CN105123514A - Induction method for initial culture bud of Kiwi berry - Google Patents
Induction method for initial culture bud of Kiwi berry Download PDFInfo
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- CN105123514A CN105123514A CN201510447705.2A CN201510447705A CN105123514A CN 105123514 A CN105123514 A CN 105123514A CN 201510447705 A CN201510447705 A CN 201510447705A CN 105123514 A CN105123514 A CN 105123514A
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Abstract
The invention provides an induction method for an initial culture bud of Kiwi berry. The method comprises the following steps: (1) selection of explant: a step of selecting an annual branch of Kiwi berry as explant, scissoring the branch with a pair of scissors and inserting the branch in water to prevent evaporation of moisture; (2) disinfection of the explant: a step of scissoring leaves on the branch of Kiwi berry obtained in the step (1), carrying out primary trimming and cleaning, transferring the branch to an aseptic environment, subjecting the branch to disinfection and flushing and then carrying out trimming again so as to obtain stem segments; (3) inoculation: a step of inoculating the stem segments on bud induction mediums, wherein each glass bottle is inoculated with one stem segment; and (4) bud induction culture: a step of putting the inoculated explant in a culture room, carrying out culture under the condition of dim light at first and then carrying out culture under the condition of high light. Compared with the prior art, the invention has the following advantages: with the induction method and through special treatment, the survival rate and the germination rate of the explant are substantially increased, and the phenomenon of browning is prevented.
Description
Technical field
The present invention relates to a kind of kiwi fruit Initial culture bud inducement method, belong to kiwi fruit propagation technique field.
Background technology
Kiwi fruit is the wild fruit tree of artificial domesticating cultivation, and claim monkey Peach fruits to contain abundant nutrient component, abundant vitamin, mineral element and 14 seed amino acids, in every 1009 fresh fruit meat, Vc reaches 90-420mg, enjoys the laudatory title of " king of vitamin C ".Kiwi fruit can be eaten raw, also can be processed into jam fruit juice etc., and also having medical care effect, is the fruit be loved by the people.Red kiwi fruit belongs to Chinese gooseberry series, and be the natural variation kind found in Cangxi, Cheng Houtao original producton location, Sichuan, early fruiting character, yielding ability and strong stress resistance, have high promotion prospect, and the proterties of its red meat has value of exploiting and utilizing.
Kiwi fruit is dioecism, and adopt seminal propagation, the cycle is long, and not easily obtains large quantities of female plant colony, and utilizes cuttage, grafting, and survival rate is low, and efficiency is low.Adopt conventional method can not meet require that of large-scale planting.Thus tissue culture technique breeding kiwi fruit is adopted to become important stock breeding approach.In Kiwifruit Tissue Culture, there is genotypic difference in different kinds, and different cultivars needs different medium.In the past in the research of red kiwi fruit, be that minimal medium is studied with MS, and showed that inducing clumping bud rate is the highest and only have 27.27%, greatly have impact on the foundation of kiwi fruit rapid propagation system, constrain the large-scale production of red kiwi fruit.
Therefore, current kiwi fruit inducing clumping bud technology, survival rate and germination rate too low, and melting brown rate is too high, has had a strong impact on kiwi fruit young shoot quality, cannot meet the needs of shoot proliferation.
Summary of the invention
Goal of the invention: in order to overcome the deficiencies in the prior art, the invention provides a kind of kiwi fruit Initial culture bud inducement method.
Technical scheme: for achieving the above object, the invention provides a kind of kiwi fruit Initial culture bud inducement method, comprises the following steps:
(1) explant is selected: select kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: cut by step (1) gained kiwi fruit branch blade, then just prunes, cleans, then forward in gnotobasis, sterilization, rinse, then prune, obtain stem section;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, first cultivates under low light condition, then cultivates under intense light conditions.
As preferably, just to prune in described step (2) and the method for cleaning is: the rectangular of described branch is first cut into 5-8cm long shoot bar, through washing powder cleaning, last running water 2-4h.
Preferred as another kind, sterilization in described step (2), the method for rinsing and pruning are: first used for 75% alcohol disinfecting 20-30 second, then with 0.1% mercuric chloride sterilization 6-8 minute, then after using aseptic water washing 8-10 time, are cut into the stem section of 1-2cm length.
Preferred as another kind, in described step (3), medium is WPM minimal medium, and it also comprises following composition: 6-benzylaminopurine 0.5-2mg/L, methyl α-naphthyl acetate 0.1-1mg/L, active carbon 2-5mg/L, vitamin C 5-10mg/L, agar 7g/L, sucrose 30g/L.
Preferred as another kind, in described step (3), medium regulates pH to be 5.8.
Preferred as another kind, in described step (4), low light condition is: light intensity is 800-1000lux, and incubation time is 7 days.
Preferred as another kind, in described step (4), intense light conditions is: light intensity is 2500-3000lux, and incubation time is 15-25 days.
Preferred as another kind, in described step (4) culturing room, cultivation temperature is 25 scholar 2 DEG C.
Preferred as another kind, described kiwi fruit is red kiwi fruit.
The minimal medium of WPM described in the present invention is that medium is commonly used in this area, on the books in prior art.
Beneficial effect: relative to prior art, kiwi fruit Initial culture bud inducement method of the present invention, not only there is the advantage that stabilization characteristics of genetics and incubation time in prior art are short, the most important thing is by whole method, and special process, substantially increase explant survival rate and germination rate, it also avoid simultaneously and occur browning.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
Embodiment 1
(1) explant is selected: select red kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: step (1) gained kiwi fruit branch blade is cut, then the rectangular of branch is first cut into 5-8cm long shoot bar, clean through washing powder, last running water 2h, forward to again on aseptic operating platform, first use 75% alcohol disinfecting 20 seconds, then sterilize 6 minutes with 0.1% mercuric chloride, after using aseptic water washing 8 times again, be cut into the stem section that 1-2cm is long;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section, described medium is WPM minimal medium, it also comprises following composition: 6-benzylaminopurine 0.5mg/L, methyl α-naphthyl acetate 0.1mg/L, active carbon 2mg/L, vitamin C 5mg/L, agar 7g/L, sucrose 30g/L, regulates medium pH to be 5.8;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, cultivation temperature is 25 scholar 2 DEG C, first cultivates under light intensity is the low light condition of 800lux, then in light intensity be 2500lux intense light conditions under cultivate 15 days.
Embodiment 2
(1) explant is selected: select red kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: step (1) gained kiwi fruit branch blade is cut, then the rectangular of branch is first cut into 5-8cm long shoot bar, clean through washing powder, last running water 4h, forward to again on aseptic operating platform, first use 75% alcohol disinfecting 30 seconds, then sterilize 8 minutes with 0.1% mercuric chloride, after using aseptic water washing 10 times again, be cut into the stem section that 1-2cm is long;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section, described medium is WPM minimal medium, it also comprises following composition: 6-benzylaminopurine 2mg/L, methyl α-naphthyl acetate 1mg/L, active carbon 5mg/L, vitamin C 10mg/L, agar 7g/L, sucrose 30g/L, regulates medium pH to be 5.8;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, cultivation temperature is 25 scholar 2 DEG C, first cultivates under light intensity is the low light condition of 1000lux, then in light intensity be 3000lux intense light conditions under cultivate 25 days.
Embodiment 3
(1) explant is selected: select red kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: step (1) gained kiwi fruit branch blade is cut, then the rectangular of branch is first cut into 5-8cm long shoot bar, clean through washing powder, last running water 3h, forward to again on aseptic operating platform, first use 75% alcohol disinfecting 25 seconds, then sterilize 7 minutes with 0.1% mercuric chloride, after using aseptic water washing 9 times again, be cut into the stem section that 1-2cm is long;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section, described medium is WPM minimal medium, it also comprises following composition: 6-benzylaminopurine 1.2mg/L, methyl α-naphthyl acetate 0.5mg/L, active carbon 3mg/L, vitamin C 8mg/L, agar 7g/L, sucrose 30g/L, regulates medium pH to be 5.8;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, cultivation temperature is 25 scholar 2 DEG C, first cultivates under light intensity is the low light condition of 900lux, then in light intensity be 2700lux intense light conditions under cultivate 20 days.
Embodiment 4
(1) explant is selected: select red kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: step (1) gained kiwi fruit branch blade is cut, then the rectangular of branch is first cut into 5-8cm long shoot bar, clean through washing powder, last running water 3h, forward to again on aseptic operating platform, first use 75% alcohol disinfecting 22 seconds, then sterilize 7 minutes with 0.1% mercuric chloride, after using aseptic water washing 8 times again, be cut into the stem section that 1-2cm is long;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section, described medium is WPM minimal medium, it also comprises following composition: 6-benzylaminopurine 0.8mg/L, methyl α-naphthyl acetate 0.3mg/L, active carbon 3mg/L, vitamin C 7mg/L, agar 7g/L, sucrose 30g/L, regulates medium pH to be 5.8;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, cultivation temperature is 25 scholar 2 DEG C, first cultivates under light intensity is the low light condition of 850lux, then in light intensity be 2600lux intense light conditions under cultivate 17 days.
Embodiment 5
(1) explant is selected: select red kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: step (1) gained kiwi fruit branch blade is cut, then the rectangular of branch is first cut into 5-8cm long shoot bar, clean through washing powder, last running water 4h, forward to again on aseptic operating platform, first use 75% alcohol disinfecting 28 seconds, then sterilize 6 minutes with 0.1% mercuric chloride, after using aseptic water washing 9 times again, be cut into the stem section that 1-2cm is long;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section, described medium is WPM minimal medium, it also comprises following composition: 6-benzylaminopurine 1.8mg/L, methyl α-naphthyl acetate 0.8mg/L, active carbon 4mg/L, vitamin C 8mg/L, agar 7g/L, sucrose 30g/L, regulates medium pH to be 5.8;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, cultivation temperature is 25 scholar 2 DEG C, first cultivates under light intensity is the low light condition of 950lux, then in light intensity be 2900lux intense light conditions under cultivate 22 days.
Experimental example each method gained explant survival rate, germination rate and melting brown rate result
The red kiwi fruit choosing same batch carries out Initial culture, be respectively contrast 1 group, contrast 2 groups, embodiment 3 groups, embodiment 4 groups and embodiment 5 groups.
Contrast 1 group, according to the method for the embodiment of the present invention 3, medium is replaced with conventional MS medium, other are constant cultivates;
Contrast 2 groups, according to the method for the embodiment of the present invention 3, be low light condition 900lux unlike illumination cultivation condition in step (4);
Contrast 3 groups, according to the method for the embodiment of the present invention 3, be intense light conditions 2700lux unlike illumination cultivation condition in step (4);
Calculate the survival rate of each group, germination rate and melting brown rate, the results are shown in Table 1.
Table 1 each method gained explant survival rate, germination rate and melting brown rate result
Can be obtained by upper table 1 result, contrast 1 group of comparing, namely use conventional MS medium, survival rate and the germination rate of the inventive method gained explant are significantly increased, and melting brown rate is 0, browning does not occur;
In addition, compare contrast 2 groups and contrast 3 groups, survival rate and the germination rate of the inventive method gained explant are also significantly increased, and melting brown rate is 0, browning does not occur.
Claims (9)
1. a kiwi fruit Initial culture bud inducement method, is characterized in that, comprise the following steps:
(1) explant is selected: select kiwi fruit current-year branch to be explant, is inserted in water in case moisture evaporation with scissors after cutting;
(2) explant sterilization: cut by step (1) gained kiwi fruit branch blade, then just prunes, cleans, then forward in gnotobasis, sterilization, rinse, then prune, obtain stem section;
(3) inoculate: above-mentioned stem section is inoculated on bud inducement medium, every glass bottle graft stem section;
(4) bud inducement is cultivated: be placed in culturing room by postvaccinal explant, first cultivates under low light condition, then cultivates under intense light conditions.
2. kiwi fruit Initial culture bud inducement method according to claim 1, it is characterized in that, the method of just pruning in described step (2) and clean is: the rectangular of described branch is first cut into 5-8cm long shoot bar, through washing powder cleaning, and last running water 2-4h.
3. kiwi fruit Initial culture bud inducement method according to claim 1, it is characterized in that, sterilization in described step (2), the method for rinsing and pruning are: first used for 75% alcohol disinfecting 20-30 second, again with 0.1% mercuric chloride sterilization 6-8 minute, after using aseptic water washing 8-10 time again, be cut into the stem section that 1-2cm is long.
4. kiwi fruit Initial culture bud inducement method according to claim 1, it is characterized in that, in described step (3), medium is WPM minimal medium, and it also comprises following composition: 6-benzylaminopurine 0.5-2mg/L, methyl α-naphthyl acetate 0.1-1mg/L, active carbon 2-5mg/L, vitamin C 5-10mg/L, agar 7g/L, sucrose 30g/L.
5. kiwi fruit Initial culture bud inducement method according to claim 1, is characterized in that, in described step (3), medium regulates pH to be 5.8.
6. kiwi fruit Initial culture bud inducement method according to claim 1, is characterized in that, in described step (4), low light condition is: light intensity is 800-1000lux, and incubation time is 7 days.
7. kiwi fruit Initial culture bud inducement method according to claim 1, is characterized in that, in described step (4), intense light conditions is: light intensity is 2500-3000lux, and incubation time is 15-25 days.
8. kiwi fruit Initial culture bud inducement method according to claim 1, is characterized in that, in described step (4) culturing room, cultivation temperature is 25 scholar 2 DEG C.
9. kiwi fruit Initial culture bud inducement method according to claim 1, it is characterized in that, described kiwi fruit is red kiwi fruit.
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| CN201510447705.2A CN105123514A (en) | 2015-07-27 | 2015-07-27 | Induction method for initial culture bud of Kiwi berry |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105454049A (en) * | 2016-01-04 | 2016-04-06 | 江苏农林职业技术学院 | Kiwi fruit multiplication medium and preparing method and application thereof |
| CN108522277A (en) * | 2018-03-13 | 2018-09-14 | 贵州省山地资源研究所 | A kind of tissue culture method of " Red Male " Kiwi berry semi-lignified stem with bud |
-
2015
- 2015-07-27 CN CN201510447705.2A patent/CN105123514A/en active Pending
Non-Patent Citations (3)
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| 张弛: "红阳猕猴桃四倍体诱导及其抗溃疡病特性初探", 《中国优秀硕士学位论文全文数据库 农业科技辑》 * |
| 王俊丽: "《细胞工程原理与技术》", 31 December 2006, 中央民族大学出版社 * |
| 王洪习等: "《植物组织培养技术》", 31 January 2014, 机械工业出版社 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105454049A (en) * | 2016-01-04 | 2016-04-06 | 江苏农林职业技术学院 | Kiwi fruit multiplication medium and preparing method and application thereof |
| CN108522277A (en) * | 2018-03-13 | 2018-09-14 | 贵州省山地资源研究所 | A kind of tissue culture method of " Red Male " Kiwi berry semi-lignified stem with bud |
| CN108522277B (en) * | 2018-03-13 | 2021-08-27 | 贵州省山地资源研究所 | Tissue culture seedling raising method for semi-lignified stem segments with buds of 'Hongyang' kiwi fruits |
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Application publication date: 20151209 |