CN101492647A - Method for reinforcing salt endurance of vegetable crop - Google Patents
Method for reinforcing salt endurance of vegetable crop Download PDFInfo
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- CN101492647A CN101492647A CNA2008101625527A CN200810162552A CN101492647A CN 101492647 A CN101492647 A CN 101492647A CN A2008101625527 A CNA2008101625527 A CN A2008101625527A CN 200810162552 A CN200810162552 A CN 200810162552A CN 101492647 A CN101492647 A CN 101492647A
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Abstract
The invention relates to a method for enhancing the salt endurance of vegetable crops and belongs to the technical field of edaphon. In the invention, a pseudomonad strain DW1 CGTCC No.2729 with high 1-amino-cyclopropane 1-carboxylic acid deaminase activity is first screened and obtained from coastal soils; further more, a method for enhancing the salt endurance of vegetable crops is proposed; the method comprises the steps as follows: the preparation of a DF culture medium; the activating culture of the strains; the fermentation culture of the strains; the screening of salt-resistant vegetable variety, and the construction of vegetable variety-microbic system. The method can remarkably improve the fresh weight of the vegetable crops under a salt concentration of 0.1 percent and 0.2 percent as well as the content of Ca<2+>and k<+> ions, reduce the content of proline and Na<+> ion, simultaneously increase the antioxidase activity of seedlings, conduce to the plant growth under a salt stress condition and improve the productivity. The invention can be popularized and applied in improving and using seashore saline soil.
Description
Technical field
The present invention relates to the soil microorganisms technical field, particularly relate to by separation, the bacterial strain that screens and the vegetable variety that better salt tolerance is arranged with 1-amino-cyclopropane 1-carboxylic acid (ACC) deaminase active, by making up the associating approach of " plant-microorganism ", to realize alleviating the method for tidal land soil salt to the vegetable crop detrimental effect.
Background technology
Agricultural is the Primary Industry that the mankind depend on for existence.Along with Increase of population, also increasing to the demand of agricultural, and the salinification of soil has limited Agricultural Development, becomes a global resource and ecological problem.According to statistics, irrigateing land of the arable land in the whole world 20% and nearly half all is subjected in various degree salt stress.With regard to China, the whole nation has above 1 * 10
9Hm
2Various salt marsh soil, wherein modern saline soil about 0.373 * 10
9Hm
2, remaining saline soil about 0.446 * 10
9Hm
2, other potential saline soil about 0.173 * 10
9Hm
2In the irrigation district, soil salinization area is just with annual 10
5Hm
2Speed development.The marine solonchak of region following the line of the sea, Zhejiang is typical saline soil.The improvement utilization of solonchak is the important measures that solve the problems such as population, grain, resource and environment that face.
Salt stress can influence phytomorph and grow, and reduces photosynthesis of plant, impels the vegetable cell dehydration, destroys membrane structure, influences the plant materials intracellular metabolite, changes plant lipid and protein component, and then influences the growth of plant.Be to improve the salt tolerance of plant, traditional means is mainly concentrated trench digging hide salt, the ditching draining desalinization of soil by flooding or leaching, fill plantation etc., but along with the wretched insufficiency of Freshwater resources, forces people to consider to utilize plant improvement to utilize solonchak.Along with development of biology, make plant fully adapt to the salt marsh environment by biological approach, to improve the productivity of plant on salt-affected soil, be the new direction that improves the saline soil productivity both at home and abroad in recent years, as passing through molecular breeding or seed treatment to improve the salt tolerance of plant.
Plant rhizosphere microbe is of a great variety and active, constitutes distinctive rhizosphere soil microorganism fauna and be to improve plant to one of effective means of bad edatope resistance.Wherein, plant growth-promoting rhizosphere bacterium (PGPR) is that a class can be by synthetic certain compound (as tethelin) for plant utilization, or the promotion plant is to the absorption of nutritive substance, or by biological control to pathogenic micro-organism, alleviate or suppressed deleterious rhizosphere microorganism, thereby promote the beneficial microorganism of plant-growth indirectly, its application is very extensive.In present research report, discovery achromobacters such as Shimon.Mayak (Achromobacter piechaudii) can promote growth (the Plant growth-promoting bacteria confer resistance intomato plants to salt stress of tomato under salt stress, Plant Physiology and Biochemistry, 2004,42:565-572); In the research of Azospirillum (Azospirillum lipoferum), discoveries such as Bacilio.M this Pseudomonas under salt stress can be colonizated in wheat root and promote the growth of wheat (Mitigation of salt stress inwheat seedling by a gfp-tagged Azospirillum lipoferum, Biological Fertilization andSoils, 2004,40:188-193).All studies show that helps alleviating the infringement that plant is subjected to salt stress by making up suitable rhizospheric microorganism.But do not see as yet so far and utilize pseudomonas and plant to make up " plant-microorganism " system, thereby improve the report of vegetable crop at strand salinate fields upgrowth situation.
The present invention is colonizated in the vegetable crop rhizosphere by the rhizospheric microorganism that utilization has 1-amino-cyclopropane 1-carboxylic acid (ACC) deaminase active, reduces ethylene content in the plant materials, thereby alleviates the infringement of salt stress to plant materials, improves the salt tolerance of plant.Combine with the screening of traditional plant salt tolerant kind simultaneously, " plant-microorganism " system of structure, alleviate the influence that vegetable crop is coated with the ground salinity, improve plant at tidal land upgrowth situation on the ground, optimize the upgrowth situation of vegetable crop on tidal land soil, to improve the productivity of plant on tidal land.
Summary of the invention
The present invention seeks to, the salt stress infringement at tidal land ground salinity causes plant provides a kind of and can strengthen the vegetable crop salt tolerance, improves the method for vegetable crop at strand salinate fields upgrowth situation.
Ultimate principle of the present invention is: by the screening to microorganism active bacterium and vegetable variety, obtain having the microorganism active bacterial strain and the stronger vegetable variety of salt tolerance of high 1-amino-cyclopropane 1-carboxylic acid deaminase active respectively; Then the vegetable variety seed is soaked in microorganism active bacterium bacterium liquid, dressing, constitute distinctive rhizosphere soil microorganism fauna; At last this seed is planted on highly-saline tidal land, by utilizing the precursor ACC of microorganisms acc deaminase catalyzed ethylene, thus the ethylene levels when influencing plant-growth, to obtain higher yield and quality.
Objects of the present invention are achieved through the following technical solutions:
A kind of screening and evaluation with 1-amino-cyclopropane 1-carboxylic acid deaminase active bacterium:
The enrichment culture of a, bacterial classification: will be suspended in from the soil sample of Shangyu, Zhejiang tidal land collection the PAF substratum, in 28-35 ℃ of shaking table 200rmin
-1Shaking culture 24h gets its suspension and is forwarded in the DF substratum, cultivates 24h under same processing condition, promptly obtains enrichment culture liquid;
B, substratum preparation and strain separating purifying thereof: get step a enrichment culture liquid and transfer, under same processing condition, cultivate 24-48h to the bacterium logarithmic phase in being in the DFa substratum of only nitrogen source with 1-amino-cyclopropane 1-carboxylic acid; Get bacteria suspension and on the separation and purification substratum, be coated with flat board, after cultivating 72h under the same processing condition, the single bacterium colony that picking is dissimilar, continuous purification is 5 times on substratum, obtains the pure culture bacterium;
The screening of c, bacterial classification: to the bacterium of Dfa inoculation of medium through step b purifying, each bacterial strain repeats for 3 times; Place 28 ℃-35 ℃ thermostat container to cultivate 24-72h in culture dish, measure the acc deaminase activity of each bacterial strain, therefrom select the highest bacterial strain of acc deaminase activity, called after DW1;
The evaluation of d, bacterial classification: the Physiology and biochemistry by " common bacteria system identification handbook " and " uncle's Jie Shi Bacteria Identification handbook (the 8th edition) " is identified, the form of bacterial strain DW1 is shaft-like, no brood cell, amphitrichous, Gram-negative, can produce fluorochrome, on the LB flat board, be milky white translucent, neat in edge; Identify that through Physiology and biochemistry this strain microorganism has the acc deaminase activity, its enzyme work is up to 0.023U/mg; Binding molecule is identified, is confirmed that this bacterial strain is pseudomonas (Pseudomonas sp.).
The preservation of biological sample material: this pseudomonas (Pseudomonas sp.) bacterial strain DW1 has been deposited on October 29th, 2008 that " China Committee for Culture Collection of Microorganisms's common micro-organisms " center ", deposit number are CGMCC No.2729.
A kind of method that strengthens the vegetable crop salt tolerance, the concrete steps of this method are:
(1) culture medium preparation: the DF substratum, every L contains KH
2PO
44.0g, Na
2HPO
46.0g, MgSO
47H
2O 0.2g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, (NH
4)
2SO
42.0g, pH 7.2-7.4; The DFa substratum joins ACC and does not contain (NH
4)
2SO
4The DF substratum in, its final concentration is 〉=2.0mmolL
-1, after the sterilization, standby;
(2) activation culture of bacterial classification: with pseudomonas (Pseudomonas sp.) bacterial strain DW1, CGMCCNo.2729 is inoculated in the DFa substratum and activates, and treats that bacterium grows to logarithmic phase, and is standby;
(3) fermentation culture of bacterial classification: 100 μ l insert in step (1) the DF substratum with step (2) activation bacteria suspension, cultivate 24-48h down at 25 ℃-37 ℃, to thalline logarithmic growth after date, carry out centrifugation, obtain thalline, and are standby;
(4) screening of salt tolerant vegetable variety: a, to the comparison of various vegetable crop seed germination rates, filter out the vegetable variety that higher germination rate is arranged under condition of salt stress; B, the vegetable variety higher to the seed germination rate that filters out among the step a carry out the experiment of growth of seedling salt tolerance, measure by plant growth condition, therefrom filter out the vegetable variety of salt tolerant;
(5) structure of vegetable variety-microflora: it is A that step (3) fermentation thalline is mixed with concentration
600〉=0.1 bacteria suspension; After the salt tolerant vegetable variety seed disinfection processing that step (4) is filtered out, sterilized water are cleaned, in bacteria suspension, soak, dressing, make the seed-coat bacteria containing amount reach 3.0 * 10
7-6.0 * 10
7CFU/, can sow after draining, plant.
Beneficial effect of the present invention:
The one, the present invention separates the new bacterial strain DW1 with higher 1-amino-cyclopropane 1-carboxylic acid deaminase active from tidal land soil; this bacterial strain can be that nitrogenous source is grown with 1-amino-cyclopropane 1-carboxylic acid; its enzyme work is up to 0.023U/mg; can be by reducing 1-amino-cyclopropane 1-carboxylic acid content in the plant materials; and then the ethylene content that causes by salt stress in the minimizing plant materials; thereby protective plant avoids the murder by poisoning of excessive ethene, guarantees the growth of plant at salinate fields.
The 2nd, the present invention by salt tolerance relatively filters out from a large amount of existing vegetable varieties and draws eggplant No. one and the vegetable variety of assorted two salt tolerants of tomato in Zhejiang, and its salt tolerant threshold value all reaches 2gNaCl/kg soil (seeing Table 1).
The 3rd, by the structure of plant-microorganism system, can significantly increase fresh weight and the Ca of eggplant under 0.1% and 0.2% salt concn
2+And k
+Ionic content reduces proline(Pro) and Na
+Ion content, the while can increase the fresh weight and the activities of antioxidant enzymes of tomato seedling, helps the growth of vegetable crop under condition of salt stress, improves the productivity of vegetables on tidal land, and then is used to coastal saline soil improvement (seeing Table 2,3).
Embodiment
The present invention is described in further detail below by embodiment, but should be appreciated that the present invention is not limited by these contents.
ACC: U.S. sigma company produces, purity 〉=99.9%.
Embodiment 1:(has the screening of 1-amino-cyclopropane 1-carboxylic acid deaminase active bacterial strain)
Carry out according to the following steps:
(1) enrichment culture of bacterial classification: Shangyu, Zhejiang tidal land soil sample is suspended in (every L contains the 10g peptone, 10g casein hydrolysate, 1.5g MgSO in the PAF substratum
4, 1.5g K
2HPO
4), in 28 ℃ of-35 ℃ of shaking table 200rmin
-1Shaking culture 24h gets suspension and is forwarded to that (every L contains 4.0gKH in the DF substratum
2PO
4, 6.0g Na
2HPO
4, 0.2g MgSO
47H
2O, 2.0g glucose, 2.0g gluconic acid, 2.0g citric acid, 2.0g (NH
4)
2SO
4, pH7.2-7.4), cultivate 24h with condition, promptly obtain enrichment culture liquid;
(2) culture medium prescription and strain separating purifying: get enrichment culture liquid transfer in the DFa substratum that with 1-amino-cyclopropane 1-carboxylic acid is only nitrogen source (ACC be dissolved in ultrapure water after suction filtration sterilization, be added to and do not contain (NH
4)
2SO
4And in advance in Mie Jun the DF salt culture medium, ACC final concentration 〉=2.0mmolL
-1) in, cultivate 24-48h to the bacterium logarithmic phase with condition; Get bacteria suspension and on purifying substratum (add agar 20g/L), be coated with flat board in the Dfa substratum, culture dish is placed be inverted in the thermostat container of the same temperature and cultivate 72h, the single bacterium colony that picking is dissimilar, continuous purification is 5 times on substratum, obtains pure culture;
(3) screening of bacterial classification and enzyme activity determination: under aseptic condition, the Dfa purifying substratum of sterilization is poured in the culture dish under 50 ℃ of temperature, after cooling to the bacterium of inoculation of medium through step (2) purifying, each bacterial strain repeats for 3 times: place the thermostat container of the same temperature to cultivate 24-72h in culture dish, (cell suspension of getting 200 μ l is in the 1.5ml centrifuge tube for the acc deaminase activity of mensuration bacterial strain, add 20 μ l 0.5MACC, the vibration mixing; Behind 30 ℃ of insulation 15min, add 1ml 0.56M HCl, vibration mixing, the centrifugal 5min of 16000G under the room temperature; Get the 1ml supernatant liquor, add 800 μ l 0.56M HCl, behind the mixing, add 300 μ l 2,4 dinitrophenyl hydrazines, be incubated 30min in 30 ℃ behind the mixing; With 2.0ml 2M NaOH colour developing, measure absorbance value in the 540nm place then; Calculate the concentration of α-batanone acid by typical curve, be converted into the enzyme unit that lives; 1 enzyme unit definition alive: under these conditions, acc deaminase catalysis ACC, per minute generate the activity of 1 μ mol α-batanone acid), select the highest bacterial strain of acc deaminase activity, promptly obtain required microorganism active bacterium, called after bacterial strain DW1.
Following examples bacterial strain uses therefor is the bacterial strain DW1 that present embodiment screens, and the cell extract of 200 μ l DW1 reacts 30min in the enzyme reaction system, and A540 is 0.153.According to typical curve, be equivalent to per minute and produce 8.98 * 10
-4μ mol α-batanone acid.Cell extract weight in wet base 0.195 μ g/ μ l, the enzyme of its cell extract live and are 0.023U/mg.
Embodiment 2:(has the evaluation of 1-amino-cyclopropane 1-carboxylic acid deaminase active bacterial strain DW1)
The bacterial strain DW1 that is screened by embodiment 1 is by " uncle's Jie Shi Bacteria Identification handbook (the 8th edition) ", and " common bacteria system identification handbook " carries out the physio-biochemical characteristics evaluation, and press molecular cloning three and extract total DNA, utilize primer: BSF8/20,5 '-AGAGTTTGATCCTGGCTCAG-3 ' (Escherichia coli correspondence position is 8-27); Reverse primer BSR1541/20, the 16S rDNA gene of 5 '-AAGGAGGTGATCCAGCCGCA-3 ' (the E.coli correspondence position is 1541-1522) amplification bacterial strain, entrust Invitrogen company to this bacterium 16S rDNA amplification and order-checking, after obtaining the 16S rDNA sequence of this bacterial strain, on the NCBI website, retrieve the 16S rRNA gene order of relevant bacterial strain among the GenBank, and carry out the homology comparison with BLAST.Evaluation based on aspects such as form, physiological and biochemical property and 16SrDNA sequences, DW1 is accredited as pseudomonas with bacterial strain, intend called after pseudomonas (Pseudomonas sp.) DW1, this bacterial strain is deposited in Chinese common micro-organisms preservation center on October 29th, 2008, and deposit number is CGMCC No.2729.
The screening of embodiment 3:(salt tolerant vegetable variety under condition of salt stress)
Carry out according to the following steps:
(1) seed germination experiment: in diameter 9cm culture dish, carry out NaCl concentration and be respectively: 0 (CK), 0.1%, 0.2%, 0.3% seed germination culture experiment, all concentration are all established 3 repetitions; 30 seeds of each culture dish sowing, the NaCl solution 10ml of a certain concentration of disposable adding during each is handled, 28 ℃ of culture temperature write down the chitting piece number and calculate percentage of germination in the 7d that germinates; Percentage of germination (%)=7d chitting piece number/sowing seed number * 100%, it is long to measure bud simultaneously;
(2) potted plant experiment: select the high kind of rate of emergence further to carry out potted plant experiment, the test alms bowl is the polypots of high 12cm, diameter 10cm, every alms bowl dress soil (Hangzhou Wan marine solonchak) 0.5kg, and execute urea 0.216g, potassium primary phosphate 0.096g, vitriolate of tartar 0.124g; Sleeving plastic bag (preventing that salinity from losing with current) in the alms bowl; Salinity (NaCl) is handled 6 levels of establishing, and promptly 0.57,1.0,1.5,2.0,2.5 and 3.0gkg
-1,, pour into for examination soil after analytical pure NaCl dissolves with suitable quantity of water for making soil body salinity even, and stir, every alms bowl is broadcast 10 seeds, stays 4 strains after emerging, and weighs and waters, keep 75% of field capacity, place SANYO GS (SANYO) MLR-350H type illumination box to cultivate (illumination every day 14h, 26 ± 0.5 ℃ of day temperatures, 20 ± 0.5 ℃ of evenings), get the overground part oven dry behind the 60d, weigh; Random alignment is adopted in test, repeats 4 times (seeing Table 1).
The result shows, wherein draw eggplant No. one and the assorted tomato salt-tolerant in Zhejiang good, its salt tolerant limit all reaches 2gNaCl/kg soil.
The The selection result of table 1 salt tolerant vegetable variety under condition of salt stress
Precocious five Zhejiang Bai Jiuda rascal Japan are big
Salt concn is drawn the assorted kind Zhejiang powder designation Da Bai Da Bai broad bean broad bean in eggplant Zhejiang, a Hangzhoupro of eggplant
(g/kg soil) is (g) eggplant (g) eggplant (g) dish (g) dish (g) (g) (g) (g)
ck 1.13 1.07 4.67 3.4 1.18 0.78 2.44 1.38
1 1.35 1.1 3.35 3.32 1.2 0.88 1.6 1.21
1.5 0.98 0.35 2.4 2.28 1.19 0.92 1.22 1.1
2 0.74 0.27 2 1.74 0.85 0.78 0.92 0.7
Embodiment 4:(draws the structure and the potted plant experiment of No. one-microflora of eggplant)
Carry out according to the following steps:
(1) culture medium preparation: the DF substratum, every L contains KH
2PO
44.0g, Na
2HPO
46.0g, MgSO
47H
2O 0.2g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, (NH
4)
2SO
42.0g, pH 7.2-7.4; The DFa substratum joins ACC and does not contain (NH
4)
2SO
4The DF substratum in, its final concentration is 〉=2.0mmolL
-1, after the sterilization, standby; After the sterilization, standby;
(2) activation culture of bacterial classification: with pseudomonas (Pseudomonas sp.) bacterial strain DW1, CGMCCNo.2729 is inoculated in the DFa substratum and activates, and treats that bacterium grows to logarithmic phase, and is standby;
(3) fermentation culture of bacterial classification: 100 μ l insert in step (1) the DF substratum with step (2) activation bacteria suspension, cultivate 24-48h down at 25 ℃-37 ℃, to thalline logarithmic growth after date, carry out centrifugation, obtain thalline, and are standby;
(4) draw seed pelleting of eggplant: get step (3) bacteria suspension 0.5ml, with the 0.03M MgSO of sterilization
4Resuspension, and dilution 8-10 is doubly, measures to absorption value 〉=0.1 at the 600nm place; That gets that embodiment 3 filters out draws seed of eggplant with 70% Ethanol Treatment 1min, and sterilized water is cleaned; Handle 10min with 1%NaClO again, sterilized water is cleaned; With seed immersion treatment 1h-2h in bacteria suspension, make the seed-coat bacterial content reach 3.0 * 10
7-6.0 * 10
7CFU/;
(5) growing vegetables: add salinity and fertilizer on request, mixing dress basin.Every basin is adorned native 1kg, and adding distil water makes soil moisture content reach 75% of field capacity, leaves standstill.Every basin adds urea 0.1722g, potassium primary phosphate 0.0376g, vitriolate of tartar 0.1164g, accurately adds required salinity, makes soil salt content reach 1.0g/kg, 2.0g/kg, 3.0g/kg NaCl respectively.Each is handled 3 times and repeats.Seed is sowed after treatment.Final singling after 1 month, the upgrowth situation of mensuration plant after 2 months, fresh weight, ion content and the proline content of mensuration eggplant seedling.
The result shows, handles by having 1-amino-cyclopropane 1-carboxylic acid deaminase active bacterial strain, can significantly increase the fresh weight of eggplant seedling under 0.1% and 0.2% salt concn, increases Ca
2+And k
+Ionic content reduces proline content and Na
+Ion content (seeing Table 2).
Table 2 draws the measurement result after eggplant connects the bacterium processing for No.
Proline content
Bacterium is handled salt concn/% fresh weight/g Na/% K/% Ca/ ‰/‰
- 0 16.16 0.775 3.112 3.318 0.332
+ 0 17.81 0.74 3.215 4.01 0.197
- 0.1 20.99 0.932 3.096 2.767 0.359
+ 0.1 23.63 0.498 3.112 2.96 0.298
- 0.2 10.8 1.68 2.248 2.272 1.415
+ 0.2 13.39 1.283 2.728 2.44 1.018
- 0.3 5.23 3.06 1.724 1.921 1.662
+ 0.3 7.3 2.494 2.023 2.427 1.327
Annotate :-(blank)
The structure and the potted plant experiment of the assorted tomato-microflora in embodiment 5:(Zhejiang)
(1) DF culture medium preparation; (2) activation culture of bacterial classification; (3) fermentation culture of bacterial classification; (4) the assorted tomato seeds dressing in Zhejiang; And (5) growing vegetables: method is measured the upgrowth situation of plant with embodiment after 4,2 months, fresh weight and the antioxidase system activity of mensuration seedling.
The result shows, handles by having 1-amino-cyclopropane 1-carboxylic acid deaminase active bacterial strain, can significantly increase the fresh weight of tomato seedling under 0.1% and 0.2% salt concn, increases activities of antioxidant enzymes in the plant body.
The assorted tomato in table 3 Zhejiang connects the measurement result after bacterium is handled
Salt concn fresh weight POD/ (u CAT/ (u APX/ (u
Bacterium processing/%/g protein
-1Min
-1) protein
-1Min
-1) protein
-1Min
-1)
- 0 35.37?3.551 4.829 0.612
+ 0 35.35?3.952 8.861 0.784
- 0.1 25.21?1.525 5.766 0.421
+ 0.1 28.59?2.5 10.07 0.685
- 0.2 23.92?1.361 6.662 0.352
+ 0.2 26.21?1.829 11.789 0.425
- 0.3 20.52?0.849 6.142 0.113
+ 0.3 22.81?1.041 8.746 0.359
Sequence table
＜110〉Zhejiang Academy of Agricultural Science
＜120〉a kind of method that strengthens the vegetable crop salt tolerance
<160>2
<210>1
<211>20
<212>DNA
＜213〉primer
<400>1
AGAGTTTGAT?CCTGGCTCAG?20
<210>2
<211>20
<212>DNA
＜213〉primer
<400>2
AAGGAGGTGA?TCCAGCCGCA?20
Claims (2)
1, pseudomonas (Pseudomonas sp.) bacterial strain DW1, its bacterial strain deposit number is CGMCCNo.2729.
2, a kind of method that strengthens the vegetable crop salt tolerance is characterized in that this method carries out according to the following steps:
(1) culture medium preparation: the DF substratum, every L contains KH
2PO
44.0g, Na
2HPO
46.0g, MgSO
47H
2O 0.2g, glucose 2.0g, gluconic acid 2.0g, citric acid 2.0g, (NH
4)
2SO
42.0g, pH 7.2-7.4; The DFa substratum joins ACC and does not contain (NH
4)
2SO
4The DF substratum in, its final concentration is 〉=2.0mmolL
-1, after the sterilization, standby;
(2) activation culture of bacterial classification: with pseudomonas (Pseudomonas sp.) bacterial strain DW1, CGMCCNo.2729 is inoculated in the DFa substratum and activates, and treats that bacterium grows to logarithmic phase, and is standby;
(3) fermentation culture of bacterial classification: 100 μ l insert in step (1) the DF substratum with step (2) activation bacteria suspension, cultivate 24-48h down at 25 ℃-37 ℃, to thalline logarithmic growth after date, carry out centrifugation, obtain thalline, and are standby;
(4) screening of salt tolerant vegetable variety:
A, to the comparison of various vegetable crop seed germination rates, filter out the vegetable variety that higher germination rate is arranged under condition of salt stress;
B, the vegetable variety higher to the seed germination rate that filters out among the step a carry out the experiment of growth of seedling salt tolerance, measure by plant growth condition, therefrom filter out the vegetable variety of salt tolerant;
(5) structure of vegetable variety-microflora: it is A that step (3) fermentation thalline is mixed with concentration
600〉=0.1 bacteria suspension; After the salt tolerant vegetable variety seed disinfection processing that step (4) is filtered out, sterilized water are cleaned, in bacteria suspension, soak, dressing, make the seed-coat bacteria containing amount reach 3.0 * 10
7-6.0 * 10
7CFU/, can sow after draining, plant.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101625527A CN101492647A (en) | 2008-12-01 | 2008-12-01 | Method for reinforcing salt endurance of vegetable crop |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CNA2008101625527A CN101492647A (en) | 2008-12-01 | 2008-12-01 | Method for reinforcing salt endurance of vegetable crop |
Publications (1)
| Publication Number | Publication Date |
|---|---|
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Family
ID=40923439
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|---|---|---|---|
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Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103607884A (en) * | 2011-03-17 | 2014-02-26 | 生物探索（新西兰）有限公司 | Screening methods |
| CN103875469A (en) * | 2012-12-20 | 2014-06-25 | 盐城工学院 | Method for quickly evaluating salt tolerance of grapes |
| CN108440185A (en) * | 2018-04-20 | 2018-08-24 | 浙江省农业科学院 | A kind of biological nutrition type water-retaining agent and preparation method thereof |
-
2008
- 2008-12-01 CN CNA2008101625527A patent/CN101492647A/en active Pending
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103607884A (en) * | 2011-03-17 | 2014-02-26 | 生物探索（新西兰）有限公司 | Screening methods |
| CN103875469A (en) * | 2012-12-20 | 2014-06-25 | 盐城工学院 | Method for quickly evaluating salt tolerance of grapes |
| CN108440185A (en) * | 2018-04-20 | 2018-08-24 | 浙江省农业科学院 | A kind of biological nutrition type water-retaining agent and preparation method thereof |
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