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Culture method for brassica oleracea L. var. acephala microspore regeneration plant

A microspore culture and kale technology, which is applied in the field of plant tissue culture, can solve the problems of affecting the embryo production rate, affecting the quality of the embryo, inhibiting the microspore cultivation and breeding technology, etc., and achieve the effect of increasing the embryo emergence rate and improving utilization efficiency

Inactive Publication Date: 2011-11-02
ZHEJIANG UNIV +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

This patented new technique allows for easier creation of hybrid crops with different types of vegetables such as carrots or cauliflower flowers by mixing them together without damaging their growth. By doing this it increases the yield from these plants compared to traditional methods like crop rotation.

Problems solved by technology

This patented describes various technical solutions aimed towards increasing the speed and yield of creation of edible crop species called brassic acid tamarina varies by growing them over several generations or developing new ones through crossing techniques such as seedling propagating technique. However, these conventional ways require significant labor input during each generation cycle due to their slow growth rates compared to natural occurrences found throughout this world's tropics. To address this issue, we propose a novel way to increase the germinancy rate of certain types of brassiercane grown commercially without requiring excessive human effort.

Method used

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  • Culture method for brassica oleracea L. var. acephala microspore regeneration plant
  • Culture method for brassica oleracea L. var. acephala microspore regeneration plant

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Experimental program
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Effect test

Embodiment 1

[0037] The cultivation method is carried out as follows.

[0038] (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:

[0039] Table 1 NLN and MS medium composition list

[0040]

[0041]

[0042] 1) NLN-13 induction medium: NLN liquid medium 1L + sucrose 130g / L, pH6.0, filter sterilized;

[0043] 2) Embryoid body differentiation medium: MS medium + sucrose 20g / L, agar 10g / L, pH 6.0, high temperature and high humidity sterilization;

[0044] 3) Rooting medium: MS medium + sucrose 30g / L, agar 6g / L, pH5.8, high temperature and high humidity sterilization;

[0045] (2) Cultivation of the regenerated plants of microspores of kale with high germination rate:

[0046] 1) Selection of flower buds of donor plants: take healthy and pest-free inflorescences of kale and rapeseed as donor plants for microspore culture; after induction at a low te...

Embodiment 2

[0056] In addition to step (1) in 1) NLN-13 induction medium: NLN-13 liquid medium 1L + sucrose 130g / L, pH6.1, filter sterilization; 2) embryoid differentiation medium: MS medium + sucrose 20g / L, agar 11g / L, pH6.1, high temperature and high humidity sterilization; 3) Rooting medium: MS medium + white sugar 20g / L, agar 7g / L, pH5.9, high temperature and high humidity sterilization;

[0057] (2) Cultivation of the regenerated plants of microspores of kale with high germination rate:

[0058] 1) Selection of flower buds of donor plants: take healthy and pest-free inflorescences of kale and rapeseed as donor plants for microspore culture; after induction for 48 hours at a low temperature of 3°C, the length of petals and anthers on the inflorescences are taken at a ratio of 1.1. Flower buds from late mononucleate to early binucleate;

[0059] 2) Sterilization of flower buds: use 5.6% (mass percentage concentration) sodium hypochlorite aqueous solution 56ml / L+ absolute ethanol 100m...

Embodiment 3

[0068] In addition to step (1) in 1) NLN-13 induction medium: NLN-13 liquid medium 1L + sucrose 130g / L, pH6.2, filter sterilization; 2) embryoid differentiation medium: MS medium + sucrose 20g / L, agar 12g / L, pH6.0, high temperature and high humidity sterilization; 3) rooting medium: MS medium + sucrose 25g / L, agar 6.5g / L, pH6.0, high temperature and high humidity sterilization;

[0069] (2) Cultivation of the regenerated plants of microspores of kale with high germination rate:

[0070] 1) Selection of flower buds of donor plants: take the inflorescences of kale and rapeseed that grow healthily and are free of diseases and insect pests as donor plants for microspore culture; take flower buds with a petal to anther length ratio of 0.8 on the inflorescence, late mononucleate to early binucleate;

[0071] 2) Sterilization of flower buds: use 5.6% (mass percentage concentration) sodium hypochlorite aqueous solution 56ml / L+dehydrated alcohol 100ml / L+clean liquid 9 drops+sterile wa...

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Abstract

The invention discloses a culture method for a brassica oleracea L. var. acephala microspore regeneration plant. The method comprises the following steps: taking inflorescences of brassica oleracea L. var. acephala and rape, directly taking or taking after induction alabastrum of the inflorescences, adding the alabastrum into NLN-13 induced medium after disinfection to form fluid suspension, carrying out filtering, and centrifuging the filtrate to obtain a precipitate; adding NLN-13 induced medium and active carbon mixed liquor in order so as to obtain a microspore fluid suspension; carrying out a heat shock treatment, followed by culturing so as to obtain embryoids in cotyledon stage; inoculating the embryoids to embryoid differential medium until the embryoids are differentiated and regenerates into buds; cutting the regenerated buds to a rooting medium for rooting culture, and hardening and transplanting seedlings to obtain regenerated plants; taking young leaves of the regenerated plants for the detection of ploidy of corresponding regenerated plants and determining regenerated seedlings of rape and brassica oleracea L. var. acephala in the regenerated plants. According to the method provided in the invention, rape which is easy to generate embryos and brassica oleracea L. var. acephala which is difficult to generate embryos are mixedly cultured; the material which is easy to generate embryos is used to spur the material which is difficult to generate embryos; therefore the ratio of embryos of brassica oleracea L. var. acephala is improved.

Description

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Claims

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Application Information

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Owner ZHEJIANG UNIV
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