Tissue culture method of monocotyledons

A technique for monocotyledonous plants and seeds, which is applied in the field of tissue culture of monocotyledonous plants, can solve the problems of long screening period, high difficulty, and heavy workload, and achieve the effects of reducing the difficulty of selection, easy operation, saving time and cost

Inactive Publication Date: 2014-05-07
CHINA AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Screening and breeding switchgrass materials with high regeneration ability may be an effective method, but for most switchgrass varieties, it is difficult, heavy workload, and long selection cycle to obtain new lines with high regeneration ability, and at least 2-3 rounds (years) of selection are required Only in this way can it be possible to obtain highly efficient recycled materials

Method used

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  • Tissue culture method of monocotyledons
  • Tissue culture method of monocotyledons
  • Tissue culture method of monocotyledons

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0068] Embodiment 1, the acquisition and tissue culture of type II callus of switchgrass variety Alamo

[0069] Test material: mature seeds of switchgrass variety Alamo. Alamo is a lowland switchgrass with high biomass, flowering in autumn, moderate salt tolerance, suitable for most soil types, and is a common material for tissue culture of switchgrass. The tested Alamo mature seeds were produced in Texas, USA (25°50'N-36°30'N) in 2010.

[0070] (1) Seed disinfection

[0071] Select plump seeds of Alamo, sterilize with 5% (volume fraction) sodium hypochlorite aqueous solution for 1.5 h, wash with sterile deionized water 5 times, soak in sterile deionized water and place in an incubator at 4 °C for 3 days. After 3 days, it was treated with 5% (volume fraction) sodium hypochlorite aqueous solution for 30 minutes, and washed 5 times with sterile deionized water.

[0072] (2) Induction of callus

[0073] During callus induction, the following three media were selected as callu...

Embodiment 2

[0118] Example 2, the acquisition and tissue culture of type II callus of switchgrass variety Performer

[0119] Test material: mature seeds of switchgrass variety Performer. Performer is a lowland type switchgrass with high nutritional value, excellent forage quality and high biomass. It is a common material for switchgrass tissue culture. The mature seeds of Performer tested were produced in North Carolina, USA (33°50N-36°35'N) in 2010.

[0120] (1) Seed disinfection

[0121] Refer to step (1) of Example 1 for operation.

[0122] (2) Induction of callus

[0123] Refer to step (2) of Example 1 for operation.

[0124] The callus induction rate is shown in Table 5. For treatment ① (direct induction), the callus induction rate on MB solid medium was 53.34%, the callus induction rate on MS7 solid medium was 41.11%, and the callus induction rate on NB7 solid medium The callus induction rate on the medium was 52.78%, and there was no significant difference in the callus induc...

Embodiment 3

[0148] Example 3, the acquisition and tissue culture of type II callus of switchgrass variety Carthage

[0149] Test material: mature seeds of switchgrass variety Carthage. Carthage is a transitional switchgrass variety with many leaves, fast-growing stalks, and green in early spring. Its biomass exceeds that of Midwestern varieties in the United States and has high nutritional value. Due to difficult regeneration, it is rarely used in tissue culture. The mature seeds of Carthage tested were produced in North Carolina (33°50'N-36°35'N) in 2010.

[0150] (1) Seed disinfection

[0151] Refer to step (1) of Example 1 for operation.

[0152] (2) Induction of callus

[0153] Refer to step (2) of Example 1 for operation.

[0154] The callus induction rate is shown in Table 9. For treatment ① (direct induction), the callus induction rate on MB solid medium was 60.56%, the callus induction rate on MS7 solid medium was 61.67%, and the callus induction rate on NB7 solid medium The ...

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Abstract

The invention discloses a tissue culture method of monocotyledons. The method relates to a method for obtaining an II type callus of the monocotyledons, comprising the following steps: 1, with mature seeds of monocotyledons as explants, inoculating the explants on a callus-inducing culture medium, induction-culturing for 4-6 weeks at temperature of 25+ / -2 DEG C under the condition of light resistance to obtain a callus 1; 2, peeling a white callus at the outermost layer from the callus 1 obtained from the step 1, collecting an inside yellow embryonic callus, inoculating the inside yellow embryonic callus on a callus subculture medium, culturing for 3 weeks at the temperature of 25+ / -2 DEG C under the condition of light resistance to obtain a callus 2; and 3, selecting a callus the volume of which is increased by above 60 percent within 3 weeks of subculturing from the callus 2 obtained from the step 2, namely, the II type callus. Experiments prove that the tissue culture method is suitable for switchgrasses of different ecotypes, and is capable of obtaining an excellent callus with high regeneration efficiency within short time, thus the time and cost are saved, and the biological breeding process of switchgrass genetic modification is accelerated.

Description

technical field [0001] The invention belongs to the field of plant tissue culture, and relates to a method for tissue culture of monocot plants. Background technique [0002] Cell regeneration in monocots mostly occurs through somatic embryogenesis. Efficient and fast access to high-frequency embryogenic callus is the basis for biotechnology breeding of monocotyledonous plants. However, for most inbred lines of monocotyledonous plants such as switchgrass, wheat, and maize, it takes a long time for callus culture to obtain embryogenic cell lines that can regenerate efficiently and can also undergo genetic transformation. [0003] Switchgrass (Panicum virgatum L.) belongs to the genus Panicum virgatum of the Poaceae (Gramineae) and is a perennial herb native to the North American continent. It has strong adaptability, low requirements on the soil environment, and large biomass. energy plant. However, due to the high lignin content of switchgrass, the stress resistance needs...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 张万军刘燕蓉马宁叶文兴张蕴薇
Owner CHINA AGRI UNIV
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