CN108575756A - A kind of method of morning pears shoot tip in vitro culture - Google Patents
A kind of method of morning pears shoot tip in vitro culture Download PDFInfo
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- CN108575756A CN108575756A CN201810445954.1A CN201810445954A CN108575756A CN 108575756 A CN108575756 A CN 108575756A CN 201810445954 A CN201810445954 A CN 201810445954A CN 108575756 A CN108575756 A CN 108575756A
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- pears
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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 method of early pears shoot tip in vitro culture：Early pears stem apex is cultivated successively with four kinds of culture mediums containing graphene quantum dot, after 180 270d, complete healthy and strong test tube seedling can be formed.Compared with the prior art, early pears stem apex browning substantially reduces the present invention, survival rate is high, and the test tube seedling detoxification efficiency formed is good, and robust growth is easy to transplant.
Description
Technical field：
The present invention relates to a kind of methods of plant shoot tip in vitro culture, are exactly a kind of early pears shoot tip in vitro culture
Method.
Background technology：
Shangrao morning pears originate from Woody plants.Shangrao County is a Mountain Area County based on hills, and no substantial industrial is dirty
Dye source, ecological environment is excellent, is very suitable for plantation Shangrao morning pears.The big thin skin of Shangrao morning the operatic circle, color white light are slided, fine and tender taste, juice
More slags are few, sweet slightly sour, and palatable crisp is not overnight yellow, and containing a variety of battalion such as minerals, carrotene and vitamin B, vitamin C
Element is supported, there is promoting production of body fluid and nourishing the lung, clearing heat and eliminating phlegm, antipyretic relieve summer heat and other effects.According to《Xinzhou mansion will》It records, Shangrao morning pears of reception room or parlour production
Early in the Qing Dynasty, just the upper tribute imperial palace with local speciality, cultivation history had for more than 400 years.Serissa serissoide, Calusena lansium such as disappear at the main local product
Kind be referred to as Shangrao morning pears, especially using serissa serissoide as Shangrao the operatic circle most, 2005, in Jiangxi, first Gan Bei morning pears sections obtained gold medal,
2009, " ten big fine fruit of East China " title is obtained in Chinese (Quzhou) agriculture exposition.2012, Shangrao morning pears were granted for country
Agricultural product geography symbol product.Currently, really Shangrao morning pears (such as serissa serissoide and Calusena lansium disappear) rare cultivation, mostly tens
Year so century-old veteran, only the Shangrao County reception room or parlour town towns He Tiandun some areas also have a small amount of cultivation, but that there are virus diseases is serious,
The problems such as fruit yield declines, storage tolerance, kind sexual involution, quality do not deteriorate, the even whole strain of some is dead and has no harvest, and seriously affects
Shangrao morning pears production.In addition, virosis has an effect on Shangrao morning pears seedling quality, cause anvil fringe not affine and seedling root development
It is bad, and band poison throughout one's life, persistently cause harm.Virus type disease has become the serious hindrance of the excellent production of Shangrao morning pears high yield, should try
It is inherently eliminated causing harm for virosis.Shoot-tip Grafting In Vitro is one of most important, most active field in agricultural high-tech,
It is not only the basis of agricultural sustainable development, and is most widely used, most realistic meaning field in biotechnology, is praised
For the 4th green revolution in agriculture development history.Domestic and international many results of study, which are proved stem tip tissue culture, certain take off
Toxic effect fruit, Shoot Tip Culture detoxification principle be using virus in plant nonunf ormity, closer to stem top meristematic region,
Virus concentration is also lower, i.e., the separate living tissue of the tip of a root and bud point containing virus quantity it is few or without virus.Therefore, according to stem apex, there are nothings
Viral area aseptically cuts plant stem apex and carries out cultured in vitro, can obtain not viruliferous plant.Stem apex detoxification is trained
Two foster key techniques are the survival rate and virus elimination rate of stem apex, browning be reduce Shoot Tip Culture survival rate it is primary because
Element.The generation of browning is related with many factors, stem apex size, materials season, training method, hormone concentration, minimal medium, training
It supports base additive etc. all to have an impact stem apex browning, such as stem apex is smaller, browning is more serious, and survival rate is lower.It is asked for these
Topic, the present invention develop a kind of method of early pears shoot tip in vitro culture, can be that the large-scale production of Shangrao morning pears detoxic seedling is long-term
Technical foundation is provided.
Invention content：
The object of the present invention is to provide a kind of browning reduction, the early pears shoot tip in vitro culture that survival rate is high, detoxification efficiency is good
Method.Realizing the technical solution of the object of the invention is, a kind of method of morning pears shoot tip in vitro culture, it is characterised in that have following
Step：(1) in superclean bench, 0.2-0.3mm morning pears stem apexs is cut and are inoculated in one (MS+0.3-0.5g/ of culture medium
L graphene quantum dot+30g/L sucrose+0g/L agar), material is put into culturing room after inoculation and carries out normal condition dark culturing,
The temperature of culturing room is 25 ± 2 DEG C；(2) after cultivating 7-10d on culture medium one, early pears stem apex is transferred to two (MS+1- of culture medium
2mg/L Thidiazuron+0.1-0.3mg/LNAA+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar), inoculation
Material is put into culturing room afterwards and carries out normal condition culture, culturing room's condition of culture is：25 ± 2 DEG C of temperature, light application time 16h/
D, light intensity 1500-2500lx；(3) after cultivating 60-90d on culture medium two, early pears simple bud is transferred to culture medium three：MS+1-
2mg/L Thidiazuron+0.1-0.3mg/LNAA+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar, after inoculation
Material is put into culturing room and carries out normal condition culture；(4) after cultivating 60-90d on culture medium three, early pears adventitious bud is transferred to
Culture medium four (1/2MS+0.5-1mg/L NAA+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar), connects
Material is put into culturing room after kind and carries out normal condition culture, culturing room's condition of culture is：25 ± 2 DEG C of temperature, light application time
16h/d, light intensity 1500-2500lx；(5) after cultivating 60-90d on culture medium four, early pears adventitious bud rooting is formed complete strong
Strong test tube seedling.
Specific implementation mode：
In conjunction with following embodiments, the invention will be further described：
(1) it cultivates for the first time
In superclean bench, cuts 0.2-0.3mm morning pears stem apexs and be inoculated in one (MS+0.3-0.5g/ of culture medium
L graphene quantum dot+30g/L sucrose+0g/L agar), material is put into culturing room after inoculation and carries out normal condition dark culturing,
The temperature of culturing room is 25 ± 2 DEG C；
(2) it cultivates for second
After cultivating 7-10d on culture medium one, early pears stem apex is transferred to the (MS+1-2mg/L Thidiazurons+0.1- of culture medium two
0.3mg/LNAA+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar), material is put into culture after inoculation
Room carries out normal condition culture, and culturing room's condition of culture is：25 ± 2 DEG C, light application time 16h/d, light intensity 1500- of temperature
2500lx；
(3) third time is cultivated
After cultivating 60-90d on culture medium two, early pears simple bud is transferred to culture medium three：MS+1-2mg/L Thidiazurons+0.1-
0.3mg/LNAA+0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar, culturing room is put into after inoculation by material
Carry out normal condition culture；
(4) the 4th cultures
After cultivating 60-90d on culture medium three, early pears adventitious bud is transferred to (the 1/2MS+0.5-1mg/L NAA of culture medium four
+ 0.3-0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar), material is put into culturing room after inoculation and carries out routine
CMC model, culturing room's condition of culture are：25+2 DEG C of temperature, light application time 16h/d, light intensity 1500-2500lx；
After cultivating 60-90d on culture medium four, early pears adventitious bud rooting forms complete healthy and strong test tube seedling.
Claims (1)
1. the method for early pears shoot tip in vitro culture, it is characterised in that there is following steps：(1) in superclean bench, 0.2- is cut
0.3mm morning pears stem apexs are simultaneously inoculated in culture medium one：MS+0.3-0.5g/L graphene quantum dot+30g/L sucrose+0g/L fine jades
Material is put into culturing room after inoculation and carries out normal condition dark culturing by fat；It (2), will be early after cultivating 7-10d on culture medium one
Pears stem apex is transferred to culture medium two：MS+1-2mg/L Thidiazuron+0.1-0.3mg/LNAA+0.3-0.5g/L graphene quantum dots+
Material is put into culturing room after inoculation and carries out normal condition culture by 30g/L sucrose+7.5g/L agar；(3) it is trained on culture medium two
After supporting 60-90d, early pears simple bud is transferred to culture medium three：MS+1-2mg/L Thidiazurons+0.1-0.3mg/LNAA+0.3-0.5g/L
Material is put into culturing room after inoculation and carries out normal condition culture by graphene quantum dot+30g/L sucrose+7.5g/L agar；(4)
After cultivating 60-90d on culture medium three, early pears adventitious bud is transferred to culture medium four：1/2MS+0.5-1mg/L NAA+0.3-
Material is put into culturing room after inoculation and carries out normal condition training by 0.5g/L graphene quantum dot+30g/L sucrose+7.5g/L agar
It supports；(5) after cultivating 60-90d on culture medium four, early pears adventitious bud rooting forms complete healthy and strong test tube seedling.
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| CN201810445954.1A CN108575756A (en) | 2018-05-03 | 2018-05-03 | A kind of method of morning pears shoot tip in vitro culture |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109452156A (en) * | 2018-11-28 | 2019-03-12 | 上饶师范学院 | A method of improving early pears stem apex detoxification efficiency |
Citations (3)
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|---|---|---|---|---|
| CN102648698A (en) * | 2012-05-23 | 2012-08-29 | 南京农业大学 | Pyrus stem tip tissue culture rapid propagation method |
| CN105794642A (en) * | 2016-04-01 | 2016-07-27 | 南京农业大学 | Method for efficiently and rapidly regenerating adventitious buds from pear leaves |
| CN107853176A (en) * | 2017-11-14 | 2018-03-30 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method of the aseptic culture medium containing carbon nanomaterial |
-
2018
- 2018-05-03 CN CN201810445954.1A patent/CN108575756A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102648698A (en) * | 2012-05-23 | 2012-08-29 | 南京农业大学 | Pyrus stem tip tissue culture rapid propagation method |
| CN105794642A (en) * | 2016-04-01 | 2016-07-27 | 南京农业大学 | Method for efficiently and rapidly regenerating adventitious buds from pear leaves |
| CN107853176A (en) * | 2017-11-14 | 2018-03-30 | 中国烟草总公司郑州烟草研究院 | A kind of preparation method of the aseptic culture medium containing carbon nanomaterial |
Non-Patent Citations (5)
| Title |
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| W. DAVID LANE: "Shoot regeneration from cultured leaves of Japanese pear (Pyrus pyrifolia)", 《PLANT CELL,TISSUE AND ORGAN CULTURE》 * |
| 宗娟等: "黄冠梨茎尖离体培养再生体系建立初探", 《广东农业科学》 * |
| 崔瑾等: "苯基噻二唑基脲（TDZ）和琼脂对芋茎尖生长的影响", 《江苏农业科学》 * |
| 赵惠祥: "梨茎尖离体培养", 《植物学报》 * |
| 赵振杰等: "碳纳米材料对植物生长发育的调节作用", 《作物杂志》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109452156A (en) * | 2018-11-28 | 2019-03-12 | 上饶师范学院 | A method of improving early pears stem apex detoxification efficiency |
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