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Method and special culture medium for regenerated seedlings of luffa unfertilized ovary embryoids

A technology of embryoid body and culture medium, which is applied in the field of seedling regeneration method and special medium of loofah unfertilized ovary embryoid body, which can solve the problem of low occurrence frequency, haploid induction of loofah gynogenesis, Difficult to apply and other problems, to achieve the effect of improving the seedling rate

Inactive Publication Date: 2019-10-08
杨泗海
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The former occurs very infrequently and is difficult to apply
However, there is no research report on loofah inducing haploid through gynogenesis through unfertilized ovary or ovule

Method used

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  • Method and special culture medium for regenerated seedlings of luffa unfertilized ovary embryoids
  • Method and special culture medium for regenerated seedlings of luffa unfertilized ovary embryoids
  • Method and special culture medium for regenerated seedlings of luffa unfertilized ovary embryoids

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] 1 Materials and methods

[0025] 1.1 Test materials:

[0026] The embryoid bodies formed by induction and culture of unfertilized ovaries of common loofah varieties 'Nanjing Snake-shaped Loofah' and 'Wanlu No. 1' were obtained from the inventor's previous experiments.

[0027] 1.2 Test method:

[0028] (1) Low-temperature pretreatment of embryoid bodies: the embryoid bodies produced by in vitro induction culture of unfertilized ovaries of loofah were placed in a 4°C refrigerator for low-temperature pretreatment for 2-3 days.

[0029] (2) Embryoid body seedling cultivation: transfer the pretreated embryoid body to the seedling growth medium at a temperature of 25°C, light and dark culture time of 16h / d and 8h / d respectively, light intensity Cultivate in a culture room of 2800 lx, and make the embryoid body differentiate into complete test-tube plantlets with leaves and roots.

[0030] The formula of the basic seedling medium is: NH 4 NO 3 2025mg / L, KNO 3 1900mg / L, ...

Embodiment 2

[0049] According to the research results in Example 1, the best seedling medium of the present invention is: NH 4 NO 3 2025mg / L, KNO 3 1900mg / L, NaH 2 PO 4 ·H 2 O 150mg / L, calcium citrate 340mg / L, MgSO 4 ·7H 2 O 375mg / L, KI 0.83mg / L, H 3 BO 3 5.8mg / L, MnSO 4 4H 2 O 10mg / L, ZnSO 4 ·7H 2 O 2.5mg / L, Na 2 MoO 4 2H 2 O 0.25mg / L, CuSO 4 ·5H 2 O 0.025mg / L, CoCl 2 ·6H 2 O 0.025mg / L, ferrous lactate 31.6mg / L, inositol 100mg / L, niacin 0.9mg / L, pyridoxine hydrochloride 1.0mg / L, thiamine hydrochloride 5.9mg / L, glycine 2.7mg / L , lysine 4.1mg / L, GA 30.5mg / L, 5-nitroguaiacol sodium 0.05mg / L, 5-aminolevulinic acid 1.0mg / L, sucrose 30g / L, yeast extract powder 280mg / L, polyvinyl alcohol 1.0g / L, Praseodymium sulfate 2.0mg / L, tomato juice 70ml / L, vegetable gel 12g / L.

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Abstract

The invention discloses a method and special culture medium for regenerated seedlings of luffa unfertilized ovary embroyoids. The special culture medium comprises a large number of elements such as NH4NO3 and KNO3, trace elements such as MnSO4 4H2O, ferrous lactate, inositol, niacin, pyridoxine hydrochloride, thiamine hydrochloride, glycine, lysine, GA3, 5-nitroguaiacol sodium, 5-aminoethanolpionic acid, saccharose, yeast extract powder, polyvinyl alcohol, praseodymium sulfate, tomato juice and plant gel. The method includes the steps of low temperature pretreatment of the embryoids, seedlingculture of the embryoids, seeding refining of regenerated plants and transplanting of the regenerated plants. Ploidy identification is carried out through a flow cytometry, and the average haplotype rate of embryo sac regenerated plants is 16.3%; the luffa haploid pants are obtained successfully, which proves that the method of obtaining haploids in vitro culture of luffa unfertilized ovary embroyoids is feasible, and opens up a new way for haploid breeding of luffa.

Description

technical field [0001] The invention belongs to the field of biotechnology, and in particular relates to a seedling regeneration method and a special culture medium for loofah unfertilized ovary embryoid bodies. Background technique [0002] Loofah belongs to Cucurbitaceae ( Cucurbitaceae ) Luffa ( Luffa spp .) Annual climbing herb, native to East India, mainly distributed in tropical and subtropical Asia, cultivated in both north and south of my country, and is the main melon vegetable in my country. Loofah has two cultivars of common loofah and ribbed loofah in my country. Ribbed loofah is mainly cultivated in Guangdong, Guangxi, Hainan and other places, and ordinary loofah is mainly cultivated in other areas. Loofah is a short-day crop, which likes strong sunlight and is more resistant to weak light. Especially ribbed loofah is a typical short-day plant, which has strict requirements on short-day sunshine and grows slowly under long-day conditions. Loofah is resistant...

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 杨泗海
Owner 杨泗海
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