CN103782686B - A kind of watermelon plantlet in vitro grafting method - Google Patents
A kind of watermelon plantlet in vitro grafting method Download PDFInfo
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- CN103782686B CN103782686B CN201410065759.8A CN201410065759A CN103782686B CN 103782686 B CN103782686 B CN 103782686B CN 201410065759 A CN201410065759 A CN 201410065759A CN 103782686 B CN103782686 B CN 103782686B
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Abstract
The invention discloses a kind of watermelon plantlet in vitro grafting method.Step comprises: stock preparation, scion preparation, grafting, grafting cultivation, grafting are transplanted.Stock seed is seeded in nutrient discs after vernalization, two leaves wholeheartedly time can be used for grafting; The watermelon plantlet in vitro that 2cm ~ 3cm is high does scion after domestication; Tip cutting method carries out grafting; Grafting is incubated in time, moisturizing, shading are cultivated, and in time removed the sprout tillers of stock; Tool two leaf grafting wholeheartedly can transplant land for growing field crops, and field management is with common grafting seedling.Plantlet in vitro of the present invention, without the need to taking root, directly obtains Grafted Watermelon Seedlings and can overcome the take root problem of difficulty or instability of watermelon plantlet in vitro and save time again; Can carry out in the anniversary, manual control condition; Cycle is short, survival rate is high, easy to implement the method, easy and simple to handle, reduces Grafted Watermelon Seedlings production cost; Solve the regeneration plant existed at present in the training of watermelon group to take root undesirable and that transplanting survival rate is low problem.
Description
Technical field
The present invention relates to graft technology field, more specifically relate to a kind of watermelon plantlet in vitro grafting method.Cost is low, and graft survival rate is high.
Background technology
Watermelon [
citrullusLanatus(Thunb.) Matsum.etNakai] Curcurbitaceae, Citrullus, the annual herbaceous plant that overgrows.Originate in Africa, be one of large fruit in the world five, in Chinese national economy, occupy critical role.Grafting is the unique effective ways solving the soil-borne diseases such as watermelon blight at present, although watermelon seed grafting seedling can large-scale production and popularization in producing at present, the kind cost for aspermous watermelon and seed price costliness is higher.Meanwhile, although watermelon plantlet in vitro is in growth coefficient, reproduction speed and quantitatively have more advantage, seeling industry cost can be reduced, but due to tissue culture regeneration seedling rooting undesirable and after taking root transplanting survival rate not high yet, thus almost do not apply in actual production, this also becomes the large obstacle carrying out the correlative studys such as watermelon cell engineering, gene engineering.
Plantlet in vitro graft technology can effectively overcome the above problems, and such as, has researcher to make stock with diploid R. pseudoacacia seedling from seed, and tetraploid locust plantlet in vitro carries out grafting do scion and obtains regeneration plant, really realizes high efficiency quick breeding.The regeneration plant stabilization characteristics of genetics that the method is cultivated, plant is healthy, and grow vigorous, growing-seedling period is short, and seedling is fast, transplanting survival rate more than 90%.
On the basis of grasping watermelon plantlet in vitro production technology, the present invention proposes with watermelon plantlet in vitro as scion, take seedling as the graft technology of stock, solve the regeneration plant existed at present to take root and the problem of transplant survival difficulty, watermelon plantlet in vitro is used for grafting to produce, simultaneously for the researchs such as the research of Grafting watermelon correlation theory, breed improvement, cell engineering provide technical support.
Summary of the invention
The object of the present invention is to provide a kind of watermelon plantlet in vitro grafting method, with watermelon plantlet in vitro for scion, with the graft technology that cucurbit or pumpkin seedling are stock, solve the regeneration plant existed at present in the training of watermelon group take root and transplant the problem of difficulty, watermelon plantlet in vitro is produced, simultaneously for the researchs such as the research of Grafting watermelon correlation theory, breed improvement, cell engineering provide technology to ensure for grafting.
To achieve these goals, the present invention is by the following technical solutions:
A grafting method for watermelon plantlet in vitro, the steps include:
A, stock prepare, and stock seed is seeded in nutritive cube through vernalization, are used for grafting during the true leaf of the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon with five points of coin-size;
B, scion prepare, and growth medium grows to 25 days-30 days, grows to the plantlet in vitro of 2cm-3cm, and hardening of uncapping in 20 DEG C of-25 DEG C of greenhouse by solar heats is after 2 days-4 days, select that growing way is consistent, the plantlet in vitro of band 4 blades does scion;
C, the grafting of tip cutting method, first erase rootstock seedling the 1st true leaf base portion during grafting together with growing point, then with through 1%KnMO
4the tip of the bamboo let of solution disinfection is from growing point position to another cotyledon petiole base portion oblique cutting at 45 °, scion seedling is at distance plantlet in vitro foot blade 1cm place blade wedging, entered by the scion oblique cutting cut in the jack that bamboo let on stock puts on, stock is exposed at tip;
D, grafting are cultivated, grafting heat and moisture preserving, and seedling dish plastic film wraps up, and shading in first 3 days is cultivated, and throws off plastic film after 7 days, within 14 days, remove the sprout tillers of stock later;
E, grafting are transplanted, and grafting grafting tool two leaf of 22 days-28 days wholeheartedly can transplant land for growing field crops.
Described stock is cucurbit or pumpkin seedling.
Described stock prepares, and comprises the steps:
A. presoaking and germinating, with the water seed soaking of 55 DEG C, continuous stirring, until water temperature drops to 30 DEG C, to be soaked 15 minutes with 1% liquor potassic permanganate after cooling or formalin 100 times of immersions steep 1 hour naturally, soaks 6 hours-8 hours again after clear water is clean with clear water, then clean 2 times-3 times, to remove the surface of the seed thimerosal and its mucus of germinateing can be suppressed, dry moisture, clean the surface of the seed with clean cloth, 28 DEG C-30 DEG C constant temperature vernalization, sow during the long 2mm of bud;
B. sow, adopt program request; Down, about degree of depth 1.5cm is sowed at seedling culture hole plate to seed bud, and direction is consistent, and above covers the blinding of thick layer 1cm-1.5cm, irrigates the water containing bactericide, neatly puts in seedbed, and different cultivars and material do different marks;
C. nursery, before stock is emerged, seedbed day temperature remains on 25 DEG C-28 DEG C, and evening, temperature remained on 15 DEG C-18 DEG C, after emerging, and daytime, canopy temperature control was below 25 DEG C, remained on 15 DEG C-18 DEG C evening; After the seed of more than 50% breaks ground, throw off plastic film; After young plant all sprouts, control water, watering will " sees dry see wet ", and namely morning, matrix was watered, and time at dusk, matrix is shown in dry, and the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon can be used for grafting with the true leaf of five points of coin-size.
During described grafting is cultivated, temperature, humidity, illumination condition are as follows:
A. temperature, needs 2 days-4 days, keeps day temperature 26 DEG C-28 DEG C, night temperatures 22 DEG C-24 DEG C this period from grafting to wound healing, and temperature is higher than necessary aeration-cooling when 32 DEG C; Reduce temperature after wound healing, maintain 23 DEG C-25 DEG C daytime, maintain 20 DEG C-22 DEG C in the evening; Normal management in seedling stage is entered after 8-10 days;
B. humidity, first 2 days humidity requirements more than 95% after grafting, if humidity is too low, adopts water spray or watering earthward, directly can not spray water in the strain of wound healing forward direction grafting; Sprayable humidification after wound healing, but blade face can not ponding; Along with ventilation time extends, humidity is progressively reduced to 85%;
C. illumination, shading is cultivated and is sooner or later seen natural daylight afterwards in 3 days, progressively increases light transmittance afterwards, extends the printing opacity time, remove shade or net completely after 7 days-10 days;
D.14 the sprout tillers of stock is removed in sky later.
The present invention compared with prior art, has the following advantages and effect:
1, the sharpest edges of Plant Tissue Breeding are can obtain a large amount of clone plant fast at short notice, genetic stability is high, the present invention adopts watermelon plantlet in vitro to be scion, scion output can reach dozens or even hundreds of times of the scion production method of tradition " seed one seedling ", for the improved seeds seedling of seeding costliness, cost can reduce greatly.
2, this invention technology is compared with routine techniques, and the not underproduction, does not reduce graft survival rate.
3, this invention technology is compared with routine techniques, and with short production cycle, reproduction coefficient is large.
4, this invention can obtain low cost, the remarkable result of high profit.
Embodiment
With commodity kind aspermous watermelon plantlet in vitro for scion and cucurbit stock carry out grafting for example, scion hardening, stock prepare and grafting procedures is all carried out in green house, specifically implement:
A grafting method for watermelon plantlet in vitro, the steps include:
A, stock prepare, and stock seed is seeded in nutritive cube through vernalization, are used for grafting during the true leaf of the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon with five points of coin-size;
B, scion prepare, and growth medium grows to 25 days-30 days, grows to the plantlet in vitro of 2cm-3cm, and hardening of uncapping in 20 DEG C of-25 DEG C of greenhouse by solar heats is after 2 days-4 days, select that growing way is consistent, the plantlet in vitro of band 4 blades does scion;
C, the grafting of tip cutting method, first erase rootstock seedling the 1st true leaf base portion during grafting together with growing point, then with through 1%KnMO
4the tip of the bamboo let of solution disinfection is from growing point position to another cotyledon petiole base portion oblique cutting at 45 °, scion seedling is at distance plantlet in vitro foot blade 1cm place blade wedging, entered by the scion oblique cutting cut in the jack that bamboo let on stock puts on, stock is exposed at tip;
D, grafting are cultivated, grafting heat and moisture preserving, and seedling dish plastic film wraps up, and shading in first 3 days is cultivated, and throws off plastic film after 7 days, within 14 days, remove the sprout tillers of stock later;
E, grafting are transplanted, and grafting grafting tool two leaf of 22 days-28 days wholeheartedly can transplant land for growing field crops.
Described stock is cucurbit or pumpkin seedling.
Described stock prepares, and comprises the steps:
A. presoaking and germinating, with the water seed soaking of 55 DEG C, continuous stirring, until water temperature drops to 30 DEG C, to be soaked 15 minutes with 1% liquor potassic permanganate after cooling or formalin 100 times of immersions steep 1 hour naturally, soaks 6 hours-8 hours again after clear water is clean with clear water, then clean 2 times-3 times, to remove the surface of the seed thimerosal and its mucus of germinateing can be suppressed, dry moisture, clean the surface of the seed with clean cloth, 28 DEG C-30 DEG C constant temperature vernalization, sow during the long 2mm of bud;
B. sow, adopt program request; Down, about degree of depth 1.5cm is sowed at seedling culture hole plate to seed bud, and direction is consistent, and above covers the blinding of thick layer 1cm-1.5cm, irrigates the water containing bactericide, neatly puts in seedbed, and different cultivars and material do different marks;
C. nursery, before stock is emerged, seedbed day temperature remains on 25 DEG C-28 DEG C, and evening, temperature remained on 15 DEG C-18 DEG C, after emerging, and daytime, canopy temperature control was below 25 DEG C, remained on 15 DEG C-18 DEG C evening; After the seed of more than 50% breaks ground, throw off plastic film; After young plant all sprouts, control water, watering will " sees dry see wet ", and namely morning, matrix was watered, and time at dusk, matrix is shown in dry, and the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon can be used for grafting with the true leaf of five points of coin-size.
During described grafting is cultivated, temperature, humidity, illumination condition are as follows:
A. temperature, needs 2 days-4 days, keeps day temperature 26 DEG C-28 DEG C, night temperatures 22 DEG C-24 DEG C this period from grafting to wound healing, and temperature is higher than necessary aeration-cooling when 32 DEG C; Reduce temperature after wound healing, maintain 23 DEG C-25 DEG C daytime, maintain 20 DEG C-22 DEG C in the evening; Normal management in seedling stage is entered after 8-10 days;
B. humidity, first 2 days humidity requirements more than 95% after grafting, if humidity is too low, adopts water spray or watering earthward, directly can not spray water in the strain of wound healing forward direction grafting; Sprayable humidification after wound healing, but blade face can not ponding; Along with ventilation time extends, humidity is progressively reduced to 85%;
C. illumination, shading is cultivated and is sooner or later seen natural daylight afterwards in 3 days, progressively increases light transmittance afterwards, extends the printing opacity time, remove shade or net completely after 7 days-10 days;
D.14 the sprout tillers of stock is removed in sky later.
Claims (1)
1. a grafting method for watermelon plantlet in vitro, the steps include:
A, stock prepare, and stock seed is seeded in seedling culture hole plate through vernalization, are used for grafting during the true leaf of the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon with five points of coin-size;
B, scion prepare, and growth medium grows to 25 days-30 days, grows to the plantlet in vitro of 2cm-3cm, and hardening of uncapping in 20 DEG C of-25 DEG C of greenhouse by solar heats is after 2 days-4 days, select that growing way is consistent, the plantlet in vitro of band 4 blades does scion;
C, the grafting of tip cutting method, first erase rootstock seedling the 1st true leaf base portion during grafting together with growing point, then with through 1%KMnO
4the tip of the bamboo let of solution disinfection is from growing point position to another cotyledon petiole base portion oblique cutting at 45 °, scion seedling is at distance plantlet in vitro foot blade 1cm place blade wedging, entered by the scion oblique cutting cut in the jack that bamboo let on stock puts on, stock is exposed at tip;
D, grafting are cultivated, grafting heat and moisture preserving, and seedling dish plastic film wraps up, and shading in first 3 days is cultivated, and throws off plastic film after 7 days, within 14 days, remove the sprout tillers of stock later;
E, grafting are transplanted, and grafting grafting tool two leaf of 22 days-28 days wholeheartedly can transplant land for growing field crops;
Described stock is cucurbit or pumpkin seedling;
Described stock prepares, and comprises the steps:
A. presoaking and germinating, with the water seed soaking of 55 DEG C, continuous stirring, until water temperature drops to 30 DEG C, to be soaked 15 minutes with 1% liquor potassic permanganate after cooling or formalin 100 times of immersions steep 1 hour naturally, soaks 6 hours-8 hours again after clear water is clean with clear water, then clean 2 times-3 times, to remove the surface of the seed thimerosal and its mucus of germinateing can be suppressed, dry moisture, clean the surface of the seed with clean cloth, 28 DEG C-30 DEG C constant temperature vernalization, sow during the long 2mm of bud;
B. sow, adopt program request; Down, degree of depth 1.5cm is sowed at seedling culture hole plate to seed bud, and direction is consistent, and above covers the blinding of thick layer 1cm-1.5cm, irrigates the water containing bactericide, neatly puts in seedbed, and different cultivars and material do different marks;
C. nursery, before stock is emerged, seedbed day temperature remains on 25 DEG C-28 DEG C, and evening, temperature remained on 15 DEG C-18 DEG C, after emerging, and daytime, canopy temperature control was below 25 DEG C, remained on 15 DEG C-18 DEG C evening; After the seed of more than 50% breaks ground, throw off plastic film; After young plant all sprouts, control water, watering will " sees dry see wet ", and namely morning, matrix was watered, and time at dusk, matrix is shown in dry, and the open and flat and tool a slice of stock seedling age 15 days-20 days, two panels cotyledon can be used for grafting with the true leaf of five points of coin-size;
During described grafting is cultivated, temperature, humidity, illumination condition are as follows:
A. temperature, needs 2 days-4 days, keeps day temperature 26 DEG C-28 DEG C, night temperatures 22 DEG C-24 DEG C this period from grafting to wound healing, and temperature is higher than necessary aeration-cooling when 32 DEG C; Reduce temperature after wound healing, maintain 23 DEG C-25 DEG C daytime, maintain 20 DEG C-22 DEG C in the evening; Normal management in seedling stage is entered after 8-10 days;
B. humidity, first 2 days humidity requirements more than 95% after grafting, if humidity is too low, adopts water spray or watering earthward, directly can not spray water in the strain of wound healing forward direction grafting; Spray humidification after wound healing, but blade face can not ponding; Along with ventilation time extends, humidity is progressively reduced to 85%;
C. illumination, shading is cultivated and is sooner or later seen natural daylight afterwards in 3 days, progressively increases light transmittance afterwards, extends the printing opacity time, remove shade or net completely after 7 days-10 days;
D.14 the sprout tillers of stock is removed in sky later.
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Families Citing this family (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104106381A (en) * | 2014-07-02 | 2014-10-22 | 徐跃成 | Secondary grafting planting method for watermelons |
| CN104604541A (en) * | 2015-02-27 | 2015-05-13 | 刘伟 | Watermelon grafting seedling-cultivating method |
| CN106069218B (en) * | 2016-06-28 | 2019-03-29 | 江西武功鑫强生态农业开发有限公司 | A method of improving batch production greenhouse watermelon seedling graft seedling growth survival rate |
| CN106358770A (en) * | 2016-08-31 | 2017-02-01 | 宿州市埇桥区九里梦种植专业合作社 | Method for raising watermelon seedlings by grafting |
| CN106305159A (en) * | 2016-08-31 | 2017-01-11 | 宿州市埇桥区九里梦种植专业合作社 | Method for increasing grafting survival rate of Jianghuai Hami melons |
| CN107197698A (en) * | 2017-06-21 | 2017-09-26 | 陈付记 | A kind of root pruning and grafting method for culturing seedlings that watermelon is cultivated for second crop soil |
| CN107211700A (en) * | 2017-06-29 | 2017-09-29 | 安徽信福乡田生态农业有限公司 | A kind of nuisanceless watermelon breeding method |
| CN107466648A (en) * | 2017-08-31 | 2017-12-15 | 宁涛 | A kind of summer and autumn sunshade Pregermination and seedling breeding method of greenhouse watermelon |
| CN108575364A (en) * | 2018-04-02 | 2018-09-28 | 山东伟丽种苗有限公司 | Two points one line graftings |
| CN109673290B (en) * | 2019-01-14 | 2021-02-26 | 华中农业大学 | Watermelon top-inserting grafting method for reducing rootstock sprouting regeneration |
| CN113796219A (en) * | 2021-09-06 | 2021-12-17 | 河北农业大学 | Watermelon tissue culture seedling out-of-tube grafting and transplanting method |
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| CN101731089B (en) * | 2009-12-04 | 2011-11-30 | 武汉市农业科学研究所 | Method for producing seedless watermelon seedlings |
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Address after: 430345 Hubei Province, Wuhan city Huangpi District Wu lake agricultural agricultural ecological park in Wuhan city of Wuhan City Patentee after: INSTITUTE OF CROP SCIENCES OF WAAST Address before: 430345 Hubei Province, Wuhan city Huangpi District Wu lake agricultural agricultural ecological park in Wuhan city of Wuhan City Patentee before: Wuhan Institute of Agricultural Science |
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