Method for cultivating non-heading Chinese cabbage microspore plants

A technology for headed cabbage and microspores is applied in the field of cultivating non-headed cabbage microspore plants, and can solve the problems of low embryo rate, pollution, cumbersomeness and the like

Inactive Publication Date: 2019-12-10
NANJING AGRICULTURAL UNIVERSITY
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] The present invention mainly aims at improving the problems of the current technology such as cumbersomeness, low germination rate and pollution, thereby providing an efficient and simple method for cultivating non-heading Chinese cabbage microspore plants

Method used

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  • Method for cultivating non-heading Chinese cabbage microspore plants
  • Method for cultivating non-heading Chinese cabbage microspore plants
  • Method for cultivating non-heading Chinese cabbage microspore plants

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0030] 1. Selection of flower buds: Collect samples from 9 am to 11 am, select flower buds with a ratio of petals to anthers close to 1:1, add a small amount of distilled water and store in a refrigerator at 4°C.

[0031] 2. Cultivate microspore embryos: Put the collected samples into a 2ml centrifuge tube, directly add 1% sodium hypochlorite for disinfection for 20 minutes, wash with sterilized water twice for 5 minutes each, and use 1 / 2NLN for the last time -13 for rinsing. Transfer the cleaned sample to the test tube of the sample grinding machine (German LKA control test tube disperser S025), add a small amount of 1 / 2NLN-13, and grind the sample to make the sample fully ground. Prepare a 50mL centrifuge tube, place a 40μm cell filter plug on top of it to filter, centrifuge, pour off the supernatant, then add NLN-13 to shake well, then centrifuge, pour off the supernatant, leaving a yellow-green precipitate. Then add a small amount of NLN-13 to the test tube and dilute to ...

Embodiment 2

[0034] Embodiment 2: the cultivation of microspores of non-heading Chinese cabbage of different varieties

[0035] Choose 7 different varieties (the commercial kind of Jihonghuahua, Jihonghuahua, G112, W1-33, Qianbaolai purple, Qianbaolai green, K2) adopt the method described in embodiment 1 to carry out microspore culture test, and count their Embryonic rate, compare the impact of this invention on the microspore embryonic rate. Among them, 'K2', 'Jihongwa commercial species' and 'G112' are embryo-producible varieties, 'Jihongwa', 'W1-33', 'Qianbaolai purple', 'Qianbaolai green' are not easy Embryo varieties. Such as figure 1 , such as comparing the results of embryo emergence, among the 7 selected varieties, the embryo emergence rate reached 42.8%, of which 3 varieties all emerged embryos, and the other four varieties did not emerge embryos.

Embodiment 3

[0036] Example 3: Effects of different concentrations of EMS on the microspore germination rate of non-heading Chinese cabbage treated at different times

[0037]The 'NHCC-224' with high embryo emergence rate was selected to study the microspore embryo emergence rate of non-heading cabbage treated with different concentrations of EMS at different times, as shown in Table 1, divided into three cases, the control and the same EMS Concentration and different treatment time, different EMS concentration and the same treatment time, by comparing with the control, it can be found that the low concentration of EMS has a certain promotion effect on the embryogenesis of microspores, the concentration is 0.04%, the time is 10 ~ 15min and the concentration is 0.08%. A time of 10 minutes is beneficial to embryogenesis. When the concentration of EMS is 0.08% and the treatment time is 10 minutes, it can obviously promote embryo emergence, which is about doubled. When the concentration of EMS...

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Abstract

The invention discloses a method for cultivating non-heading Chinese cabbage microspore plants. The method comprises the following steps: (1) selecting flower buds; (2) cultivating microspore derivedembryos, wherein the flower buds are sterilized and washed and rinsed with a 1/2NLN-13 culture medium; the rinsed flower buds are added into the 1/2NLN-13 culture medium and fully ground, filtered andcentrifuged, a yellow-green precipitate is diluted with a NLN-13 culture medium and split-charged into a culture dish after treatment with ethylmethane sulfonate, activated carbon is added into the culture dish to enable the concentration of the activated carbon to be 1 mg/mL, heat shock treatment is carried out, and then dark culture is carried out for 14 days to form embryoid bodies; (3) germinating mature embryos into plants, wherein the embryoid bodies undergo shaking culture, the mature embryos becoming green are transferred to an improved solid 1/2MS culture medium for 40 days, and thenacclimatization and transplanting are directly carried out. According to the method, under the condition that the embryo yield can be guaranteed, the operation steps are simplified, the operation time is greatly shortened, and the plant production efficiency is improved.

Description

technical field [0001] The invention belongs to the technical field of plant tissue culture, in particular to a method for cultivating non-heading Chinese cabbage microspore plants. [0002] technical background [0003] Chinese cabbage is mainly a cross-pollinated plant. In traditional breeding work, it is necessary to obtain purified parents only through conventional selfing. Due to selfing decline, the selection of homozygous parents is a very difficult problem, and it takes a long time and consumes manpower. As well as financial resources, the breeding efficiency is very low. Microspore culture technology is the process of cultivating a single cell into a homozygous plant by utilizing the totipotency of the cell, which can effectively shorten the breeding period, obtain a large number of homozygous plants in a short period of time, and greatly improve the breeding efficiency. Lichter first discovered on Brassica napus in 1982 that embryoid bodies could be obtained by usi...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01H4/00
CPCA01H4/001A01H4/008
Inventor 李英汪家礼侯喜林王建军刘同坤张昌伟
Owner NANJING AGRICULTURAL UNIVERSITY
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