Method for rapid induction of homogenous polyploids of Chinese jujube

Abstract

The invention relates to a method for rapid induction of homogenous polyploids of Chinese jujube, that is the method can double and further develop chromosomes of a single-cell embryo into homogenous tetraploids by simultaneously carrying out induction with colchicine during the somatic embryogenesis process of in vitro leaf blades of the Chinese jujube. The method comprises the steps of somatic embryo induction, doubling of the chromosomes, development of embryoids till nature and detection and screening of the homogenous tetraploids. The specific steps are as follows: selecting tissue culture seedling leaf blades of the Chinese jujube with robust growth and seedling age of 20-30d, and using a pair of scissors after sterilization to shear 5-8 times by being vertical to nervures; then inoculating the leaf blades in an induction culture medium MS plus 15-25g / L of maltose plus 3.0-5.0g / L of agar plus 5.0-10.0mg / L of TDZ plus 1.0-2.0mg / L of AgNO3 by leading the front surfaces to face upwards, and simultaneously adding 15-20mg / L of the colchicine into the culture medium, and carrying out dark culture for 40-50 days; transferring embryoids differentiated from calli of the leaf blades of the Chinese jujube into a hormone-free culture medium of the MS plus 40g / L of sucrose plus 500mg / L of caseinhydrolysate for lighting culture and further developing the embryoids into somatic embryo seedlings; and screening out the homogenous tetraploids from the somatic embryo seedlings through chromosome detection or the detection by a flow cytometer, wherein the pH of the culture medium is 5.0-6.0, and the culture temperature is 25-30 DEG C. The method can avoid the appearance of chimeras, omit the steps of separation and purification, rapidly obtain the homogenous polyploids and have important application value for accelerating the breeding process of the polyploids of the Chinese jujube.

Description

technical field [0001] The invention relates to a method for rapidly obtaining jujube homopolyploid under tissue culture conditions. Background technique [0002] The artificial creation of polyploidy opens up new avenues for breeding efforts. Polyploidy of fruit trees generally has the advantages of vigorous growth, thick branches, thick leaves, large fruit, few or no seeds, high yield, strong adaptability and stress resistance, etc., and these advantages can be maintained through asexual reproduction. Therefore, polyploid breeding, which has outstanding advantages such as short cycle, quick results, and labor-saving, has become the first choice of many breeders. In recent years, the use of colchicine to induce polyploids has developed rapidly. However, due to the use of stem tips as mutagenic materials, most of the obtained polyploids are chimeras, which are prone to back mutations. Selection, purification, and increasing the proportion of polyploid cells to achieve the ...

AI Technical Summary

Problems solved by technology

However, due to problems such as small flowers, difficult emasculation and pollination, and severe embryo abortion, it is very difficult to cross breed jujube trees, making polyploid breeding more important in jujube trees

The colchicine induction of jujube through the organogenesis pathway using young shoot tips has been successful, but the colchicine induction of jujube through the somatic embryonic pathway has not been reported

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