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Anther culture method of primula forbesii

A cultivation method and anther technology, applied in the field of plant tissue culture, can solve the problems of difficult genetic stability, long breeding cycle, time-consuming and labor-intensive, etc., and achieve good growth, less pests and diseases, and strong adaptability

Active Publication Date: 2010-07-07
BEIJING FORESTRY UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Primula forbesii Franch. is a typical self-incompatibility plant. Long-term cross-pollination makes the traits of offspring widely segregated and stable inheritance is difficult.
The traditional hybridization method is time-consuming and labor-intensive, and the breeding cycle is long

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Observation on Microspore Development Period of Primula forbesii Franch.

[0023] Flower buds of different sizes are collected during the flowering period, the length of the flower buds is measured respectively, and indicators such as the development period of the flower buds are observed. Use tweezers to peel off the sepals and petals, peel off the anthers and place them on a glass slide, add a small drop of carbo fuchsin, mash them with tweezers, remove the residue, cover with a cover glass, examine with a microscope, and summarize the length and appearance of the flower buds, etc. Relationship with microspore development stage. Through experimental observation, it was found that the microspore development of Primula forbesii Franch. can be divided into four stages, which are tetrad stage, uninucleate middle stage, uninucleate marginal stage and mature stage. The flower buds are not full and the length is less than 3.0 mm. At this time, the microspores enclosing the ...

Embodiment 2

[0025] The anthers whose microspore development period was the uninucleate marginal stage of Primula forbesii Franch. were used as explants for callus induction culture. To explore the optimal base medium for callus induction and the optimal ratio of hormone concentration.

[0026] At about 9 o'clock in the morning when the weather is fine and dry, collect the flower buds of the single-nuclei marginal stage grown in the greenhouse for future use. First soak in 70% alcohol for 30s, then disinfect with 2% sodium hypochlorite solution for 10 minutes, and rinse with sterile water for 3 to 4 times. Use tweezers to peel off the anthers on the ultra-clean workbench, remove the filaments, try to avoid the anthers from being damaged, inoculate in a petri dish, and seal it with a safety film. 20 anthers were inoculated per dish, and 5 dishes were inoculated per treatment.

[0027] Select MS, B 5 , N 6 The three kinds of media are used as the basic media for the culture of small prim...

Embodiment 3

[0030] Screening of Primula forbesii Franch. callus differentiation medium.

[0031] The proliferated callus was transferred to differentiation medium. The differentiation medium uses MS as the basic medium, 6-BA and NAA as the hormone species, and the concentration gradient of 6-BA is 0.1mg / L, 0.2mg / L, 0.5mg / L, 1.0mg / L, 2.0mg / L, the concentration gradient of NAA was 0.01mg / L, 0.1mg / L, 0.2mg / L, 0.5mg / L, and 10 kinds of differentiation medium were combined to explore their effects on the callus differentiation of Primula minor. 40g / L sucrose, 7g / L agar, pH 5.9-6.0. The lighting conditions are artificial auxiliary light 2000-3000Lux. The temperature is 24-26°C, and the light time is 12-14h / d.

[0032]40-60 days after the callus was inserted into the differentiation medium, axillary buds were differentiated on the differentiation medium of 6-BA 0.2mg / L+NAA 0.01mg / L, new leaves were continuously drawn out, and a large number of adventitious buds were continuously formed on the...

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Abstract

The invention provides an anther culture method of primula forbesii, comprising selection of the explant, disinfection and vaccination of the explant, induction and proliferation of the callus, differentiation of the callus, and rooting culture and transplantation of the tissue culture seedling, wherein, the medium of the induction and proliferation of the callus is as follows: MS +6- BA1.0mg / L +2,4-D 0.5mg / L, 30g / L sucrose, 6g / L agar, pH 5.9-6.0; the differentiation medium of the callus is as follows: MS +6- BA 0.2mg / L + NAA 0.01mg / L, 40g / L sucrose, 7g / L agar, pH 5.9-6.0; the rooting medium is as follows: MS, pH 5.9-6.0. The seedling obtained through the method provided by the invention has good growth, strong consistency and adaptability, less pests and diseases, easy management, and is easy for scale production.

Description

technical field [0001] The present invention relates to plant tissue culture, in particular to a method for cultivating anthers of Primula forbesii Franch. Background technique [0002] Small Primula (Primula forbesii Franch.) is a biennial herbaceous plant of Primulaceae Primulaceae unique to my country. It is native to the Yunnan region of my country at an altitude of 1500-2000m. Primula forbesii Franch. is a typical self-incompatibility plant. Long-term cross-pollination makes its progeny characters segregate widely and it is difficult to stably inherit them. The traditional hybridization method is time-consuming and laborious, and the breeding cycle is long. [0003] In order to shorten the breeding process, the excellent primula resources should be purified and stabilized as soon as possible. It is necessary to adopt the method of anther culture to obtain the haploid of Primula minor to form a complete set of anther culture technology system and lay the foundation for...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张启翔贾茵潘会堂孙明程堂仁董玲玲
Owner BEIJING FORESTRY UNIVERSITY
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