Differentiation culture medium adapted for indica rice isolated culture

A technology of differentiation medium and in vitro culture, applied in the field of new plants, can solve the problems of test failure, slow callus growth, unsatisfactory effect, etc., and achieve the effect of wide pertinence and applicability

Inactive Publication Date: 2008-12-24
HUAZHONG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, most of the reports on callus subculture and differentiation in the in vitro culture of indica rice varieties use MS or N 6 culture medium, but the results using such medium are very unsatisfactory
The main problem is that the growth rate of the existing subcultured callus is very slow, and the browning of the callus is serious, so it cannot be subcultured continuously, which leads to the failure of the whole experiment

Method used

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  • Differentiation culture medium adapted for indica rice isolated culture
  • Differentiation culture medium adapted for indica rice isolated culture
  • Differentiation culture medium adapted for indica rice isolated culture

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0020] Embodiment 1: Screening of indica rice callus subculture medium

[0021] The experimental design is divided into two parts: 1. Using MS, N 6 , B 5 and N 6 B (the medium is N 6 macronutrients and B 5 Trace and organic nutrients) based on the four most commonly used basic media (shown in Table 5), set 1 times (indicated by X, the same below), 2X, and combine with two carbon sources of sucrose and maltose to form 16 Seed medium: 2. Based on MS macronutrients and maltose as carbon source, set MS, N 6 , B 5 Trace elements, each trace element set 1X, 2X, 5X, 10X four water and a half, Fe 2+ Four gradients of 1X, 1.5X, 3X, and 5X were set up to form 48 culture media. A total of 64 designed media were inoculated with indica rice callus for subculture, and each medium was inoculated with 5 bottles of callus. Incubate in the dark at 26±1°C for 20 days. By observing and comparing the growth conditions (growth speed, texture and color) of the callus on various media, the s...

Embodiment 2

[0022] Embodiment 2: Comparative test of indica rice callus subculture effect

[0023] The test uses four representative indica rice varieties that are widely planted in China and have large genotype differences: Minghui 63 (a three-line indica restorer line), Zhenshan 97B (a three-line indica maintainer line) ), Zhong 419 (a three-line or two-line restorer line of indica rice) and W9864S (a two-line sterile line of indica rice). J of the present invention 3 The medium and the other three are commonly used basic medium MS (coded as J 1 ), N 6 (No. J 2 ) and B 5 (No. J 4 ) composed of subculture medium, the additional components added in these medium are the same as the amount of plant hormones such as 2,4-D, proline, glutamine, maltose and plant gum, the difference is only in the composition of the basic medium and Dosage, pH are 5.9. Do a comparison test of medium performance.

[0024] Experimental design of different subculture media for indica rice:

[0025] No. J ...

Embodiment 3

[0034] Embodiment 3: indica rice callus subculture medium J 3 Effect Test on Continuous Subculture of Indica Rice Callus

[0035] The callus of above-mentioned four kinds of indica rice varieties for testing provided by the embodiment is inserted into the described J 3 Culture medium, carry out continuous subculture test, the time of each subculture is 20 days. After each subculture, weigh and inoculate each 5 bottles of each test variety callus and do the differentiation activity measurement, to evaluate the J of the present invention 3 The ability of the culture medium to maintain calligenicity in continuous subculture. The test results are shown in Table 2.

[0036] Table 2 Indica rice callus subculture medium J 3 Determination of Differentiation Vitality of Indica Rice Callus Subcultured Continuously

[0037]

[0038] Note: a The total weight of calli inoculated into five bottles: b The total number of differentiated plants of callus in five bottles

[0039]The re...

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Abstract

The invention discloses a differentiation culture medium preparing method and application of rice, which comprises the following parts: 2800-3000mg / L KNO3,350-450mg / L (NH4)2SO4,300-400mg / L KH2PO4, 180-200mg / L MgSO4 .7H2, 170-200mg / L CaCl2 .2H2, 45-55mg / L FeSO4 .7H2O, Na2EDTA 60-74mg / , 80-100mg / L MnSO4 .4H2, 15-25mg / L ZnSO4 .7H2O, 25-30mg / L H3BO,6.0-8.0mg / L KI, 0.2-0.3mg / L CuSO4 .5H2O,2.0-3.0mg / L Na2MoO4 .2H2O, 0.2-0.3mg / L CoCl2 .6H2O,2.0-3.0mg / L aminoacetic acid, 0.5-1.0mg / L VB1, 1.0-1.5mg / L VB6,1.0-1.5mg / L niacin,100-150mg / L inositol,300-500mg / L glutamine,500-800mg / L proline,30-50g / L maltose,2mg / L 6-aminopurine benzyl,2mg / L activator,0.2mg / L heteroauxin,0.2mg / L 1-naphthalene acetic acid,5g / L vegetable glue with pH value at 6.0.

Description

[0001] This application is a divisional application of the patent application No. 200410013345.7 submitted on June 24, 2004. technical field [0002] The invention belongs to the technical field of new plants. It specifically relates to an in vitro culture method of indica rice, and more specifically relates to the improvement of indica rice callus subculture and differentiation medium and its application in indica rice in vitro culture and transgene. Background technique [0003] The culture medium is the key to the plant in vitro culture method, and the establishment of any plant in vitro culture method is mainly carried out around the screening of the culture medium. Plant in vitro culture is the process of artificially simulating the natural growth of plants indoors. Different species have different growth conditions and environments. Therefore, it is conceivable that it is impossible to design a medium suitable for tissue culture of all species. Each medium Each has it...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N5/04
Inventor 林拥军张启发
Owner HUAZHONG AGRI UNIV
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