pollination effect and the pollination success rate.

Description

Pineapple pollination liquid and hybrid pollination method

Technical Field

The invention belongs to the technical field of pineapple breeding, and particularly relates to pineapple pollination liquid and a hybrid pollination method.

Background

Pineapple is also called pineapple, is one of four tropical fruits in the world, has the characteristics of sour, sweet, delicious taste, rich nutrition and the like, can be directly eaten, can be processed for the second time to prepare various products, has strong adaptability to pineapple plants, is barren-resistant, drought-resistant, less in plant diseases and insect pests, easy to cultivate, high in yield and the like, and has high economic benefit. In recent years, the pineapple planting area and yield in China have a steady rising trend, and the pineapple industry has become one of the important economic prop industries of the hotspot agriculture in China. The Chinese pineapple has limited resource and single structure, the main cultivated variety is Bali at present, and the main cultivated variety is acaine without thorns, the two varieties account for about 90 percent of the Chinese pineapple, and the serious seed degeneration, the yield and the quality are reduced. The variety is single, the main cultivated variety is degenerated, and the main factor seriously restricts the healthy development of the pineapple industry in China. There is an urgent need to develop new varieties to overcome the above problems. At present, the pineapple selective breeding work in China is in the stage of introduction and selective breeding, and has a larger gap compared with the directional breeding research of the main country of pineapples abroad.

The pineapple has the characteristic of self-blooming, and the pollination among the same variety or the same type of varieties is not full or the seed setting rate is low, so that the excellent parents of different lines are selected for hybridization or distant hybridization, and a large amount of hybrid seeds and a large range of variation can be obtained. The artificial supplementary pollination is an important step for obtaining hybrid seeds of pineapples, and the current artificial supplementary pollination mode mainly comprises the following steps: firstly, manually ordering, namely collecting pollen by using tweezers, then dipping the pollen by using a coarse tweezers or a brush pen, coating the pollen on a stigma to pollinate, wherein the mode has the defects of easy damage to the stigma and the pollen, easy loss of the pollen activity germination rate, time and labor consumption in the operation process, low efficiency, high labor cost and the like; secondly, the manual auxiliary solid powder spraying mode has the problems of large pollen dosage, high cost, small pollen adhesion on the flower column and the like; and thirdly, artificial supplementary liquid pollination, which is a novel pollination technology, has the advantages of high pollination speed, labor saving and the like, can increase the water and nutrient of the stigma and prolong the stigma pollination fertilization time. However, the liquid pollination technology is still immature, and the problems of poor suspension capability of the nutrient solution, uneven pollen dispersion, pollen swelling death in the nutrient solution, low pollen activity and the like still exist. The blooming time of the pineapples of different varieties and different seedling stages is different, the problem of dissatisfaction of the florescence of parents exists, or pollination cannot be carried out in time due to climate conditions or regional isolation, so that the pollen is preserved by adopting reasonable measures, so that the stability and the activity of the pollen are beneficial to improving the success rate of hybrid pollination. The pollination liquid and the hybrid pollination method for pineapple hybrid pollination are developed, the success rate of pineapple pollination is improved, and the method has important significance for pineapple breeding.

Disclosure of Invention

The invention aims to provide a pineapple pollination liquid and a hybrid pollination method, which can maintain pollen activity, promote pollen germination and improve pollination efficiency and success rate.

In order to achieve the purpose, the invention provides a pineapple pollination liquid which comprises the following components in parts by mass: 0.012-0.03% propyl gallate, 0.02-0.035% betaine, 8-12% chitosan oligosaccharide, 3-6% mannitol, 0.05-0.08% xanthan gum, 0.04-0.07% borax and the balance water.

Preferably, in the above pineapple pollination liquid, the pineapple pollination liquid consists of the following components in parts by mass: propyl gallate 0.02%, betaine 0.03%, chitosan oligosaccharide 10%, mannitol 4%, xanthan gum 0.06%, borax 0.05%, and water in balance.

In the pineapple pollination liquid, propyl gallate and mannitol can eliminate free radicals generated in pollen and keep the activity of the pollen; mannitol and betaine have the function of regulating cell penetration, can reduce the absorption speed of pollen to water, further regulate and control the water potential in the pollen to prevent the pollen from swelling and dying, and betaine also has the functions of improving plant resistance, stimulating female parent flower columns to improve the activity and resistance of the flower columns and increasing the adsorption of the flower columns to the pollen; the chitosan oligosaccharide has positive charge, enhances the adhesion of pollen on the flower column through electrostatic action, also has antibacterial effect and enhances the cell activity, and promotes the germination of pollen tube; the components of the pineapple pollination liquid act synergistically to finally improve the success rate of pineapple pollination.

A hybrid pollination method for pineapples comprises the following steps:

(1) collecting pollen at the flowering phase of the pineapple male parent, and placing the pollen into a pollen storage box for low-temperature storage;

(2) adding the pollen obtained in the step (1) into the pineapple pollination solution of

claim

1 or 2, and uniformly mixing to prepare a pollen suspension with the pollen concentration of 1.2-1.8 g/L;

(3) and (3) spraying the pollen suspension obtained in the step (2) on the stigma of the female pineapple flower after the female pineapple plant enters the flowering phase, and completing cross pollination.

Preferably, in the above pineapple cross pollination method, the pollen storage box comprises a box body and a sealed box cover, the side wall of the box body comprises an inner side wall, a middle accommodating cavity and an outer side wall, the inner side wall is of a porous net structure, the top of the middle accommodating cavity is open, and one or more layers of absorbent fiber paper are laid at the bottom in the box body.

Preferably, in the above method for cross pollination of pineapple, the upper surface of the uppermost layer of the absorbent fiber paper is treated as follows: spraying 1% matrine alcohol solution on the upper surface of the absorbent fiber paper, and then blowing and drying at 40-50 ℃; the dosage of 1% matrine alcohol solution per square meter of absorbent fiber paper is 0.1-0.2 ml, and the 1% matrine alcohol solution is diluted by 30 times of ethanol for spraying. The matrine is attached to the surface of the water-absorbing fiber paper after surface treatment, and can effectively kill harmful microorganisms and small insects in pollen without influencing the activity of the pollen, so that the pollen is prevented from being damaged by the harmful microorganisms and the small insects and losing the activity.

Preferably, in the above method for cross pollination of pineapple, the specific process of step (1) is: collecting mature pollen when flowers on the pineapple male parent inflorescence bloom to the middle, sieving the pollen by a 40-50 mesh sieve, placing the sieved pollen on absorbent fiber paper in a pollen storage box body, and placing the storage box body in a 25-28 ℃ forced air drying box for drying for 1-2 hours; then, filling an oxygen scavenger in the middle accommodating cavity of the side wall of the pollen preservation box, covering the sealing box cover, placing the pollen preservation box in an environment at 10 ℃ for 1-2 hours, and then placing the pollen preservation box for preservation at 4 ℃. The pollen is placed in the pollen storage box for low-temperature storage, the water-absorbing fiber paper in the pollen storage box has high water absorption, the water content in the pollen is favorably accelerated to be reduced, the pollen drying time is reduced, the oxygen concentration in the pollen storage box is reduced through an oxygen scavenger, the pollen metabolism speed is reduced, and the pollen activity time is prolonged.

Preferably, in the step (3) of the pineapple cross pollination method, pollination is carried out after 3 to 5 days after the pineapple female parent plant enters the flowering phase, the pollination time is in clear weather and no dew is between eight and ten am, the pollen is sprayed on the small flowers opening in the inflorescence, and the pollen is sprayed on the stigma once every 2 to 3 days and is carried out for 3 to 5 times in total; the use amount of the pollen suspension of each pineapple plant in each pollination is 1.5-2 ml.

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

1. the pineapple pollination liquid has the function of permeation regulation, reduces the absorption speed of pollen to water, further regulates the water potential in the pollen and prevents the pollen from swelling and dying in the water solution; the pollen has good suspension dispersibility in the pollination liquid; can keep the activity of pollen, improve the activity and the resistance of stigma, be favorable to pollen to stably attach to stigma, promote the germination of pollen, improve the pollination effect, thereby enhancing the pollination success rate.

2. In the pineapple hybrid pollination method, pollen is stored at low temperature through the pollen storage box, so that the activity of pollen of the male parent can be guaranteed, and the pollen utilization rate and the hybridization success rate are improved; pollen is added into the pollination liquid to prepare a pollen suspension, and the pollen suspension is sprayed for multiple times of pollination and the spraying amount is controlled to improve the probability of the female parent florets receiving the pollen of the male parent and improve the success rate of hybridization.

3. The method for cross pollination of pineapples has the advantages of simple operation, less labor, low labor cost, high pollination efficiency, high seed yield, little influence of natural environment conditions, and capability of overcoming the problems of poor pollination and the like caused by flowering asynchronism or incapability of timely pollination under climatic conditions.

Drawings

Fig. 1 is a schematic sectional view showing a pollen storage box according to embodiment 1 of the present invention.

Description of the main reference numerals: the sealing structure comprises a cover body 1, a sealing box cover 2, absorbent fiber paper 3, an outer side wall 4, a middle accommodating cavity 5, an inner side wall 6 and a sealing ring 7.

Detailed Description

The following detailed description of specific embodiments of the invention is provided, but it should be understood that the scope of the invention is not limited to the specific embodiments.

Example 1

The pineapple pollination liquid consists of the following components in percentage by mass: propyl gallate 0.02%, betaine 0.03%, chitosan oligosaccharide 10%, mannitol 4%, xanthan gum 0.06%, borax 0.05%, and water in balance.

A hybrid pollination method for pineapples comprises the following steps:

(1) collecting mature pollen when small flowers on the pineapple male parent inflorescence bloom to the middle, sieving the pollen by a 40-50 mesh sieve, and placing the pollen into a pollen storage box for low-temperature storage;

(2) adding the pollen in the step (1) into the pineapple pollination liquid, and uniformly mixing to prepare a pollen suspension with the pollen concentration of 1.5 g/L;

(3) pollinating 3-5 days after the pineapple female parent plant enters the flowering phase, wherein the pollination time is that the weather is fine and no dew exists between eight and ten am, spraying pollen suspension on the florets opened in the inflorescence to spray pollen on the stigma, and spraying the pollen once every 2-3 days for 3-5 times; the amount of pollen suspension in each pineapple plant is 1.8ml per pollination. Pollinating at intervals of 2-3 days, namely, not influencing the florets after the previous pollination to finish fertilization, and simultaneously increasing more florets to receive male parent pollen to improve the maturing rate.

As shown in fig. 1, the pollen storage box comprises a box body 1 and a sealing box cover 2, wherein the side wall of the box body 1 consists of an outer side wall 4, a middle accommodating cavity 5 and an inner side wall 6, the inner side wall 6 is of a porous net structure, the top of the middle accommodating cavity 5 is provided with an opening for filling a deoxidant, and two layers of absorbent fiber paper 3 are laid at the bottom in the box body 1 and can absorb moisture to accelerate the reduction of the moisture content in pollen; the sealing box cover 2 is internally provided with a rubber sealing ring 7 for enhancing the sealing effect of the pollen preservation box. The thickness of the absorbent fiber paper is 1mm, and the water absorption capacity is 400g/m²The upper surface of the uppermost layer of absorbent fiber paper in the box body 1 is treated as follows: spraying 1% matrine alcohol solution on the upper surface of absorbent fiber paper, and then drying by blowing at 50 deg.C; the dosage of 1% matrine alcohol solution per square meter of absorbent fiber paper is 0.15ml, and the 1% matrine alcohol solution is diluted with 30 times of ethanol for spraying.

The specific process of pollen preservation is as follows: placing the sieved pollen on the uppermost layer of absorbent fiber paper 3 in the box body 1 of the pollen preservation box, and placing the box body 1 of the preservation box in a 28 ℃ blast drying oven for drying for 1 h; then filling an oxygen scavenger in the middle accommodating cavity 5 in the side wall of the pollen preservation box, covering the sealing box cover, placing the pollen preservation box in an environment at 10 ℃ for 2h, and then placing the pollen preservation box in a refrigerator at 4 ℃ for preservation. Firstly drying for 1h to reduce the water content of the pollen, then filling an oxygen scavenger in a pollen preservation box, sealing, and placing in an environment at 10 ℃ for 2h to reduce the oxygen solubility of the pollen preservation box, reduce the metabolism speed of the pollen and prolong the pollen activity time.

Example 2

The pineapple pollination liquid consists of the following components in percentage by mass: propyl gallate 0.024%, betaine 0.028%, chitosan oligosaccharide 9%, mannitol 5%, xanthan gum 0.07%, borax 0.06%, and water in balance.

A hybrid pollination method for pineapples comprises the following steps:

(1) collecting mature pollen when the small flowers on the pineapple male parent inflorescence bloom to the middle, sieving the pollen by a 40-50 mesh sieve, and placing the pollen into a pollen storage box for low-temperature storage, wherein the storage method is the same as that in the embodiment 1;

(3) pollinating 3-5 days after the pineapple female parent plant enters the flowering phase, wherein the pollination time is between eight and ten morning hours when the weather is fine and no dew exists, spraying pollen on stigma by aiming at the small flowers opened in the inflorescence, and spraying the pollen once every 2-3 days for 3-5 times; the amount of pollen suspension in each pineapple plant is 1.8ml per pollination.

Example 3

The pineapple pollination liquid consists of the following components in percentage by mass: propyl gallate 0.015%, betaine 0.032%, chitosan oligosaccharide 11%, mannitol 3.5%, xanthan gum 0.055%, borax 0.04%, and water in balance.

A hybrid pollination method for pineapples comprises the following steps:

(2) adding the pollen in the step (1) into the pineapple pollination liquid, and uniformly mixing to prepare a pollen suspension with the pollen concentration of 1.6 g/L;

Comparative example 1

The difference from the embodiment 1 is that: the pineapple pollinator does not contain propyl gallate, and other parameters and steps are the same as those in example 1.

Comparative example 2

The difference from the embodiment 1 is that: pineapple pollination solution does not contain betaine, and other parameters and steps are the same as example 1.

Comparative example 3

The difference from the embodiment 1 is that: pineapple pollination solution does not contain chitosan oligosaccharide, and other parameters and steps are the same as those in example 1.

Comparative example 4

The difference from the embodiment 1 is that: the pineapple pollination liquid consists of the following components in percentage by mass: 14% of sucrose, 0.06% of xanthan gum, 0.05% of borax and the balance of water, and other parameters and steps are the same as those in example 1.

Test example 1 pollen preservation test

The test subjects were: mature pollen of pineapple

The test method comprises the following steps: fresh mature pollen is collected and divided into 4 equal parts, which are set into A, B, C, D groups, and the pollen is respectively subjected to 4 preservation treatments: group A was performed according to the pollen storage box and storage procedure provided in example 1, group B was performed by placing pollen in a petri dish, drying by blowing at 28 deg.C for 4h, placing in a centrifuge tube, and storing at 4 deg.C; the preservation steps of the group C and the group A are the same, except that the upper surface of the uppermost absorbent fiber paper in the pollen preservation box is not treated; group D was the same as group A except that the pollen storage box was not filled with an oxygen scavenger. A few pollen was taken out of 4 treatment groups and stored for 0 day, 2 days, 5 days, 10 days and 15 days, and the germination rate of the pollen was measured by a medium germination method, and the results are shown in Table 1.

TABLE 1 pollen germination Rate for each treatment group

As can be seen from the table 1, the pollen preservation method provided by the invention can keep the pollen vitality, the germination rate of the pineapple pollen is more than 85% after 10 days of preservation, and the method can be used as a short-term preservation method of the pineapple pollen and is beneficial to hybridization breeding of pineapples.

Experimental example 2 hybrid pollination of pineapple

Performing a pineapple cross pollination test in a pineapple planting base of Guangxi subtropical crop research institute, adopting Bali and caine as experimental objects, and planting the caine one month in advance to enable the flowering phases of the Bali and the caine to be similar; the method comprises the steps of taking Bali as a female parent and cocaine as a male parent, collecting mature pollen when flowers bloom to the middle on a pineapple male parent inflorescence, preserving at low temperature for later use, dividing female parent pineapple plants into 7 groups in total, namely, groups of examples 1-3 and comparative examples 1-4 respectively, beginning to pollinate 10 female parent pineapples in each group when the female parent pineapple plants enter a flowering period of 3 days, weighing the pollen and corresponding pineapple pollination liquid respectively, mixing uniformly to obtain pollen suspension, carrying out spray pollination, pollinating 1 time every 2 days, and pollinating 4 times in total. Then, the number of flowers successfully pollinated is investigated, the number of seeds is investigated after harvesting, and the seed rate is calculated.

TABLE 2 pollination Effect of treatment groups for hybrid pollination

Table 2 shows the pollination effect of the cross pollination of each treatment group, and the cross pollination performed by adopting the cross pollination method provided by the invention has the advantages of good pollination effect, high pollination success rate and seed obtaining rate, and capability of improving the cross breeding efficiency.

The foregoing descriptions of specific exemplary embodiments of the present invention have been presented for purposes of illustration and description. It is not intended to limit the invention to the precise form disclosed, and obviously many modifications and variations are possible in light of the above teaching. The exemplary embodiments were chosen and described in order to explain certain principles of the invention and its practical application to enable one skilled in the art to make and use various exemplary embodiments of the invention and various alternatives and modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the claims and their equivalents.

Claims

1. The pineapple pollination liquid is characterized by comprising the following components in percentage by mass: 0.012-0.03% propyl gallate, 0.02-0.035% betaine, 8-12% chitosan oligosaccharide, 3-6% mannitol, 0.05-0.08% xanthan gum, 0.04-0.07% borax and the balance water.

2. The pineapple pollination liquid as claimed in claim 1, which is characterized by comprising the following components in percentage by mass: propyl gallate 0.02%, betaine 0.03%, chitosan oligosaccharide 10%, mannitol 4%, xanthan gum 0.06%, borax 0.05%, and water in balance.

3. A hybrid pollination method of pineapples is characterized by comprising the following steps:

(2) adding the pollen obtained in the step (1) into the pineapple pollination solution of claim 1 or 2, and uniformly mixing to prepare a pollen suspension with the pollen concentration of 1.2-1.8 g/L;

4. The hybrid pollination method for pineapples according to claim 3, wherein the pollen storage box comprises a box body and a sealing box cover, the side wall of the box body comprises an inner side wall, a middle containing cavity and an outer side wall, the inner side wall is of a porous net structure, the top of the middle containing cavity is open, and one or more layers of absorbent fiber paper are laid at the bottom in the box body.

5. The method for cross-pollination of pineapple as claimed in claim 4, wherein the upper surface of the uppermost absorbent fiber paper is treated by: spraying a 1% matrine alcohol solution on the upper surface of the absorbent fiber paper, and then drying by blowing at 40-50 ℃; the dosage of 1% matrine alcohol solution per square meter of absorbent fiber paper is 0.1-0.2 ml, and the 1% matrine alcohol solution is diluted by 30 times of ethanol for spraying.

6. The hybrid pollination method for pineapple according to claim 5, wherein the specific process of the step (1) is as follows: collecting mature pollen when flowers on the pineapple male parent inflorescence bloom to the middle, sieving the pollen by a 40-50 mesh sieve, placing the sieved pollen on absorbent fiber paper in a pollen storage box body, and placing the storage box body in a 25-28 ℃ forced air drying box for drying for 1-2 hours; then filling an oxygen scavenger in the middle accommodating cavity of the side wall of the pollen preservation box, covering the sealing box cover, placing the pollen preservation box in an environment at 10 ℃ for 1-2 hours, and then preserving at 4 ℃.

7. The hybrid pollination method for pineapples according to claim 3, wherein in the step (3), pollination is carried out after 3 to 5 days after the pineapple female parent plant enters the flowering phase, the pollination time is in clear weather and between eight and ten am without dew, the pollen is sprayed on the stigma by aiming at the small flowers opening in the inflorescence, and the pollination is carried out once every 2 to 3 days for 3 to 5 times; the use amount of the pollen suspension of each pineapple plant in each pollination is 1.5-2 ml.