CN110574684A - rapid breeding method of longya lily bulbs - Google Patents
rapid breeding method of longya lily bulbs Download PDFInfo
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- CN110574684A CN110574684A CN201910947711.2A CN201910947711A CN110574684A CN 110574684 A CN110574684 A CN 110574684A CN 201910947711 A CN201910947711 A CN 201910947711A CN 110574684 A CN110574684 A CN 110574684A
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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Abstract
the invention relates to the technical field of seed breeding, and discloses a rapid breeding method of lily bulb, which comprises the steps of processing and sterilizing an explant, inducing adventitious buds of lily bulb, proliferating and culturing lily bulb and expanding and rooting lily bulb. The invention has good disinfection effect, greatly reduces pollution rate, adopts natural illumination for culture, does not need to turn on a culture lamp, saves the cost of the culture process, utilizes the tender stem as an ectosome for quick induction, utilizes the corm for breeding the corm for quick proliferation, adopts the measure of accelerating the expansion of the corm, and achieves the purpose of quick speed of the whole breeding system; most of viruses are removed from the seedball bred by the method, the seed production time and period are shortened, the seedball obtained by the expanding rooting treatment of the method is large in size and strong in root system, and finally the obtained germplasm material is good in quality.
Description
Technical Field
The invention relates to the technical field of seed breeding, in particular to a rapid breeding method of lilium brownii seedballs.
Background
The longya lily is one of three main cultivated varieties in China, is long-standing, is a famous and special-quality variety in edible, medicinal and ornamental lily, and is also a variety with the best nutritive value. The bulb is spherical, the transverse diameter is 2-2.4 cm, and the scales are needle-shaped and white and are yellowish-white or slightly pink when exposed to the sun. The high content of starch, rich nutrition, light and bitter taste, the height of overground stems is about 90-120 cm, the leaves are scattered, and the shape of the inverted needles is adopted. The flower is milky and fragrant, and its bulb can be used as both food and medicine. According to records of medical records of Jinkui Yao, Ben Cao gang mu and Chinese pharmacopoeia, lily has the effects of moistening lung, relieving cough, calming heart, tranquilizing mind, regulating spleen, invigorating stomach, strengthening middle-jiao, replenishing qi, facilitating urination and defecation, relieving innominate toxic swelling, stopping bleeding and the like. Modern medical research proves that colchicine, colchicine and the like contained in lily have obvious inhibiting effect on mitosis of human cells, and have obvious curative effect on inhibiting cancer cells, acute gout and the like. Bulbus Lilii can also be used for extracting glucose and quinine. The lily bulb is large in bulb size, long in sheet, thick in meat, solid, white in color, delicious in taste, rich in nutrition and obvious in drug effect, and detected by a detection station in Jiangxi, each hundred grams of lily bulb contains 65% of water, 23.8% of carbohydrate and 5.9% of total dietary fiber, wherein pectin is 4.8%, protein is 4%, fat is 0.1%, water is 1.1%, calcium is 0.9%, iron is 0.09%, and heat is 132 kcal. Therefore, the lily is a green health food integrating food and medicine.
at present, the longya lily scales are the most commonly used tissue culture materials for rapid propagation of the longya lily, the longya lily scales are usually used for inducing adventitious buds and then carrying out proliferation on the adventitious buds, the longya lily scales are seriously infected with bacteria, the disinfection is often incomplete, the repeated pollution phenomenon in the culture process is very serious, the seed production time of the longya lily is prolonged while the workload is increased, the regeneration efficiency is not ideal, and the quality of the finally obtained germplasm materials for propagation is not ideal.
Disclosure of Invention
Based on the problems, the invention provides the rapid breeding method of the longya lily seedball, which can shorten the seed production time and period, has good detoxification effect and good quality of germplasm materials.
In order to solve the technical problems, the invention provides a rapid breeding method of the longya lily seedball, which comprises the following steps:
s1: treatment and disinfection of explants
Selecting fresh tender stem of Lilium Candidum, removing leaves, soaking in 75% ethanol for 30s, soaking in 0.2% mercuric chloride for 10-15min, washing with sterile water for 5 times, and air drying;
s2: induction of adventitious bud of longya lily
Cutting the tender stem of the lilium brownii processed in the step S1 into stem segments with the length of 1cm on a super clean bench, inoculating the stem segments on an induction culture medium, culturing at the temperature of 25 ℃, culturing by natural illumination, and differentiating a plurality of adventitious buds of the lilium brownii from each segment of tender stem after 15-20 days;
S3: proliferation culture of lily bulb
Inoculating the lily dragon 'S lily bulbs induced in the step S2 on a proliferation culture medium for culturing at 25 ℃, adopting natural illumination for culturing, and after 35-40 days, separating 3-4 proliferation bulbs from a single lily dragon' S lily bulb; then repeating the proliferation culture of the bulbodium for 30 days each time;
S4: expansion and rooting of proliferation bulblet of Lilium Candidum
And (4) inoculating the proliferated bulblets obtained in the step (S3) on an expanded rooting culture medium, placing the proliferated bulblets in a dark environment for culture at the culture temperature of 25 ℃, directly planting the expanded rooting bulblets in a field after 2 months of culture, and obtaining the longya lily bulbs after one year of planting.
further, the induction medium in step S2 is an improved MS medium, specifically, 1.5-3 mg/L6-BA, 0.05mg/L LNAA, 45g/L sucrose and 5g/L agar powder are added to the MS medium, and the PH value is 5.8.
Further, the enrichment medium in step S3 is an improved MS medium, specifically, 0.5 mg/L6-BA, 0.05mg/L LNAA, 45g/L sucrose, and 5g/L agar powder are added to the MS medium, and the PH value is 5.8.
Further, the expanded rooting medium in step S4 is an improved MS medium, specifically, the MS medium is supplemented with 0.01mg/LNAA, 8mg/L monopotassium phosphate, 3g/L activated carbon, 60g/L sucrose, and 5g/L agar powder, and the PH is 5.8, and the medium is cultured in the dark.
Further, the diameter of the bulb with expanded roots selected after 2 months of culture in the step S4 is 1.5-2cm, and the root system is strong.
Compared with the prior art, the invention has the beneficial effects that: the invention has good disinfection effect, greatly reduces pollution rate, adopts natural illumination for culture, does not need to turn on a culture lamp, saves the cost of the culture process, utilizes the tender stem as an ectosome for rapid induction, utilizes the corm for breeding the corm for rapid proliferation, adopts the measure of accelerating the expansion of the corm, and achieves the purpose of rapid speed of the whole breeding system. Most of viruses are removed from the seedball bred by the method, the seed production time and period are shortened, the seedball obtained by the expanding rooting treatment of the method is large in size and strong in root system, and finally the obtained germplasm material is good in quality.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to examples, and the exemplary embodiments and descriptions thereof are only used for explaining the present invention and are not used as limitations of the present invention.
example (b):
The rapid breeding method of the lilium brownii seedball comprises the following steps:
S1: treatment and disinfection of explants
selecting fresh tender stems of the lilium brownii after refrigeration or tender stems which grow from the ground in the spring of the next year after planting, peeling off leaves, soaking the tender stems in 75% alcohol for 30s on a super clean workbench, then soaking the tender stems in 0.2% mercuric chloride for 10-15min, then washing the tender stems with sterile water for 5 times, and placing the tender stems on the workbench for airing for later use; in this embodiment, the soaking time with 0.2% mercuric chloride is 12 min; the pollution rate is not more than 5 percent;
S2: induction of adventitious bud of longya lily
Cutting the tender stem of the lilium brownii processed in the step S1 into stem segments with the length of 1cm on a super clean bench, inoculating the stem segments on an induction culture medium, culturing at the temperature of 25 ℃, culturing by natural illumination, and differentiating dozens of adventitious buds of the lilium brownii from each segment of tender stem after 15-20 days; the induction culture medium is an improved MS culture medium, 1.5-3 mg/L6-BA, 0.05mg/LNAA, 45g/L sucrose and 5g/L agar powder are added to the MS culture medium, the pH value is 5.8, the 6-BA is 6-benzylaminopurine, NAA is naphthylacetic acid, the induction time is short by using the tender stem of the lilium brownii as an explant, the induction differentiation rate is extremely high, the proliferation female parent can be quickly obtained, and the breeding time is greatly shortened;
S3: proliferation culture of lily bulb
Inoculating the adventitious buds of the lily bulb of longya obtained by induction in the step S2 on a proliferation culture medium for culture at the culture temperature of 25 ℃, adopting natural illumination to culture for 35-40 days until 3-4 bulblets are differentiated from a single lily bulb of longya, and then utilizing the bulblets to repeat the proliferation culture for 30 days; in the embodiment, the adventitious buds are cultured for 40 days in a proliferation manner, wherein the proliferation culture medium is an improved MS culture medium, specifically, 0.5 mg/L6-BA, 0.05mg/L LNAA, 45g/L sucrose and 5g/L agar powder are added to the MS culture medium, the pH value is 5.8, the purpose of utilizing the corm to breed the corm and increasing the corm by more than 3 times is achieved, the concentration of 6-BA is reduced in the formula of the MS culture medium, and the corm is favorable for forming a compact sphere;
S4: expansion and rooting of proliferation bulblet of Lilium Candidum
inoculating the proliferated bulblets obtained in the step S3 on an expanded rooting culture medium, placing the proliferated bulblets in a dark environment for culture at the culture temperature of 25 ℃, directly planting the expanded rooting bulblets in a field after culturing for 2 months, wherein the diameter of the selected expanded rooting bulblets is 1.5-2cm, the root system of the bulblets is robust, and the longya lily bulbs can be obtained after being planted for one year; the expanding rooting culture medium is an improved MS culture medium, specifically, the MS culture medium is added with 0.01mg/LNAA, 8mg/L monopotassium phosphate, 3g/L active carbon, 60g/L sucrose and 5g/L agar powder, the pH value is 5.8, and the culture is carried out in the dark, and the monopotassium phosphate is added into the formula of the MS culture medium and the culture is carried out in the dark, so that the expansion of the bulb is accelerated.
the above is an embodiment of the present invention. The embodiments and specific parameters in the embodiments are only for the purpose of clearly illustrating the process of verifying the invention and are not intended to limit the scope of the invention, which is defined by the claims, and all the equivalent structural changes made by applying the content of the specification of the invention should be covered by the scope of the invention.
Claims (5)
1. The rapid breeding method of the lilium brownii seedball is characterized by comprising the following steps:
s1: treatment and disinfection of explants
Selecting fresh tender stem of Lilium Candidum, removing leaves, soaking in 75% ethanol for 30s, soaking in 0.2% mercuric chloride for 10-15min, washing with sterile water for 5 times, and air drying;
S2: induction of adventitious bud of longya lily
Cutting the tender stem of the lilium brownii processed in the step S1 into stem segments with the length of 1cm on a super clean bench, inoculating the stem segments on an induction culture medium, culturing at the temperature of 25 ℃, culturing by natural illumination, and differentiating a plurality of adventitious buds of the lilium brownii from each segment of tender stem after 15-20 days;
S3: proliferation culture of lily bulb
inoculating the lily dragon 'S lily bulbs induced in the step S2 on a proliferation culture medium for culturing at 25 ℃, adopting natural illumination for culturing, and after 35-40 days, separating 3-4 proliferation bulbs from a single lily dragon' S lily bulb; then repeating the proliferation culture of the bulbodium for 30 days each time;
S4: expansion and rooting of proliferation bulblet of Lilium Candidum
And (4) inoculating the proliferated bulblets obtained in the step (S3) on an expanded rooting culture medium, placing the proliferated bulblets in a dark environment for culture at the culture temperature of 25 ℃, directly planting the expanded rooting bulblets in a field after 2 months of culture, and obtaining the longya lily bulbs after one year of planting.
2. The rapid propagation method of lily bulbs of Longya Korea as claimed in claim 1, wherein the inducing medium in step S2 is an improved MS medium, specifically, the MS medium is supplemented with 1.5-3 mg/L6-BA, 0.05mg/L NAA, 45g/L sucrose and 5g/L agar powder, and the pH value is 5.8.
3. The rapid propagation method of lily bulbs of the longya variety as claimed in claim 1, wherein the enrichment medium in step S3 is an improved MS medium, specifically, 0.5 mg/L6-BA, 0.05mg/LNAA, 45g/L sucrose and 5g/L agar powder are added to the MS medium, and the PH value is 5.8.
4. The rapid propagation method of lily bulbs of Longya hybrida according to claim 1, wherein the expanded rooting medium in step S4 is an improved MS medium, specifically, the MS medium is supplemented with 0.01mg/LNAA, 8mg/L potassium dihydrogen phosphate, 3g/L activated carbon, 60g/L sucrose and 5g/L agar powder, the pH value is 5.8, and the bulbs are cultured in the dark.
5. The rapid propagation method of lily bulbs of longya according to claim 1, wherein the diameter of the bulb of enlarged roots selected after 2 months of cultivation in step S4 is 1.5-2cm, and the root system is robust.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112673956A (en) * | 2020-12-18 | 2021-04-20 | 湖北顗泉种业生物技术有限公司 | Method for greatly improving variety and quality of longya lily |
Citations (5)
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| WO1991014357A1 (en) * | 1990-03-28 | 1991-10-03 | THE UNITED STATE OF AMERICA, represented by THE SECRETARY, UNITED STATES DEPARTMENT OF COMMERCE | A METHOD FOR PRODUCING $i(LILIUM ELEGANS) |
| CN1903017A (en) * | 2005-07-29 | 2007-01-31 | 潘利军 | Method for separating bulbs of lily tissue culture |
| CN102440185A (en) * | 2010-10-14 | 2012-05-09 | 赵祥云 | Fast culturing method for lily virus-free seed ball |
| CN105210877A (en) * | 2015-10-20 | 2016-01-06 | 韦丽 | A kind of Lilium brownii var viridulum method for quickly breeding |
| CN109463282A (en) * | 2018-12-14 | 2019-03-15 | 河南城建学院 | A kind of Lilium brownii var viridulum clove numerous rooting method fastly |
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2019
- 2019-10-08 CN CN201910947711.2A patent/CN110574684A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO1991014357A1 (en) * | 1990-03-28 | 1991-10-03 | THE UNITED STATE OF AMERICA, represented by THE SECRETARY, UNITED STATES DEPARTMENT OF COMMERCE | A METHOD FOR PRODUCING $i(LILIUM ELEGANS) |
| CN1903017A (en) * | 2005-07-29 | 2007-01-31 | 潘利军 | Method for separating bulbs of lily tissue culture |
| CN102440185A (en) * | 2010-10-14 | 2012-05-09 | 赵祥云 | Fast culturing method for lily virus-free seed ball |
| CN105210877A (en) * | 2015-10-20 | 2016-01-06 | 韦丽 | A kind of Lilium brownii var viridulum method for quickly breeding |
| CN109463282A (en) * | 2018-12-14 | 2019-03-15 | 河南城建学院 | A kind of Lilium brownii var viridulum clove numerous rooting method fastly |
Non-Patent Citations (4)
| Title |
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| BONG HEE HAN等: "In vitro micropropagation of Lilium longiflorum ‘Georgia’ by shoot formation as influenced by addition of liquid medium", 《SCIENTIA HORTICULTURAE》 * |
| 徐洪星: "大理百合组织培养和快速繁育研究", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
| 王亚等: "‘西伯利亚’百合组培快繁体系的优化", 《中国园艺学会观赏园艺专业委员会2012年学术年会》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112673956A (en) * | 2020-12-18 | 2021-04-20 | 湖北顗泉种业生物技术有限公司 | Method for greatly improving variety and quality of longya lily |
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