CN107509535B - Efficient cleft grafting method for dicotyledon seedling and young branch - Google Patents

Efficient cleft grafting method for dicotyledon seedling and young branch Download PDF

Info

Publication number
CN107509535B
CN107509535B CN201710780422.9A CN201710780422A CN107509535B CN 107509535 B CN107509535 B CN 107509535B CN 201710780422 A CN201710780422 A CN 201710780422A CN 107509535 B CN107509535 B CN 107509535B
Authority
CN
China
Prior art keywords
scion
stock
cut
grafting
stem
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Active
Application number
CN201710780422.9A
Other languages
Chinese (zh)
Other versions
CN107509535A (en
Inventor
吴慎杰
秦丽霞
张换样
李静
李盛
焦改丽
竹梦婕
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Cotton Research Institute of Shanxi Academy of Agricultural Sciences
Original Assignee
Cotton Research Institute of Shanxi Academy of Agricultural Sciences
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Cotton Research Institute of Shanxi Academy of Agricultural Sciences filed Critical Cotton Research Institute of Shanxi Academy of Agricultural Sciences
Priority to CN201710780422.9A priority Critical patent/CN107509535B/en
Publication of CN107509535A publication Critical patent/CN107509535A/en
Application granted granted Critical
Publication of CN107509535B publication Critical patent/CN107509535B/en
Active legal-status Critical Current
Anticipated expiration legal-status Critical

Links

Images

Classifications

    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01HNEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H4/00Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
    • A01H4/008Methods for regeneration to complete plants
    • AHUMAN NECESSITIES
    • A01AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01GHORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G2/00Vegetative propagation
    • A01G2/30Grafting

Abstract

The invention discloses a high-efficiency cleft grafting method for dicotyledon seedlings and young branches. The method comprises the steps of adopting a stock with 1-2 true leaves (only 2 cotyledons are reserved) or a stock with 4-6 true leaves (2 cotyledons and 1-2 true leaves are reserved), and longitudinally splitting a stem and a lower edge of a petiole from the middle along a stem central axis from the lower edge of the cotyledon/true leaf petiole; cutting the scion into a V shape; the stem of the scion is inserted into the cut of the stock vertically. The cut of the rootstock simultaneously passes through the lower segment of the petiole and the middle axis of the stem, so that the scion is convenient to fix, the scion is naturally and closely combined with the rootstock, nutrient substances for photosynthesis of leaves can be delivered into the scion nearby through the petiole, and the rapid healing of the cut is promoted. The method combines the moisture preservation and the heat preservation in the seedling revival stage, the grafting survival rate can be stabilized to be more than 98 percent, the grafting survival rate can reach 100 percent in the suitable stock and scion matching stage, and the method is simple and convenient to operate, high in grafting speed and high in survival rate.

Description

Efficient cleft grafting method for dicotyledon seedling and young branch
Technical Field
The invention relates to a cleft grafting method of plant seedlings and young branches, in particular to a high-efficiency cleft grafting method of dicotyledonous plant seedlings and young branches, belonging to the technical field of agriculture.
Background
Genetic engineering and cell engineering are important means for genetic improvement of cotton at present, cotton tissue culture regenerated plants are the basis of genetic improvement application of a transgenic technology in cotton, however, most root systems of test-tube seedlings of the transgenic regenerated cotton obtained through somatic embryogenesis are poor in development and are expressed as rootless seedlings, brown root seedlings, abnormal seedlings (such as clumpy seedlings) and the like, the regenerated plants with normal root systems are transplanted into soil or vermiculite, the survival rate is extremely low (usually only 10-20%), the regeneration plants grow slowly, and the application of the cotton genetic improvement technology is greatly limited.
The grafting is a vegetative hybrid cultivation technology, usually, sea island cotton with developed root system and strong disease resistance is selected as a stock, and a transgenic regenerated cotton plant is directly grafted to the stock growing vigorously, so that nutrient substances in the stock and a scion body are exchanged, the disease resistance of the scion is improved, the adaptability of the scion to the external environment is enhanced, the growth and development are accelerated, the scion is mature in advance, and the survival rate of the scion is greatly improved.
The establishment and the perfection of the grafting system are beneficial to the large-scale preservation and breeding of genetic segregation population, conventional cotton plants and rare germplasm resources, and are beneficial to accelerating the wide application of modern biotechnology in cotton genetic improvement and greatly accelerating the breeding process of transgenic cotton.
According to the relevant data, some researchers respectively adopt a combination grafting method, a cleft grafting method and a bark grafting method to carry out cotton grafting, and the survival rate of cotton plants is improved to a certain extent. But for regenerated plant grafts:
(1) the conventional cleft grafting method is characterized in that the head of a cotyledon stalk is picked and the core is removed, the cut is vertical to the cotyledon stalk, the cotyledon can be sloughed towards two sides under the action of gravity, and the scion is difficult to insert and fix. Some researchers also leave a section of epicotyl at the upper end of a cotyledon stalk and longitudinally split, however, the method is not convenient for supplying nutrition to the scion by the rootstock leaves;
(2) the rootstock required by the 'bark grafting method' is larger;
(3) the 'conjugal grafting' method has less cambium contact surface and less nutrient exchange between the stock and the scion body.
Therefore, the grafting technology of cotton still needs to be studied in a system, and a perfect cotton grafting technology system also needs to be established.
Disclosure of Invention
The invention aims to provide a high-efficiency cleft grafting method for dicotyledon seedlings and young branches, which is simple and convenient to operate and can effectively improve the grafting survival rate.
In order to achieve the above purpose, the invention adopts the following technical scheme:
the efficient cleft grafting method of dicotyledon seedling and young branch is characterized by comprising the following steps:
step 1: for the rootstocks with 1-2 true leaves, only 2 cotyledons are reserved, and stems and leaves at the positions above 2 cotyledons are cut off; for the rootstock with 4-6 true leaves, reserving 2 cotyledons and 1-2 true leaves of the rootstock, cutting off stems and leaves at the upper part, starting from a position 5mm below a cotyledon petiole/true leaf petiole, longitudinally splitting a stem and the lower edge of the petiole from the middle along a stem center axis, wherein the length of a cut is 1.5-2 cm;
step 2: keeping 2-4 young and tender true leaves and growing points at the top end of the scion, wherein the length is 3-5 cm, cutting 1 knife at each side of the lower section of the stem of the scion, sharpening, and cutting the surface into a V shape;
step 3: vertically inserting the stem of the scion into the cut of the stock, enabling the petioles of the stock to promote the scion to be tightly combined with the cut of the stock, and then winding and fixing the scion and the stock by using a rope;
step 4: tightly wrapping the whole pot of plants with a plastic film, and placing the pot of plants under a shed;
step 5: when the grafted plant passes through the seedling recovering period and the scion leaf returns to be flat and starts to grow, the opening of the plastic film wrapping the plant is gradually ventilated, ventilated and dehumidified, and finally the film is uncovered and the fixture is removed, thus completing grafting.
The efficient cleft grafting method of dicotyledon seedling and young branch is characterized in that in Step1, the variety of the rootstock is sea island cotton, black seed pumpkin, Indian pumpkin or bottle gourd.
The efficient cleft grafting method for dicotyledon seedling and young branch is characterized in that in Step1, the incision is 1.5cm long.
The efficient cleft grafting method for the dicotyledon seedlings and the young branches is characterized in that in Step3, the scion and the rootstock are wound and fixed by a plastic rope from the joint of the 2-leaf blade stalk of the rootstock and the scion to the stem section of the rootstock.
The efficient cleft grafting method for the dicotyledon seedling and the young branch is characterized in that in Step3, a cut part is clamped by a grafting clamp, and the scion and the stock are tightly combined together.
The efficient cleft grafting method for the dicotyledon seedlings and the young branches is characterized in that in Step4, the environmental humidity is controlled to be more than or equal to 80%, and the environmental temperature is controlled to be 25-28 ℃.
The invention has the advantages that:
(1) the cut of the stock simultaneously passes through the lower segment of the petiole and the middle shaft of the stem, the scion can be conveniently fixed by utilizing the fixing action of the petiole, the scion is naturally and closely combined with the stock, nutrient substances with photosynthesis of leaves can be delivered into the scion nearby through the petiole, the rapid healing of a connector is promoted, the whole grafting operation is simple, the grafting speed is high, the survival rate is high, and the survival rate can reach more than 98% when the plant is grafted by adopting the method disclosed by the invention through statistics;
(2) the cleft grafting method is not only suitable for grafting the cotton plant seedlings and the young branches regenerated by transgenic and tissue culture, but also suitable for grafting many other dicotyledonous plant seedlings and young branches, such as cucumber, muskmelon, watermelon, apple seedlings and the like, and has a wide application range.
Drawings
FIG. 1 is a schematic representation of the cut of a rootstock (1-2 true leaves grown);
FIG. 2 is a schematic view of the cutting face of a scion;
FIG. 3 is a schematic illustration of the insertion of a scion into a cut of a rootstock;
FIG. 4 is a schematic winding diagram of scion and rootstock;
FIG. 5 is a schematic representation of the cut of the rootstock (with 4-6 true leaves growing);
figure 6 is a schematic illustration of the insertion of the scion into the cut of the rootstock.
Detailed Description
The invention is described in detail below with reference to the figures and the embodiments.
Example 1 grafting of transgenic regenerated Cotton plants on Gossypium barbadense with 1-2 true leaves
In late autumn, winter and spring, the temperature of the greenhouse is proper, grafting is directly carried out in the greenhouse, and the optimal temperature of the greenhouse is controlled to be 28 +/-1 ℃; the temperature is higher in summer and early autumn, and grafting is carried out in 50% of sunshades.
Stock: the potted cotton is cultivated into 20 pots of 5 plants of island cotton seedlings, the total number of 100 plants, when cotyledons are completely unfolded, the seedlings are transplanted into plastic pots with the diameter of about 20cm, and one plant in each pot is used as a stock.
Scion grafting: tissue culture sterile transgenic cotton seedlings are used as scions.
Step 1: one day before grafting, the island cotton (cotton seedlings with 1-2 true leaves and 2 complete cotyledons) planted in the flowerpot is watered with enough water, only 2 cotyledons of the stock are reserved, a sharp blade is used for cutting off stems and leaves at the positions above 2 cotyledons by sticking to cotyledonary petioles, then the stems and the lower edges of the petioles are longitudinally split from the middle along the central axis of the stems from the position 5mm away from the lower edges of the cotyledonary petioles, the cut is smooth, and the cut length is 1.5cm, as shown in figure 1.
Step 2: the root of a cotton transgenic regeneration plant is removed (a scion is obtained), 2 tender true leaves and growing points at the top end of the scion are reserved, the length of the scion is controlled to be 3-5 cm, redundant stem leaves and roots at the lower part of the scion are removed, then 1 cutter is respectively cut on two sides of the lower section of a stem of the scion, the scion is sharpened, the cut surface is in a V shape, and the cut surface length is 1.5-2 cm, as shown in figure 2.
Step 3: slightly twisting the cut part of the stock by using a thumb and a forefinger to open the cut, vertically and downwards inserting the cut stem section of the scion into the cut of the stem section of the stock, wherein the cut penetrates through the leaf stalk and the stem section (different from the reported direct splitting of the stem section), as shown in figure 3, the leaf stalk has a fixing function, so that the scion and the cut of the stock can be naturally and tightly combined, the fixation of a rope and a grafting clip is facilitated, further the photosynthetic product of the leaves can be transported to the cut nearby, the wound healing and the scion growth are promoted, and the survival rate is remarkably improved; after the scions are inserted, the scions and the rootstocks are wound and fixed from bottom to top obliquely by using plastic ropes from the combination parts of the 2 leaves and the stalks of the rootstocks to the stem sections of the rootstocks, and then the scions are wound and knotted downwards, as shown in figure 4.
Step 4: taking 2 iron wires with proper length, crossing the top ends of the 2 iron wires, inserting the two ends into the edge of the flowerpot to form a small arch of about 25-30 cm, tightly wrapping the flowerpot and the iron wires (namely wrapping the whole pot plant) by using a plastic film, tightly binding the periphery of the flowerpot and the iron wires for moisturizing, placing the flowerpot and the iron wires under a pergola to prevent the graft from withering, controlling the environmental humidity to be more than or equal to 80% and controlling the environmental temperature to be 28 +/-1 ℃ at the same time when the grafted plant enters the seedling revival stage.
Step 5: when the grafted plant passes through the seedling recovering period, the scion blade recovers flattening and starts to grow, and the plastic film opening for wrapping the plant is gradually ventilated and dehumidified, which specifically comprises the following steps: and (3) on 4-7 days after grafting, drilling a round hole with the thickness of a finger on the top end of the film for ventilation so as to prevent the scion from mildewing, then, growing 1-2 new leaves on the scion, uncovering the film about 10-12 days after grafting, untying around 15 days after grafting, healing the junction of the stock and the scion, enabling the grafted cotton plant to survive completely, enabling the survival rate to reach 100%, and transplanting the grafted cotton plant to a field or a pot after one week.
The cut of the stock simultaneously passes through the lower segment of the petiole and the middle shaft of the stem, the scion can be conveniently fixed by utilizing the fixing effect of the petiole, the scion is naturally and closely combined with the stock, nutrient substances with photosynthesis of leaves can be delivered into the scion nearby through the petiole, the rapid healing of a connector is promoted, the whole grafting operation is simple, the grafting speed is high, and the survival rate is high.
Statistics shows that when the method is adopted to graft transgenic regenerated cotton plants on the sea island cotton, the survival rate reaches 99.8 percent.
Example 2 grafting of transgenic regenerated Cotton plants on Gossypium barbadense with 4-6 leaves
Stock: the potted cotton is cultivated into 20 pots of 5 plants of island cotton seedlings, the total number of 100 plants, when cotyledons are completely unfolded, the seedlings are transplanted into plastic pots with the diameter of about 20cm, and one plant in each pot is used as a stock.
Scion grafting: sterile transgenic cotton seedlings in culture medium of the culture room are used as scions.
Step 1: one day before grafting, the island cotton (cotton seedlings with 4-6 true leaves growing) planted in the flowerpot is watered with enough water, 2 cotyledons and 1-2 true leaves of the stock are reserved, the stem and the leaf at the upper part are cut off by a sharp blade, then the stem and the lower edge of the leaf stalk are longitudinally split from the middle along the stem center axis from the position 5mm away from the lower edge of the leaf stalk of the true leaf, the cut is smooth, and the cut length is 1.5cm, as shown in figure 5.
Step 2: the root of a cotton transgenic regeneration plant is removed, 2 tender true leaves and growing points at the top end of a scion are reserved, the length of the scion is controlled to be 3-5 cm, redundant stems and leaves and roots at the lower part of the scion are removed, then 1 knife is respectively cut at two sides of the lower section of a stem of the scion, the scion is sharpened, the cut surface is V-shaped, and the cut surface length is 1.5-2 cm, as shown in figure 2.
Step 3: slightly twisting the cut part of the stock by using a thumb and a forefinger to open the cut, vertically inserting the stem section of the cut scion into the cut of the stem section of the stock downwards, and enabling the cut to penetrate through the petioles and the stem section, as shown in figure 6; after the scions are inserted, the scions and the rootstocks are wound and fixed from bottom to top obliquely by plastic ropes from the joints of the true leafstalks of the rootstocks and the scions to the stem sections of the rootstocks, and then the scions are wound and knotted downwards.
Step 4: taking 2 iron wires with proper length, crossing the top ends of the 2 iron wires, inserting the two ends into the edge of the flowerpot to form a small arch of about 25-30 cm, tightly wrapping the flowerpot and the iron wires (namely wrapping the whole pot plant) by using a plastic film, tightly binding the periphery of the flowerpot and the iron wires for moisturizing, placing the flowerpot and the iron wires under a pergola to prevent the graft from withering, controlling the environmental humidity to be more than or equal to 80% and controlling the environmental temperature to be 28 +/-1 ℃ at the same time when the grafted plant enters the seedling revival stage.
Step 5: when the grafted plant passes through the seedling recovering period, the scion blade recovers flattening and starts to grow, and the plastic film opening for wrapping the plant is gradually ventilated and dehumidified, which specifically comprises the following steps: and (3) on 4-7 days after grafting, drilling a round hole with the thickness of a finger on the top end of the film for ventilation so as to prevent the scion from mildewing, then, growing 1-2 new leaves on the scion, uncovering the film about 10-12 days after grafting, untying around 15 days after grafting, healing the junction of the stock and the scion, enabling the grafted cotton plant to survive completely, enabling the survival rate to reach 100%, and transplanting the grafted cotton plant to a field or a pot after one week.
Statistics shows that when the method is adopted to graft transgenic regenerated cotton plants on the sea island cotton, the survival rate reaches 99.5 percent.
Cotton seedlings of island cotton with 1-6 main leaves can be used as rootstocks, and when the scions are tissue culture seedlings with 4-5 main leaves and stem sections with uniform thickness, the optimal seedling age period of matching the rootstocks and the scions is obtained. Through statistics, the survival rate of grafting can reach 100% by adopting the combination mode.
Example 3 grafting of cucumber seedlings on Black-seed pumpkin having 1-2 true leaves
In early spring, 100 pots of black-seed pumpkin and cucumber are respectively sown in the nutrition pots, 2-3 plants are planted in each pot, and 1 plant growing robustly is reserved in each pot when cotyledons are completely unfolded.
Stock: 1-2 true leaves grow out from the pumpkin seedling, and the 1 st true leaf can be used as the stock when the length is about 1 cm.
Scion grafting: after sowing for 10 days, the seed leaves of the cucumber seedlings are flattened, and 1 true leaf and a growing point grow out, so that the cucumber seedlings can be used as scions.
Step 1: one day before grafting, the pumpkin seedlings (seedlings with 1-2 true leaves and 2 complete cotyledons) planted in the nutrition pot are watered with enough water, only 2 cotyledons of the stock are reserved, sharp blades are used for sticking cotyledonary petioles to cut off stems and leaves at the positions above 2 cotyledons, then the stems and the lower edges of the petioles are longitudinally split from the middle along the central axis of the stems from the position 5mm away from the lower edges of the cotyledonary petioles, the cuts are smooth, and the length of the cuts is about 1.5 cm.
Step 2: removing the roots of the cucumber seedlings with the cotyledons flattened (to obtain the scions), keeping 1 tender true leaf and a growing point at the top end of the scions, controlling the length of the scions to be 3-5 cm, removing redundant stem leaves and roots at the lower parts, then respectively cutting 1 knife at two sides of the lower section of a stem of the scions, sharpening, wherein the cut surfaces are V-shaped, and the cut surface length is about 1.5 cm.
Step 3: slightly twisting the cut part of the stock by using a thumb and a forefinger to open the cut, vertically and downwards inserting the stem section of the cut scion into the cut of the stem section of the stock, and enabling the cut to penetrate through the petioles and the stem section; after the scions are inserted, the scions and the rootstocks are wound and fixed from bottom to top obliquely by plastic ropes from the joints of 2 cotyledon petioles of the rootstocks and the scions to stem sections of the rootstocks, and then the scions are wound and knotted downwards.
Step 4: spraying water, and placing in a small moisture-keeping arched shed. And maintaining 95% humidity within 3 days after grafting, wherein the temperature is 25-28 ℃ in the daytime and 18-20 ℃ at night. Ventilating slightly after 4 days, and removing the film to harden the seedlings after 8 days. The survival rate reaches 100 percent, and the three leaves can be planted in the one-heart period after about 25 days.
According to statistics, when cucumber seedlings are grafted on the black-seed pumpkin seedlings by the method, the survival rate reaches 99.6%.
Example 4 grafting of melon seedlings on pumpkin from India
Respectively sowing 100 pots of Indian pumpkin (winter squash) and melon (New Yumei) in the nutrition pot, wherein 2-3 plants are planted in each pot, and placing the pots in a greenhouse. And when the cotyledon is completely unfolded, 1 plant with stronger growth is reserved in each pot. The pumpkin of Indian province is planted 1 week earlier than the melon.
Stock: the pumpkin seedlings in India grow 1-2 main leaves, 2 sub-leaves are fully flattened, the 1 st main leaf is expanded (namely, broken core), the height of the seedlings is 6-8 cm, and the seedlings can be used as stocks when stems are not empty.
Scion grafting: after the first true leaf of the melon leaf is flattened and the head is exposed, the melon leaf can be used as a scion.
Step 1: taking the stock seedling to remove true leaves and growing points, reserving 2 intact cotyledons, longitudinally splitting the lower edges of the stems and the petioles from the middle along the middle axis of the stem from the position 5mm below the lower edges of the petioles of the cotyledons by using a sharp knife, and ensuring that the cut is smooth and the length of the cut is about 1.5 cm.
Step 2: and (3) taking the scion seedlings, and downwards cutting the scion seedlings into double-sided wedges at the positions of l-1.5 cm below cotyledons, wherein the length of each wedge is 1-1.5 cm, and the wedge surface is perpendicular to the extension lines of the two cotyledons, so that the rootstock cotyledons can support the scion seedlings.
Step 3: the wedge surface of the scion is inserted into the cut of the stock, the scion is parallel to the surface of the stock, and then the grafting clip prepared in advance is used for clamping the interface to enable the joint surface to be tightly combined.
Step 4: placing in a small moisture-keeping arched shed, shading with a sunshade net, gradually increasing illumination after 3 days, ventilating, and removing the binding fixture after 7 days. And (3) about 7 days before field planting, reducing the temperature to harden the seedlings so as to adapt to the environmental conditions after field planting.
Through statistics, when the method is adopted to graft the melon (New Yumeiren) seedlings on the Indian pumpkin seedlings, the survival rate reaches 99.7 percent.
Example 5 grafting of Young watermelon shoot on bottle gourd
And (3) respectively sowing 100 pots of potted bottle gourds and watermelons (xiabao) in the nutrition pot, wherein 2-3 plants are planted in each pot, and placing the pots in a greenhouse. And when the cotyledon is completely unfolded, 1 plant with stronger growth is reserved in each pot. The bottle gourd is sowed 10 days earlier than the watermelon, when true leaves of the bottle gourd appear, the terminal bud is removed, and the optimal growth temperature is 25 ℃.
Stock: the bottle gourd seedlings grow into 2-3 main leaves, and the 2 sub-leaves are fully flattened to be used as stocks.
Scion grafting: the watermelon cotyledon is flattened, and the first true leaf is exposed, namely, the core is broken, and then the first true leaf can be used as a scion.
Step 1: taking the stock seedling to remove true leaves and growing points, reserving 2 intact cotyledons, longitudinally splitting the lower edges of the stems and the petioles from the middle along the middle axis of the stem from the position 5mm below the lower edges of the petioles of the cotyledons by using a sharp knife, wherein the cut is smooth and is about 1.5cm long, and the blade is not pulled out temporarily.
Step 2: and (3) taking the scion seedlings, and downwards cutting the scion seedlings into double-sided wedges at the positions of l-1.5 cm below cotyledons, wherein the length of each wedge is 1-1.5 cm, and the wedge surface is perpendicular to the extension lines of the two cotyledons, so that the rootstock cotyledons can support the scion seedlings.
Step 3: and removing the blade, immediately inserting the wedge surface of the cut scion into the cut of the stock to enable the scion to be flush with the surface of the stock, and clamping the interface by using a prepared grafting clamp to enable the joint to be tightly combined.
Step 4: placing the greenhouse in a small moisture-preserving arched shed, shading by a shading net, sealing the shed room, keeping the indoor relative humidity above 90%, and keeping the room temperature at 24-26 ℃. And gradually increasing the time of light exposure for 3-5 days, and removing the shading hardening seedlings after 10 days. Ventilation is reduced as much as possible 2-3 days after grafting, and ventilation is only required 1-2 times per day (in the morning or evening).
According to statistics, when the method is adopted to graft watermelon (xiabao) seedlings on bottle gourd seedlings, the survival rate reaches 99.5%.
Therefore, the cleft grafting method is not only suitable for grafting the transgenic (and tissue culture) regenerated cotton plant seedlings (and young branches), but also suitable for grafting many other dicotyledonous plant seedlings and young branches, such as cucumber, melon, watermelon and the like, and has a wide application range.
It should be noted that the above-mentioned embodiments do not limit the present invention in any way, and all technical solutions obtained by using equivalent alternatives or equivalent variations fall within the protection scope of the present invention.

Claims (6)

1. The efficient cleft grafting method of dicotyledon seedling and young branch is characterized by comprising the following steps:
step 1: for the stock growing with 1-2 true leaves, only 2 cotyledons of the stock are reserved, stems and leaves at the positions above the 2 cotyledons are cut off, for the stock growing with 4-6 true leaves, 2 cotyledons and 1-2 true leaves of the stock are reserved, stems and leaves at the upper part are cut off, then, the stem and the lower edge of the petiole of the cotyledons/the lower edge of the petiole of the true leaves are longitudinally split from the middle along the stem center axis, and the length of the cut is 1.5-2 cm;
step 2: keeping 2-4 young and tender true leaves and growing points at the top end of the scion, wherein the length of the true leaves and growing points is 3-5 cm, cutting 1 knife at each side of the lower section of the stem of the scion, sharpening, and making the cut surface in a V shape;
step 3: vertically inserting the stem of the scion into the cut of the stock, enabling the petioles of the stock to promote the scion to be tightly combined with the cut of the stock, and then winding and fixing the scion and the stock by using a rope;
step 4: tightly wrapping the whole pot of plants by using a plastic film and placing the pot of plants under a shed;
step 5: when the grafted plant passes through the seedling recovering period and the scion leaf returns to be flat and starts to grow, the opening of the plastic film wrapping the plant is gradually ventilated, ventilated and dehumidified, and finally the film is uncovered and the fixture is removed, thus completing grafting.
2. The method for efficient cleft grafting of dicotyledonous plant seedlings and young shoots according to claim 1, wherein in Step1, the variety of the rootstock is Gossypium barbadense, Cucurbita schatana, Cucurbita moschata, or Cucurbita pepo.
3. The method for efficient cleft grafting of dicot seedlings and young shoots according to claim 1, wherein said incision is 1.5cm in Step 1.
4. The method for efficient cleft grafting of dicotyledonous plant seedlings and young branches according to claim 1, wherein in Step3, the scion and the rootstock are wound and fixed by a plastic rope from the junction of the 2-leaf stalk of the rootstock and the scion to the stem section of the rootstock.
5. The method for efficient cleft grafting of dicotyledonous plant seedlings and young branches according to claim 1, wherein at Step3, the cut part is clamped by a grafting clip, and the scion and the rootstock are tightly combined.
6. The efficient cleft grafting method for dicotyledonous plant seedlings and young branches according to claim 1, characterized in that in Step4, the environmental humidity is controlled to be more than or equal to 80%, and the environmental temperature is controlled to be 25-28 ℃.
CN201710780422.9A 2017-09-01 2017-09-01 Efficient cleft grafting method for dicotyledon seedling and young branch Active CN107509535B (en)

Priority Applications (1)

Application Number Priority Date Filing Date Title
CN201710780422.9A CN107509535B (en) 2017-09-01 2017-09-01 Efficient cleft grafting method for dicotyledon seedling and young branch

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
CN201710780422.9A CN107509535B (en) 2017-09-01 2017-09-01 Efficient cleft grafting method for dicotyledon seedling and young branch

Publications (2)

Publication Number Publication Date
CN107509535A CN107509535A (en) 2017-12-26
CN107509535B true CN107509535B (en) 2020-09-01

Family

ID=60725041

Family Applications (1)

Application Number Title Priority Date Filing Date
CN201710780422.9A Active CN107509535B (en) 2017-09-01 2017-09-01 Efficient cleft grafting method for dicotyledon seedling and young branch

Country Status (1)

Country Link
CN (1) CN107509535B (en)

Families Citing this family (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108575364A (en) * 2018-04-02 2018-09-28 山东伟丽种苗有限公司 Two points one line graftings
CN110612882B (en) * 2019-11-08 2022-02-01 河南大学 Grafting breeding method for Bikstan cotton
CN112715185B (en) * 2021-02-02 2022-11-04 成都大学 Buckwheat grafting method
CN114568139B (en) * 2022-03-08 2023-03-21 中国热带农业科学院热带作物品种资源研究所 Grafting method for dragon fruit seedling sowing seedlings
CN114766224B (en) * 2022-06-07 2023-09-15 青岛金妈妈农业科技有限公司 Method for rapidly propagating cucumber stock material seeds

Family Cites Families (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1229140A (en) * 1999-03-30 1999-09-22 中国科学院遗传研究所 Producing method for transfer-gen plant
CN101558736A (en) * 2009-05-14 2009-10-21 浙江大学 Method for breeding cotton transgenic seedlings
CN101642022A (en) * 2009-08-25 2010-02-10 河南省农业科学院 Method for culturing cotton resisting micro-bundle disease
CN101878708B (en) * 2009-12-24 2012-05-23 江苏沿海地区农业科学研究所 Asexual grafting method of island cotton and upland cotton
CN101855969B (en) * 2010-06-03 2011-07-20 山东棉花研究中心 Grafting root-planting method for cotton
CN102893813B (en) * 2012-10-25 2013-10-23 山西省农业科学院棉花研究所 Grafting technique for excrescent buds of regenerated plants of genetically modified cotton
CN105265184A (en) * 2014-06-18 2016-01-27 闵建华 Cotton grafting splice grafting method

Also Published As

Publication number Publication date
CN107509535A (en) 2017-12-26

Similar Documents

Publication Publication Date Title
CN107509535B (en) Efficient cleft grafting method for dicotyledon seedling and young branch
CN1729746A (en) Cotton grafting and fit grafting method
CN113826513B (en) Asexual rapid propagation seedling raising method for celastrus orbiculatus
CN104737831A (en) Grape green branch stock hard branch grafting method
CN104041336A (en) High-branch quickly-cultivated seedling formed by grafting cercis canadensis onto cercis gigantea stock
CN107646576B (en) Watermelon grafting seedling method
CN107969238A (en) The breeding method of color silk flosssilk wadding wood
CN108934513B (en) Bud grafting method of apocarya in spring
CN106954467B (en) Summer grafting and seedling raising method for crape myrtle
CN114009232B (en) Tea tree grafting method and application thereof
CN115885710A (en) Rapid and efficient cultivation method for bird's nest fruit
CN113207524A (en) Pruning method and planting method for birch-leaf pear seedlings
CN106134806B (en) Grafting propagation method for natural hybrid populus grey in whole growing season
CN106613690B (en) Management method of walnut tree after grafting
CN105265184A (en) Cotton grafting splice grafting method
CN105359920A (en) Twig-grafting breeding method for buxus sinica
CN101558736A (en) Method for breeding cotton transgenic seedlings
CN111183807A (en) Method for breeding clone seedlings by cutting lateral buds of cucumbers
CN109526431A (en) Apocarya container seedling grafting of tender branch method
CN106688627A (en) Cotton grafting method
CN109819795A (en) A kind of acer monoes cuttage and seedling culture method
CN115474503B (en) Method for cultivating ornamental seedlings by double-binding combined cutting for long cutting surface
CN114175934B (en) Green seedling grafting method for schisandra chinensis
CN114766224B (en) Method for rapidly propagating cucumber stock material seeds
CN114009233B (en) Tea tree grafting seedling raising method and application thereof

Legal Events

Date Code Title Description
PB01 Publication
SE01 Entry into force of request for substantive examination
SE01 Entry into force of request for substantive examination
CB03 Change of inventor or designer information
CB03 Change of inventor or designer information

Inventor after: Wu Shenjie

Inventor after: Qin Lixia

Inventor after: Zhang Huanyang

Inventor after: Li Jing

Inventor after: Li Sheng

Inventor after: Jiao Gaili

Inventor after: Zhu Mengjie

Inventor before: Qin Lixia

Inventor before: Wu Shenjie

Inventor before: Zhang Huanyang

Inventor before: Li Jing

Inventor before: Li Sheng

Inventor before: Jiao Gaili

Inventor before: Zhu Mengjie

GR01 Patent grant
GR01 Patent grant