CN112544435A - In-vitro hybridization method for gerbera jamesonii - Google Patents
In-vitro hybridization method for gerbera jamesonii Download PDFInfo
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- 238000000034 method Methods 0.000 title claims abstract description 27
- 238000009396 hybridization Methods 0.000 title claims abstract description 25
- 240000000208 Gerbera jamesonii Species 0.000 title claims abstract description 19
- 238000000338 in vitro Methods 0.000 title claims abstract description 12
- 230000010152 pollination Effects 0.000 claims abstract description 47
- 201000010099 disease Diseases 0.000 claims abstract description 10
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 claims abstract description 10
- 239000000463 material Substances 0.000 claims description 26
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 24
- 229920001131 Pulp (paper) Polymers 0.000 claims description 12
- 230000017260 vegetative to reproductive phase transition of meristem Effects 0.000 claims description 10
- 241000607479 Yersinia pestis Species 0.000 claims description 8
- 238000005286 illumination Methods 0.000 claims description 8
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 7
- 229930006000 Sucrose Natural products 0.000 claims description 7
- 229910000402 monopotassium phosphate Inorganic materials 0.000 claims description 7
- 235000019796 monopotassium phosphate Nutrition 0.000 claims description 7
- PJNZPQUBCPKICU-UHFFFAOYSA-N phosphoric acid;potassium Chemical compound [K].OP(O)(O)=O PJNZPQUBCPKICU-UHFFFAOYSA-N 0.000 claims description 7
- 239000005720 sucrose Substances 0.000 claims description 7
- 241000196324 Embryophyta Species 0.000 claims description 6
- 241000238631 Hexapoda Species 0.000 claims description 6
- 239000007788 liquid Substances 0.000 claims description 6
- 239000003755 preservative agent Substances 0.000 claims description 6
- 230000002335 preservative effect Effects 0.000 claims description 6
- GTOQWWQKBBZILU-UHFFFAOYSA-N 2-hydroxypropane-1,2,3-tricarboxylic acid;quinolin-8-ol Chemical compound C1=CN=C2C(O)=CC=CC2=C1.OC(=O)CC(O)(C(O)=O)CC(O)=O GTOQWWQKBBZILU-UHFFFAOYSA-N 0.000 claims description 5
- 238000007598 dipping method Methods 0.000 claims description 3
- 241000735332 Gerbera Species 0.000 claims description 2
- 235000016709 nutrition Nutrition 0.000 claims description 2
- 230000035764 nutrition Effects 0.000 claims description 2
- 238000012258 culturing Methods 0.000 claims 1
- 238000009395 breeding Methods 0.000 abstract description 13
- 230000001488 breeding effect Effects 0.000 abstract description 13
- 230000002411 adverse Effects 0.000 abstract 1
- 239000003761 preservation solution Substances 0.000 description 9
- 238000009402 cross-breeding Methods 0.000 description 7
- 239000000243 solution Substances 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- KRKNYBCHXYNGOX-UHFFFAOYSA-K Citrate Chemical compound [O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O KRKNYBCHXYNGOX-UHFFFAOYSA-K 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000007613 environmental effect Effects 0.000 description 2
- 238000003306 harvesting Methods 0.000 description 2
- 241000208838 Asteraceae Species 0.000 description 1
- 241000221662 Sclerotinia Species 0.000 description 1
- 238000007596 consolidation process Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000010154 cross-pollination Effects 0.000 description 1
- 238000005034 decoration Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 230000018109 developmental process Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 238000010353 genetic engineering Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 239000000543 intermediate Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000003147 molecular marker Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
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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
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N3/00—Preservation of plants or parts thereof, e.g. inhibiting evaporation, improvement of the appearance of leaves or protection against physical influences such as UV radiation using chemical compositions; Grafting wax
- A01N3/02—Keeping cut flowers fresh chemically
Abstract
The invention discloses an in vitro hybridization method of gerbera jamesonii, belonging to the technical field of gerbera jamesonii hybridization breeding. The invention obtains hybrid seeds by collecting male parent pollen and carrying out excised vase pollination on the cut gerbera jamesonii flowers. The method provides a new way for breeding work, overcomes the adverse factors of high temperature, high humidity, diseases and the like which are easy to appear in the gerbera jamesonii hybridization process, and obviously improves the success rate of gerbera jamesonii hybridization.
Description
Technical Field
The invention belongs to the technical field of gerbera jamesonii crossbreeding, and particularly relates to a method for obtaining hybrid seeds by using excised pollination of gerbera jamesonii cut flowers.
Background
The gerbera jamesonii is perennial herb flower of gerbera of Compositae, has large flower, rich flower color, long fresh-keeping period, annual flower, long-distance transportation resistance and strong decoration, is an ideal material for making gift bouquets, flower baskets and artistic flower arrangement, is one of five fresh cut flowers in the world, and has wide market prospect.
African daisy native south Africa, China is not the native place of African daisy, so breeding mainly depends on foreign mature varieties for breeding. With the rapid development of modern biotechnology, a large number of advanced breeding techniques are developed and applied, including mutation breeding, ploidy breeding, molecular marker assisted breeding, genetic engineering breeding and the like, but the traditional cross breeding is still the most important and most common breeding means.
At present, in the cross breeding process of the gerbera jamesonii, the success rate of cross pollination is greatly influenced by environmental conditions such as temperature and humidity. After hybridization pollination, if the air humidity is high, inflorescences are easy to mildew and rot, so that the hybridization failure is caused; the phenomenon of low or even non-consolidation of the hybridization is easy to occur under the high-temperature and drought climatic conditions. In most areas of China, the greenhouse humidity is high due to the fact that the greenhouse humidity is high in spring rain and humidity, high-temperature drought in summer and low-temperature greenhouse humidity in winter need of film covering and heat preservation, selective cross breeding time is very limited, and breeding efficiency is reduced.
Disclosure of Invention
The invention provides a method for obtaining hybrid seeds by in vitro pollination of cut flowers of gerbera jamesonii, which aims to solve the problems.
The invention adopts the following technical scheme:
a method for obtaining hybrid seeds by in vitro pollination of cut gerbera jamesonii flowers comprises the following steps of collecting male parent pollen and carrying out in vitro vase pollination on the cut gerbera jamesonii flowers, and specifically comprises the following steps:
1) selecting male parent materials: the flower branch with regular flower type, no disease and pest damage and pollen scattering of anther in 2-4 rounds of inflorescence is selected as the male parent of hybridization.
2) Pollen collection: when the pollen of the male parent flower branches is scattered, the flower branches are folded down, the pollen is shaken into a culture dish with the diameter of 7cm or 9cm, and the dish cover is closed and taken back to the room for pollination.
3) Selecting a female parent material: selecting flowering branches with the length of the flower stalk of 50-70cm, the thickness of the flower stalk of 0.6-1.0cm, flat tongue-shaped flowers of the outer round, no pollen emission of anthers of the first round, regular flower types and no diseases or insect pests as female parent materials.
4) Treating a mother material: the outer wheel tongue-shaped petals are firstly cut off by scissors, and the allowance of 0.3-0.8cm is left, so that the stigma is prevented from being injured. Then, a sharp knife is used for obliquely cutting off the part 2-3cm away from the base of the pedicel at the angle of 30-45 ℃, the cut part is put into a vase with preservation solution for culture, the vase specification is that the vase is 30cm high and 10cm in diameter, 8-12 female parent flowering branches are placed on each vase, the water level height of the preservation solution is 5-8cm, and the preservation solution comprises 20g/L sucrose, 200 mg/L8-hydroxyquinoline citrate and 60mg/L potassium dihydrogen phosphate.
5) Pollination: the pollen of the male parent in the culture dish is dipped by a brush pen and smeared on the petal stigmas of the outer wheel of the female parent for 2-3 rounds, and the pollen is not suitable to be fully pollinated to ensure that the nutrition supply is sufficient and the obtained seeds are full. And (5) sleeving a sulfuric acid paper bag after pollination is finished, and registering and listing.
6) And (3) management after pollination: after pollination, the temperature of the female parent material is controlled to be 20-25 ℃, the humidity is controlled to be 45-75%, meanwhile, the illumination intensity is 500-2000lux, and the illumination time is 10-14 h. And 3-5d after pollination, taking down the sulfuric acid paper bag. Changing the fresh-keeping liquid every 2-4 days after pollination, cutting off the base part of the flower stalk by a sharp knife at the angle of 30-45 ℃ for 2-5cm while changing the fresh-keeping liquid, wherein the formula of the fresh-keeping liquid is 20g/L sucrose, 200 mg/L8-hydroxyquinoline citrate and 60mg/L potassium dihydrogen phosphate.
7) Seed collection: and (4) 18-30d after pollination, the disk-shaped flowers in the middle of the inflorescence are completely opened, and the seeds can be harvested when the seeds are mature.
Compared with the conventional hybridization method, the method has the advantages that:
first, the method of the invention has high hybridization success rate, compared with the conventional method, the method of the invention has easier and more accurate temperature and humidity control, and can avoid the problems that the hybridization success rate is influenced by the mildewing of inflorescences, difficult fructification of hybridization and the like after hybridization. Meanwhile, the invention adopts the mode of cut flower in vitro pollination, so that the death and the failure of hybridization of the parent plant caused by the damage of diseases such as root rot, sclerotinia, southern blight and the like to the hybrid parent plant after hybridization can be avoided.
Secondly, the method is not limited by seasons and external environmental conditions, and can carry out cross breeding all the year round, thereby remarkably accelerating the breeding process.
Third, the method of the present invention is more efficient in cross breeding of a small amount of valuable materials or breeding intermediates than conventional cross breeding methods.
Drawings
FIG. 1 shows that conventional hybridization methods are prone to mildew of inflorescences;
FIG. 2 shows that conventional hybridization methods are prone to decay of inflorescences and mildew of seeds;
FIG. 3 is a schematic diagram of a cut flower of African daisy in vitro pollination hybridization method;
FIG. 4 in vitro pollination of normal inflorescences;
FIG. 5 in vitro pollination of mature seeds;
FIG. 6 in vitro pollination of seeds for normal germination.
The specific implementation mode is as follows:
example 1
A hybridization method of African daisy comprises the following steps:
(1) parent material selection: in 3-5 months, the flower branches with regular flower type, no plant diseases and insect pests and 2-3 rounds of anthers in the inflorescence begin to shed pollen are selected as male parents for hybridization. Selecting the flower branch with the length of 60-70cm, the thickness of 0.6-0.8cm, the flat and flat outer wheel lingulate flower, the first round of anther without powder, regular flower type and no disease or pest as the female parent material. .
(2) Pollen collection: when the pollen of the male parent flower branches is scattered, the flower branches are folded down, the pollen is shaken into a culture dish with the diameter of 7cm, and the dish cover is closed and is taken back to the room for pollination.
(3) Treating a mother material: taking off female parent flower branch, taking back to indoor, cutting off foreign wheel tongue shape petal with scissors, and leaving 0.3-0.8cm of margin to avoid hurting stigma. Meanwhile, a sharp knife is used for obliquely cutting off the length of 2-3cm of the base part of the pedicel at the angle of 30-45 ℃, the pedicel is placed into a vase with the caliber of 10cm and the height of 30cm for culture, 8 flowering branches are placed on each vase, the water level of the preservative solution is 5cm, and the components of the preservative solution are 20g/L of sucrose, 200mg/L of 8-hydroxyquinoline citrate and 60mg/L of potassium dihydrogen phosphate.
(4) Pollination: dipping the pollen of the male parent in the culture dish by using a writing brush, smearing the pollen on the petal column of the outer wheel of the female parent for 2-3 times. And (5) sleeving a sulfuric acid paper bag after pollination is finished, and registering and listing.
(5) And (3) management after pollination: the mother plant material is moved into an artificial climate chamber for management, and the flowering branches are placed against a wall. The temperature of the artificial climate chamber is controlled to be 23-25 ℃, the humidity is 45-55%, the illumination time is 10h, and the illumination intensity is 1500-. And 3d after pollination, taking down the sulfuric acid paper bag, simultaneously changing the preservation solution every 2-3d, cutting off 3-5cm of the base part of the pedicel at the angle of 30-45 ℃ while changing the preservation solution, wherein the components of the preservation solution are sucrose with the final concentration of 20g/L, 8-hydroxyquinoline citrate with the final concentration of 200mg/L and potassium dihydrogen phosphate with the final concentration of 60 mg/L.
(6) Seed collection: and (4) 18-23d after pollination, the disk-shaped flowers in the middle of the inflorescence are completely opened, and the seeds can be harvested when the seeds are mature.
Example 2
(1) Parent material selection: in 7-9 months, the flower branches with regular flower type, no plant diseases and insect pests and 3-4 rounds of anthers in the inflorescence begin to shed pollen are selected as male parents for hybridization. Selecting flowering branches with the length of the flower stalk of 50-60cm, the thickness of the flower stalk of 0.7-0.9cm, flat tongue-shaped flowers of the outer round, no pollen emission of anthers of the first round, regular flower types and no diseases or insect pests as female parent materials.
(2) Pollen collection: when the pollen of the male parent flower branches is scattered, the flower branches are folded down, the pollen is shaken into a culture dish with the diameter of 9cm, and the dish cover is closed and is taken back to the room for pollination.
(3) Treating a mother material: taking off female parent flower branch, taking back to indoor, cutting off foreign wheel tongue shape petal with scissors, and leaving 0.3-0.8cm of margin to avoid hurting stigma. Meanwhile, a sharp knife is used for obliquely cutting off the length of 2-3cm of the base part of the pedicel at the angle of 30-45 ℃, the pedicel is placed into a vase with the caliber of 10cm and the height of 30cm for culture, 12 flowering branches are placed in each vase, the water level of the preservative solution is 8cm, and the components of the preservative solution are 20g/L of sucrose, 200mg/L of 8-hydroxyquinoline citrate and 60mg/L of potassium dihydrogen phosphate.
(4) Pollination: dipping the pollen of the male parent in the culture dish by using a writing brush, smearing the pollen on the petal column of the outer wheel of the female parent for 2-3 times. And (5) sleeving a sulfuric acid paper bag after pollination is finished, and registering and listing.
(5) And (3) management after pollination: the mother material is moved into an artificial climate chamber for management, and the flowering branch is close to the wall. The temperature of the artificial climate chamber is controlled to be 20-22 ℃, the humidity is 65-75%, the illumination time is 14h, and the illumination intensity is 500-. And 5d after pollination, taking down the sulfuric acid paper bag, simultaneously changing the preservation solution every 3-4d, cutting off 2-4cm of the base part of the pedicel at the angle of 30-45 ℃ while changing the preservation solution, wherein the components of the preservation solution comprise 20g/L of sucrose, 200mg/L of 8-hydroxyquinoline citrate and 60mg/L of potassium dihydrogen phosphate.
(6) Seed collection: and (5) after pollination, the disk-shaped flowers in the middle of the inflorescence are completely opened 23-30 days, and the seeds can be harvested when the seeds are mature.
Comparative example 1
Cutting the female parent's lingulate petals to short (about 0.3-0.8cm away from stigma) in 3-5 months, smearing the pollen of male parent on the stigma of female parent, registering and tagging, covering with sulfuric acid paper bag, taking off sulfuric acid paper bag one week after pollination, and harvesting after seeds are mature.
Comparative example 2
Cutting the female parent's lingulate petals to short (about 0.3-0.8cm away from stigma) in 7-9 months, smearing the pollen of the male parent on the stigma of the female parent, registering and tagging, sleeving a sulfuric acid paper bag, taking down the sulfuric acid paper bag one week after pollination, and harvesting after the seeds are mature.
The number of pollinated flowers, the number of seeds bearing seeds and the success rate of hybridization by the method of the above example are shown in Table 1.
TABLE 1
The above description is only a preferred embodiment of the present invention, and all equivalent changes and modifications made in accordance with the claims of the present invention should be covered by the present invention.
Claims (3)
1. A method for obtaining hybrid seeds by in vitro pollination of cut flowers of gerbera jamesonii, which is characterized by comprising the following steps:
1) selecting male parent materials; 2) collecting pollen; 3) selecting a female parent material; 4) treating the parent material; 5) pollinating; 6) managing after pollination; 7) seed collection;
step 3) selecting flowering branches with the flower stalk length of 50-70cm and the flower stalk thickness of 0.6-1.0cm as the female parent material;
step 4) the mother material treatment method comprises: firstly, cutting off the tongue-shaped petals of the outer wheel by using scissors, then cutting off the base part of the flower stalk, putting the flower pot into a vase with a fresh-keeping solution for culturing, wherein the specification of the vase is that the height of the vase is 30cm, the diameter is 10cm, and 8-12 female parent flower branches are placed in each vase;
step 5), smearing the pollination stigma for 2-3 rounds;
and 6) controlling the temperature of the female parent material to be 20-25 ℃ and the humidity to be 45-75% in the management step after pollination.
2. The method for obtaining hybrid seeds by the excised pollination of the cut gerbera flowers according to claim 1, which comprises the following steps:
selecting male parent materials: selecting flowering branches with regular flower type, no plant diseases and insect pests and pollen scattering of anthers of 2-4 rounds in inflorescence as male parents for hybridization;
pollen collection: when the pollen of the male parent flower branches is scattered, the flower branches are folded down, the pollen is shaken into a culture dish with the diameter of 7cm or 9cm, and the dish cover is closed and is taken back to the room for pollination;
selecting a female parent material: selecting flowering branches with the length of the flower stalk of 50-70cm, the thickness of the flower stalk of 0.6-1.0cm, flat tongue-shaped flowers of the outer wheel, no pollen emission of anthers of the first round, regular flower types and no diseases or insect pests as a female parent material;
treating a mother material: firstly, cutting off the tongue-shaped petals of the outer wheel by using scissors, and leaving a margin of 0.3-0.8cm to avoid hurting the column head; then, a sharp knife is used for obliquely cutting off the part 2-3cm away from the base of the pedicel at the angle of 30-45 ℃, the cut flower is put into a vase with fresh-keeping liquid for culture, the vase specification is that the height of the vase is 30cm, the diameter is 10cm, and each vase is provided with 8-12 female parent flower branches;
5) pollination: dipping the pollen of the male parent in the culture dish by using a writing brush, smearing the pollen to the petal column heads of the outer wheel of the female parent for 2-3 rounds, and not being suitable for full pollination so as to ensure sufficient nutrition supply and full obtained seeds; sheathing a sulfuric acid paper bag after pollination, and registering and listing;
6) and (3) management after pollination: after pollination, the temperature of the female parent material is controlled at 20-25 ℃, the humidity is 45-75%, meanwhile, the illumination intensity is 500-2000lux, and the illumination time is 10-14 h; 3-5 days after pollination, taking down the sulfuric acid paper bag; changing the fresh-keeping liquid every 2-4 days after pollination, and cutting off the base part of the flower stalk by 2-5cm at an angle of 30-45 ℃ by using a sharp knife while changing the fresh-keeping liquid;
7) seed collection: and (4) 18-30 days after pollination, the disk-shaped flowers in the middle of the inflorescence are completely opened, and the seeds are collected when the seeds are mature.
3. The method according to claim 1 or 2, wherein the height of the preservative solution is 5-8cm, and the components of the preservative solution comprise sucrose with a final concentration of 20g/L, 8-hydroxyquinoline citrate with a final concentration of 200mg/L and potassium dihydrogen phosphate with a final concentration of 60 mg/L.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112369328A (en) * | 2020-11-15 | 2021-02-19 | 云南省农业科学院花卉研究所 | Rapid cultivation method of gerbera jamesonii excellent strain |
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Cited By (2)
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