CN102749222A - Method and device for collecting root exudates of bulb propagation type plants - Google Patents
Method and device for collecting root exudates of bulb propagation type plants Download PDFInfo
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- CN102749222A CN102749222A CN2012102457559A CN201210245755A CN102749222A CN 102749222 A CN102749222 A CN 102749222A CN 2012102457559 A CN2012102457559 A CN 2012102457559A CN 201210245755 A CN201210245755 A CN 201210245755A CN 102749222 A CN102749222 A CN 102749222A
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Abstract
The invention discloses a method and a device for collecting root exudates of bulb propagation type plants. The method includes 1), breaking dormancy of bulbs; 2), culturing root systems of the bulbs in advance; 3), collecting the root exudates; and 4), roughly separating the root exudates. The device mainly comprises a bulb plant culturing portion, a root exudates collecting portion and a root exudates roughly separating portion. The method is simple and feasible, and the device is long in collecting time and high in efficiency.
Description
Technical field
The invention belongs to the plant cultivation technical field, relate to a kind of collection method and device of secretions from plant roots, be specifically related to a kind of collection method and device of bulb breeding type secretions from plant roots.
Background technology
The collection method of root exudates is varied at present, and the gathering-device corresponding with it also is countless, but do not carry out system divides for the collection method of root exudates.Mainly comprise: solution training collecting method, flooding culture matrix (vermiculite training, sand culture, agar training, earth culture etc.) collecting method, root division device collecting method, make pottery a plastic tube decompression original position collecting method, chromatography filter paper or cellulose membrane location collecting method, soil liquid ST collecting method, isotope labeling combination soil liquid ST collecting method and method that some are derived thereof etc. of continuous apparatus collecting method, porous automatically.Each tool relative merits of these methods, thereby take which kind of collection mode to depend on research object and purposes to a great extent.
Bulb, some spermatophyte, the especially perennial monocotyledonous abnormal stem that is in the dormant stage.Bulb mainly contains two types, and one type is representative with the onion, and the outer film coated with pattern of fleshy leaf is as a means of protection; Another kind of type is shown in typical Liliaceae (Liliaceae) plant, and storage leaf is exposed, and no pattern film covers, and outward appearance is like having the scale of corner angle to constitute by many.Bulb is a kind of of underground abnormal stem.The very cripetura of abnormal stem is plate-like, on it give birth to the scale leaf of plump fleshiness, interior storage is rich nutrient substances and moisture very.Have several kinds of bulbaceous plants good or nutritious, the mankind had important economic value because of its fleshy leaf flavor, for example onion and with it akin verdant, garlic, leek etc.In addition, the root system of green onion garlic class plant can be secreted a kind of mastery rhzomorph, and various bacteria, fungi etc. is had stronger inhibiting effect, and is relatively good to succession crop, is usually used in intercropping or interplanting.Therefore, the research of its root exudates is also more and more obtained paying attention to, it is vital producing a kind of specific root exudates collection method and device to this type of plant thus.
Summary of the invention
The objective of the invention is to overcome above-mentioned technological deficiency, a kind of collection method and device of bulb breeding type secretions from plant roots is provided.This method is simple, and this device acquisition time is long and efficient is high.Its technical scheme is:
A kind of collection method of bulb breeding type secretions from plant roots may further comprise the steps:
1) abolishes the bulb dormancy
The selection growing way is even, the bulb of no disease and pest, and its low temperature environment that is put in 0-15 ℃ is handled 0-30 days to break its physiological dormancy;
2) the bulb root system is cultivated in advance
Bulb was at first soaked 20-40 minute with the 0.2%-1.0% liquor potassic permanganate; Be planted in then in the matrix (perlite) of the bacterium of going out, treat the bulb root growth, take out to 3-8 centimetre; Clean, transfer to bulb class secretions from plant roots and collect and the roughing out device;
3) root exudates is collected
The bulb of having taken root is placed in the water planting groove that distilled water is housed, is fixed with cystosepiment in the water planting groove, before cystosepiment is fixing; The hole of suitable bulb size on the brill, above bulb was mounted to, root system was dipped in the distilled water; At room temperature with the continuous aerobic culture of inflator pump; The drainage tube of inflator pump passes cystosepiment and reaches in the distilled water, regularly derives the nutrient solution that contains root exudates, and injects the dark 3-5 of bottom land liquid centimetre distilled water to the water planting groove;
4) root exudates roughing out
Through the coarse filtration and the roughing out of filter cotton and activated charcoal realization root exudates, collect the activated charcoal of absorption root exudates.
Be 30-45 days between culture period in the step 3).
Nutrient solution of collection in every 5-10 days between culture period in the step 3).
A kind of gathering-device of bulb breeding type secretions from plant roots comprises that mainly the bulb plant is cultivated, root exudates is collected and root exudates roughing out three parts,
Said bulb plant is cultivated part and is connected in sequence by water planting groove 1, cystosepiment 2, feed liquor device 3 and inflator pump 4;
Said root exudates is collected part and is connected in sequence by drainage tube 5 and valve 6;
Said root exudates roughing out part is connected in sequence by filtrator 7, separating column 9, wastewater trough 10.
Further preferred, fill filter cotton in the said filtrator 7.
Further, fill activated charcoal 8 in the said separating column 9.
Compared with prior art, beneficial effect of the present invention is:
Technical scheme specific aim of the present invention is more intense, need not add any nutritional labeling between culture period, and the collection cycle was 30-45 days.Through charcoal absorption, realized the roughing out of root exudates simultaneously, the extraction through specific organic solvent can directly be used for follow-up research, the for example biological test of root exudates, bacteriostatic test and material evaluation etc.
Description of drawings
Fig. 1 is the gathering-device structural representation of bulb breeding type secretions from plant roots of the present invention;
Fig. 2 is the inhibition design sketch remarks of variable concentrations garlic root secretion to the phytophthora blight of pepper mycelial growth: T0: contrast T1: (0.02%w/v) 25% T2: (0.04%w/v) 50% T3: (0.06%w/v) 75% T4: (0.08%w/v) 100%;
The lily root exudates sense combined action intensity contrast design sketch that Fig. 3 collects for different phase.
Embodiment
Below in conjunction with accompanying drawing and embodiment technical scheme of the present invention is done explanation in further detail.
With reference to Fig. 1; A kind of gathering-device of bulb breeding type secretions from plant roots; Comprise that mainly the bulb plant is cultivated, root exudates is collected and root exudates roughing out three parts, said bulb plant is cultivated part and is connected in sequence by water planting groove 1, cystosepiment 2, feed liquor device 3 and inflator pump 4; Said root exudates is collected part and is connected in sequence by drainage tube 5 and valve 6; Said root exudates roughing out part is connected in sequence by filtrator 7, separating column 9, wastewater trough 10.Further preferred, fill filter cotton in the said filtrator 7.Further, fill activated charcoal 8 in the said separating column 9.
Through the drainage tube 5 on water planting groove right side, open valve 6 when collecting nutrient solution, liquid just flows out through drainage tube.Effluent at first carries out coarse filtration through the filter cotton 7 in the drainage tube, and the filter cotton aperture can be adjusted according to the research needs voluntarily, and liquid flows into and is equipped with in the separating column 9 of activated charcoal 8 then, and charcoal absorption root exudates, waste liquid are discharged to wastewater trough 10.So just, collected the roughing out thing of bulb root exudates.According to the follow-up study needs, adopt specific organic solvent to extract predetermined substance.
Adopt seed germination experiment that the allelopathy of two garlic cultivar root exudates is carried out biologicall test.
The test concrete steps are: the garlic root of getting variable concentrations is collected liquid 9mL and is added in the plastic containers that are covered with 2 layers of qualitative filter paper (diameter is 12cm) and carry out seed germination experiment; The lactuca sativa seeds of 100 uniformities of every ware sowing; With distilled water is contrast; Each handles 3 repetitions, under 22 ± 1 ℃, 4000Lx illumination, cultivates, and replenishes every day and collects liquid 2mL.Investigate its germinative number every day, calculate germination percentage, germination index at last, measure the length of overground part and root behind the 10d, weighing seedling fresh weight.The mensuration of fresh weight is a repetition with 10 seedlings, and the germination standard is the half the of seed length.
The germinative number Dt-of the Gt-seed soaking back t day number of days that germinates accordingly in the formula
The germination percentage calculation date is: 7d; The germination index calculation date is: 3d.
Table 1 garlic root secretion is to the influence of lactuca sativa seeds germination and growth of seedling
Can find out by table 1, except that germination index along with the rising of garlic root secretion concentration reducing, the Changing Pattern of other each indexs all demonstrates the trend that low concentration promotes that high concentration suppresses.
(1) preparation method of flat-plate bacterial colony
It is 10 that the phytophthora blight of pepper mycelia that activation is good uses sterilized water to be mixed with spore concentration
4~10
5Individual mL
-1Bacteria suspension, the bacteria suspension of drawing 100 μ L are wipeed off with aseptic glass spatula in PDA nutrient culture media (containing potato 200g, agar 18g, sucrose 20g among the 1000mL) flat board.28 ℃ of dark conditions are cultivated 3d down, treat to be used to prepare the bacterium colony disk after bacterium colony covers with flat board.
(2) contain the preparation of garlic root secretion nutrient culture media
The activated charcoal of collecting that contains allelochemical is used the acetone wash-out, ethyl acetate extraction, 40 ℃ of rotation evaporates to dryness.The garlic root secretion dry that obtains is made into 0.02%w/v (T1) by volume with sterilized water, 0.04%w/v (T2), four concentration gradients of 0.06%w/v (T3) and 0.08w/v (T4) are contrast with the sterilized water.Under the aseptic condition, after the garlic root secretion of variable concentrations filtered with sterilizing filter (miillpore filter, aperture 0.45um), get 1ml and sterilized 15mlPDA nutrient culture media (containing potato 300g, agar 25g, sucrose 30g among the 1000mL) and mix.
(3) bacteriostatic activity is measured
Adopt growth rate method to measure bacteriostatic activity.Beat from the flat board that covers with bacterium colony with the card punch of diameter 7mm and to get the bacterium colony disk, place on the PDA plating medium that contains variable concentrations garlic root secretion for preparing in advance, 3 in every ware, every processing 3 wares are contrast (CK) with the sterile aqueous media.Place under 28 ℃ of dark conditions and cultivate, measure bacterium colony expansion diameter behind the 3d, and calculate percentage mycelial inhibition.
Bacteriostasis rate (%)=(contrast ware colony diameter-processing ware colony diameter)/contrast ware colony diameter * 100%
Can find out that by Fig. 2 the garlic root secretion of collecting through device of the present invention has significant inhibitory effect to the growth of phytophthora blight of pepper mycelia, and strengthens with the increase of root exudates concentration.
The lily root exudates sense combined action intensity contrast that embodiment 3 different phases are collected
Choose in the liquor potassic permanganate that 100 recipient plant seeds are immersed in concentration 3g/L behind the sterilization 10min; With distilled water flushing 2-3 time, be sowed in the double dish that is covered with 2 metafiltration paper, accurately draw the 8mL lily root exudates that the different water planting stages obtain during beginning; Add in the double dish; Later every 2d adds 2mL, is treated to contrast with distilled water, and each handles repetition 3 times.Under 25 ℃ of temperature, dark condition, cultivate, investigate the germination number every day, calculate percentage of seedgermination behind the 10d, and measure seedling root length, height of seedling, root fresh weight, seedling fresh weight.
Changing sense effect Index for Calculation formula is: and RI=1-C/T (as T >=C), RI=T/C-1 (when T<C).In the formula: RI>0 shows facilitation, and RI<0 shows inhibiting effect, and T representes processing costs, and C representes control value.Order of magnitude is represented effect strong and weak (Williamson 1988).
Can find out that by Fig. 3 the lily root exudates that different phase is collected is different to the allelopathy intensity of isoacceptor not.
The above; Be merely the preferable embodiment of the present invention; Protection scope of the present invention is not limited thereto; Any technician who is familiar with the present technique field is in the technical scope that the present invention discloses, and the simple change of the technical scheme that obtains or equivalence replacement all fall in protection scope of the present invention with may be obvious that.
Claims (6)
1. the collection method of a bulb breeding type secretions from plant roots is characterized in that, may further comprise the steps:
1) abolishes the bulb dormancy
The selection growing way is even, the bulb of no disease and pest, and its low temperature environment that is put in 0-15 ℃ is handled 0-30 days to break its physiological dormancy;
2) the bulb root system is cultivated in advance
Bulb was at first soaked 20-40 minute with the 0.2%-1.0% liquor potassic permanganate, be planted in then in the matrix perlite of the bacterium of going out, treat the bulb root growth, take out, clean, transfer to bulb class secretions from plant roots and collect and the roughing out device to 3-8 centimetre;
3) root exudates is collected
The bulb of having taken root is placed in the water planting groove that distilled water is housed, is fixed with cystosepiment in the water planting groove, before cystosepiment is fixing; The hole of suitable bulb size on the brill, above bulb was mounted to, root system was dipped in the distilled water; At room temperature with the continuous aerobic culture of inflator pump; The drainage tube of inflator pump passes cystosepiment and reaches in the distilled water, regularly derives the nutrient solution that contains root exudates, and injects the dark 3-5 of bottom land liquid centimetre distilled water to the water planting groove;
4) root exudates roughing out
Through the coarse filtration and the roughing out of filter cotton and activated charcoal realization root exudates, collect the activated charcoal of absorption root exudates.
2. the collection method of bulb breeding type secretions from plant roots according to claim 1 is characterized in that, is 30-45 days between culture period in the step 3).
3. the collection method of bulb breeding type secretions from plant roots according to claim 1 is characterized in that, nutrient solution of collection in every 5-10 days between culture period in the step 3).
4. the gathering-device of a bulb breeding type secretions from plant roots is characterized in that, comprises that mainly the bulb plant is cultivated, root exudates is collected and root exudates roughing out three parts,
Said bulb plant is cultivated part and is connected in sequence by water planting groove (1), cystosepiment (2), feed liquor device (3) and inflator pump (4);
Said root exudates is collected part and is connected in sequence by drainage tube (5) and valve (6);
Said root exudates roughing out part is connected in sequence by filtrator (7), separating column (9), wastewater trough (10).
5. the gathering-device of bulb breeding type secretions from plant roots according to claim 4 is characterized in that, fills filter cotton in the said filtrator (7).
6. the gathering-device of bulb breeding type secretions from plant roots according to claim 4 is characterized in that, fills activated charcoal (8) in the said separating column (9).
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Cited By (6)
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CN103411793A (en) * | 2013-07-18 | 2013-11-27 | 吉林农业大学 | Apparatus for collecting root secretions through aerosol culture process, and collecting method thereof |
CN103759978A (en) * | 2014-02-10 | 2014-04-30 | 金陵科技学院 | Multi-combined device for sterile collection of plant root exudates and application thereof |
CN103843601A (en) * | 2012-11-30 | 2014-06-11 | 江苏大学 | Circular combined type plant root collecting device |
CN109717213A (en) * | 2019-01-28 | 2019-05-07 | 广西大学 | A kind of disease-resistant tomato stock root exudates collection liquid and its suppressing method to ralstonia solanacearum |
CN109717212A (en) * | 2019-01-28 | 2019-05-07 | 广西大学 | A kind of method that disease-resistant stock root exudates is collected and extracted |
CN114112480A (en) * | 2021-12-17 | 2022-03-01 | 贵州省生物研究所 | Device and method for collecting pepper rhizosphere secretion |
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CN101294874A (en) * | 2008-06-12 | 2008-10-29 | 中国科学院东北地理与农业生态研究所 | Crop root secretion collecting device and method thereof |
CN202676509U (en) * | 2012-07-17 | 2013-01-16 | 西北农林科技大学 | Device for collecting root exudates of bulbus propagation plants |
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CN101294874A (en) * | 2008-06-12 | 2008-10-29 | 中国科学院东北地理与农业生态研究所 | Crop root secretion collecting device and method thereof |
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Cited By (8)
Publication number | Priority date | Publication date | Assignee | Title |
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CN103843601A (en) * | 2012-11-30 | 2014-06-11 | 江苏大学 | Circular combined type plant root collecting device |
CN103843601B (en) * | 2012-11-30 | 2015-11-18 | 江苏大学 | Circulation combined type root exudates harvester |
CN103411793A (en) * | 2013-07-18 | 2013-11-27 | 吉林农业大学 | Apparatus for collecting root secretions through aerosol culture process, and collecting method thereof |
CN103411793B (en) * | 2013-07-18 | 2015-07-22 | 吉林农业大学 | Apparatus for collecting root secretions through aerosol culture process, and collecting method thereof |
CN103759978A (en) * | 2014-02-10 | 2014-04-30 | 金陵科技学院 | Multi-combined device for sterile collection of plant root exudates and application thereof |
CN109717213A (en) * | 2019-01-28 | 2019-05-07 | 广西大学 | A kind of disease-resistant tomato stock root exudates collection liquid and its suppressing method to ralstonia solanacearum |
CN109717212A (en) * | 2019-01-28 | 2019-05-07 | 广西大学 | A kind of method that disease-resistant stock root exudates is collected and extracted |
CN114112480A (en) * | 2021-12-17 | 2022-03-01 | 贵州省生物研究所 | Device and method for collecting pepper rhizosphere secretion |
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Application publication date: 20121024 |