CN102007846A - Method for detecting quantity of nitrogen fixed by soybean root nodules - Google Patents
Method for detecting quantity of nitrogen fixed by soybean root nodules Download PDFInfo
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- CN102007846A CN102007846A CN 201010534835 CN201010534835A CN102007846A CN 102007846 A CN102007846 A CN 102007846A CN 201010534835 CN201010534835 CN 201010534835 CN 201010534835 A CN201010534835 A CN 201010534835A CN 102007846 A CN102007846 A CN 102007846A
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Abstract
The invention relates to a method for detecting the quantity of nitrogen fixed by soybean root nodules, which relates to a method for detecting the quantity of the nitrogen fixed by root nodules and solves the problem of low accuracy of the conventional method for detecting the nitrogen fixed by the soybean root nodules. The method comprises the following steps of: determining the variety of detected soybeans and the variety of homologous near-allele non-nodulating soybeans; soaking seeds of the detected soybeans and the homologous near-allele non-nodulating soybeans in aqueous solution of hydrogen peroxide (H2O2) and then sowing the seeds in culture pots respectively, wherein three strains are planted in each pot and cultured under the same growth condition; performing destructive sampling on the strains at the maturation stage; detecting the total nitrogen quantity of the detected soybeans and the homologous near-allele non-nodulating soybeans respectively; and subtracting the total nitrogen quantity of the homologous near-allele non-nodulating soybeans from that of the detected soybeans to obtain the quantity of the nitrogen fixed by root nodules of the detected soybeans. The method has the detection accuracy of over 90 percent and can be used for detecting the quantity of the nitrogen fixed by the root nodules of leguminous crops.
Description
Technical field
The present invention relates to the detection method of nodule nitrogen fixation amount.
Technical background
Soybean nodulation is the organ of soybean fixed nitrogen, and its major function is fixing N in the atmosphere
2Be converted into organic nitrogen, for nitrogen nutrition is provided greatly.The size of soybean nodulation nitrogen fixing capacity determines its fixedly ability of conforming of the nitrogenous source in the atmosphere, has also determined the growing power of soybean.The soybean nodulation nitrogen fixing capacity is said evaluation index as the core of soybean nodulation function and is become the importance of soybean physiological Study work.The detection method of existing soybean nodulation amount of nitrogen fixation has two kinds, and the first adopts not that nitrogen fertilizer application is treated to contrast, and it two is to adopt isotope
15The N labelled fertilizer utilizes minusing to calculate the nodule nitrogen fixation amount for for examination fertilizer, and these two kinds of methods disturbing factor in measuring process is more, error is big, and partial results is out in the cold, causes the nodule nitrogen fixation total amount on the low side, make detected nodule nitrogen fixation amount data precision in 60%～70% scope, low precision.
Summary of the invention
The present invention is the problem for the low precision of the detection method that solves existing soybean nodulation amount of nitrogen fixation, and the detection method of soybean nodulation amount of nitrogen fixation is provided.
The detection method of soybean nodulation amount of nitrogen fixation of the present invention, carry out according to the following steps: one, at first determine soybean varieties, choose the close and/or maternal close non-dross soybean line of male parent, from the non-dross soybean line of having chosen, choose again and plant the more approaching strain of physiological trait with tested kind strain, measure gene order by the molecular biology ment then, filter out and the immediate soybean line of tested kind non-dross soybean line in contrast, obtain the nearly allelomorph non-nodulating of the homology soybean varieties of tested soybean varieties; Two, the seed mass concentration of the nearly allelomorph non-nodulating of the homology soybean varieties of the tested soybean varieties that the tested soybean varieties and the step 1 of step 1 are selected is 1.8%～2.3% H
2O
2Aqueous solution soaking 0.5h～1h cleans with clear water then; Three, the soil sieve in 5mm aperture, fine earth after sieving is packed in the clean Culture basin, every basin soil loading amount is 10kg～12kg, apply 0.7g～1.0g urea simultaneously, 0.3g～0.5g diammonium hydrogen phosphate, 0.2g～0.4g potassium sulphate, to broadcast respectively in the different Culture basins through the seed of the nearly allelomorph non-nodulating of the homology soybean varieties of the seed of the tested soybean varieties that step 2 is handled and tested soybean then, every basin is broadcast 5 seeds, final singling 3 strains are cultivated under identical growth conditions after emerging; Four, carry out the destructiveness sampling in the maturing stage of soybean, be divided into overground part and underground part two parts, the nitrogen pool to the nearly allelomorph non-nodulating of the homology of tested soybean varieties and tested soybean kind detects respectively; Five, deduct the nitrogen pool of the nearly allelomorph non-nodulating of the homology kind of tested soybean with the nitrogen pool of tested soybean varieties, obtain tested soybean nodulation amount of nitrogen fixation.
The present invention adopts the test material of the nearly allelomorph non-nodulating of homology kind for contrast, nodule nitrogen fixation process and plant absorbing nitrogen process combine together, make result of the test more approach reality, make detected nodule nitrogen fixation amount data accurate, accuracy is more than 90%, than its existing employing not nitrogen fertilizer application be treated to contrast or adopt isotope
15The N labelled fertilizer is for improving 15%～18% for the poor then method precision that subtracts of examination fertilizer, and detection efficiency improves 2 times～3 times.This method can be used for the mensuration of the nodule nitrogen fixation amount of legume.
Embodiment
Embodiment one: the detection method of the soybean nodulation amount of nitrogen fixation of present embodiment, carry out according to the following steps: one, at first determine soybean varieties, choose the close and/or maternal close non-dross soybean line of male parent, from the non-dross soybean line of having chosen, choose again and plant the more approaching strain of physiological trait with tested kind strain, measure gene order by the molecular biology ment then, filter out and the immediate soybean line of tested kind non-dross soybean line in contrast, obtain the nearly allelomorph non-nodulating of the homology soybean varieties of tested soybean varieties; Two, the seed mass concentration of the nearly allelomorph non-nodulating of the homology soybean varieties of the tested soybean varieties that the tested soybean varieties and the step 1 of step 1 are selected is 1.8%～2.3% H
2O
2Aqueous solution soaking 0.5h～1h cleans with clear water then; Three, the soil sieve in 5mm aperture, fine earth after sieving is packed in the clean Culture basin, every basin soil loading amount is 10kg～12kg, apply 0.7g～1.0g urea simultaneously, 0.3g～0.5g diammonium hydrogen phosphate, 0.2g～0.4g potassium sulphate, to broadcast respectively in the different Culture basins through the seed of the nearly allelomorph non-nodulating of the homology soybean varieties of the seed of the tested soybean varieties that step 2 is handled and tested soybean then, every basin is broadcast 5 seeds, final singling 3 strains are cultivated under identical growth conditions after emerging; Four, carry out the destructiveness sampling in the maturing stage of soybean, be divided into overground part and underground part two parts, the nitrogen pool to the nearly allelomorph non-nodulating of the homology of tested soybean varieties and tested soybean kind detects respectively; Five, deduct the nitrogen pool of the nearly allelomorph non-nodulating of the homology kind of tested soybean with the nitrogen pool of tested soybean varieties, obtain tested soybean nodulation amount of nitrogen fixation.
The present invention adopts the test material of the nearly allelomorph non-nodulating of homology kind for contrast, nodule nitrogen fixation process and plant absorbing nitrogen process combine together, make result of the test more approach reality, make detected nodule nitrogen fixation amount data accurate, accuracy is more than 90%, than its existing employing not nitrogen fertilizer application be treated to contrast or adopt isotope
15The N labelled fertilizer is for improving 15%～18% for the poor then method precision that subtracts of examination fertilizer, and detection efficiency improves 2 times～3 times.
Embodiment two: what present embodiment and embodiment one were different is: H in the step 2
2O
2The mass concentration of the aqueous solution is 1.9%～2.2%, and soak time is 0.6h～0.9h.Other is identical with embodiment one.
Embodiment three: what present embodiment and embodiment one were different is: H in the step 2
2O
2The mass concentration of the aqueous solution is 2.0%, and soak time is 0.8h.Other is identical with embodiment one.
Embodiment four: what present embodiment was different with one of embodiment one to three is: the soil loading amount of each Culture basin is 10.5kg～11.5kg in the step 3, apply 0.75g～0.95g urea simultaneously, 0.35g～0.45g diammonium hydrogen phosphate, 0.25g～0.35g potassium sulphate.Other is identical with one of embodiment one to three.
Embodiment five: what present embodiment was different with one of embodiment one to three is: the soil loading amount of each Culture basin is 11.2kg in the step 3, applies 0.83g urea simultaneously, 0.42g diammonium hydrogen phosphate, 0.42g potassium sulphate.Other is identical with one of embodiment one to three.
Embodiment six: the detection method of the soybean nodulation amount of nitrogen fixation of present embodiment, carry out according to the following steps: one, determine that at first soybean varieties is Clark 3, choose close with its male parent and maternal close non-dross soybean line, from the non-dross soybean line of having chosen, choose again and plant the more approaching strain of physiological trait with tested kind strain, measure gene order by the molecular biology ment at last, filter out at last and the immediate soybean line of tested kind non-dross soybean line in contrast, obtain the nearly allelomorph non-nodulating of the homology kind soybean Harosoy 37 of tested soybean; Two, the seed mass concentration of the soybean Harosoy 37 that the tested soybean Clark 3 and the step 1 of step 1 are selected is 2.0% H
2O
2Aqueous solution soaking 0.8h cleans with clear water then; Three, the soil sieve in 5mm aperture, fine earth after sieving is packed in the clean Culture basin, every basin soil loading amount is 10kg, apply 0.80g urea simultaneously, 0.40g diammonium hydrogen phosphate, 0.30g potassium sulphate, to broadcast respectively in the different Culture basins through the seed of the soybean Clark 3 that step 2 is handled and the seed of soybean Harosoy 37 then, every basin is broadcast 5 seeds, and back final singling 3 strains of emerging are cultivated under identical growth conditions; Four, carry out the destructiveness sampling in the maturing stage of crop, the nitrogen pool to soybean Clark 3 and soybean Harosoy 37 detects respectively; Five, deduct the nitrogen pool of soybean Harosoy 37 with the nitrogen pool of soybean Clark 3, obtain the nodule nitrogen fixation amount of soybean Clark 3.
The nitrogen pool of tested legume kind soybean Clark 3 is the 83.8mg/ strain in the present embodiment step 4, and the nitrogen pool of soybean Harosoy 37 is the 53.4mg/ strain, thereby the nodule nitrogen fixation amount of soybean Clark 3 is the 30.4mg/ strain.
Present embodiment adopts the test material of the nearly allelomorph non-nodulating of homology kind for contrast, nodule nitrogen fixation process and plant absorbing nitrogen process combine together, make result of the test more approach reality, make detected nodule nitrogen fixation amount data accurate, accuracy is more than 90%, than its existing employing not nitrogen fertilizer application be treated to contrast or adopt isotope
15The N labelled fertilizer is not considered plantation and incubation for improving 15.3% for the poor then method precision that subtracts of examination fertilizer, and detecting each sample average only needs 10～15 minutes, and existing conventional detection needs about 30 minutes, and detection efficiency improves 2～3 times.
Claims (5)
1. the detection method of soybean nodulation amount of nitrogen fixation, the detection method that it is characterized in that the soybean nodulation amount of nitrogen fixation is carried out according to the following steps: one, at first determine soybean varieties, choose the close and/or maternal close non-dross soybean line of male parent, from the non-dross soybean line of having chosen, choose again and plant the more approaching strain of physiological trait with tested kind strain, measure gene order by the molecular biology ment then, filter out and the immediate soybean line of tested kind non-dross soybean line in contrast, obtain the nearly allelomorph non-nodulating of the homology soybean varieties of tested soybean varieties; Two, the seed mass concentration of the nearly allelomorph non-nodulating of the homology soybean varieties of the tested soybean varieties that the tested soybean varieties and the step 1 of step 1 are selected is 1.8%～2.3% H
2O
2Aqueous solution soaking 0.5h～1h cleans with clear water then; Three, the soil sieve in 5mm aperture, fine earth after sieving is packed in the clean Culture basin, every basin soil loading amount is 10kg～12kg, apply 0.7g～1.0g urea simultaneously, 0.3g～0.5g diammonium hydrogen phosphate, 0.2g～0.4g potassium sulphate, to broadcast respectively in the different Culture basins through the seed of the nearly allelomorph non-nodulating of the homology soybean varieties of the seed of the tested soybean varieties that step 2 is handled and tested soybean then, every basin is broadcast 5 seeds, final singling 3 strains are cultivated under identical growth conditions after emerging; Four, carry out the destructiveness sampling in the maturing stage of soybean, be divided into overground part and underground part two parts, the nitrogen pool to the nearly allelomorph non-nodulating of the homology of tested soybean varieties and tested soybean kind detects respectively; Five, deduct the nitrogen pool of the nearly allelomorph non-nodulating of the homology kind of tested soybean with the nitrogen pool of tested soybean varieties, obtain tested soybean nodulation amount of nitrogen fixation.
2. the detection method of soybean nodulation amount of nitrogen fixation according to claim 1 is characterized in that H in the step 2
2O
2The mass concentration of the aqueous solution is 1.9%～2.2%, and soak time is 0.6h～0.9h.
3. the detection method of soybean nodulation amount of nitrogen fixation according to claim 1 is characterized in that H in the step 2
2O
2The mass concentration of the aqueous solution is 2.0%, and soak time is 0.8h.
4. according to the detection method of claim 1,2 or 3 described soybean nodulation amount of nitrogen fixation, the soil loading amount that it is characterized in that each Culture basin in the step 3 is 10.5kg～11.5kg, apply 0.75g～0.95g urea simultaneously, 0.35g～0.45g diammonium hydrogen phosphate, 0.25g～0.35g potassium sulphate.
5. according to the detection method of claim 1,2 or 3 described soybean nodulation amount of nitrogen fixation, the soil loading amount that it is characterized in that each Culture basin in the step 3 is 11.2kg, applies 0.83g urea simultaneously, 0.42g diammonium hydrogen phosphate, 0.42g potassium sulphate.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103891506A (en) * | 2014-03-28 | 2014-07-02 | 中国科学院东北地理与农业生态研究所 | Measuring and calculating method of field soybean root nodule nitrogen fixation amount |
| CN106226331A (en) * | 2016-07-18 | 2016-12-14 | 辽宁石油化工大学 | A kind of method measuring bulk soil soil denitrification |
| CN106556667A (en) * | 2016-11-25 | 2017-04-05 | 辽宁石油化工大学 | A kind of assay method of rhizosphere area soil nitrogen fixation |
| CN106556688A (en) * | 2016-11-25 | 2017-04-05 | 辽宁石油化工大学 | A kind of assay method of rhizosphere area soil ammonification |
| CN106771054A (en) * | 2016-11-25 | 2017-05-31 | 辽宁石油化工大学 | A kind of method of measure bulk soil soil cellulysis |
-
2010
- 2010-11-08 CN CN2010105348357A patent/CN102007846B/en not_active Expired - Fee Related
Non-Patent Citations (4)
| Title |
|---|
| 《农业系统科学与综合研究》 20100831 苗淑杰等 结瘤和非结瘤近等位基因品系大豆地上部生长性状 317-320 1-5 第26卷, 第3期 2 * |
| 《大豆科学》 20100430 苗淑杰等 大豆结瘤固氮的分子生理研究 319-324 1-5 第29卷, 第2期 2 * |
| 《微生物学通报》 19830630 贾凤菊 大田大豆有效根瘤固氮活力简易估测方法 246-248 1-5 , 第6期 2 * |
| 《韶关学院学报》 20090331 黎林 生物固氮的研究现状初探 88-91 1-5 第30卷, 第3期 2 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103891506A (en) * | 2014-03-28 | 2014-07-02 | 中国科学院东北地理与农业生态研究所 | Measuring and calculating method of field soybean root nodule nitrogen fixation amount |
| CN103891506B (en) * | 2014-03-28 | 2015-08-19 | 中国科学院东北地理与农业生态研究所 | The measuring method of field soybean nodule nitrogen fixation amount |
| CN106226331A (en) * | 2016-07-18 | 2016-12-14 | 辽宁石油化工大学 | A kind of method measuring bulk soil soil denitrification |
| CN106556667A (en) * | 2016-11-25 | 2017-04-05 | 辽宁石油化工大学 | A kind of assay method of rhizosphere area soil nitrogen fixation |
| CN106556688A (en) * | 2016-11-25 | 2017-04-05 | 辽宁石油化工大学 | A kind of assay method of rhizosphere area soil ammonification |
| CN106771054A (en) * | 2016-11-25 | 2017-05-31 | 辽宁石油化工大学 | A kind of method of measure bulk soil soil cellulysis |
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