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A highly efficient mutagenesis method for tobacco pollen

A kind of pollen and high-efficiency technology, applied in horticultural methods, botanical equipment and methods, applications, etc., can solve the problems of difficult absorption of mutagens, small variation range, and difficult screening, so as to overcome obvious and recessive obstacles and mutations The effect of large range and saving breeding time

Inactive Publication Date: 2011-12-28
TOBACCO RES INST OF HUBEI PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, these mutagenesis treatments have problems such as difficult absorption of mutagen, low frequency of mutagenesis, small variation range, difficulty in offspring screening, low yield and easy generation of chimeras, etc.

Method used

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  • A highly efficient mutagenesis method for tobacco pollen
  • A highly efficient mutagenesis method for tobacco pollen
  • A highly efficient mutagenesis method for tobacco pollen

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Embodiment 1: Flue-cured tobacco material 1 pollen in vitro liquid culture mutagenesis, the operation steps are as follows:

[0023] (1) Sample collection: Take healthy age-appropriate flower buds at the single-nuclei margin stage on the main inflorescence of the flue-cured tobacco plant, with a length range of 2.0-2.2 cm, and quickly store the taken flower buds in an ice box at 4-7°C;

[0024] (2), disinfection treatment: take out the flower buds in the ice box in step 1, carefully peel off the anthers in the flower buds with tweezers on the ultra-clean workbench, pack the anthers with sterile gauze and immerse them in 70% ethanol for 15 seconds, and then immerse In 10% saturated calcium hypochlorite solution for 8 minutes, rinse with sterile water 3 times, 5 minutes each time;

[0025] (3), collect pollen: put the anther after disinfection in the step 2 in the large test tube of round bottom, add the B5-13 liquid medium of 2ml (sucrose concentration is 13% in the B5 b...

Embodiment 2

[0032] Embodiment 2, flue-cured tobacco material 2 pollen isolated liquid culture mutagenesis, its operation steps are as follows:

[0033] (1) Sample collection: take healthy age-appropriate flower buds on the side inflorescences of flue-cured tobacco plants at the single-nuclei edge stage, with a length range of 2.1-2.3 cm, and quickly store the taken flower buds in an ice box at 4-7°C;

[0034](2), disinfection treatment: take out the flower buds in the ice box in step 1, carefully peel off the anthers in the flower buds on the ultra-clean workbench, pack the anthers with sterile commercial stockings and immerse them in 75% ethanol for 10 seconds, then Immerse in 10% saturated calcium hypochlorite solution for 10 minutes, rinse with sterile water twice, 5 minutes each time;

[0035] (3), collect pollen: move the anther after disinfection in step 2 in the big test tube of round bottom, add the B5-13 liquid medium of 3ml (sucrose concentration is 13% in the B5 basic medium), ...

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Abstract

The invention relates to a high-efficiency mutagenesis method of nicotiana tabacum L. pollen. The high-efficiency mutagenesis method comprises the following steps of putting nicotiana tabacum L. pollen into a liquid medium under an aseptic condition to carry out mutagenesis treatment by 0.1 to 0.2% of ethyl methanesulfonate (EMS) in a dark place at a temperature of 30 to 32 DEG C, carrying out chromosome doubling treatment by a NLN-16 liquid medium in a dark environment at a temperature of 30 to 32 DEG C, putting the nicotiana tabacum L. pollen obtained by the previous step into a NLN-13 liquid medium to carry out embryo induction culture in a dark environment at a temperature of 24 to 26 DEG C, carrying out shake culture by a shaking table in a dark environment at a temperature of 24 to 26 DEG C when an immature embryo appears, and putting the immature embryo growing a cotyledon into a solid medium to carry out subculture, wherein during a transplanting season, after seedling hardening, a seedling obtained by the high-efficiency mutagenesis method can be transplanted in a field for mutant character identification. The high-efficiency mutagenesis method overcomes dominant and recessive barriers of gene expression, is beneficial for selection, shortens greatly a mutant character stabilization period, saves breeding generation time, and has the characteristics of wide variation range, high mutagenesis frequency and the like.

Description

technical field [0001] The invention relates to the technical field of tobacco mutagenesis, in particular to an efficient mutagenesis method for tobacco pollen. Background technique [0002] Tobacco (Nicotiana tabacum L,.) belongs to Dicotyledoneae, Tubifiorae, Solanaceae Nicotiana in plant taxonomy, and is an important economic crop in many countries, among which flue-cured tobacco (2n=24II) is the type of tobacco with the largest planting area in China and in the world. [0003] For a long time, due to the excessive use of a few main parents in tobacco breeding, combined with directional selection, the genetic basis of bred varieties is narrow and the phenomenon of single varieties is becoming more and more serious, making it difficult to breed new varieties with greater breakthroughs. The embarrassing situation of "reviewing but not planting" of new varieties often occurs, which also brings great difficulties to the construction of molecular marker genetic linkage maps a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 蔡长春林国平曹景林王毅杨树张俊杰
Owner TOBACCO RES INST OF HUBEI PROVINCE
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