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Composite multiplying agent for plant chromosome and multiplying method

A chromosome doubling and chromosome technology, applied in the fields of botanical equipment and methods, plant genetic improvement, application, etc., can solve the problems of low success rate of colchicine doubling, abnormal growth, and large damage to plant materials.

Active Publication Date: 2018-05-04
SHENYANG AGRI UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The chromosome doubling technique often used in the past is to use colchicine as a doubling agent, but the doubling effect of colchicine on plants is difficult to meet the needs of the above research. Abnormalities, deformities and even death of processed material, in addition colchicine has a lower doubling success rate
Moreover, colchicine is highly toxic, and has greater toxicity and potential carcinogenic effects on mammals including humans, and is very harmful to the health of operators and environmental safety.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0029] Example 1 Chromosome doubling of monocot rye

[0030] 1. Reagent preparation

[0031] 1. Preparation of doubling agent mother liquor

[0032] The above-mentioned doubling agents need to be separately formulated into mother liquors, and when used, they should be diluted according to the formula requirements. Each mother liquor is uniformly prepared according to the following method. The preparation method of reagents / medicaments is a conventional technique that is all adopted in ordinary laboratories:

[0033] Measure 50ml of dimethyl sulfoxide into a 1000ml volumetric flask, calculate and weigh the corresponding amount of solid drug according to the effective content of each drug according to the final volume of 1000ml and final concentration of 10mM, and add it to dimethyl sulfoxide, stirring slowly until it is completely dissolved; add 200ml of ethylene glycol, stir well until it dissolves evenly; add distilled water to make up to 1000ml, stir well, put it into a bro...

Embodiment 2

[0044] Example 2 Chromosome doubling of dicotyledon pea

[0045] 1. Reagent preparation

[0046] 1. Preparation of doubling agent mother liquor

[0047] Same as Example 1

[0048] 2. Preparation of compound doubling agent

[0049] Add 500ml of sterilized distilled water into a 1000ml volumetric flask, and use a pipette to absorb the above reagent mother liquor and add them to the distilled water. 3 ml, 1 ml of fenpyramide, 1 ml of dithiazide, and 1 ml of dithiopyr, and then make up to 1000 ml with distilled water. The final concentrations of the above components in the solution were 50 μM of secbutaline, 30 μM of phosphinothricin, 30 μM of amifluralin, 10 μM of pentoxynamide, 10 μM of fenthiazide, and 10 μM of dithiopyr.

[0050] 2. Preparation and pretreatment of plant material to be doubled - seed treatment and germination

[0051] Take pea seeds with germination ability, soak them overnight (14-18 hours) in water at room temperature (about 18-25°C) to make the seeds sw...

Embodiment 3

[0062] Example 3 Chromosomal doubling of maize haploid immature embryos

[0063] 1. Reagent preparation

[0064] 1. Reagent mother solution preparation

[0065] Same as Example 1;

[0066] 2. Preparation of compound doubling agent

[0067] Add 60ml of sterilized distilled water into a 100ml volumetric flask, and use a pipette to absorb the above reagent mother liquor and add them to the distilled water. ml, 0.3 ml of chlorpropen, 0.1 ml of pendimethalin, 0.1 ml of pendimethalin, and 0.1 ml of metolachlor, then dilute to 100 ml with distilled water. The final concentrations of the above components in the solution are 50 μM of glufosinate-methyl, 50 μM of nocodazole, 30 μM of chlorphenamine, 30 μM of chlorphenamine, 10 μM of pentoxychlor, 10 μM of pendimethalin, and 10 μM of pendimethalin, and Amine 10 μM.

[0068] 3. Preparation of antidote: add 0.2g of potassium dihydrogen phosphate to 100ml of distilled water to prepare a 0.2% aqueous solution, and then add 0.5g of urea;...

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PUM

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Abstract

The invention discloses a reagent formula for multiplying a plant chromosome by using a composite reagent without colchicines and a multiplying method. The plant chromosome is multiplied by using a composite formula prepared from multiple mitosis inhibitors and tubulin inhibitors; compared with a conventional chromosome multiplying method, the method has the characteristics of low toxicity, high efficiency and wide application range; cooperative use of three compounds such as a multiplying agent, an aid and a detoxicant with different functions is the key point of the reagent formula, and functions of improving multiplying effects and reducing medicine damage are achieved; by adopting the reagent formula, the problems that growth abnormity, malformation or even death of a treated plant material are generally caused, and the multiplication success rate is relatively low since the treated plant material is seriously damaged in conventional process that a plant chromosome is multiplied byusing colchicines, are solved; and more importantly, the problems that the colchicines are highly toxic, have potential carcinogeneses for mammals including human beings, and can cause serious harm to health of operators and environment security, can be solved.

Description

technical field [0001] The invention relates to a doubling agent formula and a doubling method for plant chromosomes, belonging to the fields of plant biology, plant biotechnology and plant breeding, in particular to a method for obtaining plant polyploids and doubling haploids to become doubled haploids (also known as haploids) double haploid, pure and diploid) methods. Background technique [0002] Ploidy change in plants is an important research content in plant biology research, plant biotechnology and plant breeding. Compared with diploid, autopolyploid has the characteristics of enlarged organs or increased content of metabolites, and has excellent breeding and utilization value for crops for the purpose of harvesting vegetative organs and asexually propagated crops; artificially created multiple Ploidy can also recombine the genetic material of wild species and cultivated species to breed new crops; haploid doubling can obtain pure lines of genotype homozygous plants...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H1/08
CPCA01H1/08
Inventor 姜海鹰徐玉波宝琴张奇艳周亚圣
Owner SHENYANG AGRI UNIV
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