CN102948399A - Daphnia magna breeding method for evaluating safety of transgenic rice - Google Patents
Daphnia magna breeding method for evaluating safety of transgenic rice Download PDFInfo
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- CN102948399A CN102948399A CN2012101069819A CN201210106981A CN102948399A CN 102948399 A CN102948399 A CN 102948399A CN 2012101069819 A CN2012101069819 A CN 2012101069819A CN 201210106981 A CN201210106981 A CN 201210106981A CN 102948399 A CN102948399 A CN 102948399A
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Abstract
The invention relates to a breeding method for an aquatic organism daphnia magna, and belongs to the technical field of insect breeding. The method is used for overcoming the defect in a traditional daphnia magna toxicity experiment technology, and the invention builds a method for breeding daphnia magna by utilizing rice. The method comprises the following steps of: preparing a nutrient solution, preparing a rice flour solution, breeding and collecting young fleas. According to the daphnia magna breeding method disclosed by the invention, rice is used for breeding the daphnia magna, so that the defect that the daphnia magna in the conventional method selectively takes in natural goods such as green alga, but does not take in or take in a little rice is overcome. Moreover, an adverse factor that the experimental result is easily affected by uncertain nutritional ingredients of the green alga, and reliable experiment support is provided for evaluating the safety of transgenic rice.
Description
Technical field:
The present invention relates to a kind of raising method of aquatile Daphnia magna, belong to technical field of insect feeding
Background technology:
Paddy rice is as one of important cereal crops in the world, and for nearly 3,000,000,000 people on the earth are providing food, but damage by disease and insect has a strong impact on output and the quality of paddy rice.By transgenic technology the foreign gene such as pest-resistant, disease-resistant is incorporated in the rice genome, cultivates pest-resistant, disease-resistant transgenic new rice variety, become new method and new way that the prevention and control rice pest improves rice yield.Along with the large-scale promotion of genetically modified crops, people more and more pay close attention to its safety issue.Because China's Rice Production and the critical role and the impact that are consumed in the whole world; very careful and conscientious attitude is held for commercially producing of transgenic paddy rice by the Chinese government; both considered the following commercialization of transgenic paddy rice with the tremendous economic interests and the social influence that bring, the while is also carried out science and conscientious evaluation to the potential source biomolecule safety problem that the extensive environment of transgenic paddy rice discharges and plantation may bring.
Daphnia magna (Daphnia magna) is the standard biological in the ecotoxicology research, is widely used in the safety evaluation of various chemicals toxicity.Daphnia magna is distributed widely in the various water bodys, comprises the paddy field of rice cultivation, and the environmental safety assessment of therefore carrying out transgenic paddy rice take Daphnia magna as research object is of practical significance very much.Mainly be artificial cultivation and the toxicological evaluation that carries out Daphnia magna take green alga (such as scenedesmus obliquus Scenedesmus obliquus) as Major Foods at present, can't estimate exactly this class insoluble solid of transgenic rice to the true impact of Daphnia magna.In order to estimate more accurately transgenic paddy rice to the toxicological effect of Daphnia magna, in the urgent need to a kind of Daphnia magna raising method take rice paddy seed as the Major Foods composition.
Summary of the invention:
The present invention has overcome the defective of traditional Daphnia magna toxicological experiment technology, has set up the method for the pure rice raising of cover Daphnia magna, provides reliable experiment guarantee for estimating the transgenic paddy rice safety, and method step is as follows:
Ground rice solution preparation: after rice paddy seed shelled, fully smash rear mistake 125 mesh sieves with cracker, make the ground rice particle diameter less than 100um, take by weighing the powder after sieving, be configured to concentration with culture fluid and be respectively 0.8gL
-1, 1.2gL
-1, 1.6gL
-1Ground rice suspension, minute be filled to behind the homogeneous in the 10ml sterilization centrifuge tube, 4 ℃ store for future use.
Beneficial effect of the present invention
(1) the present invention uses pure rice to raise Daphnia magna, has eliminated the natural food such as the medium-and-large-sized flea selectivity picked-up of conventional method green alga and does not take food or seldom take food the phenomenon of rice.
(2) the present invention uses pure rice to feed Daphnia magna, and having overcome in the conventional method easily affects experimental result because the green algae nutrient composition is uncertain, and needs constantly to cultivate the defective of green alga.
Description of drawings:
Fig. 1: transgenic paddy rice and the parent survival fate under variable concentrations rice is fed relatively
The impact that Fig. 2 different disposal is long on the Daphnia magna body
Fig. 3 different disposal is on the impact of Daphnia magna survival rate
Embodiment:
Feed the biological safety of raising Daphnia magna evaluation transgenic paddy rice with pure rice.It is " Bt Shanyou 63 " and parental seed thereof that rice paddy seed is selected in test, and the foreign gene of insertion is Cry1Ab/Cry1Ac pattern of fusion gene.
Daphnia magna (Daphnia magna) is after the long-term cultivation domestication, and the young flea of 6～24h that selection above genetic background of 3 generations of cultivation is identical is as the experiment flea.The experiment flea is 22 ± 1 ℃ in temperature, and light intensity is 60 μ mol m-2s-1, and Light To Dark Ratio is to cultivate in the constant temperature culture chamber of 16h: 8h.Experiment culture fluid prescription: 294.0mgL-1CaCl2; 123.2mgL-1MgSO4; 64.8mgL-1NaHCO3; 6.3mgL-1KCL the culture fluid pH of new preparation is 7.8 ± 0.2, hardness 250 ± 25mgL-1 (in CaCO3).Each processed group is added the ground rice solution 100ul of variable concentrations every day, (0.8gL-1,1.2gL-1,1.6gL-1 ground rice, dry weight is equivalent to respectively 2,3,4mg, this concentration is required every day with enough water fleas) in the incubation every day regularly clear up foreign material and the little water flea of new life shifted out, changed in per 3 days and to cultivate water.
In the feeding process, measure respectively body length, survival rate and the survival fate of different disposal group Daphnia magna:
Can find out that from the above results transgenic paddy rice has no significant effect with the growth that its parent compares Daphnia magna, because this raising method nutrient component is all clear and definite controlled, got rid of the impact of uncertain factor on experiment, so the result be more rigorous credible.
Claims (4)
1. a Daphnia magna raising method of estimating the transgenic paddy rice safety is characterized in that culture fluid preparation: 294.0mgL
-1CaCl2; 123.2mgL
-1MgSO4; 64.8mgL
-1NaHCO3; 6.3mgL
-1KCL, the culture fluid pH of new preparation is 7.8 ± 0.2, hardness 250 ± 25mgL
-1(in CaCO3).
2. Daphnia magna raising method of estimating the transgenic paddy rice safety, it is characterized in that the ground rice solution preparation: after rice paddy seed is shelled, fully smash rear mistake 125 mesh sieves with cracker, make the ground rice particle diameter less than 100um, take by weighing the powder after sieving, be configured to concentration with culture fluid and be respectively 0.8gL
-1, 1.2gL
-1, 1.6gL
-1Ground rice suspension, minute be filled to behind the homogeneous in the 10ml sterilization centrifuge tube, 4 ℃ store for future use.
3. Daphnia magna raising method of estimating the transgenic paddy rice safety, it is characterized in that testing flea is 22 ± 1 ℃ in temperature, and light intensity is 60 μ mol m-2s-1, and Light To Dark Ratio is to cultivate in the constant temperature culture chamber 50ml beaker of 16h: 8h, and every glass adds the 40ml culture fluid.
4. a Daphnia magna raising method of estimating the transgenic paddy rice safety is characterized in that adding ground rice solution 100ul every day, and regularly clear up foreign material and the little water flea of new life is shifted out per 3 days replacing culture fluids every day in the incubation.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782576A (en) * | 2015-04-13 | 2015-07-22 | 临沂大学 | Method for breeding calanus finmarchicus with porcelain jar |
| CN104782577A (en) * | 2015-04-13 | 2015-07-22 | 临沂大学 | Method for breeding penilia avirostris with suction flasks |
| CN104982241A (en) * | 2015-08-04 | 2015-10-21 | 华中农业大学 | Method for evaluating safety of aquatic environment for genetically modified insect resistant rice |
| CN105891436A (en) * | 2016-06-30 | 2016-08-24 | 福建医科大学 | Biological testing method for comprehensively evaluating water quality of drinking water |
| CN107568165A (en) * | 2017-08-31 | 2018-01-12 | 环境保护部南京环境科学研究所 | A kind of experimental method evaluated genetically modified plants and influenceed on snail |
| CN111830223A (en) * | 2020-07-13 | 2020-10-27 | 南华大学 | Method for evaluating pollution level of uranium by utilizing daphnia magna |
| CN114431179A (en) * | 2022-02-25 | 2022-05-06 | 生态环境部南京环境科学研究所 | Evaluation method for safety influence of Bt insecticidal protein on daphnia magna |
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| CN101731185A (en) * | 2009-12-23 | 2010-06-16 | 大连水产学院 | Large-scale cultivation method for marine floating harpacticoida |
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Patent Citations (1)
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| CN101731185A (en) * | 2009-12-23 | 2010-06-16 | 大连水产学院 | Large-scale cultivation method for marine floating harpacticoida |
Non-Patent Citations (2)
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| 徐瑞薇等: "杀虫双环境影响和降解研究", 《环境科学丛刊》 * |
| 赵彩云等: "转Bt基因水稻对小型脊椎动物影响的研究概况", 《中国生物防治》 * |
Cited By (9)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104782576A (en) * | 2015-04-13 | 2015-07-22 | 临沂大学 | Method for breeding calanus finmarchicus with porcelain jar |
| CN104782577A (en) * | 2015-04-13 | 2015-07-22 | 临沂大学 | Method for breeding penilia avirostris with suction flasks |
| CN104782577B (en) * | 2015-04-13 | 2018-07-27 | 临沂大学 | A method of with bottle,suction culture beak tip Magna |
| CN104982241A (en) * | 2015-08-04 | 2015-10-21 | 华中农业大学 | Method for evaluating safety of aquatic environment for genetically modified insect resistant rice |
| CN104982241B (en) * | 2015-08-04 | 2017-10-24 | 华中农业大学 | A kind of method for evaluating transgenic pest-resistant rice aquatic environment security |
| CN105891436A (en) * | 2016-06-30 | 2016-08-24 | 福建医科大学 | Biological testing method for comprehensively evaluating water quality of drinking water |
| CN107568165A (en) * | 2017-08-31 | 2018-01-12 | 环境保护部南京环境科学研究所 | A kind of experimental method evaluated genetically modified plants and influenceed on snail |
| CN111830223A (en) * | 2020-07-13 | 2020-10-27 | 南华大学 | Method for evaluating pollution level of uranium by utilizing daphnia magna |
| CN114431179A (en) * | 2022-02-25 | 2022-05-06 | 生态环境部南京环境科学研究所 | Evaluation method for safety influence of Bt insecticidal protein on daphnia magna |
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