Dihaploid induction method of head cabbage with high efficiency

A technology of head cabbage and double haploid, applied in horticultural methods, botanical equipment and methods, horticulture, etc., can solve the problems of inability to meet large-scale embryo production and efficient breeding, low transplanting survival rate, and weak rootlets of seedlings and other problems, to achieve the effect of eliminating the problem of vitrification, high embryo emergence rate, and green and strong performance

Inactive Publication Date: 2013-06-05
陕西省杂交油菜研究中心
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  • Abstract
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  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, the microspore culture technology of head cabbage in my country is not yet mature enough, and most of them are in the digestion and absorption of related species culture technology and its limited improvement. The amount of embryos is very small; or the process of embryo formation and seedling formation is easy to vitrify, the seedlings have weak roots, and the survival rate of transplanting is low, which is far from meeting the requirements of large-scale embryo production and efficient breeding

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  • Dihaploid induction method of head cabbage with high efficiency
  • Dihaploid induction method of head cabbage with high efficiency
  • Dihaploid induction method of head cabbage with high efficiency

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] The formulations of various media used in this example are shown in Table 1.

[0025] In this example, several cabbage genotypes of Q2, Q4, Q5, Q8, HP63 and HW06 were selected as raw materials, and the double haploids of cabbage were induced and cultivated according to the following steps, and a control group was set up at the same time. In the group, there was no colchicine doubling agent in the priming medium, and no paclobutrazol in the subculture and rooting medium:

[0026] (1) Selection of flower buds and isolation of microspores

[0027] To be grown in the artificial climate box, glass greenhouse and natural environment of the field to the initial flowering stage of cabbage, combined with microscopic examination, the size of flower buds is in the range of 3.5-5.5mm, and it is determined that the proportion of microspore development in the single-nucleus marginal stage reaches 70% For the above, select the corresponding flower buds and put them into the steel bas...

Embodiment 2

[0042] Example 2 In the process of isolation, initiation and embryo induction, the concentration of sucrose in each medium was determined by experiment

[0043] In the mid-April, 2007 flowering period, get 20 parts of head cabbage in Daejeon, the flower bud (bud size is between 3.5-5.5mm) that the proportion of microspores exceeds 70% in the marginal stage, put it into a steel basket for sterilization, and put it in a steel basket. The microspores were crushed and washed in B5 liquid medium with a sucrose concentration of 150g / L to separate the microspores, and the isolated microspores were carried out in NLN liquid medium with a sucrose concentration of 130, 140, 150, 160, 170 and 180g / L at 33°C. After high-temperature cultivation, after 2 days, observe and record the expansion of microspores, as shown in Table 3.

[0044]Table 3 The influence of different sucrose concentrations in the NLN liquid medium on the development and expansion of microspores during the start-up cultu...

Embodiment 3

[0052] Example 3 Experiments on the appropriate concentration and treatment duration of synchronous doubling induction with colchicine in the start-up culture stage

[0053] In April 2007 and 2008, buds were taken from the field at the flowering stage, separated and washed with B5 liquid medium with 150g / L sucrose concentration, NLN liquid medium with 160g / L sucrose concentration was used to start induction, and NLN liquid medium with 140g / L sucrose concentration was used to induce embryo formation , the doubling induction is to add different concentrations of colchicine in the NLN liquid medium at the start-up culture stage, and after a certain period of time (48 hours), investigate the expansion rate and the final embryo doubling situation. The results are shown in Table 5.

[0054] Table 5 Different concentrations of colchicine are treated with the doubling of embryos for 48 hours

[0055] Colchicine

[0056] Experiments have shown that colchicine can double the ...

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Abstract

The invention discloses a dihaploid induction method of head cabbage with high efficiency, which belongs to the filed of biological breeding. The method relates to a regeneration of microspore plant of the horticultural plant head cabbage and an improvement of dihaploid induction technology thereof. The method comprises the following steps: (1) selecting bud and separating microspore; (2) doubling inducing and culturing formed embryo; (3) differencing seedlings and subculturing; (4) cultivating strong seedling rooting. According to the invention, the dihaploid of head cabbage can be applied in a mass production with a large scale, a plurality of novel genetype pure lines can be provided for breeding. The method is affected slightly of the growth environment and genotype, and has the advantages of high embryo generation and embryo formation, good doubling effect, strong growth of seedlings plant, less subculture frequency, strong root and seedings after rooting and high survival rate of the field transplanting.

Description

technical field [0001] The invention relates to a method for inducing double haploid (DH) of plants, in particular to a regeneration method of horticultural crop head cabbage microspore plants and a method for efficiently inducing double haploid, and a culture medium thereof. Background technique [0002] Heading cabbage is a kind of edible vegetable with large cultivation area, wide adaptability and high nutritional value, which is deeply loved by the people. However, because it is native to the Mediterranean coast, domestic germplasm resources are scarce, and foreign varieties and parental resources with superior performance are not available. It is difficult to obtain, which has always restricted the development of the breeding of fine varieties of cabbage in my country. If double haploid breeding is adopted, it will inevitably create a new and effective way for the acquisition of fine germplasm resources and the cultivation of new varieties, and it will also greatly improv...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01H4/00
Inventor 王灏赵小萍李殿荣田建华同晓丽
Owner 陕西省杂交油菜研究中心
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