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Method for increasing non-heading Chinese cabbage microspore embryo induction rate

A technology for head cabbage and embryo induction, which is applied in horticultural methods, botanical equipment and methods, horticulture, etc., and can solve the problems of time-consuming, labor-intensive, inaccurate, etc.

Inactive Publication Date: 2019-06-28
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

These methods are time consuming and inaccurate

Method used

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  • Method for increasing non-heading Chinese cabbage microspore embryo induction rate
  • Method for increasing non-heading Chinese cabbage microspore embryo induction rate
  • Method for increasing non-heading Chinese cabbage microspore embryo induction rate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] 1. Selection of flower buds: Collect samples from 9:00 to 11:00 in the morning, select flower buds (about 2-3mm) with a ratio of petals to anthers close to 1:1, add a small amount of distilled water and store in a 4°C refrigerator.

[0042] 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 IKA control type 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 1 / 2NLN-13, shake and shake well, then centrifuge, pour off the supernatant, leaving a yellow-green precipitate . Then add a small amou...

Embodiment 2

[0045] Embodiment 2: the cultivation of microspores of non-heading Chinese cabbage of different varieties

[0046] Select 14 different varieties (Kaicai, Wuyueman, NHCC-224, Suzhouqing×Aijiaohuang, NHCC-074, NHCC-072, Dafengcai, NHCC-003, NHCC-068, NHCC-088, Erqing . Among them, 'Kaicai', 'NHCC-224', 'NHCC-074', 'Dafengcai', 'NHCC-068' and 'Erqing' are easy-to-embryo varieties, 'Rugao Heitacai', 'Mayuecai' Manman', 'Suzhouqing×Aijiaohuang' are varieties that can produce embryos, and 'NHCC-072', 'NHCC-003', 'NHCC-088', 'NHCC-073', 'NHCC-063' are not easy to produce embryos Variety. Such as figure 1 , such as comparing the results of embryo emergence, among the 14 selected varieties, the embryo emergence rate reached 100%, among which 5 varieties that were not easy to emerge embryos all emerged embryos, and the varieties 'Kaicai' and 'NHCC-224' that were easy to emerge embryos were respectively 26.8 embryos / ml and 12.9 embryos / ml were achieved.

Embodiment 3

[0047] Example 3: Effects of 6-BA and ascorbic acid on the microspore germination rate of non-heading Chinese cabbage

[0048] Selected 'NHCC-224' with a high embryo emergence rate, and carried out research on the microspore embryo emergence rate of 6-BA and ascorbic acid, as shown in Table 1, divided into four cases, control, only 6-BA added, only ascorbic acid added , and adding 6-BA and ascorbic acid at the same time, the comparison results can be found that when nothing is added, the embryo emergence rate of the two varieties is very low, after adding 6-BA and 20mg / L or 30mg / L of ascorbic acid respectively, All of them have different degrees of improvement in the embryo emergence rate, and adding more than 30mg / L ascorbic acid will inhibit the embryo emergence; while adding 6-BA and 20mg / L or 30mg / L ascorbic acid at the same time, the embryo emergence rate will increase again, while Adding 0.08mg / L6-BA and 20mg / L ascorbic acid was the highest rate of embryo emergence, near...

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Abstract

The invention discloses a method for increasing non-heading Chinese cabbage microspore embryo induction rate. The method includes the steps: (1) flower bud collection; (2) microspore embryo culture: sterilizing, cleaning and rinsing the flower buds collected in the step (1), grinding the rinsed flower buds, filtering and centrifuging the grinded flower buds, diluting yellow-green precipitation bya first culture medium, performing heat shock treatment, and performing dark culture for 14-16 days at the temperature of 24-26 DEG C to form an embryoid; (3) enabling the embryoid to sprout to form aplant. The first culture medium is an NLN-13 culture medium added 0.04-0.08mg / L 6-BA, 10-20mg / L ascorbic acid and 1mg / L activated carbon. According to the method, operation steps are simplified whengerm extraction rate can be ensured, operation time is greatly shortened, and the production efficiency of plants is improved.

Description

technical field [0001] The invention belongs to the field of tissue culture, and relates to a method for improving the induction rate of non-heading Chinese cabbage microspore embryos. Background technique [0002] 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 using mi...

Claims

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Application Information

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IPC IPC(8): A01H4/00
Inventor 李英张西林侯喜林王建军刘同坤肖栋
Owner NANJING AGRICULTURAL UNIVERSITY
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