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Method for establishing paeonia lactiflora in-vitro regeneration system

A technology of in vitro regeneration and method establishment, which is applied in the field of peony tissue culture propagation, which can solve the problems of long cycle, inability to fully maintain the traits of the female parent, and limited number of underground dormant buds

Inactive Publication Date: 2018-09-28
蒋建华
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

CN102428872 discloses a method for obtaining complete peony plants through seed embryo culture, but because the seed embryo cannot fully maintain the traits of the female parent, it cannot be used for the propagation of fine varieties
The papers published by Wu Hongjuan et al.: Induction and rooting technology of ornamental peonies 'Da Fugui' cluster buds, 'Comparison of different varieties of peonies initiating culture and 'Zhushapan' cluster bud induction' reported that underground dormant buds were used as explants to induce clusters Bud method, this method has the following deficiencies for the propagation of good varieties: 1. the number of underground dormant buds contained in a plant is limited, and the propagation efficiency with it as explants is not high; 2. every strain of tissue culture seedlings can only produce 3 ~5 clustered buds, the reproduction coefficient is not high; ③The whole process must go through a process of starting and cultivating underground buds, which consumes a lot of manpower and material resources and increases the cost
To sum up, in the process of peony tissue culture, the existing techniques have successfully differentiated adventitious buds, but they mostly use seed embryos, underground dormant buds, stem segments, etc. as explants, not only the differentiation rate is low, but the number of subcultures Many, the experimental process is cumbersome, the cycle is long, and the desired effect cannot be obtained

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  • Method for establishing paeonia lactiflora in-vitro regeneration system

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Experimental program
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Effect test

Embodiment 1

[0032] 1 Example of medium screening

[0033] During the establishment of the in vitro regeneration system of the callus pathway, the success of callus differentiation, that is, the induction medium is a key step in the regeneration system. The rate of callus differentiation directly affects whether the regenerative tissue culture system is feasible. The combination of basic medium and hormone types and concentrations in the callus differentiation medium has a great correlation with the degree and rate of callus differentiation. The present invention has tried different basic medium and different hormone concentration ratios, and the influence on the degree of induction of leaves of Paeoniae Alba is shown in Table 1. It can be seen from Table 1 that MS+sucrose 30g / L+KT 1mg / L+IBA0.02mg / L, the callus differentiation rate of Hangbai Shao is the highest, which is the best callus differentiation medium.

[0034] Table 1

[0035] Types of basal medium

Hormone combina...

Embodiment 2

[0042] (1) Obtaining explants: Select young and tender leaves of Paeoniae Alba from late March to early April, rinse them with clean water, then rinse them once with Amway detergent, shake them on a shaker for 15-20 minutes; rinse them with running water for 3 minutes, Sterilize with 75% ethanol for 10-20s on the sterile operating table, pour 0.1% mercuric chloride immediately, and finally wash twice with sterile water, and cut the sterilized leaves into 2cm long pieces;

[0043] (2) Callus preparation: take the explant segments obtained in step (1), inoculate them in the induction medium for callus induction, insert a total of 40 leaf segments, and culture at 24-28°C , first cultivated in dark for 7-10d, and then cultivated in light for 25d; the light culture conditions are: the light time is 14h / d, and the light intensity is 1500lx; the induction medium is MS+sucrose 30g / L+KT 1mg / L+ IBA0.02mg / L, pH value 5.4-6.0; there are 35 green calluses at both ends of the explants, and ...

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Abstract

The invention discloses a method for establishing a paeonia lactiflora in-vitro regeneration system. The system is characterized by comprising the following steps: obtaining an explant, preparing callus, carrying out differentiation of adventitious buds, carrying out proliferation, carrying out root induction, carrying out seedling hardening, and carrying out transplanting. According the application, the in-vitro regeneration system of a paeonia lactiflora variety of paeoniae alba is systematically established, and the application is one of the important ways to solve the shortage of peony seedlings, has very important significance for the south domestication of the paeonia lactiflora and the development, promotion and utilization of the paeonia lactiflora, and has a wide application prospect.

Description

technical field [0001] The invention relates to a propagation method of peony tissue culture, belongs to the technical field of seedling propagation of herbaceous ornamental plants, and in particular relates to a method for establishing a peony regeneration system in vitro. Background technique [0002] Paeonia lactiflora (scientific name: Paeonia lactiflora Pall.), aliased as parting grass and prime minister of flowers, belongs to the order of syringa, and is a perennial herb of the Paeoniaceae Paeoniaceae group. The tuberous root is produced from the bottom of the root neck, fleshy, strong, spindle-shaped or long columnar, 0.6-3.5cm thick. The petals of peony are obovate, and the disk is shallow cup-shaped. The flowering period is from May to June. The flowers are usually born on the top of the stem or near the top of the leaf axil. The original species of flowers are white, with 5 to 13 petals. The horticultural varieties are rich in flower colors, including white, pink,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 蒋建华周小成许奇华吴仁洲程爱群
Owner 蒋建华
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