CN110710452A - Tissue culture and rapid propagation method of eriobotrya japonica - Google Patents
Tissue culture and rapid propagation method of eriobotrya japonica Download PDFInfo
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- CN110710452A CN110710452A CN201911071911.2A CN201911071911A CN110710452A CN 110710452 A CN110710452 A CN 110710452A CN 201911071911 A CN201911071911 A CN 201911071911A CN 110710452 A CN110710452 A CN 110710452A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
Abstract
The invention discloses a tissue culture and rapid propagation method of eriobotrya japonica seedlings. According to the invention, young loquat fruits are collected, the surfaces of the loquat fruits are disinfected, young loquat embryos are stripped, callus is induced, and then the young loquat embryos are differentiated on a culture medium to induce and regenerate plants, so that the technical problem of difficulty in primary generation of loquat is solved, the propagation coefficient of loquat tissue culture seedlings is greatly improved, a large number of aseptic loquat tissue culture seedlings can be obtained in a short time, and materials are provided for fine loquat seed breeding and genetic improvement of genetic engineering.
Description
Technical Field
The invention belongs to the field of tissue culture, and relates to a method for tissue culture and rapid propagation of seedlings of eriobotrya japonica.
Background
Eriobotrya japonica belongs to Rosaceae, and belongs to the genus of fruit tree. The loquat fruit is sweet in taste, is a health-care fruit with rich nutrition, and also has the medicinal values of promoting the production of body fluid to quench thirst, treating lung heat cough and asthma and the like. In the conventional breeding of the loquat, the difficulty is higher due to the reasons of long childhood period, high gene heterozygosity and the like. On the other hand, in the loquat seed propagation, due to high heterozygosis of genes, the segregation of seedling offspring is larger. With the development of biotechnology, genetic engineering means are used for improving crops, and the important role is played more and more. While tissue culture is the precursor for genetic improvement of crops by genetic engineering. The tissue culture of the loquat can not only accelerate the breeding of improved varieties, but also provide materials for the modern genetic engineering breeding. The tissue culture of loquat is difficult to obtain aseptic tissue culture seedlings by common methods such as stem tip disinfection, mainly because the young leaves of loquat contain a large amount of fuzz and are difficult to obtain the primary aseptic tissue culture seedlings by the disinfection of conventional explants (stem tips); on the other hand, the stem tip of loquat and the like have high content of phenolic substances, so that browning phenomenon easily occurs in the culture process, and the tissue culture seedling is dead.
Disclosure of Invention
The invention aims to provide a method for tissue culture and rapid propagation of seedlings of eriobotrya japonica.
The purpose of the invention can be realized by the following technical scheme:
a method for tissue culture and rapid propagation of seedlings of Eriobotrya japonica comprises the following steps: picking young loquat fruits with the transverse diameter of 1.0-1.5 cm from the middle and the last ten days of 3 months to 4 months in Jiangsu area, pretreating at the low temperature of 4 ℃ for 12-15 days, removing sepals at the bottoms of the fruits, removing fuzz on the surfaces of the fruits, disinfecting and cleaning, stripping young loquat fruits on a clean bench, inoculating the young loquat fruits on an MS basic culture medium with 2mg/L6-BA and 0.5 mg/L2, 4-D callus induction culture medium, and carrying out dark culture at the temperature of 23-25 ℃ for 45-50 days to induce callus formation; subculturing the callus for 1 time until the callus grows to 1cm2Inoculating to a bud regeneration culture medium for culturing for 55-60 days to induce the formation of buds, wherein the bud regeneration culture medium is an MS culture medium added with 2mg/L ZT +0.2mg/L IAA; when two leaflets grow out from the buds of the callus, the cut buds are transplanted to an MS culture medium containing 2mg/L ZT for subculture for 55-65 days, and the newly cut tissue culture seedlings after the subculture are inoculated into a rooting culture medium of 1/2MS basal medium added with 0.4mg/L IBA for culture for 30-35 days to obtain the white loquat seedlings.
The culture temperature of each step of the method is 23-25 ℃, except that the callus needs to be induced in dark and dark, the illumination intensity of other culture steps is 3000-5000 Lx.
The method is preferably carried out in Jiangsu area for 3 months and 20-25 days or 4 months and 8-10 days, the transverse diameter of young loquat fruits is 1.0-1.5 cm, low-temperature pretreatment is carried out for 15 days at 4 ℃, sepals at the bottom of the fruits and downy feathers on the surfaces of the fruits are removed, the fruits are washed for four hours with running water, the surfaces of the fruits are disinfected with 70% alcohol for 10min, then disinfected with 30% sodium hypochlorite for 15min, and washed with sterile water for 3-4 times. And stripping the young embryo of the loquat young fruit on a clean bench.
The method preferably pre-treats the young loquat fruits at 4 ℃ for 15 days.
The method preferably comprises the step of inoculating the immature embryo to a callus induction culture medium, and performing dark culture at 23 ℃ for 50 days to induce callus formation.
Has the advantages that:
according to the invention, young loquat fruits are collected, the surfaces of the loquat fruits are disinfected, young embryos of the loquats are stripped, callus is induced, and then the young loquat fruits are differentiated on the culture medium to induce and regenerate plants, so that the technical problem of difficulty in primary generation of the loquats is solved, the propagation coefficient of loquat tissue culture seedlings is greatly improved, a large number of aseptic loquat tissue culture seedlings can be obtained within about 6 months by optimizing the culture conditions and the culture mediums, and materials are provided for fine loquat seed breeding and genetic improvement of genetic engineering.
Drawings
FIG. 1 shows that the loquat young fruit has 5 different sampling periods Bar 2cm
Note: i, 3 months and 9 days; II: 3, month and 17 days; III: 24 days after 3 months; IV: 3, month and 30 days; v: 4 month and 10 days
FIG. 2 proliferation of shoots
Note: a: multiplication of loquat sprouts; b: growth of shoots after 60 days.
FIG. 3 tissue rapid propagation process of Eriobotrya japonica
Note: a: callus induction; b, C, D, inducing the bud for 30 days; e, F, G, inducing the bud for 90 days; h: subculturing; i: rooting culture
Detailed Description
1 materials and methods
1.1 test materials
The test was started in 2018, 3 months, and the test sample was taken from the loquat resource garden in Suzhou Taihu evergreen fruit tree technical promotion center. The test materials are high-quality Baiyu and Guanyu which are popularized and planted in Suzhou areas, and the test trees have intermediate vigor and good flowering and fruit setting. And selecting fruits with basically consistent tree vigor and small flowering time difference for picking. After sampling, the samples were rapidly processed by cold treatment at the university of agriculture laboratory, Nanjing, brought back, Suzhou.
1.2 design of the experiment
Orthogonal tests are adopted in the experimental design to study the influence of different development periods, 2,4-D concentration and low-temperature treatment time of the young embryos on the callus induction rate of the young loquat embryos.
The test takes five times of sampling, wherein the five times of sampling comprise A:3 months and 9 days, B:3 months and 17 days, C:3 months and 24 days, D:3 months and 30 days, and E:4 months and 10 days. 120 fruits of 'white jade' and 'crown jade' are respectively sampled each time, and relatively full and healthy fruits are selected, the sizes of the selected fruits are consistent to the greatest extent, the time for flowering and fruit setting is ensured, and the development period of the sampled fruits is ensured to be not greatly different.
The low-temperature pretreatment is designed to be at three levels of 0d, 7d and 15d, and the low-temperature pretreatment temperature is 4 ℃.
Three 2,4-D concentrations were set up for the experiment, as shown in Table 1.
TABLE 1 combination of growth regulators for immature embryo callus induction
Using an orthogonal design with IBM SPSS Statistics 25 software, the system gave 25 processing groups with the results shown in table 2.
TABLE 2 orthogonal analysis table for callus induction test of immature embryo
1.3 test methods and assays
1.3.1 explant treatment
Removing all sepals at the bottom of the fruit, scraping off the surface of the peel and the villi in the sepals with an operating knife, washing with running water for four hours, draining off water, placing into a super-clean workbench, soaking in 70% alcohol for 10min, taking out, washing with sterile water twice, placing in 30% sodium hypochlorite for 15min, taking out, and washing with sterile water for multiple times.
1.3.2 Primary culture of loquat immature embryos
MS culture medium is used as basic culture medium (Beijing Soilebao), agarose 5.5g/L and cane sugar 30g/L are added, pH is regulated to 5.8-6.0, after autoclaving, a suction filtration sterile growth regulator is added, and the mixture ratio of the culture medium and the growth regulator is ① 6-BA (2mg/L) +2,4-D (0mg/L), ② 6-BA (2mg/L) +2,4-D (0.3mg/L), ③ 6-BA (2mg/L) +2,4-D (0.5mg/L) and ④ 6-BA (2mg/L) +2,4-D (0.7 mg/L).
The fruit was dissected with a scalpel, the seeds were removed with forceps and the embryos were removed by transecting a knife at the seed tip 1/3 as described in Tokukukukukukuhiki (1982) et al, the remaining part was cut in two and placed up on the medium, 5-8 explants were placed in each dish, and each treatment was repeated 5 times. Dark culture is carried out at 23 ℃, and the induction rate of the callus is counted after 45 days of culture. Subculturing the callus for 1 time until the callus grows to 1cm2Inoculating to redifferentiation culture medium to induce the formation of bud.
1.3.3 regeneration of shoots
In the bud induction stage, an MS culture medium is still used as a basic culture medium, the proportion of plant growth regulators is set to 7 treatments, each treatment is repeated for 3 times, 5 callus tissues are placed in each repetition, and non-browned and compact callus tissues are selected. After the callus is inoculated to a dedifferentiation culture medium for two months, the browning condition of the callus, the condition that the callus is changed into green callus and the number of seedlings obtained by each culture medium growth regulator formula group are counted, so that a culture medium formula which is most suitable for redifferentiation of the loquat callus is researched.
TABLE 3 bud regeneration Medium growth regulating substance formula
1.3.4 subculture of regenerated shoots
The cut buds are directly placed on an MS culture medium added with 2mg/L ZT for culture, and the quick growth and tillering of the buds are promoted. Subculturing once every 30-45 days along with the growth of the seedlings.
1.3.5 rooting of seedlings
In the experiment, a rooting culture medium 1/2MS is selected as a basic culture medium, IBA (0.4mg/L) is added, and the rooting condition is observed.
1.3.6 analysis software and Instrument
Performing data statistics and arrangement by using an Excel table; data analysis the difference significance analysis was performed using IBM SPSS Statistics 25 software.
Kedi K-2258 hand paraffin slicer, Kedi H-I baking machine, electronic vernier caliper, come card type microscope
2. Results and analysis
2.1 analysis of the major factors affecting callus formation
In order to research the influence of low-temperature pretreatment and 2,4-D concentration on callus formation of loquat endosperm at different sampling time and different duration, orthogonal analysis is carried out on test data, and the result is shown in Table 4, wherein the Sig value shows that the two factors of the sampling period and the low-temperature pretreatment have significant difference in a 95% confidence interval, which shows that the two factors have great influence on callus formation; the 2,4-D concentration did not show significant difference, indicating that the three concentration gradients set up in this experiment did not show a large effect on callus formation, which is essentially consistent in the performance of two different varieties of loquats, white jade and crown jade. From the F value analysis, it can be seen that in the orthogonal analysis of white jade loquat, 20.173 (sampling period) > 3.387 (low temperature pretreatment) > 1.376(2,4-D concentration); the expression in the crown jade is as follows: 13.210 (sampling period) > 8.679 (low temperature pretreatment) > 0.737(2,4-D concentration). The larger the F value in the orthogonal analysis is, the larger the influence of the factor on the index is, so that the main and secondary influence relationship of the factor for obtaining the callus induction rate of the loquat is as follows: sampling period > Low temperature pretreatment > 2,4-D concentration.
To further explore the optimal levels of each factor, and to obtain the optimal treatment combinations for this trial, we performed multiple duncan comparisons for each factor.
TABLE 4 analysis of variance table of orthogonal test results of white sand loquat
Note: significant level of difference 5%.
TABLE 5 Duncan multiple comparison of sampling periods
Note: significant level of difference 5%; a, sampling at 3 months and 9 days, B, sampling at 3 months and 17 days, C: sample on 24 days 3 months, D: sample 3 month and 30 days, E: samples were taken at 4 months and 10 days.
Table 5 shows the average callus formation rates and the differential analysis of the groups at different sampling periods, and it can be seen that the embryo callus formation rate of ` Baiyu ` was highest in the fifth sampling (group E) and was 44.30% in average, and that the endosperm callus formation rate in the third sampling (group C) was second to 30.90%. The embryogenic callus formation rate of 'crown jade' occurred at the highest rate in the third (group C) and fifth sampling period (group E), but there was no significant difference. Therefore, the third sampling (group C) and the fifth sampling (group E) of the white sand loquat 'white jade' and 'crown jade' are ideal sampling periods for inducing callus by embryos, and the callus induction rate is the highest.
TABLE 6 Duncan multiple comparison of Low temperature pretreatment time
Note: significant level of difference 5%.
As shown in Table 6, in the low-temperature pretreatment of different durations, the callus formation rate of the white jade embryos reaches 23.61 percent in 15 days at most in numerical view, which is obviously higher than that of the white jade embryos without the low-temperature pretreatment; but the low-temperature pretreatment for 7 days of treatment does not produce significant difference with the pretreatment-free treatment; the low-temperature pretreatment is carried out for 15 days, the embryo callus induction rate of the corolla reaches 54.24 percent, which is obviously higher than that of the corolla without the low-temperature pretreatment and the low-temperature pretreatment for 7 days. Thus, the low temperature pretreatment was selected to be optimal for 15 days.
TABLE 82 Duncan multiple comparison of 4-D concentrations
Note: significant level of difference 5%.
As can be seen from Table 8, no significant difference was shown between the 2,4-D concentration levels in the mean callosity rates of the white jade and the crown jade. However, the callus obtained from the 0.5 mg/L2, 4-D treated group was denser and more conditioned than the other two 2,4-D level callus.
TABLE 9 statistics of callus formation rate of loquat C-15 group
Note: c-15 represents the third sampling, low-temperature pretreatment for 15 days; significant level of difference 5%.
By combining the analysis, the cross diameter of the fruit is 10 mm-12 mm, the fruit is sampled when the pericarp is green but still has yellow villus (group C), the pretreatment is carried out for 15 days at low temperature, and MS +6-BA (2mg/L) +2,4-D (0.5mg/L) is adopted as the combination of the culture medium formula, so that the embryo induction callus efficiency is highest.
2.2 shoot regeneration
TABLE 10 statistics of callus differentiation of Eriobotrya japonica
Note: significant level of difference 5%
As can be seen from Table 10, after the callus was inoculated into the differentiation medium for two months, white eriobotrya japonica had the highest non-browning rate of the callus on ② -treated medium, which was 46.67%, but had no significant difference between the treatments, treatment ⑤, which was significantly higher than the other treatments, was 33.33% and was 5 times higher than treatment ② and treatment ④, treatment ② had the lowest callus browning rate, but induced callus redifferentiation treatment ⑤, and similar results were also found on crown eriobotrya japonica, indicating that the plants IAA regenerated by inducing differentiation of the loquat callus were better than NAA, and the present experiment resulted in the final production of 28 white eriobotrya japonica tissue culture seedlings, more than 60% of those obtained by induction with ⑤, treatment ⑥ with 6-BA (2mg/L) added alone, resulted in moderate statistical performance of anti-callus browning and callus greening, and only a few tissue culture seedlings were obtained, unlike 2mg/LZT +0.2mg/⑤.
2.2 multiplication and rooting of shoots
When two small leaves grow from the buds of the callus of the white jade and the Guanyu loquat, the buds can be cut and transplanted, as shown in figure 2A, the buds are inoculated into a culture medium of MS + ZT (2mg/L), the growth state is good, and the tillering of the stem bottom is obvious about two months (as shown in figure 2B).
After subculture, the newly cut tissue culture seedlings are inoculated into a rooting culture medium for culture, the rooting rate reaches 50% after one month, and the rooting rate reaches 83.33% after two months. As shown in FIG. 3, the seedlings inoculated into the rooting medium were observed to grow well, indicating that the 1/2MS + IBA (0.4mg/L) medium was suitable for rooting of loquat seedlings.
In conclusion, the research provides a rapid breeding system for inducing callus and differentiating and regenerating loquat by using young loquat embryos. The specific operation is that in the middle and last ten days of 3 months in Jiangsu area, the transverse diameter of young loquat fruits is 1.0-1.5 cm, the low-temperature pretreatment is carried out for 15 days at 4 ℃, sepals at the bottoms of the fruits and fuzz on the surfaces of the fruits are removed, the fruits are washed for four hours by running water, the surfaces of the fruits are disinfected by 70% alcohol for 10min, then disinfected by 30% sodium hypochlorite for 15min, and washed by sterile water for 3-4 times. The young embryo of the loquat young fruit is stripped on an ultraclean workbench and inoculated on a callus induction culture medium which is added with 2mg/L6-BA and 0.5 mg/L2, 4-D in MS basic culture medium, and the average induction rate of the callus is 51.11% (white jade) and 66.67% (crown jade). The callus is transferred to a bud regeneration culture medium added with 2mg/L ZT +0.2mg/L IAA in an MS culture medium, and the regeneration rate of the callus can be more than 80 percent. Culturing the regenerated bud on MS culture medium containing 2mg/L ZT to promote rapid growth and tillering, and subculturing. The regenerated seedlings were rooted on a rooting medium supplemented with 0.4mg/L IBA in 1/2MS basal medium. The invention overcomes the problems of easy browning and difficult explant sterilization in the past by utilizing stem tip culture, and greatly improves the propagation efficiency.
Claims (5)
1. A tissue culture and rapid propagation method of white sand loquat seedlings is characterized by comprising the following steps: picking young loquat fruits with the transverse diameter of 10-12 mm and green peel but yellow villus in the middle and last ten days of 4 months in Jiangsu area, pretreating at the low temperature of 4 ℃ for 12-15 days, removing sepals at the bottoms of the fruits, removing the hairy hairs on the surfaces of the fruits, disinfecting and cleaning, stripping young embryos of the young loquat fruits on a clean bench, inoculating the young loquat fruits on an MS basic culture medium with 2mg/L6-BA and 0.5 mg/L2, 4-D callus induction culture medium, and carrying out dark culture at the temperature of 23-25 ℃ for 45-50 days to induce the formation of callus; subculturing the callus for 1 time until the callus grows to 1cm2Inoculating the culture medium to a bud regeneration culture medium at 23-25 ℃ for culturing for 55-60 days to induce the formation of buds, wherein the bud regeneration culture medium is an MS culture medium added with 2mg/L ZT +0.2mg/L IAA; when two leaflets grow out from the buds of the callus, cutting the buds and transplanting the buds to an MS culture medium containing 2mg/L ZT for subculture for 55-65 days at 23-25 ℃, inoculating the newly cut tissue culture seedlings after subculture to 1/2MS basal medium, adding 0.4mg/L IBA rooting culture medium, and culturing for 30-35 days at 23-25 ℃ to obtain the white sand loquat seedlings.
2. The method as claimed in claim 1, wherein young loquat fruits with cross diameter of 10 mm-12 mm and green pericarp but still yellow villus are picked up at 3 months 20-25 days or 4 months 8-10 days in Jiangsu area, pretreated at 4 ℃ for 15 days, sepals at the bottom of the fruits, hairy hairs on the surface of the fruits are removed, the fruits are washed with running water for four hours, surface disinfection is carried out with 70% alcohol for 10min, then disinfection is carried out with 30% sodium hypochlorite for 15min, and the fruits are washed with sterile water for 3-4 times. And stripping the young embryo of the loquat young fruit on a clean bench.
3. The method as claimed in claim 1, wherein the loquat young fruit is pretreated at 4 ℃ for 15 days.
4. The method according to claim 1, wherein the young embryo is inoculated on the callus induction medium and cultured in dark at 23 ℃ for 50 days to induce callus formation.
5. The method of claim 1, wherein the culture light intensity is 3000-5000 Lx except for callus induced in dark.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN102994443A (en) * | 2012-11-14 | 2013-03-27 | 福建省亚热带植物研究所 | Suspension culture solution and suspension culture method for loquat cells |
| CN103404442A (en) * | 2013-08-15 | 2013-11-27 | 湖南农业大学 | Induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther |
| CN104404156A (en) * | 2014-12-08 | 2015-03-11 | 南京农业大学 | Rapid identification molecular marker of self-compatible variety of loquat, marker primer and identification method |
| CN106508671A (en) * | 2016-11-16 | 2017-03-22 | 四川农业大学 | Method for breeding earlier germinated eriobotrya japonica seeds |
| WO2018220181A1 (en) * | 2017-06-02 | 2018-12-06 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Method for automated transformation of a plant cell pack |
-
2019
- 2019-11-05 CN CN201911071911.2A patent/CN110710452A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| WO2005012507A1 (en) * | 2003-07-25 | 2005-02-10 | The University Of Melbourne | Production of plant secondary metabolites using adsorption and elicitation in cell suspension culture |
| CN102994443A (en) * | 2012-11-14 | 2013-03-27 | 福建省亚热带植物研究所 | Suspension culture solution and suspension culture method for loquat cells |
| CN103404442A (en) * | 2013-08-15 | 2013-11-27 | 湖南农业大学 | Induction method for embryogenesis and plant regeneration of in vitro haploid of macleaya cordata anther |
| CN104404156A (en) * | 2014-12-08 | 2015-03-11 | 南京农业大学 | Rapid identification molecular marker of self-compatible variety of loquat, marker primer and identification method |
| CN106508671A (en) * | 2016-11-16 | 2017-03-22 | 四川农业大学 | Method for breeding earlier germinated eriobotrya japonica seeds |
| WO2018220181A1 (en) * | 2017-06-02 | 2018-12-06 | Fraunhofer-Gesellschaft zur Förderung der angewandten Forschung e.V. | Method for automated transformation of a plant cell pack |
Non-Patent Citations (4)
| Title |
|---|
| MANUEL BLASCO等: ""Embryogenic response from anther culture of cultivars"", 《EUPHYTICA》 * |
| 于丽杰等: "《植物组织培养教程》", 31 August 2015, 华中科技大学出版社 * |
| 徐海红: ""枇杷组织培养及SS基因生物信息学分析"", 《中国优秀硕士学位论文全文数据库（电子期刊） 农业科技辑》 * |
| 范晨昕: ""白沙枇杷离体再生及ISSR分子标记研究"", 《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》 * |
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