CN113273417A - Application of boron fertilizer in inhibiting absorption, storage and cadmium transfer of winter wheat and inhibition method - Google Patents
Application of boron fertilizer in inhibiting absorption, storage and cadmium transfer of winter wheat and inhibition method Download PDFInfo
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Abstract
The invention provides an application of a boron fertilizer in inhibiting absorption, storage and transport of cadmium in winter wheat and an inhibiting method, and relates to the technical field of farmland pollution remediation. In the invention, the soil application of boron fertilizer before sowing winter wheat can obviously inhibit the absorption of cadmium in seedling stage and jointing stage of winter wheat: the reduction range of cadmium accumulation of winter wheat in the seedling stage is 43.90 percent; the reduction range of the cadmium content at the root and the overground part in the jointing stage is 52.11-74.59%. The leaf surface spraying of the boric fertilizer before the jointing stage can also obviously inhibit the cadmium absorption of the winter wheat, and the cadmium content reduction amplitude of the roots and the overground parts is 39.37-72.02 percent; compared with the method without applying boron, the method has the advantages that the method has the tendency of improving the cadmium content of different parts in the flowering period and the mature period by applying the boron fertilizer, and simultaneously, the cadmium content in soil is improved by applying the boron fertilizer. In a word, boron regulates the cadmium transport in the whole growth period of wheat by regulating the expression of the cadmium transport gene, and the effect of boron on inhibiting cadmium absorption in the seedling period is most obvious.
Description
Technical Field
The invention belongs to the technical field of farmland pollution remediation, and particularly relates to an application of boron fertilizer in inhibiting absorption, storage and cadmium transfer of winter wheat and an inhibiting method.
Background
Cadmium (Cd) has very high toxicity to wheat growth. Under normal conditions, except that some Cd hyperaccumulating plants can accumulate higher Cd content, most plants generate toxic symptoms when the concentration of Cd/g dry matter exceeds 5-10 mu g. In plants, the main and common symptoms of cadmium poisoning are yellowing, mesophyll atrophy and stunting. Other cadmium toxicities include altering chloroplast ultrastructure, inhibiting photosynthesis, inducing lipid peroxidation, inhibiting pollen germination and pollen tube growth, and cadmium has been shown to interfere with the uptake, transport and utilization of several elements and water, resulting in affected plant growth. The food code committee has set the safety threshold for cadmium content in grains to 0.4mg/kg rice (0.2 mg/kg in china), 0.2mg/kg wheat, 0.1mg/kg corn and barley, and a maximum allowable limit of 70 μ g per day for human cadmium in the joint office of the Food and Agriculture Organization (FAO) and the World Health Organization (WHO) and the FAO/WHO (food code committee, 2006). Therefore, controlling the accumulation of Cd in crop seeds is an important measure for protecting human health.
The transfer of cadmium from soil to grain involves uptake of the root system from the soil, transfer from root epidermal cells to xylem to above ground, and transport or reactivation to grain. For most plants, the roots accumulate higher levels of cadmium than other tissues. The roots have a series of mechanisms that prevent the accumulation of cadmium in the aerial parts, such as the production of cadmium chelates in the cytoplasm of the root cells, the retention of cadmium chelates in the vacuole, the restriction of the transfer of cadmium from the symplast to the xylem, and the formation of physical barriers that hinder the movement of cadmium out of the xylem cells, in order to restrict the transfer of cadmium from the apoplast to the xylem. Subcellular compositional analysis showed that the soluble components and cell walls had higher cadmium concentrations in the plant root system. Studies have shown that Cd transporters are involved in all these processes. The transporters OsNramp5, HvNramp5, OsIRT1, OsIRT2, OsNRAMP1 and OsCd1 are responsible for the transport of cadmium from soil solutions to rice and barley root cells, respectively. The transporter OsHMA3 is responsible for root vacuolar sequestration, and the transporters OsHMA2, OsHMA3, OsZIP7 and OsCAL1 play an important role in root-crown transfer of Cd. The transporters OsHMA2, OsLCT1, OsZIP7 and OsCCX2 are responsible for Cd partitioning to grain.
In addition, a variety strategy for reducing the absorption and accumulation of the cadmium in the wheat is also provided, and comprises the steps of externally applying a plant growth regulator, an inorganic modifier, properly applying fertilizer, silicon and organic fertilizer, biochar and the like. The nutrition management inhibits cadmium absorption by competing cadmium absorption sites, improves the wheat oxidation resistance system, enhances cadmium stress, and is an effective measure for reducing cadmium toxicity. Nitrogen (N), phosphorus (P) and potassium (K) play important roles in Cd stress tolerance. The elements are used as macroelements in plant nutrition, can promote plant growth, reduce activity of cadmium in soil, and are absorbed by platinum. Sulfur is important in sulfur-containing compounds, such as cysteine (Cys), methionine (Met), GSH, and several coenzymes that play important roles in cadmium uptake and tolerance. The trace elements such as calcium (Ca), magnesium (Mg), iron (Fe), manganese (Mn), copper (Cu) and zinc (Zn) can inhibit the absorption and accumulation of cadmium by plants. This may be associated with absorption of cadmium by some transport systems involved in micronutrient absorption. Meanwhile, beneficial elements such as selenium (Se), silicon (Si) and the like can improve root growth and photosynthetic parameters and reduce cadmium poisoning. These nutrients can increase plant biomass and yield, compete for the same membrane transporters, produce Phytochelatins (PCs), and avoid Cd accumulation in plants.
Boron (B) is an essential micronutrient and plays an important role in the high yield and quality of crops. In recent years, it has been found that B plays an important role in alleviating abiotic stresses in plants, including aluminum (Al) and cadmium poisoning. Studies have shown that boron mitigates cadmium accumulation and toxicity by inhibiting cadmium uptake and down regulating cadmium transport genes and improving oxidative stress. However, the boron application pattern and its effect on Cd uptake during the whole growth phase, especially the mature phase, of wheat is not clear.
Disclosure of Invention
In view of the above, the invention aims to provide an application of boron fertilizer in inhibiting absorption, storage and transport of cadmium in winter wheat and an inhibition method, which inhibit absorption, storage and transport of cadmium in the whole growth period of wheat by regulating expression of cadmium absorption, storage and transport genes and provide theoretical support for a mechanism of inhibiting cadmium absorption of wheat by boron.
In order to achieve the above object, the present invention provides the following technical solutions:
the invention provides an application of boron fertilizer in inhibiting absorption, storage and transport of cadmium in winter wheat.
Preferably, the boron fertilizer inhibits the absorption, storage and transport of cadmium by winter wheat in a seedling stage by regulating the expression of genes related to the absorption, storage and transport of cadmium of winter wheat.
Preferably, the genes associated with cadmium uptake include triee 5660, triee 5770, and TCONS 5200; the nucleotide sequence of the TRIAE5660 is shown as SEQ ID NO.1, and the nucleotide sequence of the TRIAE5770 is shown as SEQ ID NO. 2; the nucleotide sequence of the TCONS5200 is shown as SEQ ID NO. 6;
the related genes for storing cadmium comprise TCONS1113 and TRIAE 5370; the nucleotide sequence of the TCONS1113 is shown as SEQ ID NO. 3; the nucleotide sequence of the TRIAE5370 is shown as SEQ ID NO. 4;
genes associated with cadmium transport include triee 5770 and triee 1060; the nucleotide sequence of the TRIAE1060 is shown as SEQ ID NO. 5.
The invention provides a method for inhibiting absorption, storage and transport of cadmium in winter wheat, which comprises the following steps: before sowing winter wheat, applying boron fertilizer in the form of base fertilizer, or before jointing stage, spraying boron fertilizer on leaf surface.
Preferably, the cadmium content of the winter wheat planting field is 0-4 mg/kg.
Preferably, the boron fertilizer is H3BO3The total amount of the fertilizer applied to each mu is 1.0-1.5 kg.
Preferably, the application includes soil application and foliar spray application.
The invention provides an application of boron fertilizer in inhibiting absorption, storage and transport of cadmium in winter wheat. The embodiment of the invention adopts a pot experiment (the cadmium content in soil is 4mg/kg) to verify the influence of two application modes of soil application and leaf surface spraying on the biomass, the cadmium content and the cadmium transport gene expression quantity of winter wheat in different growth periods. The results show that the best fertilization mode is wheat sowingBefore planting, the fertilizer is applied to soil in the form of trace element nutrient solution, and the application amount of boron fertilizer is 300mL 46.2mmol/L H3BO3Nutrient solution (3 mL of said H per kg of soil)3BO3Nutrient solution), the final soil boron concentration is 1.5mg B/kg soil (calculated according to soil with 20cm of plough layer, the soil weight per mu is about 150000kg, and the application amount of the boron fertilizer is about 1.285kg H per mu3BO3) Can obviously inhibit the absorption of cadmium in the seedling stage and the jointing stage of the winter wheat: in the seedling stage, the accumulation of cadmium in winter wheat is obviously reduced by applying boron fertilizer in soil, and the reduction range is 43.90 percent; in the jointing stage, the cadmium content of roots and overground parts is obviously reduced by applying boron fertilizer on soil and spraying boron fertilizer on leaf surfaces; compared with the treatment without applying boron, the boron fertilizer application has the tendency of improving the cadmium content of different parts in the flowering period and the mature period, and simultaneously, the boron fertilizer application improves the cadmium content in soil, which shows that the cadmium absorption can be reduced and the transport to the overground part can be improved by spraying the boron fertilizer on the leaf surfaces; fluorescence quantitative PCR analysis showed that, the tiae 5660 and the tiae 5770 play an important role in cadmium uptake during the seedling stage. TCONS1113 and triee 5370 play an important role in vacuolar storage of cadmium during flowering, and triee 5770 and triee 1060 play an important role in cadmium migration to grain. In a word, boron regulates the cadmium transport in the whole growth period of wheat by regulating the expression of the cadmium transport gene, and the effect of boron on inhibiting cadmium absorption in the seedling period is most obvious.
Drawings
FIG. 1 is a plot of cadmium concentration at the seedling stage of wheat, where the data represent the mean of quadruplicate. + -. standard error, with different lower case letters indicating significant differences between treatments, (multiple range test for LSD, P <0.05), the following;
FIG. 2 is the concentration of cadmium in the roots and upper parts of the wheat during the jointing stage, wherein A represents the roots and B represents the upper parts of the ground;
FIG. 3 shows the Cd content in the roots, leaves and ears of wheat at the flowering stage, where A represents the roots, B represents the leaves, and C represents the ears;
FIG. 4 shows the Cd content in the wheat mature root, stem, hull and grain, wherein A represents the root. B represents stem, C represents shell, D represents kernel;
FIG. 5 shows the Cd content in soil at maturity;
FIG. 6 is a graph showing the mean value of three repetitions plus or minus standard error in a bar graph, and different lower case letters show that significant differences exist between different treatments, wherein the relative expression levels of 6 Cd transporter genes in a seedling stage are detected by qRT-PCR;
FIG. 7 is a graph showing the mean of six replicates plus or minus the standard error for the bar graph showing significant differences between different treatments using qRT-PCR technology to study the relative expression levels of 6 Cd transporter genes at flowering time;
figure 8 is the concentration of phosphorus, zinc, iron, manganese, copper and boron in wheat grain, wherein a represents phosphorus, B represents zinc, C represents iron, D represents manganese, E represents copper, F represents boron; and the bars represent the mean ± standard error of six replicates, with different lower case letters indicating significant differences between treatments.
Detailed Description
The invention provides an application of boron fertilizer in inhibiting absorption, storage and transport of cadmium in winter wheat.
The boron fertilizer preferably inhibits the absorption, storage and transport of cadmium by regulating the expression of related genes of the winter wheat and the cadmium absorption, storage and transport of cadmium in the seedling stage; the related genes of cadmium absorption comprise TRIAE5660, TRIAE5770 and TCONS 5200; the nucleotide sequence of the TRIAE5660 is shown as SEQ ID NO.1, and the nucleotide sequence of the TRIAE5770 is shown as SEQ ID NO. 2; the nucleotide sequence of the TCONS5200 is shown as SEQ ID NO. 6; the related genes for storing cadmium comprise TCONS1113 and TRIAE 5370; the nucleotide sequence of the TCONS1113 is shown as SEQ ID NO. 3; the nucleotide sequence of the TRIAE5370 is shown as SEQ ID NO. 4; genes associated with cadmium transport include triee 5770 and triee 1060; the nucleotide sequence of the TRIAE1060 is shown as SEQ ID NO. 5.
In the invention, the boron fertilizer can preferably inhibit the expression of the winter wheat root and the overground part TCONS1113, TRIAE5370, TRIAE5770 and TRIAE5660 before the jointing stage and promote the expression of the winter wheat root and the overground part TCONS5200 and TRIAE 1060. The specific type and source of the boron fertilizer are not particularly limited in the invention, and boric acid and other boron-containing substances are preferably included.
In the present invention, the above 6 genes are mainly expressed in the root, while the TRIAE5770 and TRIAE5660 are hardly detected in the leaf and ear of wheat at the flowering stage. Cd + B treated trioe 1060 and trioe 5770 had a tendency to increase the relative expression level in the root system compared to Cd treatment, while TCONS1113, TCONS5200, trioe 5370 and trioe 5660 decreased. And the genes except TRIAE5770 all increased with increasing boron concentration, especially in roots and leaves. Meanwhile, the cadmium concentration is negatively correlated with the expression of TCONS5200(P <0.01) TCONS1113, trie 5370 and triee 1060 in roots, with the expression of trie 5660, trie 5770 and TCONS1113 in leaves, and with the expression of trie 5370 and trie 1113 in ears. The cadmium concentration is significantly and positively correlated with TRIAE5770(P <0.01), TRIAE5660(P <0.01) and TRIAE1060(P < 0.05). Expression of these genes may play an important role in regulating Cd uptake and transport.
The invention provides a method for inhibiting absorption, storage and transport of cadmium in winter wheat, which comprises the following steps: before sowing winter wheat, applying boron fertilizer in the form of base fertilizer, or before jointing stage, spraying boron fertilizer on leaf surface.
The cadmium content of the winter wheat planting field is preferably 0-4 mg/kg. The boron fertilizer of the invention is H3BO3The total amount of the fertilizer applied per mu is preferably 1.0-1.5 kg. The method of application is not particularly limited in the present invention, and preferably includes soil application and foliar spray.
In the pot simulation experiment of the embodiment of the invention, the boron fertilizer is applied to winter wheat in different periods, so that different application effects in different periods are found, and the wheat can absorb cadmium in the early growth stage when no boron fertilizer exists; the Cd + B treatment was 43.90% lower (P <0.05) than Cd treatment. The result shows that the boron application can obviously reduce the absorption of Cd in the seedling stage of wheat. In the jointing stage of wheat, the Cd content in the root is higher than that in the overground part. The Cd contents in the roots treated by Cd + B, F0.1.1% (boric acid solution with the foliar spray mass concentration of 0.1%) and F0.3% (boric acid solution with the foliar spray mass concentration of 0.3%) are respectively reduced by 74.59% (P <0.05), 46.63% (P <0.05) and 72.02% (P <0.05) compared with the Cd treatment; overground Cd + B, F0.1.1%, F0.3% and F0.6% treatments were reduced by 52.11% (P <0.05), 39.37% (P <0.05), 47.46% (P <0.05) and 72.02% (P <0.05), respectively, over Cd treatment. Therefore, the Cd content of the wheat root and the overground part in the jointing stage is obviously reduced no matter the boron fertilizer is applied to soil or the boron fertilizer is applied to leaf surfaces. But in the flowering phase, the treatment rates of Cd + B, F0.3.3% and F0.6% in the root system are respectively improved by 174.55% (P <0.05), 38.76% and 413.46% (P <0.05) compared with the treatment rate of Cd, and the treatment rates of Cd + B, F0.1.1% and F0.6% in the leaf are respectively improved by 178.29% (P <0.05), 51.20% and 67.10% compared with the treatment rate of Cd; the Cd + B and F treatment of 0.6% in the grains is respectively improved by 106.1% (P is less than 0.05) and 15.55% compared with the Cd treatment. Therefore, the cadmium content of different parts of the wheat can be improved by applying the boron fertilizer in the flowering phase of the wheat, and the increase range of the boron application amount of the soil is larger than that of the boron spraying on the leaf surfaces. In the mature period, the concentrations of Cd treated by Cd + B, F0.1.1%, F0.3% and F0.6% in the root system are respectively increased by 90.38% (P <0.05), 58.72%, 49.59% and 107.5% (P <0.05) compared with Cd treatment; the concentration of Cd treated by Cd + B, F0.1.1%, F0.3% and F0.6% in the stem is respectively increased by 7.81%, 25.53%, 2.04% and 8.36% compared with that of Cd treatment; the concentrations of Cd treated by Cd + B, F0.1.1%, F0.3% and F0.6% in glumes are respectively reduced by 29.77%, 28.66%, 23.95% and-14.21% compared with those of Cd treatment; the concentrations of Cd treated by Cd + B, F0.1%, F0.3% and F0.6% in the grains are respectively increased by 20.12%, 32.34%, 14.73% and 8.93% compared with those of Cd treatment. In conclusion, boron promotes the absorption of cadmium by all tissues except glumes in the mature period of the wheat. Therefore, the boron fertilizer is applied before the jointing stage, preferably before seeding, in the seedling stage and/or in the jointing stage, and can inhibit the absorption, storage and transport of cadmium in winter wheat.
The application of the boron fertilizer in inhibiting the absorption, storage and transport of cadmium in winter wheat and the inhibiting method thereof are described in detail with reference to the following examples, but the application and the inhibiting method are not to be construed as limiting the scope of the invention.
Example 1
1. Plant material and growth pathway
Potting experiments were performed in the maozhuang science and education garden of the university of agriculture in Henan. Each plastic pot (80 cm, width, height, 60cm, 34cm) was filled with 100kg of air-dried and sieved topsoil. The background value is 10.31g/kg of organic matter, 65.77mg/kg of quick-acting nitrogen, 10.56mg/kg of quick-acting phosphorus and 80.42mg/kg of quick-acting potassium2.53 mS.cm for electrolyte leakage-1pH 7.96. The wheat seeds are Bainong 207. Sowing on 19 days 10 months and 19 days 2019, finally, 248 seedlings emerge from each pot, and the sowing is repeated for 4 times by adopting a completely random design.
The test is carried out by 6 treatments of CK, Cd + B, F0.1.1%, F0.3% and F0.6%. Control treatment (CK) N (217 kg/hm) was added with urea, potassium dihydrogen phosphate and potassium chloride, respectively2)、P2O5(88kg/hm2)、K2O(91kg/hm2) Adding trace elements into each pot by 100mL, wherein the nutrient solution contains 46.2mM H3BO3(the soil after addition contained 0.5mg B/kg of soil) 9.1mM MnCl2·4H2O、0.8mM ZnSO4·7H2O、0.3mM CuSO4·5H2O, 100mM FeNaEDTA and 0.2mM (NH)4)6Mo7O24·4H2O, 4mg Cd/kg Cd (NO) was added to the remaining 5 treatments3)2·4H2The soil of O, Cd + B treatment (1.5mg B/kg soil) was further added with 2 times boron on the basis of Cd treatment. The foliar spray refers to 750g boric acid per hectare in a field test, wherein 0.036g boric acid is mixed with 36mL distilled water for treating F0.1%, 0.108g boric acid is mixed with 36mL distilled water for treating F0.3%, and 0.216g boric acid is mixed with 36mL distilled water for treating F0.6%. In order to avoid the harm of boron to leaves, foliar boron fertilizers are sprayed twice in 24 days in 3 and 4 and 7 days in 2020 respectively.
2. Sample harvesting
Wheat samples were collected at seedling stage (73 days after sowing) and at jointing stage (162 days after sowing), and harvested by rooting and overground part. Collected and divided into three parts of root, stem and ear in the flowering period (176 days after sowing), and collected into four parts of root, stem, shell and seed in the mature period (210 days after sowing). The collected samples were rinsed with clean tap water and distilled water. A portion of the sample was taken back to the laboratory and dried to constant weight at 75 ℃ for elemental analysis. Another part of the samples harvested at the seedling and flowering stages were stored in liquid nitrogen and taken to the laboratory for gene expression analysis. And (5) naturally drying the sample harvested in the maturation stage, recording the grain weight, and measuring the thousand grain weight.
3. Elemental analysis
Harvesting the dried sampleGrinding and sieving, placing about 150mg sample into a digestion tube, adding HNO3/HClO4(4/1, v/v) mixed acid. The concentrations of cadmium, iron (Fe), phosphorus (P), zinc (Zn), iron (Fe), manganese (Mn), copper (Cu) and boron in the digestion solution were measured by inductively coupled plasma emission spectrophotometry (ICP-OES, Varian 710ES, USA).
4. Total RNA extraction and Gene expression analysis
Total RNA was extracted from 0.1g of fresh, rapidly frozen rhizome tissue using TRIzol reagent (Life Technologies). RNA yield was determined using a NanoDrop 2000 spectrophotometer (Thermo Scientific, USA) and integrity was assessed by ethidium bromide stained agarose gel electrophoresis. First strand cDNA was synthesized from 1. mu.g total RNA using the ReverTraace qPCR RT kit (FSQ-101, TOYOBO). Use of
480 II real-time PCR instrument (Roche, Switzerland) performed real-time PCR, 10. mu.L of PCR reaction mixture comprising 1. mu.L of cDNA, 5. mu.L of 2 XPerfect StartTM Green qPCR Supermix, 0.2. mu.L of forward primer, 0.2. mu.L of reverse primer and 3.6. mu.L of nuclease-free water. Reactions were incubated in 384 well optical plates (Roche, Switzerland) at 94 ℃ for 30s, then at 94 ℃ for 5s and at 60 ℃ for 30s, and each sample was analyzed in triplicate.
The 6 candidate gene primers of wheat are as follows:
TCONS1113-F(SEQ ID NO.7)：5`-GCTCGATTGGATGGTCAG-3`；
TCONS1113-R(SEQ ID NO.8)：5`-ATCACCAATGCAGGCTATG-3`；
TCONS5200-F(SEQ ID NO.9)：5`-ACCATCAGATCAAGCGGA-3`；
TCONS5200-R(SEQ ID NO.10)：5`-TTCAGCTGCTTCAGGACC-3`；
TRIAE1060-F(SEQ ID NO.11)：5`-CGGTCAAGTGGTTCAACG-3`；
TRIAE1060-R(SEQ ID NO.12)：5`-GGACTGGTGGACGAAGAG-3`；
TRIAE5370-F(SEQ ID NO.13)：5`-GCTCGATTGGATGGTCAG-3`；
TRIAE5370-R(SEQ ID NO.14)：5`-ATCACCAATGCAGGCTATAAC-3`；
TRIAE5770-F(SEQ ID NO.15)：5`-TGCTCAGGCTCCATTTCC-3`；
TRIAE5770-R(SEQ ID NO.16)：5`-TGACAGAGTTCTGGTTGGGTA-3`；
TRIAE5660-F(SEQ ID NO.17)：5`-GCGGCACGCACTTGAAATA-3`；
TRIAE5660-R(SEQ ID NO.18)：5-CGCCTCCTTGATCGTCTC-3`；
GAPDH-F(SEQ ID NO.19)：5`-AAGGCTGTTGGCAAGGTG-3`；
GAPDH-R (SEQ ID NO. 20): 5 '-GTGGTCGTTCAGAGCAATCC-3'. Relative transcript levels were calculated using a dual standard curve method.
5. Statistical analysis
Data were analyzed using the SPSS statistical software package (17 th edition, SPSS inc., Chicago, IL, USA), all data expressed as mean ± Standard Deviation (SD). And performing one-factor variance analysis on biomass, yield, thousand kernel weight and Cd content, and performing two-factor variance analysis on gene expression. The Least Significant Difference (LSD) test was used for significance comparisons at P <0.05 level.
6. Results
6.1 Effect of different treatments on Dry weight and yield of wheat
As shown in Table 1, the difference of the dry weight of the wheat in the seedling stage is not significant when CK, Cd and Cd + B are treated, and the dry weight of Cd and Cd + B is respectively increased by 40.56% and 49.40% compared with CK treatment. The accumulation of dry matters in the mature period has a similar trend, and the accumulation of the dry matters can be increased by spraying boron fertilizer on the leaf surfaces. Furthermore, the addition of Cd and B had no significant effect on yield. Thousand kernel weight was significantly increased by Cd + B treatment and F0.1% treatment compared to the control, by 21.99% and 22.28%, respectively. The thousand grain weight of Cd + B treatment and F0.1% treatment is respectively and obviously improved by 25.99% and 26.29% compared with Cd treatment. The result shows that boron and cadmium can promote the accumulation of dry matters of wheat and increase the yield, and the thousand grain weight can be obviously improved by applying boron.
TABLE 1 Dry winter wheat quality, yield and thousand kernel weight under different treatments
| CK | Cd | Cd+B | F0.1％ | F0.3％ | F0.6％ | |
| Dry matter weight g in seedling stage | 0.62±0.02a | 0.88±0.16a | 0.93±0.11a | |||
| Dry matter weight g in maturation period | 137.4±4.33a | 176.5±7.04a | 167.5±20.05a | 160.5±18.19a | 176.0±13.57a | 149.5±10.01a |
| Yield g/basin | 166.3±7.8a | 263.0±19.5a | 267.4±27.8a | 209.6±37.4a | 282.5±32.3a | 225.1±20.6a |
| Thousand seed weight g | 34.23±1.43bc | 33.14±2.31c | 41.75±0.61a | 41.85±0.77a | 40.11±1.17ab | 39.13±1.89abc |
Data represent mean ± standard error of quadruplicate. Different lower case letters indicate significant differences between different treatments (LSD multiple range test, P < 0.05).
6.2 Effect of different treatments on cadmium content in wheat seedling stage
And (4) measuring the cadmium content in the seedling stage, the jointing stage, the flowering stage and the mature stage. In the seedling stage, the wheat has high cadmium concentration obviously (figure 1), which shows that the wheat can absorb cadmium in the early growth stage. The Cd + B treatment (cadmium content 3.13mg/kg) was 43.90% lower (P <0.05) than the Cd treatment (cadmium content 5.58 mg/kg). The result shows that the boron application can obviously reduce the absorption of Cd in the seedling stage of wheat.
6.3 Effect of different treatments on cadmium content in wheat jointing stage
And (4) measuring the Cd content of the roots and the overground parts of the wheat in the jointing stage. As a result, the Cd content in the roots was higher than in the aerial parts, as shown in FIG. 2 and Table 2. The Cd contents of the Cd + B, F0.1.1% and F0.3% treated roots were reduced by 74.59% (P <0.05), 46.63% (P <0.05) and 72.02% (P <0.05), respectively, compared to Cd treatment. Overground Cd + B, F0.1.1%, F0.3% and F0.6% treatments were reduced by 52.11% (P <0.05), 39.37% (P <0.05), 47.46% (P <0.05) and 72.02% (P <0.05), respectively, over Cd treatment. These results indicate that the Cd content in the wheat root and the overground part during jointing is significantly reduced whether the boron soil or the boron leaf surface is applied.
TABLE 2 cadmium content (mg/kg) at different parts of the jointing stage
6.4 Effect of different treatments on cadmium content in wheat flowering time
As shown in FIG. 3 and Table 3, the cadmium content in different parts of the flowering phase is in the order of root > ear > leaf. In the root system, Cd + B, F0.3.3% and F0.6% treatments were 174.55% (P <0.05), 38.76% and 413.46% (P <0.05), respectively, higher than Cd treatment. In leaves, Cd + B, F0.1% and F0.6% treatments were 178.29% (P <0.05), 51.20% and 67.10% respectively higher than Cd treatment. The Cd contents of the grains treated by Cd + B and F with the concentration of 0.6 percent are respectively increased by 106.1 percent (P is less than 0.05) and 15.55 percent compared with the Cd treatment. The result shows that the cadmium content of different parts of the wheat can be improved by applying the boron fertilizer in the flowering phase of the wheat, and the increase range of the boron application amount of soil is larger than that of the boron spraying on leaf surfaces.
TABLE 3 Effect of different treatments on cadmium content in wheat flowering phase
6.5 Effect of different treatments on the cadmium content of wheat in the maturation phase
The Cd contents of different parts in the maturation period are in the order of root, grain, stem and shell. In the root system, Cd concentrations increased 90.38% (P <0.05), 58.72%, 49.59% and 107.5% (P <0.05) for Cd + B, F0.1.1%, F0.3% and F0.6% treatments, respectively, over Cd treatments (A in FIG. 4). In the stems, Cd concentrations of Cd + B, F0.1.1%, F0.3%, and F0.6% treatment increased by 7.81%, 25.53%, 2.04%, and 8.36%, respectively, over Cd treatment (B in fig. 4). In glumes, Cd concentrations for Cd + B, F0.1.1%, F0.3%, and F0.6% treatments were reduced by 29.77%, 28.66%, 23.95%, and-14.21%, respectively, over Cd treatment (C in fig. 4). In the grain, the concentrations of Cd treated with Cd + B, F0.1.1%, F0.3%, and F0.6% were respectively increased by 20.12%, 32.34%, 14.73%, and 8.93% over Cd treatment (fig. 4D). In conclusion, boron promotes the absorption of cadmium by all tissues except the glumes during the mature period of wheat (table 4).
TABLE 4 cadmium content at different parts of maturation period
6.6 Effect of different treatments on Cd content in soil in maturation stage
And (4) measuring the content of Cd in the soil after harvesting in the mature period of the wheat. Cd. Cd concentrations of 1.24mg/kg, 1.19mg/kg, 1.39mg/kg, 1.26mg/kg and 2.47mg/kg were treated with Cd + B, F0.1.1%, F0.3% and F0.6% (FIG. 5). The Cd concentrations were 11.72%, 1.12% and 98.95% higher than Cd treatment at F0.1%, F0.3% and F0.6%, respectively (P < 0.05). The result shows that the spraying of the boron fertilizer on the wheat leaf surfaces can reduce the absorption of cadmium, and the treatment effect is best at high concentration.
6.7 Effect of different treatments on relative expression levels of 6 Cd transporter genes in seedling stage
Real-time quantitative PCR analysis showed that these 6 genes were predominantly expressed in roots during the seedling stage (fig. 6 and table 5). Among these genes, Cd + B treated TCONS1113, TRIAE5370, TRIAE5770 and TRIAE5660 showed reduced relative expression levels in roots and aerial parts compared to Cd treatment, while TCONS5200 and TRIAE1060 were increased. The relative expression levels of TCONS1113 in Cd + B treated roots and leaves were significantly reduced by 24.15% and 43.47%, respectively, compared to Cd treatment (a in fig. 6).
The relative expression levels of Cd + B treated triee 5370 in roots and leaves were significantly reduced by 36.09% and 40.67%, respectively, compared to Cd treatment (D in fig. 6).
The relative expression levels of Cd + B treated triee 5770 in roots and leaves were reduced by 76.48% (P <0.01) and 20.54% (E in fig. 6), respectively, compared to Cd treatment.
The relative expression levels of Cd + B treated triee 5660 in roots and leaves were significantly reduced by 37.22% and 89.72%, respectively, compared to Cd treatment (F in fig. 6). Correlation analysis found (table 6) that cadmium concentration was negatively correlated with the expression of TCONS1113, triee 1060 and triee 5370 in roots and aerial parts. In addition, cadmium concentration is negatively correlated with the expression of TCONS5200 and TRIAE5770 in the aerial parts. Expression of these genes may play an important role in regulating Cd uptake.
TABLE 5 relative expression of genes at seedling stage
TABLE 6 correlation coefficient of Cd concentration and Cd transporter gene expression in seedling stage
"R" represents a root and "S" represents an aerial part
6.8 Effect of different treatments on the relative expression levels of 6 Cd transporter genes in flowering phase
At anthesis, these genes had different tissue-specific expression and differential response to boron (fig. 7 and table 7). These 6 genes were also expressed mainly in roots, whereas TRIAE5770 and TRIAE5660 were hardly detectable in leaves and ears at wheat flowering. Of these 6 genes, Cd + B treated TRIAE1060 and TRIAE5770 tended to increase relative expression levels in the root system compared to Cd treatment, while TCONS1113, TCONS5200, TRIAE5370 and TRIAE5660 decreased. Furthermore, 6 genes (except TRIAE 5770) all increased with increasing boron concentration sprayed, especially in roots and leaves. Meanwhile, cadmium concentration was negatively correlated with the expression of TCONS5200(P <0.01), TCONS1113, TRIAE5370 and TRIAE1060 in roots (table 8), with the expression of TRIAE5660, TRIAE5770 and TCONS1113 in leaves, and with the expression of TRIAE5370 and TRIAE1113 in ears. The cadmium concentration is significantly and positively correlated with TRIAE5770(P <0.01), TRIAE5660(P <0.01) and TRIAE1060(P < 0.05). Expression of these genes may play an important role in regulating Cd uptake and transport.
TABLE 7 Gene expression level at flowering time
TABLE 8 correlation coefficient of Cd concentration and Cd transporter gene expression in flowering phase
And indicate significance of difference at 0.01< P <0.05 and P <0.01 levels, respectively
6.9 Effect of different treatments on the content of various nutrient elements in wheat grains
The results are shown in fig. 8 and table 9, with Cd treatment reducing the Fe, Mn, Cu and B content in the grain compared to the control. In addition, the 6 elements (phosphorus, zinc, iron, manganese, copper and boron) content of the Cd + B treatment were all lower than that of the Cd treatment. The content of P and Mn in the grains is reduced by spraying the fertilizer B, wherein the content of P is obviously reduced by 13.61%, 18.18% and 19.21% (A in figure 8), and the content of Mn is respectively reduced by 35.97%, 47.55% (P is less than 0.05) and 35.73% (D in figure 8) in treatment when F is 0.1%, F is 0.3% and F is 0.6% compared with Cd treatment. Elemental correlation analysis showed that Cd is negatively correlated with Cu, Fe, Mn (P <0.05), P (Table 10). Cu, Mn (P <0.05), P (P <0.05) and Zn (P <0.05) in the grains are in extremely obvious positive correlation. Mn and Fe are obviously in positive correlation (P <0.05) and P (P < 0.05). B has no significant correlation with other elements.
TABLE 9 mineral content of seeds in maturation stage
TABLE 10 correlation coefficients between different elements in grain
And indicate differential significance at levels of 0.01< P <0.05 and P <0.01, respectively.
The foregoing is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and decorations can be made without departing from the principle of the present invention, and these modifications and decorations should also be regarded as the protection scope of the present invention.
Sequence listing
<110> Henan university of agriculture
Application of <120> boron fertilizer in inhibiting absorption, storage and cadmium transfer of winter wheat and inhibition method
<160> 20
<170> SIPOSequenceListing 1.0
<210> 1
<211> 773
<212> DNA
<213> Triticum aestivum
<400> 1
tcaaatcacg accaacacgt accggatctt gaccgaccga accattcagt gctcgcgctc 60
actcacgcat catagccaag ttaagcggga aggaaggaag gaaggaagcc atgtctgccg 120
cggagggagc cgtcgtgttc agcgaggaga aggaggcgct ggtgctcaag tcatgggcca 180
tcatgaagaa ggattccgcc aaccttgggc tccgcttctt cctcaagatc ttcgagatcg 240
cgccgtcggc gaggcagatg ttcccgttcc tgcgcgactc cgacgtgccg ctggagacca 300
accccaagct caagacccac gccgtgtccg tcttcgtcat gacgtgcgag gctgctgcgc 360
agctgcggaa agccgggaag atcaccgtca gggagaccac cctgaagagg ctgggcggca 420
cgcacttgaa atacggcgtg gcagatggcc actttgaggt gacgcggttc gctctgctcg 480
agacgatcaa ggaggcgctt ccggcggaca tgtgggggcc ggagatgagg aacgcgtggg 540
gcgaggccta cgaccaactg gtcgcggcca tcaagcaaga gatgaagccc tctgagtagc 600
tcatccattg tactcatatc atatgccacg caacttccgt ccatatccgt ccaactttcg 660
ttgcttgacc ggttcactca tgtcaccata ttgtgtttgt attgtgtgtt tacgtgtact 720
aacgcatatt gtaaaatggg cattcaataa aggaacaaat tgtgcatgag att 773
<210> 2
<211> 996
<212> DNA
<213> Triticum aestivum
<400> 2
atggagtact ctcaccgcag caggttgctc ctggtgtgct cagttcttgt gctgtgcctc 60
gtaacccgag gcgcgaggtg cgatgagtta accagcgatt tctacgactg gacatgcccc 120
ggtgtgtatg atgtcgtcca gcagcaagtg ttttctgcca tgagggaaga gcccaggatg 180
ggagcctccc tgctcaggct ccatttccat gactgctttg tcaatgggtg tgatggttca 240
atcttgttgg atggcaccga cggcgagaaa tttgcactac ccaaccagaa ctctgtcaga 300
gggtacgagg tcatcgatgc aatcaaggcc gacctcgaaa acatgtgccc tggggtggtg 360
tcctgtgctg acattgtggc cattgcagct ggctatggag tactctttag tggagggcct 420
tactatgatg ttcttctggg gagaagggac ggtctgaagg cgaatcagac aggagctgac 480
aacggcctgc cctcgccgtt cgaaccgatc agctccatcg tacagaagtt tgccgatgtc 540
ggcctcgaca cgaaagacgt tgtggtcctg tcaggtgccc acacgatcgg gcgagcccgg 600
tgcgtgctgt tcagcgaccg gctgacgtcc accaagagct cggccgaccc gacgctggac 660
gccaccatgg ccgccgacct gcagaagctc tgcaccggcg gcgacggcaa ccagaccacc 720
gcgctggacg tgagctccgc ggatgtgttc gacaagcagt actaccacaa cctgcttagc 780
aagaagggcc tcctgacctc tgaccagggc ctcttctccg ccgacgagga cgtcgtcgcc 840
agcaccacaa aggcgttggt gcagacgtac agcgacgacg gcgagcagtt cttctctgac 900
ttcggcgcgt ccatggtgaa gatggggagc ataccgttgc cagcgggatc cgccggcgag 960
atccgctgca actgcagggt tcccaataaa aaatga 996
<210> 3
<211> 2387
<212> DNA
<213> Triticum aestivum
<400> 3
ctgcaagcca agccaagccc aggtataaat ggaggacgtc ttcccctccc agaagaccac 60
caaaccagca cacgtcaacg gaacagctta gcaaaatccc caccaaggtg tagaatattc 120
cccaaaaccc cacgtcccct ccctctgccg ccgctcccac tccaccagtg tgccacccgg 180
atcgatcgat cgatcgacag cgatggacac ccacatcggc tccgtcgacg ggccgtcgcc 240
ggcggcggtg aacggcgcgg tcggctgccc ggcgtccgcg ccggggtgcc cgatcatgtc 300
ctcccacccc gtggtctccg caggcgaggc gtcgctgggg cgccacctgg cgcgccgcct 360
cgtgcaggtc ggcgtcagcg acgtcttcgc cgtgcccggg gacttcaacc tcacgctgct 420
cgaccacctc gtcgacgagc ccgggctgcg cctcgtcggc tgctgcaacg agctcaacgc 480
tggctacgcg gccgacggct acgcgcgggc ccgcggcgtc ggcgcctgcg cggtcacctt 540
caccgtcggc ggcctcagcg tgctcaacgc catcgccggc gcctacagcg agaacctgcc 600
cgtcatctgc atcgccggcg ggcccaactc caacgactac ggcaccaacc gcatcctcca 660
ccacaccatc ggcatcccgg acttctcgca ggagctgcgc tgcttccaga ccgtcacctg 720
ccaccaggcg gtggtgacca acctggacga cgcgcacgag cagatcgaca cggccatcgc 780
cacggcgctc agggagagca agccggtgta cctcagcatc agctgcaacc tccccgggct 840
acctcacccc accttcaccc gtgacccagt ccctttcttc ctcgccccca ggatgagcaa 900
caagatgggg ctcgaggctg cagtggaggc aaccgtcgag ttcctgaaca aggcggtgaa 960
gccagtgctt gtcgccggcc ccaaactgcg cgtggccaag gcggggaagg ccttcgtcga 1020
ccttgtggac gccagtggct atgcctatgc tataatgcca tcggccaagg gctttgtgcc 1080
agagacgcac ccccacttcc tcggcaccta ctggggcgcc gtcagcacgg ccttctgcgc 1140
cgagatcgtc gagtcggccg acgcctacct cttcgcaggc cccatcttca acgactacag 1200
ctctgttggc tactcgttcc tgctcaagaa ggacaaggcc atcatcgtgc agcctgagcg 1260
tgtcatcgtc gggaacggcc cggcattcgg ctgcgtcatg atgaaggagt acctgtctgc 1320
attggccaag cgggttcaga agaacaccac cgcctacgag aactacaaga ggatcttcgt 1380
gcctgagggc catccgctga agggcgaggc gaacgagccg ctgcgtgtca atgtgctctt 1440
caagcacatc caggacatgc tgacgggtga cagtgcagtg ctcgctgaga ccggtgactc 1500
ctggttcaac tgccagaagc tcaagctgcc cgagggctgc gggtatgaat tccaaatgca 1560
gtatggctcg attggatggt cagtgggtgc attgctcggg tacgcgcaag gggcgagtga 1620
caagcgtgtc atagcctgca ttggtgatgg gagcttccag gtgacagcac aggatgtgtc 1680
aactatgctg cggtgcgaac agaacagcat aatcttcctg atcaacaatg gcgggtacac 1740
gatcgaggtg gagatccacg acgggcctta caatgtcatc aagaactgga actacaccgc 1800
ccttgtggag gccatccaca acggggaggg caaatgctgg actgccaagg tgaagtgcga 1860
ggaggagctg acggcggcga tagagacggc gctgggggag aagaaggact ctctgtgctt 1920
catcgaggtg atcgcgcaca aggacgacac cagcaaagag cttctggaat ggggctccag 1980
ggtctctgct gccaactcca ggccacccaa cccccagtag accaagcacg cctgcatcac 2040
tagtagtcgc agccagcagc acgttgccat caaataatgt tgaacagttt atctttgctc 2100
ttctgttatg ttcttagttt tgtgcctact agttactgta cttgtgagct gtggccatcg 2160
cctgtgttct atcctgccaa tgctaagaag atcatgagat atttttagct tgccccagct 2220
atttgtgtgt gtgtgccttt ttctttcaaa atgtgcttgc caatgtagtt gaaaatcacg 2280
agctactcct gagtggctcc ttgaggaatc tagaagcgca atgctacatg catgaacaca 2340
gaatcttgca gataggatac ttttggtaag aaattgagct gatagtg 2387
<210> 4
<211> 2402
<212> DNA
<213> Triticum aestivum
<400> 4
agaccaccaa accagcacac gtcaacggaa cagcttagca aaatccccac caaggcctag 60
aatattcccc aaaaccccac gtcccctctg ccgccgctcc ccactccacc agtgtgccac 120
ccggatcgat cgatcgatcg acagcgatgg acacccacgt cggctccgtc gacgggccgt 180
ccccggcggc ggtgaacggc gcggtgggct gcccggcgtc cgcgccgggg tgcccgatca 240
tgtcctccca cccggtggtc tccgcaggcg aggcgtcgct ggggcgccac ctggcgcgcc 300
gcctcgtgca ggtcggcgtc agcgacgtct tcgccgtgcc cggggacttc aacctcacgc 360
tactcgacca cctcgtcgac gagcccgggc tgcgcctcat cggctgctgc aacgagctca 420
acgcgggcta cgcggccgac ggctacgcgc gggcccgcgg cgtcggcgcc tgcgcggtca 480
ccttcaccgt cggcggcctc agcgtgctca acgccatcgc cggcgcctac agcgagaacc 540
tgcccgtcat ctgcatcgcc ggcgggccca actccaacga ctacggcacc aaccgcatcc 600
tccaccacac catcggcatc ccggnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 660
nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn nnnnnnnnnn 720
nnnnccacgc acgcgcgcgg tggtgaccaa cctggacgac gcgcacgagc agatcgacac 780
ggccatcgcg acggcgctca gggagagcaa gccggtgtac ctcagcatca gctgcaacct 840
ccccgggcta cctcacccca cctttacccg tgacccagtc cccttcttcc tcgcccccag 900
gatgagcaac aagatggggc tcgaggctgc agtggaggca actgtcgagt tcctgaacaa 960
ggcggtgaag ccagtgcttg tcgccggccc caaattgcgc gtggcgaagg cggggaaggc 1020
cttcgtcgac ctcgtggacg ccagtgggta tgcctatgct ataatgccat cggccaaggg 1080
ctttgtgcca gagacgcacc cccacttcct cggcacctac tggggcgccg tcagcaccgc 1140
cttctgcgcc gagattgtgg agtcggccga cgcctacctc ttcgcaggcc ctatcttcaa 1200
cgactacagc tctgttggct actcgttcct gctcaagaag gacaaggcca tcatcgtgca 1260
gcctgagcgt gtcatcgtcg ggaacggccc ggcattcggc tgcgtcatga tgaaggagta 1320
cctgtctgca ttggccaagc gggttcagaa gaacaacact gcctacgaga actacaagag 1380
gatcttcgtg cctgagggcc atccgctgaa gggcgaggcg aacgagccgc tgcgtgtcaa 1440
tgtgctcttc aagcacatcc aggacatgct tacgggtgac agtgcagtgc tcgccgagac 1500
cggtgactcc tggttcaact gccagaagct caagctgccc gagggctgcg ggtatgaatt 1560
ccaaatgcag tatggctcga ttggatggtc agtgggtgca ttgctcgggt acgcgcaagg 1620
tgcaagcgac aagcgtgtta tagcctgcat tggtgatggg agcttccagg tgacagcaca 1680
ggatgtgtca actatgctgc ggtgcgaaca gaacagcata atcttcctga tcaacaatgg 1740
cgggtacacc atcgaggtgg agatccacga cgggccttac aatgtcatca agaactggaa 1800
ctacaccgcc cttgtggagg ccatccacaa cggggagggc aaatgctgga ctgccaaggt 1860
gaagtgcgag gaggagctga cggcggcgat agagacggcg ctgggggaga agaaggactc 1920
tctgtgcttc atcgaggtga tcgcgcacaa ggacgacacc agcaaagagc ttctcgagtg 1980
gggttccagg gtctctgctg ccaactccag gccacccaac ccccagtaga ccaagcaaac 2040
ttgcctgcat cagtagtagt agcagcagca gcacgcacgt tgcccatcaa ataatgtgaa 2100
cagtttatct ttgctcttct gttatgttct tagttttgtg cctactagtg acttgtgagc 2160
tgtggccatc gcctgtgttc tatcatgcca atgctacgaa gattatgaga tatttctagc 2220
ttgcctcggc tatttgtgtg tgtgccttct tctttcaaaa tgtgcggcca atatttgaaa 2280
atcacgagct actcctgagt ggctcctcga ggaatctaga agcgcaatcc aacatgcatg 2340
gactcggaat ctagaagcgc aatccaacat gcatggactc ggaatagaat cttgcaaata 2400
tg 2402
<210> 5
<211> 774
<212> DNA
<213> Triticum aestivum
<400> 5
atggcgatgg aggcgcggat gaaggggacg gtcaagtggt tcaacgacac caaggggttc 60
ggcttcatct cccccgacga cggcagcgag gacctcttcg tccaccagtc ctccatcaag 120
gccgacggct tccgctcgct ggccgagggc gaggtcgtcg agttcgccgt ctcggagggc 180
gacgacggcc gcaccaaggc cgtcgacgtc accggccccg acggatcctt cgtgcagggc 240
ggcgcgggcg gtggcggtgg cgggggcggc ttcggatccc gtggcggtgg cggatctcgc 300
ggcggcgggg gcttcggcgg ccgcggcggg gacggatctg gcggctacgg aggtgggtac 360
ggtggtggcg gtggcggagg ctggggcggc cagaggagat ccggcggtgg aggcgccggt 420
ggggcctgct tcaagtgcgg ggagcctggc cacatggcca gggactgctc ggtcaacggc 480
cccgctggtg gcggcggcgg cggcggctgc tacaagtgcg gcgagcaggg ccacatcgcc 540
agggactgct tcaacggcgg cggcggcgga ggaggcggcg gctacggcgg cggcggcggc 600
ggcaactgct acaactgcgg cgagccgggc cacatcgcga gggactgccc caccagcagc 660
ggctttggcg ggggcggcgg cgggaggttc ggcggaggag gcggcggcgg cggcgaccgc 720
tcctgctaca actgcggcga gcccggccac atctcccgcg actgcaccaa gtga 774
<210> 6
<211> 1194
<212> DNA
<213> Triticum aestivum
<400> 6
atgcaaagac gatccaaacc cctgtctctt tattgctaca ccaacattgc acacagaaaa 60
aagcagcatg ccaagaaacc caaatacgat tggctacggc ctacggccat gccatgcatg 120
cacgctaaca caacgcacgg gcgggcagct agcctgcttg ccgaccaccg aaccatcaga 180
tcaagcggaa gcggcgaagc cctcctcctc gacggcggcg tccacggcag cgtcggtgcc 240
ctgggcgttg gtcctgaagc agctgaagcg ccggatctcg cccacgttgc cggccggctt 300
gggcaccctg gcgagcttct ccatggacct ggcgaactgc ccgaagaagg cctccttgct 360
ctcggcgaac cgcttcacga tgggcgcggt ggtcttgttc ttcacgaggg ccatgtcgga 420
ggtgaacacg ccctggttga aacccagcgc cttgtagtac gcgttgtcga acaggtccgg 480
ggtgatcacg tccaggttct gcagccggtc cgggtgcttg ctgcagttgg cggcgagctt 540
gcgggagaag gtgtcgtcgg cgcgctggga ccggtcttcg aagctgccgc actgcgaccg 600
cccgatggtg tgcgcgccgg agagcgccac gaggtcggcc acgtcgccga ggcccctggt 660
gccgaaggac tccaccagcg tcgccacgct ggccgtgttg ggcgccggga ggtcgaacac 720
cttggcctgc gccgcggggg cgaggctgtc gaggtttccc tgcggcacgt tgaaccaggg 780
cccgccggag atgacgatga tgtcgcgggt ggcgagcagg gtgatgtcgg cgcaggagac 840
gacgggcccg cacacggcgt gcgccctggc gcggatggac tcgatgagct gcatggcgcg 900
cggctggatg gtgaggttgg ggccgagcgc ggtctcgcgg gcggccgtgt tgttgaggag 960
gatggaggcg tcgcagccct gcgggaagca gtcgtggaag tagaggcgga gcatgcccgc 1020
cgccacggcc acctcctgcc gcagcgccgc ctccacggac gaccacacga tgccgtccag 1080
ctgcgtgcac gacgccgcgt ggaagtccgg ggaggcgtcg ccggtgaaga cgggcaggga 1140
catggccggg gagagcagcg cggccgcggt gaccaggacg gccagtattg ctga 1194
<210> 7
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 7
<210> 8
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 8
<210> 9
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 9
<210> 10
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 10
<210> 11
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 11
<210> 12
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 12
<210> 13
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 13
<210> 14
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 14
atcaccaatg caggctataa c 21
<210> 15
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 15
<210> 16
<211> 21
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 16
tgacagagtt ctggttgggt a 21
<210> 17
<211> 19
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 17
<210> 18
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 18
cgcctccttg atcgtctc 18
<210> 19
<211> 18
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 19
<210> 20
<211> 20
<212> DNA
<213> Artificial sequence (artificial sequence)
<400> 20
gtggtcgttc agagcaatcc 20
Claims (6)
1. The boron fertilizer is applied to inhibiting absorption, storage and cadmium transfer of winter wheat.
2. The use of claim 1, wherein the boron fertilizer inhibits cadmium absorption, storage and transport in winter wheat during seedling stage by regulating expression of genes related to cadmium absorption, storage and transport in winter wheat.
3. The use of claim 2, wherein the genes involved in cadmium uptake comprise TRIAE5660, TRIAE5770 and TCONS 5200; the nucleotide sequence of the TRIAE5660 is shown as SEQ ID NO.1, and the nucleotide sequence of the TRIAE5770 is shown as SEQ ID NO. 2; the nucleotide sequence of the TCONS5200 is shown as SEQ ID NO. 6;
the related genes for storing cadmium comprise TCONS1113 and TRIAE 5370; the nucleotide sequence of the TCONS1113 is shown as SEQ ID NO. 3; the nucleotide sequence of the TRIAE5370 is shown as SEQ ID NO. 4;
genes associated with cadmium transport include triee 5770 and triee 1060; the nucleotide sequence of the TRIAE1060 is shown as SEQ ID NO. 5.
4. A method for inhibiting absorption, storage and transport of cadmium in winter wheat is characterized by comprising the following steps: before sowing winter wheat, applying boron fertilizer in the form of base fertilizer, or before jointing stage, spraying boron fertilizer on leaf surface.
5. The method as claimed in claim 4, wherein the cadmium content of the winter wheat is 0-4 mg/kg.
6. The method of claim 4, wherein the boron fertilizer is H3BO3The total amount of the fertilizer applied to each mu is 1.0-1.5 kg.
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