CN112390868A - Mulberry calmodulin protein MmCML37 and application thereof - Google Patents
Mulberry calmodulin protein MmCML37 and application thereof Download PDFInfo
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Abstract
The invention discloses a mulberry calmodulin protein MmCML37 and application thereof. The gene sequence and the amino acid sequence of the mulberry calmodulin protein MmCML37 are respectively shown as SEQ ID NO.1 and SEQ ID NO. 2. According to the invention, through the research of the calmodulin-like protein MmCML37 of the mulberry under the calcium stress, the mulberry genome is perfected, and the expression analysis and the application of the calmodulin-like protein of the mulberry under the calcium stress are provided. The mulberry calmodulin protein MmCML37 gene is transferred into agrobacterium tumefaciens GV3101 by using VIGS technology, the expression of the gene is reduced by using the virus silencing principle, and then the physiological and biochemical indexes of transgenic plants are detected to research the effect of the mulberry calmodulin protein MmCML37 on improving the stress resistance of plants, thereby providing an important molecular basis for researching a mulberry calcium stress resistance regulation mechanism and cultivating new varieties.
Description
Technical Field
The invention belongs to the field of plant genetic engineering, and particularly relates to a mulberry calmodulin protein MmCML37 and application thereof.
Background
Calcium is a major element necessary for plant growth and development, is an intracellular and extracellular messenger molecule capable of transducing various physiological processes, and has effects at many sites. Calcium is not only in central position in plant growth and development, photosynthesis and related enzyme regulation, but also is an important influence factor of plants to environmental stress. In China, about 1/3 land is karst landform, the content of calcium ions in soil in the regions is more than 3 times that of acid soil, and high-concentration calcium ions can inhibit seed germination, influence plant photosynthesis and reduce plant growth characteristics, so that the growth of plants is inhibited, and even the distribution of plant communities is limited. However, plants in calcareous soil with high calcium content in north China can have physiological calcium deficiency, and particularly, the phenomenon of calcium deficiency of plants with high calcium demand is common. Some crops need high calcium amount, even the calcium demand is large in the whole growth and growth process, and the crops are easy to lack calcium. In order to adapt to the problems of high calcium or calcium deficiency caused by various soil environments, plants must strictly control the transport of calcium ions on the whole and cellular level. The plant species can be improved in a molecular biology way by researching the calcium ion absorption mechanism and physiological and biochemical indexes of the plant, and the calcium stress resistance of the plant can be improved.
Under the stress of calcium, physiological and biochemical indexes such as malondialdehyde, soluble protein, peroxidase activity, superoxide dismutase activity, proline content and the like in plants generate a series of changes to respond to the stress of calcium. Calcium receptors in plants are roughly classified into 3 types: calmodulin-like B subunit proteins (CBL), Calcium Dependent Protein Kinases (CDPK), calmodulin proteins (CaM) and calmodulin-like proteins (CMLs). Among them, calmodulin-like proteins (CMLs) are used as plant-specific calcium receptors, participate in various physiological processes of plants, and have important roles in plant stress tolerance.
Research shows that AtCML8 in Arabidopsis can be induced by Pseudomonas syringae and plays a key role in immunity; the resistance of the AtCML9 function-deletion mutant to salt damage and drought is improved; the AtCML20 is lost, the pore diameter of the plant stomata is reduced, and the moisture of the leaves is easier to store compared with the wild type; whereas AtCML19 plays a role in the early stages of DNA repair. GhCML11 in dry land cotton can interact with GhMYB108 to form a positive feedback mechanism, and the transcription level of GhCML11 is improved, so that the invasion of verticillium wilt is resisted. LjCML6, LjCML7 and LjCML12 in Lotus corniculatus are highly expressed in nodules, which are presumed to be involved in nitrogen fixation symbiosis.
Virus-induced gene silencing (VIGS) is a post-transcriptional gene silencing phenomenon discovered in recent years and is a natural mechanism for plants to resist virus infection. VIGS has been developed as a new reverse genetics technology for rapidly identifying plant gene functions, and a VIGS system using RNA viruses, DNA viruses, satellite viruses and DNA satellite molecules as vectors has been established so far. The VIGS does not need to construct a transgenic plant, has the advantages of short period, simple and convenient operation, quick phenotype acquisition, low cost and the like, and at present, the novel reverse genetics technology is widely applied to the research on the functions of related genes such as plant disease resistance, stress, cell signal transduction, growth and development and the like, and plays an important role in the research on plant functional genomics.
Mulberry is an important ecological economic forest, China is the earliest country in the world for developing the silkworm industry, and the comprehensive utilization of mulberry resources and the development of the mulberry industry are more and more emphasized. Fully recognizes and discovers the ecological function of the mulberry, correctly grasps the relationship between the mulberry ecological industry and the traditional silk industry, establishes various mulberry ecological industry modes and corresponding silk industry production and operation modes, develops the scientific research of the mulberry adapting to the ecological industry, and is very important for promoting the development of the mulberry ecological industry and the silk industry.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides a mulberry calmodulin protein MmCML37 and application thereof, wherein the application refers to metabolic detection and expression regulation under the condition of calcium stress, and definitely shows the feasibility of the mulberry calmodulin protein MmCML37 as a molecular marker.
In order to solve the technical problems, the invention adopts the technical scheme that:
a mulberry calmodulin protein MmCML37, wherein the amino acid sequence of the mulberry calmodulin protein MmCML37 is as shown in SEQ ID No.: 2, respectively.
The nucleotide sequence of the encoding mulberry calmodulin protein MmCML37 of claim 1 is shown in SEQ ID NO. 1.
A vector comprising the nucleotide.
A genetically engineered bacterium comprising the vector or having the nucleotide integrated into its genome.
Use of a mulberry calmodulin protein MmCML37 or a coding gene thereof as a molecular marker for identifying the calcium stress resistance of mulberry.
A method for improving plant calcium stress resistance comprises allowing mulberry to express the above mulberry calmodulin protein MmCML 37.
A molecular marker for identifying the calcium stress resistance of mulberry, wherein the molecular marker is a primer pair which specifically recognizes a coding gene of the polypeptide of claim 1; the primer pair is as follows:
MmCML37-F:ATGGGCAAGCACGTAGAGT；
MmCML37-R:TCACTGCATCATAATCCTGAAC。
the application of the molecular marker in identifying the calcium stress resistance of the mulberry.
Has the advantages that:
compared with the prior art, the mulberry calmodulin protein MmCML37 and the application thereof have the following advantages:
the mulberry calmodulin protein MmCML37 improves mulberry genome and provides expression analysis and application of mulberry calmodulin protein under different calcium content treatment conditions. Can be used for expression regulation of calmodulin-like protein in mulberry under different calcium content treatment conditions, and can improve the research of plants on calcium stress.
Drawings
FIG. 1 is a PCR map of the gene clone of Morus alba calmodulin protein MmCML37, wherein M: 500bp DNA molecular marker;
FIG. 2 shows that the MmCML37 gene is detected in different CaCl concentrations by quantitative RT-PCR2Relative expression quantity of tender leaves of treated mulberry seedlings, 440mg/L CaCl2Is a control group, and the other three groups are experimental groups;
FIG. 3 is a graph showing the results of detecting the Malondialdehyde (MDA) content of transgenic mulberry seedlings under calcium stress;
FIG. 4 is a graph showing the results of detecting proline (Pro) content of transgenic mulberry seedlings under calcium stress;
FIG. 5 is a graph showing the results of detecting superoxide dismutase (SOD) activity of transgenic mulberry seedlings under calcium stress;
FIG. 6 is a graph showing the result of detecting Peroxidase (POD) activity of transgenic mulberry seedlings under calcium stress;
FIG. 7 is a graph of the results of the detection of soluble protein (Cpr) of transgenic mulberry seedlings under calcium stress.
Detailed Description
Example 1 cloning of Mulberry calmodulin protein MmCML37 Gene
The mulberry variety to be tested is cultivated with Morus alba (Morus multicaulis) 71-1, is preserved in the mulberry garden of Zhenjiang, national germplasm of the institute of silkworm, national academy of agricultural sciences, utilizes the obtained CDs sequence of the cDNA fragment of the CML37 gene of the mulberry to design a full-length primer, and verifies that the primer sequence is as follows:
MmCML37-F:5'-ATGGGCAAGCACGTAGAGT-3'；
MmCML37-R:5'-TCACTGCATCATAATCCTGAAC-3'；
designing a primer for carrying out fluorescence quantitative analysis according to the obtained full-length sequence of the mulberry calmodulin protein MmCMML 37 gene, wherein the primer sequence is as follows:
qMmCML37-F:5'-CAAGAGCTTGAAGAGGATGCTCAGT-3'；
qMmCML37-R:5'-GCATCATAATCCTGAACTCCTCCTC-3'；
designing an upstream primer and a downstream primer of an internal reference gene according to a beta-actin gene (GeneBank accession number: DQ785808) sequence which is searched on NCBI and stably expressed in mulberry:
the upstream primer sequence beta-actin-F: 5'-AGCAACTGGGATGACATGGAGA-3', respectively;
the downstream primer sequence beta-actin-R: 5'-CGACCACTGGCGTAAAGGGA-3' are provided.
Example 2 amplification of the full-Length sequence of the Mulberry calmodulin protein MmCML37 Gene
Taking young leaves at the top of the mulberry seedlings, and extracting the total RNA of the mulberry by using an RNA kit. 1mL of RNAioso extract was prepared and chilled in a sterile centrifuge tube. Placing young leaves of mulberry seedling in a mortar, adding liquid nitrogen, grinding into powder, adding the powder into a pre-cooled extraction liquid sterile centrifuge tube, oscillating, cooling in ice for 30min, cooling at 4 ℃, rotating at 12000rpm, and centrifuging for 5 min; adding 800 μ L of the supernatant into a new sterile centrifuge tube, adding 200 μ L of chloroform, shaking sufficiently, standing on ice for 10min, centrifuging at 4 deg.C and 12000rpm for 20 min; taking 300 mu L of the supernatant into a new sterile centrifugal tube, adding equal volume of precooled isopropanol, uniformly mixing, standing on ice for 10min, 12000rpm, 4 ℃, and centrifuging for 20 min; discarding the supernatant, slowly adding 1mL of precooled 75% glacial ethanol, centrifuging at 12000rpm at 4 ℃ for 5 min; removing supernatant, drying at room temperature for precipitation for 2-5min, adding appropriate amount of ddH2O, flicking the tube wall to dissolve the RNA sufficiently.
RNA cannot be used as a template for PCR reactions and must be reverse transcribed into cDNA. Oligo (dT) using 9. mu.L of total RNA extracted as a template184 mu L of the primer is used for reverse transcription; keeping the temperature of the water bath kettle at 70 ℃ for 10min, quickly placing the water bath kettle on ice to cool for more than 2min, and centrifuging for several seconds; the 13. mu.L template RNA/oligo (dT)184 mu L of 5 XM-MLV-Buffer, 1 mu L of 10mM dNTP mixture, 1 mu L of 40U/. mu.L RNase Inhibitor, 1 mu L of 200U/. mu.L RNase M-MLV (RNaseH-), and 20 mu L of total volume are added into the denaturation solution; after centrifugation, the cDNA was obtained by cooling on ice at 42 ℃ for 1h and 70 ℃ for 15min, and stored at-20 ℃ for further use.
RT-PCR amplification was performed using the synthesized cDNA as a template and the designed MmCML37-F and MmCML37-R as primers. The PCR amplification procedure was: pre-denaturation at 95 ℃ of 3min; circulating for 35 times at 95 ℃ for 15s, 51 ℃ for 20s and 72 ℃ for 30 s; extending for 5min at 72 ℃; storing at 4 ℃.5 μ L of RT-PCR product was subjected to agarose gel electrophoresis to detect the size of the target fragment and verify whether the size of the target fragment matches the predicted size (FIG. 1). After the conditions are consistent, 50 μ L of RT-PCR product is subjected to electrophoresis by using 1% agarose gel prepared from TAE buffer solution, and gel recovery and purification of the target fragment are carried out by adopting SanPrep column type DNA gel recovery kit of Shanghai biological engineering Co. The gel recovered product was coupled to pMD by DNA ligaseTM18-T vector ligation. Coli TOP10 strain competent cells were transformed into the host strain E.coli and plated on LB solid medium plates containing 100. mu. LX-Gal, 10. mu. LIPTG and Amp. Picking out white spot culture of single colony, then sending the sample, finishing sequencing by a biological engineering (Shanghai) company Limited, performing blast analysis on a sequencing result, and comparing and verifying the sequencing result with an original sequence to obtain a gene cDNA sequence. The sequence of the mulberry calmodulin protein MmCMML 37 gene is shown in SEQ ID No. 1.
ATG*GGCAAGCACGTAGAGTACTTAGAACATGTTTTCCACTACTTTGACGAAGATGGTGATGGAAAGATATCACCCTCGGAGCTGAGGAATCGGCTAGGGTTGATGGGTGGAGAGGAGCTTATAGAGCTTAAGGAGGCGGAGGAAGCGGTGGAGTCGTTGGATTCGGACGGCGACGGGCTCTTGGGATTGGAGGATCTTGCTCGTTTGATGGAGTTAGGAGGAGGAGAGGAGGAGAAAATGAAGGATTTGAGAATGGCTTTTGAGATGTATGATGTGGAGGGAAAAGGGTTCATAACGCCCAAGAGCTTGAAGAGGATGCTCAGTAGATTGGGCGAATCGAGATCCGTTGATGAGTGCAAAGTCATGATCAACCATTTTGATTTAAATGGAGATGGTGTACTTTGCTTTGAGGAGTTCAGGATTATGATGCAGTGA*
ATG is the initiation codon; TGA is stop codon
Example 3 bioinformatic analysis of Morus calmodulin protein MmCML37 Gene
Predicting a nuclear localization sequence of a target gene and analyzing a related conserved gene sequence by using an online tool PSORT according to reported literature; carrying out homology search and comparison on the obtained product by using a Blast tool of NCBI (national center of Biotechnology information) and downloading a part of homologous amino acid sequence; carrying out homologous alignment on an amino acid sequence coded by the gene and an amino acid sequence of a protein related to a mulberry calmodulin protein MmCML37 gene by utilizing Clustal X software; the mulberry calmodulin protein MmCML37 gene was more comprehensively analyzed using DNAStar software and on-line analysis (http:// www.expasy.org), such as: prediction of amino acid and ORF encoded by the gene, prediction of isoelectric point and molecular mass of protein encoded by the gene, and the like; then, the evolutionary tree was constructed using MEGA4.1 software and the relationships of the related proteins were studied.
The amino acid sequence of the mulberry calmodulin protein MmCMML 37 gene is shown in SEQ ID NO. 2:
MGKHVEYLEHVFHYFDEDGDGKISPSELRNRLGLMGGEELIELKEAEEAVESLDSDGDGLLGLEDLARLMELGGGEEEKMKDLRMAFEMYDVEGKGFITPKSLKRMLSRLGESRSVDECKVMINHFDLNGDGVLCFEEFRIMMQ。
example 4 establishment of mulberry VIGS System and functional identification of MmCML37
Utilizing Oligo 7 to design a specific primer MmCML37-F1/R1 (the 5' ends of an upstream primer and a downstream primer are respectively added with EcoR I and Kpn I enzyme cutting sites) of mulberry MmCML37 gene to amplify a target interference fragment, constructing a TRV recombinant virus vector, and inoculating mulberry cotyledons by transforming agrobacterium GV3101 (purchased from Shanghai Toshiyashi limited) and injecting; the primers are as follows:
MmCML37-F1:5'-CGGAATTCATGGGCAAGCACGTAGAGT-3'；
MmCML37-R1:5'-GGGGTACCTCACTGCATCATAATCCTGAAC-3'；
according to the existing establishment technology of mulberry VIGS system in the laboratory, recombinant agrobacterium GV3101 is cultured. Selecting mulberry seedlings with good growth condition and consistent growth, slightly scratching the back of the cotyledon of the mulberry seedlings by using a disposable syringe needle, injecting agrobacterium liquid containing recombinant virus vectors, injecting about 10 mu L of the agrobacterium liquid into each cotyledon, diffusing the agrobacterium liquid in the leaves, taking a plant inoculated with pTRV2 empty vector as a negative Control (CK), taking a wild type inoculated with buffer solution as a blank control (WT), and repeating for 3 times by 10 plants. And (3) selecting seedlings with basically consistent morphological growth vigor 30 days after the recombinant virus vector is injected, transplanting the seedlings into a plastic pot with the upper caliber of 20cm, the lower caliber of 10cm and the height of 20cm for seedling culture, wherein the matrix is vermiculite containing calcium with different concentrations. Respectively using a solution containing 0mg/L, 220mg/L, 440mg/L, 660mg/LCaCl2The mulberry seedlings are treated for 8 days by the MS liquid culture medium. Regularly observing growth trend of mulberry seedlings every day, collecting young leaves of mulberry as experimental material, and using aluminiumWrapping with foil paper, quickly adding into liquid nitrogen, and storing in-80 deg.C refrigerator. In the whole treatment process, the illumination intensity, the light period and the humidity of all the mulberry seedlings are kept unchanged. Then, the detection is carried out according to corresponding physiological and biochemical detection instructions (detection kits are provided by Suzhou Keming Biotechnology Co., Ltd.), and all experiments are repeated for 3 times.
(1) Detecting the content of Malondialdehyde (MDA): about 0.1g of the sample was weighed, 1mL of the extract was added, and ice-bath homogenization was performed. 8000g, centrifuging at 4 deg.C for 10min, collecting supernatant, and placing on ice. Sucking 0.3mL of reagent I into a 1.5mL centrifuge tube, adding 0.1mL of sample, and uniformly mixing; keeping the temperature in 95 deg.C water bath for 30min (covering tightly to prevent water loss), cooling in ice bath, centrifuging at 10000g and 25 deg.C for 10 min; 200. mu.L of the supernatant was pipetted into a 96-well plate, and the absorbance at 532nm and 600nm was measured and calculated.
(2) Proline (Pro) content detection: weighing about 0.1g of sample, adding 1mL of extracting solution, and performing ice bath homogenization; then placing in a water bath at 95 ℃ to shake and extract for 10 min; centrifuging at 25 deg.C for 10min at 10000g, collecting supernatant, and cooling to obtain solution to be detected; preheating the microplate reader for more than 30min, adjusting the wavelength to 520nm, and adjusting the distilled water to zero; adding 0.25mL sample, 0.25mL glacial acetic acid and 0.25mL reagent II into a covered sterile EP tube, placing in a water bath at 95 ℃ for 30min (covering tightly to prevent water loss), and oscillating once every 10 min; after cooling, 0.5mL of toluene is added, the mixture is shaken for 30s, the mixture is kept stand for a while, the pigment is transferred into the toluene, 0.2mL of upper solution is absorbed into a 96-well plate, the color is compared at the wavelength of 520nm, and the absorbance A is recorded and calculated.
(3) Detecting the activity of superoxide dismutase (SOD): about 0.1g of the sample was weighed, 1mL of the extract was added, and ice-bath homogenization was performed. Centrifuging at 8000g and 4 deg.C for 10min, collecting supernatant to obtain solution to be tested, and placing on ice; preheating an enzyme-labeling instrument for more than 30min, adjusting the wavelength to 560nm, and adjusting the distilled water to zero; diluting the reagent with distilled water twice, and using the amount of the reagent (diluting the reagent II with distilled water 1: 1); dissolving a bottle of reagent four in 5mL of distilled water (the reagent four is used up within one week after being dissolved), and then diluting the solution by 4 times with distilled water, wherein the amount of the reagent four and the distilled water are diluted by 1: 3; before measurement, the first, third and fourth reagents are put in water bath at 25 ℃ for more than 5 min; sample determination, namely adding reaction reagents into a 96-well plate in sequence according to the sequence of 45 mu L of a first reagent, 2 mu L of a second reagent, 35 mu L of a third reagent, 100 mu L of a fourth reagent and 18 mu L of sample/distilled water; and standing the uniformly mixed sample at room temperature for 30min, measuring the light absorption value A of each tube at 560nm in an enzyme labeling instrument, and calculating.
(4) Peroxidase (POD) activity assay: about 0.1g of the sample was weighed, 1mL of the extract was added, and ice-bath homogenization was performed. Centrifuging at 8000g and 4 deg.C for 10min, collecting supernatant to obtain solution to be tested, and placing on ice; preheating the microplate reader for more than 30min, adjusting the wavelength to 470nm, and adjusting the distilled water to zero; before measurement, reagent one, reagent two and reagent three were mixed in a ratio of 2.6 (mL): 1.5(μ L): 1 (mu L) and preheating at 25 ℃ for more than 10min to be prepared for use; and adding 10 mu L of sample and 190 mu L of working solution into a 96-well plate, uniformly mixing, placing the mixture into a microplate reader for detection, recording the light absorption value A1 at 470nm for 1min and the light absorption value A2 after 2min, and calculating.
(5) Soluble protein (Cpr) activity assay: weighing about 0.1g of sample, adding 1mL of extracting solution, homogenizing in ice bath, centrifuging at 10000rpm and 4 ℃ for 10min, and taking supernatant to obtain to-be-detected solution; preheating a microplate reader for 30min, adjusting the wavelength to 562nm, and adjusting the distilled water to zero; respectively adding 200 μ L of working solution and 4 μ L of distilled water/standard substance/solution to be detected, mixing reagent A and reagent B at a ratio of 50: 1, covering, mixing to obtain working solution, and preheating the working solution in 60 deg.C water bath for 30 min; mixing, placing at 60 deg.C, keeping the temperature for 30min, loading onto 96-well plate, measuring absorbance A at 562nm, and recording as blank tube A, standard tube A, and measuring tube A respectively.
The mulberry calmodulin protein MmCMML 37 gene is processed by VIGS technology, and the physiological and biochemical indexes of the mulberry are changed after the mulberry calmodulin protein MmCMML 37 gene is silenced: MDA is an important detection index of the damage degree of cell membranes, the content of the MDA reflects the degree of peroxidation of plant membrane lipids, and after VIGS silences MmCML37, compared with blank and negative control, the MDA is increased; pro is an important osmotic regulatory substance, the damage of the plant under the stress can be weakened by increasing the content of Pro, and the Pro can be obviously reduced after VIGS silences genes; the SOD activity in the plant leaves is higher, the capacity of eliminating active oxygen is stronger, the damage of the plants under oxidative stress is smaller, and the SOD after VIGS treatment is obviously reduced compared with a blank control and a negative control; the POD in the plant has strong activity, which is beneficial to the elimination of free radicals, and the POD after VIGS treatment has a trend similar to SOD; soluble protein (Cpr) is an important osmotic regulatory substance, and the Cpr treated by VIGS is obviously reduced compared with a blank control and a negative control; therefore, the mulberry calmodulin protein MmCML37 gene can be used as a marker gene for judging whether mulberry is subjected to abiotic stress.
The above description is only a preferred embodiment of the present invention, and the scope of the present invention is not limited thereto, and any simple modifications or equivalent substitutions of the technical solutions that can be obviously obtained by those skilled in the art within the technical scope of the present invention are within the scope of the present invention.
Sequence listing
<110> university of Jiangsu science and technology
<120> mulberry calmodulin protein MmCML37 and application thereof
<141> 2020-11-26
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gatttgagaa tggcttttga gatgtatgat gtggagggaa aagggttcat aacgcccaag 300
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130 135 140
<210> 9
<211> 27
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 9
cggaattcat gggcaagcac gtagagt 27
<210> 10
<211> 30
<212> DNA
<213> Artificial Sequence (Artificial Sequence)
<400> 10
ggggtacctc actgcatcat aatcctgaac 30
Claims (8)
1. A mulberry calmodulin protein MmCML37, wherein the amino acid sequence of the mulberry calmodulin protein MmCML37 is as shown in SEQ ID No.: 2, respectively.
2. A nucleotide sequence is shown as SEQ ID NO.1, wherein the nucleotide sequence encodes the mulberry calmodulin protein MmCMML 37 of claim 1.
3. A vector comprising the nucleotide of claim 2.
4. A genetically engineered bacterium comprising the vector of claim 3 or having the nucleotide of claim 2 integrated into its genome.
5. Use of the mulberry calmodulin protein mmmcml 37, or a gene encoding it, according to claim 1, for the identification of molecular markers of the mulberry's ability to resist calcium stress.
6. A method for improving the calcium stress resistance of a plant, which comprises allowing a mulberry to express the mulberry calmodulin protein MmCML37 of claim 1.
7. A molecular marker for identifying the calcium stress resistance of mulberry, which is a primer pair for specifically recognizing a coding gene of the polypeptide of claim 1; the primer pair is
MmCML37-F:ATGGGCAAGCACGTAGAGT；
MmCML37-R:TCACTGCATCATAATCCTGAAC。
8. Use of the molecular marker of claim 7 for identifying the resistance of mulberry to calcium stress.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114790449A (en) * | 2022-05-11 | 2022-07-26 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Application of calcium-dependent protein kinase gene GhCPK4 in resisting verticillium wilt of plants |
| CN114958865A (en) * | 2022-04-26 | 2022-08-30 | 中国林业科学研究院林业研究所 | Calmodulin PdCaM247 gene for regulating and controlling poplar xylem development and application thereof |
-
2020
- 2020-11-30 CN CN202011372211.XA patent/CN112390868A/en active Pending
Non-Patent Citations (3)
| Title |
|---|
| HOUQING ZENG等: "Involvement of calmodulin and calmodulin-like proteins in plant responses to abiotic stresses", 《FRONTIERS IN PLANT SCIENCE》 * |
| SANDRA S SCHOLZ等: "Calmodulin-like protein CML37 is a positive regulator of ABA during drought stress in Arabidopsis", 《PLANT SIGNALING & BEHAVIOR》 * |
| 佚名: "PREDICTED: Morus notabilis calcium-binding protein CML37 (LOC21385763), mRNA", 《NCBI》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114958865A (en) * | 2022-04-26 | 2022-08-30 | 中国林业科学研究院林业研究所 | Calmodulin PdCaM247 gene for regulating and controlling poplar xylem development and application thereof |
| CN114790449A (en) * | 2022-05-11 | 2022-07-26 | 新疆农业科学院核技术生物技术研究所（新疆维吾尔自治区生物技术研究中心） | Application of calcium-dependent protein kinase gene GhCPK4 in resisting verticillium wilt of plants |
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