CN107667861B - Grafting method of peanut tissue culture seedlings - Google Patents
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- CN107667861B CN107667861B CN201710995683.2A CN201710995683A CN107667861B CN 107667861 B CN107667861 B CN 107667861B CN 201710995683 A CN201710995683 A CN 201710995683A CN 107667861 B CN107667861 B CN 107667861B
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Abstract
The invention discloses a grafting method of peanut tissue culture seedlings, which comprises the steps of culturing black peanut seeds under a dark culture condition, selecting seedlings with hypocotyl length of 6-8cm as stocks when two true leaves of peanut seedlings are unfolded, cutting off cotyledons and parts above the cotyledons of the seedlings, and longitudinally cutting the middle of the hypocotyl into a notch; and taking the sterile tissue culture seedlings as scions, grafting and transplanting to a field. The method utilizes the dark condition to culture the seedlings, can ensure that the hypocotyls of the seedlings grow quickly, starts grafting when the length of the hypocotyls is 6-8cm, can avoid the problems that the hypocotyls of stock materials cultured by common illumination are short (1-2cm) and are not easy to operate during grafting, and directly improves the grafting efficiency and the grafting survival rate. Compared with the method that only more than ten plants can be grafted in 1h when the hypocotyl is short, the method can graft more than 30 plants in 1h when the hypocotyl is long, and the working efficiency is improved by more than one time.
Description
Technical Field
The invention belongs to the technical field of agricultural planting biology, and particularly relates to a grafting method of peanut tissue culture seedlings.
Background
Peanuts are important oil and economic crops, occupy an important position in agricultural production, the peanut planting area of China is gradually enlarged in recent years, but the genetic basis of the currently popularized peanut cultivar is narrow, and the disease resistance and stress resistance are poor, so that the yield and the quality of the peanuts are further improved. With the rapid development of biotechnology, transgenic and in vitro mutagenesis technologies are valued by researchers; with the gradual improvement of the distant hybridization technology of peanuts, the selection of the wild species with excellent properties such as disease resistance, stress tolerance and the like for hybridization with the cultivated species becomes an important way for breeding new disease-resistant and stress tolerance excellent varieties, but most of distant hybridization shows incompatibility of hybridization and abortion of the hybrids, embryo rescue needs to be carried out through tissue culture, the hybrid F1 is obtained, and after chromosome doubling treatment, the seedlings are transplanted to the field, but the roots of the wild species and the progeny materials are weak, and some materials are difficult to root, so that the transplanting survival rate of the tissue culture seedlings is extremely low, the growth vigor after the transplantation is poor, and the smooth progress of the breeding work is seriously influenced.
The grafting technology is widely applied to horticultural crops, the application reports of peanuts are few, and with the continuous development of the grafting technology, cotton, soybeans, mung beans, sweet potatoes and the like obtain new germplasm through distant grafting, so that the traditional propagation significance of the new germplasm is broken through at present, and the new germplasm is widely applied to the aspects of cultivation production, breeding practice, basic theoretical research and the like of plants. In the aspect of peanut grafting, researches report that 10-14d peanut seedlings are used as stocks, and different grafting methods (cleft grafting and conjugal grafting) and different grafting parts (epicotyl and hypocotyl) are adopted. However, after the epicotyl is grafted and transplanted into the field, the cotyledonary node has a bud primordium, and a lateral bud can grow after grafting, so that axillary buds need to be removed frequently, the field workload is increased, and the growth and fructification of the scion are also influenced. In the conventional hypocotyl grafting, because the hypocotyl is short, the hypocotyl is almost in soil, the operation is not good during the grafting, and the grafting efficiency and the grafting survival rate are directly influenced. There is also a report on adopting an aseptic grafting method, using aseptic seedlings with the age of 12-15d as stocks, using peanut tissue culture seedlings as scions, grafting on hypocotyls, grafting under aseptic condition, then culturing in an aseptic culture medium to heal wounds, taking out the grafted seedlings after survival, washing the culture medium on the root system of the grafted plants with water, then transplanting into a seedling culture medium, domesticating indoors for 3 weeks, and transplanting into the field after survival. Because the sterile grafting is adopted, the seeds need to be disinfected and inoculated in a culture medium, the whole growth process is ensured to be sterile, and certain obstacles are generated in the operation during the grafting.
Disclosure of Invention
Aiming at the problems and the defects existing in peanut grafting, the invention provides the grafting method of the peanut tissue culture seedlings, which can improve the transplanting survival rate, simplify the grafting procedure, shorten the growth period and enable the transplanted seedlings to grow robustly in the field.
In order to achieve the purpose, the invention adopts the technical scheme that:
a grafting method of peanut tissue culture seedling comprises culturing black peanut seed in dark culture condition, selecting seedling with hypocotyl length of 6-8cm as stock when two true leaves of peanut seedling are unfolded, cutting off cotyledon and part above the seedling, and longitudinally cutting hypocotyl middle into cut; taking the aseptic tissue culture seedling as a scion, cutting the lower end of the scion into a V-shaped wound corresponding to the cut, and keeping the cut flat; inserting the lower end of the scion into the cut of the stock, and sealing a film to form a grafted seedling; and (4) placing the grafted seedling indoors, and transplanting the grafted seedling into a field after new leaves grow out.
The grafting method of the peanut tissue culture seedling comprises the following specific steps:
(1) culturing black peanut seeds under a dark culture condition, selecting seedlings with hypocotyl length of 6-8cm as stocks when two true leaves of peanut seedlings are unfolded, cutting off cotyledons and parts above the cotyledons of the peanut seedlings, longitudinally cutting the middle of the hypocotyls into cuts with the depth of 0.8-1 cm;
(2) taking the sterile tissue culture seedlings as scions, cutting the lower ends of the scions into V-shaped wounds corresponding to the cuts, and keeping the cuts flat; inserting the lower end of the scion into the cut of the stock, and tightly winding the scion with a Parafilm sealing film to enable the stock and the scion to be tightly combined to form a grafted seedling;
(3) watering, namely placing the grafted seedlings indoors to avoid direct sunlight and covering the grafted seedlings with a plastic film; removing the film after new leaves grow out, and gradually culturing with light;
(4) transplanting the grafted seedlings with good growth vigor to a field in a cloudy day or at the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; and after the transplanted seedlings completely survive, removing the sunshade net, and performing normal field management.
The black peanut variety is Yuhua 0215.
The method for culturing the black peanut seeds in the dark comprises the following steps: soaking black peanut seeds overnight for accelerating germination, then planting the seeds in a nutrition pot containing sterilized vermiculite, placing the seeds in the dark at 27 ℃ for culturing until two true leaves are unfolded, and selecting seedlings with the hypocotyl length of 6-8cm as stocks.
The tissue culture seedling is a wild peanut seed, an amphidiploid or a transgenic plant.
The height of the tissue culture seedling is more than 2 cm.
The length of the two inclined planes of the V-shaped wound is 0.8-1 cm.
The indoor environment is at 27 ℃ and 50% humidity.
And (4) after new leaves grow out from the grafted seedlings in the step (3), placing the grafted seedlings in the sun for 1 hour in the first day, increasing the illumination time day by 0.5-1.5 hour every day, and transplanting the grafted seedlings to a field after 7 days.
The invention has the beneficial effects that:
1. important peanut wild species materials and transgenic materials need to be propagated and transplanted into fields through tissue culture, but the root systems of the wild species and progeny materials are weak, so that the transplanting survival rate of tissue culture seedlings is extremely low. The invention solves the problems that the rooting of the wild species and the offspring materials is difficult, the transplanting survival rate is low, the growth vigor of the tissue culture seedlings after being transplanted is weak, and the like, saves the step of rooting, shortens the growth period, and ensures that the root system of the stock is vigorous, the growth vigor in the transplanting field is strong and the bearing performance is good.
2. According to the method, the black peanut variety is selected as the stock, grafted, survived and transplanted to the field, even if axillary buds appear on the stock, the fructification condition exists in the later period, and the special seed coat color is convenient to distinguish.
3. The method utilizes the dark condition to culture the seedlings, can ensure that the hypocotyls of the seedlings grow quickly, starts grafting when the length of the hypocotyls is 6-8cm, can avoid the problems that the hypocotyls of stock materials cultured by common illumination are short (1-2cm) and are not easy to operate during grafting, and directly improves the grafting efficiency and the grafting survival rate. Compared with the method that only more than ten plants can be grafted in 1h when the hypocotyl is short, the method can graft more than 30 plants in 1h when the hypocotyl is long, and the working efficiency is improved by more than one time.
4. The method directly selects hypocotyls for grafting, removes cotyledons and the parts above, saves the trouble of removing lateral buds on the rootstock after transplanting in the field, and can better supply nutrition to the scion.
5. The stock material of the invention is in the nutrition pot, the survival grafted seedling transplanted to the field hardly damages the root, the seedling return is faster after transplanting, the survival rate can reach 100 percent, and the survival rate of the tissue culture seedling transplanted is about 80 percent. The grafted seedlings have vigorous roots, stronger field growth vigor than tissue culture seedlings, more bearing capacity than the tissue culture seedlings and better bearing plumpness.
6. The invention omits the complicated sterile grafting seedling culture and transplantation process, is not convenient to operate because the sterile grafting stock material needs to be disinfected and inoculated with the culture medium and is limited by the sterile condition during grafting, and needs to be exercised twice for transplantation after the survival of the sterile grafting seedling. The invention simplifies the experimental steps, is simple and convenient to operate and has better effect than aseptic grafting.
7. The method is simple, stable, reliable, easy to master and wide in application range. The method is suitable for grafting of peanut wild species materials, amphidiploid, transgenic plants and other tissue culture seedlings, and has certain reference value for other crops.
Drawings
FIG. 1 shows the observation results of 7d seedling-age of Yuhua 0215 cultured in dark in example 1.
FIG. 2 shows the observation of hypocotyl of peanut after cutting off cotyledons and parts thereof in dark culture of Yuhua 0215 of example 1 at 7d seedling age.
FIG. 3 shows the observation result of example 1 after grafting Yuhua 0215 with Yuhua 91027 d.
FIG. 4 is the observation result of example 1 after Yuhua 0215 is grafted with Yuhua 910214 d.
FIG. 5 is a field observation of examples 1 Yuhua 0215 and Yuza 9307 grafted seedlings.
FIG. 6 shows the setting of Yuhua 0215 and Yuza 9307 grafted seedlings and tissue culture seedlings in example 1.
Fig. 7 is the observation of example 2 after grafting of Yuhua 0215 to wild species a. cor 7 d.
FIG. 8 is a field observation of the grafted wild species Adard of Yuhua 0215 of example 2.
FIG. 9 shows the observation result of light-cultured 7d seedling age of Yuhua 0215 in comparative example 1.
FIG. 10 shows the setting of the Yuhua 0215 and Yuza 9307 grafted seedlings in comparative example 1.
FIG. 11 is an observation result of comparison example 2 after Yuhua 0215 is grafted with Yuhua 93077 d.
FIG. 12 shows the setting of Yuhua 0215 and Yuza 9307 grafted seedlings in comparative example 2.
Detailed Description
The following examples further illustrate the embodiments of the present invention in detail. The experimental procedures used in the following examples are, unless otherwise specified, conventional in the art or according to the conditions and procedures recommended by the manufacturer.
Example 1 grafting method of peanut cultivars tissue culture seedlings
A grafting method of peanut tissue culture seedlings comprises the following steps:
(1) preparation of stock material: selecting mature and plump seeds of black peanut Hehua 0215, soaking overnight for accelerating germination, planting the seeds in a nutrition pot (autoclaved vermiculite), watering thoroughly, placing in dark at 27 ℃ for culture, and selecting seedlings with hypocotyl length of 6-8cm as stock materials when two true leaves of the peanut seedlings are unfolded (about 7d) (see figure 1).
(2) Preparing the scions of the sterile tissue culture seedlings: and respectively taking Yuhua No. 22, Yuhua No. 9307 and Yuhua No. 9102 transgenic resistant seedlings as scions to carry out grafting (the height of a tissue culture seedling is more than 2 cm).
(3) Grafting: cutting off cotyledon and above parts of peanut seedling (stock), longitudinally cutting the middle of hypocotyl into a notch with depth of 0.8-1cm (see figure 2); cutting the lower end of the scion into a V-shaped wound corresponding to the cut, wherein the length of two inclined planes of the V-shaped wound is 0.8-1cm, and the cut is kept flat; inserting the lower end of the scion into the cut of the stock, aligning the lower end of the scion as much as possible, and tightly winding the scion with a Parafilm sealing film (forming a grafted seedling), so that the stock and the scion are tightly combined, and the wound healing is facilitated;
(4) culturing grafted seedlings: watering, placing the grafted seedling in a room (temperature is 27 ℃, humidity is 50%), preventing direct sunlight and covering a plastic film; and (3) removing the film after new leaves grow out (about 7 days), gradually culturing by light, namely, after new leaves grow out from the grafted seedlings, placing the grafted seedlings in the sun for 1 hour in the first day, increasing the illumination time day by day, increasing the illumination time by 1 hour every day, and transplanting the grafted seedlings to a field after 7 days, wherein the grafting survival rate reaches 100%. The specific culture conditions are shown in FIGS. 3 and 4, and FIGS. 3 and 4 show the culture conditions of Yuhua 0215 and Yuhua 9102 grafted seedlings;
(5) transplanting fields and managing: transplanting the grafted seedlings with good growth vigor to a field in the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; and after the transplanted seedlings completely survive, removing the sunshade net, performing normal field management, and transplanting the surviving grafted plants into the field to survive 100%. The specific growth conditions are shown in figure 5, and figure 5 shows the field growth conditions of Yuhua 0215 and Yuzao 9307 grafted seedlings.
The grafting survival rate and the transplanting survival rate of the tissue culture seedling of the cultivar of the embodiment are counted, and the results are shown in the following table.
The results in the table show that the grafting survival rate and the transplanting survival rate of the peanut tissue culture seedling grafting method can be improved.
The fruiting condition of the grafted seedling of this example was compared with that of the tissue cultured seedling, and the result is shown in fig. 6. The results in FIG. 6 show that the grafted seedling of the invention has vigorous root system, stronger field growth vigor than tissue culture seedling, more bearing capacity than tissue culture seedling and better fructification and plumpness.
Example 2 grafting method of peanut wild species tissue culture seedling
A grafting method of peanut tissue culture seedlings comprises the following steps:
(1) preparation of stock material: selecting mature and plump seeds of black peanut Hehua 0215, soaking overnight for accelerating germination, planting the seeds in a nutrition pot (autoclaved vermiculite), watering thoroughly, placing in dark at 27 ℃ for culture, and selecting seedlings with hypocotyl length of 6-8cm as stock materials when two true leaves of the peanut seedlings are unfolded (about 7 d).
(2) Preparing the scions of the sterile tissue culture seedlings: wild species (A.cor, A.dard and 30119) and progeny material (amphidiploid Am-L) are respectively used as scions to carry out grafting (the height of tissue culture seedlings is more than 2 cm).
(3) Grafting: cutting off cotyledons and parts above the cotyledons of peanut seedlings (rootstocks), and longitudinally cutting the middle of a hypocotyl into a notch with the depth of 0.8-1 cm; cutting the lower end of the scion into a V-shaped wound corresponding to the cut, wherein the length of two inclined planes of the V-shaped wound is 0.8-1cm, and the cut is kept flat; inserting the lower end of the scion into the cut of the stock, aligning the lower end of the scion as much as possible, and tightly winding the scion with a Parafilm sealing film (forming a grafted seedling), so that the stock and the scion are tightly combined, and the wound healing is facilitated;
(4) culturing grafted seedlings: watering, placing the grafted seedling in a room (temperature is 27 ℃, humidity is 50%), preventing direct sunlight and covering a plastic film; after new leaves (about 7d) grow, removing the film, and gradually culturing with light, namely, after the grafted seedlings grow new leaves, placing the grafted seedlings in the sun for 1h in the first day, increasing the illumination time day by day, increasing the illumination time by 1h every day, and transplanting the grafted seedlings to a field after 7d, wherein the specific culture conditions are shown in fig. 7, and fig. 7 shows the culture conditions of Yuhua 0215 and wild species A.Cor grafted seedlings;
(5) transplanting fields and managing: transplanting the grafted seedlings with good growth vigor to a field in the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; and after the transplanted seedlings completely survive, removing the sunshade net, and performing normal field management, wherein the specific growth conditions are shown in figure 8, and figure 8 shows the field growth conditions of the Yuhua 0215 and the wild species A.dard grafted seedlings.
The grafting survival rate and the transplanting survival rate of the wild species and the amphidiploid tissue culture seedlings of the embodiment are counted, and the results are shown in the following table.
The result shows that the grafting survival rate and the transplanting survival rate of the peanut tissue culture seedling grafting method can be improved, and the field of the transplanted seedling is strong.
Comparative example 1 grafting method of peanut cultivar tissue culture seedlings
A grafting method of peanut tissue culture seedlings comprises the following steps:
(1) preparation of stock material: selecting mature and plump seeds of black peanut Hehua 0215, soaking overnight for accelerating germination, planting the seeds in a nutrition pot (autoclaved vermiculite), watering thoroughly, placing in illumination culture at 27 ℃, and selecting hypocotyl seedlings as stock materials when two true leaves of the peanut seedlings are unfolded (about 7d) (see figure 9).
(2) Preparing the scions of the sterile tissue culture seedlings: and respectively taking Yuhua No. 22, Yuhua No. 9307 and Yuhua No. 9102 transgenic resistant seedlings as scions to carry out grafting (the height of a tissue culture seedling is more than 2 cm).
(3) Grafting: cutting off cotyledons and parts above the cotyledons of peanut seedlings (rootstocks), and longitudinally cutting the middle of a hypocotyl into a notch with the depth of 0.5-1 cm; cutting the lower end of the scion into a V-shaped wound corresponding to the cut, wherein the length of two inclined planes of the V-shaped wound is 0.5-1cm, and the cut is kept flat; inserting the lower end of the scion into the cut of the stock, aligning the lower end of the scion as much as possible, and tightly winding the scion with a Parafilm sealing film (forming a grafted seedling), so that the stock and the scion are tightly combined, and the wound healing is facilitated;
(4) culturing grafted seedlings: watering, placing the grafted seedling in a room (temperature is 27 ℃, humidity is 50%), preventing direct sunlight and covering a plastic film; and (3) removing the film after new leaves grow out (about 7 days), gradually culturing by light, namely, placing the grafted seedling in the sun for 1h in the first day after the new leaves grow out, increasing the illumination time day by day, and transplanting the grafted seedling into a field after the illumination time is increased by 1h every day and 7 days.
(5) Transplanting fields and managing: transplanting the grafted seedlings with good growth vigor to a field in the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; after the transplanted seedlings completely survive, the sunshade net is removed, normal field management is carried out, and the seedling bearing performance is good (figure 10).
The grafting survival rate and the transplanting survival rate of the tissue culture seedling of the cultivar of the embodiment are counted, and the results are shown in the following table.
The result shows that the grafting survival rate of the peanut tissue culture seedling grafting method in the comparison example is 87.5-95%, which is obviously lower than that of the grafting method in the invention (example 1), and the hypocotyls are shorter and almost all in soil, so that the grafting operation is inconvenient and the grafting efficiency is lower.
Comparative example 2 grafting method of peanut cultivar tissue culture seedlings
A grafting method of peanut tissue culture seedlings comprises the following steps:
(1) preparation of stock material: selecting mature and plump seeds of black peanut Hehua 0215, soaking overnight for accelerating germination, planting the seeds in a nutrition pot (autoclaved vermiculite), watering thoroughly, placing in illumination culture at 27 ℃, and selecting epicotyl seedlings as stock materials when two true leaves of the peanut seedlings are unfolded (about 7 d).
(2) Preparing the scions of the sterile tissue culture seedlings: and respectively taking Yuhua No. 22, Yuhua No. 9307 and Yuhua No. 9102 transgenic resistant seedlings as scions to carry out grafting (the height of a tissue culture seedling is more than 2 cm).
(3) Grafting: cutting off peanut seedlings (rootstocks) to cut off terminal buds and side buds, taking parts of epicotyl parts above cotyledonary nodes, and longitudinally cutting the middle of the epicotyl into a notch with the depth of 0.5-1 cm; cutting the lower end of the scion into a V-shaped wound corresponding to the cut, wherein the length of two inclined planes of the V-shaped wound is 0.5-1cm, and the cut is kept flat; inserting the lower end of the scion into the cut of the stock, aligning the lower end of the scion as much as possible, and tightly winding the scion with a Parafilm sealing film (forming a grafted seedling), so that the stock and the scion are tightly combined, and the wound healing is facilitated;
(4) culturing grafted seedlings: watering, placing the grafted seedling in a room (temperature is 27 ℃, humidity is 50%), preventing direct sunlight and covering a plastic film; and after new leaves (about 7 days) grow out, removing the film, gradually culturing by light, namely, after the grafted seedlings grow out new leaves, placing the grafted seedlings under the sunlight for 1h in the first day, increasing the illumination time day by day, increasing the illumination time by 1h every day, and transplanting the grafted seedlings into a field after 7 days, wherein the specific culture condition of the grafted seedlings is shown as figure 11, and figure 11 is an observation result after Yuhua 0215 is grafted with the Yuhua 93077 d.
(5) Transplanting fields and managing: transplanting the grafted seedlings with good growth vigor to a field in the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; and after the transplanted seedlings completely survive, removing the sunshade net, and performing normal field management. The fructification situation is shown in figure 12, and it can be seen that the fructification has fruits on the scion and also has fruits on the lateral bud of the rootstock.
The grafting survival rate and the transplanting survival rate of the tissue culture seedling of the cultivar of the embodiment are counted, and the results are shown in the following table.
The result shows that the transplanting survival rate of the peanut tissue culture seedling grafting method is 88.6-93.3%, which is obviously lower than that of the grafting method of the invention (embodiment 1), and because the lateral bud grows on the rootstock after the epicotyl grafted seedling is transplanted in the field, the lateral bud needs to be frequently removed, but the removal of the lateral bud is not thorough, so that the lateral bud of the rootstock grows vigorously, and the growth and the fructification conditions of the scion are influenced.
The foregoing description is only a preferred embodiment of the present invention, and various modifications and changes will occur to those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (8)
1. A grafting method of peanut tissue culture seedlings is characterized by comprising the following specific steps:
(1) culturing black peanut seeds under a dark culture condition, selecting seedlings with hypocotyl length of 6-8cm as stocks when two true leaves of peanut seedlings are unfolded, cutting off cotyledons and parts above the cotyledons of the peanut seedlings, longitudinally cutting the middle of the hypocotyls into cuts with the depth of 0.8-1 cm;
(2) taking the sterile tissue culture seedlings as scions, cutting the lower ends of the scions into V-shaped wounds corresponding to the cuts, and keeping the cuts flat; inserting the lower end of the scion into the cut of the stock, and tightly winding the scion with a Parafilm sealing film to enable the stock and the scion to be tightly combined to form a grafted seedling;
(3) watering, namely placing the grafted seedlings indoors to avoid direct sunlight and covering the grafted seedlings with a plastic film; removing the film after new leaves grow out, and gradually culturing with light;
(4) transplanting the grafted seedlings with good growth vigor to a field in a cloudy day or at the evening, watering thoroughly after the transplanting is finished, and covering a sunshade net for shading; and after the transplanted seedlings completely survive, removing the sunshade net, and performing normal field management.
2. The grafting method of peanut tissue culture seedlings according to claim 1, wherein the black peanut variety of step (1) is Yuhua 0215.
3. The grafting method of peanut tissue culture seedlings according to claim 1, wherein the black peanut seeds in the step (1) are cultured under dark conditions by the following steps: soaking black peanut seeds overnight for accelerating germination, then planting the seeds in a nutrition pot containing sterilized vermiculite, placing the seeds in the dark at 27 ℃ for culturing until two true leaves are unfolded, and selecting seedlings with the hypocotyl length of 6-8cm as stocks.
4. The method for grafting the peanut tissue culture seedling as claimed in claim 1, wherein the tissue culture seedling in the step (2) is a peanut wild species, an amphidiploid or a transgenic plant.
5. The method for grafting the peanut tissue culture seedling as claimed in claim 1, wherein the height of the tissue culture seedling in the step (2) is more than 2 cm.
6. The grafting method of peanut tissue culture seedlings according to claim 1, wherein the length of the two inclined planes of the V-shaped wound in the step (2) is 0.8-1 cm.
7. The grafting method of peanut tissue culture seedlings according to claim 1, wherein the indoor environment of the step (3) is 27 ℃ and 50% of humidity.
8. The grafting method of peanut tissue culture seedlings according to claim 1, wherein the grafted seedlings in the step (3) are placed under the sun for 1h in the first day after new leaves grow, the illumination time is increased by 0.5-1.5h day by day, and the seedlings can be transplanted to the field after 7 d.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102792892A (en) * | 2012-08-21 | 2012-11-28 | 青岛农业大学 | Method for directional screening of resistance body by peanut in-vitro mutation |
| KR20130019330A (en) * | 2011-08-16 | 2013-02-26 | (주)복슨로하스 | A method for production of peanut sprouts and the apparatus for that |
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| CN102388801B (en) * | 2011-07-27 | 2013-07-24 | 青岛农业大学 | Sterile grafting method of peanut tissue culture seedlings |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130019330A (en) * | 2011-08-16 | 2013-02-26 | (주)복슨로하스 | A method for production of peanut sprouts and the apparatus for that |
| CN102792892A (en) * | 2012-08-21 | 2012-11-28 | 青岛农业大学 | Method for directional screening of resistance body by peanut in-vitro mutation |
Non-Patent Citations (2)
| Title |
|---|
| Regeneration of fertile Arachis paraguariensis plants from callus and suspension cultures;P.E. STILL等;《Plant Cell，Tissue and Organ Culture》;19871231(第9期);37-43 * |
| 利用嫁接提高花生离体再生或转基因苗成活率的研究;李长生等;《中国农学通报》;20091020;第25卷(第20期);63-67 * |
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