Method for increasing embryoid emergence quantity in brassica chinensis isolated microspore culture

Abstract

The invention provides a method for increasing embryoid emergence quantity in brassica chinensis isolated microspore culture. The method includes: step one, selecting a brassica chinensis bud in a late mononuclear stage, disinfecting, separating and purifying to obtain isolated microspores, suspending the purified microspores in an NLN liquid culture medium, and adjusting the microspore density to1*10<5>-2*10<5> microspores / mL by a blood counting chamber to obtain microspore suspension; step two, adding the prepared brassica chinensis microspores into a NLN-13 culture medium in which macroelements are reduced by half and 6-BA (0.05mg / L), NAA (0.10mg / L) and activated carbon (100mg / L) are added, subjecting sub-packaged microspore petri dishes to 32 DEG C heat shock treatment in a constant-temperature incubator for 24h, and then transferring into a 25 DEG C constant-temperature incubator for continuing dark culture until embryoids appear. By adoption of the method, an evident promoting effect on brassica chinensis microspore induced embryoid emergence is achieved, the microspore embryoid emergence quantity can be increased. The method has advantages of practicality, specific steps, high repeatability and the like, and extensive application of brassica chinensis isolated microspore culture to haploid breeding can be realized.

Description

technical field [0001] The invention relates to a technique for increasing the number of embryos produced in Chinese cabbage, in particular to a method for increasing the number of embryos produced by culturing free microspores of Chinese cabbage. Background technique [0002] Cabbage heart (Brassica chinesis var.), also known as cabbage sprouts, is a herbaceous plant of the genus Brassica in the genus Brassicaceae. It is domesticated from Chinese cabbage subspecies and Chinese cabbage subspecies. A specialty of South China. The growth cycle of Chinese cabbage is short, and it plays an important role in the annual supply of vegetables, especially the supply of off-season vegetables. Breeding new varieties with high quality and high yield is one of the important goals of Chinese cabbage breeding. [0003] Chinese cabbage is a cross-pollinated plant with obvious heterosis, but homozygosity is very difficult. It usually takes 6-8 generations (years) to cultivate excellent inb...

AI Technical Summary

Problems solved by technology

Although the current technology of microspore culture has made great progress, in practical application, due to the many influences involved in this technology, the change of each factor may cause problems such as poor repeatability and low number of embryos in microspore culture. , the optimum conditions for many factors are for further study

Although researchers have established the free microspore culture system of Chinese cabbage, mustard greens, pepper and other crops, the induction of free microspore embryos among different species has its own rules, which cannot be fully applied. In order to solve the above technical problems, special Propose a new technical solution

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