CN114391472A - Method for improving hybrid seed setting rate of pineapples - Google Patents
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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
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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Abstract
The invention provides a method for improving the seed setting rate of pineapple hybridization, which comprises the steps of collecting pollen after a parent strain is selected for flower forcing, spraying pollen treatment by using a pollen treatment agent containing calcium chloride, urea and ferric chloride to improve the cutinase activity in the pollen, promoting the pollen germination, improving the affinity of pineapple hybridization and further improving the insemination rate, then treating a maternal stigma by using a medicament containing cytokinin and citric acid to remove callose accumulation, promoting the pollen germination and pollen tube growth after pollination, and improving the seed setting rate. The method is simple and easy to master, does not need laboratory operation, can be completed in the field, can solve the problem of low seed setting rate of the pineapples due to hybridization affinity, and greatly improves the efficiency of pineapple hybridization breeding.
Description
Technical Field
The invention relates to the technical field of plant crossbreeding, in particular to a method for improving the hybrid seed setting rate of pineapples.
Background
The pineapple industry in China has the problems of single variety, degeneration of species and the like, and the improvement of the existing variety needs to be promoted by means of sexual hybridization and the like. However, the hybrid breeding work in China starts late, accumulates less and progresses slowly, and various problems exist in the hybrid breeding of pineapples, such as: the high self-incompatibility of the pineapples hinders the hybridization compatibility among varieties and lines, increases the difficulty of hybridization breeding, and also causes the phenomena of no or little seed formation of hybridized fruits in actual work, which seriously affect the smooth proceeding of the hybridization work, thereby hindering the hybridization breeding of the pineapples.
Chinese patent CN101861828A discloses a pineapple cross breeding method, which is characterized in that parents are reasonably configured, pollen which can be used for cross pollination is selected, pollination is carried out when small flower petals are opened, stigma is slightly seen and honeydew is secreted on the stigma, and the pineapple cross breeding method has low seed setting rate. In the hybridization breeding work, the common methods for overcoming incompatibility comprise the steps of treating stigma with medicaments, cutting stigma for pollination, grafting style, in vitro fertilization, hot water treatment, laser and high-voltage electrostatic field, pollen guide, ovary pollination and the like, the treatment effect is different due to different plant types, and the research report on pineapple hybridization incompatibility is less at present. Therefore, it is necessary to provide a method for improving the hybridization affinity between pineapple species and the seed set rate.
Disclosure of Invention
Therefore, the invention aims to provide a method for improving the hybrid seed setting rate of pineapples.
The technical scheme of the invention is realized as follows:
a method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting parent strains, and irrigating a flower accelerant to obtain hybrid parent strains;
b. pollen pretreatment: picking anther of male parent after flower bud blooms, taking out pollen from anther, spraying pollen with pollen treating agent, and air drying to obtain pollen; the pollen treating agent comprises calcium chloride, urea and ferric chloride;
c. treating stigma: smearing the maternal stigma with the medicament;
d. and (3) cross pollination: c, taking the pollen treated in the step b to carry out artificial pollination on the stigma treated in the step c, and closing the petals of the female parent after the pollination is finished;
e. seed taking: and cutting the hybridized small fruits and collecting seeds after the fig is fully yellow and ripe.
Further, the parent strain was of the varieties 'shenwan' (a. comosus cv. 'Shen Wan') and 'Yulinglong' (a. nanus cv. 'Yulinglong').
Furthermore, each 1L of the flower forcing agent contains 8-12 g of urea and 22-28 mL of ethephon, so that the flowering of parent plants is promoted, and the flowering period of the parent plants is unified.
Furthermore, the flower forcing agent is used by pouring from the top of the plant core, and the pouring amount is 40-50 mL/plant.
Further, the collection time of the anthers is 9-10 am, the buds burst in the period, and dew on the flowers is dried.
Furthermore, the pollen is taken out by tearing the anther and applying external force to the anther to naturally disperse the pollen in the anther so as to reduce the damage to the pollen.
Furthermore, the concentration of calcium chloride in the pollen treating agent is 0.2-0.5 wt%, the concentration of urea is 0.1-0.3 wt%, and the concentration of ferric chloride is 0.2-0.4 wt%.
Further, the use method of the pollen treating agent is that the pollen treating agent is applied to pollen in a spraying form; the application amount is 2-5 mL/g of pollen.
Further, the medicament comprises: cytokinin (CTK) and citric acid.
Furthermore, the 1L of the medicament contains 0.03-0.05 g of cytokinin and 0.1-0.2 g of citric acid.
Further, the temperature of pollen pretreatment and cross pollination is 20-30 ℃.
Further, the optimal temperature for pollen pretreatment and cross pollination is 28-30 ℃.
Compared with the prior art, the invention has the beneficial effects that:
the pollen treating agent containing calcium chloride, urea and ferric chloride is used for spraying and treating the pollen, so that the cutinase activity in the pollen can be improved, the pollen germination is promoted, the pineapple hybridization affinity is improved, and the insemination rate is further improved, which is shown in that the seed setting rate is increased.
The invention uses the medicament containing cytokinin and citric acid to treat the stigma, can remove callose accumulation without damaging the stigma, can promote pollen grain germination and pollen tube growth after pollination, and improves the seed setting rate.
The method is simple and easy to master, does not need laboratory operation, can be completed in the field, can solve the problem of low seed setting rate of the pineapples due to hybridization affinity, and greatly improves the efficiency of pineapple hybridization breeding.
Detailed Description
For clear and complete description of the technical solutions in the present invention, it is obvious that the inventor combines the embodiments to describe, but the following embodiments describe only some embodiments of the present invention, not all embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
The hybrid parent materials used in the experiment of the invention are 'Shenwan' (A. comosus cv. 'Shen Wan') and 'Yulinglong' (A. nanus cv. 'Yulinglong'), respectively;
the number of the hybridized flowers in each embodiment of the invention is 50;
example 1
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, pouring every 1L of flower forcing agent containing 10g of urea and 25mL of ethephon from the top of the plant heart, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 28 ℃, applying 3mL of the pollen treating agent to every 1g of the pollen, and drying in the air to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.3 wt%, the concentration of urea is 0.2 wt%, and the concentration of ferric chloride is 0.3 wt%;
c. treating stigma: smearing the stigma of the female parent with a preparation containing 0.04g cytokinin and 0.2g citric acid per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 2
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, and pouring flower forcing agent containing 8g of urea and 22mL of ethephon into every 1L of the top of the plant core, wherein the pouring amount is 50mL per plant, so as to obtain a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 30 ℃, applying 2mL of the pollen treating agent to every 1g of the pollen, and air-drying to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.5 wt%, the concentration of urea is 0.3 wt%, and the concentration of ferric chloride is 0.4 wt%;
c. treating stigma: smearing the stigma of the female parent with a preparation containing 0.03g cytokinin and 0.1g citric acid per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 30 ℃, lightly poking the dissilient female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the pollen to the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 3
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, pouring every 1L of flower forcing agent containing 10g of urea and 25mL of ethephon from the top of the plant heart, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 28 ℃, applying 3mL of the pollen treating agent to every 1g of the pollen, and drying in the air to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.3 wt%, the concentration of urea is 0.2 wt%, and the concentration of ferric chloride is 0.3 wt%;
c. treating stigma: smearing the stigma of the female parent with a medicament containing 0.04gABA and 0.2g 6-BA per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 4
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, pouring every 1L of flower forcing agent containing 10g of urea and 25mL of ethephon from the top of the plant heart, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 28 ℃, applying 3mL of the pollen treating agent to every 1g of the pollen, and drying in the air to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.1 wt%, the concentration of urea is 0.5 wt%, and the concentration of ferric chloride is 0.1 wt%;
c. treating stigma: smearing the stigma of the female parent with a preparation containing 0.04g cytokinin and 0.2g citric acid per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 5
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, pouring every 1L of flower forcing agent containing 10g of urea and 25mL of ethephon from the top of the plant core by taking the Gonwu pineapple as a female parent and the Yulinglong pineapple as a male parent, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 28 ℃, applying 3mL of the pollen treating agent to every 1g of the pollen, and drying in the air to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.3 wt%, and the concentration of urea in the pollen treating agent is 0.2 wt%;
c. treating stigma: smearing the stigma of the female parent with a preparation containing 0.04g cytokinin and 0.2g citric acid per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 6
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, pouring every 1L of flower forcing agent containing 10 urea and 25mL ethephon from the top of the plant heart, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen pretreatment: picking up anthers of male parents after flower buds bloom at 9-10 am, taking out pollen from the anthers, applying the pollen treating agent to the pollen in a spray mode at 28 ℃, applying 3mL of the pollen treating agent to every 1g of the pollen, and drying in the air to obtain the pollen; the concentration of calcium chloride in the pollen treating agent is 0.3 wt%, the concentration of urea is 0.2 wt%, and the concentration of ferric chloride is 0.3 wt%;
c. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
d. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Example 7
A method for improving the hybrid seed setting rate of pineapples comprises the following steps:
a. flower forcing: selecting a parent plant, taking 'Shenwan' pineapple as a female parent and 'Yulinglong' pineapple as a male parent, pouring every 1L of flower forcing agent containing 10g of urea and 25mL of ethephon from the top of the plant heart, wherein the pouring amount is 45mL per plant, and obtaining a hybrid parent plant;
b. pollen collection: picking anthers of the male parent after flower buds bloom at 9-10 am, and taking pollen out of the anthers;
c. treating stigma: smearing the stigma of the female parent with a preparation containing 0.04g cytokinin and 0.2g citric acid per 1L;
d. and (3) cross pollination: c, carrying out artificial pollination at 28 ℃, lightly poking the dissembled female parent petals with tweezers, clamping the male parent pollen processed in the step b with the tweezers, adhering the male parent pollen on the stigma processed in the step c, and lightly closing the petals;
e. seed taking: in the middle and last ten days of 6 months, when the fig is fully yellow and ripe, each hybrid fruit is cut off by a small knife, and seeds are collected.
Evaluation of hybridization Effect:
(1) the number of fruits with seeds, the number of seeds and the number of ovules in the above examples were counted, and the seed setting rate and the affinity index were calculated, and the results are shown in Table 1.
TABLE 1 hybridization affinity of pineapple treated by different methods
| Percentage of seed formation (%) | Affinity index | |
| Example 1 | 100 | 0.227 |
| Example 2 | 100 | 0.215 |
| Example 3 | 84.0 | 0.152 |
| Example 4 | 92.0 | 0.165 |
| Example 5 | 76.0 | 0.149 |
| Example 6 | 64.0 | 0.131 |
| Example 7 | 62.0 | 0.128 |
The result shows that the pollen treated by the pollen treating agent and the agent has high seed setting rate and affinity index for pollination, which are both obviously higher than the method of only using one of the pollen treating agent and the agent or using different components of the agent.
Test example 1
Hybridization combination: 'Yulinglong' × 'Shenwan';
the experiment was set up with 2 treatments, i.e., treatments with the treatment method of the invention and without the pollen treatment agent and agent of the invention applied, the cross pollination method being as in example 1 (except pollination temperature and hybridization combination);
the pollination temperature is respectively set to be 20 ℃, 28 ℃ and 35 ℃ (artificial climate box), each group combines 6 inflorescences, and each inflorescence has 13 florets.
TABLE 2 seed set percentage of Yulinglong ' x ' Shenwan ' at different temperatures
From table 2, it can be seen that the 'Yulinglong' x 'Shenwan' has the highest seed setting rate when the pollination temperature is 28 ℃, the affinity of the pollen treated by the method is improved, the seed setting rate is basically doubled at different hybridization temperatures, and the pollination seed setting rate is the highest at 28 ℃ and can reach 100%.
Test example 2
Hybridization combination: 'God gulf' × 'Yu exquisite', 'Yu exquisite' × 'God gulf';
hybridization method referring to example 1, the control was not treated with a pollen treatment agent spray.
TABLE 3 pineapple hybridization affinity case
As can be seen from Table 3, the pollen treated by the pollen treating agent and the agent of the invention is used for pollination, all pollinated small fruits bear seeds, the seed bearing rate reaches 100%, the affinity index is about 2 times of that of the control, the hybridization affinity is obviously improved, and the change of the parent and the female parent has no significant influence on the hybridization effect.
In conclusion, the method for the hybrid seed setting rate of the pineapples can effectively promote pollen germination and improve the affinity of pineapple hybridization. In addition, the preparation method is simple and easy to operate, and the cross breeding efficiency is high.
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, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (10)
1. A method for improving the hybrid seed setting rate of pineapples is characterized by comprising the following steps:
a. flower forcing: selecting parent strains, and irrigating a flower accelerant to obtain hybrid parent strains;
b. pollen pretreatment: picking anther of male parent after flower bud blooms, taking out pollen from anther, spraying pollen with pollen treating agent, and air drying to obtain pollen; the pollen treating agent comprises calcium chloride, urea and ferric chloride;
c. treating stigma: smearing the maternal stigma with the medicament;
d. and (3) cross pollination: c, taking the pollen treated in the step b to carry out artificial pollination on the stigma treated in the step c, and closing the petals of the female parent after the pollination is finished;
e. seed taking: when the fig is yellow and ripe, cutting the hybrid small fruits and collecting seeds.
2. The method for improving hybrid seed set percentage of pineapple as claimed in claim 1, wherein the parent strain is selected from the group consisting of God Bay and Yulinglong.
3. The method for improving hybrid seed setting rate of pineapple according to claim 1, wherein each 1L of said flower forcing agent contains 8-12 g of urea and 22-28 mL of ethephon; the flower forcing agent is poured from the top of the plant center, and the pouring amount is 40-50 mL/plant.
4. The method for increasing hybrid seed set rate of pineapple as claimed in claim 1, wherein said anther is collected at 9-10 am.
5. The method for improving hybrid seed setting rate of pineapple according to claim 1, wherein the concentration of calcium chloride, urea and ferric chloride in the pollen treating agent is 0.2-0.5 wt%, 0.1-0.3 wt% and 0.2-0.4 wt%.
6. The method for preparing the method for improving the hybrid seed setting rate of the pineapple as claimed in claim 5, wherein the pollen treating agent is applied by applying the pollen treating agent to pollen in a spray form; the application amount is 2-5 mL/g of pollen.
7. The method for preparing the method for improving the hybrid seed setting rate of the pineapple according to claim 1, wherein the medicament comprises: cytokinins and citric acid.
8. The method for preparing a method for improving hybrid seed set percentage of pineapple according to claim 7, wherein said agent contains 0.03-0.05 g cytokinin and 0.1-0.2 g citric acid per 1L.
9. The preparation method of the method for improving hybrid seed setting rate of pineapple according to claim 1, wherein the temperature of pollen pretreatment and hybrid pollination is 20-30 ℃.
10. The preparation method of the method for improving hybrid seed set percentage of pineapple according to claim 9, wherein the optimal temperature for pollen pretreatment and hybrid pollination is 28-30 ℃.
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| CN115362932A (en) * | 2022-09-14 | 2022-11-22 | 广西南亚热带农业科学研究所 | Pineapple film-covering moisture-proof hybridization method |
| CN116530261A (en) * | 2023-04-26 | 2023-08-04 | 广西南亚热带农业科学研究所 | Breeding method for comprehensively preventing and controlling pineapple hybrid seedling disease |
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| 张伟等: "‘神湾’菠萝与亲缘关系不同的父本的交配亲和性研究", 《中国园艺学会2019年学术年会暨成立90周年纪念大会论文摘要集》 * |
| 张伟等: "菠萝主栽品种间的杂交亲和力测定", 《中国园艺学会2018年学术年会论文摘要集》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN115362932A (en) * | 2022-09-14 | 2022-11-22 | 广西南亚热带农业科学研究所 | Pineapple film-covering moisture-proof hybridization method |
| CN116530261A (en) * | 2023-04-26 | 2023-08-04 | 广西南亚热带农业科学研究所 | Breeding method for comprehensively preventing and controlling pineapple hybrid seedling disease |
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| CN114391472B (en) | 2022-10-28 |
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