CN113475400B

CN113475400B - Artificial culture technology of cardamine bicolor

Info

Publication number: CN113475400B
Application number: CN202110886520.7A
Authority: CN
Inventors: 胡如芳; 杨亮; 彭招兰; 吴雪松; 周青; 杨伟刚; 周小卿
Original assignee: JI'AN INSTITUTE OF FORESTRY SCIENCE
Current assignee: JI'AN INSTITUTE OF FORESTRY SCIENCE
Priority date: 2021-08-03
Filing date: 2021-08-03
Publication date: 2022-07-12
Anticipated expiration: 2041-08-03
Also published as: CN113475400A

Abstract

The application discloses an artificial cultivation technology of crambe bicolor, which comprises a tissue culture method and an artificial cultivation technology, wherein the artificial cultivation technology comprises the following steps: selecting healthy and strong seedlings with the height of 8-10 cm and at least five fibrous roots at the roots; the plant field needs to ensure the humidity of soil, water in the soil needs to permeate the roots of seedlings, the water temperature is kept between 15 and 20 ℃, and the water in the soil needs to keep fluidity; completely burying the roots of the seedlings in soil, keeping the environmental temperature at 28-32 ℃ in summer and-3-5 ℃ in winter, and selecting the initial planting time at the bottom of two months or at the beginning of three months; the seedlings are in the vegetative stage in 4-6 months, the nutrients are absorbed to grow into young seedlings, the radial leaves on the upper parts of the young seedlings are cut off, the roots of the young seedlings expand, the roots and the stems grow rapidly, and a sunshade is required to be built in summer to ensure that the cardamine hirsute can receive sufficient photosynthesis. The invention aims to realize the purpose of planting the cardamine hirsute by adopting tissue culture and artificial cultivation technologies.

Description

Artificial culture technology of cardamine bicolor

Technical Field

The invention relates to the technical field of plant cultivation, in particular to an artificial cultivation technology of Capsella bursa-pastoris.

Background

Capsella bursa-pastoris belongs to Capsella bursa-pastoris of Cruciferae, and is a perennial herb belonging to wild fibrous plant and growing in stream or shrub forest with elevation of above 1000 m. The root of the cardamine hupingshanesis can be dug and eaten in raw or made into pickled food every 11 months, belongs to a food which is homologous to medicinal food, has strong spicy taste, has the effects of bacteriostasis, disinsection, antisepsis, anticancer and the like, has a relaxed feeling when people suffering from cold and nasal obstruction eat pickled sweat head vegetables and go out of body and sweat, is an inexplicable green healthy food, and has high edible and medicinal values.

The existing cardamine dichotoma grows naturally and under a stream with a higher altitude, and no successful case of artificial planting or artificial cultivation is found, and the technical staff in the field needs to solve the problem of how to artificially cultivate the cardamine dichotoma according to the existing technical means because the cardamine dichotoma has excellent edible and medicinal values.

Disclosure of Invention

In order to solve the problems in the prior art, the invention aims to provide an artificial culture technology for cardamine hirsute, and aims to realize the purpose of planting cardamine hirsute by adopting tissue culture and artificial culture technologies.

In order to realize the purpose of the invention, the technical scheme adopted by the invention is as follows:

an artificial culture technology of cardamine hirsute comprises a tissue culture method and an artificial culture technology, and comprises the following specific steps:

the artificial cultivation technology comprises the following steps:

s1, selecting seedlings: selecting healthy and strong seedlings with the height of 8-10 cm and at least five fibrous roots at the roots;

s2, selecting a site: the plant field needs to ensure the humidity of soil, water in the soil needs to permeate the roots of seedlings, the water temperature is kept between 15 and 20 ℃, and the water in the soil needs to keep fluidity;

s3, planting and cultivating: transplanting the seedlings selected in the step S1 into the soil selected in the step S2, completely burying the roots of the seedlings into the soil, keeping the environmental temperature at 28-32 ℃ in summer and-3-5 ℃ in winter, and selecting the initial planting time at the bottom of two months or at the beginning of three months;

s4, seedling cultivation: the water quality needs to be kept pure in the cultivation process, the seedlings in the vegetative stage in the 4-6 months absorb nutrition to grow into young seedlings, the radial leaves on the upper parts of the young seedlings are cut off, the roots of the young seedlings expand, the roots and the stems grow rapidly, the young seedlings bloom and bear fruits, a sunshade needs to be built in summer to ensure that the cardamine bicolor can receive sufficient photosynthesis, and the roots grow gradually;

the tissue culture method comprises the following steps:

s1, explant collection: taking a stem section of a mother plant of perennial cardamine hirsute which germinates in the current year as an explant, cutting the explant into small sections with at least one axillary bud, and cleaning and disinfecting for later use;

s2, axillary bud induction: firstly, inoculating the explant obtained in the step S1 into a culture medium of MS +6-BA1.0mg/L + IAA0.3mg/L and MS +6-BA1.5mg/L + IAA0.3mg/L, when axillary buds grow to 0.5-3 cm callus, inoculating the callus into the culture medium of MS +6-BA0.2mg/L + IAA0.1mg/L for culturing for 15-20 days, and forming cluster small buds;

s3, propagation culture: and (3) cutting the cluster-shaped small bud clusters in the step (S2) into 3-5 buds/cluster, transferring the cluster-shaped small bud clusters into an MS +6-BA0.1mg/L + IAA0.1mg/L proliferation culture medium for 30 days, carrying out successive generations in a period of 30 days, generating 5-8 sub-buds once for successive generations, selecting a robust single plant with the length higher than 2.5cm from the bud clusters after 3-5 generations of successive generations, transferring the robust single plant into an MS rooting culture medium, wherein 95% of the sub-buds can complete rooting to form seedlings after 40-45 days, and leaves of the seedlings are green.

Optionally, a water circulation device and a water constant temperature device are provided in step S2 of the artificial cultivation technique to ensure the fluidity and constant temperature of water.

Optionally, the seedlings are watered with 0.1% nutrient solution after being transplanted in step S3 of the artificial cultivation technique.

Optionally, in step S4 of the artificial cultivation technique, the sunshade is built when the temperature reaches above 35 ℃.

Optionally, the flowers in step S4 of the artificial cultivation technique are light purple red, and the fruit diameter is 1cm to 2 cm.

Optionally, during winter, the roots of the cardamine hirsute grow greatly, and the leaves of the upper part of the cardamine hirsute are withered.

Optionally, the root comprises a tuber and a fibrous root, the fibrous root growing on the periphery of the tuber, both the tuber and the fibrous root being edible.

Optionally, when the tuber grows to 1kg-2kg or the diameter is 7cm-8cm, digging out the cardamine bicolor for eating or medicine use.

Optionally, small stem blocks grow on the outer periphery of the tuber, and the small stem blocks are picked down and transplanted to the soil selected in the step S2 of the artificial cultivation technology.

Optionally, the disinfecting step in step S2 of the tissue culture method specifically is: taking out the explant, cleaning with sterile water, and cleaning with sterile water on an oscillator for 30-50 min; soaking with alcohol for 0.3-0.8 min, and washing with sterile water for 2-3 times; then sterilizing with 0.1% -0.2% mercuric chloride for 6min-12min, finally washing with sterile water for 3-5 times, and then sucking water with filter paper.

Optionally, after the axillary buds grow to 0.5cm-3cm callus in step S2 of the tissue culture method, the callus is first dark-cultured in an MS culture medium for 18-25 days, and the base is light green and then cultured in a medium inoculated with MS +6-ba0.2mg/L + iaa0.1mg/L.

Optionally, the rooting height of the bud in step S2 of the tissue culture method is 0.4cm-0.6 cm.

Optionally, in step S3 of the tissue culture method, the seedlings need to be placed in a sunshelter to acclimatize for about 1 day, and then transplanted.

The technical scheme of the invention mainly aims at carrying out artificial culture on the Capsella bursa-pastoris, and comprises a tissue culture method and an artificial culture technology, wherein the artificial culture technology mainly comprises selection of a culture environment and a field, firstly, robust seedlings with the height of 8-10 cm and at least five fibrous roots at the roots are selected, secondly, the requirement on the culture environment is strict, flowing water is required, the water temperature is kept at 15-20 ℃, and the roots of the seedlings are completely immersed in the water, so that the roots of the seedlings can be continuously washed by flowing water after being planted, and the roots are always kept clean. In the cultivation process, the environmental temperature should be controlled at 28-32 ℃ in summer, a sunshade should be built when the temperature is too high, the cardamine bicolor can receive enough photosynthesis, and the environmental temperature should be controlled at-3-5 ℃ in winter to avoid freezing. During the growth of seedling, the roots of the seedling grow and expand rapidly except for the growth of upper part of stem and leaf, and the roots can be dug out for eating or medicinal use after the upper part of flower and leaf withers. Furthermore, the seedlings of the invention are obtained by a tissue culture propagation technology, specifically, the stem segments of perennial mother plants which germinate in the current year are selected and placed in a culture medium for tissue culture, the process needs to be carried out by axillary bud induction, bud mass propagation culture, rooting culture and other modes to obtain the required seedlings, and a large amount of seedling propagation can be carried out by the culture mode for cultivation. The crambe bicolor has great medicinal value and edible value, but because the gassing condition is severe, people are not found to plant the crambe bicolor at present, the invention develops an artificial cultivation technology aiming at the crambe bicolor, realizes the planting on the flat ground or according to the existing condition, ensures that people can use the crambe bicolor at any time, and makes great contribution to the society.

Drawings

FIG. 1 shows the tissue culture induced bud of Capsella bursa-pastoris according to the present invention;

FIG. 2 shows the initial planting of the cardamine hirsute of the present invention;

FIG. 3 is a state diagram of the cardamine hirsute of the present invention after 2 months of planting;

FIG. 4 is a state diagram of the cardamine hirsute planted for 10 months.

Detailed Description

The present invention is further illustrated below with reference to specific examples.

Example 1

As shown in fig. 1 to 4, the present embodiment provides an artificial culture technique for cardamine hirsute, including a tissue culture method and an artificial culture technique, and the specific steps are as follows:

the artificial cultivation technology comprises the following steps:

the tissue culture method comprises the following steps:

s3, proliferation culture: and (3) cutting the cluster-shaped small bud clusters in the step (S2) into 3-5 buds/cluster, transferring the cluster-shaped small bud clusters into an MS +6-BA0.1mg/L + IAA0.1mg/L proliferation culture medium for 30 days, carrying out successive generations in a period of 30 days, generating 5-8 sub-buds once for successive generations, selecting a robust single plant with the length higher than 2.5cm from the bud clusters after 3-5 generations of successive generations, transferring the robust single plant into an MS rooting culture medium, wherein 95% of the sub-buds can complete rooting to form seedlings after 40-45 days, and leaves of the seedlings are green.

In this embodiment, Capsella bursa-pastoris is from Capsella bursa-pastoris of Brassicaceae. One to two years old short and small herbage with a height of 10-25 cm, with a branched base and longitudinal grooves. The flower is light purple red, the short siliques are about 1 mm long, the flowering period is 4-5 months, the fruiting period is 5-6 months, the rootstocks are upright, the rootstocks branch from the base, the rootstocks have longitudinal edge grooves, and sometimes the base is blue-purple. The basal leaves are single leaves, the leaves are nearly circular, the length and the width are 1-1.5 cm, the edges are 3-5 shallow to deep cracks, the cracks have irregular round teeth, and the petioles are 2-3 cm long; the lower cauline leaves are nearly circular. The roots of the cardamine dichotoma can be harvested and eaten in raw or made into pickled food every 11 months, belongs to a food which is homologous to medicinal food, has strong spicy taste, has the effects of bacteriostasis, disinsection, antisepsis, anticancer and the like, has a relaxed feeling when people suffering from cold and nasal obstruction eat pickled sweat head vegetables and produce greasy sweat, is a rare green healthy food, and has high edible and medicinal values. The growing conditions of the cardamine hirsute are very strict, and the cardamine hirsute grows under streams or shrubs with the altitude of more than 1000 meters, so that the inventor of the product develops a cultivation technology and can plant the cardamine hirsute in the existing environment.

Specifically, healthy and strong seedlings with the height of 8-10 cm and at least five fibrous roots at the roots are selected during cultivation, and the survival rate of the seedlings can be improved by selecting good seedlings. The roots of the seedlings need to be kept clean in the cultivation process, so that the roots of the seedlings need to be completely immersed in water during cultivation, and the water is guaranteed to have certain activity, namely flowing water is needed, and the water temperature needs to be controlled at 15-20 ℃, so that the survival rate of the cultivation of the cardamine hirsute can be increased. It can be understood that, in order to ensure that the water temperature is controlled at a certain temperature and the water has fluidity, the water can be forced to flow by the water circulation device during cultivation, and meanwhile, the constant temperature and the heating device are arranged to control the water temperature, and certainly, the water purity also needs to be ensured, and the water quality can be regularly detected and replaced. Furthermore, the planting can be performed at a stream mouth or a spring mouth, as long as the conditions required by planting the crambe bicolor can be met.

The technical scheme of the invention mainly aims at cultivating and cultivating the cardamine hirsute, which comprises a tissue culture method and an artificial cultivation technology, wherein the artificial cultivation technology mainly comprises selection of a cultivation environment and a field, firstly, robust seedlings with the height of 8-10 cm and at least five fibrous roots at the roots are selected, secondly, the requirement on the cultivation environment is severe, flowing water is required, the water temperature is kept at 15-20 ℃, and the roots of the seedlings are completely immersed in the water, so that the roots of the seedlings can be continuously washed by flowing water after being planted, and the roots are always kept clean. In the cultivation process, the environmental temperature should be controlled at 28-32 ℃ in summer, a sunshade should be built when the temperature is too high, the cardamine bicolor can receive enough photosynthesis, and the environmental temperature should be controlled at-3-5 ℃ in winter to avoid freezing. During the growth of seedling, the roots of the seedling grow and expand rapidly except for the growth of upper part of stem and leaf, and the roots can be dug out for eating or medicinal use after the upper part of flower and leaf withers. Furthermore, the seedlings of the invention are obtained by a tissue culture propagation technology, specifically, the stem segments of perennial mother plants which germinate in the current year are selected and placed in a culture medium for tissue culture, the process needs to be carried out by axillary bud induction, bud mass propagation culture, rooting culture and other modes to obtain the required seedlings, and a large amount of seedling propagation can be carried out by the culture mode for cultivation. The crambe bicolor has great medicinal value and edible value, but because the gassing condition is severe, people are not found to plant the crambe bicolor at present, the invention develops an artificial cultivation technology aiming at the crambe bicolor, realizes the purpose of planting the crambe bicolor on the flat ground or according to the existing condition, ensures that people can use the crambe bicolor at any time, and makes great contribution to the society.

Because the water quality needs to be kept pure in the cultivation process, and the temperature of the water needs to be kept in a certain range so as to increase the survival rate of the crambe bursaphelenchus xylophilus, the water keeps fluidity through the water circulation device, and meanwhile, the water constant temperature device can ensure that the temperature of the water is in a constant range. Since the water is circulated, the water quality needs to be periodically detected and replaced.

The nutrient solution is added to the seedlings which are just cultivated, so that the local survival rate of the seedlings can be improved, the seedlings can grow more quickly, the adding time of the nutrient solution can be determined according to the situation, the nutrient solution can be added only after the first planting, and the nutrient solution does not need to be added when the later growth is stable.

Optionally, in step S4 of the artificial cultivation technique, the sunshade is built when the temperature reaches above 35 ℃. In the step S4 of the artificial cultivation technology, flowers show light purple red after the plantlets bloom, and the fruit diameter is 1cm-2 cm. When the fruit is in winter, the roots of the cardamine hirsute grow greatly, and the leaves and leaves of the upper part of the cardamine hirsute are in a withered state.

When the temperature is too high, the sun directly irradiates, the photosynthesis of the cardamine hirsute is weakened, and leaves are easy to turn yellow, so that when the temperature reaches above 35 ℃, a sunshade needs to be built to avoid the weakening of the photosynthesis of plants. After the seedlings gradually grow into small seedlings, the seedlings can blossom and bear fruits, the flower colors are light purple, the flower colors are cross-shaped petals, the diameters of the fruits are 1cm-2cm, at the moment, the roots continue to absorb nutrition, the seedlings gradually grow and expand, and the petals and the fruits gradually wither and fall.

In this example, the root includes a tuber and a fibrous root, the fibrous root is grown on the periphery of the tuber, and both the tuber and the fibrous root are edible. When the tuber grows to 1kg-2kg or the diameter is 7cm-8cm, digging out the cardamine bicolor for eating or medicine application.

Particularly, the roots of the cardamine hirsute have great application, tubers and fibrous roots of the cardamine hirsute can be cut into pieces for eating or medicinal use, the eating mode can be pickling or stir-frying for eating, the medicinal mode can be used for soaking wine, the wine can not be whitened, the antibacterial effect is achieved, meanwhile, the cardamine hirsute can be crushed and decocted for medicine, and the effect of treating bronchitis can be achieved both for medicinal use and eating. It should be noted that when the tubers grow to a certain size, the tubers are dug out, so that the edible and medicinal effects of the cardamine bicolor can be enhanced, and particularly, when the tubers grow to 1kg-2kg or the diameter is 7cm-8cm, the digging-out effect is better. The long flower and fruit synchronization segment is needed from planting to maturing of the cardamine hirsute, and the test results are shown in table 1:

TABLE 1 growth of Capsella bursa-pastoris at different stages

As shown in fig. 1, in this embodiment, the sterilization step in step S2 of the tissue culture method specifically includes: taking out the explant, cleaning with sterile water, and cleaning with sterile water on an oscillator for 30-50 min; soaking with alcohol for 0.3-0.8 min, and washing with sterile water for 2-3 times; then sterilizing with 0.1% -0.2% mercuric chloride for 6min-12min, finally washing with sterile water for 3-5 times, and then sucking water with filter paper. After the axillary buds grow to 0.5cm-3cm callus in the step S2 of the tissue culture method, the callus needs to be dark cultured in an MS culture medium for 18 days-25 days, and the base part is light green and then cultured in the MS +6-BA0.2mg/L + IAA0.1mg/L culture medium. The rooting height of the bud in the step S2 of the tissue culture method is 0.4cm-0.6 cm. In step S3 of the tissue culture method, the seedlings need to be put into a sunshelter to be hardened for about 1 day, so that the seedlings adapt to the environmental temperature and then are transplanted.

The stem segments are adopted as the induction objects, so that the method is easy to obtain, short in period, high in rooting rate and easy to survive. Taking young branches of perennial excellent Shuangpai chufa mother plants which germinate in the current year, and cutting the young branches into small sections with the length of about 1cm and axillary buds. The rusty scalpel is not used, the time of exposing the wound in the air is reduced, otherwise, the cut of the stem is browned due to the oxidation of polyphenol oxidase and the like, and the seed production difficulty is increased. The harvested stem segments are put into a refrigerator for about 12 hours after being packaged by a closed container, and then are cleaned by sterile water, and then are cleaned by oscillating on an oscillator for more than 30min by sterile water. Soaking with 70% ethanol for about 0.5min, and washing with sterile water for 2-3 times. Sterilizing with 0.1% mercuric chloride for 6-10min and 0.2% mercuric chloride for 8-12min, washing with sterile water for 3-5 times, sucking dry water with filter paper, inoculating to MS minimal medium, and allowing the sterilized axillary bud to grow to 0.5-3 cm after 40 days. Axillary buds are picked and inoculated into a culture medium of MS +6-BA2.0mg/L + NAA0.2mg/L, after dark culture is carried out for about 20 days, the base parts of the axillary buds form light yellow and green callus with moderate density and good quality, the callus is inoculated into the culture medium of MS +6-BA0.3mg/L + NAA0.1mg/L, and cluster buds appear after culture is carried out for about 15 days. After 30 days, the bud clusters are cut into 3-5 buds/cluster for subculture. Selecting a robust individual plant with the length higher than 2.5cm from a bud mass, dividing a stem section of the robust individual plant into sections with the length of 0.3cm, transferring the stem section into a culture medium of MS +6-BA0.8mg/L + NAA0.2mg/L, carrying out conglobation propagation growth, and carrying out propagation in a cycle of 50 days, wherein each stem section of the robust individual plant can be divided into 4-7 sections, and the propagation multiple can be increased to 9.0-12.0 times. Finally, picking up the strong single buds after propagation, inoculating the strong single buds into a rooting culture medium of 1/2MS + IAA0.3mg/L, wherein the roots begin to sprout after 15-20 days, and after 40-45 days, 95% of seedlings grow 2-3 roots with the length of 0.4-0.6cm, thus completing the propagation of the seedlings.

The present invention is not limited to the above-described alternative embodiments, and various other forms of products can be obtained by anyone in light of the present invention. The above detailed description should not be taken as limiting the scope of the invention, which is defined in the claims, and which the description is intended to be interpreted accordingly.

Claims

1. The culture technology of the cardamine hirsute is characterized by comprising a tissue culture method and an artificial culture technology, and comprises the following specific steps:

the artificial cultivation technology comprises the following steps:

s2, selecting a site: the planting field needs to ensure the humidity of soil, water in the soil needs to permeate the roots of seedlings, the water temperature is kept between 15 ℃ and 20 ℃, and the water in the soil needs to keep fluidity;

s4, seedling cultivation: in the cultivation process, the water quality needs to be kept pure, the seedlings in the vegetative stage in 4-6 months absorb nutrients and grow into young seedlings, the radial leaves on the upper parts of the young seedlings are cut off, at the moment, the roots of the young seedlings are expanded, the roots and stems grow rapidly, the young seedlings bloom and bear fruits, a sunshade needs to be built in summer to ensure that the cardamine marina in two brands can receive enough photosynthesis, and the roots grow gradually;

the tissue culture method comprises the following steps:

s2, axillary bud induction: the explants obtained in step S1 are first inoculated

MS +6-BA1.0mg/L + IAA0.3mg/L and MS +6-BA1.5mg/L + IAA0.3mg/L in culture medium, when axillary bud grows to 0.5-3 cm callus, inoculating the callus to the culture medium

MS +6-BA0.2mg/L + IAA0.1mg/L culture medium for 15-20 days to obtain cluster bud;

2. The technique for cultivating crambe bicolor according to claim 1, wherein a water circulation device and a water constant temperature device are provided in step S2 of the artificial cultivation technique to ensure the fluidity and constant temperature of water.

3. The technique for cultivating crambe bursa-pastoris according to claim 1, wherein the seedling transplantation in step S3 of the artificial cultivation technique requires irrigation with a 0.1% nutrient solution.

4. The technique for cultivating crambe bursa-pastoris according to claim 1, wherein the sunshade is constructed in step S4 when the temperature is above 35 ℃.

5. The cultivation technology of the crambe dichotoma according to claim 1, wherein flowers show light purplish red color after the seedlings bloom in step S4 of the artificial cultivation technology, and the fruit diameter is 1cm-2 cm.

6. The technique for cultivating crambe bicolor according to claim 1, wherein the root comprises a tuber and a fibrous root, the fibrous root grows on the periphery of the tuber, and both the tuber and the fibrous root can be eaten.

7. The technique for culturing cardamine bicolor according to claim 6, wherein when the tuber grows to 1kg-2kg or the diameter is 7cm-8cm, the cardamine bicolor is dug out for eating or medicine application.

8. The technique for cultivating crambe bicolor according to claim 7, wherein small stem blocks grow on the outer periphery of the tubers, and the small stem blocks are picked up and transplanted into the soil selected in step S2 of the artificial cultivation technique.

9. The technique for culturing crambe bicolor according to claim 1, wherein the step of disinfecting in step S1 of the tissue culture method specifically comprises the following steps: taking out the explant, cleaning with sterile water, and cleaning with sterile water on an oscillator for 30-50 min; soaking with alcohol for 0.3-0.8 min, and washing with sterile water for 2-3 times; then sterilizing with 0.1% -0.2% mercuric chloride for 6min-12min, finally washing with sterile water for 3-5 times, and then sucking water with filter paper.

10. The technique for culturing Capsella bursa-pastoris according to claim 1, wherein after axillary buds grow to 0.5cm to 3cm callus in step S2 of the tissue culture method, the callus is dark-cultured in MS medium for 18 days to 25 days to make the base part light green, and then inoculated into MS +6-BA0.2mg/L + IAA0.1mg/L medium for culture.

11. The technique for culturing the crambe dichotoma according to claim 1, wherein the rooting height of the bud in step S3 of the tissue culture method is 0.4cm-0.6 cm.

12. The technique for culturing cardamine hupingshanensis according to claim 1, wherein the seedlings are placed in an awning and hardened for 1 day in step S3 of the tissue culture method, so that the seedlings are suitable for the environmental temperature and then transplanted.