Cultivation method for inducing autotetraploid of paulownia catalpifolia by colchicine

A technology of autotetraploid and Paulownia catalpa, applied in the field of tree biotechnology breeding, to overcome the cumbersome solid-liquid double-layer culture, save time and cost, and facilitate operation

Active Publication Date: 2016-03-30
HENAN AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, due to the differences in the type and quantity of the cell contents in the leaves of Paulownia catapuliae and other Paulownia, there have been no reports on the successful induction of tetraploid Paulownia catapuliae at home and abroad so far.

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0021] A method for cultivating autotetraploids of Paulownia catalpa with colchicine disclosed in the present invention is realized through the following steps: (1) leaf pretreatment; (2) colchicine induction treatment; (3) bud induction; (4) Root induction; (5) Identification of mutant plants of Paulownia catalpa; (6) Transplanting of tetraploid Paulownia catalpa seedlings.

[0022] The specific steps of the cultivation method of the paulownia catalpa tree autotetraploid are as follows:

[0023] (1) Leaf pretreatment Take 2 to 4 pairs of leaves of Paulownia catalpa near the stem tip, cut the leaves into small square pieces of 1.0×1.0cm2, and place them in MS+0.3mg / In the solid medium of LNAA+14mg / LBA, the temperature was 20 ± 2 ℃ in the dark condition of the artificial climate box and treated for 4~8d; the sucrose concentration in the MS solid medium was 20g / L, and the concentration of agar powder was 3.0 g / L;

[0024] (2) Colchicine induction treatment Under aseptic condi...

Embodiment 2

[0032] A kind of breeding method of paulownia catalpa tree autotetraploid disclosed by the invention is realized through the following steps:

[0033] (1) Leaf pretreatment Take 2-4 pairs of leaves of Paulownia catalpa near the stem tip, cut the leaves into small square pieces of 1.0×1.0cm2, and place them in MS+0.3mg / In the medium of LNAA+14mg / LBA, the concentration of sucrose in MS medium is 20g / L, the concentration of agar powder is 3.0g / L, and the temperature is 20°C in an artificial climate box under dark conditions for 6 days;

[0034] (2) Colchicine-induced treatment Place the leaves of Paulownia catapuliae pretreated in step (1) in a liquid medium containing 30 mg / L colchicine and 2% dimethyl sulfoxide under aseptic conditions, Then place the culture bottle containing the liquid medium in a shaking incubator at 100 rpm, and induce treatment for 48 hours at a temperature of 25° C. in the dark;

[0035] (3) Induction of buds Take out the explants of Paulownia catapuliae ...

Embodiment 3

[0041] A kind of cultivation method of colchicine-induced paulownia catalpa tree autotetraploid concrete steps are as follows:

[0042] (1) Leaf pretreatment Take 2 to 4 pairs of leaves of Paulownia catalpa near the stem tip, cut the leaves into small square pieces of 1.0×1.0cm2, and place them in MS+0.3mg / In the solid medium of LNAA+14mg / LBA, the temperature was 22°C and treated under dark conditions in an artificial climate box for 5 days; the sucrose concentration in the MS solid medium was 20g / L, and the agar powder concentration was 3.0g / L;

[0043] (2) Colchicine induction treatment Place the leaves of Paulownia catapuliae after pretreatment in step (1) in a liquid medium containing 15 mg / L colchicine and 3% dimethyl sulfoxide under aseptic conditions, and then Place the culture bottle containing the liquid medium in a 50rpm shaking incubator, and induce treatment for 36 hours at 24°C in the dark;

[0044] (3) Induction of buds Take out the leaves of Paulownia catapuli...

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Abstract

The invention introduces an effective inductive cultivation method for an autotetraploid of paulownia catalpifolia by using leaves of a diplontic paulownia catalpifolia tissue culture seedling. The method comprises: pre-cultivating the leaves of paulownia catalpifolia for 6 days, wherein the concentration of added colchicine is 30 mg / L, the concentration of dimethyl sulfoxide is 2%, and the soaking time is 48 hours; performing shake cultivation, wherein the induction rate of the tetraploid paulownia catalpifolia reaches 14.5%; detecting that the chromosome number of root tip cells of variant plants of paulownia catalpifolia is 80 by virtue of a chromosome counting method, wherein the chromosome number is two times of that of the corresponding diplontic paulownia catalpifolia; and detecting that the unicellular DNA content of the tetraploid leaves of the paulownia catalpifolia are two times of those of the corresponding diplontic paulownia catalpifolia by using a flow cytometer, showing that a tetraploid paulownia catalpifolia plant is obtained. The cultivation method for inducing autotetraploid of paulownia catalpifolia by colchicine disclosed by the invention has the advantages of being wide in material drawing, simple and convenient to operate, low in cost, high in induction rate and the like, and is of certain guiding significance for woody plants, the tetraploid of which is not easily obtained.

Description

technical field [0001] The invention belongs to the field of tree biotechnology breeding, and in particular relates to a method for cultivating autotetraploids of Paulownia catalpa foliage induced by colchicine. Background technique [0002] Paulownia ( Paulownia ) is a plant of the genus Paulownia in the family Scrophulariaceae. It is a deciduous tree with 9 species and 2 varieties. It is distributed in 25 provinces (cities) and autonomous regions in my country. It is the main choice for important fast-growing timber forests, shelter forests and plain greening in China Tree species play an important role in improving the ecological environment and increasing farmers' income. However, there are still problems in Paulownia production, such as large crowns, low stems and serious arbuscular diseases. Due to the lack of Paulownia germplasm resources, lack of genetic resources, and a serious shortage of new Paulownia varieties that meet the requirements of national economic const...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
Inventor 范国强翟晓巧王杨
Owner HENAN AGRICULTURAL UNIVERSITY
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