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A kind of efficient method for in vitro liquid culture of tobacco pollen

A liquid culture and pollen technology, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of low doubling efficiency, long anther germination time, heavy workload, etc., and achieve short pollen germination time and large Practical value, double the effect of high efficiency

Inactive Publication Date: 2011-12-21
TOBACCO RES INST OF HUBEI PROVINCE
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] Tobacco tissue culture was carried out earlier, mainly including pollen culture, anther culture and explant callus induction, etc. At present, anther culture has become the main means of tobacco tissue culture, but anther culture has long anther germination time and low doubling efficiency. Low, heavy workload in the later stage, interference of anther wall somatic callus, slightly low genetic homology of offspring populations, and difficulty in obtaining a large number of breakthrough breeding materials, and it is difficult to obtain a large number of target materials for plants with less pollen

Method used

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  • A kind of efficient method for in vitro liquid culture of tobacco pollen
  • A kind of efficient method for in vitro liquid culture of tobacco pollen
  • A kind of efficient method for in vitro liquid culture of tobacco pollen

Examples

Experimental program
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Effect test

Embodiment 1

[0027] Embodiment 1, the isolated liquid culture of flue-cured tobacco pollen, concrete steps are as follows successively:

[0028] (1) Flower bud collection: take typical tobacco plant main inflorescence free of diseases and insect pests, well-grown flower buds in the single-core marginal stage in sunny days, the appearance of which shows that the calyx and corolla are roughly equal in length, and the bud length is 2.1-2.21cm In the meantime, take the buds and quickly store them in an ice box at 4-7°C;

[0029] (2) Disinfection of anthers: carefully peel off the anthers of the flower buds in step 1 with tweezers in the ultra-clean workbench, then wrap the anthers with sterile commercially available stockings and immerse them in 70% alcohol for disinfection for 15 seconds, quickly take them out and immerse them immediately 10% saturated calcium hypochlorite (Ca(ClO) 2 ) solution for 8 minutes, gently shake the beaker from time to time to make it fully sterilized, and finally ...

Embodiment 2

[0036] Embodiment 2, the in vitro liquid culture of Burley tobacco powder

[0037] (1) Flower bud collection: Take typical tobacco plant side inflorescence free of diseases and insect pests, grow well, and are in the single-nucleus marginal stage flower buds on sunny days. The appearance shows that the calyx and corolla are roughly equal in length, and the bud length is between 2.0-2.28cm. In the meantime, take the buds and quickly store them in an ice box at 4-7°C;

[0038] (2), anther disinfection: use tweezers to carefully peel off the anthers of the flower buds in step 1 in the ultra-clean workbench, then wrap the anthers with sterile gauze and immerse them in 75% alcohol for disinfection for 10 seconds, quickly take them out and immediately immerse them in a concentration of 10 % saturated calcium hypochlorite (Ca(ClO) 2 ) solution for 10 minutes, gently shake the beaker from time to time to make it fully sterilized, and finally rinse with sterile water 3 times, 5 minute...

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Abstract

The invention relates to a high-efficiency tobacco pollen in-vitro liquid culture method, which mainly comprises the operating steps of rapidly releasing a great amount of pollens by rolling a sterilized anther under the effect of B5-13 liquid culture in sterile conditions, conducting low-speed centrifugal separation to collect the pollens, statically placing the pollens in NLN-16 liquid culture mediums containing colchicines with concentration being 50-70mg / L at 32DEG C in the dark for reduplication, then statically placing the pollens in NLN-13 liquid culture mediums at 25 DEG C for embryo induction culture in the dark, placing embryos into a 25 DEG C constant-temperature shaking bed for oscillatory culture in the dark for 7-10 days after the embryos can be seen by naked eyes, transferring the embryos which are at a cotyledon period into solid culture mediums for subculture, and after seedlings are trained, transplanting the seedlings in the field in a transplanting season till blooming, bagging and fruiting. The high-efficiency tobacco pollen in-vitro liquid culture method has the characteristics that the concept is novel, the design is ingenious, scientific and practical, the high efficiency and the stability are realized, the yield is great, the tobacco breeding cycle can be greatly shortened, and the like.

Description

technical field [0001] The invention relates to the field of tobacco tissue culture, in particular to an efficient method for in vitro liquid culture of tobacco pollen. Background technique [0002] Tobacco (Nicotiana tabacum L,.) is an annual herbaceous plant belonging to the family Solanaceae (Solanaceae) and the genus Nicotiana (Nicotiana). Tobacco is an excellent recipient of plant transgenes and a model plant of cytogenetics. It is often used to carry out expression and functional verification of exogenous genes. At the same time, it is used as a plant bioreactor in the field of bioengineering to effectively express some effective Proteins with practical value such as human serum albumin, growth regulator substances, vaccines, antibodies, industrial enzymes, biopolymers, etc. In addition, the neurotoxicity of tobacco nicotine can be used to make pesticides, and malic acid, substances such as citric acid. [0003] Tobacco tissue culture was carried out earlier, mainly ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 蔡长春林国平王毅曹景林张俊杰祖秉桥
Owner TOBACCO RES INST OF HUBEI PROVINCE
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