CN104381132A - Rooting method of Juglans hindsii*J.regia vlach tissue culture seedling in bottle - Google Patents
Rooting method of Juglans hindsii*J.regia vlach tissue culture seedling in bottle Download PDFInfo
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Abstract
The invention discloses a rooting method of a Juglans hindsii*J.regia vlach tissue culture seedling in a bottle. A healthy Juglans hindsii*J.regia vlach tissue culture seedling in a subculture period is used as a rooting material and is treated by dark culture; then the Juglans hindsii*J.regia vlach tissue culture seedling is conveyed into a rooting culture medium to be subjected to rooting culture with alternative light culture and dark culture; a tissue culture rooting seedling which is planted in the bottle can be obtained after 20-24 days; and the rooting rate is up to 65%-75% and the greenhouse survival rate is up to 65%-85%. The rooting method of the Juglans hindsii*J.regia vlach tissue culture seedling in the bottle disclosed by the invention is not influenced by factors including seasons and the like, and theoretical basis and technical supports are provided for industrialized production, and uniform and stable germchits can be efficiently and rapidly provided; and the popularization speed of improved variety is accelerated. The rooting method is applicable to rooting culture of the Juglans hindsii*J.regia vlach tissue culture seedling in the bottle.
Description
Technical field
The invention belongs to domestication of plants field, relate to a kind of plant establishment method, be specifically related to the rooting method in a kind of Paradox walnut crow clarke plantlet in vitro bottle.
Background technology
Walnut is a kind of nonwood forest trees being difficult to take root, and there were significant differences for different IPs Peach cultivars rooting characteristics.According to California, USA university Gale McGranahan 2004, in the holy cold spy (Sunlan) of 3 walnut main breeds, by force Teller (Chandler), Wei Na (Vina) that laboratory adopts, situation of taking root is widely different, growth potential is strong, the upright kind of tree performance holy cold spy the most easily take root, and rooting rate reaches 94%; Strong Teller rootability is in middle level, reaches 55%; Wei Na is then the most difficult kind of taking root, and rooting rate only has 27%.This research also confirms that Walnut Cultivars or genotype are the key factors determining the raw rooting of vitro seedling of walnut.
" progress of walnut rooting of vitro seedling " Li Chunyan etc., " northern gardening ", Walnut Cultivars difference is disclosed in 2013 (01): 190-193 pages, genotype is different, also there is larger difference in the expression of its rootability or potential of taking root, shows the capacity variance great disparity that adventive root occurs between different tree species and kind individuality.
" impact that IBA and dark treatment are taken root on walnut plantlet in vitro " Li Chunyan etc., " Shanxi Forestry science and technology ", in December, 2011,40th volume the 4th phase, 30-31 page, with " gold rich fragrant No. 6 " walnut plantlet in vitro for examination material, has inquired into the impact that IBA concentration and dark treatment number of days are taken root on walnut plantlet in vitro, under its optimum condition, rooting rate can reach 65.56%.And point out: different growth hormone is different to walnut root induction effect.Adventive root production process not only depends on the content of Endogenous auxin, also relates to the balance between hormone.
" J. sigillata cultured in vitro and Fast-propagation " Wang Shasha etc., " plant physiology journal ", 2014,50 (4), 527-537 page, to disclose with the stem-segment with single bud of fine individual plant " Nayong-1 " as explant, establishes the system of J. sigillata (Juglanssigillata) cultured in vitro and Fast-propagation, its the highest rooting rate is 71.73%
And Paradox walnut (Juglans hindsii × J.regia) belongs to Juglandaceae (Juglandales) juglans (Juglans), it is the crossbreed of U.S.'s north California black walnut and walnut, wherein male parent is walnut, and female parent is northern California black walnut.This seeds crow clarke Vlach is the elite plant strain selected from Paradox walnut, it is a walnut stock variety, introduced from California, USA in 2010 through prominent domestic walnut expert Xi Shengke professor, it has obvious hybrid vigour, obviously be better than existing walnut stock, growth potential is good, has anti-nematode, Phytophthora germ, rhizobium, the disease that halimasch etc. cause.Bioclimatic analysis is wide, can on the soil of pH > 5 normal growth, normal growth on the calcium soil of pH=9.This kind has certain cold resistance, and suitable growth is in the abundant area of rain profit, and the region of rainfall > 500mm all can normal growth.Because its growing way is prosperous, be particularly suitable as the stock of early-fruiting walnut, strengthen tree vigo(u)r.Do stock with it, the just full bearing period within general 6 years, can be reached.Meanwhile, these seeds are the hard wealthy arbor of fallen leaves, and growth potential is strong, and trunk is logical straight, be the excellent material of urban greening and the greening of farmland woods, compared with black walnut, its material is slightly shallow, between black walnut and walnut, can be used as hard wealthy material and use, this type of timber many genus import domestic at present.
" research of Paradox walnut fast breeding technique " Qiu Xiaomei etc., " Inner Mongol forestry science and technology ", the 39th volume, 2nd phase, 52-54 page, discloses with Paradox walnut stem segment with axillary bud for explant carries out Propagations of Teat Tube Seedlings, and axillalry bud germinating medium and differentiation, squamous subculture are DKW+IBA 1.0mg/L.Use " two step method " root induction effect better, the suitableeest root induction medium is DKW+IBA 2.5mg/L, and root media is the DKW without plant hormone, and rooting rate can reach 65%.Because Paradox walnut belongs to crossbreed, the Paradox walnut seedling gone out of each seed culture all belongs to different genotype.
And Paradox walnut is scion on domestic market, strain source is chaotic, there is no clone and to take root seedling.Paradox walnut (vlach) branch cutting rooting rate is low, and Plant Tissue Breeding, as vegetative manner, becomes the inevitable mode of being carried out by this choiceness breeding, but due to its difficult problem of taking root, not yet forms industrialization at present.Current nursery stock markets coker Peach rootstock is various in style, very different, and mostly is planting seed and obtains, there is the problem of trait segregation in offspring, merit is difficult to genetic stability, though have the report of walnut tissue-culturing rapid propagation in existing research, but because its difficulty is taken root, cannot factorial praluction be formed.Therefore, study the rooting method of a kind of Paradox walnut (vlach), and promoted the economic benefit by greatly improving forestry industry, have great importance.
Summary of the invention
The technical problem to be solved in the present invention, be to provide the rooting method in a kind of Paradox walnut crow clarke plantlet in vitro bottle, this rooting method, not by the impact of the factors such as season, for its factorial praluction provides theoretical foundation and technical support, and the seedling that energy is efficient, Quick supplies stable homogeneous, accelerate improved variety popularization speed, and rooting rate can reach 65-75%, booth survival rate 65-85%.
For solving the problems of the technologies described above, the technical solution used in the present invention is:
A rooting method in Paradox walnut crow clarke plantlet in vitro bottle, it comprises the following steps:
(1) choose that plant height is 2-5cm, subculture cycle be 18-23 days, leaf look for green or cyan Paradox walnut plantlet in vitro is as taking root material, excision radical leaves, axillalry bud to base portion reserve the stem section of 1-2 centimetre, obtain taking root material a;
(2) inserted to take root in I medium by the seedling base portion of taking root in material a and carry out light culture, the light culture time is 48-72 hour, and temperature is 20-28 DEG C, obtains material b of taking root;
(3) material b of taking root proceeds to takes root in II medium, and illumination and light culture hocket, and illumination cultivation condition is 13 h light, intensity of illumination 2000-3000 lux, and temperature is 23-27 DEG C; Light culture condition is light culture 11 hours, and temperature is 19-23 DEG C, obtains material c of taking root; Wherein, the nutrient solution of II medium of taking root to be volume ratio be 1:1: matrix;
(4) material c of taking root cultivates 20-24 days, and obtained Paradox walnut crow clarke group trains seedling of taking root.
Limit as one of the present invention, I medium of taking root in described step (2) is: medium based on 1/4-1 part walnut special culture media DKW, add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-6.0.
Limit as another kind of the present invention, the nutrient solution in described step (3) is medium based on 1/4-1 part walnut special culture media DKW, and add 0.01-0.03mg/L indolebutyric acid IBA and 20-50g/L sucrose, pH is 5.5-6.0;
The present invention also has a kind of restriction, and described matrix is vermiculite or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are the vermiculite of 1:1:1: the peat composed of rotten mosses: the one in perlite.
Rooting efficiency corresponding to rooting method provided by the present invention be in view of all technical parameter combinations involved in rooting method after effect, independent employing wherein one or more all can not reach corresponding technique effect, and one or more technical parameters wherein can not be reached corresponding technique effect after simply replacing.
Owing to have employed above-mentioned technical scheme, compared with prior art, acquired technological progress is in the present invention:
The present invention adopts the rooting method in Paradox walnut crow clarke plantlet in vitro bottle, is that Plant Tissue Breeding enters the indispensable step of factorial praluction.Wherein, using the healthy Paradox walnut crow clarke plantlet in vitro to subculture cycle as material of taking root, by it through light culture process, then proceed to root media light culture and dark culture to hocket culture of rootage, after 20-24 days the group training of Planting out of test-tube is taken root seedling, its rooting rate reaches 65-75%, booth survival rate 65-85%.Rooting method in Paradox walnut crow clarke plantlet in vitro bottle provided by the present invention, not by the impact of the factors such as season, for its factorial praluction provides theoretical foundation and technical support, and the seedling that energy is efficient, Quick supplies stable homogeneous, accelerate improved variety popularization speed.
The present invention is applicable to the cultivation of taking root in Paradox walnut crow clarke plantlet in vitro bottle.
The present invention is described in further detail below in conjunction with specific embodiment.
Embodiment
Rooting method in embodiment 1 one kinds of Paradox walnut crow clarke plantlet in vitro bottles
A rooting method in Paradox walnut crow clarke plantlet in vitro bottle, it comprises the following steps:
(1) choose that plant height is 3cm, subculture cycle is 20 days, leaf look is for green or cyan Paradox walnut plantlet in vitro (come from Chinese forest-science academy Xi Shengke and teach laboratory) are as taking root material, excision radical leaves, axillalry bud to base portion reserve the stem section of 1 centimetre, obtain material a of taking root;
(2) inserted to take root in I medium by the seedling base portion of taking root in material a and carry out light culture, the light culture time is 36 hours, and temperature is 24 DEG C, obtains material b of taking root;
Wherein, I medium of taking root is: medium based on 1 part of walnut special culture media DKW, and add 8mg/L indolebutyric acid IBA, 50g/L sucrose and 6g/L agar, pH is 5.5.
(3) material b of taking root proceeds to takes root in II medium, and illumination and light culture hocket, and illumination cultivation condition is 13 h light, intensity of illumination 2500 lux, and temperature is 25 DEG C; Light culture condition is light culture 11 hours, and temperature is 21 DEG C, obtains material c of taking root; Wherein, the nutrient solution of II medium of taking root to be volume ratio be 1:1: matrix;
Wherein, nutrient solution is medium based on 1/4-1 part walnut special culture media DKW, and add 0.01mg/L indolebutyric acid IBA and 30g/L sucrose, pH is 5.5;
Described matrix is vermiculite or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are the vermiculite of 1:1:1: the peat composed of rotten mosses: the one in perlite.
(4) material c of taking root cultivates 24 days, and obtained Paradox walnut crow clarke group trains seedling of taking root, and rooting rate is 75%, and booth growth rate is 85%.
Rooting method in embodiment 2-6 Paradox walnut crow clarke plantlet in vitro bottle
Embodiment 2-6 is respectively the rooting method in a kind of Paradox walnut crow clarke plantlet in vitro bottle, and it is similar to embodiment 1, and difference is only that technical parameter involved in method is different, specifically sees table:
Wherein, take root I medium and the raw material components of taking root in II medium and consumption in embodiment 2-6 are as shown in the table:
Embodiment 7 is taken root the screening of condition
The screening of the dark treatment temperature and time one, in step (2)
| Experimental group | Dark treatment temperature (DEG C) | Dark treatment time (h) | Rooting rate (%) | Booth survival rate (%) |
| 1 | 15 | 36 | 15 | 8 |
| 2 | 18 | 42 | 23 | 40 |
| 3 | 18 | 48 | 62 | 53 |
| 4 | 20 | 42 | 50 | 67 |
| 5 | 20 | 48 | 72 | 83 |
| 6 | 23 | 54 | 70 | 80 |
| 7 | 25 | 63 | 74 | 85 |
| 8 | 28 | 72 | 75 | 81 |
| 9 | 30 | 78 | 68 | 55 |
| 10 | 28 | 78 | 70 | 58 |
| 11 | 25 | 72 | 63 | 72 |
As seen from the above table, best dark treatment temperature is 20-28 DEG C, and the dark treatment time is 48-72h, and rooting rate and booth survival rate, reach 70-75% and 80-85% respectively; But when all non-reach lower limit of dark treatment temperature and time or go beyond the scope upper in limited time, rooting rate and booth survival rate lower; Temperature is wherein in scope when but the time goes beyond the scope or the time, still temperature went beyond the scope in scope, rooting rate and booth survival rate are also starkly lower than the result in scope.Reason is that dark treatment overlong time can cause plant to occur yellow, aging, fallen leaves and generation callus etc., produces harmful effect to adventive root.
Two, to take root the screening of I medium and the raw material components consumption in II medium of taking root
Basal medium adopts DKW medium, adds IBA, sucrose and agar wherein, and the pH value regulated plays very important effect for rooting efficiency.The amount of the minimum use of DKW medium is 1/4 part, if usage amount reduces again, just can not provide necessary nutriment to needing the training seedling of taking root, more than the words of 1 part, its rooting rate and booth growth rate can not be made to be significantly improved, also can cause the increase of production cost, therefore the consumption of the DKW medium of final choice is 1/4-1 part.
As seen from the above table, the optimum proportioning of I medium of taking root is medium based on 1/4-1 part walnut special culture media DKW, and add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-6.0.The lower limit rooting rate and the booth survival rate that exceed provided scope obviously decline, because its nutrient component provided fails to reach the requirement needed for the growth of training seedling; The higher limit rooting rate and the booth survival rate that exceed provided scope significantly do not increase, and can cause the increase of production cost.
As seen from the above table, the optimum proportioning of II medium of taking root is medium based on 1/4-1 part walnut special culture media DKW, and add 0-0.03mg/L indolebutyric acid IBA and 20-50g/L sucrose, pH is 5.5-6.0; The lower limit rooting rate and the booth survival rate that exceed provided scope obviously decline, because its nutrient component provided fails to reach the requirement needed for the growth of training seedling; The higher limit rooting rate and the booth survival rate that exceed provided scope significantly do not increase, and can cause the increase of production cost.
By the screening to above Paradox walnut crow clarke plantlet in vitro root media, finishing screen selects the suitable culture medium prescription that it is taken root and concrete rooting method, and wherein the highest rooting rate reaches 75%, and root system is the normal root system of white, after booth hardening, booth survival rate is up to 85%.Efficiently solve Paradox walnut crow clarke plantlet in vitro to take root difficult problem, for its factorial praluction provides theoretical foundation and technical support, and this technology is not by seasonal effect, and the Paradox walnut crow clarke group training throughout the year can producing stable homogeneous is taken root seedling.After it carries out factorial praluction, be not only the Paradox walnut crow clarke stock seeds that nursery stock markets provides stabilization characteristics of genetics, can be used as the timber forest that reproducting tree species provide a large amount of yet, final for bringing huge economic benefit and ecological benefits in market.
The comparison of embodiment 8 rooting method
Compared for " one-step method is taken root ", " two-step method is taken root " and rooting method provided by the present invention, result is as follows:
" one-step method is taken root " of the prior art refers to that test-tube plantlet is being added with in the medium of growth hormone always and cultivates.
" two-step method is taken root " refers to test-tube plantlet root induction certain hour in a kind of medium, then proceeds to and does not cultivate containing continuing in growth hormone medium.
Because Paradox walnut is taken root more difficult, one-step method is adopted to be difficult to realize its phenomenon of taking root, and walnut is comparatively responsive to auxin substance, the growth hormone of higher concentration is adopted to process for a long time to its basal part of stem, its enlargement can be made and produce callus, this root system produced from callus does not communicate with the conducting system of stem, is unfavorable for the growth of bud seedling, root even can be made to come off; If reduce auxin concentration used, adventive root is not almost had again to occur.
| Rooting method | Rooting rate (%) | Booth growth rate (%) |
| " two-step method " | 25.3-65 | 83 |
| Rooting method of the present invention | 65-75 | 65-85 |
As seen from the above table, using Paradox walnut crow clarke plantlet in vitro as material of taking root, the rooting rate adopting rooting method provided by the present invention to obtain and booth growth rate are the highest.The plantlet in vitro of Paradox walnut crow clarke kind is also described and is not suitable for the general method of prior art.
The above is only preferred embodiment of the present invention, is not restriction the present invention being made to other form, and any those skilled in the art may utilize above-mentioned technology contents to be changed or be modified as the Equivalent embodiments of equivalent variations as enlightenment.But everyly do not depart from technical solution of the present invention content, according to technical spirit of the present invention to the simple modification done by above embodiment, equivalent variations and remodeling, still belong to the protection domain of the claims in the present invention.
Claims (4)
1. the rooting method in Paradox walnut crow clarke plantlet in vitro bottle, is characterized in that it comprises the following steps:
(1) choose that plant height is 2-5cm, subculture cycle be 18-23 days, leaf look for green or cyan Paradox walnut plantlet in vitro is as taking root material, excision radical leaves, axillalry bud to base portion reserve the stem section of 1-2 centimetre, obtain taking root material a;
(2) inserted to take root in I medium by the seedling base portion of taking root in material a and carry out light culture, the light culture time is 48-72 hour, and temperature is 20-28 DEG C, obtains material b of taking root;
(3) material b of taking root proceeds to takes root in II medium, and illumination and light culture hocket, and illumination cultivation condition is 13 h light, intensity of illumination 2000-3000 lux, and temperature is 23-27 DEG C; Light culture condition is light culture 11 hours, and temperature is 19-23 DEG C, obtains material c of taking root; Wherein, the nutrient solution of II medium of taking root to be volume ratio be 1:1: matrix;
(4) material c of taking root cultivates 20-24 days, and obtained Paradox walnut crow clarke group trains seedling of taking root.
2. the rooting method in Paradox walnut crow clarke plantlet in vitro bottle according to claim 1, it is characterized in that I medium of taking root in described step (2) is: medium based on 1/4-1 part walnut special culture media DKW, add 4-16mg/L indolebutyric acid IBA, 30-60g/L sucrose and 4-8g/L agar, pH is 5.5-6.0.
3. the rooting method in Paradox walnut crow clarke plantlet in vitro bottle according to claim 1, it is characterized in that: the nutrient solution in described step (3) is medium based on 1/4-1 part walnut special culture media DKW, add 0.01-0.03mg/L indolebutyric acid IBA and 20-50g/L sucrose, pH is 5.5-6.0.
4. the rooting method in Paradox walnut crow clarke plantlet in vitro bottle according to claim 1, is characterized in that: described matrix is vermiculite or volume ratio is the peat composed of rotten mosses of 1:1: perlite or volume ratio are the vermiculite of 1:1:1: the peat composed of rotten mosses: the one in perlite.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110495395A (en) * | 2019-08-28 | 2019-11-26 | 湖北丰年农业开发有限公司 | A kind of method of Paradox walnut tissue-culturing rapid propagation and industrial seedling rearing |
| CN110741934A (en) * | 2019-11-22 | 2020-02-04 | 塔里木大学 | Method for rooting in early-fruiting walnut tissue culture seedling bottle |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110495395A (en) * | 2019-08-28 | 2019-11-26 | 湖北丰年农业开发有限公司 | A kind of method of Paradox walnut tissue-culturing rapid propagation and industrial seedling rearing |
| CN110741934A (en) * | 2019-11-22 | 2020-02-04 | 塔里木大学 | Method for rooting in early-fruiting walnut tissue culture seedling bottle |
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Effective date of registration: 20230308 Address after: 200333 Room 2403, No. 44, Lane 1817, Zhenbei Road, Putuo District, Shanghai Patentee after: Shanghai Jingyi Biotechnology Co.,Ltd. Address before: Room 2483, Building 10, No. 1630, Yecheng Road, Industrial Zone, Jiading District, Shanghai, 201801 Patentee before: SHANGHAI SHANYI PLANT TECHNOLOGY Co.,Ltd. |