CN102498874B - Method for cultivating high folic acid enrichment spinach to strengthen folic acid nutrition of human bodies - Google Patents
Method for cultivating high folic acid enrichment spinach to strengthen folic acid nutrition of human bodies Download PDFInfo
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- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 title claims abstract description 153
- 241000219315 Spinacia Species 0.000 title claims abstract description 83
- 235000009337 Spinacia oleracea Nutrition 0.000 title claims abstract description 83
- 235000019152 folic acid Nutrition 0.000 title claims abstract description 79
- 239000011724 folic acid Substances 0.000 title claims abstract description 79
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 title claims abstract description 74
- 229960000304 folic acid Drugs 0.000 title claims abstract description 74
- 238000000034 method Methods 0.000 title claims abstract description 28
- 235000016709 nutrition Nutrition 0.000 title claims abstract description 15
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- 239000011159 matrix material Substances 0.000 claims abstract description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 16
- 239000012153 distilled water Substances 0.000 claims abstract description 13
- 239000012531 culture fluid Substances 0.000 claims description 17
- 238000009395 breeding Methods 0.000 claims description 12
- 238000005286 illumination Methods 0.000 claims description 12
- 230000001488 breeding effect Effects 0.000 claims description 7
- 239000012530 fluid Substances 0.000 claims description 7
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- 230000001105 regulatory effect Effects 0.000 claims description 4
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- 235000019354 vermiculite Nutrition 0.000 claims description 4
- 229910052902 vermiculite Inorganic materials 0.000 claims description 4
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- 206010016880 Folate deficiency Diseases 0.000 description 3
- 208000010188 Folic Acid Deficiency Diseases 0.000 description 3
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 description 3
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- MSTNYGQPCMXVAQ-KIYNQFGBSA-N 5,6,7,8-tetrahydrofolic acid Chemical compound N1C=2C(=O)NC(N)=NC=2NCC1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 MSTNYGQPCMXVAQ-KIYNQFGBSA-N 0.000 description 1
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- Cultivation Of Plants (AREA)
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Abstract
The invention discloses a method for cultivating high folic acid enrichment spinach to strengthen folic acid nutrition of human bodies, which comprises the following steps that: 1) a spinach accession TOT7337-B is determined as a high folic acid enrichment spinach; 2) bud forcing treatment is performed on the TOT7337-B seeds after being sterilized; 3) the TOT7337-B seeds treated by bud forcing are planted on a culture matrix to be cultivated through distilled water, the cultivation condition is 12 hours of light cultivation every day and 12 hours of dark cultivation, the cultivation on the culture matrix is stopped when the spinach grows to the height of 1.5 to 2.5cm, so the spinach seedling is obtained; and 4) the spinach seedling is transplanted in spinach culture liquid to be cultivated, the cultivation condition is 12 hours of light cultivation every day and 12 hours of dark cultivation every day, and the cultivation time is 4.5 to 5.5 weeks. Due to the adoption of the method, the spinach with high content of folic acid can b e produced within a short period of time.
Description
Technical field
Thereby the present invention relates to a kind of method of cultivating the high enrichment spinach of folic acid body folic acid nutrition.
Background technology
Folic acid refers to tetrahydrofolic acid and this class material of derivative thereof, belongs to water-soluble B family vitamin, and as main one carbon unit donor, folic acid participates in the biosynthesis of nucleic acid, amino acid, protein.Self can synthesize folic acid plant and microorganism, but human and other higher mammals lack complete folic acid synthetic system, can only from meals (mainly being that vegetable food comprises cereal and vegetables), absorb folic acid, and in the cereals, such as wheat, folate contents such as corn are very low, can not satisfy human body and recommend folic acid intake every day (adult: 400 μ g/ days, pregnant woman: 600 μ g/ days), leaf vegetables such as spinach contain higher folic acid in the greengrocery food, are the good sources of folic acid.Folic acid intake deficiency can cause diseases such as neonate's NTD, inborn defect, intelligence degeneration, megaloblastic anemia, and increases the risk of suffering from angiocardiopathy and cancer.The epidemiology evidence shows that the folic acid intake of a lot of populations all is being lower than recommendation folic acid intake in the world.China folic acid deficiency crowd is 2.6 hundred million, and annual nearly 1,800 ten thousand pregnant womens cause neonate's NTD (TNDs) because of afoliate acid.The northern neonate's NTD incidence of China is that the world is the highest.In the population that folic acid take in to lack, also comprise all very abundant developed countries of size of food and kind.Therefore, folic acid deficiency be one not only interfereed with the development in country but also influence the global health problem of developed country.
The method that improves human body folic acid intake at present comprises that taking in folic acid medicament, the reinforcement of food folic acid and folic acid biological strengthens.Although synthetic folic acid and the absorption folic acid medicament of strengthened artificial become the quick method of improving folic acid human intaking amount deficiency in food at present, its cost is higher, is difficult to promote in the developing country of poor relatively and remote districts.And the biological reinforced method that effectively solves global folic acid deficiency problem as another kind is advocated.Biological reinforced folic acid mainly is to realize folic acid biological reinforcement aspect by agricultural breeding technology screening homofolic acid vegetable variety or the transformation by the folic acid metabolism approach.Spinach utilizes the method for folic acid enrichment spinach body folic acid nutrition significant as a kind of vegetables commonly used.
Summary of the invention
Thereby the technical problem to be solved in the present invention provides a kind of method of cultivating the high enrichment spinach of folic acid body folic acid nutrition, the method has overcome the problems referred to above of conventional oral artificial synthetic folic acid medicament body folic acid nutrition, use this kind method and can produce the spinach with homofolic acid content between short-term, and cost is low, broad covered area.
In order solving the problems of the technologies described above, thereby to the invention provides a kind of method of cultivating the high enrichment spinach of folic acid body folic acid nutrition, to may further comprise the steps:
1), determines that TOT7337-B is as the high enrichment spinach of folic acid;
2), handle carrying out vernalization after the TOT7337-B seed disinfection;
3), on culture matrix, utilize distilled water to cultivate the TOT7337-B planting seed after the vernalization processing, breeding condition is: the illumination cultivation of 12 hours every days and 12 hours dark cultivation, temperature was 22 ± 0.5 ℃ when illumination was cultivated, temperature is 18 ± 0.5 ℃ during dark the cultivation, and light intensity is 700～800 μ mol m
-2s
-1Treat that spinach growth stops at the cultivation on the culture matrix when extremely highly being 1.5～2.5cm, get the spinach seedling;
4), the spinach seedling is transplanted in the spinach culture fluid cultivates, breeding condition is: the illumination of 12 hours every days is cultivated and 12 hours dark cultivation, and temperature is 22 ± 0.5 ℃ during the illumination cultivation, and temperature is 18 ± 0.5 ℃ when secretly cultivating, and light intensity is 700～800 μ mol m
-2s
-1The cultivation time was 4.5～5.5 weeks.
Thereby the improvement as the method for the high enrichment spinach of cultivation folic acid of the present invention body folic acid nutrition: culture matrix is that perlite, peat and vermiculite mix according to 1: 0.9～1.1: 0.9～1.1 volume ratio and get.
Thereby the further improvements in methods as the high enrichment spinach of cultivation folic acid of the present invention body folic acid nutrition: the preparation method of spinach culture fluid is:
By 1.43mM NH
4NO
3, 1.00mM CaCl
2, 1.64mM MgSO
4, 0.32mM NaH
2PO
4, 1.32mM K
2SO
4, 5.0 μ M MnCl
2, 0.2 μ M CuSO
4, 0.075 μ m (NH
4)
6Mo
7O
24, 1.90 μ M H
3BO
3, 20.0 μ M Fe-HEDTA, 1.0 μ M ZnSO
4, 0.1 μ M NiSO
4With surplus be that distilled water is formed the culture fluid base fluid; Regulating the pH value of cultivating the culture fluid base fluid again is 5.2～5.8.
Regulating the pH value is the conventional method of the industry, generally can utilize 0.1mol/L HCL and 0.1mol/L NaOH to carry out.
Thereby the further improvements in methods as the high enrichment spinach of cultivation folic acid of the present invention body folic acid nutrition: the cultivation of step 3) and step 4) is all carried out in cultivating the cave dish, cultivates the cave dish and is made by black plastic, and every cave specification is 15 * 15 * 15cm.
The present invention at first fills up culture matrix in every cave in step 3), and then place the TOT7337-B seed after 18～22 above-mentioned vernalization is handled; The cave dish is positioned in the blue frame of plastic to be cultivated; Cultivate with distilled water during this time, the consumption of distilled water is every cave 80～120ml every day; The present invention is in step 4), and every day, every cave added spinach culture fluid 80～120ml.
The present invention at first provides a kind of spinach kind TOT7337-B of superpower enrichment folic acid, and this spinach kind is to collect from us to whole world relevant unit to carry out a large amount of Screening and Identification nearly more than 70 kinds of spinach kinds and obtain; Primary Study shows that this variety resistance is stronger, and output and conventional variety are close, and has the ability of superpower enrichment folic acid.
Specific cultivation cave dish, culture matrix and spinach culture fluid are provided in breeding method of the present invention.
The spinach TOT7337-B that adopts the inventive method to cultivate and get, overground part folic acid enrichment concentration reaches the bright sample of 173 μ g/100g, with the content of folic acid transgene tomato is suitable in the world at present.Eat the bright spinach of about 400g every day, can satisfy human body and recommend folic acid intake requirement every day.And the spinach TOT7337-B that adopts conventional method to cultivate, its overground part folic acid enrichment concentration only is the bright samples of 118 μ g/100g.
Thereby the method for the high enrichment spinach of cultivation folic acid of the present invention body folic acid nutrition has following characteristics:
(1) spinach of the present invention's generation has homofolic acid content.
(2) the spinach cycle of the present invention's generation is short.
(3) spinach that produces of the present invention to compare artificial synthetic folic acid reagent cost low.
(4) utilize the approach of agricultural to improve the leaf of spinach acid content, do not change crowd's diet structure, accepted by the crowd easily.
Description of drawings
Below in conjunction with accompanying drawing the specific embodiment of the present invention is described in further detail.
Fig. 1 is gained spinach of the present invention and total folate content of commercially available conventional spinach and the comparison diagram of different folic acid derivatives content thereof;
Fig. 2 is the inventive method and total folate content of greenhouse gardening method and the comparison diagram of different folic acid derivatives content thereof.
Embodiment
Thereby embodiment 1, a kind of method of cultivating the high enrichment spinach of folic acid body folic acid nutrition are carried out following steps successively:
1), determine that TOT7337-B is as the high enrichment spinach of folic acid:
Screening and Identification obtains the high enrichment spinach of folic acid kind from a large amount of spinach germplasms of a large amount of global collections, and the spinach kind code name of the strongest enrichment folic acid is (TOT7337-B).
2), handle carrying out vernalization after the TOT7337-B seed disinfection:
Prepare 0.07% (volumetric concentration) liquor natrii hypochloritis earlier: get 0.7ml NaOCl and use the distilled water constant volume in the 1000ml volumetric flask.
After the TOT7337-B seed washes, put into above-mentioned 0.07% (volumetric concentration) liquor natrii hypochloritis and soaked 30 minutes, after water washes down, put into distilled water and soak vernalization 2 days.During this time, changed water once in per 24 hours.
3), first preparation culture matrix, this culture matrix is that perlite, peat and vermiculite mix according to 1: 1: 1 volume ratio and get.The particle diameter of perlite, peat and vermiculite is all controlled at 1～5mm.
Coil (15 * 15 * 15cm) in the cultivation cave that is filled with culture matrix the TOT7337-B planting seed after the vernalization processing; This cultivation cave dish is made by black plastic, and specification is every cave 15 * 15 * 15cm.Be specially: fill full culture matrix in every cave and (even culture matrix fills up 15 * 15 * 15cm), put into 20 in TOT7337-B seed after above-mentioned vernalization is handled in every cave; Last whole cave dish is positioned in the blue frame of plastic to be cultivated; Cultivate with distilled water during this time, the amount of distilled water is every cave 100ml every day; Breeding condition is: the illumination cultivation of 12 hours every days and 12 hours dark cultivation, and temperature was 22 ± 0.5 ℃ when illumination was cultivated, and secretly cultivating temperature is 18 ± 0.5 ℃, and light intensity is 700～800 μ mol m
-2s
-1After 7 days, spinach growth stops at the cultivation on the culture matrix when highly being about 2cm, gets the spinach seedling.
4), prepare the spinach culture fluid earlier:
The culture fluid base fluid is by 1.43mM NH
4NO
3, 1.00mM CaCl
2, 1.64mM MgSO
4, 0.32mM NaH
2PO
4, 1.32mM K
2SO
4, 5.0 μ M MnCl
2, 0.2 μ M CuSO
4, 0.075 μ M (NH
4)
6Mo
7O
24, 1.90 μ M H
3BO
3, 20.0 μ M Fe-HEDTA, 1.0 μ M ZnSO
4, 0.1 μ M NiSO
4With surplus be that distilled water is formed.
That is the NH that, contains 1.43mM in every liter of culture fluid base fluid
4NO
3, 1.00mM CaCl
2, 1.64mM MgSO
4, 0.32mM NaH
2PO
4, 1.32mM K
2SO
4, 5.0 μ M MnCl
2, 0.2 μ M CuSO
4, 0.075 μ M (NH
4)
6Mo
7O
24, 1.90 μ M H
3BO
3, the Fe-HEDTA of 20.0 μ M, the ZnSO of 1.0 μ M
4NiSO with 0.1 μ M
4, all the other are distilled water.It is 5.5 that above-mentioned culture fluid base fluid is regulated the pH value with 0.1mol/LHCl and 0.1mol/LNaOH, gets the spinach culture fluid.
The spinach seedling of step 3) gained is transplanted in the spinach culture fluid cultivates; Being about to the spinach culture fluid directly adds in the dish of cultivation cave, addition is according to every cave 100ml every day, the cultivation breeding condition is: the illumination cultivation of 12 hours every days and 12 hours dark cultivation, temperature was 22 ± 0.5 ℃ when illumination was cultivated, dark cultivation temperature is 18 ± 0.5 ℃, and light intensity is 700～800 μ mol m
-2s
-1The cultivation time was 5 weeks.Results.
Total folic acid and derivative content thereof compare test in experiment 1, the spinach sample (taking out large-leaved variety evening) buied with regard to the embodiment of the invention 1 gained spinach and local market, and example illustrates:
Its name of the spinach sample that market is buied is called the late Da Ye of taking out, be that the peasant utilizes common agriculture breeding method cultivation to obtain, the spinach of taking out Da Ye spinach fresh sample and gained of the present invention evening that we directly buy market sale compares (Fig. 1), and result of the test as shown in Figure 1.Total folic acid of spinach kind and derivative content thereof that gained spinach of the present invention and local market are buied find that relatively total folate content of gained spinach kind of the present invention and derivative content thereof all are higher than market spinach kind.
Test 2. couples of spinach kind TOT7337-B that the present invention relates to and other 26 kinds of spinach kinds collecting from other places, the world and carry out cultivation under the same terms, namely breeding method is the spinach breeding method (except spinach kind difference) that the embodiment of the invention 1 relates to.After waiting to gather in the crops, measure folic acid total amount in the spinach sample, result of the test is relatively found (table 1), under identical cultural method, namely under the spinach breeding method that the present invention relates to, kind TOT7337-B involved in the present invention contains higher folate content than other 26 kinds of spinach kinds, and namely its folic acid accumulation ability is far above other 26 kinds of kinds.
The different spinach kind of table 1. folic acid total content
| The kind numbering | Folic acid total content (mean value) |
| TOT7337-B | 173.21 |
| PI 261787 | 154.40 |
| NSL 6095 | 153.40 |
| PI 499372 | 148.34 |
| PI 200882 | 143.57 |
| PI 181809 | 143.10 |
| NSL 6085 | 142.79 |
| PI 285751 | 140.41 |
| NSL 92009 | 123.05 |
| PI 174384 | 121.04 |
| PI 175930 | 120.98 |
| PI 527332 | 114.58 |
| PI 174386 | 114.19 |
| PI 531456 | 113.77 |
| NSL 22149 | 110.96 |
| PI 174388 | 108.95 |
| PI 165710 | 107.41 |
| PI 321020 | 97.54 |
| PI 224959 | 96.85 |
| PI 263873 | 96.19 |
| PI 491262 | 95.85 |
| PI 179589 | 90.14 |
| PI 220546 | 89.70 |
| PI 433210 | 89.01 |
| TOT7337-A | 66.43 |
| TOT7339-B | 64.97 |
| PI 604792 | 54.05 |
The TOT7337-B seed that experiment 3. just relates among the present invention according to the method for the embodiment of the invention 1 cultivate with the spinach sample that obtains according to conventional greenhouse gardening spinach method in total folic acid and derivative content thereof compare test, example illustrates:
With embodiment 1 step 2) TOT7337-B seed after the vernalization of gained is handled plants in booth according to the spinach planting patterns of routine, and keeping the temperature in the booth is 30 ℃; Fertilising and water management were cultivated 42 days altogether according to general greenhouse gardening method; Results.After testing, the total folic acid in the booth gained spinach and derivative content thereof all are lower than the spinach (see figure 2) that the inventive method obtains.
At last, it is also to be noted that what more than enumerate only is several specific embodiments of the present invention.Obviously, the invention is not restricted to above embodiment, many distortion can also be arranged.All distortion that those of ordinary skill in the art can directly derive or associate from content disclosed by the invention all should be thought protection scope of the present invention.
Claims (2)
1. thereby cultivate the method for the high enrichment spinach of folic acid body folic acid nutrition, it is characterized in that may further comprise the steps:
1), determines that TOT7337-B is as the high enrichment spinach of folic acid;
2), handle carrying out vernalization after the TOT7337-B seed disinfection;
3), on culture matrix, utilize distilled water to cultivate the TOT7337-B planting seed after the vernalization processing, breeding condition is: the illumination cultivation of 12 hours every days and 12 hours dark cultivation, temperature was 22 ± 0.5 ℃ when illumination was cultivated, and light intensity is 700 ~ 800 μ mol m
-2s
-1, temperature is 18 ± 0.5 ℃ when secretly cultivating, and treats that spinach growth stops at the cultivation on the culture matrix when extremely highly being 1.5 ~ 2.5cm, get the spinach seedling;
To be perlite, peat and vermiculite according to the volume ratio of 1:0.9 ~ 1.1:0.9 ~ 1.1 mix described culture matrix gets;
4), the spinach seedling is transplanted in the spinach culture fluid cultivates, breeding condition is: the illumination of 12 hours every days is cultivated and 12 hours dark cultivation, and temperature is 22 ± 0.5 ℃ during the illumination cultivation, and light intensity is 700 ~ 800 μ mol m
-2s
-1, temperature is 18 ± 0.5 ℃ when secretly cultivating, the cultivation time was 4.5 ~ 5.5 weeks;
The preparation method of described spinach culture fluid is:
By 1.43 mM NH
4NO
3, 1.00 mM CaCl
2, 1.64 mM MgSO
4,
0.32 mM NaH
2PO
4, 1.32 mM K
2SO
4,
5.0 μ M MnCl
2, 0.2 μ M CuSO
4, 0.075 μ M (NH
4)
6Mo
7O
24, 1.90 μ M H
3BO
3, 20.0 μ M Fe-HEDTA, 1.0 μ M ZnSO
4, 0.1 μ M NiSO
4With surplus be that distilled water is formed the culture fluid base fluid; Regulating the pH value of cultivating the culture fluid base fluid again is 5.2 ~ 5.8.
2. thereby the method for the high enrichment spinach of cultivation folic acid according to claim 1 body folic acid nutrition, it is characterized in that: the cultivation of described step 3) and step 4) is all carried out in cultivating the cave dish, described cultivation cave dish is made by black plastic, and every cave specification is 15 * 15 * 15 cm.
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| JP2017143801A (en) * | 2016-02-18 | 2017-08-24 | 三菱ケミカル株式会社 | Plant cultivation method |
| CN106576782A (en) * | 2016-11-30 | 2017-04-26 | 蚌埠市众星蔬果科技专业合作社联合社 | Planting method for spinaches in summer |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101116401A (en) * | 2007-08-31 | 2008-02-06 | 浙江大学 | Method for planting iodine-contained spinach |
| CN101755557A (en) * | 2008-12-23 | 2010-06-30 | 北京玉佳明三态离子科学研究院有限公司 | Method for producing acoustic wave-fostered selenium-rich spinach |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101116401A (en) * | 2007-08-31 | 2008-02-06 | 浙江大学 | Method for planting iodine-contained spinach |
| CN101755557A (en) * | 2008-12-23 | 2010-06-30 | 北京玉佳明三态离子科学研究院有限公司 | Method for producing acoustic wave-fostered selenium-rich spinach |
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Application publication date: 20120620 Assignee: Zhejiang Green Ecological Agriculture Co., Ltd. Assignor: Zhejiang University Contract record no.: 2015330000234 Denomination of invention: Method for cultivating high folic acid enrichment spinach to strengthen folic acid nutrition of human bodies Granted publication date: 20130807 License type: Exclusive License Record date: 20150901 |
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