Method for rapidly obtaining haplobiont of Brassica oleracea var. botrytis

A purple flower and plant technology, which is applied in the field of plant tissue culture, can solve the problems of no reports on the application of purple lily microspore culture technology, and achieve the effects of operability, quality improvement, and application efficiency

Active Publication Date: 2014-12-10
天津天隆在田农业科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although scholars at home and abroad have done a lot of in-depth research on the application of microspore culture technology in cruciferous plants, there is no report on the application of microspore culture technology in purple lily.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The cultivation method is carried out as follows.

[0028] (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:

[0029] 1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH 6.0, high temperature and high pressure;

[0030] 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 2O 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.

[0031] 2) Embryoid body differentiation medium: B5 medium + sucrose 20g / L, agar 10g / L, pH 6.0, high temperature and high pressure sterilization;

[0032] 3) NLN-13 induction medium, NLN...

Embodiment 2

[0046] In addition to step (1) in 1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH 6.0, high temperature and high pressure; 2) NLN‐13 induction medium: NLN‐13 liquid medium 1L + sucrose 130g / 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;

[0047] (2) The culture method that co-cultivation of purple lily microspores and cabbage anthers promotes embryogenesis:

[0048] 1) Selection of flower buds: Take the flower buds with a petal-to-anther length ratio of 1.0 for purple cauliflower, 1.0 for cabbage, late mononucleate to early binucleate, healthy, and free from diseases and insect pests, as donors for microspore culture;

[0049] 2) Sterilization of flower buds: use 1g of mercuric liter + 1L of sterile water to prepare a sterili...

Embodiment 3

[0057]In addition to step (1) in 1) B5 washing medium: B5 liquid medium 1L + sucrose 30g / L, pH 6.0, high temperature and high pressure; 2) NLN‐13 induction medium: NLN‐13 liquid medium 1L + sucrose 130g / 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;

[0058] (2) The culture method that co-cultivation of purple lily microspores and cabbage anthers promotes embryogenesis:

[0059] 1) Selection of flower buds: take the flower buds with a petal-to-anther length ratio of 1.1 for purple cauliflower, 1.1 for cabbage, late mononucleate to early binucleate, healthy, and free from diseases and insect pests, as donors for microspore culture;

[0060] 2) Sterilization of flower buds: use 1g of mercuric liter + 1L of sterile water to prepare a steriliz...

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Abstract

The invention discloses a method for rapidly obtaining haplobiont of Brassica oleracea var. botrytis. The method comprises the following steps: sterilizing Brassica oleracea var. botrytis flower buds, then adding B5 wash medium to prepare a suspension liquid, filtering to obtain a filtrate, and centrifuging to obtain precipitate; sequentially adding an NLN-13 induction medium and activated carbon mixed solution to obtain microspore suspension liquid; subpackaging into a sterile culture dish, then adding sterile brassica oleracea anthers, 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, hardening seedlings and transplanting, to obtain regenerated plants. According to the method, the brassica oleracea anthers easy to form embryos and the Brassica oleracea var. botrytis isolated microspores difficult to form embryos are mixed for culture according to certain proportion, so that the embryonic development of the Brassica oleracea var. botrytis 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 method for cultivating naploid plants of Violet flower. Background technique [0002] Purple cauliflower (Brassica oleracea var.botrytis) belongs to the Brassica genus Brassica species of Brassicaceae. It is native to Western Europe and Italy. It is a variety of cauliflower. Other traits are similar to cauliflower. Purple cauliflower is rich in nutrition, rich in protein, vitamin C, and less in crude fiber. In addition, it also contains vitamins A, B1, B2, sucrose, fructose, selenium, etc. It is delicious in flavor, can improve human immune function, promote liver detoxification, and enhance human immunity. Physical fitness and disease resistance. The selenium contained in it can inhibit the growth of cancer cells. [0003] At present, most of the varieties of purple cauliflower used in production are imported from foreign countries, and the seeds are expensive,...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00C05G1/00
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 天津天隆在田农业科技有限公司
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