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In vitro preservation culture medium for cassava embryogenic callus

A technology of embryogenic callus and in vitro preservation, which is applied in the field of agricultural biology, can solve the problems of high energy consumption, genetic variation, and in vivo preservation in nurseries are vulnerable to invasion by diseases and insect pests, so as to reduce the possibility of genetic variation and maintain traits Effect

Inactive Publication Date: 2016-01-13
SOUTH ASIAN TROPICAL AGRI SCI RES INST OF GUANGXI
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  • Application Information

AI Technical Summary

Problems solved by technology

However, the preservation of living organisms in nurseries is vulnerable to the invasion of diseases and insect pests and the threat of extreme environments; for the preservation of test-tube seedlings, long-term in vitro culture requires regular subculture, and may cause genetic variation; low-temperature or ultra-low temperature in vitro preservation equipment is costly and energy-intensive high

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  • In vitro preservation culture medium for cassava embryogenic callus

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0025] Prepare the improved GD medium, wherein the components and the contents of each component per liter of the medium are as follows:

[0026] 1. Major elements: Ca(NO 3 ) 2 2H 2 O230mg / L; KCl70mg / L; KH 2 PO 4 350mg / L;

[0027] KNO 3 1100mg / L; MgSO 4 18mg / L; NH 4 NO 3 1100mg / L;

[0028] 2. Trace elements: CaCl 2 ·6H 2 O0.037mg / L; CuS0 4 ·5H 2 O0.037mg / L; FeNaEDTA38.5mg / L; H 3 BO 3 0.40mg / L; KI0.90mg / L; MnSO 4 ·H 2 O1.25mg / L; Na 2 MoO 4 2H 2 O0.027mg / L; ZnSO 4 ·7H 2 O0.40mg / L;

[0029] 3. Growth inhibitors: TIBA6mg / L-8mg / ;

[0030] 4. Phytohormone: 2,4-D0.7mg / L;

[0031] 5. Others: sucrose 22g / L; agarose 6.5g / L; water; pH 5.8

[0032] 6. Culture medium preparation: Add according to the content of each component, in the order of macroelements → trace elements → plant hormones → sucrose → a mixture of boiled agar and water → add water to make up the volume → adjust the pH value to 5.8 → divide into petri dishes Neutralize and seal well → autoclave ste...

Embodiment 2

[0047] Configure cassava embryogenic callus in vitro preservation medium, wherein the components and the contents of each component of each liter of the medium are as follows:

[0048] 1. Major elements: Ca(NO 3 ) 2 2H 2 O210mg / L; KCl65mg / L; KH 2 PO 4 300mg / L;

[0049] KNO 3 1000mg / L; MgSO 4 17mg / L; NH 4 NO 3 1000mg / L;

[0050] 2. Trace elements: CaCl 2 ·6H 2 O0.025mg / L; CuS0 4 ·5H 2 O0.025mg / L; FeNaEDTA37.0mg / L; H 3 BO 3 0.30mg / L; KI0.80mg / L; MnSO 4 ·H 2 O1.00mg / L; Na 2 MoO 4 2H 2 O0.025mg / L; ZnSO 4 ·7H 2 O0.30mg / L;

[0051] 3. Growth inhibitors: TIBA7mg / ;

[0052] 4. Phytohormone: 2,4-D0.7mg / L;

[0053] 5. Others: sucrose 20g / L; agarose 6.0g / L; water; pH 5.5

[0054] Culture medium preparation: add according to the content of each component, in the order of macroelements→trace elements→phytohormones→growth inhibitors→sucrose→mixture of boiled agar and water→add water to volume→adjust pH value to 5.5→pack to Put it in a petri dish and seal it well→st...

Embodiment 3

[0058] Configure cassava embryogenic callus in vitro preservation medium, wherein the components and the contents of each component of each liter of the medium are as follows:

[0059] 1. Major elements: Ca(NO 3 ) 2 2H 2 O250mg / L; KCl75mg / L; KH 2 PO 4 400mg / L;

[0060] KNO 3 1200mg / L; MgSO 4 20mg / L; NH 4 NO 3 1200mg / L;

[0061] 2. Trace elements: CaCl 2 ·6H 2 O0.050mg / L; CuS0 4 ·5H 2 O0.050mg / L; FeNaEDTA40.0mg / L; H 3 BO 3 0.50mg / L; KI1.00mg / L; MnSO 4 ·H 2 O1.50mg / L; Na 2 MoO 4 2H 2 O0.030mg / L; ZnSO 4 ·7H 2 O0.50mg / L;

[0062] 3. Growth inhibitors: TIBA6mg / ;

[0063] 4. Phytohormone: 2,4-D0.7mg / L;

[0064] 5. Others: sucrose 25g / L; agarose 6.5g / L; water; pH 6.0

[0065] Culture medium preparation: add according to the content of each component, in the order of macroelements→trace elements→phytohormones→growth inhibitors→sucrose→mixture of boiled agar and water→add water to make up to volume→adjust pH value to 6.0→pack to Put it in a petri dish and seal...

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Abstract

The invention discloses an in vitro preservation culture medium for cassava embryogenic callus. A formula of the in vitro preservation culture medium is that an improved GD culture medium is added with 0.6-0.8mg / L of 2,4-D, 6-7mg / L of LTIBA, 20-25g / L of saccharose and 6.0-6.5g / L of agar. The in vitro preservation culture medium is placed under conditions with a temperature of 13-17 DEG C, illumination of 100-1200Lx and illumination time of 16-18h / d to perform culturing. The culture medium is adopted to carry out in vitro preservation on the cassava embryogenic callus, has embryogenic callus multiplication amount which is 1.0-1.2 times that of a traditional culture medium, cell activity which is 2-4 times that of the traditional culture medium and the best preserving effect when subculture time is prolonged to 120d. The method is adopted to carry out in vitro preservation for cassava embryogenic callus, so that basis is provided for storing cassava germplasm resources.

Description

technical field [0001] The invention belongs to the field of agricultural biotechnology, and in particular relates to a method for in vitro preservation of cassava embryogenic callus. Background technique [0002] Cassava (manihotesculenta Crantz) is a shrub-like plant of the Euphorbiaceae (Euphorbiaceae) cassava genus (Manihot). One of the potato crops, known as "underground granary" and "starch king", it is widely cultivated in tropical and subtropical regions, and is widely cultivated in my country. The cultivation, processing and utilization of cassava occupy an important position in agricultural production . [0003] Research on Tropical Crops Variety Resources of Chinese Academy of Tropical Agricultural Sciences Through the investigation of domestic cassava germplasm resources, a batch of cassava germplasm resources at home and abroad have been collected, becoming the only national cassava germplasm resource nursery in my country, and cassava seed Quality resources in v...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 黎萍刘连军李恒锐陈海生谢君锋何洪良农秋莲邱文武杨海霞青鑫韦雪英符策冯兰
Owner SOUTH ASIAN TROPICAL AGRI SCI RES INST OF GUANGXI
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