A method for improving the ARTP mutagenesis rate of potatoes

Abstract

The present invention provides a method for improving the ARTP mutagenesis rate of potatoes, belonging to the technical field of potato mutagenesis breeding. In the present invention, potato seeds are used as mutagenesis materials, and when the bud eyes start to germinate, they are mutagenized by ARTP for 30 minutes. By selecting appropriate mutagenesis materials, adjusting the mutagenesis period and the mutagenesis time of ARTP, the present invention improves the ARTP mutagenesis rate of potatoes, improves the characteristics of potato varieties, and creates new germplasm resources and breeding intermediate materials.

Description

Technical Field

[0001] The present invention belongs to the technical field of potato mutation breeding, and particularly relates to a method for improving the ARTP mutation rate of potatoes. Background Art

[0002] ARTP (Atmospheric and Room Temperature Plasma) is mutagenesis breeding by atmospheric and room temperature plasma, which refers to a plasma jet that can generate a plasma with a high concentration of active particles (including excited helium atoms, oxygen atoms, nitrogen atoms, OH free radicals, etc.) at a temperature between 25 and 40 °C under atmospheric pressure.

[0003] In the field of plant research, it has been found that treating plant seeds and pollen with ARTP can cause physiological and phenotypic changes in plant seedlings. Treating flax seeds with ARTP can promote the germination rate of seeds under NaCl stress conditions, reduce the relative permeability of the plasma membrane, chlorophyll content, carotenoid content, MDA content, and increase POD activity and soluble protein content; treating Orychophragmus violaceus with ARTP reduces the seedling emergence rate, fresh weight of the aboveground and underground parts of the seedlings, etc.

[0004] The existing potato germplasm resources in China have a narrow genetic background and a short genetic distance, making it difficult to breed breakthrough new varieties. At present, a large number of ARTP breeding studies have been carried out on economic crops such as alfalfa, Gleditsia sinensis, and edible fungi in the world, but there has been no report on potato plasma mutagenesis breeding so far. Summary of the Invention

[0005] In view of this, the purpose of the present invention is to provide a method for improving the ARTP mutation rate of potatoes, improve the potato mutation rate, improve the variety characteristics, and create new germplasm resources and breeding intermediate materials.

[0006] In order to achieve the above invention purpose, the present invention provides the following technical solutions:

[0007] A method for improving the ARTP mutation rate of potatoes, using potato seeds as mutagenesis materials, and when the bud eyes start to germinate, performing ARTP mutagenesis for 30 minutes.

[0008] Preferably, the working gas for the ARTP mutagenesis is He, and the release amount is 12 - 15 SLM.

[0009] More preferably, the rated power of the ARTP mutagenesis is 350 - 400 W, and the distance between the plasma emission source and the mutagenesis material is 8 - 10 cm.

[0010] Preferably, the diameter of the potato seeds is 0.4 - 0.6 cm, and the weight is 0.5 - 1 g.

[0011] Preferably, the potato seed tubers are tissue culture seed tubers.

[0012] Preferably, after collecting the potato seed tubers, they are placed in an environment with a temperature of 22 - 25°C and a light intensity of 1000 - 1500 LX for cultivation until the bud eyes germinate.

[0013] Preferably, it also includes sowing the seed tubers after the mutagenesis is completed. The sowing substrate is a mixture of peat and perlite in a volume ratio of 1 - 2:1 - 2.

[0014] More preferably, it also includes investigating the traits of the mutagenized materials during the flowering period of the potato, including stem color, leaf color, and flower color; and investigating the tuber traits of the mutagenized materials after harvesting, including shape, skin color, and flesh color, to obtain new trait mutants.

[0015] Compared with the prior art, the present invention has the following beneficial effects:

[0016] The present invention provides a method for improving the ARTP mutagenesis rate of potatoes, which provides great assistance for solving the "stuck - neck" problems in the seed industry such as creating new potato germplasm resources and broadening the genetic background of potato germplasm resources.

[0017] The present invention uses potato seed tubers as mutagenized materials. Compared with commercial potatoes, it is convenient for irradiation treatment and has a good mutagenesis effect. Mutagenesis treatment is carried out when the potato bud eyes germinate. At this time, the growth points of the potato plants begin to develop, and it is at the beginning stage of the vigorous growth of the potato, so the mutagenesis effect is better and the mutagenesis rate is high. The mutagenesis time of ARTP is an important factor affecting the mutagenesis rate of potato seed tubers. Controlling the ARTP irradiation treatment for 30 minutes results in a higher mutagenesis rate. Specific Embodiments

[0018] The present invention provides a method for improving the ARTP mutagenesis rate of potatoes. Using potato seed tubers as mutagenized materials, when the bud eyes germinate, ARTP mutagenesis is carried out for 30 minutes.

[0019] The present invention uses potato seed tubers as mutagenized materials, avoiding the problems of large commercial potato tubers, difficult mutagenesis, and poor mutagenesis effect. Mutagenesis is carried out when the potato seed tuber bud eyes germinate. Mutagenesis before germination will not produce a mutagenesis effect; if the mutagenesis time is slightly later, the growth points are too large and are easily damaged during mutagenesis, and they cannot grow and develop normally after sowing. When the bud eyes germinate, the growth points of the potato plants begin to develop, and it is at the beginning stage of the vigorous growth of the potato. At this time, the mutagenesis effect is better and the mutagenesis rate is high.

[0020] The preferred working gas for ARTP mutagenesis of the present invention is He, and the release amount is 12-15SLM (indicating standard state: that is, 1 atmosphere, 1 liter per minute at 25°C, i.e. 1L / min), further preferably 13-15SLM, and more preferably 15SLM; the preferred rated power of ARTP mutagenesis is 350-400W, further preferably 380-400W, and more preferably 400W; the preferred plasma emission source irradiates the mutagenic material at a distance of 8-10cm, and more preferably 10cm. As an implementable embodiment, ARTP mutagenesis is carried out with a plasma mutagenizer (normal pressure room temperature plasma mutagenesis breeding instrument or ARTP mutagenesis breeding instrument, produced by Wuxi Yuanqing Tianmu Biotechnology Co., Ltd.).

[0021] The present invention preferably has a seed potato diameter of 0.4-0.6 cm, more preferably 0.5 cm, and a weight of 0.5-1 g, more preferably 0.8 g. More preferably, the seed potato is a tissue culture seed potato, which has less virus accumulation and more stable mutagenesis effect than commercial potatoes.

[0022] In the present invention, after the potato seed potatoes are collected, they are placed in an environment with a temperature of 22-25° C., more preferably 23° C., and a light intensity of 1000-1500 LX, more preferably 1250 LX, and cultured until the buds sprout.

[0023] The present invention further comprises sowing seed potatoes after the mutagenesis is completed, and the sowing matrix is ​​a mixture of peat and perlite in a volume ratio of 1-2:1-2, and preferably an equal volume ratio. The present invention has no special requirements for management after sowing, and conventional management methods can be used.

[0024] The invention also includes investigating the traits of the induced material when the potato grows to the flowering stage, including stem color, leaf color and flower color; and investigating the tuber traits of the induced material after harvest, including shape, skin color and flesh color, to obtain a mutant with new traits.

[0025] The technical solutions provided by the present invention are described in detail below in conjunction with the embodiments, but they should not be construed as limiting the protection scope of the present invention.

[0026] In the embodiments of the present invention, unless otherwise specified, the methods used are all conventional methods, and the instruments and materials used are all commercially available.

[0027] Example 1

[0028] ARTP mutagenesis method for 'Qingshu 9'

[0029] Variety characteristics: Plant height 97±10.4 cm. Purple stems, dark green leaves, large, hairy, flat leaf margins, large compound leaves, closely arranged, elliptical top leaflets; light red corolla with yellow-green five-star pattern; petal tips white. Oval tubers, red skin and yellow flesh; shallow buds, smooth skin.

[0030] Mutation process:

[0031] (1) Plant tissue culture seedlings of potato 'Qingshu 9' in a seedling tray, manage them under normal growth conditions, and harvest potato seeds, ensuring that the size of the seeds is 0.5 cm in diameter and the weight is 0.5 - 1 g;

[0032] (2) Place the harvested seeds under normal temperature (22 - 25 °C) and low light (1000 - 1500 LX) conditions. Wait until the seeds show germinated eyes and start ARTP plasma mutagenesis;

[0033] (3) Place the germinated seeds (100 seeds, with 10 as blank controls and 30 as mutagenized materials respectively) on the carrier tray, adjust the power to 400 W, the time to 20, 30, 40 min, and the helium release rate to 15 SLM, and start mutagenic breeding;

[0034] (4) Take out the mutagenized potato tubers and sow them in nutrient pots with a specification of 27 cm × 27 cm. The preparation ratio of the cultivation substrate is peat:perlite = 1:1. Sow 1 mutagenized tuber in each nutrient pot, and manage the production after sowing in the same way as normal potato production and cultivation;

[0035] (5) When the potato enters the flowering stage, investigate the differences between the mutagenized plants and the controls. The investigation indicators include stem color, leaf color, and flower color; Hang tags on the mutants with obvious differences and make good test records. After harvest, compare the tuber traits for the final difference evaluation;

[0036] (6) After harvesting the potatoes, investigate the tuber shape, tuber skin color, and flesh color.

[0037] Mutation results:

[0038] Two mutants were obtained when 'Qingshu 9' was treated for 20 min, four mutants were obtained when treated for 30 min, and three mutants were obtained when treated for 40 min. The traits of the mutagenized materials are shown in Table 1. It can be seen that when using the ARTP plasma mutagenesis method for potato mutagenic breeding, the mutagenesis rate is the highest at 13.33% when the irradiation treatment is 30 min.

[0039] Table 1 ARTP mutagenesis results of 'Qingshu 9'

[0040]

[0041]

[0042] Example 2

[0043] ARTP mutagenesis method for 'Chunshu 10'

[0044] Variety characteristics: The plant height is 57 ± 3.7 cm. The stem is green and brown at the base, the leaves are green, the leaf margins are flat, the compound leaves are small, arranged relatively loosely, and the top small leaf is oval; the corolla is light purple. The potato tubers are oval, with yellow skin and yellow flesh; the eyes are relatively shallow and the epidermis is smooth.

[0045] The mutagenesis process is the same as that in Example 1.

[0046] Mutagenesis results:

[0047] Two mutants were obtained when 'Chunshu No. 10' was treated for 20 min, four mutants were obtained when treated for 30 min, and two mutants were obtained when treated for 40 min. The traits of the mutagenized materials are shown in Table 2. It can be seen that when using the ARTP plasma mutagenesis method for potato mutagenesis breeding, the mutagenesis rate is the highest at 30 min of irradiation treatment, reaching 13.33%.

[0048] Table 2 ARTP mutagenesis results of 'Chunshu No. 10'

[0049]

[0050] Example 3

[0051] Using the method of Example 1, ARTP mutagenesis tests were carried out on 'Chunshu No. 3', 'Chunshu No. 11', and 'Chunshu No. 12' respectively.

[0052] The mutagenesis results are shown in Tables 3 - 5. It can be seen that when using the ARTP plasma mutagenesis method for potato mutagenesis breeding, the mutagenesis rate is the highest at 30 min of irradiation treatment.

[0053] Table 3 ARTP mutagenesis results of 'Chunshu No. 3'

[0054]

[0055] Table 4 ARTP mutagenesis results of 'Chunshu No. 11'

[0056]

[0057] Table 5 ARTP mutagenesis results of 'Chunshu No. 12'

[0058]

[0059]

[0060] The above are only the preferred embodiments of the present invention. It should be pointed out that for those of ordinary skill in the art, without departing from the principle of the present invention, several improvements and modifications can be made, and these improvements and modifications should also be regarded as the protection scope of the present invention.

Claims

1. A method for improving the ARTP mutagenesis rate of potatoes, characterized in that, Using potato seeds as mutagenic materials, when the bud eyes start to germinate, they are mutagenized by ARTP for 30 minutes; The rated power of the ARTP mutagenesis is 350 - 400 W, and the distance between the plasma emission source and the mutagenic material is 8 - 10 cm; the working gas for ARTP mutagenesis is He, and the release amount is 12 - 15 SLM; The potato seeds are tissue culture seed potatoes with a diameter of 0.4 - 0.6 cm and a weight of 0.5 - 1 g; It also includes investigating the traits of the mutagenic materials during the flowering period of the potato, including stem color, leaf color, and flower color; after harvesting, investigating the tuber traits of the mutagenic materials, including shape, skin color, and flesh color, to obtain new trait mutants.

2. The ARTP mutagenesis method for high mutagenesis of potatoes according to claim 1, characterized in that, After collecting the potato seeds, they are cultured in an environment with a temperature of 22 - 25 °C and a light intensity of 1000 - 1500 LX until the bud eyes germinate.

3. The ARTP mutagenesis method for high mutagenesis of potatoes according to claim 1, characterized in that, It also includes sowing the seeds after mutagenesis, and the sowing substrate is a mixture of peat and perlite in a volume ratio of 1 - 2:1 - 2.

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