CN103875398B - A kind of method that reduces chromium content in tobacco leaf - Google Patents
A kind of method that reduces chromium content in tobacco leaf Download PDFInfo
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- CN103875398B CN103875398B CN201410092267.8A CN201410092267A CN103875398B CN 103875398 B CN103875398 B CN 103875398B CN 201410092267 A CN201410092267 A CN 201410092267A CN 103875398 B CN103875398 B CN 103875398B
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Abstract
Reduce a method for tobacco leaf chromium content, it is residual that utilization reduces tobacco heavy metal chromium at the parasitic India of tobacco root pyriform spore. The concrete steps of described method are as follows: (1) replaces the ZnSO in former Kafer culture medium with nano material zinc oxide4·7H2O adds nano material zinc oxide 10mg in 1 liter of culture medium; (2) by the India pyriform spore (Piriformospora after activation? indica) be inoculated in the described liquid Kafer culture medium of step (1) and cultivate, then collect mycelium, preparation India pyriform spore mycelia liquid, then water in cigarette shoot root portion, root irrigation, grows surely in cigarette shoot root portion India's pyriform spore; (3) after India's pyriform spore is grown surely, cigarette transplantation of seedlings, to land for growing field crops, is carried out conventional water and fertilizer management afterwards, makes tobacco normal growth. This method can be improved tobacco growing situation, reduces the migration of heavy metal chromium from root system to aboveground part of tobacco at its, reduces tobacco leaf heavy metal chromium residual, and be easy to grasp, cost is lower, simple and practical, environmental protection, can not cause secondary pollution.
Description
Technical field
The present invention relates to utilization and the agricultural product quality and safety technical field of microbial resources, outstandingIt relates to a kind of method that reduces tobacco leaf chromium content.
Background technology
Along with using in a large number of chemical fertilizer, agricultural chemicals and developing rapidly of industry, environmental pollution is day by day tightHeavy. Chromium is a kind of poisonous heavy metal, is also one of main environmental contaminants. Tobacco is for IOne of industrial crops that state is important, cultivated area, yield of flue-cured tobacco and the output of China tobacco are equalOccupy first place in the world. In recent years, often there is the report of the quality security problems such as tobacco heavy metals exceeding standard,Wherein cadmium, chromium, plumbous comparatively serious on the impact of tobacco growing and quality. Pollution of chromium in soilThe hysteresis that causes the slow of tobacco growing and grow, has reduced total nitrogen in tobacco, nicotine, solvableProperty total reducing sugar and reduced sugar content, had a strong impact on tobacco leaf quality (Shi Hongzhi, Liu Guoshun, normalThink quick, etc. tobacco heavy metal present Research and agriculture harm reduction countermeasure. Chinese tobacco journal,2011,17 (3): 89-94). Heavy metal becomes the key factor that affects China's Flue-cured tobacco Quality securityOne of, the 9th the Asia-Pacific tobacco and strong of holding at Sydney, AUS on October 7th, 2010On Kang Huiyi, issue the result of study of a China and other countries' tobacco contrast: China 13The cigarette of individual plate detects the content of beary metal such as chromium and Canadian cigarette comparison, the highest exceedingMore than 3 times. In tobacco leaf, chromium content overproof has had a strong impact on the quality of tobacco leaf, certainly will have influence on meThe competitiveness of state's tobacco industry and sound development. Therefore, how reducing chromium in tobacco leaf has accumulatedBecome main task of tobacco business.
In tobacco, heavy metal is mainly from tobacco-growing soil, and fertilizer and the lack of standardization of agricultural chemicals also can simultaneouslyCause the content of beary metal of tobacco too high. The major measure of administering tobacco-growing soil heavy metal pollution hasThe methods such as engineering measure, physical chemistry reparation, phytoremediation and rhizosphere microorganism reparation, but onState operability, validity, broad spectrum activity and security that method is repaired heavy metal pollution of soilAll be left to be desired and improve. At present, the measure of the most frequently used reduction tobacco heavy-metal residual is to useVarious modifying agents, as inorganic amendments such as lime, calcium magnesium phosphate, clay minerals, and livestock and poultryThe various fertilizers such as ight soil etc. (Hu Zhongsheng, Zhang Gangya, Wang Guangzhi, etc. renovation agent absorbs dirty to tobaccoDye the improvement of Arsenic in Soil. soil, 2006,38 (2): 200-205; Zhou Zeqing, recruits and opens cypress, Zhu WeiStar, etc. heavy metal pollution tobacco-planting soil conditioner original position is repaired progress. Agriculture of Anhui sectionLearn 2009,37 (11): 5100-5102.). Chinese invention patent (CN1017M405A) has been inventedReduce a soil conditioner for crops cadmium, lead contamination, mainly comprise Ca (OH)2、Na2SiO7Deng organic components such as inorganic component and rice straw, mountain green soy bean casts. For various reasons,The large area of these modifying agents is often used and is restricted, if lime, clay mineral are to soilThe impact of shape is larger, and often contains heavy metal in calcium magnesium phosphate, feces of livestock and poultry. Therefore, carryThe technical measures that go out a kind of more economical practicality just seem for effectively reducing tobacco heavy-metal residualVery necessary.
Chinese patent application 201310104049.7 discloses trichoderma reesei and has combined companion's ore deposit red-spotted stonecrop and existApplication in the reparation of cadmium pollution agricultural land soil, microbial inoculum using method is at companion ore deposit red-spotted stonecrop seedling rhizosphereApply trichoderma reesei FS10-C zymotic fluid or microorganism formulation; Chinese patent application201210137704 disclose a kind of method of utilizing arbuscular mycorrhiza restoration of soil polluted by heavy metal,Bacterial classification using method is that arbuscular mycorrhiza is inoculated in to barnyard grass root. Above-mentioned two patents are with biologicalBacterial classification improves host and absorbs the ability of heavy metal in soil, removes subsequently host plant, completesThe reparation of heavy-metal contaminated soil. These class methods can reduce the content of beary metal in soil, butAlso reduced the multiple crop index of ploughing, and how to process host plant be also in production reality simultaneouslyA difficult problem. Chinese patent application 20121007120 and 201010525189 all discloses oneUtilize AMF to reduce method, AMF and the tobacco of tobacco heavy-metal residualForm arbuscular mycorrhiza, promote tobacco growing, reduce tobacco heavy-metal residual. But clumpMycorrhizal fungi can not be grown on artificial synthetic culture medium, is unfavorable for pushing away on a large scale of this bacteriumWide application.
India's pyriform spore is a kind of root endogenetic fungus, has host range widely, can promotePlant growth, improves the resistance of host to biological and abiotic stress, and can be at synthetic mediaUpper cultivation, for this bacterium agricultural and the large-area applications of horticulture provide possible (Lou Binggan,Sun Chao, Cai great Guang. several functions and the application prospect thereof of India's pyriform spore. plant protection journal.2007,34 (6): 653-656). Therefore,, by developing India's pyriform spore, do not affecting cigaretteUnder the prerequisite of leaf-making quantity and quality, reduce the content of content of beary metal in tobacco leaf, especially chromium, toolHave wide practical use.
Summary of the invention
The problem existing for solving above-mentioned prior art, the object of the present invention is to provide one to fallThe method of cadmium content in low tobacco leaf, utilizes biological method to reduce in tobacco leaf heavy metal chromium residual.
To achieve these goals, technical scheme of the present invention is:
Reduce a method for chromium content in tobacco leaf, utilize the pyriform spore in the parasitic India of tobacco rootReduce tobacco heavy metal chromium residual.
Further, the method for described reduction tobacco leaf chromium content specifically comprises the following steps:
(1) preparation of India's pyriform spore culture medium:
The nano material zinc oxide that is 30nm with particle diameter replaces in former Kafer culture mediumZnSO4·7H2O adds nano material zinc oxide 10mg in 1 liter of culture medium, describedly newly joinsIn the Kafer culture medium of putting, every liter contains: 2.00g peptone, and 1.00g yeast extract,1g caseinhydrolysate, 0.6gNaNO3，0.52gKCl，0.52gMgSO4·7H2O，1.52gKH2PO4，10mgZnO，10mgH3BO3，5mgMnSO4·H2O，1.6mgCoCl·5H2O，1.6mgCuSO4·5H2O，1.1mg(NH4)6MO7O24，1.3mgFeSO4，1.8mgEDTA, 0.1mg thiamine, 0.04mg glycine, 0.01mg nicotinic acid and 0.01mgPyridoxol;
(2) India's pyriform spore mycelia liquid irrigating root is processed cigarette shoot root portion
India's pyriform spore after activation is inoculated in to the described liquid Kafer of step (1) to be cultivatedCellar culture in base, then collects mycelium, and preparation India pyriform spore mycelia liquid, waters in cigaretteShoot root portion, carries out root irrigation, and India's pyriform spore is grown surely in cigarette shoot root portion;
(3) transplanting of tobacco seedling and the management in later stage
Tobacco seedling after India's pyriform spore is grown is surely transplanted, and carries out afterwards conventional liquid manure pipeReason, makes cigarette seedling normal growth.
Further, in described step (2), the condition of culture of India's pyriform spore is 25 DEG C-28DEG C, 100rpm-120rpm shaking table lucifuge cultivate, after 5-7d, filter, collect mycelium, with steamHeat up in a steamer water and be made into the mycelia liquid that concentration is 2g/L-4g/L.
Further, in step (2), the mode of India's pyriform spore mycelia liquid processing tobacco seedling is:India's pyriform spore mycelia liquid is watered in tobacco seedling root; Or India's pyriform spore mycelia liquid is addedTobacco seedling device is grown seedlings to tobacco; Or tobacco seed is carried out with India's pyriform spore mycelia liquidSeed pelleting, can also be stained with root processing to tobacco seedling with India's pyriform spore mycelia liquid.
Further, in step (2), water in tobacco seedling root with India's pyriform spore mycelia liquidTime every seedling pouring India's pyriform spore mycelia liquid measure be 50mL.
Further, in step (2), cigarette seedling is that 4 Ye Qishiyong India pyriform spore mycelia liquid waterFill with in tobacco seedling root.
With respect to prior art, beneficial effect of the present invention is:
1,, compared with AMF in prior art, India of the present invention pyriform spore can beOn culture medium, grow, application is convenient; And the nutrition condition that can improve tobacco, has bio-fertilizerThe function of material, promotes the growth of tobacco, reduces the use of chemical fertilizer.
2, to adopt particle diameter be that the nano material zinc oxide of 30nm replaces former Kafer to cultivate in the present inventionZnSO in base4·7H2O adds nano material zinc oxide 10mg in 1 liter of culture medium,Contain India's pyriform spore of the Kafer medium culture of nano material ZnO (30nm) 0.01mg/L,Its rate of growing of determining at tobacco root significantly improves, and brings up to 95.0%, energy by original 80.0%Improve significantly biological tobacco amount, promote the growth of tobacco, reduce heavy metal from root system to tobaccoThe migration of aerial part, thus it is residual to reduce tobacco heavy metal chromium. Test confirms to use the present inventionMethod can reduce chromium content in Mature Tobacco Leaves significantly, makes water planting, potted plant and land for growing field crops Mature Tobacco LeavesThe content of middle chromium decline respectively 31.4-54.2%, 27.9-41.3%, 32.9-55.3%.
3, the present invention reduce in tobacco that the method for heavy-metal residual is easy to grasp, cost is lower,Simple and practical, environmental protection, can not cause secondary pollution; It is not only applicable to heavy metal pollutionThe plantation of tobacco in soil, also can be used for uncontamination soil.
4, in the present invention, the using method of India's pyriform spore mycelia liquid is various, can be by India's pearsShape spore mycelia liquid waters in tobacco seedling root, can also add tobacco seedling device to educate tobaccoSeedling, can also carry out seed pelleting to tobacco seed with India's pyriform spore mycelia liquid, can also be rightTobacco seedling is stained with root processing.
Detailed description of the invention
Below in conjunction with specific embodiment, the present invention program is described in further details:
Reduce a method for chromium content in tobacco leaf, utilize the pyriform spore in the parasitic India of tobacco rootReduce tobacco heavy metal chromium residual, India of the present invention pyriform spore is by the Ajit of Ai De university of IndiaProfessor Varma is so kind as to give.
Further, the method for described reduction tobacco leaf chromium content specifically comprises the following steps:
(1) preparation of India's pyriform spore culture medium:
The nano material zinc oxide that is 30nm with particle diameter replaces in former Kafer culture mediumZnSO4·7H2O adds nano material zinc oxide 10mg in 1 liter of culture medium, describedly newly joinsIn the Kafer culture medium of putting, every liter contains: 2.00g peptone, and 1.00g yeast extract,1g caseinhydrolysate, 0.6gNaNO3，0.52gKCl，0.52gMgSO4·7H2O，1.52gKH2PO4，10mgZnO，10mgH3BO3，5mgMnSO4·H2O，1.6mgCoCl·5H2O，1.6mgCuSO4·5H2O，1.1mg(NH4)6MO7O24，1.3mgFeSO4，1.8mgEDTA, 0.1mg thiamine, 0.04mg glycine, 0.01mg nicotinic acid and 0.01mgPyridoxol;
(2) India's pyriform spore mycelia liquid irrigating root is processed cigarette shoot root portion
India's pyriform spore after activation is inoculated in to the described liquid Kafer of step (1) to be cultivatedCellar culture in base, then collects mycelium, and preparation India pyriform spore mycelia liquid, waters in cigaretteShoot root portion, carries out root irrigation, and India's pyriform spore is grown surely in cigarette shoot root portion;
(3) transplanting of tobacco seedling and the management in later stage
Tobacco seedling after India's pyriform spore is grown is surely transplanted, and carries out afterwards conventional liquid manure pipeReason, makes cigarette seedling normal growth.
Further, in described step (2), the condition of culture of India's pyriform spore is 25 DEG C-28DEG C, 100rpm-120rpm shaking table lucifuge cultivate, after 5-7d, filter, collect mycelium, with steamHeat up in a steamer water and be made into the mycelia liquid that concentration is 2g/L-4g/L.
Further, in step (2), the mode of India's pyriform spore mycelia liquid processing tobacco seedling is:India's pyriform spore mycelia liquid is watered in tobacco seedling root; Or India's pyriform spore mycelia liquid is addedTobacco seedling device is grown seedlings to tobacco; Or tobacco seed is carried out with India's pyriform spore mycelia liquidSeed pelleting, can also be stained with root processing to tobacco seedling with India's pyriform spore mycelia liquid.
Further, in step (2), water in tobacco seedling root with India's pyriform spore mycelia liquidTime every seedling pouring India's pyriform spore mycelia liquid measure be 50mL.
Further, in step (2), cigarette seedling is that 4 Ye Qishiyong India pyriform spore mycelia liquid waterFill with in tobacco seedling root.
Embodiment 1
Utilize India's pyriform spore to reduce water planting tobacco heavy metal chromium residual, comprise the steps:
The activation of bacterial classification: the India's pyriform spore bacterial classification being kept in refrigerator is inoculated in to Kafer trainingSupport on base (KM) flat board, be placed in lucifuge in 28 DEG C of insulating boxs and cultivate 7 days. With diameter be 5mmThe India pyriform spore colony edge punching that activating of card punch.
The cultivation of bacterium liquid: obtaining liq Kafer culture medium and containing nano material ZnO (30nm)Liquid Kafer culture medium (the i.e. nano material ZnO generation taking particle diameter as 30nm of 0.01g/LFor ZnSO4), with the packing of 250mL triangular pyramidal bottle, every bottled 150mL culture medium, goes outBacterium is for subsequent use. Under aseptic condition, India pyriform spore mycelia piece is added above-mentioned culture medium is housedIn triangular pyramidal bottle, every bottle of 6 ferfas silk pieces; Be placed in constant-temperature table 120rpm, 28 DEG C of lucifugesShaken cultivation 5-7 days.
Mycelial collection: in conical flask, mycelia is placed in 250mL centrifugal bottle, 8000rpmCentrifugal 20min, removes supernatant, collects mycelium, is placed in 4 DEG C of preservations, for subsequent use;
The preparation of cigarette seedling: by sowing in the seedlings nursing plate of hole after tobacco seed sterilization, treat that seed goes outThinning after seedling, 1 of every Kong Liumiao, carries out normal water and fertilizer management.
Mycelia liquid is poured onto rice shoot root: water is made into India pyriform spore mycelia containing mycelial concentrationFor the mycelia liquid of 4g/L, by mycelia liquid water in 4 the leaf phase cigarette shoot root portion, establish 2 processing and 1Individual contrast. Processing 1 is the liquid Kafer medium culture containing nano material ZnO (30nm)India's pyriform spore mycelia liquid, processing 2 is India's pyriform of common liq Kafer medium cultureSpore mycelia liquid, the water of contrast pouring same volume.
Mycelia liquid waters latter 15 days, detects India's pears by the micro-Microscopic observation of Trypan BlueShape spore is in the situation of growing of determining of cigarette shoot root portion, and the rate of growing of determining of result Graphics Processing 1 is 93%, processes2 the rate of growing of determining is 81%, and result shows the liquid Kafer training containing nano material ZnO (30nm)Supporting the India pyriform spore mycelia of base cultivation obviously improves in the rate of growing of determining of cigarette shoot root portion.
(1) cigarette seedling is moved to and in Plastic Drum, carries out water planting: select process 1 and contrast in growing wayConsistent cigarette transplantation of seedlings, to Plastic Drum (volume is 4L), is equipped with chromium concn for containing in bucketThe Hoagland nutrient solution 4L of 5.0mg/L, cultivates 4 strains of cigarette seedling for every barrel, and contrast and processing are each3 barrels.
Measure the chromium content in tobacco leaf: sampling (maximum leaf) in 2 weeks after water planting, measure chromium elementContent, result is that processed group tobacco leaf chromium content has reduced 43.1%(table 1 than contrast).
Table 1 chromium is processed chromium content in the water planting tobacco leaf of inoculating and do not inoculate India's pyriform spore for 2 weeks
Note: process the 1 India's pyriform spore mycelia liquid for the liquid Kafer medium culture containing nano material ZnO (30nm) (table2, table 3 is same)
(2) cigarette seedling is moved in flowerpot, carry out potted plant: select process 1 and contrast in growing way oneThe cigarette transplantation of seedlings causing is (end diameter 20cm, high 27cm) to plastic tub, in basin, is equipped with and containsCr(NO3)2Concentration is the field soil 10kg of 200mg/kg, and 1 strain of every basin plantation cigarette seedling is rightAccording to process each 3 strains.
Measure the chromium content in tobacco leaf: sampling (maximum leaf) in potted plant latter 50 days, measure chromium elementContent, result is that processed group tobacco leaf chromium content has reduced 27.9%(table 2 than contrast).
Table 2 chromium is processed the content of rear Tobacco Root and Ye Zhongge
(3) cigarette seedling is moved to large Tanaka's cultivation: select processing 1 consistent with growing way in contrastCigarette transplantation of seedlings, to large Tanaka, is carried out chromium Stress treatment after 70 days: centered by tobacco stem base portion,10cm is radius, and the chromium nitrate solution 500mL that pouring concentration is 4.0g/L, to water 500mLWater in contrast. Each 10 strains of each processing; The climax leaves of gathering after 50 days, measure in tobacco leafThe content of chromium.
Measure the chromium content in tobacco leaf: chromium is processed the Mature Tobacco Leaves of gathering after 50 days, measures chromium unitCellulose content, result is that processed group tobacco leaf chromium content has reduced 32.9%(table 3 than contrast).
Table 3 chromium is processed chromium content in the Mature Tobacco Leaves of rear land for growing field crops
Embodiment 2
Make India's pyriform spore mycelia piece in embodiment 1 into India pyriform spore spore liquid, be inoculated inContaining in the liquid Kafer culture medium of 10mg/LZnO, keep away in constant-temperature table 120rpm, 25 DEG CLight generation is cultivated 7 days, and all the other are equal to example 1, final water planting, potted plant and field production cigaretteThe content of Ye Zhongge declines respectively 54.2%, 41.2%, 51.4%.
Embodiment 3
Change India's pyriform spore mycelia liquid concentration in embodiment 1 into 2g/L, all the other are equal to realityExample 1, in final water planting, potted plant and field production tobacco leaf the content of chromium decline respectively 50.2%,41.0%、50.9%。
Embodiment 4
India's pyriform spore mycelia liquid in embodiment 1 is watered in tobacco seedling root and makes India's pears intoShape spore mycelia liquid adds tobacco seedling device to grow seedlings to tobacco, and all the other are equal to example 1,In whole water planting, potted plant and field production tobacco leaf the content of chromium decline respectively 31.4%, 40.9%,31.3%。
Embodiment 5
India's pyriform spore mycelia liquid in embodiment 1 is watered in tobacco seedling root and makes India's pears intoShape spore mycelia liquid is stained with root processing to tobacco seedling, and all the other are equal to example 1, final water planting,In potted plant and field production tobacco leaf, the content of chromium declines respectively 52.2%, 41.3%, 55.3%.
The above be only the specific embodiment of the present invention, but protection scope of the present invention alsoBe not limited to this, any variation of expecting without creative work or replacement, all should be encompassed inWithin protection scope of the present invention. Therefore, protection scope of the present invention should be with claimsThe protection domain limiting is as the criterion.
Claims (5)
1. a method that reduces chromium content in tobacco leaf, is characterized in that, utilizes in the parasitic India of tobacco rootIt is residual that pyriform spore reduces tobacco heavy metal chromium,
The method of described reduction tobacco leaf chromium content specifically comprises the following steps:
(1) preparation of India's pyriform spore culture medium:
The nano material zinc oxide that is 30nm with particle diameter replaces the ZnSO in former Kafer culture medium4·7H2O，In 1 liter of culture medium, add nano material zinc oxide 10mg, every in the Kafer culture medium of described new configurationRise and contain: 2.00g peptone, 1.00g yeast extract, 1g caseinhydrolysate, 0.6gNaNO3，0.52gKCl，0.52gMgSO4·7H2O，1.52gKH2PO4，10mgZnO，10mgH3BO3，5mgMnSO4·H2O，1.6mgCoCl·5H2O，1.6mgCuSO4·5H2O，1.1mg(NH4)6MO7O24，1.3mgFeSO4, 1.8mgEDTA, 0.1mg thiamine, 0.04mg glycine,0.01mg nicotinic acid and 0.01mg pyridoxol;
(2) India's pyriform spore mycelia liquid irrigating root is processed cigarette shoot root portion
India's pyriform spore after activation is inoculated in the Kafer culture medium of the described new configuration of step (1)Cellar culture, then collects mycelium, and preparation India pyriform spore mycelia liquid, waters in cigarette shoot root portion, entersRow root irrigation, grows surely in cigarette shoot root portion India's pyriform spore;
(3) transplanting of tobacco seedling and the management in later stage
Tobacco seedling after India's pyriform spore is grown is surely transplanted, and carries out afterwards conventional water and fertilizer management, makes cigaretteSeedling normal growth.
2. the method for claim 1, is characterized in that, India's pyriform spore in described step (2)Condition of culture be that 25 DEG C-28 DEG C, 100rpm-120rpm shaking table lucifuge are cultivated, after 5-7d, filter, collectMycelium, is made into the mycelia liquid that concentration is 2g/L-4g/L with distilled water.
3. the method for claim 1, is characterized in that, India pyriform spore bacterium in step (2)The mode that silk liquid is processed tobacco seedling is: India's pyriform spore mycelia liquid is watered in tobacco seedling root; Maybe will printDegree pyriform spore mycelia liquid adds tobacco seedling device to grow seedlings to tobacco; Or with India's pyriform spore mycelia liquid pairTobacco seed carries out seed pelleting, or tobacco seedling is stained with to root processing with India's pyriform spore mycelia liquid.
4. method as claimed in claim 3, is characterized in that, uses India's pyriform spore in step (2)India's pyriform spore mycelia liquid measure that mycelia liquid waters every seedling pouring in the time of tobacco seedling root is 50mL.
5. method as claimed in claim 3, is characterized in that, when in step (2), cigarette seedling was 4 leaf phaseWater in tobacco seedling root with India's pyriform spore mycelia liquid.
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| CN111849856A (en) * | 2020-07-15 | 2020-10-30 | 江苏里下河地区农业科学研究所 | Indoca chlamydospore, P.indoca spore bacterial agent and preparation method thereof |
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| CN102577843A (en) * | 2012-03-13 | 2012-07-18 | 中国烟草总公司郑州烟草研究院 | Method for lowering Cd content in tobacco leaves by utilizing grafting technology |
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