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

A technology for culturing microspores and embryoid bodies, which is applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of incomplete application, poor reproducibility of microspore culture, and low number of embryos, and achieves improved embryo production. number, the effect of shortening the breeding cycle

Inactive Publication Date: 2020-01-31
海南省农业科学院蔬菜研究所
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  • Application Information

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 furth...

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

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Experimental program
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Effect test

Embodiment 1

[0024] Embodiment 1, the influence of basal medium on the induction of cabbage microspore embryoid bodies

[0025] It can be seen from the table below that different media have different effects on the induction of microspore embryoids in Chinese cabbage. In this study, it was found that the basic medium (1 / 2NLN-13) with halved macronutrients was beneficial to the induction of microspore embryos in Chinese cabbage. The number of embryos was significantly higher than that of NLN-13. In addition, the different genotypes have a great influence on the emergence of microspores. Among the 20 materials tested, 8 were induced to produce embryos, and the induction rate reached 40.00%. Among them, the number of embryos from April to September was the highest, followed by 70 days for pointed leaves, other materials are difficult to induce embryos.

[0026]

[0027]

[0028] Among them, NLN-13 is a sucrose concentration of 13%; 1 / 2NLN-13 is an NLN medium in which macronutrients are...

Embodiment 2

[0029] Embodiment two, the influence of different hormone contents on the microspore embryogenesis of cabbage heart;

[0030] refer to figure 1 It can be seen that the ratio of hormone types and concentrations is closely related to the induction of microspore embryoid bodies in Chinese cabbage. When the 6-BA content was 0.05 mg / L, the number of embryoid bodies increased with the increase of NAA content, but when the 6-BA concentration was 0.1 mg / L and 0.15 mg / L, the NAA concentration was too high. Higher levels will inhibit the production of embryoid bodies. In general, 0.05mg / L 6-BA+0.10mg / LNAA is the most suitable hormone combination for embryoid body induction medium for microspore culture in Chinese cabbage.

Embodiment 3

[0031] Embodiment three, add gac on the impact of Chinese cabbage microspore embryogenesis;

[0032] refer to figure 2It can be seen that adding a small amount of activated carbon is beneficial to the induction of microspore embryoid bodies of Chinese cabbage. When the concentration of activated carbon is 100 mg / L, the yield of microspore embryoid bodies of each test material is significantly higher than that of other treatments. The number of embryoid bodies decreases accordingly.

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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...

Claims

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Application Information

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IPC IPC(8): A01H4/00
CPCA01H4/005
Inventor 庞强强周曼蔡兴来孙晓东张文
Owner 海南省农业科学院蔬菜研究所
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