CN112673956A - Method for greatly improving variety and quality of longya lily - Google Patents
Method for greatly improving variety and quality of longya lily Download PDFInfo
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Abstract
A method for greatly improving variety and quality of Lilium Brownii comprises selecting field Lilium Brownii variety, cleaning bulb fruit, sterilizing and detoxicating scale, and preparing yellow matrix explant; inoculating the explant to a bulblet induction culture medium; inducing bulblet explants and bulblet optimization; transferring the bulblets into a bulblet multiplication culture medium for multiplication, and then transferring into a differentiation culture medium for differentiation; inoculating the differentiated seedling to a strong seedling rooting culture medium; hardening the rooted seedlings, and cultivating the hardened rooted seedlings into virus-free bulb first-generation fruit seedlings in a seedling cultivation greenhouse; edible lily bulb corms (commercial fruits) are planted in the open air in the field. The method improves the variety and the quality of the lilium brownii by using a modern biotechnology method, the acre yield of the cultivated detoxified first-generation fruit seedlings is improved by more than 50 percent compared with the acre yield of the conventional breeding and planting method after the field open planting for one year, the bulb and the bulb of the lilium brownii have better appearance, no or less jute spots, no or less plant diseases and insect pests and low pesticide residue.
Description
Technical Field
The invention relates to the field of biotechnology and a field variety optimization method, in particular to a method for greatly improving varieties of longya lilies and improving the quality of the varieties of the longya lilies.
Background
The longya lily is one of three lily varieties in China, is a perennial herb plant in Liliaceae, prefers a half-shady environment, and is one of cold-resistant and heat-resistant lily varieties. The soil conditioner is suitable for growing in sandy loam with deep soil layer, good drainage and loose and fertile soil, prefers slightly acidic soil and is also suitable for light lime soil. The bulb of the lilium brownii is spherical, the diameter is about 2-4.5 cm, and the scales are needle-shaped and white. High starch content, rich nutrients, light taste and no bitter taste. The stem is about 100 cm high, the leaves are scattered, and the shape of the inverted needle is changed into the inverted egg shape; the flower is milky and fragrant, and can be used for producing seeds; the flowering period is 5-6 months, and the fruit ripening period is 9-10 months. The subterranean bulb of lily is rich in starch, protein, fat and various vitamins and minerals required by the human body. 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 is a traditional product, has rich nutrition and obvious drug effect, and contains 65 percent of water, 23.8 percent of carbohydrate and 5.9 percent of total dietary fiber in each hundred grams of lily. The Longya lily is a green health food integrating food and medicine.
The field planting of the longya lily is long-lived, and the history of the longya lily exceeds 1400 years. However, in the conventional breeding and planting methods such as bulb separation and scale embedding in the field, after multi-generation planting, the variety property is seriously degraded, and the yield of the bulb fruits is obviously reduced. In addition, in the traditional seed reserving planting method, harmful bacteria and viruses are accumulated in soil and bulbus lilii, bacterial infection and plant diseases and insect pests are increasingly serious, the disease resistance of the plant is weak, a large amount of pesticide needs to be sprayed in the planting process, and the soil and the lily bulbus lilii have more pesticide residues, so that the human health is influenced.
Disclosure of Invention
The invention provides a method for greatly improving variety and quality of lilium brownii, which can solve the problems of variety degeneration, bacterial infection and serious pest and disease damage of lilium brownii, greatly improve acre yield and quality, reduce pest and disease damage, reduce the use amount of pesticides and provide high-quality and high-yield seedlings for lily planting industry.
In order to achieve the above purpose, the solution of the invention is as follows:
a method for greatly improving variety and quality of Longya lily is characterized by comprising the following steps:
firstly, the variety of the field planted Longya lily is optimized
Selecting the bulb cones with regular appearance, smooth and white scales, flat mouth shape formed by tightly wrapping the scales on the dragon teeth and attaching the scales to the upper end core mouth, dense bulb cones, large lumps, single fruit shape, thick and thick scales, no jute spots or less jute spots; planting the bulb fruit seedlings or the stock seed scales bred according to the standard in a seed source area, reserving a robust stock plant, and selecting biennial bulb fruit of the stock plant as stock seed bulb fruit for tissue culture and rapid propagation;
washing bulb and peeling, sterilizing and detoxicating scale
Cleaning and removing mud from the bulb of the preferred stock lily bulb, peeling off the scale at the central part, and cleaning, sterilizing and detoxifying the scale to obtain sterilized detoxified stock scale;
preparation of detoxicated yellow matrix explant
Absorbing water from the disinfected and detoxified scales to prepare detoxified yellow matrix explants;
fourth, induction and optimization of bulblet
Inoculating the yellow matrix explant obtained in the third step to a bulblet induction culture medium for induction bulblet culture, wherein the induction culture medium comprises the following components in percentage by weight: MS minimal medium, 6-BA 1.6mg/L-3.0mg/L, NAA 0.1.1-0.7 mg/L, KT 0.2.2 mg/L-2.0mg/L, pH value 5.8; the culture conditions were: after the small tender shoots grow out after the illumination intensity is 2000-; the light source is a plant lamp, the illumination time is 12h/d, the temperature is 24-28 ℃, and the green and robust bulblet can be induced from the explant;
from the explant in which the bulblet is induced, preferably the bulblet of the explant in which the 4-5 buds are induced enters the next cultivation;
proliferation of bulblet
Transferring the bulblets obtained in the fourth step to a bulblet multiplication culture medium, wherein the ratio of the multiplication culture medium is as follows: MS minimal medium, NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, white sugar 30g/L, pH value 5.8, the culture conditions at this stage are as follows: irradiating with plant lamp light source at illumination intensity of 3500 Lux 4500Lux for 12h/d at 24-28 deg.C, and performing secondary proliferation every 50-55 days;
differentiation of bulblet
Transferring the bulblets obtained in the fifth step into a differentiation medium for differentiation, wherein the differentiation medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, potato extract 60-100g/L, pH value 5.8; the culture conditions were: the illumination intensity is 4000-;
seven, strengthening and rooting of seedlings and hardening seedlings
Transferring the differentiated seedling obtained in the sixth step into a strong seedling rooting culture medium, wherein the strong seedling rooting culture medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, banana extract 100-: under the conditions of illumination by a plant lamp light source, illumination intensity of 4000-;
placing the obtained rooted seedlings and the culture bottle in a ventilated greenhouse with illumination intensity of 7000-8000Lux and temperature of 15-28 ℃ for hardening seedlings for 8-15 days;
eighthly, cultivating the first-generation virus-free lily bulb seedlings in a seedling growing greenhouse
Taking out the acclimatized rooted seedlings from the bottle, washing the culture medium with tap water, and performing culture in a volume of 1: soaking in carbendazim solution of 1500 for 5 min, taking out, and filtering to dry;
planting the drained rooting seedlings in a seedling raising greenhouse in sandy loam, wherein the planting density per square meter is about 160-;
after one year, each rooted seedling can grow a first generation fruit seedling of the detoxified longya lily bulb with the weight of about 45-60 g;
nine, bulb corm of virus-free longya lily planted in open air
And (4) after autumn comes up every year, planting the first generation of virus-free bulb seedlings obtained by cultivation in the eighth step in an open sandy loam field, performing normal management in the field at ordinary times, and harvesting virus-free lily bulb cones which are good in quality, free or less in jute spots and dragon tooth scales, flat in mouth shape and clean and white in crystal in the next 7-8 months.
Further, in the second step, the disinfected and sterilized scales are placed in constant-temperature sterile water of 40 ℃ to be soaked for 3 hours for detoxification treatment.
Further, in the second step, firstly washing away the soil on the surface of the original bulb fruit by using tap water, removing the surface layer of the cleaned bulb fruit to obtain central part of the scale, soaking the scale in saturated phosphorus-free washing powder water for 10 minutes, then washing the scale with tap water for 120 minutes, washing the scale with sterile water for 1 time, firstly disinfecting the scale with 75% alcohol on a super-clean workbench for 15 seconds, washing the scale with sterile water for 3 times, soaking the scale with 0.1-0.2% mercuric chloride solution for 10 minutes, then washing the scale with sterile water for 6 times, and finally performing detoxification treatment.
Further, when the bulblets are induced in the fourth step, small leaves can grow out of part of the robust bulblets, the small leaves are cut off, and the small leaves and the bulblets are transferred to the enrichment culture medium in the fifth step for enrichment culture.
Further, when the bulblets are propagated in the fifth step, small leaves can grow out of part of the robust bulblets, and when the small leaves are cut off to participate in the next round of bulblet propagation, the robust bulblets can also grow out of the lower parts of the leaves.
Further, in the eighth step, when the rooted seedlings are planted in sandy loam, the soil ploughing depth is about 20cm, and ridging, furrow digging and good drainage are required.
Further, in the ninth step, when the first-generation virus-free bulb seedlings are planted in an open sandy loam field, the soil ploughing depth is about 30cm, and suspension ploughing, soil breaking, ridging, ditching and good drainage are required.
Further, in the third step, after the water content of the disinfected and detoxified scales is filtered, the scales with complete appearance are selected on a clean bench and cut into strip-shaped blocks with the length and width of about 2cm and 1cm, the parts close to the bottom of the blocks with the yellow matrix are preferably sliced to prepare explants with the yellow matrix and common explants, the common explants are discarded, and the explants with the yellow matrix are reserved.
Further, in the eighth step, the rooted seedlings are irrigated with rooting water on the same day of planting; on the 1 st to 10 th days, the illumination condition is controlled to be 4000-6000 Lux; after 10-15 days, the greenhouse naturally transmits light in the morning and evening, the illumination intensity is controlled to be 6000-; after 15-20 days, the lamp was irradiated with natural light.
Further, in the ninth step, the planting amount of each mu of land is about 800 jin of detoxified first-generation fruit seedlings, and 3800 jin of detoxified lily bulb cones can be harvested in each mu of planted land in 7-8 months in the next year.
Aiming at the problems of variety degeneration, multiple accumulated bacteria and viruses, serious pest and disease damage and the like of the lilium fargesii, the tissue culture seedling culture method for optimizing the variety of the lilium fargesii planted in the field, washing lily bulbs, sterilizing and detoxifying scales and detoxifying and rejuvenating female parents to recover the excellent properties ensures the inheritance and stability of good genes of the lilium fargesii, improves the yield per mu by more than 50 percent, greatly reduces the pest and bacterial infection and ensures that the surface layer of lily bulb fruits has no or few jute spots. The invention adopts the methods of tissue culture seedling raising and industrial batch production, so that the achievement of the invention can be industrialized immediately, and high-quality and high-yield seedlings are provided for the lily planting industry.
Detailed Description
In order to further explain the technical solution of the present invention, the present invention is explained in detail by the following specific examples.
The invention adopts the field longya lily bulb corm variety optimization and biotechnology method to solve the problems of variety degeneration, yield reduction, bacteria and virus accumulation and serious pest and disease damage of longya lily after multi-generation separate planting, the variety of the longya lily and high-quality genes are subjected to copy reduction, after variety optimization, bulb washing and stripping, scale disinfection and sterilization, tissue culture, seedling hardening, greenhouse seedling raising, the longya lily planted in the field by adopting the first generation of detoxified bulb seedlings or the second generation of bulb seedlings can be improved by more than 50 percent per mu, the bulb corm has few jute spots, and the appearance is more beautiful; the seed reserving and planting process within 4 generations of virus-free bulb fruit seedlings has no plant diseases and insect pests or greatly reduces the plant diseases and insect pests, and greatly reduces the using amount of pesticides in the planting process. The invention comprises the following steps:
first, variety optimization of the field planting of the lilium fargesii
Taking field divisions of planting areas such as Hunan Longhui, Longshan, Hubei Dangyang and the like as provenance protection fields, preferably selecting field-planted lily bulb cones as antenatal bulb cones with improved varieties according to eight standards of positive appearance, smooth and white scales, flat mouth shape formed by tightly wrapping the scales on dragon teeth and attaching the scales to the upper core mouth, dense bulb cones, large block heads, single fruit shape, thick and thick scales and no jute spots or few jute spots.
And (3) planting the bulb cones bred according to the standards or peeling scales thereof in a seed source region, eliminating weak and small plants, reserving strong stock plants, and performing the eight standards again to preferably select the bulb cones meeting the requirements from the bulb cones of the biennial stock plants as the protospecies bulb cones which are the original materials for tissue culture and rapid propagation.
In the embodiment, the perennial bulb cones are preferably selected from the fields of Hunan Longhui county, and the weight of a single fruit is between 300 and 400 g. And (3) separating the optimized bulb cones, planting the bulb cones in a seed source area of the Longhui county in Hunan province, and performing the standard again after planting for two years to optimize the bulb cones with the shapes of 1000 jin single fruits and the weight of more than 380 g, and transporting the bulb cones to a national forest fine variety breeding (Xiamen) demonstration base to be used as the original bulb cones for tissue culture.
Washing and flaking bulb cones, and sterilizing and detoxifying scales
The optimized protospecies bulb is cleaned, flaked, sterilized and detoxified according to the following steps. Washing the soil on the surface of the original bulb fruit with tap water, removing the surface scale of the cleaned bulb fruit to select the central scale, soaking the scale in saturated phosphorus-free washing powder water for 10 minutes, then washing with the tap water for 120 minutes, washing with sterile water for 1 time, disinfecting with 75% alcohol for 15 seconds, washing with the sterile water for 3 times, soaking with 0.1-0.2% mercuric chloride solution for 10 minutes, and then washing with the sterile water for 6 times on a super clean workbench; finally, placing the mixture in sterile water with constant temperature of 40 ℃ for soaking for 3h to obtain the tissue culture material of the virus-free scale.
In this embodiment, about 200 jin of the intermediate layer scales can be peeled off from 1000 jin of the lily bulb.
The explant subjected to heat treatment by constant-temperature sterile water soaking at 40 ℃ plays a role in detoxification, and can eliminate the explant with weak vitality and the offspring induced in the next step (because the cells with weak vitality and the explant are killed by scalding, the step plays a role in eliminating a part of non-robust explants), so that the explant with strong vitality and the induced offspring thereof can be obtained.
The following table 1 shows experimental data of tissue culture survival rate of induction of bulblets in a bulblet induction medium by constant-temperature sterile water soaking heat treatment at 40 ℃ and inoculation of non-treated explants, other tissue culture environments and treatment links are the same, and the experimental sample amount is respectively 100 explants.
Table 1: the explant is subjected to constant-temperature heat treatment at 40 ℃ and non-treatment tissue culture experiment observation data
Preparation of explant of three-yellow matrix
Draining water from the disinfected and detoxified scales, and cutting the scales with complete shapes into strip-shaped blocks with the length and width of about 2cm and 1cm on a super clean workbench; when cutting, the part with yellow matrix near the bottom is selected for preferential cutting. So as to prepare explants with the yellow matrix and general explants. The normal explants were discarded and explants with yellow stroma were left.
Fourth, induction and optimization of bulblet
Inoculating the detoxified yellow matrix explant obtained in the third step into a bulblet induction culture medium for induction culture, wherein the induction culture medium comprises the following components in percentage by weight: MS minimal medium, 6-BA (benzylpurine) 1.6mg/L-3.0mg/L, NAA (naphthalene acetic acid) 0.1-0.7mg/L, KT (kinetin) 0.2mg/L-2.0mg/L, pH value 5.8; the culture conditions were: under the conditions that the illumination intensity is 2000-; the light source is a plant lamp, and the temperature is 24-28 ℃. The explants are cultured in the induction medium for about 50-55 days to induce a green and robust bulblet.
Approximately 55% of explants were able to induce robust bulbs by the above method. And selecting the explants inducing the 4-5 bud thick bulblet from the explants inducing the bulblet to enter the next cultivation cycle, and discarding the explants inducing less than 3 buds and more than 5 buds. The proportion of explants with shoots 4-5 accounted for explants capable of inducing bulblet formation was about 60%.
When bulblet is induced, if the tissue culture experimental conditions are not well controlled, only callus can be induced, adventitious buds are induced through the callus, and bulblet is induced through the adventitious buds, so that the tissue culture efficiency and economic benefit are reduced seriously, and the quality of subsequent seedlings is influenced.
When the bulblets are induced in the fourth step, small leaves already grow on part of robust bulblets, and the small leaves are cut off to participate in the next step of bulblet proliferation, so that the proliferation efficiency is improved.
The method directly induces the thick and strong bulblets, cuts off the small leaves grown from a small part of the thick and strong bulblets in the next step for proliferation.
Proliferation of bulblet
Transferring the bulblets (and the small leaves) obtained in the fourth step into a bulblet multiplication culture medium, wherein the ratio of the multiplication culture medium is as follows: MS minimal medium (see table 3), NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, white sugar 30g/L, and pH value 5.8. The culture conditions at this stage were: the illumination intensity is about 3500 Lux 4500Lux, the illumination time is 12h/d, and the subculture proliferation can be carried out once every 50-55 days.
When the bulblets are proliferated in the fifth step, small leaves grow out of part of robust bulblets, and when the small leaves are cut off to participate in the next round of bulblet proliferation (namely, the bulblets are proliferated next time), the robust bulblets can also grow out of the lower parts of the leaves, so that the proliferation efficiency is greatly improved.
In this case, the small leaflets that proliferate in this step have small bulbs growing around their bottom.
The bulblets are respectively inoculated in MS and 1/2MS (macroelement is halved, and trace elements are unchanged) different basic proliferation culture media to carry out proliferation experiments, and the influence of different culture media on the bulblet proliferation effect is observed. Table 2 below shows experimental data on the effect of different basal media on the proliferation of bulblets, with the experimental sample size being 100 each.
The experiments show that different basic culture media have great influence on the proliferation rate of the bulblet. 1/2MS is used as a basic formula of MS basic culture medium after half of macroelements are reduced, and the element concentration requirement of the plant growth is lacked, so that the multiplication rate is not ideal in the culture process; the experiments show that the basic formula of the MS culture medium is more suitable for the multiplication culture of the bulblet of the lilium densefolium.
Table 2: influence of different basic culture media on proliferation rate of bulblet
Differentiation of bulblet
Transferring the bulblets obtained in the fifth step into a differentiation medium, wherein the differentiation medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, potato extract 60-100g/L, pH value 5.8. The culture conditions were: the illumination intensity is 4000-. After about 20 days, about 95% of the bulbs differentiated to give acceptable differentiated shoots.
Seven, strengthening and rooting of seedlings and hardening seedlings
Inoculating the differentiated seedling of the lilium fargesii obtained in the sixth step into a strong seedling rooting culture medium, wherein the strong seedling rooting culture medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, banana extract 100-. And culturing the differentiated seedling in the culture medium for about 15 days under the conditions of illumination of a plant lamp light source, illumination intensity of 4000-4500Lux, illumination time of 12h/d and temperature of 24-28 ℃, changing the illumination condition, adjusting the illumination intensity to 5500Lux-6000 Lux, illumination time of 12h/d, the light source of a plant lamp and the temperature of 24-28 ℃, and culturing for about 20 days to obtain the robust rooted seedling.
And placing the obtained rooted seedlings and culture bottles in a 15-28 ℃ ventilated greenhouse for hardening seedlings for 8-15 days, and then using the hardened seedlings for planting in a seedling raising greenhouse, wherein the illumination intensity is 7000-8000 Lux.
Eighthly, seedling raising greenhouse seedling raising
Taking the acclimatized rooted seedlings obtained in the seventh step out of the bottle, washing the culture medium with tap water, and performing culture in a volume of 1: soaking in carbendazim solution 1500 for 5 min, taking out, and filtering to dry.
Planting the drained rooting seedlings in a seedling raising greenhouse (film and sunshade net) of 8M by 50M sandy loam, wherein the planting density per square meter is about 160 and 180 rooting seedlings, the soil tilling depth is about 20cm, and ridging, ridge digging and ditch digging and good drainage are required.
Irrigating the rooted seedlings with rooting water on the same day as the planting of the rooted seedlings; on the 1 st to 10 th days, the illumination condition is controlled to be 4000-6000 Lux; after 10-15 days, the greenhouse naturally transmits light in the morning and evening, the illumination intensity is controlled to be 6000-; after 15-20 days, the lamp was irradiated with natural light.
After one year, one generation of fruit seedlings of the virus-free lily bulbs with weight of about 45-60 g can grow from each rooted seedling. The planting survival rate of the refined rooting seedlings is more than 95 percent.
Nine-day open planting
After autumn every year, planting the first generation of virus-free lily bulb seedlings obtained in the step eight in an open sandy loam field; planting prenatal preparation requirements: the land is cultivated in a suspension mode, soil is crushed, ridges are formed, the water is drained well, and the soil cultivation depth is about 30 cm; the planting amount of each mu of land is about 8000-. In 7-8 months of the next year, 5000 jin of detoxified lily bulb cones with good quality, no or few jute spots, flat shapes of dragon tooth scales and white crystal can be harvested in each mu of planting land.
The lily dragon tooth is detoxified and does not have plant diseases and insect pests after being planted for 6 years in Yanbian tests by using the method; the plant diseases and insect pests are greatly reduced when the seeds are tested in the Longhui county in Hunan province. The yield per mu is more than 50 percent of that of the traditional dragon tooth lily.
The method solves the problems of the traditional lily dragon tooth that the variety of the lily dragon tooth is degenerated, the accumulated bacteria and viruses are increased, the yield is reduced and the plant diseases and insect pests are serious. By means of great improvement on variety and quality of the lilium brownii and large-scale industrialized tissue culture seedling raising, large batches of excellent seedlings can be provided for farmers, and the acre yield is improved by more than 50% through experimental planting in Yanbian and Hunan Longhui county, the bulbar bulbs are smooth and white in crystal, fair in appearance and few in jute spots. The industrialization of the invention can create higher economic value and save land for growers.
The above embodiments are not intended to limit the form and style of the present invention, and any suitable changes or modifications made by those skilled in the art should be considered as not departing from the scope of the present invention.
Table 3: the ingredients of the MS medium are as follows
Claims (10)
1. A method for greatly improving variety and quality of Longya lily is characterized by comprising the following steps:
firstly, the variety of the field planted Longya lily is optimized
Selecting the bulb cones with regular appearance, smooth and white scales, flat mouth shape formed by tightly wrapping the scales on the dragon teeth and attaching the scales to the upper end core mouth, dense bulb cones, large lumps, single fruit shape, thick and thick scales, no jute spots or less jute spots; planting the bulb fruit seedlings or the stock seed scales bred according to the standard in a seed source area, reserving a robust stock plant, and selecting biennial bulb fruit of the stock plant as stock seed bulb fruit for tissue culture and rapid propagation;
washing bulb and peeling, sterilizing and detoxicating scale
Cleaning and removing mud from the bulb of the preferred stock lily bulb, peeling off the scale at the central part, and cleaning, sterilizing and detoxifying the scale to obtain sterilized detoxified stock scale;
preparation of detoxicated yellow matrix explant
Absorbing water from the disinfected and detoxified scales to prepare detoxified yellow matrix explants;
fourth, induction and optimization of bulblet
Inoculating the yellow matrix explant obtained in the third step to a bulblet induction culture medium for induction bulblet culture, wherein the induction culture medium comprises the following components in percentage by weight: MS minimal medium, 6-BA 1.6mg/L-3.0mg/L, NAA 0.1.1-0.7 mg/L, KT 0.2.2 mg/L-2.0mg/L, pH value 5.8; the culture conditions were: after the small tender shoots grow out after the illumination intensity is 2000-; the light source is a plant lamp, the illumination time is 12h/d, the temperature is 24-28 ℃, and the green and robust bulblet can be induced from the explant;
from the explant in which the bulblet is induced, preferably the bulblet of the explant in which the 4-5 buds are induced enters the next cultivation;
proliferation of bulblet
Transferring the bulblets obtained in the fourth step to a bulblet multiplication culture medium, wherein the ratio of the multiplication culture medium is as follows: MS minimal medium, NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, white sugar 30g/L, pH value 5.8, the culture conditions at this stage are as follows: irradiating with plant lamp light source at illumination intensity of 3500 Lux 4500Lux for 12h/d at 24-28 deg.C, and performing secondary proliferation every 50-55 days;
differentiation of bulblet
Transferring the bulblets obtained in the fifth step into a differentiation medium for differentiation, wherein the differentiation medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, 6-BA1.4-3.6mg/L, potato extract 60-100g/L, pH value 5.8; the culture conditions were: the illumination intensity is 4000-;
seven, strengthening and rooting of seedlings and hardening seedlings
Transferring the differentiated seedling obtained in the sixth step into a strong seedling rooting culture medium, wherein the strong seedling rooting culture medium comprises the following components in percentage by weight: MS minimal medium, NAA0.4-1.2mg/L, banana extract 100-: under the conditions of illumination by a plant lamp light source, illumination intensity of 4000-;
placing the obtained rooted seedlings and the culture bottle in a ventilated greenhouse with illumination intensity of 7000-8000Lux and temperature of 15-28 ℃ for hardening seedlings for 8-15 days;
eighthly, cultivating the first-generation virus-free lily bulb seedlings in a seedling growing greenhouse
Taking out the acclimatized rooted seedlings from the bottle, washing the culture medium with tap water, and performing culture in a volume of 1: soaking in carbendazim solution of 1500 for 5 min, taking out, and filtering to dry;
planting the drained rooting seedlings in a seedling raising greenhouse in sandy loam, wherein the planting density per square meter is about 160-;
after one year, each rooted seedling can grow a first generation fruit seedling of the detoxified longya lily bulb with the weight of about 45-60 g;
nine, bulb corm of virus-free longya lily planted in open air
And (4) after autumn comes up every year, planting the first generation of virus-free bulb seedlings obtained by cultivation in the eighth step in an open sandy loam field, performing normal management in the field at ordinary times, and harvesting virus-free lily bulb cones which are good in quality, free or less in jute spots and dragon tooth scales, flat in mouth shape and clean and white in crystal in the next 7-8 months.
2. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: and step two, soaking the disinfected and sterilized scales in constant-temperature sterile water of 40 ℃ for 3 hours for detoxification treatment.
3. The method for greatly improving variety and quality of longya lily according to claim 1 or 2, which is characterized in that: and in the second step, washing away soil on the surface of the original bulb fruit by using tap water, removing the surface layer of the washed bulb fruit to obtain central-part scales, soaking the scales in saturated phosphorus-free washing powder water for 10 minutes, then washing the scales with the tap water for 120 minutes, washing the scales with sterile water for 1 time, disinfecting the scales on a super-clean workbench for 15 seconds by using 75% alcohol, washing the scales with the sterile water for 3 times, soaking the scales in 0.1-0.2% mercuric chloride solution for 10 minutes, then washing the scales with the sterile water for 6 times, and finally performing detoxification treatment.
4. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: and step four, when the bulblets are induced, the small leaves can grow out from part of the robust bulblets, the small leaves are cut off, and the small leaves and the bulblets are transferred into the enrichment culture medium in the step five for enrichment culture.
5. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: and fifthly, when the bulblets are proliferated, small leaves can grow out from part of robust bulblets, and when the small leaves are cut off and participate in the next round of bulblet proliferation, the robust bulblets can also grow out from the lower parts of the leaves.
6. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: and step eight, when the rooted seedlings are planted in sandy loam, the soil ploughing depth is about 20cm, and ridging, furrow digging and good drainage are required.
7. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: in the ninth step, when the first-generation virus-free bulb seedlings are planted in an open sandy loam field, the soil ploughing depth is about 30cm, and suspension ploughing, soil crushing, ridging, ditching and good drainage are required.
8. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: in the third step, after the water content of the disinfected and detoxified scales is drained, the scales with complete shapes are selected on a super clean bench and cut into strip-shaped blocks with the length and width of about 2cm and 1cm, the parts close to the bottom of the blocks with the yellow matrix are preferentially sliced during the cutting to prepare explants with the yellow matrix and common explants, the common explants are discarded, and the explants with the yellow matrix are reserved.
9. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: in the eighth step, the rooted seedlings are irrigated with rooting water on the same day as the planting; on the 1 st to 10 th days, the illumination condition is controlled to be 4000-6000 Lux; after 10-15 days, the greenhouse naturally transmits light in the morning and evening, the illumination intensity is controlled to be 6000-; after 15-20 days, the lamp was irradiated with natural light.
10. The method for greatly improving variety and quality of longya lily as claimed in claim 1, which is characterized in that: in the ninth step, the planting amount of each mu of land is about 800 jin of detoxified first-generation fruit seedlings, and 3800 jin of detoxified lily bulb cones can be harvested in each mu of planted land in 7-8 months in the next year.
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101061790A (en) * | 2007-04-13 | 2007-10-31 | 浙江省农业科学院 | Quick virus-free group-cultivating and propagating method of OT type lily of hybrid group |
| CN109315288A (en) * | 2017-07-31 | 2019-02-12 | 赵义平 | A kind of tissue culture and rapid propagation method of lily |
| 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 |
| CN110574684A (en) * | 2019-10-08 | 2019-12-17 | 井冈山大学 | rapid breeding method of longya lily bulbs |
-
2020
- 2020-12-18 CN CN202011504519.5A patent/CN112673956A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101061790A (en) * | 2007-04-13 | 2007-10-31 | 浙江省农业科学院 | Quick virus-free group-cultivating and propagating method of OT type lily of hybrid group |
| CN109315288A (en) * | 2017-07-31 | 2019-02-12 | 赵义平 | A kind of tissue culture and rapid propagation method of lily |
| 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 |
| CN110574684A (en) * | 2019-10-08 | 2019-12-17 | 井冈山大学 | rapid breeding method of longya lily bulbs |
Non-Patent Citations (2)
| Title |
|---|
| 苏琼: "龙牙百合快速繁殖研究", 《农技服务》 * |
| 薛建平主编: "《药用植物生物技术》", 31 March 2005, 中国科学技术大学出版社 * |
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