CN111084108B - Culture method for tissue culture of floral leaf dwarf pampasgrass - Google Patents
Culture method for tissue culture of floral leaf dwarf pampasgrass Download PDFInfo
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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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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/10—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material
- A01G24/12—Growth substrates; Culture media; Apparatus or methods therefor based on or containing inorganic material containing soil minerals
- A01G24/15—Calcined rock, e.g. perlite, vermiculite or clay aggregates
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
- A01G24/20—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material
- A01G24/28—Growth substrates; Culture media; Apparatus or methods therefor based on or containing natural organic material containing peat, moss or sphagnum
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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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Abstract
The invention belongs to the technical field of plant tissue culture, and particularly relates to a culture method for tissue culture of floral leaf dwarf pampas grass. The culture method for tissue culture of the floral leaf dwarf pampasgrass comprises the following steps: selection of explants, disinfection and sterilization of explants, bud induction culture, subculture, rooting culture and transplantation and seedling hardening. The method combines the growth characteristics of the floral leaf dwarf pampasgrass, adopts high and low concentration hormones to alternately proliferate by selecting a proper disinfection mode, culture medium and seedling hardening mode, reduces vitrification and effectively improves proliferation coefficient, can solve the problem of serious vitrification caused by high concentration hormones during proliferation culture, has simple, novel and efficient seedling hardening mode, can greatly reduce production cost, can efficiently and economically obtain a large amount of excellent tissue culture seedlings of the floral leaf dwarf pampasgrass in a certain time, and has wide market application prospect.
Description
Technical Field
The invention belongs to the technical field of plant tissue culture, and particularly discloses a culture method for tissue culture of floral leaf dwarf pampasgrass.
Technical Field
The floral leaf dwarf pampasgrass is a perennial herbaceous plant with the plant height of 120cm, leaves gathered at the base part, is long and narrow, has fine teeth on the edge, has large conical inflorescence, is feather-shaped, is silvery white, is a foreign famous ornamental plant, and is used for landscaping or bank planting. Positive, enjoys sunny, warm and humid environment, and is also cold-resistant; the method has the advantages of strong adaptability, no soil selection, water-moisture resistance and drought resistance, large demand on the market, and many reports on tissue culture research of the floral leaf dwarf pampasgrass in China at present, but the defects of unstable proliferation proportion, unstable heredity, difficult leaf length operation during propagation and transfer, difficult seedling hardening and cleaning, difficult planting, high production cost and the like exist.
Disclosure of Invention
The invention aims to solve the technical problem of providing a culture method for tissue culture of floral leaf dwarf pampasgrass aiming at the defects of the prior art. The invention adopts high and low concentration hormones to alternately proliferate and propagate with high proliferation ratio, can improve the propagation speed and the uniformity of seedlings and improve the stability of characters, thereby establishing an industrial production system more suitable for the rapid propagation of floral leaf dwarf pamphlet.
In order to solve the technical problems, the invention adopts the technical scheme that: a culture method for tissue culture of pampas grass is characterized by comprising the following steps:
(1) selection and sterilization of explants: selecting a strong and disease-free mosaic and dwarf pampasgrass plant, selecting a newly grown tender bud as an explant, removing redundant stem segments and wrapped leaves of the explant, washing off attached broken leaves and impurities, putting the treated explant into a mercuric chloride solution which is mixed with 1-2 drops of Tween and has the mass fraction of 0.08% -0.12%, sterilizing for 7-9 minutes, taking out, washing with sterile water for 4-6 times, cutting off brown stains, cutting the explant into 2-3 cm small segments, inoculating the small segments to a primary culture medium, and culturing for 5-7 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-pollution explant from a primary culture medium, inoculating the explant to an induction culture medium, forming an adventitious bud in 7-15 days, and increasing the height of the adventitious bud to 2-3 cm after culturing for 15-25 days, wherein the induction culture medium is an MS culture medium containing 0.5-1.5 mg/L of 6-BA, 0.4-0.6 mg/L of 2, 4-D, sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) to a multiplication culture medium, wherein the multiplication culture medium is an MS culture medium containing high-concentration hormone 6-BA or low-concentration hormone 6-BA, sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuing culturing, wherein the culture medium is 1/4MS culture medium containing 0-1.0 mg/L IBA, sucrose and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25-30 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 2-3 minutes, cleaning and planting in a peat and perlite mixed matrix, putting the seedlings into a greenhouse after planting, keeping the temperature at 15-25 ℃ and the humidity at over 65%.
In the step (3), 2.0-3.0 mg/L of high-concentration hormone 6-BA and 0.2-0.4 mg/L of low-concentration hormone 6-BA are alternately used in the whole multiplication culture process, and the multiplication coefficient of propagation is controlled to be more than 8.
In the step (4), the seedlings grow root primordium in 7-15 days during rooting culture, the roots grow 3-4 cm in 15-25 days, and the rooting rate is 95% -100%.
In the steps (2) - (4), the axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium contain 20-30 g/L of sucrose and 5-7 g/L of agar powder, and the pH value of the culture medium is 5.8-6.2.
In the steps (2) - (4), in the processes of bud induction, proliferation and rooting culture, the culture environment temperature is 24-26 ℃, and the illumination is 1500-2500 lx.
The proportion of the mixed matrix peat and the perlite in the seedling hardening in the step (5) is 3: 0.5-2.
And (5) watering for 10-15 seconds every half hour from 8 o 'clock to 6 o' clock at night after the transplanting is finished, keeping the soil of the nursery stock moist, continuing for 6-8 days, removing the sunshade net after one week, watering once a day for 8-12 minutes, and ventilating once.
Compared with the prior art, the invention has the following advantages:
the culture method of the tissue culture of the floral leaf dwarf pampasgrass combines the growth characteristics of the floral leaf dwarf pampasgrass, selects a proper disinfection mode, culture medium and seedling exercising mode, adopts high-concentration and low-concentration hormones to alternately proliferate, reduces vitrification, effectively improves proliferation coefficient, has simple, novel and efficient seedling exercising mode and can greatly reduce production cost in order to solve the problems of low proliferation proportion, long seedling leaves, difficult proliferation and transfer, difficult rooting and seedling exercising and the like caused by accumulation of high-concentration hormones and low-concentration hormones during proliferation culture.
Example 1
A culture method for tissue culture of pampas grass comprises the following steps:
(1) selection and sterilization of explants: selecting strong and disease-free floral leaf dwarf pampasgrass plants, selecting fresh buds as explants, removing redundant stem segments and wrapped leaves of the explants, washing the explants in clear water, washing off broken leaves and impurities attached to the broken leaves, putting the treated explants into 0.08 percent mercuric chloride solution mixed with 1 drop of Tween, sterilizing for 7 minutes, taking out the explants, washing the explants with sterile water for 4 times, cutting off brown stain parts, cutting the explants into 2cm small segments, inoculating the small segments to a primary culture medium, and culturing for 5 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) bud induction culture: selecting a non-polluted explant from a primary culture medium, inoculating the explant to an induction culture medium, forming an adventitious bud after 15 days, and increasing the height of the adventitious bud to 2cm after 15 days of culture, wherein the induction culture medium is an MS culture medium containing 6-BA (0.5 mg/L), 2, 4-D (0.4 mg/L), sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) onto a multiplication culture medium, wherein high-concentration hormones and low-concentration hormones are alternately used in the whole multiplication culture process, so that the multiplication coefficient is more than 8, and the multiplication culture medium is an MS culture medium containing high-concentration hormone 6-BA (2.0 mg/L), low-concentration hormone 6-BA (0.2 mg/L), sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuing culturing, wherein the seedlings grow root primordium in 15 days, the roots grow 3-4 cm in 30 days, the rooting rate is 95%, and the rooting culture medium is 1/4MS culture medium containing cane sugar and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 2 minutes, cleaning and planting in a peat and perlite mixed matrix, wherein the ratio of the peat to the perlite mixed matrix is 3:0.5, placing in a greenhouse after planting, watering for 10 seconds every half hour from 8 o 'clock to 6 o' clock after transplanting is completed, keeping the seedling soil moist for 6 days, removing a sunshade net, watering once a day for 8 minutes, ventilating once, keeping the temperature at 15 ℃ and the humidity at more than 65%.
The axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium further comprise 20g/L of sucrose and 5g/L of agar powder, the pH value of the culture medium is 5.8, the culture temperature is 24 ℃, and the illumination is 1500 lx.
Example 2
A culture method for tissue culture of pampas grass comprises the following steps:
(1) selection and sterilization of explants: selecting strong and disease-free floral leaf dwarf pampasgrass plants, selecting fresh buds as explants, removing redundant stem segments and wrapped leaves of the explants, washing the explants in clear water, washing off broken leaves and impurities attached to the leaves, putting the treated explants into 0.09% mercuric chloride solution mixed with 1 drop of Tween, sterilizing for 7 minutes and 30 seconds, taking out the explants, washing the explants with sterile water for 5 times, cutting off brown stains, cutting the explants into 2.5cm small segments, inoculating the small segments to a primary culture medium, and culturing for 9 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-polluted explant from a primary culture medium, inoculating the explant onto an induction culture medium, after 18 days, carrying out bud meristem culture, and then culturing for 18 days, wherein the adventitious bud grows to 2-3 cm, and the induction culture medium is an MS culture medium containing 6-BA (0.75 mg/L), 2, 4-D (0.4 mg/L), sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) onto a multiplication culture medium, wherein high-concentration hormones and low-concentration hormones are alternately used in the whole multiplication culture process, so that the multiplication coefficient is more than 8, and the multiplication culture medium is an MS culture medium containing high-concentration hormone 6-BA (2.25 mg/L), low-concentration hormone 6-BA (0.25 mg/L), sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuously culturing, wherein 9 days of seedlings grow root primordium, 18 days of seedlings grow roots with the length of 3-4 cm, and the rooting rate is 96%. The culture medium is 1/4MS culture medium containing IBA (0.25 mg/L), sucrose and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 2 minutes, cleaning and planting in a mixed matrix of peat and perlite, wherein the ratio of the mixed matrix peat to the perlite is 3:1, placing in a greenhouse after planting, watering for 12 seconds every half hour from 8 o 'clock to 6 o' clock after the completion of transplanting, keeping the soil of the seedlings moist for 6 days, removing a sunshade net, watering once a day for 9 minutes, ventilating once, and keeping the temperature at 18 ℃ and the humidity at over 65%.
The axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium further comprise 20g/L of sucrose and 5g/L of agar powder, the pH value of the culture medium is 5.8, the culture temperature is 24 ℃, and the illumination is 1500 lx.
Example 3
A culture method for tissue culture of pampas grass comprises the following steps:
(1) selection and sterilization of explants: selecting strong and disease-free floral leaf dwarf pampasgrass plants, selecting fresh buds as explants, removing redundant stem segments and wrapped leaves of the explants, washing the explants in clear water, washing off broken leaves and impurities attached to the leaves, putting the treated explants into 0.10 percent mercuric chloride solution mixed with 1.5 drops of Tween, sterilizing for 8 minutes, taking out the explants, washing the explants with sterile water for 5 times, cutting off brown stains, cutting the explants into 2.5cm small segments, inoculating the small segments to a primary culture medium, and culturing for 11 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-polluted explant from a primary culture medium, inoculating the explant to an induction culture medium, after 20 days, carrying out bud meristem culture, and then culturing for 20 days until an adventitious bud grows to 2-3 cm, wherein the induction culture medium is an MS culture medium containing 6-BA (1.00 mg/L), 2, 4-D (0.5 mg/L), sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to propagation culture in the step (2) onto a propagation culture medium, wherein high-concentration hormones and low-concentration hormones are alternately used in the whole propagation culture process, so that the propagation coefficient is more than 8, and the propagation culture medium is an MS culture medium containing high-concentration hormone 6-BA (2.50 mg/L), low-concentration hormone 6-BA (0.30 mg/L), sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuously culturing, wherein the seedlings grow root primordium in 11 days, the roots grow 3-4 cm in 20 days, the rooting rate is 97%, and the rooting culture medium is 1/4MS culture medium containing IBA (0.50 mg/L), cane sugar and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 3 minutes, cleaning and planting in a mixed matrix of peat and perlite, wherein the ratio of the mixed matrix peat to the perlite is 3:1, placing in a greenhouse after planting, watering for 13 seconds every half hour from 8 o 'clock to 6 o' clock after the completion of transplanting, keeping the seedling soil moist for 7 days (covering a sunshade net externally), removing the sunshade net, watering once a day for 10 minutes, ventilating once, keeping the temperature at 20 ℃ and the humidity at more than 65%.
The axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium further comprise 25g/L of sucrose and 6g/L of agar powder, the pH value of the culture medium is 6.0, the culture temperature is 25 ℃, and the illumination is 2000 lx.
Example 4
A culture method for tissue culture of pampas grass comprises the following steps:
(1) selection and sterilization of explants: selecting strong and disease-free floral leaf dwarf pampasgrass plants, selecting fresh buds as explants, removing redundant stem segments and wrapped leaves of the explants, washing the explants in clear water, washing off broken leaves and impurities attached to the leaves, putting the treated explants into 0.11% mercuric chloride solution mixed with 2 drops of Tween, sterilizing for 8 minutes and 30 seconds, taking out the explants, washing the explants with sterile water for 6 times, cutting off brown stains, cutting the explants into 3cm small segments, inoculating the small segments to a primary culture medium, and culturing for 13 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-polluted explant from a primary culture medium, inoculating the explant to an induction culture medium, after 23 days, obtaining a meristem of a bud, and after 23 days of culture, growing an adventitious bud to 2-3 cm, wherein the induction culture medium is an MS culture medium containing 6-BA (1.25 mg/L), 2, 4-D (0.5 mg/L), sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) onto a multiplication culture medium, wherein high-concentration hormones and low-concentration hormones are alternately used in the whole multiplication culture process, so that the multiplication coefficient is more than 8, and the multiplication culture medium is an MS culture medium containing high-concentration hormone 6-BA (2.75 mg/L), low-concentration hormone 6-BA (0.35 mg/L), sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuing to culture, wherein the seedlings grow root primordium in 13 days, the roots grow 3-4 cm in 22 days, the rooting rate is 98%, and the rooting culture medium is 1/4MS culture medium containing IBA (0.75 mg/L), cane sugar and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 3 minutes, cleaning and planting in a mixed matrix of peat and perlite, wherein the ratio of the mixed matrix peat to the perlite is 3:1.5, placing in a greenhouse after planting, watering for 14 seconds every half hour from 8 o 'clock to 6 o' clock after the completion of transplanting, keeping the seedling soil moist for 8 days (covering a sunshade net externally), removing the sunshade net, watering once a day for 11 minutes, ventilating once, keeping the temperature at 23 ℃ and the humidity at more than 65%.
The axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium further comprise 30g/L of sucrose and 7g/L of agar powder, the pH value of the culture medium is 6.2, the culture temperature is 26 ℃, and the illumination is 2500 lx.
Example 5
A culture method for tissue culture of pampas grass comprises the following steps:
(1) selection and sterilization of explants: selecting strong and disease-free floral leaf dwarf pampasgrass plants, selecting fresh buds as explants, removing redundant stem segments and wrapped leaves of the explants, washing the explants in clear water, washing off broken leaves and impurities attached to the broken leaves, putting the treated explants into 0.12% mercuric chloride solution mixed with 2 drops of Tween, disinfecting for 9 minutes, taking out and washing with sterile water for 6 times, cutting brown stains, cutting the explants into 3cm small segments, inoculating the small segments to a primary culture medium, and culturing for 15 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-polluted explant from a primary culture medium, inoculating the explant to an induction culture medium, after 25 days, obtaining a meristem of a bud, and after 25 days of culture, growing an adventitious bud to 2-3 cm, wherein the induction culture medium is an MS culture medium containing 6-BA (1.50 mg/L), 2, 4-D (0.6 mg/L), sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) onto a multiplication culture medium, wherein high-concentration hormones and low-concentration hormones are alternately used in the whole multiplication culture process, so that the multiplication coefficient of propagation is more than 8, and the multiplication culture medium is an MS culture medium containing high-concentration hormones 6-BA (3.00 mg/L), low-concentration hormones 6-BA (0.40 mg/L), sucrose and agar;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuously culturing, wherein the seedlings grow root primordium in 15 days, the roots grow 3-4 cm in 25 days, the rooting rate is 100%, and the rooting culture medium is 1/4MS culture medium containing IBA (1.00 mg/L), cane sugar and agar;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 3 minutes, cleaning and planting in a mixed matrix of peat and perlite, wherein the ratio of the mixed matrix peat to the perlite is 3:2, placing in a greenhouse after planting, watering for 15 seconds every half hour from 8 o 'clock to 6 o' clock after the completion of transplanting, keeping the soil of the seedlings moist for 8 days, removing a sunshade net, watering once a day for 12 minutes, ventilating once, and keeping the temperature at 25 ℃ and the humidity at more than 65%.
The axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium further comprise 30g/L of sucrose and 7g/L of agar powder, the pH value of the culture medium is 6.2, the culture temperature is 26 ℃, and the illumination is 2500 lx.
The above description is only for the preferred embodiment of the present invention, and is not intended to limit the present invention in any way. Any simple modification, change and equivalent changes of the above embodiments according to the principles of the present invention are still within the protection scope of the technical solution of the present invention.
Claims (5)
1. A culture method for tissue culture of pampas grass is characterized by comprising the following steps:
(1) selection and sterilization of explants: selecting a strong and disease-free mosaic and dwarf pampasgrass plant, selecting a newly grown tender bud as an explant, removing redundant stem segments and wrapped leaves of the explant, washing off attached broken leaves and impurities, putting the treated explant into a mercuric chloride solution which is mixed with 1-2 drops of Tween and has the mass fraction of 0.08% -0.12%, sterilizing for 7-9 minutes, taking out, washing with sterile water for 4-6 times, cutting off brown stains, cutting the explant into 2-3 cm small segments, inoculating the small segments to a primary culture medium, and culturing for 5-7 days, wherein the primary culture medium is 1/2MS culture medium containing sucrose and agar powder;
(2) and (3) bud induction culture: selecting a non-pollution explant from a primary culture medium, inoculating the explant to an induction culture medium, forming an adventitious bud in 7-15 days, and increasing the height of the adventitious bud to 2-3 cm after culturing for 15-25 days, wherein the induction culture medium is an MS culture medium containing 0.5-1.5 mg/L of 6-BA, 0.4-0.6 mg/L of 2, 4-D, sucrose and agar;
(3) and (3) proliferation culture: inoculating the adventitious buds subjected to bud induction culture in the step (2) onto a multiplication culture medium, wherein the multiplication culture medium is an MS culture medium containing high-concentration hormone 6-BA or low-concentration hormone 6-BA, sucrose and agar, the high-concentration hormone 6-BA is used alternatively at 2.0-3.0 mg/L and the low-concentration hormone 6-BA is used alternatively at 0.2-0.4 mg/L in the whole multiplication culture process, and the multiplication coefficient of the propagation is controlled to be more than 8;
(4) rooting culture: dividing the seedlings after the propagation culture into single plants, inoculating the single plants into a rooting culture medium, and continuously culturing, wherein the culture medium is 1/4MS culture medium containing 0-1.0 mg/L IBA, cane sugar and agar, the seedlings grow root primordium in 7-15 days during the rooting culture, the roots grow 3-4 cm in 15-25 days, and the rooting rate is 95% -100%;
(5) hardening seedlings: taking out seedlings cultured on a rooting culture medium for 25-30 days, removing roots, washing the culture medium, soaking in a carbendazim solution for 2-3 minutes, cleaning and planting in a peat and perlite mixed matrix, putting the seedlings into a greenhouse after planting, keeping the temperature at 15-25 ℃ and the humidity at over 65%.
2. The method for tissue culture of floral leaf dwarf pampasgrass according to claim 1, wherein in the steps (2) - (4), the axillary bud induction culture medium, the proliferation culture medium and the rooting culture medium contain 20-30 g/L of sucrose and 5-7 g/L of agar powder, and the pH of the culture medium is 5.8-6.2.
3. The method for tissue culture of floral leaf dwarf pampasgrass according to claim 1, wherein in the steps (2) - (4), the culture environment temperature is 24-26 ℃ and the illumination is 1500-2500 lx in the bud induction, proliferation and rooting culture processes.
4. The method for tissue culture of pampas grass according to claim 1, wherein the ratio of peat and perlite as the mixed substrate during seedling hardening in step (5) is 3: 0.5-2.
5. The tissue culture method of floral leaf dwarf pampasgrass according to claim 1, wherein during hardening off the seedlings in step (5), watering is performed every half hour for 10-15 seconds from 8 o 'clock to 6 o' clock in the morning after transplanting is completed, keeping the seedling soil moist for 6-8 days, removing the sunshade net after one week, watering once a day for 8-12 minutes, and ventilating once.
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|---|---|---|---|---|
| CN101869053A (en) * | 2009-04-23 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of floral leaf pampasgrass |
| CN101869058A (en) * | 2009-04-23 | 2010-10-27 | 上海上房园林植物研究所 | Tissue culture method of short pampas grass |
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| US20160050866A1 (en) * | 2014-08-20 | 2016-02-25 | The Institute For Advanced Learning And Research | Micropropagation and plant regeneration systems for arundo donax and other monocots |
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