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CN-122004127-A - Tissue culture rapid propagation method for blue rose arrowroot

CN122004127ACN 122004127 ACN122004127 ACN 122004127ACN-122004127-A

Abstract

The invention discloses a tissue culture rapid propagation method of a blue rose arrowroot, which comprises the following steps of 1) obtaining and sterilizing an explant, 2) inducing adventitious buds, 3) propagating and rooting culture, and 4) transplanting test tube seedlings. According to the invention, the new buds of the arrowroot with the blue roses are selected as the explants, the disinfection step is optimized, the modified MS culture medium is selected to replace the common MS culture medium, and the adventitious buds of the explants are induced, proliferated and rooting cultured, so that the disinfection survival rate of the arrowroot, the induction rate of the adventitious buds, the proliferation rate of cluster buds, the rooting rate of seedlings and the transplanting survival rate are remarkably improved, and a large number of test tube seedlings can be propagated in a short time to meet the market demands. The invention has simple and practical technology, feasibility and high application value. The invention can be implemented by simple plant tissue culture equipment.

Inventors

  • ZHANG WEN
  • DU XUELIN
  • Guo Beiyi
  • ZHANG JIFANG
  • ZHONG LIMEI
  • Ruan Sang
  • DAI SEPING
  • YU JIEMEI
  • WU JIANFENG
  • DAI NAN
  • WANG QIANWEN
  • ZHOU HUAN
  • LIU GUOFENG

Assignees

  • 广州市林业和园林科学研究院

Dates

Publication Date
20260512
Application Date
20251229

Claims (8)

  1. 1. The tissue culture and rapid propagation method of the arrowroot with the rose is characterized by comprising the following steps of: a. Selecting a blue rose arrowroot female parent without plant diseases and insect pests, taking newly grown trumpet-shaped tender buds as the explants, digging the tender buds from the underground, not washing the tender buds, cutting off upper leaves, stripping off external bracts, disinfecting 2-3cm stem bases, removing redundant tissues, and inoculating the treated stem bases into an induction culture medium for culture until adventitious buds of the explants grow out, wherein the induction culture medium comprises an improved MS culture medium +3.0-5.0 mg/L6-BA +0.1-0.2mg/L NAA +20-30g/L sucrose +5.8-7g/L agar and pH 5.7-7; b. Cutting off leaves and stems of the obtained adventitious buds, inoculating a bud-retaining base part on a proliferation rooting culture medium for culturing to form cluster buds with roots, wherein the proliferation rooting culture medium comprises an improved MS culture medium, added flower bud No. 1, 2g/L, coconut juice 100 ml/L+2.0-4.0mg/L6-BA+0.1-0.3 mg/L KT+0.1-0.3mg/L NAA+20-30g/L sucrose+6-7 g/L agar and pH 5.8-6.0; c. transplanting test tube seedling, namely transferring the test tube seedling to natural light for hardening off for 3-10 days when the rooting test tube seedling grows to 4-5cm, opening a bottle stopper, taking the test tube seedling out of a culture bottle by using forceps, washing off a root culture medium, and culturing in a matrix; In the step a, the modified MS culture medium is prepared by adjusting the ammonium nitrate concentration to 1900mg/L, the potassium nitrate concentration to 1500mg/L, the magnesium sulfate to 450mg/L and the rest components to be unchanged in a common MS culture medium formula; In the step b, the modified MS culture medium is prepared by adjusting the ammonium nitrate concentration to 1900mg/L, the potassium nitrate concentration to 1500mg/L, the magnesium sulfate to 450mg/L and the rest components to be unchanged in a common MS culture medium formula.
  2. 2. The method according to claim 1, wherein the explant in the step a is obtained by selecting adult blue rose arrowroot sprouts as a material, irrigating once and spraying the leaves with a 500-800 times diluent of 50% carbendazim wettable powder before selecting the explant for sterilization for 42 days, irrigating once and spraying the leaves with 3500-4500 times diluent of agricultural meflounder soluble liquid after 7 days, repeating the above steps for a total of 4 times and 7 days after irrigating the meflounder aqueous solution for the last 1 time, and selecting the treated blue rose arrowroot sprouts as the explant.
  3. 3. The method according to claim 2, wherein the sterilization is performed by wiping the surface of the explant with cotton balls soaked in 75% ethanol by volume fraction, soaking in 75% ethanol for 20-60s after pretreatment, and then continuously sterilizing with 0.1% mercuric solution for several times, and washing with sterile water for 2-6 times after each sterilization to obtain the sterilized explant.
  4. 4. A method according to claim 3, characterized in that the continuous disinfection with 0.1% by mass of the mercuric solution is carried out several times, in particular 3 times with 0.1% by mass of the mercuric solution, the first disinfection time being 4-6 minutes, the second disinfection time being 3-5 minutes, the third disinfection time being 1-3 minutes.
  5. 5. The method according to claim 1, wherein the culture conditions in the step a are that the culture temperature is 22-27 ℃, the illuminance is 500-1000lx and the illuminance is 9-13 h/d.
  6. 6. The method according to claim 1, wherein the cultivation conditions in the step b are cultivation temperature 22-27 ℃, illuminance 1000-2000lx and illuminance 9-13 h/d.
  7. 7. The method of claim 1, wherein the matrix in step c is a mixed matrix of peat soil and perlite in a volume ratio of (2-4): (1-3).
  8. 8. The method of claim 7 wherein the matrix of step c is peat soil and perlite in a volume ratio of 3-4:1-2.

Description

Tissue culture rapid propagation method for blue rose arrowroot Technical Field The invention belongs to the technical field of plant biology, and particularly relates to a tissue culture and rapid propagation method of blue rose arrowroot. Background The genus arrowroot (MARANTACEAE) is a popular foliage plant in the world today, also known as "oil painting plant", belonging to the genus arrowroot (Calathea spp.) of the family arrowroot family. The arrowroot family perennial herb has a bright leaf color due to various varieties and wide application prospect. Traditional methods for arrowroot plant propagation generally comprise plant division propagation and cutting, but have low propagation coefficient and very limited number of propagation seedlings. Therefore, the tissue culture rapid propagation technology becomes an important method for propagating the blue rose arrowroot. The invention adopts the root of the tender bud of the blue rose arrowroot as the explant for tissue culture, and can reproduce a large number of test tube seedlings in a short time so as to meet the market demand. Disclosure of Invention The invention aims at overcoming the defects and provides a tissue culture and rapid propagation method for the arrowroot with blue rose. The invention has simple and practical technology, feasibility and high application value, and can be implemented by using simple plant tissue culture equipment. The technical scheme adopted by the invention is as follows: the tissue culture and rapid propagation method of the arrowroot with the rose is characterized by comprising the following steps of: a. Selecting a blue rose arrowroot female parent without plant diseases and insect pests, taking newly-grown trumpet-shaped tender buds as the explants, digging the tender buds from the underground, not washing the tender buds, cutting off upper leaves, stripping off external bracts, disinfecting 2-3cm stem bases, removing redundant tissues, inoculating the treated stem bases into a culture medium for culturing until adventitious buds of the explants grow out, wherein the induction culture medium comprises an improved MS culture medium +3.0-5.0 mg/L6-BA +0.1-0.2mg/L NAA +20-30g/L sucrose +5.8-7/L agar and pH 5.7-7; a. Cutting off leaves and stems of the obtained adventitious buds, inoculating a bud-retaining base part on a proliferation rooting culture medium for culturing to form cluster buds with roots, wherein the proliferation rooting culture medium comprises an improved MS culture medium, added flower bud No. 1, 2g/L, coconut juice 100 ml/L+2.0-4.0mg/L6-BA+0.1-0.3 mg/L KT+0.1-0.3mg/L NAA+20-30g/L sucrose+6-7 g/L agar and pH 5.8-6.0; b. Transplanting test tube seedling, namely transferring the test tube seedling to natural light for hardening off for 3-10 days when the rooting test tube seedling grows to 4-5cm, opening a bottle stopper, taking the test tube seedling out of a culture bottle by using forceps, washing off a root culture medium, and culturing in a matrix; In the step a, the modified MS culture medium is prepared by adjusting the ammonium nitrate concentration to 1900mg/L, the potassium nitrate concentration to 1500mg/L, the magnesium sulfate to 450mg/L and the rest components to be unchanged in a common MS culture medium formula; In the step b, the modified MS culture medium is prepared by adjusting the ammonium nitrate concentration to 1900mg/L, the potassium nitrate concentration to 1500mg/L, the magnesium sulfate to 450mg/L and the rest components to be unchanged in a common MS culture medium formula. Preferably, the explant in the step a is obtained by a pretreatment method comprising the steps of selecting adult blue rose arrowroot sprouts as a material, irrigating once with 500-800 times of 50% carbendazim wettable powder and spraying the leaves before 42 days before the explant is selected for sterilization, irrigating once with 3500-4500 times of agricultural mevalonate soluble liquid and spraying the leaves after 7 days, repeating the steps for 4 times after 7 days, irrigating 7 days after the final time of irrigating with a mevalonate aqueous solution, and selecting the treated blue rose arrowroot sprouts as the explant. Preferably, the sterilization is to wipe and sterilize the surface of the explant by cotton balls soaked by 75% ethanol by volume fraction, soak the surface in 75% ethanol for 20-60s after pretreatment, and then sterilize the surface of the explant by 0.1% mercuric chloride solution for several times continuously, and rinse the surface of the explant by sterile water for 2-6 times after each sterilization. Preferably, the continuous disinfection for a plurality of times by using the 0.1 percent mercuric solution by mass fraction is specifically that the continuous disinfection for 3 times by using the 0.1 percent mercuric solution by mass fraction is carried out, wherein the first disinfection time is 4-6 minutes, the second disinfection time is 3-5 minutes, and the thi