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A kind of induction method of Thyme thyme callus

A technology of callus induction and callus, applied in horticultural methods, botanical equipment and methods, plant regeneration, etc., can solve the problems of small reproduction coefficient, less flowering, and inability to meet large-scale production

Active Publication Date: 2021-09-10
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, the propagation method of Trichosanthes is mainly based on seed propagation. In the wild state, Trismus has less flowering, resulting in a low seed setting rate. Therefore, the reproductive coefficient of propagation by seeds is too small to meet the needs of large-scale production.

Method used

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  • A kind of induction method of Thyme thyme callus
  • A kind of induction method of Thyme thyme callus
  • A kind of induction method of Thyme thyme callus

Examples

Experimental program
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Effect test

preparation example Construction

[0036] Preparation of culture medium and culture conditions: The test designed to MS medium and different concentrations of NAA needed, 2,4-D, 6-BA and other plant growth regulators, agar 8g / L, sucrose 30g / L, with 1mol / L NaOH and HCl in the medium was adjusted to pH 5.8 to 6.0, boil, until the agar melted and dispensed into tissue culture flasks, autoclaved at 121 deg.] C 25min, remove the stand.

[0037] The culture conditions are: fluorescent lighting, the culture temperature is 23 ~ 25 ℃, relative humidity 50-70%, illumination time 12h / d, the light intensity of 1500 ~ 2000Lx.

[0038] Statistics and data analysis: Excel 2007 with data processing and mapping, analysis of variance SPSS 20 software for data analysis.

[0039] Contamination rate (%) = Number of seed contamination / inoculation of seed number × 100%

[0040] Number of seed germination (%) = Number germination / seed uncontaminated × 100%

[0041] Browning rate (%) = Number of browning / inoculated explant mass...

Embodiment 1

[0043] Example 1 Effect of sterilization time on different embodiment seed disinfection results

[0044] In the clean table, the Trinidad Hong mature seeds first with 75% ethanol solution disinfection 1min, and then containing 0.1% Tween 20 HgCl 2 Solution is sterilized 6 ~ 12min, Tween 20: HgCl 2 Solution (v / v) = 1: 1000, sterile water rinsed 6 times, each time 3min; culture flasks set into the standby lid sterilized seeds are sterilized, disinfected before each inoculation good seed discharge into a Petri dish covered with sterile filter paper, the material surface water drain, scalpel and tweezers removal of the seed coat, and then inoculated to MS basal medium. Each treatment followed by at least 15 bottles of two connected seed, was repeated three times. After inoculation 20d statistics contamination rate and seed germination rate, compare disinfection process and its effect on seed germination.

[0045] Trinidad Hong fruits and seeds such as pictures figure 1 , The seed ge...

Embodiment 2

[0050] Effect of different concentrations of hormone types Example 2 Trinidad Hong embodiment callus induction

[0051] Inducing callus explant is no vaccine blade root section. Selection cytokinin 6-BA (0.3mg / L, 0.5mg / L, 1.0mg / L), auxin 2,4-D (0.5mg / L, 1.0mg / L, 2.0mg / L) and of NAA ( 1.0mg / L, 2.0mg / L, 3.0mg / L) orthogonal to callus inducing hormone, respectively, with 6-BA of NAA, 6-BA and 2,4-D is a 3 factorial design 2 factors , to give a total of 18 formulations, each treatment at least 15 explants, repeated 3 times for each formulation. Callus was first observed regularly in the days, callus induction, callus state color, select the most suitable Trinidad Hong callus induction material, media formulations.

[0052] Orthogonal experimental design in Table 2. Trinidad Hong Callus

[0053]

[0054] 1, different hormone types and concentrations on callus induction of Trinidad Hong aseptic blade

[0055] After aseptic blades cut into an appropriate size and inocu...

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Abstract

The invention discloses a callus induction method of Thyme thyme. The induction method includes the following steps: S1. Pretreatment of materials: first sterilize the mature seeds of Thyme thyme with 75% ethanol solution for 1-2 minutes, then sterilize them with mercuric chloride containing Tween 20 for 6-12 minutes, and sterilize the sterilized The seed coats of the seeds were removed and inoculated into MS basic medium to cultivate aseptic seedlings; S2. Callus induction: inoculate the leaves or roots of aseptic seedlings as explants on the induction medium and cultivate them until the callus grows. damage tissue. The present invention uses the leaves or root sections of aseptic seedlings as explants to study the induction medium suitable for leaf or root section callus respectively, which can reduce the browning rate while improving the callus induction rate of Thyme thyme , and does not affect the subsequent proliferation and subculture of callus, it can be applied to the breeding of Thyme chinensis seedlings, induction of polyploidy, induction of mutations and innovation of transgenic germplasm.

Description

Technical field [0001] The present invention relates to the field of tissue culture techniques, in particular, it relates to a method of inducing callus Trinidad Hong. Background technique [0002] Trinidad Hong (Murraya paniculata (L.) Jack.) Rutaceae (Rutaceae) murraya (Murraya) plant, also known as gold-gui, holly, green wood, etc., produced in Taiwan, Fujian, Guangdong, Guangxi and other places. Trinidad Hong is one of the main raw material known Chinese medicine "Sanjiuweitai," the 2015 edition of "Chinese Pharmacopoeia" collection contains herbal medicines Murraya paniculata plant sources M.exotica L. and Trinidad Hong M.paniculata (L. ) Jack., both the leaves and twigs with leaves as paniculata medicine, with swelling and pain, efficacy of blood stasis. In fact Sanjiuweitai particles and capsules paniculata herbs used are derived from the leaves and twigs Trinidad Hong leafy. [0003] At present the majority of studies on Trinidad Hong focused on chemical constituents and ...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 何国振蒋烨
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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