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Method for improving embryogenesis rate of Raphanus sativus L. sinoruber makino

A rouge radish and occurrence rate technology, applied in the field of plant tissue culture, can solve the problems of hindering the application of the microspore culture technology, the microspore culture technology has not been applied, the embryoid body induction rate and the regeneration rate are low, and the method is simple and easy to achieve. performance, improve application efficiency, and reduce workload

Active Publication Date: 2014-12-10
南京利华农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

After years of hard work, although scholars at home and abroad have done a lot of in-depth research on improving the frequency of radish microspore embryogenesis, compared with other cruciferous crops, its embryoid body induction rate and regeneration rate are low, which seriously hinders the development of microspore embryos. The application of microspore culture technology
However, in the breeding of rouge radish, microspore culture technology has not been applied

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0028] The cultivation method is carried out as follows.

[0029] (1) Culture medium preparation: comprise the culture medium of microspore different culture stages, its component and the weight that each component contains in every liter of medium are:

[0030]1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH 6.0, high temperature and high pressure;

[0031] The B5 liquid medium, in 1L, consists of: NaH 2 PO 4 2H 2 O 169.5mg, KNO 3 2500mg, (NH 4 ) 2 SO 4 134mg, MgSO 4 ·7H 2 O 500mg, MnSO 4 4H 2 O 10mg, H 3 BO 3 3mg, ZnSO 4 7H2O 2mg, KI 0.75mg, Na 2 MoO 4 2H 2 O 0.25mg, CuSO 4 ·5H 2 O 0.025mg, CoCl 2 ·6H 2 O 0.025mg, Na 2 -EDTA 37.3mg, FeSO 4 ·7H 2 O 27.8mg, CaCl 2 .2H 2 O 150mg, VB1 10mg, VB6 1mg, VPP 1mg, inositol 100mg and the rest distilled water.

[0032] 2) Embryoid body differentiation medium: B5 medium + sucrose 20g / L, agar 10g / L, pH 6.0, high temperature and high pressure sterilization;

[0033] 3) NLN-13 induction medium, NLN ...

Embodiment 2

[0047] In addition to step (1): 1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH6.0, high temperature and high pressure; 2) NLN-13 induction medium: NLN-13 liquid medium 1L + sucrose 130g / L L, pH 6.1, filter sterilized; 3) Embryoid body differentiation medium: B5 medium + sucrose 20g / L, agar 11g / L, pH 6.0, high temperature and high pressure sterilization; 4) Rooting medium: MS medium + White sugar 20g / L, agar 7g / L, pH 5.9, high temperature and high humidity sterilization;

[0048] (2) Co-cultivation of rouge radish microspores and broccoli anthers to promote embryogenesis:

[0049] 1) Selection of flower buds: the petal-to-anther length ratio of rouge radish is 1.0, the broccoli is 1.0, the late mononucleate to early binucleate, healthy, and free from diseases and insect pests are used as donors for microspore culture;

[0050] 2) Sterilization of flower buds: use 1g of mercury liter + 1L of sterile water to prepare a sterilizing solution; put the mixed flower bu...

Embodiment 3

[0058] In addition to step (1): 1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH6.0, high temperature and high pressure; 2) NLN-13 induction medium: NLN-13 liquid medium 1L + sucrose 130g / L L, pH 6.1, filter sterilized; 3) Embryoid body differentiation medium: B5 medium + sucrose 20g / L, agar 11g / L, pH 6.0, high temperature and high pressure sterilization; 4) Rooting medium: MS medium + White sugar 20g / L, agar 7g / L, pH 5.9, high temperature and high humidity sterilization;

[0059] (2) Co-cultivation of rouge radish microspores and broccoli anthers to promote embryogenesis:

[0060] 1) Selection of flower buds: the petal-to-anther length ratio of rouge radish is 0.9, the broccoli is 0.9, the late mononucleate to early binucleate, healthy, and free from diseases and insect pests are taken as donors for microspore culture;

[0061] 2) Sterilization of flower buds: use 1g of mercury liter + 1L of sterile water to prepare a sterilizing solution; put the mixed flower b...

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Abstract

The invention discloses a method for improving the embryogenesis rate of Raphanus sativus L. sinoruber makino. The method comprises the following steps: sterilizing Raphanus sativus L. sinoruber makino flower buds, then adding B5 wash medium to prepare a suspension liquid, filtering to obtain a filtrate, and centrifuging to obtain precipitate; sequentially adding NLN-13 induction medium and an activated carbon mixed solution to obtain microspore suspension liquid; subpackaging into a sterile culture dish, then adding sterile broccoli anther, performing Parafilm sealing and then conducting heat-shock treatment; performing routine culture to obtain cotyledonous embryoid; inoculating to an embryoid differentiation medium until differentiation is performed, and regenerating buds; cutting and inoculating regenerated buds to a rooting medium for rooting culture, and hardening seedling and transplanting, to obtain regenerated plants. According to the method, the broccoli anther easy to form embryos and the Raphanus sativus L. sinoruber makino isolated microspore difficult to form embryos are mixed for culture according to certain proportion, so that the embryonic development of the Raphanus sativus L. sinoruber makino can be promoted to obtain a great amount of regenerated plants, and the culture efficiency can be improved.

Description

technical field [0001] The invention relates to the technical field of plant tissue culture, in particular to a culture method for promoting carmine radish microspore embryogenesis. Background technique [0002] Rouge radish (Raphanus Sativus L.Sinoruber Makino) is a local variety of radish in the genus Brassicaceae. Its fleshy root is rich in nutrition and denser in texture. It is suitable for processing various dried radishes. At the same time, the fleshy root bark and heart are all red. Radish has a high content of red pigment and its pigment quality and solubility are superior to other carrots. Therefore, red radish is an ideal raw material crop for extracting natural edible red pigment, and has a large cultivation area in Southwest my country. The radish red pigment extracted from rouge radish has the characteristics of good chemical stability, storage stability, and excellent heat resistance. In addition, it also has certain antioxidant properties and is a coloring agen...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 张振超毛忠良潘跃平吴国平王建华戴忠良秦文斌姚悦梅潘永飞肖燕孙春青马志虎孙国胜
Owner 南京利华农业科技有限公司
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