CN112493114B - Glue solution hybrid pollination method - Google Patents
Glue solution hybrid pollination method Download PDFInfo
- Publication number
- CN112493114B CN112493114B CN202011188038.8A CN202011188038A CN112493114B CN 112493114 B CN112493114 B CN 112493114B CN 202011188038 A CN202011188038 A CN 202011188038A CN 112493114 B CN112493114 B CN 112493114B
- Authority
- CN
- China
- Prior art keywords
- pollen
- glue solution
- solution
- culture solution
- culture
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
Images
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05D—INORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C; FERTILISERS PRODUCING CARBON DIOXIDE
- C05D9/00—Other inorganic fertilisers
- C05D9/02—Other inorganic fertilisers containing trace elements
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05F—ORGANIC FERTILISERS NOT COVERED BY SUBCLASSES C05B, C05C, e.g. FERTILISERS FROM WASTE OR REFUSE
- C05F11/00—Other organic fertilisers
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
-
- C—CHEMISTRY; METALLURGY
- C05—FERTILISERS; MANUFACTURE THEREOF
- C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
- C05G5/00—Fertilisers characterised by their form
- C05G5/20—Liquid fertilisers
Abstract
The invention discloses a glue solution hybrid pollination method, and belongs to the technical field of hybrid breeding. The method comprises the following steps: (1) preparing male parents and female parents; (2) sleeving a pollen isolation net; (3) preparing a glue solution culture solution; (4) preparing a glue solution pollen culture solution; (5) treating the stamens of the female parent; (6) and (5) cross pollination. The method is simple and convenient to operate, and solves the problems that manual emasculation is slow, small flowers are difficult to emasculate, flowers are easy to hurt in the emasculation process, the normal development of fruits is influenced and the like; the problem that pollen is not easy to attach to the pistil stigma is solved, the pistil stigma can be prevented from being influenced by the external environment, the success rate of hybrid pollination is effectively improved, and the purpose of improving the fruit setting rate is achieved. The method is suitable for the majority of plants to carry out cross breeding, effectively reduces the working intensity of cross breeding, improves the working efficiency of cross breeding, and has great significance for accelerating the process and efficiency of cross breeding of special variety plants.
Description
Technical Field
The invention belongs to the technical field of cross breeding, and particularly relates to a glue solution cross pollination method.
Background
The cross pollination is the most traditional but most direct and effective technical means for breeding new plant varieties, common pollination comprises blast pollination in rice cross breeding, oscillation pollination in corn cross breeding, brush point pollination in common melon and fruit cross breeding and the like, and the conventional pollination methods play an important role in the breeding work of male and female cross-plants or isosexual plants and self-sterile plants. However, when some special plants are self-pollinated by the same flowers and the same flowers, the stamens are short and small, the stamens are tiny and the like, so the operations of emasculation, pollination and the like are difficult to be carried out by using the conventional pollination method. In recent years, people explore a liquid pollination technology, can effectively improve the fruit setting rate of kiwi fruits, pears and the like and improve the fruit quality, but cannot meet the technical requirements in cross breeding. Therefore, the exploration of a practical and efficient hybrid pollination technology becomes a key for overcoming the technical bottleneck of cultivating new varieties of special plants.
The traditional cross breeding and pollination method comprises the following steps: firstly bagging the flowers of the male parent before blooming to protect the purity of pollen, castrating the female parent, removing stamen anther by using scissors and bagging, opening the bag sleeved on the flowers of the male parent when blooming, collecting the pollen, opening the bag sleeved on the flowers of the female parent, uniformly smearing the pollen on the heads of the stamen anther, and then continuously bagging. The manual work workload of castrating is big, takes time and labor, and the shortcoming is many, and some flowers are hardly castrated for the little stamen short flower in addition, often meet unavoidable problem, if: (1) the manual castration is labor-consuming and time-consuming, the workload is huge, and the efficiency is low; (2) flowers are easy to be injured in the castration operation process, normal development of fruits is influenced, self-pollination is easy to be caused if castration is not clean and timely, and hybridization rate is influenced; (3) pollen is not easy to adhere when pistil is fine and smooth. The above problems severely restrict the progress and efficiency of cross breeding of these particular plants.
Disclosure of Invention
The invention aims to provide a glue solution cross pollination method which is used for improving the success rate of cross pollination, reducing the working strength of cross pollination breeding, improving the working efficiency of cross breeding and accelerating the process and efficiency of cross breeding of special variety plants.
The purpose of the invention is realized by the following technical scheme:
a glue solution hybrid pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent varieties which have complementary advantages and similar flowering phases;
(2) sleeving a pollen isolation net: selecting male parent and female parent plants which contain buds to be released or have pistils to be exposed, and respectively covering the male parent and the female parent plants by using a pollen isolation net;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing, standing and refrigerating;
(4) preparing a liquid jelly pollen culture solution: collecting pollen with strong activity on male parent, adding pollen and boric acid into the liquid jelly culture solution of 1/2, and mixing to obtain liquid jelly pollen culture solution;
(5) treatment of maternal stamens: injecting the liquid jelly culture solution into the male core layer of female parent to wrap the anther with the liquid jelly culture solution;
(6) and (3) cross pollination: when the female parent flowers bloom or the pistil is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the pistil stems.
As a further preferred aspect of the present invention, the glue solution culture solution in step (3) comprises 8-10% of sucrose, 1-8% of a gelling agent, 0.5-2% of sodium gluconate, 0.5-2% of a thickening agent, 0.3-0.8% of vitamin C, 0.5-1% of vitamin E, and the balance of water.
As a further preference of the invention, the gelling agent is 2-8% of soluble starch or 1-4% of agar; the viscous agent is sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose.
As a further preferred aspect of the present invention, the preparation of the glue solution culture solution is: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding a thickening agent, uniformly mixing, adding a gelling agent for dissolving, then adding cane sugar for dissolving, finally adding vitamin C and vitamin E, and uniformly mixing to obtain a glue solution culture solution.
In a further preferred embodiment of the present invention, the colloidal solution pollen culture solution of step (4) contains 0.05-0.2 wt% of pollen of male parent and 0.005-0.02 wt% of boric acid.
In a further preferred embodiment of the present invention, the liquid jelly pollen culture medium is used as it is.
In a further preferred embodiment of the present invention, the refrigerating temperature of the liquid culture medium in the step (3) is 3-5 ℃.
In a further preferred embodiment of the present invention, the collecting of pollen in step (4) is performed by dropping pollen on a culture dish with a brush.
As a further optimization of the invention, in the step (2), when the plant height is more than or equal to 150cm and less than or equal to 250cm, a shed type pollen isolation net cover is used; when the plant height is more than 250cm or less than 150cm, the flower-bearing branches or the whole plant are sleeved by the bag-type pollen isolation mesh bag; when flowers are born on the trunk of tall trees, the trunk flowers are wrapped by a sheet-type pollen isolation net.
As a further preferred of the present invention, the pollen isolation net in the step (2) is an ultra-dense nylon net with a pore size of 500-1000 meshes.
The technical principle of the technical scheme of the invention is as follows:
boric acid has an influence on the plant reproduction process, the boron content in plant organs is highest by flowers, stigma and ovary tissues are highest in the flowers, the pollen is low, and boron can stimulate pollen germination, particularly pollen tube elongation; the indirect effect of boron is probably related to the increase of sugar content in nectar and the change of the composition of nectar, so that flowers of insect-vector plants are more attractive to insects; in addition, boric acid disperses pollen in the colloidal pollen culture so that approximately equal amounts of pollen are obtained from each female parent pistil stigma.
The cane sugar not only can provide nutrition for the pollen, but also can adjust osmotic pressure, and effectively prevents the pollen from swelling and cracking.
The vitamin C and the vitamin E can provide nutrition for pollen, and the vitamin C and the vitamin E also have strong antioxidation, can help plants resist the pressure caused by ozone and ultraviolet rays, and have the functions of protecting and delaying the plants; particularly, the vitamin E contains tocopherol, can effectively protect a photosynthetic membrane system rich in unsaturated fatty acid contained in the plant from being damaged by photooxidation, and has good protection effect on the photosynthetic organ of the plant.
The soluble starch is slightly acid or alkali treated starch, and the starch solution has good fluidity when being hot and can form firm and soft gel when being condensed.
Agar has physical and chemical properties such as coagulability and stability, can form complex with some substances, and can be used as thickener, coagulant, suspending agent, emulsifier, antistaling agent and stabilizer.
The sodium carboxymethyl cellulose and the hydroxypropyl methyl cellulose have excellent performances of bonding, thickening, emulsifying, suspending, film forming, colloid protection, moisture retention, enzymolysis resistance, metabolic inertia and the like, and can ensure that a prepared glue solution culture solution is colloidal.
The traditional pollen pollination adopts a pollen suspension, the concentration of sodium carboxymethylcellulose usually adopted by the pollen suspension is 0.01-0.05 percent, the pollen suspension is used for preventing the pollen from rapidly precipitating, but the pollen suspension has low adhesion, is not easy to attach to pistil stigmas and is easy to fall off when shaking occurs. The present invention adopts glue solution culture liquid, and adds sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose with concentration of 0.5-2% as a thickening agent. The concentration adopted by the invention is 50-200 times of the traditional concentration, and the invention aims to ensure that the glue solution culture solution of the invention is colloidal and has strong adhesion, can wrap the anther of the male core layer of the female parent well, and prevents the plants from self-pollination and influencing the hybridization rate. When the protective agent is acted on a colloidal solution pollen culture solution, the pistil stigmas can be well protected, the influence of external environment, such as wind, rain, insect and bird bites and the like, on the success rate of hybrid pollination is avoided, and the pollen can be adhered and directly acted on the pistil stigmas.
The sodium gluconate has good complexing, retarding and water quality stabilizing performances, the prepared glue solution culture solution is colloidal, the situation that the prepared glue solution culture solution is too fast coagulated into a solid substance due to high-content sodium carboxymethylcellulose or hydroxypropyl methylcellulose in the glue solution culture solution is avoided, and the pollen in the glue solution pollen culture solution prepared from the added pollen and boric acid does not lose activity quickly under the condition of long-time use, so that sufficient germination and elongation spaces of the pollen are ensured.
The raw materials and materials used in the invention, such as sucrose, soluble starch, agar, sodium gluconate, sodium carboxymethylcellulose, hydroxypropyl methylcellulose, vitamin C, vitamin E, boric acid, pollen isolation net, injector, dropper, culture dish, brush and the like, can be purchased from the market.
Compared with the prior art, the invention has the advantages and beneficial effects that:
1. the method is simple and convenient to operate, and solves the problems that the artificial emasculation is slow, the small flowers are difficult to emasculate, the flowers are easy to be injured in the emasculation process, the normal development of fruits is influenced and the like by injecting the glue solution culture solution into the stamen layer of the female parent through the injector to wrap the anthers with the glue solution culture solution; the gel solution pollen culture solution is uniformly smeared on the pistil stigma of the female parent, so that the problem that pollen is not easy to attach to the pistil stigma is solved, the pistil stigma can be well protected by the gel solution pollen culture solution, the pistil stigma is prevented from being affected by conditions such as wind, rain, insect and bird bites and the like, the success rate of hybrid pollination is effectively improved, and the purpose of improving the fruit setting rate is achieved. The method is suitable for the hybridization breeding of most plants, improves the success rate of hybridization pollination to a great extent, lightens the working strength of the hybridization breeding, improves the working efficiency of the hybridization breeding and has great significance for accelerating the process and the efficiency of the hybridization breeding of the plants of special varieties.
2. The sucrose in the liquid cement culture solution can provide nutrition for pollen in the pollen germination process, also can play a role in regulating osmotic pressure, maintains normal viability and is beneficial to pollen germination; the pollen and the boric acid are added into the liquid cement culture solution to prepare the liquid cement pollen culture solution, and as boron ions can form a complex with sucrose and sodium gluconate has good complexing performance, sugar is easy to transport in tissues through a plasma membrane, so that the absorption and metabolism of the sugar are promoted, and the boron can participate in the synthesis of pectin substances, thus being beneficial to the construction of pollen tube walls, forming a sugar-boric acid complex by the boron, increasing the absorption of oxygen, combining the boron with the sugar and greatly improving the germination rate of the pollen; the vitamin C and the vitamin E can provide nutrition for pollen, have strong antioxidation, help plants resist the pressure caused by ozone and ultraviolet rays, and have the functions of protecting and delaying the plants; through the action of the nutrient substances, the germination rate of the pollen is greatly improved, and the purpose of improving the fruit setting rate is achieved.
3. The glue solution culture solution and the glue solution pollen culture solution contain high-concentration sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose, and the sodium carboxymethyl cellulose and the hydroxypropyl methyl cellulose have excellent performances of bonding, thickening, emulsification, suspension, film formation, colloid protection, water retention, enzymolysis resistance, metabolic inertia and the like, so that the prepared glue solution culture solution is colloidal, and pollen can be uniformly dispersed in the glue solution pollen culture solution. The glue solution culture solution can well wrap the anther of the male core layer of the female parent, prevent plants from selfing and pollinating, and achieve the purpose of efficient castration; the pollen in the colloidal solution pollen culture solution has good dispersity and stability, and the water-retaining property of the colloidal solution pollen culture solution enables the colloidal solution pollen culture solution to form stable colloid outside the pistil stigma, so that the drying and water loss condition of the colloidal solution pollen culture solution drop on the stigma is avoided, the pollen activity is greatly improved, the pistil stigma can be well protected, and the success rate of hybrid pollination is improved.
Drawings
FIG. 1 is a sectional view of a pistil stigma bud of a yolk fruit.
FIG. 2 is an appearance diagram of a flower bud pollinated by a yolk fruitflower with a liquid culture medium of colloidal pollen.
FIG. 3 is a cut view of a bud of a flower for pollinating an egg yolk fruitflower by the method of the present invention.
FIG. 4 is a schematic view of the bud peeling of Annona squamosa Linn.
FIG. 5 is a diagram of isolated surface peeling of sweetsop flower microcolumn stamen colony culture solution.
FIG. 6 is a cross-sectional view of pollinated gynoecium colloidal pollen culture solution of Annona squamosa Linn.
Note: in order to facilitate the observation of color development, a little ink is added into the glue solution to adjust the color.
Detailed Description
The present invention is further illustrated by the following examples, which are provided only for illustrating the present invention and are not intended to limit the scope of the present invention.
Example 1
A glue solution cross pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent yolk fruit varieties which have complementary advantages and similar flowering phases; the number of the female parent plants and the male parent plants is 8; the height of the egg yolk fruit plant is 260 cm;
(2) covering pollen to isolate the mesh bag: selecting buds (as shown in figure 1) on the lateral branches which are to expose the stigma in afternoon at the speed of 4:30-6:00, removing the buds which are not in the same period, covering the lateral branches with a pollen isolation mesh bag, and binding the lateral branches with a rubber band or a rope, wherein the pollen isolation mesh is an ultra-dense nylon mesh with the aperture of 800 meshes, the diameter opening of the pollen isolation mesh bag covered on the lateral branches of the female parent is 50cm, and the bag depth is 100 cm; the male parent and the female parent are subjected to bagging operation;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing and placing in a refrigerating chamber with the temperature of 4 ℃, standing and refrigerating; the glue solution culture solution comprises 9% of sucrose, 8% of gelling agent, 1% of sodium gluconate, 1% of thickening agent, 0.5% of vitamin C, 0.8% of vitamin E and the balance of water; the gelling agent is soluble starch, and the viscous agent is sodium carboxymethylcellulose; the preparation of the glue solution culture solution comprises the following steps: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding sodium carboxymethylcellulose, mixing uniformly, adding soluble starch for dissolving, then adding sucrose for dissolving, and finally adding vitamin C and vitamin E, mixing uniformly to obtain a glue solution culture solution;
(4) preparing a glue solution pollen culture solution: selecting flowers or buds with strong pollen activity and good development state on the male parent of the yolk fruit, brushing the pollen on a culture dish by using a brush pen, taking the 1/2 glue solution culture solution, adding 0.1 wt% of pollen into the glue solution culture solution, adding 0.01 wt% of boric acid, and mixing uniformly to prepare a glue solution pollen culture solution for later use;
(5) treatment of maternal stamens: injecting the liquid culture medium into the male core layer of the female parent by using an injector to wrap the anther by the liquid culture medium;
(6) and (3) cross pollination: when the pistil of female yolk fruit is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the pistil of female parent by using a dropper (as shown in figures 2 and 3).
The cross pollination method of the glue solution is simple in process and easy to operate, and can achieve the cross pollination rate of the yolk fruit of 50 flowers/hour, the fruit bearing rate of 41 percent and complete fruits.
Example 2
A glue solution cross pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent egg yolk fruit varieties with complementary advantages and similar flowering phases; the number of the female parent plants and the male parent plants is 10; the height of the yolk fruit plant is 275 cm;
(2) sleeving a pollen isolation net: selecting buds of which the stigma is to be exposed on side branches at the afternoon of 4:30-6:00, removing the buds which are not in the same period, using a pollen isolation mesh bag to sleeve the side branches and binding the side branches by using rubber bands or ropes, wherein the pollen isolation mesh is an ultra-dense nylon mesh, the aperture is 500 meshes, the diameter opening of the pollen isolation mesh bag sleeved on the side branches of the female parent is 50cm, and the bag depth is 100 cm; carrying out bagging operation on the male parent and the female parent;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing and placing in a refrigerating chamber with the temperature of 3 ℃, standing and refrigerating; the glue solution culture solution comprises 8% of sucrose, 2% of gelling agent, 0.5% of sodium gluconate, 0.5% of thickening agent, 0.3% of vitamin C, 0.5% of vitamin E and the balance of water; the gelling agent is soluble starch; the thickening agent is sodium carboxymethyl cellulose; the preparation of the glue solution culture solution comprises the following steps: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding sodium carboxymethylcellulose, mixing uniformly, adding soluble starch for dissolving, then adding sucrose for dissolving, and finally adding vitamin C and vitamin E, mixing uniformly to obtain a glue solution culture solution;
(4) preparing a glue solution pollen culture solution: selecting flowers or buds of the yolk fruit male parent with stronger pollen activity and good development state, brushing the pollen on a culture dish by using a brush, taking the 1/2 glue solution culture solution, adding 0.05 wt% of pollen into the glue solution culture solution, adding 0.005 wt% of boric acid, and mixing uniformly to prepare a glue solution pollen culture solution for later use;
(5) treatment of maternal stamens: injecting the liquid culture medium into the male core layer of the female parent by using an injector to wrap the anther by the liquid culture medium;
(6) and (3) cross pollination: when the female pistil of the egg yolk fruit female parent is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the female pistil of the female parent by using a dropper.
The cross pollination method of the glue solution is simple in process and easy to operate, and can achieve the cross pollination rate of the yolk fruit of 50 flowers/hour, the fruit bearing rate of 37 percent and complete fruits.
Example 3
A glue solution hybrid pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent yolk fruit varieties which have complementary advantages and similar flowering phases; the number of the female parent plants and the male parent plants is 8; the height of the yolk fruit plant is 230 cm;
(2) sleeving a pollen isolation net: selecting buds of which the stigma is to be exposed on the side branches at 4:30-6:00 in the afternoon, removing the buds which are not in the same period, and covering the side branches by using a shed type pollen isolation net which is a super-dense nylon net with the aperture of 1000 meshes; the operation of the pollen isolation net sleeved by the male parent is the same as that of the female parent;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing and placing in a refrigerating chamber with the temperature of 5 ℃, standing and refrigerating; the glue solution culture solution comprises 10% of cane sugar, 4% of gelling agent, 2% of sodium gluconate, 2% of thickening agent, 0.8% of vitamin C, 1% of vitamin E and the balance of water; the gel is agar; the thickening agent is hydroxypropyl methyl cellulose; the preparation of the glue solution culture solution comprises the following steps: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding hydroxypropyl methylcellulose, mixing uniformly, adding agar to dissolve, then adding sucrose to dissolve, and finally adding vitamin C and vitamin E, mixing uniformly to obtain a glue solution culture solution;
(4) preparing a liquid jelly pollen culture solution: selecting flowers or buds with strong pollen activity and good development state on the male parent of the yolk fruit, brushing the pollen on a culture dish by using a brush pen, taking the 1/2 glue solution culture solution, adding 0.2 weight percent of pollen into the glue solution culture solution, adding 0.02 weight percent of boric acid, and mixing uniformly to prepare a glue solution pollen culture solution for later use;
(5) treatment of maternal stamens: injecting the liquid jelly culture solution into the male core layer of female parent with injector to wrap the anther with the liquid jelly culture solution;
(6) and (3) cross pollination: when the female pistil of the egg yolk fruit female parent is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the female pistil of the female parent by using a dropper.
The glue solution hybrid pollination method has simple process and easy operation, and can realize the hybrid pollination rate of the yolk fruits of 50 flowers/hour, the fruit bearing rate of 39 percent and complete fruits.
Example 4
A glue solution hybrid pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting male parent and female parent wood milk fruit varieties which are strong and disease-free, have complementary advantages and have similar flowering phases; the number of the female parent plants and the male parent plants is 6; the height of the plant of the elaeagnus conferta roxb is 6 m;
(2) sleeving a pollen isolation net: selecting buds on lateral branches which are to be exposed out of stigma in 4:30-6:00 in the afternoon, removing the buds which are not in the same period, and wrapping dried flowers of the branches of the Moso milk fruits by using a sheet type pollen isolation net which is an ultra-dense nylon net with the aperture of 500 meshes; the male parent is sleeved with a pollen isolation net to operate the same as the female parent;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing and placing in a refrigerating chamber with the temperature of 3 ℃, standing and refrigerating; the glue solution culture solution comprises 8% of sucrose, 1% of gelling agent, 0.8% of sodium gluconate, 0.8% of thickening agent, 0.4% of vitamin C, 0.6% of vitamin E and the balance of water; the gel is agar; the thickening agent is sodium carboxymethyl cellulose; the preparation of the glue solution culture solution comprises the following steps: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding sodium carboxymethylcellulose, mixing uniformly, adding agar to dissolve, then adding sucrose to dissolve, and finally adding vitamin C and vitamin E, mixing uniformly to obtain a glue solution culture solution;
(4) preparing a glue solution pollen culture solution: selecting flowers or buds with stronger pollen activity and good development state on the milk fruit male parent, brushing the pollen on a culture dish by using a brush pen, taking the liquid culture medium, adding 0.08 wt% of pollen into the liquid culture medium, adding 0.008 wt% of boric acid, and mixing uniformly to prepare a liquid pollen culture medium for later use;
(5) and (3) cross pollination: when the pistil of the female parent of the elaeagnus conferta roxb is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the pistil head of the female parent by using a dropper (heterosexual plants of elaeagnus conferta roxb, male parent male flowers only need bagging).
The glue solution hybrid pollination method has simple process and easy operation, and can realize the hybrid pollination rate of the elaeagnus conferta roxb of 120 flowers/hour, the fruit bearing rate of 75 percent and complete fruits.
Example 5
A glue solution hybrid pollination method comprises the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent Annona squamosa L.varieties with complementary advantages and similar flowering phases; the number of the female parent plants and the male parent plants is 10; the height of the annona squamosa plant is 3 m;
(2) covering pollen to isolate the mesh bag: selecting buds to be cracked on side branches (as shown in figure 4) at 4:30-6:00 in the afternoon, removing the other buds which are not in the same period, using a pollen isolation mesh bag to sleeve the side branches and binding the side branches by using rubber bands or ropes, wherein the pollen isolation mesh is an ultra-dense nylon mesh with the aperture of 1000 meshes, the diameter opening of the pollen isolation mesh bag sleeved on the female parent branch dried flowers is 50cm, and the bag depth is 100 cm; carrying out bagging operation on the male parent and the female parent; the male parent is sleeved with a pollen isolation net to operate the same as the female parent; (ii) a
(3) Preparing a glue solution culture solution: preparing a glue solution culture solution, sealing and placing in a refrigerating chamber with the temperature of 3-5 ℃, standing and refrigerating; the glue solution culture solution comprises 10% of sucrose, 6% of gelling agent, 1.6% of sodium gluconate, 1.5% of thickening agent, 0.6% of vitamin C, 0.8% of vitamin E and the balance of water; the gelling agent is soluble starch; the thickening agent is hydroxypropyl methyl cellulose; the preparation of the glue solution culture solution comprises the following steps: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding hydroxypropyl methyl cellulose, uniformly mixing, adding soluble starch for dissolving, then adding cane sugar for dissolving, finally adding vitamin C and vitamin E, and uniformly mixing to obtain a glue solution culture solution;
(4) preparing a glue solution pollen culture solution: selecting flowers or buds with stronger pollen activity and good development state on the sweetsop male parent, brushing the pollen on a culture dish by using forceps and a brush, taking the 1/2 glue solution culture solution, adding 0.15 wt% of pollen into the glue solution culture solution, adding 0.015 wt% of boric acid, and mixing uniformly to prepare a glue solution pollen culture solution for later use;
(5) treatment of maternal stamens: injecting the liquid culture medium into the male core layer of female parent with syringe to wrap the anther with the liquid culture medium (as shown in FIG. 5);
(6) and (3) cross pollination: when the calyx of the female parent of Annona squamosa Linn is cracked, the prepared colloidal solution pollen culture solution is uniformly smeared on the head of the female parent female stamen by using a dropper (as shown in figure 6).
The glue solution hybrid pollination method has simple process and easy operation, and can realize the hybrid pollination rate of the sweetsop of 80 flowers/hour, the fruit bearing rate of 93 percent and complete fruits.
Comparative example 1
The difference between the comparative example 1 and the example 3 is that the yolk fruit female parent in the comparative example 1 does not use the colloidal solution culture solution and the colloidal solution pollen culture solution, when the female pistil of the yolk fruit female parent is exposed, the calyx is stripped by using tweezers, the male pistil is cut off, and then the male parent pollen is brushed on the female pistil of the female parent by using a brush.
The traditional method for carrying out hybrid pollination on the yolk fruits is complex in process and difficult to operate, the hybrid pollination rate of the yolk fruits is only 11 flowers/hour, the fruit bearing rate is only 12 percent, and the fruits are easy to damage.
Comparative example 2
Comparative example 2 is different from example 3 in that vitamin C and vitamin E were not added to the gum culture in comparative example 2.
The method for cross pollination of the yolk fruits is simple in process and easy to operate, and can realize that the cross pollination rate of the yolk fruits is 50 flowers/hour, but the fruit bearing rate is only 28 percent, and the fruits are complete.
The following table shows the corresponding comparative data for examples 1 to 5 and comparative examples 1 and 2:
by comparison, the cross pollination efficiency of the egg yolk fruit variety applied by the glue solution cross pollination method in the embodiments 1-3 of the invention can reach 50 flowers/hour, and the fruit bearing rate of the egg yolk fruit at the later stage is 37-41%. As can be seen from the comparison between the example 3 and the comparative examples 1 and 2, the comparative example 1 is a traditional hybridization pollination method, the operation is complicated, the fruits are easy to hurt, and only single combination hybridization can be carried out in the same bag, so that the pollination efficiency is low and the fruit bearing rate is not high; vitamin C and vitamin E were not added to the glue solution culture solution of comparative example 2, resulting in damage of photo-oxidation to the light and organs of the plants, and thus the fruiting rate was slightly lower than that of example 3. The complete technical scheme of the method of the invention is shown by comparing the embodiment 3 with the comparative examples 1 and 2 respectively, and the problems of labor and time waste, huge workload and low efficiency of manual emasculation (1) are solved; (2) flowers are easy to damage in the castration operation process, so that the normal development of fruits is influenced, and self-pollination is easy to cause if castration is not clean and timely, so that the hybridization rate is influenced; (3) the pollen is not easy to adhere when the pistil is fine and smooth, and the like.
From examples 4-5, it can be seen that the method of the present invention is applied to the cross pollination of Elaeagnus angustifolia and Annona squamosa varieties, indicating that the method of the present invention is suitable for the cross breeding of most plants. The method effectively reduces the working intensity of cross breeding, improves the working efficiency of cross breeding, and has great significance for accelerating the process and efficiency of cross breeding of special variety plants.
From the above, it can be seen that the present invention has a favorable effect of promoting the improvement of the efficiency of plant cross breeding.
The above description is intended to be illustrative of the preferred embodiment of the present invention and should not be taken as limiting the invention, but rather, the invention is intended to cover all modifications, equivalents, and alternatives falling within the spirit and scope of the present invention.
Claims (7)
1. A glue solution hybrid pollination method is characterized by comprising the following steps:
(1) preparation of male parent and female parent: selecting strong and disease-free male parent and female parent varieties which have complementary advantages and similar flowering phases; the variety is yolk fruit variety, wood milk fruit variety and sweetsop variety;
(2) sleeving a pollen isolation net: selecting male parent and female parent plants which contain buds to be released or have pistils to be exposed, and respectively covering the male parent and the female parent plants by using a pollen isolation net;
(3) preparing a glue solution culture solution: preparing a glue solution culture solution, sealing, standing and refrigerating; the glue solution culture solution comprises 8-10% of sucrose, 1-8% of gelling agent, 0.5-2% of sodium gluconate, 0.5-2% of thickening agent, 0.3-0.8% of vitamin C, 0.5-1% of vitamin E and the balance of water; the gel is 2-8% of soluble starch or 1-4% of agar; the thickening agent is sodium carboxymethyl cellulose or hydroxypropyl methyl cellulose;
(4) preparing a liquid jelly pollen culture solution: collecting pollen with strong activity on male parent, adding pollen and boric acid into the liquid jelly culture solution of 1/2, and mixing to obtain liquid jelly pollen culture solution; the weight percentage of the male parent pollen content in the liquid cement pollen culture solution is 0.05-0.2%, and the weight percentage of the boric acid content is 0.005-0.02%;
(5) treatment of maternal stamens: injecting the liquid culture medium into the male core layer of female parent to wrap the anther with the liquid culture medium;
(6) and (3) cross pollination: when the female parent flowers bloom or the pistil is exposed, the prepared colloidal solution pollen culture solution is uniformly smeared on the pistil heads.
2. The glue solution cross pollination method according to claim 1, wherein the glue solution culture solution is prepared by: taking the raw materials according to a certain proportion, dissolving sodium gluconate in water, adding a thickening agent, uniformly mixing, adding a gelling agent for dissolving, then adding cane sugar for dissolving, finally adding vitamin C and vitamin E, and uniformly mixing to obtain a glue solution culture solution.
3. The glue solution cross pollination method according to claim 1, wherein the glue solution pollen culture solution is prepared as it is.
4. The glue solution hybrid pollination method according to claim 1, wherein the refrigerating temperature of the glue solution culture solution in the step (3) is 3-5 ℃.
5. The glue solution hybrid pollination method according to claim 1, wherein the pollen collection in the step (4) is performed by brushing pollen on a culture dish by using a brush pen.
6. The glue solution cross pollination method as claimed in claim 1, wherein in the step (2), when the plant height is more than or equal to 150cm and less than or equal to 250cm, a shed pollen isolation net cover is used; when the plant height is more than 250cm or less than 150cm, the flower-bearing branches or the whole plant are sleeved by the bag-type pollen isolation mesh bag; when flowers are growing on the trunk of tall trees, the trunk flowers are wrapped with a sheet-type pollen isolation net.
7. The glue solution cross pollination method as claimed in claim 1 or 6, wherein the pollen isolation net in the step (2) is an ultra-dense nylon net with a pore size of 500-1000 meshes.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011188038.8A CN112493114B (en) | 2020-10-30 | 2020-10-30 | Glue solution hybrid pollination method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011188038.8A CN112493114B (en) | 2020-10-30 | 2020-10-30 | Glue solution hybrid pollination method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN112493114A CN112493114A (en) | 2021-03-16 |
| CN112493114B true CN112493114B (en) | 2022-08-19 |
Family
ID=74954543
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011188038.8A Active CN112493114B (en) | 2020-10-30 | 2020-10-30 | Glue solution hybrid pollination method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN112493114B (en) |
Families Citing this family (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114303932B (en) * | 2021-12-24 | 2022-10-28 | 中国热带农业科学院海口实验站 | Banana breeding method for one-time multi-comb pollination |
| CN114600768B (en) * | 2022-04-08 | 2023-03-31 | 开封市农林科学研究院 | Breeding method of high-resistance soybean |
| CN115568417B (en) * | 2022-10-24 | 2023-09-05 | 广西南亚热带农业科学研究所 | Liquid spray pollination method for macadimia nuts |
Family Cites Families (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP2001656B1 (en) * | 2006-04-06 | 2014-10-15 | 3D Systems Incorporated | KiT FOR THE PRODUCTION OF THREE-DIMENSIONAL OBJECTS BY USE OF ELECTROMAGNETIC RADIATION |
| CN101668855B (en) * | 2007-03-23 | 2013-07-03 | 丹尼斯科美国公司 | Enhanced amylase production by n-terminal addition to mature amylase protein |
| CN103461096B (en) * | 2013-08-20 | 2015-10-07 | 广西南亚热带农业科学研究所 | Cassava is broken flower bud hybridization pollination method manually |
| CN103670359A (en) * | 2013-12-10 | 2014-03-26 | 中国石油集团川庆钻探工程有限公司长庆井下技术作业公司 | Method for nondegradable gel breaking of vegetable gelatin fracturing fluid |
| WO2016034942A1 (en) * | 2014-09-02 | 2016-03-10 | Purecircle Usa Inc. | Stevia extracts enriched in rebaudioside d, e, n and/or o and process for the preparation thereof |
| WO2018204764A1 (en) * | 2017-05-05 | 2018-11-08 | Camp4 Therapeutics Corporation | Identification and targeted modulation of gene signaling networks |
| CN108617498B (en) * | 2018-05-02 | 2020-06-30 | 广西南亚热带农业科学研究所 | Efficient hybrid pollination method for pitaya |
-
2020
- 2020-10-30 CN CN202011188038.8A patent/CN112493114B/en active Active
Also Published As
| Publication number | Publication date |
|---|---|
| CN112493114A (en) | 2021-03-16 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN112493114B (en) | Glue solution hybrid pollination method | |
| Naik et al. | Nutrient-alginate encapsulation of in vitro nodal segments of pomegranate (Punica granatum L.) for germplasm distribution and exchange | |
| Kamenetsky et al. | Herbaceous peony (Paeonia): genetics, physiology and cut flower production | |
| CN105850725B (en) | A kind of method of dragon fruit pollination | |
| CN1124563A (en) | Methods for introducing a fertility restorer gene and for producing F1 producing F1 hybrid of brassica plants thereby | |
| Kishor et al. | Hybridization and in vitro culture of an orchid hybrid Ascocenda ‘Kangla’ | |
| Hall et al. | Isolation of a pure thornless loganberry by meristem tip culture | |
| Pradhan et al. | Efficient plant regeneration of Cymbidium aloifolium (L.) Sw., a threatened orchid of Nepal through artificial seed technology | |
| CN101263783B (en) | Method for using herbicide barban to fast breed aloe all-male plant strain | |
| CN105248266B (en) | A kind of upper implantation methods of dendrobium candidum tree | |
| Sakpere et al. | Effect of growth regulators and explant types on callus induction in Telfairia occidentalis Hook F | |
| Dag et al. | Effect of pollination method on fruit set and fruit characteristics in the vine cactus Selenicereus megalanthus (“yellow pitaya”) | |
| Marks | Rhododendron cuttings. I. Improved rooting following ‘rejuvenation’in vitro | |
| CN114097525B (en) | Method for cultivating pocket pumpkin-shaped seedless grape fruits | |
| US20190200553A1 (en) | Method for Producing Rice Haploid by Rice X Maize Hybridization | |
| CN107580965B (en) | Grape cultivation method | |
| CN110074111A (en) | A kind of inorganic agent and method improving millet Cross fertile rate | |
| CN114303933A (en) | Mechanical pollination method for konjac | |
| CN114831025A (en) | Rapid induction method of konjac polyploids | |
| Wetzstein et al. | Carya illinoensis (pecan) | |
| Muda et al. | Sugar palm (Arenga pinnata Wurmb Merr.): a review on plant tissue culture techniques for effective breeding | |
| KR101390396B1 (en) | Multiplication method of monocotyledon by removing stipes and by treating low temperature | |
| Abd Alhady | In Vitro Plant Regeneration from Alginate Encapsulated Shoot Tips of Casimairoa Edulis L. | |
| Blazich et al. | Propagation of fraser fir | |
| CN108703074B (en) | Temporary preservation and asexual propagation method of cabbage male sterile plant |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |