CN104585241A - Preparing and applying method of cutting propagation growth promoting agent of tea tree endogenous herbaspirillum seropedicae - Google Patents
Preparing and applying method of cutting propagation growth promoting agent of tea tree endogenous herbaspirillum seropedicae Download PDFInfo
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- CN104585241A CN104585241A CN201510015515.3A CN201510015515A CN104585241A CN 104585241 A CN104585241 A CN 104585241A CN 201510015515 A CN201510015515 A CN 201510015515A CN 104585241 A CN104585241 A CN 104585241A
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Abstract
The invention provides a preparing and applying method of a cutting propagation growth promoting agent of tea tree endogenous herbaspirillum seropedicae. By using biological characteristics of the tea tree endogenous herbaspirillum seropedicae, a tea tree endogenous herbaspirillum seropedicae culturing solution is prepared by virtue of fermentation of the tea tree endogenous herbaspirillum seropedicae, and is diluted according to a certain ratio and then used for performing soaking treatment on a tea tree cutting wood. Plant hormones secreted by bacteria in the growth promoting agent are used for inducing the cutting wood to form adventitious roots; the bacteria in the growth promoting agent enter the cutting wood to perform endosymbiosis and continuously secret metabolites, so that quick growth of tea seedlings is promoted. The treated and cut cutting wood is developed in root system and high in planting percent, and tea seedlings are darkly green in leaves, and grow vigorously. The technical method provided by the invention is low in cost and simple to operate, is green and safe, and has multiple effects and durable action. The growth promoting agent can be used for cutting propagation of tea trees as commercial crops and can also be used for cutting propagation of other camellia plants.
Description
Technical field
What the present invention relates to is plant propagation growth-promoting agent, particularly a kind of preparation and application method of sward spirillum cottage propagation growth-promoting agent in tea tree.
Background technology
Tealeaves is the important economic crops of China, and 2012 Annual planting areas reach 3,170 ten thousand mu, and annual production 162. 30,000 tons, account for the whole world 37. 8%, cultivated area or output all rank first in the world.Tealeaves is except as except drink, and exploitation effective content of tea has also become new developing direction for medicine, food, chemical industry.Tea tree is cross pollinated plant, and between sexual propagation individuality, economic characters are mixed, and growth differences is large, generally can not keep maternal original characteristic.The advantages such as and tea tree single node cutting propagation technique can keep maternal merit, have reproduction coefficient large, method is easy, and after cuttage root of hair seedling is fast are the main breeding practices of producing upper Extension of Tea Clone Improved Varieties application at present.
For improving tea tree single node cutting reproductive survival rate, during cottage propagation, usually use root-inducing powder.The main component of root-inducing powder is plant hormone substance Nafusaku and the butyric acid potassium of chemosynthesis.Nafusaku promotes the formation of adventive root, the mainly formation of main root, but concentration excessive the suppression takes root; Butyric acid potassium induction crop forms adventive root, the mainly formation of fibrous root, can slowly decompose under high light.Now widely used root-inducing powder is ABT root-inducing powder, has has researched and developed 10 kinds of models, and No. 1-5 is alcohol dissolubility, and No. 6-10 is water-soluble.Tea tree single node cutting breeds No. 1, the ABT and No. 2 that then mainly use alcohol dissolubility, is about to healthy and strong tea tree current-year branch at ABT solution (50mg.kg
-1) soak cuttage after half an hour.Root-inducing powder has the fast advantage of low, the short rooting effect of cost, but effect is single, tea shoot subsequent growth effectively can not be promoted to grow and have the shortcoming of pedo relict risk.
In tea tree, sward spirillum is a kind of endogenetic bacteria strong with Camellia Plants symbiosis selectivity, the document Microbiological Characteristics of sward spirillum " in the two strain tea trees " (Wang Ting etc., microorganism journal 2014 the 4th phase, 2014.04.04, P424-432) do to describe in detail to the screening of sward spirillum in tea tree and Microbiological Characteristics.But the application of sward spirillum in tea tree is not related to.
Summary of the invention
The object of the invention is the preparation and application method proposing sward spirillum cottage propagation growth-promoting agent in a kind of tea tree.With this red stalk of growth-promoting agent process Camellia Plants current-year branch or hard blue or green stalk, cuttage branch can be effectively promoted to take root long seedling.
The present invention realizes like this.Its step is
1, the screening of sward spirillum in tea tree
First rinse disease-free camellia tree sample well with clear water, then use 75% alcohol disinfecting 0.5 hour, sterilize 3 minutes with the mercury solution of percent concentration 1% subsequently.Finally use aseptic water washing 3-5 time.Draw the sterile water 100 μ l of last flushing material, in coating beef-protein medium flat board (3g beef extract, 10g peptone, 5g NaCl, 1.5% agar, add water to 1000ml).37 DEG C cultivate 3 days aseptic drop out existing after, again the tea leaf of disinfecting and tissue are cut marginal portion, interior tissue being cut fritter is positioned in beef extract-peptone culture plate, cultivate after 48 hours for 37 DEG C and obtain endophyte bacterium colony, carry out bacterium colony purifying subsequently and systematic bacteriology is identified, finally obtain sward spirillum in tea tree.
,the cultivation of sward spirillum and the preparation of growth-promoting agent in tea tree
Get sward spirillum bacterial classification in tea tree that-80 ° of C preserve, inoculation 5mL LB liquid nutrient medium (10g tryptone, 5g yeast extract, 10g NaCl, 10 μ g/mL spectinomycins, add water to 1000ml), 150 turns of shakes per minute, 37 ° of C overnight incubation.Bacterium liquid after activation is accessed fresh LB liquid nutrient medium or KB liquid nutrient medium (20 g tryptones, 1.5 g K with 1:100 ratio
2hPO
4, 15 ml glycerine, 1M MgSO
4, 10 μ g/mL spectinomycins, add water to 1000 ml) and middle amplification cultivation, condition of culture is 200 turns per minute of shaking speed, and temperature is 28-37 ° of C.Treat bacterial culture 8-10 hour, OD
600value is for collecting whole inoculums during 1.2-1.5, after adding 1% glycerine, deposit in 4 ° of C for subsequent use or directly use, above-mentioned inoculum is sward spirillum cottage propagation growth-promoting agent in tea tree of the present invention.
The application of described growth-promoting agent:
The process of 1, cuttage skill: the tea tree annual shoot row culture cutting of growth selection stalwartness, the long 7-15 centimetre of cutting, top is truncation above lateral bud, and retain top 1-2 sheet leaf, base portion is truncation below lateral bud.
2, with water by above-mentioned inoculum by 0.5-1:1 dilution proportion and normal temperature (8-38 ° of C) soaks cuttage bar 0.5-3 hour.
3, then carry out cuttage in seedbed, after cuttage, field management method manages routinely.
Mechanism of the present invention is such, and in tea tree, sward spirillum is the endophyte of Theaceae plant, enters in tea tree body, extracellular symbiosis by means of only wound.After the upper and lower truncation of tea cutting bar, in tea tree, sward spirillum enters cutting when soaking by base portion otch.Bacterial metabolism secretion time after cuttage on the one hand by soaking in culture fluid forms adventive root as IAA directly irritates cuttage branch, and the bacterial factor provided by sward spirillum symbiosis in tea tree in cutting body on the other hand promotes tea shoot growth.In tea tree, sward spirillum is natural tea tree endophyte, and disease-free effect, only surely grows usually in older branch or blade, without bacterium in newborn spray tender leaf, meets environmental protection and food safety requirements.
The present invention has unique advantage:
1, the Biological characteristics of sward spirillum in tea tree is taken full advantage of.In tea tree, sward spirillum is the endogenetic bacteria that Camellia Plants selectivity is strong, nuisanceless with tea tree mutualistic symbiosis.It can produce IAA, NH that Promoting plant growth is grown effectively
4 +, addicted to siderophore and polyphenol oxidase.IAA can irritate tea tree Growth of Cells and induced synthesis adventive root, NH
4 +nitrogenous source can be provided for tea tree, for tea tree absorption iron ion and tea tree resistance against diseases can be improved addicted to siderophore, use sward spirillum in tea tree to soak Camellia Plants cuttage bar and can realize the metabolite of bacterial culture and the comprehensive utilization of bacterium itself, cost is low, function is complete, easy to operate.
2, overcome current root-inducing powder function singleness, act on lastingly, effectively can not promote the shortcoming that tea shoot subsequent growth is grown.In tea tree, sward spirillum culture fluid can not only promote the Adventitious root initiation of cuttage branch, and bacterium enter cutting after sustainablely to play a role, promote the quick growth of tea shoot, and be expected to the quality improving tea tree.
Accompanying drawing explanation
Fig. 1 is the embodiment of the present invention 3 tea shoot root, branch growing way picture,
Fig. 2 is the embodiment of the present invention 4 tea shoot root, branch growing way picture,
Wherein a, b, for sward spirillum culture fluid process in tea tree, c, to compare for clear water process.
Embodiment
Below in conjunction with embodiment, the present invention is further described.
Embodiment 1:
Rinse disease-free camellia tree sample well with clear water, then use 75% alcohol disinfecting 0.5 hour, subsequently with 1% mercuric chloride sterilization, 3 minutes.Finally use aseptic water washing 3-5 time.Draw the sterile water 100 μ l of last flushing material, in coating beef-protein medium flat board (3g beef extract, 10g peptone, 5g NaCl, 1.5% agar, add water to 1000ml).37 DEG C cultivate 3 days aseptic drop out existing after, then the tea leaf of disinfecting and tissue are cut marginal portion, interior tissue are cut fritter and be positioned in beef extract-peptone culture plate, cultivate after 48 hours for 37 DEG C and obtain endophyte bacterium colony.Carry out bacterium colony purifying subsequently and systematic bacteriology is identified, finally obtain sward spirillum in tea tree.
Embodiment 2:
Get sward spirillum bacterial classification in tea tree that-80 ° of C preserve, inoculation 5mL LB liquid nutrient medium (10g tryptone, 5g yeast extract, 10g NaCl, 10 μ g/mL spectinomycins, add water to 1000ml), 150 turns of shakes per minute, 37 ° of C overnight incubation.Bacterium liquid after activation is accessed fresh LB liquid nutrient medium or KB liquid nutrient medium (20 g tryptones, 1.5 g K with 1:100 ratio
2hPO
4, 15 ml glycerine, 1M MgSO
4, 10 μ g/mL spectinomycins, add water to 1000 ml) and middle amplification cultivation, condition of culture is 200 turns per minute of shaking speed, and temperature is 28-37 ° of C.Treat bacterial culture 8-10 hour, OD
600value, for collecting whole inoculums during 1.2-1.5, is growth-promoting agent after adding 1% glycerine.
Embodiment 3: the hard blue or green bar cutting in No. 1, mountain, sward spirillum culture fluid process capital 100 3 hours in the tea tree that 0.5:1 is diluted, after cuttage, 100% survives, tea shoot coring after 5 months is flourishing, shoot growing way is vigorous, blade is dark green, is obviously better than control group (see figure 1).
Embodiment 4: the red bar cutting in No. 1, mountain, sward spirillum culture fluid process capital 100 1 hour in the tea tree of dilute 1:1, after cuttage, 100% survives, and the tea shoot coring after 5 months is flourishing, shoot grows fine, blade is dark green, is obviously better than control group (see figure 2).
Claims (2)
1. the preparation of sward spirillum cottage propagation growth-promoting agent in tea tree, is characterized in that the steps include:
1), the screening of sward spirillum in tea tree
Rinse disease-free camellia tree sample well with clear water, then use 75% alcohol disinfecting 0.5 hour, sterilize 3 minutes with the mercury solution of percent concentration 1% subsequently; Finally use aseptic water washing 3-5 time, draw the sterile water 100 μ l of last flushing material, on coating beef-protein medium flat board, 37 DEG C cultivate 3 days aseptic drop out existing after, again the tea leaf of disinfecting and tissue are cut marginal portion, interior tissue being cut fritter is positioned in beef extract-peptone culture plate, cultivates after 48 hours for 28-37 DEG C and obtains endophyte bacterium colony; Carry out bacterium colony purifying subsequently and systematic bacteriology is identified, finally obtain sward spirillum also-80 DEG C of preservations in tea tree;
Described beef-protein medium is 3g beef extract, 10g peptone, 5g NaCl, 1.5% agar, adds water to 1000ml;
2), the cultivation of sward spirillum and the preparation of growth-promoting agent in tea tree
Get sward spirillum bacterial classification in tea tree that-80 ° of C preserve, inoculation 5mL LB liquid nutrient medium, 150 turns of shakes per minute, 37 ° of C overnight incubation; Accessed in fresh LB liquid nutrient medium or KB liquid nutrient medium to amplify with 1:100 ratio by bacterium liquid after activation and cultivate, condition of culture is 200 turns per minute of shaking speed, and temperature is 37 ° of C; Treat bacterial culture 8-10 hour, OD
600value is for collecting whole inoculums during 1.2-1.5, after adding 1% glycerine, deposit in 4 ° of C for subsequent use or directly use, above-mentioned inoculum is sward spirillum in tea tree of the present invention
skewerinsert breeding growth-promoting agent;
Described LB liquid nutrient medium is 10g tryptone, 5g yeast extract, and 10g NaCl, adds water to 1000ml, 10 μ g/mL spectinomycins;
Described KB liquid nutrient medium is 20 g tryptone 1.5 g K
2hPO
4, 15 ml glycerine, 1M MgSO
4, add water to 1000 ml, 10 μ g/mL spectinomycins.
2. the application method of sward spirillum cottage propagation growth-promoting agent in tea tree, is characterized in that the steps include:
1), the process of cuttage skill, the tea tree annual shoot row culture cutting of growth selection stalwartness, the long 7-15 centimetre of cutting, top is truncation above lateral bud, and retain top 1-2 sheet leaf, base portion is truncation below lateral bud;
2), with water above-mentioned inoculum pressed 0.5-1:1 dilution proportion and soak cuttage bar 0.5-3 hour in 4-38 ° of C temperature;
3), then carry out cuttage in seedbed, after cuttage, field management method manages routinely.
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Cited By (5)
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| CN105028485A (en) * | 2015-07-14 | 2015-11-11 | 湖北大学 | Tea tree in-regulating growth promoting agent and application method in expediting sprouts of field tea trees |
| CN106962091A (en) * | 2017-02-15 | 2017-07-21 | 湖北大学 | Sward spirillum promotes the application technology of tealeaves selenium-rich in a kind of utilization tea tree |
| CN109554313A (en) * | 2018-12-25 | 2019-04-02 | 陕西师范大学 | Contain the subspecies H.5-28 cultural method of the bacterial cultures of strain and its application |
| CN116064320A (en) * | 2022-11-24 | 2023-05-05 | 湖北大学 | Tobacco endophytic enterobacter CYLOB for degrading nicotine and application thereof |
| CN116656539A (en) * | 2023-04-20 | 2023-08-29 | 安徽农业大学 | Carpesium V8 and application thereof |
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Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105028485A (en) * | 2015-07-14 | 2015-11-11 | 湖北大学 | Tea tree in-regulating growth promoting agent and application method in expediting sprouts of field tea trees |
| CN105028485B (en) * | 2015-07-14 | 2018-08-31 | 湖北大学 | Regulation and control growth-promoting agent and the application process in crop field tea tree sprouting is expedited the emergence of in a kind of tea tree |
| CN106962091A (en) * | 2017-02-15 | 2017-07-21 | 湖北大学 | Sward spirillum promotes the application technology of tealeaves selenium-rich in a kind of utilization tea tree |
| CN109554313A (en) * | 2018-12-25 | 2019-04-02 | 陕西师范大学 | Contain the subspecies H.5-28 cultural method of the bacterial cultures of strain and its application |
| CN109554313B (en) * | 2018-12-25 | 2021-09-24 | 陕西师范大学 | Method for culturing bacterial culture containing subspecies H.5-28 strain and application thereof |
| CN116064320A (en) * | 2022-11-24 | 2023-05-05 | 湖北大学 | Tobacco endophytic enterobacter CYLOB for degrading nicotine and application thereof |
| CN116064320B (en) * | 2022-11-24 | 2023-11-03 | 湖北大学 | Tobacco endophytic enterobacter CYLOB for degrading nicotine and application thereof |
| CN116656539A (en) * | 2023-04-20 | 2023-08-29 | 安徽农业大学 | Carpesium V8 and application thereof |
| CN116656539B (en) * | 2023-04-20 | 2024-03-22 | 安徽农业大学 | Carpesium V8 and application thereof |
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Application publication date: 20150506 |