CN111011216B

CN111011216B - Efficient regeneration culture medium and culture method for atlantic potatoes

Info

Publication number: CN111011216B
Application number: CN201911422221.7A
Authority: CN
Inventors: 叶广继; 郑贞贞; 王舰; 周云; 张凤军
Original assignee: Qinghai Academy of Agricultural and Forestry Sciences
Current assignee: Qinghai Academy of Agricultural and Forestry Sciences
Priority date: 2019-12-31
Filing date: 2019-12-31
Publication date: 2023-04-28
Anticipated expiration: 2039-12-31
Also published as: CN111011216A

Abstract

The invention discloses a culture medium for promoting efficient regeneration of atlantic potatoes, and belongs to the field of plant tissue culture. The culture medium consists of a basal culture medium and plant hormone, wherein the plant hormone in the culture medium consists of NAA and ZT, the NAA concentration is 0.01+/-0.005 mg/L, and the ZT concentration is 2.5+/-0.5 mg/L. The culture medium of the invention is used for regenerating the Atlantic potato stem, has simple steps, good effect and great application value.

Description

Efficient regeneration culture medium and culture method for atlantic potatoes

Technical Field

The invention belongs to the technical field of plant tissue culture, and particularly relates to a regeneration culture medium and a culture method of an atlantic potato (called an atlantic potato for short).

Background

Potatoes are plants of the genus Solanum of the family Solanaceae, originating in Andies mountain in south America, cultivated by Indian domestication a minute before ten thousand years, and transferred into China in the open-air ten thousand years. Potatoes are one of four large grain crops in the human society nowadays, are only inferior to rice, corn and wheat, are widely cultivated in China, and are important crops in western regions.

The vegetative tubers are commonly used for asexual reproduction (regeneration) by people, and the material obtaining range is limited. With the development of tissue culture technology, one can use the stem and leaf of potato as explant, add proper amount of plant hormone to the basal medium such as MS, etc. to realize potato regeneration. Regeneration of potatoes comprises two steps of callus induction and seedling differentiation, and the two steps use different hormone proportions.

Atlantic potatoes are a potato variety bred in the United states, introduced into China by the Ministry of agriculture and the national academy of agricultural sciences in 1978, and are the most adopted chip varieties in China at present. Atlantic potatoes are a mainstream variety, and regeneration systems thereof are more reported.

Zhang Zhi the leaf callus induction rate of Atlantic potatoes can be easily up to 100% by using different 6-BA, GA3, NAA and ZT combinations (the establishment of leaf regeneration systems of different varieties of potatoes, the university of Sichuan agriculture, 3 months 2011), but the adventitious bud differentiation rate is lower, and the highest (2.0 mg/L6-BA+0.2 mg/L NAA+1.0mg/L ZT) is only 18.18%.

Liu Rong et al (research on genetic stability of potato and regenerated plants, hunan agricultural university, 2007) found that the hormone ratio of the optimal callus induction medium for atlantic potato was: 2.5mg/L6-BA+0.6 mg/L2, 4-D; according to the orthogonal visual analysis, the three factors NAA, 6-BA and GA3 have the effect of improving the callus differentiation rate of the stem segments of the atlantic strain, and the effect of increasing the average bud number induced by the stem segments and the leaf callus are sequentially increased, and the hormone ratio of the optimal bud differentiation culture medium is as follows: 5.0 mg/L6-BA+0.1 mg/L NAA+10.0mg/L GA3, corresponding to a differentiation rate of 60% and a bud picking day of 40 days. The study also found that the number of days of callus culture was related to the bud differentiation rate, and when the stem-derived callus was used for bud differentiation culture after 21 days of culture (21 days after growing the callus, not 21 days after inoculating the explant), the bud differentiation rate was 73.3% at 35 days.

Disclosure of Invention

The invention aims to provide a novel efficient regeneration culture medium and a culture method of Atlantic potatoes.

The technical scheme of the invention comprises the following steps:

a culture medium for promoting efficient regeneration of Atlantic potatoes comprises a basal culture medium and plant hormone, wherein the plant hormone in the culture medium consists of NAA and ZT, the NAA concentration is 0.01+/-0.005 mg/L, and the ZT concentration is 2.5+/-0.5 mg/L;

by basal medium is meant any medium that can provide sufficient nutrients and suitable pH for potato regeneration in the art.

The culture medium as described above, wherein the basal medium is an MS culture medium containing 2.5% -3.5% (g/mL) sucrose.

As in the previous culture medium, the sucrose content was 3% (g/mL).

The culture medium, wherein the basal medium contains 0.5% -0.7% (g/L) agar.

As in the previous culture medium, the agar content was 0.5% (g/mL).

As in the previous culture medium, NAA concentration was 0.01mg/L and ZT concentration was 2.5mg/L.

A method for promoting efficient regeneration of Atlantic potatoes by culturing the stem segments using the aforementioned medium.

The method as described above, wherein the culturing conditions comprise:

the illumination time is 14+/-2 h/day, and the temperature is 25+/-3 ℃.

Further, the illumination time is 14 h/day, and the temperature is 25 ℃.

According to the method, 2 different culture mediums are not required to be prepared separately for callus induction and bud differentiation, and the stem sections of the Atlantic potatoes can be largely sprouted by using only one culture medium, so that the effect of the 2 culture mediums matched in the prior art is achieved.

In addition, the method of the invention has short time consumption, and the emergence rate can reach 129% only in 43 days; and the prior art generally requires more than 50 days to use.

It should be apparent that, in light of the foregoing, various modifications, substitutions and alterations can be made herein without departing from the spirit and scope of the invention as defined by the appended claims.

The above-described aspects of the present invention will be described in further detail below with reference to specific embodiments in the form of examples. It should not be understood that the scope of the above subject matter of the present invention is limited to the following examples only. All techniques implemented based on the above description of the invention are within the scope of the invention.

Drawings

Fig. 1: emergence of the atlantic potato stem in the medium.

Detailed Description

Example 1A method for regenerating and culturing Atlantic potatoes

1. Materials and methods

1.1 materials

The aseptic test tube plantlet of Atlantic potato growing for about 30d is adopted, the propagation is carried out in an MS basic culture medium, the culture temperature is 25 ℃, the illumination is 14h/d, and the light supplementing is adopted by a fluorescent lamp.

1.2 Medium

NAA was added at a final concentration of 0.01mg/L and ZT was 2.5mg/L based on MS medium containing 3% (g/mL) sucrose.

The formulation (1L) of the basal medium is:

MS medium dry powder (Phytotechnology, M519) 4.43 g+sucrose 30 g+agar 5g, pH 6.0.

1.3 method

Taking stem segments as explants, and spreading 69 total explants on the culture medium of the invention, wherein the length of the explants is 0.4-0.6 cm; the culture temperature is 25 ℃, the illumination is 14h/d, and the fluorescent lamp is adopted for light supplementing.

2. Results

As shown in Table 1, the number of seedlings (adventitious buds) can reach 89 after 43 days of inoculation, and the rate of seedlings is 129%; the total number of the seedling emergence stems is 50, and the seedling emergence stems account for 72.5 percent of the inoculated stems.

TABLE 1 Atlantic stem emergence count

Note that: emergence rate = number of seedlings +.total number of explants x 100%.

The method can lead the atlantic potato to complete two stages of callus induction and seedling emergence by using only 1 culture medium, and can grow a large number of adventitious buds in a short time. The method has very good prospect in industrialized application of Atlantic potato regeneration.

Claims

1. A method for promoting the regeneration of atlantic potatoes, comprising: culturing the stem segments by using a culture medium; the culture medium consists of a basal culture medium and plant hormone, wherein the plant hormone in the culture medium consists of NAA and ZT, the NAA concentration is 0.01mg/L, and the ZT concentration is 2.5 mg/L; the basal medium is an MS medium containing 2.5% -3.5% of sucrose.

2. The method of claim 1, wherein: the sucrose content was 3%.

3. The method of claim 1, wherein: the basal medium contains 0.5% -0.7% of agar.

4. A method as claimed in claim 3, wherein: the agar content was 0.5%.

5. The method of claim 1, wherein: the culture conditions included:

the illumination time is 14+/-2 h/day, and the temperature is 25+/-3 ℃.

6. The method of claim 5, wherein: the culture conditions included:

the illumination time is 14 h/day, and the temperature is 25 ℃.