CN112740998B - Pollination method for pepper pollen in-vitro culture - Google Patents
Pollination method for pepper pollen in-vitro culture Download PDFInfo
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- CN112740998B CN112740998B CN202110117916.5A CN202110117916A CN112740998B CN 112740998 B CN112740998 B CN 112740998B CN 202110117916 A CN202110117916 A CN 202110117916A CN 112740998 B CN112740998 B CN 112740998B
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- 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
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Abstract
The invention discloses a pollination method for in-vitro culture of pepper pollen, belongs to the field of vegetable breeding, and particularly relates to the field of artificial pollination of vegetables, so as to solve the problems of large labor amount for collecting pollen, lack of professional technicians and high required scientific research expenditure in the conventional artificial pollination of pepper, and the method comprises the following steps: step 1, collecting and storing flower buds; step 2, stripping the anthers, removing impurities and collecting the anthers; step 3, placing the anthers in a constant-temperature illumination box for overnight culture; step 4, placing the anthers on a 55-65 mesh sieve for sieving, and collecting the anthers which are leaked below the sieve into a glass pollinator; and 5, manually emasculating the female parent pepper flowers, carrying out artificial pollination by using a glass pollinator, and marking the pollinated female flowers. The same materials are used in the full-bloom stage, 110-plus 140 plants can be pollinated by 1 day in the traditional method, and 220-plus 270 plants can be pollinated by 1 day in the method of the invention, so that the labor efficiency is obviously improved, and the labor cost is saved.
Description
Technical Field
The invention discloses a pollination method for in-vitro culture of pepper pollen, belongs to the technical field of vegetable breeding, and particularly relates to the technical field of artificial pollination of vegetables.
Background
The Capsici fructus is perennial herb of Magnoliaceae, Solanaceae, Capsicum, or limited number of years. Solitary flowers or fasciculate flowers, and drooping; the flowers are cup-shaped, and 5 teeth are not obvious; white or purple corolla, split oval; the anther is grayish purple. The fruit stalks are thicker and drooping; the fruits are long finger-shaped, horn-shaped, spherical, linear, etc., the top is new sharp and often bent, green when immature, red, orange or purplish red after mature, spicy. The seeds are flat kidney-shaped and light yellow.
When the hot pepper is pollinated artificially, the hot pepper which is already bloomed is prepared, pollen on hot pepper flowers is lightly scraped by a bamboo stick, meanwhile, the pollen is caught by paper under the hot pepper flowers, and the paper with the pollen is repeatedly smeared on pistils to help the hot pepper to pollinate.
Application No.: CN201310338345.3 discloses a method for spray pollination of pepper, which comprises the following steps: (1) collecting pepper male parent anther, airing to enable the pepper male parent anther to be in a powder-dispersed state, screening out pollen by using a 55-65-mesh sieve, and storing the pollen serving as a solid medium in a dry container; (2) storing pollen in an environment with humidity of 30-50% and temperature of 3-5 deg.C; (3) selecting a pepper female parent plant, and emasculating a flower bud one day before blooming; (4) artificial pollination is carried out on the same day of castration, and pollen with a solid medium prepared in advance is filled into a powder sprayer to be sprayed; when spraying, the powder spraying head of the powder sprayer aims at the emasculated stigma to spray the atomized pollen. The invention has the beneficial effects that: the fruit setting rate, pollination quality and pollination efficiency are improved, the labor cost is saved, the seed yield and quality are ensured, the pure pollen usage is reduced, the male parent planting proportion is reduced, the pollen collection labor is saved, and the seed reproduction cost is greatly reduced.
Above-mentioned patent is owing to need directly to gather open male flower and pollinate the female parent, and the pollen collection efficiency does not improve, and the recruitment volume is big.
Disclosure of Invention
The invention aims to: a pollination method for pepper pollen in-vitro culture aims at solving the problems that the existing artificial pollination of pepper is large in labor amount for pollen collection, professional technicians are lack of the labor amount, and the needed scientific research expenditure is high.
The technical scheme adopted by the invention is as follows:
a pollination method for isolated culture of pepper pollen comprises the following steps:
step 1, collecting and storing flower buds;
step 2, stripping the anthers, removing impurities and collecting the anthers;
step 3, placing the anthers in a constant-temperature illumination box for overnight culture for 12-18 hours;
step 4, placing the anthers on a 55-65 mesh sieve for sieving, and collecting the anthers which are leaked below the sieve into a glass pollinator;
and 5, manually emasculating the female parent pepper flowers, carrying out artificial pollination by using a glass pollinator, and marking the pollinated female flowers.
In the technical scheme of the application, the specific pollination method comprises the steps of collecting and storing buds; stripping anther, removing impurities and collecting anther; culturing anther under constant temperature, humidity and illumination; collecting pollen; because unopened flower buds are collected, after the pollen is cultured and matured, all pollen in the anthers can be collected, namely, the matured pollen is incubated in batches, so that sufficient pollen quantity for pollination is provided, the pollen culture medium is particularly suitable for materials with less male flowers of male parents, and is also very suitable for remote pollination; the same materials are used in the full-bloom stage, 110-plus 140 plants can be pollinated by 1 day in the traditional method, and 220-plus 270 plants can be pollinated by 1 day in the method of the invention, so that the labor efficiency is obviously improved, and the labor cost is saved.
Preferably, the environment temperature for collecting the buds in the step 1 is 25-28 ℃, the buds with intact developed petals and no cracking are collected, and no dew exists on the buds.
More preferably, the ambient temperature for collecting the flower buds in step 1 is 26.5 ℃.
Preferably, in the step 2, the calyx part is kneaded by a thumb and an index finger, so that anthers are integrally separated from the corolla and the pistil stigma; removing residual pistil stigma and impurities in the collected anther population.
Preferably, the collected anthers are placed in an ice box and taken back to the room to be cultured overnight in a constant-temperature light box with the temperature of 28-32 ℃ and the relative humidity of 40-50%, and the light is continuously irradiated with light intensity of 5000-.
More preferably, the temperature in the constant temperature light box is 30 ℃ and the relative humidity is 45%.
Preferably, the anthers are placed on a 60 mesh screen and sieved.
Preferably, the sieving specifically comprises the steps of lightly brushing off pollen by using a brush, leaking onto parchment paper or parchment paper laid below a sieve, collecting the anthers on the parchment paper or the parchment paper after all the anthers are sieved, disinfecting and drying the glass pollinator by using alcohol in advance, sealing the glass pollinator by using sponge, and placing the sealed glass pollinator into a vessel with a silica gel drying agent for storage.
Preferably, the lower side end of the glass pollinator is provided with a micropore, and the pistil stigma is inserted into the micropore in an aligning way by using a thumb and an index finger, so that the pistil stigma is fully adhered with anther.
More preferably, the pedicel on which the pistil stigma of the anther stick is located is marked.
Preferably, the refill material of the marking pen for marking is stamp pad oil mixed vegetable oil. The marked pedicel is not damaged, and the time is more durable and the color is not faded when the pedicel is marked in the field.
In the technical scheme of this application, in order to practice thrift the pollen, the micropore that pistil column cap inserted glass pollen pipe is very little, and this application micropore outside is provided with the little funnel of loudspeaker form, makes things convenient for the quick alignment micropore of column cap, improves pollination speed.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. in the invention, by adopting the method of artificially culturing pollen after the flower buds are separated, because unopened flower buds are collected, all pollen in anthers can be collected after the pollen is cultured to be mature, namely, the mature pollen is incubated in batches, and sufficient pollen quantity for pollination is provided;
2. in the invention, under the condition that the blooming number of the male parent can not meet pollination, in order to ensure pollen amount, flower buds with mature anthers are collected for artificial in-vitro culture, compared with the prior method of directly collecting open male flowers to pollinate the female parent, the invention obviously increases the workload of pollination days, shortens the pollination days and improves the pollination rate;
3. the method is particularly suitable for the materials with fewer male flowers of the male parent, and is also very suitable for pollination in different places, the weak male parent and the female parent pepper materials are not in the same test base, the pollen is directly peeled into the pollen tube and is stored in the drying box, and the pollen is pollinated in different places after being transported to different places, so that feasibility and convenience are brought to pollination in different places;
4. the workload and pollination rate of pollination days are improved. Under the condition that the artificial shortage of breeders exists, the workload of pollination days is improved, the total pollination days can be shortened, and the efficiency is improved;
5. due to the use of the glass pollinator, the uniform pollen adhesion of each pollinated pistil stigma can be ensured, and the pollination success rate reaches 99 percent;
6. in the present invention, pollen viability is ensured, and drying is required after the pollen is cultured in vitro.
Drawings
FIG. 1 is a drawing of the invention of flower bud collection;
FIG. 2 is a drawing showing the peeling of anthers according to the present invention;
FIG. 3 is a diagram of the present invention for removing impurities from anthers;
FIG. 4 is a drawing showing the peeling of the pollen of the present invention from the anther;
FIG. 5 is a diagram of a pollen-loaded glass pollinator according to the present invention;
FIG. 6 is a drawing of a pollinated and marked female stigma according to the present invention;
FIG. 7 is a diagram of pepper fruit growing after successful pollination according to the present invention;
FIG. 8 is a flow chart of a pollination method of the present invention.
Wherein the arrows in FIG. 6 indicate marked gynoecial stigma; in FIG. 7, the arrows indicate the growth of the pepper fruits after successful pollination.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1
As shown in FIGS. 1-8, a pollination method for in vitro culture of pepper pollen comprises the following steps:
step 1, collecting and storing flower buds; collecting buds at 25 deg.C without petal cracking, and collecting buds without dew;
step 2, stripping anthers, removing impurities, collecting anthers, pinching and rubbing calyx parts with a thumb and a forefinger to enable the anthers to be integrally separated from corolla and pistil stigma; removing residual pistil stigmas and impurities in the collected anther population;
step 3, placing the anthers in a constant-temperature illumination box for overnight culture for 18 hours, placing the collected anthers in an ice box and bringing the anthers back to the room to be cultured in the constant-temperature illumination box overnight, wherein the temperature in the constant-temperature illumination box is 28 ℃, the relative humidity is 45%, continuous illumination is carried out, and the illumination intensity is 5000 LUX;
step 4, placing the anthers on a 55-mesh sieve for sieving, wherein the anthers leak below the sieve and are collected into a glass pollinator, the sieving specifically comprises the steps of slightly brushing the pollen off by using a brush, leaking on parchment paper or parchment paper laid below the sieve, collecting the anthers on the parchment paper or parchment paper after all the anthers are sieved, disinfecting and drying the glass pollinator by using alcohol in advance, sealing the glass pollinator by using sponge, and placing the sealed glass pollinator in a vessel with a silica gel drying agent for storage;
and 5, manually castrating the flowers of the female parent capsicum, carrying out artificial pollination by using a glass pollinator, marking the pollinated female flowers, arranging micropores at the lower side end of the glass pollinator, aligning and inserting a pistil stigma into the micropores by using a thumb and a forefinger to ensure that the pistil stigma is fully adhered with anthers, marking the pedicels where the pistil stigma adhered with the anthers is positioned, and using a marking pen core material as stamp pad oil mixed vegetable oil for marking.
Example 2
As shown in fig. 1-7, on the basis of example 1, the ambient temperature of the flower buds collected in step 1 was 26.5 ℃; placing the anther in a constant-temperature illumination box for overnight culture for 14 hours in step 3, wherein the temperature in the constant-temperature box is 30 ℃, the relative humidity is 50%, and the continuous illumination is carried out with the illumination intensity of 8000 LUX; in step 4, the anthers were placed on a 60 mesh sieve and sieved, and the rest of the procedure was the same as in example 1.
Example 3
As shown in fig. 1-7, on the basis of example 1, the ambient temperature of the flower buds collected in step 1 was 28 ℃; placing the anther in a constant-temperature illumination box for overnight culture for 12 hours in step 3, wherein the temperature in the constant-temperature box is 32 ℃, the relative humidity is 40%, and continuous illumination is carried out, and the illumination intensity is 12000 LUX; in step 4, the anthers were placed on a 65 mesh sieve and sieved, and the rest of the procedure was the same as in example 1.
Test examples
Pollination tests were carried out in plastic greenhouses 1-1 and 1-2 of the experimental demonstration base of the Sichuan academy of agricultural sciences Pi. 1-1 shed is the pollination method, 1-2 sheds are the traditional pollination method as the contrast, and the parent materials sowed in 2 sheds are consistent. The test selects 5 different female parent pepper materials, each material is 100, pollen of a male parent material is subjected to isolated culture in a shed 1-1 at the 5-27-day full-bloom stage in 2020, flowers of each pepper are subjected to artificial emasculation in the 5-28-day month period, and artificial pollination and marking are carried out by using the glass pollinator assembled in the method. After 7 days, the pepper fruits are observed and counted. 1 skilled pollinator is dispatched in the shed for carrying out in-vitro culture on pollen of male parent material within 5 months and 27 days, and the working time is 1 hour; 2 skilled pollinators are dispatched for 5 months and 28 days, and pollination in the full-bloom stage is completed within 6 hours; meanwhile, pollination is carried out by a traditional method in a shed 1-2 in 28 days after 5 months, 4 skilled pollinators are dispatched, and pollination time is 7 hours, so that pollination in the full-bloom stage is completed. After 7 days, the pepper fruits are observed and counted. As a result, after 7 days, the pollination success rates of 5 different female parents in the 1-1 shed and the 1-2 sheds are consistent and indistinguishable. It can be seen that the present application is 2 times higher than the conventional method in saving labor cost.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents and improvements made within the spirit and principle of the present invention are intended to be included within the scope of the present invention.
Claims (7)
1. A pollination method for in vitro culture of pepper pollen is characterized by comprising the following steps:
step 1, collecting and storing buds, wherein the environment temperature of the collected buds is 25-28 ℃, the buds with intact developed petals and no cracking are collected, and no dew exists on the buds;
step 2, stripping the anther, removing impurities and collecting the anther, wherein the specific method comprises the following steps: pinching and rubbing calyx part with thumb and forefinger to make anther separate from corolla and pistil stigma; removing residual pistil stigmas and impurities in the collected anther population;
step 3, placing the anthers in a constant-temperature illumination box for overnight culture for 12-18 hours, placing the collected anthers in a box, bringing the collected anthers back to the room, and culturing the anthers in the constant-temperature illumination box overnight, wherein the temperature in the constant-temperature illumination box is 28-32 ℃, the relative humidity is 40-50%, and continuous illumination is carried out, and the illumination intensity is 5000-12000 LUX;
step 4, placing the anthers on a 55-65 mesh sieve for sieving, and collecting the anthers which are leaked below the sieve into a glass pollinator;
and 5, manually emasculating the female parent pepper flowers, carrying out artificial pollination by using a glass pollinator, and marking the pollinated female flowers.
2. The pollination method for the isolated culture of the pepper pollen as claimed in claim 1, wherein the temperature in the constant temperature illumination box is 30 ℃ and the relative humidity is 45%.
3. The pollination method of isolated culture of pepper pollen as claimed in claim 1, wherein the anther is placed on a 60 mesh sieve and sieved.
4. A pollination method for in-vitro culture of pepper pollen as claimed in claim 1 or 3, wherein the sieving is carried out by brushing pollen off gently with a brush, passing through parchment paper or parchment paper laid under the sieve, collecting anther on parchment paper or parchment paper after sieving, disinfecting and drying the glass pollinator with alcohol in advance, sealing the glass pollinator with sponge, and storing the sealed glass pollinator in a vessel with silica gel desiccant.
5. A pollination method for in vitro culture of pepper pollen as claimed in claim 1, wherein the lower side of the glass pollinator is provided with a micro hole, and the pistil stigma is inserted into the micro hole with the thumb and the index finger in alignment, so that the pistil stigma is fully adhered with anther.
6. A pollination method for in vitro culture of pepper pollen according to claim 5, wherein the pedicel on which pistil stigma of mucoid anther is located is marked.
7. A pollination method for in vitro culture of pepper pollen as claimed in claim 1 or 6, wherein the core material of the marking pen is stamp pad oil mixed rape oil.
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| CN103621397A (en) * | 2013-11-04 | 2014-03-12 | 饶燕 | Cultivation method of plantlet peppers |
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| CN103621397A (en) * | 2013-11-04 | 2014-03-12 | 饶燕 | Cultivation method of plantlet peppers |
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