CN116491409B - Rice yield increasing method - Google Patents
Rice yield increasing method Download PDFInfo
- Publication number
- CN116491409B CN116491409B CN202310378700.3A CN202310378700A CN116491409B CN 116491409 B CN116491409 B CN 116491409B CN 202310378700 A CN202310378700 A CN 202310378700A CN 116491409 B CN116491409 B CN 116491409B
- Authority
- CN
- China
- Prior art keywords
- rice
- calcium
- potassium
- osk2
- yield
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Active
Links
- 235000007164 Oryza sativa Nutrition 0.000 title claims abstract description 109
- 235000009566 rice Nutrition 0.000 title claims abstract description 109
- 238000000034 method Methods 0.000 title claims abstract description 12
- 240000007594 Oryza sativa Species 0.000 title 1
- 241000209094 Oryza Species 0.000 claims abstract description 108
- 238000005507 spraying Methods 0.000 claims abstract description 13
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 claims abstract description 10
- 229910001424 calcium ion Inorganic materials 0.000 claims abstract description 10
- 241000196324 Embryophyta Species 0.000 claims description 10
- UXVMQQNJUSDDNG-UHFFFAOYSA-L Calcium chloride Chemical compound [Cl-].[Cl-].[Ca+2] UXVMQQNJUSDDNG-UHFFFAOYSA-L 0.000 claims description 6
- 239000011575 calcium Substances 0.000 abstract description 103
- 229910052791 calcium Inorganic materials 0.000 abstract description 64
- 230000000694 effects Effects 0.000 abstract description 39
- 229910052700 potassium Inorganic materials 0.000 abstract description 39
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 abstract description 38
- 239000011591 potassium Substances 0.000 abstract description 38
- 238000010521 absorption reaction Methods 0.000 abstract description 35
- 239000011148 porous material Substances 0.000 abstract description 16
- 230000000243 photosynthetic effect Effects 0.000 abstract description 11
- NPYPAHLBTDXSSS-UHFFFAOYSA-N Potassium ion Chemical compound [K+] NPYPAHLBTDXSSS-UHFFFAOYSA-N 0.000 abstract description 8
- 229910001414 potassium ion Inorganic materials 0.000 abstract description 8
- 229920000945 Amylopectin Polymers 0.000 abstract description 7
- 230000029553 photosynthesis Effects 0.000 abstract description 6
- 238000010672 photosynthesis Methods 0.000 abstract description 6
- 230000033228 biological regulation Effects 0.000 abstract description 5
- 230000007246 mechanism Effects 0.000 abstract description 5
- 238000004519 manufacturing process Methods 0.000 abstract description 4
- 230000002349 favourable effect Effects 0.000 abstract 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 63
- 210000004027 cell Anatomy 0.000 description 22
- 239000000243 solution Substances 0.000 description 16
- 108091006146 Channels Proteins 0.000 description 13
- 230000001105 regulatory effect Effects 0.000 description 13
- 101001046988 Orthochirus scrobiculosus Potassium channel toxin alpha-KTx 3.7 Proteins 0.000 description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 11
- 230000001276 controlling effect Effects 0.000 description 9
- 238000010586 diagram Methods 0.000 description 9
- 235000013601 eggs Nutrition 0.000 description 9
- 210000002615 epidermis Anatomy 0.000 description 9
- 150000001413 amino acids Chemical class 0.000 description 7
- 238000001514 detection method Methods 0.000 description 6
- 229920000856 Amylose Polymers 0.000 description 5
- 238000005516 engineering process Methods 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 4
- 229920002472 Starch Polymers 0.000 description 4
- 241000746966 Zizania Species 0.000 description 4
- 235000002636 Zizania aquatica Nutrition 0.000 description 4
- 239000003337 fertilizer Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000004044 response Effects 0.000 description 4
- 235000019698 starch Nutrition 0.000 description 4
- 239000008107 starch Substances 0.000 description 4
- 240000008042 Zea mays Species 0.000 description 3
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 3
- 235000013339 cereals Nutrition 0.000 description 3
- 230000008641 drought stress Effects 0.000 description 3
- PGSADBUBUOPOJS-UHFFFAOYSA-N neutral red Chemical compound Cl.C1=C(C)C(N)=CC2=NC3=CC(N(C)C)=CC=C3N=C21 PGSADBUBUOPOJS-UHFFFAOYSA-N 0.000 description 3
- 108090000623 proteins and genes Proteins 0.000 description 3
- 239000002689 soil Substances 0.000 description 3
- 241000894007 species Species 0.000 description 3
- 238000010186 staining Methods 0.000 description 3
- 230000015378 stomatal closure Effects 0.000 description 3
- 241000219194 Arabidopsis Species 0.000 description 2
- 241000219195 Arabidopsis thaliana Species 0.000 description 2
- 239000002028 Biomass Substances 0.000 description 2
- 244000241257 Cucumis melo Species 0.000 description 2
- 235000015510 Cucumis melo subsp melo Nutrition 0.000 description 2
- 102000004257 Potassium Channel Human genes 0.000 description 2
- 241000269368 Xenopus laevis Species 0.000 description 2
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 2
- FJJCIZWZNKZHII-UHFFFAOYSA-N [4,6-bis(cyanoamino)-1,3,5-triazin-2-yl]cyanamide Chemical compound N#CNC1=NC(NC#N)=NC(NC#N)=N1 FJJCIZWZNKZHII-UHFFFAOYSA-N 0.000 description 2
- 239000002253 acid Substances 0.000 description 2
- 239000003513 alkali Substances 0.000 description 2
- 230000036755 cellular response Effects 0.000 description 2
- 235000005822 corn Nutrition 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 108020001213 potassium channel Proteins 0.000 description 2
- 230000001737 promoting effect Effects 0.000 description 2
- 230000008844 regulatory mechanism Effects 0.000 description 2
- 239000011734 sodium Substances 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- 230000035882 stress Effects 0.000 description 2
- 230000005068 transpiration Effects 0.000 description 2
- BJEPYKJPYRNKOW-REOHCLBHSA-N (S)-malic acid Chemical compound OC(=O)[C@@H](O)CC(O)=O BJEPYKJPYRNKOW-REOHCLBHSA-N 0.000 description 1
- 101710172891 ABC transporter G family member 5 Proteins 0.000 description 1
- 241000586291 Ammopiptanthus mongolicus Species 0.000 description 1
- 101100533570 Arabidopsis thaliana SIRK gene Proteins 0.000 description 1
- 238000010356 CRISPR-Cas9 genome editing Methods 0.000 description 1
- 235000008733 Citrus aurantifolia Nutrition 0.000 description 1
- YAHZABJORDUQGO-NQXXGFSBSA-N D-ribulose 1,5-bisphosphate Chemical compound OP(=O)(O)OC[C@@H](O)[C@@H](O)C(=O)COP(O)(O)=O YAHZABJORDUQGO-NQXXGFSBSA-N 0.000 description 1
- 108091006027 G proteins Proteins 0.000 description 1
- 102000030782 GTP binding Human genes 0.000 description 1
- 108091000058 GTP-Binding Proteins 0.000 description 1
- 241000238631 Hexapoda Species 0.000 description 1
- 101001078093 Homo sapiens Reticulocalbin-1 Proteins 0.000 description 1
- 241001100932 Ilex pubescens Species 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102100025335 Reticulocalbin-1 Human genes 0.000 description 1
- 239000004115 Sodium Silicate Substances 0.000 description 1
- 235000011941 Tilia x europaea Nutrition 0.000 description 1
- 235000009754 Vitis X bourquina Nutrition 0.000 description 1
- 235000012333 Vitis X labruscana Nutrition 0.000 description 1
- 240000006365 Vitis vinifera Species 0.000 description 1
- 235000014787 Vitis vinifera Nutrition 0.000 description 1
- 241000269370 Xenopus <genus> Species 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 235000016383 Zea mays subsp huehuetenangensis Nutrition 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 238000012271 agricultural production Methods 0.000 description 1
- 230000009418 agronomic effect Effects 0.000 description 1
- 125000003275 alpha amino acid group Chemical group 0.000 description 1
- BJEPYKJPYRNKOW-UHFFFAOYSA-N alpha-hydroxysuccinic acid Natural products OC(=O)C(O)CC(O)=O BJEPYKJPYRNKOW-UHFFFAOYSA-N 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000021028 berry Nutrition 0.000 description 1
- 230000004790 biotic stress Effects 0.000 description 1
- 239000001110 calcium chloride Substances 0.000 description 1
- 229910001628 calcium chloride Inorganic materials 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000010367 cloning Methods 0.000 description 1
- 230000008094 contradictory effect Effects 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000013613 expression plasmid Substances 0.000 description 1
- 239000013604 expression vector Substances 0.000 description 1
- 230000014509 gene expression Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 238000003306 harvesting Methods 0.000 description 1
- 239000000416 hydrocolloid Substances 0.000 description 1
- 238000005286 illumination Methods 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 229910001410 inorganic ion Inorganic materials 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000004571 lime Substances 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 235000009973 maize Nutrition 0.000 description 1
- 239000001630 malic acid Substances 0.000 description 1
- 235000011090 malic acid Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000035772 mutation Effects 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000014075 nitrogen utilization Effects 0.000 description 1
- 210000000287 oocyte Anatomy 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 230000002018 overexpression Effects 0.000 description 1
- 230000010412 perfusion Effects 0.000 description 1
- 230000035699 permeability Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 230000037039 plant physiology Effects 0.000 description 1
- 239000013612 plasmid Substances 0.000 description 1
- 238000002360 preparation method Methods 0.000 description 1
- 108090000765 processed proteins & peptides Proteins 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000003259 recombinant expression Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000025469 response to water deprivation Effects 0.000 description 1
- 102200016471 rs104894280 Human genes 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000002864 sequence alignment Methods 0.000 description 1
- 238000012163 sequencing technique Methods 0.000 description 1
- 230000011664 signaling Effects 0.000 description 1
- MKWYFZFMAMBPQK-UHFFFAOYSA-J sodium feredetate Chemical compound [Na+].[Fe+3].[O-]C(=O)CN(CC([O-])=O)CCN(CC([O-])=O)CC([O-])=O MKWYFZFMAMBPQK-UHFFFAOYSA-J 0.000 description 1
- NTHWMYGWWRZVTN-UHFFFAOYSA-N sodium silicate Chemical compound [Na+].[Na+].[O-][Si]([O-])=O NTHWMYGWWRZVTN-UHFFFAOYSA-N 0.000 description 1
- 229910052911 sodium silicate Inorganic materials 0.000 description 1
- ZNJHFNUEQDVFCJ-UHFFFAOYSA-M sodium;2-[4-(2-hydroxyethyl)piperazin-1-yl]ethanesulfonic acid;hydroxide Chemical compound [OH-].[Na+].OCCN1CCN(CCS(O)(=O)=O)CC1 ZNJHFNUEQDVFCJ-UHFFFAOYSA-M 0.000 description 1
- 239000007921 spray Substances 0.000 description 1
- 239000012192 staining solution Substances 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 230000002195 synergetic effect Effects 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 230000001960 triggered effect Effects 0.000 description 1
Classifications
-
- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/12—Processes for modifying agronomic input traits, e.g. crop yield
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/10—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in agriculture
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Botany (AREA)
- Developmental Biology & Embryology (AREA)
- Environmental Sciences (AREA)
- Cereal-Derived Products (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The application provides a rice yield increasing method, which comprises the steps of spraying a calcium ion solution to rice leaves every 6 days from a booting stage of rice, and spraying for 6 times to improve the rice yield; the application finds a mode (high-calcium environment) favorable for potassium absorption activity of the rice guard cell potassium ion absorption channel from the regulation and control mechanism of the rice guard cell potassium ion absorption channel, and promotes the rice guard cell potassium ion absorption channel to have lower physiological K + The efficient opening of the leaf pores of the rice under the concentration can promote the photosynthesis of the rice, enhance the photosynthetic efficiency, further improve the production potential and yield of the rice, improve the amylopectin content of the rice and improve the rice quality, and the method is simple and easy to operate and is easy to popularize and apply in fields.
Description
Technical Field
The invention belongs to the technical field of biological agriculture, and in particular relates to a method for regulating and controlling rice yield by improving rice stomata opening efficiency.
Background
With the continuous increase of world population, the continuous reduction of available cultivated land, the continuous occurrence of frequent disaster climate and the like pose a great threat to grain safety, and the utilization of limited cultivated land to further improve grain productivity is an important way for solving the problem. Rice is one of the important food crops, providing energy and nutrition for more than half of the world population. Further improvement of photosynthesis is seen as an optimal way to increase the yield potential of crops, including rice.
The stomata are micropores surrounded by a pair of specialized epidermis cells, namely guard cells, of the epidermis of the plant leaf, and are used for carrying out water transpiration and obtaining photosynthetic CO by the plant 2 The main portal for substrates. The organic or inorganic ions in the stomatal guard cells include K + The concentration of malic acid, saccharides and the like can be changed dynamically to control the movement of guard cells and further the opening degree of air holes. Current research is generally focused on plant drought stress response to reduce stomatal closure due to water loss, and stomatal immunity triggered when plants are invaded by stomata such as pathogenic bacteria, which is when a large amount of calcium ions enter guard cells around the stomata to trigger stomatal closure (berry, m., lipp, P).&Bootman,M.The versatility and universality ofcalcium signalling. NatRevMol Cell, biol.2000,1, 11-21), which are necessary for crops susceptible to drought or biotic stress, intensive mechanism studies are currently focused mainly on the model plant Arabidopsis.
Under drought stress, rice ATP binding cassette subfamily G protein RCN1/OsABCG5 is involved in the accumulation of ABA in guard cells and is an indispensable component for stomatal closure. For crops with very superior water conditions such as rice, the requirement of closing the stomata in response to drought stress is relatively not urgent, if the efficient open state of the stomata can be promoted, the root system can be pulled to absorb and upwards transmit soil nutrients, the utilization efficiency of the nutrients is improved, and meanwhile, the soil nutrients and the high photosynthetic CO caused by the stomata can be pulled 2 The assimilation forms C, N, K with high efficiency and synergistic effect, and the crop yield is further improved. Previous researches show that the stomatal opening efficiency is closely related to the activity of the guard cell stomatal opening potassium ion channel, and the overexpression of the stomatal opening potassium ion absorption channel OsK-1 can improve the photosynthetic rate, the stomatal permeability, the transpiration rate, the nitrogen utilization efficiency and the like of rice (Chinese patent ZL 201810959909.8). Under normal physiological state of rice, its apoplast K + The concentration is generally below 10mM, which protects the low concentration of K around the cells + The environment inhibits the opening of the stomata of the rice, so that the improvement of the yield of the rice is restricted, and the opening efficiency of the stomata is closely related to the activity of the potassium ion absorption channel of the stomata of the guard cells. In addition, the potassium fertilizer is added in production, so that the potassium fertilizer can be added to improve the potassium nutrition of crops, the yield of the crops can be effectively improved, the tolerance of the crops to adverse growth conditions such as drought, salt damage, plant diseases and insect pests, lodging and the like can be enhanced, but the approach needs to supplement a large amount of potassium fertilizer, which is contradictory to the phenomenon of the existing potassium fertilizer resource shortage.
Currently in agricultural production, ca + Is generally applied to the inhibition of plant production under the saline-alkali or acid condition, wherein lime CaO is applied to the soil to neutralize the pH value to reduce acid toxicity or reduce saline-alkali stress and the like. While Ca + The application of promoting the stomata opening of rice leaves and improving photosynthesis has not been reported yet.
Disclosure of Invention
Aiming at the problems, the invention aims to provide a convenient field operation technology for promoting the yield increase of rice, which finds a mode (high calcium) beneficial to the potassium absorption activity of the rice guard cells from the regulation and control mechanism of potassium ion absorption channels of the rice guard cells and promotes the potassium absorption activity of the rice guard cells to be in a lower physiological K + The efficient opening of the leaf pores of the rice under the concentration promotes the photosynthesis of the rice, namely, the CaCl is directly sprayed on the leaf surface 2 The effect of increasing the yield by 15% can be achieved in the field.
Specifically, the invention is realized by the following technical scheme:
a rice yield increasing method comprises the following specific steps: spraying a calcium ion solution to rice leaves every 6 days from the booting period (growing period) of the rice for 6 times; so as to regulate and control the opening of the rice stomata, promote the efficient opening of the rice leaf stomata, enhance the photosynthetic efficiency and further improve the rice yield and the amylopectin content of the rice.
Further, the concentration of calcium ions in the above-mentioned calcium ion solution was 20mM.
Further, the ionic solution is preferably a calcium chloride solution, and the spraying amount is 4 ml/plant of rice.
First, in one embodiment of the present application, the activity of the epidermal guard cells was verified by scraping the rice subleaf epidermis, neutral red, and immersing the active epidermis in the following solution: 5mM K + +0.05mM Ca 2+ ,5mM K + +5mM Ca 2+ After half an hour of illumination, the pore opening degree of the rice is detected by an optical microscope, and the result shows that the rice is mixed with low calcium 5mM K in wild rice Japanese sunny + +0.05mM Ca 2+ Compared with the condition, the pore opening degree of the rice is 5mM K higher than that of the calcium + +5mM Ca 2+ The air hole opening is obviously promoted under the condition that the average increase of the air hole opening is 10 percent; previous studies showed that OsK2-1 (belonging to the fundamental channel of rice) was closely related to rice stomatal opening, and that the applicant further created OsK2-1 gene deleted mutant osk-1, which showed that under high calcium conditions, the stomatal opening of osk2-1 mutant rice lost the effect of being promoted by high calcium, showing no significant difference between stomatal opening at low and high calcium.
Second, according to the study of electrophysiological technology, the potassium absorption function of OsK2-1 is obviously promoted by extracellular high calcium, and the promotion phenomenon is closely related to the 96 th amino acid D of a specific site of OsK2-1, compared with homologous channels of Arabidopsis thaliana, corn, common melon and drought-resistant plant ammopiptanthus mongolicus, the SNP site is unique to rice OsK-1, and if the site D is mutated into N corresponding to homologous channels of other species, the potassium absorption activity of rice OsK2-1 is lost by the effect of high calcium promotion.
Thirdly, one example of the present application demonstrates that the effect of potassium absorption activity of OsK-1 promoted by high calcium is caused by extracellular calcium, this step being achieved by: at K + Under the condition of constant concentration, frog eggs expressing OsK2-1 are sequentially irrigated with 1.8mM Ca by a double-electrode voltage clamp technology 2+ -10mM Ca 2+ -1.8mM Ca 2+ The solution was recorded for 15 minutes and frog eggs injected with water were used as controls. By varying the extracellular calcium concentration, the results indicate that when extracellular calcium is increased, the potassium uptake capacity of OsK2-1 is increased and the calcium-promoting effect of the OsK2-1 potassium uptake activity is caused by extracellular calcium. However, the calcium promotion phenomenon of potassium absorption of OsK 2-1D 96N after D96 in OsK-1 is changed into N is eliminated, and the existence of D96 of rice OsK2-1 is closely related to the calcium promotion phenomenon. The phenomenon that the opening degree of the rice air holes is promoted by high calcium is closely related to the phenomenon that the potassium absorption activity of OsK2-1 is promoted by high calcium, and the 96 th amino acid D is a key site of the rice air holes promoted by calcium.
Further electrophysiological researches find that the rice stomatal guard cells have two potassium ion potassium channels OsK-1 and OsK2-3, wherein OsK2-3 has no potassium absorption function, so that the phenomenon that the potassium absorption activity is regulated and controlled by calcium does not exist, osK2-1 has potassium absorption activity and is strongly promoted by external high calcium, the increase of intracellular calcium cannot promote the potassium absorption activity, the promotion phenomenon is closely related to the D96 amino acid site of OsK2-1 protein, and the promotion phenomenon disappears after the D96 site is mutated into other amino acid N corresponding to channels with the potassium absorption activity not promoted by Ca; on the contrary, N of the corresponding sites of the two (corn and arabidopsis) stomatal potassium absorption channels without a calcium promotion mechanism is changed into D of rice, and the result shows that the two sites all have the calcium promotion phenomenon, which indicates that the special variation of D96 amino acid leads OsK2-1 to have a special regulation mechanism of calcium promotion, and the promotion mechanism is special for rice.
Therefore, based on the regulation mechanism that the potassium absorption activity of the pore guard cell potassium ion absorption channel on the epidermis of the rice leaf is strongly promoted by external high calcium, the application firstly sprays the calcium ion solution to the rice leaf from the booting stage (growth period) of the rice so as to regulate and control the opening degree of the pore of the rice, promote the efficient opening of the pore of the rice leaf, enhance the photosynthetic efficiency and further improve the production potential and yield of the rice, improve the amylose content of the rice and improve the quality of the rice.
Drawings
FIG. 1 is a chart showing pore opening staining and statistics of wild type rice and osk2-1 mutant rice at different calcium concentrations;
wherein A, B is a neutral red staining chart of subleaf epidermis scraped by Japanese wild rice and osk2-1 mutant (red cells indicate that the cells are in active state),
c is low calcium 0.05mM Ca 2+ And high calcium 5mM Ca 2+ Pore opening degree of wild type rice Nippon (NB) and osk2-1 mutant rice.
FIG. 2 is a schematic diagram showing the results of a test for the potassium absorption activity of OsK2-1 promoted by calcium and the potassium absorption activity of SNP locus OsK-1D 96N thereof inhibited by calcium;
wherein A is OsK2-1 calcium response SNP locus analysis chart, B is OsK2-1 in different Ca 2+ The potassium absorption activity current at the concentration is shown in a schematic way, C is OsK-1, and the concentration is different in Ca 2+ The current-voltage relationship of potassium absorption activity under concentration, D is OsK2-1 at different Ca 2+ Statistical graph of potassium absorption activity current at concentration, E is OsK 2-1D 96N at different Ca 2+ The potassium absorption activity current at the concentration is shown in a schematic way, F is OsK 2-1D 96N at different Ca 2+ Current-voltage relationship of potassium-adsorbing activity at concentration, G is OsK 2-1D 96N at different Ca 2+ Concentration of potassium uptake activity current statistics.
FIG. 3 is a graph showing the results of an extracellular calcium-promoted assay for potassium absorption activity of OsK 2-1;
wherein A is a schematic diagram of the influence of extracellular calcium on OsK2-1 potassium absorption activity, and B is a statistical diagram of the influence of extracellular calcium on OsK2-1 potassium absorption activity.
FIG. 4 is a field blade Ca 2+ Regulating and controlling (spraying calcium chloride solution on leaves) the photosynthesis performance and agronomic index result schematic diagram of the rice;
wherein A is Ca 2+ The photosynthetic rate detection result of the regulated rice is shown in the diagram, and B is Ca 2+ Results of detecting the air pore conductivity of rice are shown in the figure, C is Ca 2+ Schematic diagram of detection result of regulating and controlling biomass of overground parts of rice, wherein D is Ca 2+ And (5) regulating and controlling rice yield detection results.
FIG. 5 is a schematic diagram showing the effect of foliar calcium regulation on starch content in rice;
wherein A is Ca 2+ The detection result of regulating and controlling the total starch content of the rice is shown in a schematic diagram, and B is Ca 2+ The detection result of regulating and controlling the amylose content of rice is shown in the schematic diagram, C is Ca 2+ And regulating and controlling the amylopectin content detection result of the rice.
Detailed Description
The following technical solutions of the present application are further described with reference to examples:
the rice varieties referred to in the examples are common varieties in which Nippon rice is obtained by giving away from the university of Nanjing student's college of life.
osk2-1 mutant was obtained by CRISPR-Cas9 technology from Wuhan Bo remote biotechnology Co.
The culture medium referred in the examples:
the international rice nutrient solution comprises: 1.25mM NH 4 NO 3 ,0.3mM KH 2 PO 4 ,0.35mM K 2 SO 4 ,1mM CaCl 2 ·2H 2 O,1mM MgSO 4 ·7H 2 O,0.5mM Na 2 SiO 3 ,20μM NaFeEDTA,20μM H 3 BO 3 ,9μM MnCl 2 ·4H 2 O,0.32μM CuSO 4 ·5H 2 O,0.77μM ZnSO 4 ·7H 2 O,0.39μM Na 2 MoO 4 ·2H 2 O, the balance is water and the pH is 5.8.
Example 1 Effect of different calcium concentrations on pore opening of Nippon Rice and osk2-1 mutant Rice
In this example, wild rice seedlings (Nippon Temminck) and osk-1 mutants (Nippon Rice with OsK-2-1 gene knocked out) were cultured in International Rice nutrient solution containing no sodium silicate for 7 days, then the upper epidermis and mesophyll were gently scraped off with a sharp blade, and the scraped lower epidermis was placed in 0.1mg/mL neutral red (live cell stain) staining solution for 10 minutes, and in FIG. 1, A, B was the staining results of wild type and osk2-1 mutant lower epidermis, respectively, and it was seen that both cells were in an active state. The two rice hypodermis were then separately placed in low calcium (5 mM K) + +0.05mM Ca 2+ ) Or high calcium (5 mM K) + +5mM Ca 2+ ) In the solution, the air hole opening was counted by photographing under a high power microscope and imaging under light for 30 minutes, and the result is shown in fig. 1C.
The results showed that the calcium-rich peptide was low in calcium (0.05 mM K + +5mM Ca 2+ ) Compared with the condition, the pore opening degree of the wild rice is high in calcium (5 mM K + +5mM Ca 2+ ) The method has the advantages that the method is obviously promoted under the condition that the average increase of the air hole opening is 10%; the osk-1 mutant rice loses the effect of being promoted by high calcium, and shows that the pore opening has no obvious difference under low calcium and high calcium.
Example 2OsK2-1 and its calcium response site D96N Potassium-adsorbing Activity is promoted by calcium
By sequence search, the applicant has aligned OsK-1 and its homologous channels in other species such as Arabidopsis thaliana (AtKAT 1: NP-199436, atKAT2: NP-193563), maize (ZmK 2.1: AJ 132686), grape (SIRK: RVW 16660.1), melon (MIRK: NP-001284388.1) and the drought-resistant plant ilex pubescens (AmKAT 1: KP 772269), and the amino acid sequence alignment found OsK-1 that at amino acid D at position 96, unlike other homologous channels, has site specificity as shown in FIG. 2A.
Mutation of D at position 96 of OsK2-1 to N by overlap PCR, followed by cloning OsK-1 and OsK 2-1D 96N, ligation to animal expression vector pCI, and ligationThe correct recombinant expression plasmid containing OsK2-1 or OsK 2-1D 96N was obtained by over-sequencing. The recombinant plasmids were separately injected into mature frog egg cells (wherein Xenopus laevis was purchased from Shanghai Stem cell institute, frog egg cell reference V ary, a. -a., bosseux, c., gaymard, f., sentenac, h., thibaud, j. -b.level of expression in Xenopus oocytes affects some characteristics of a plant inward-rectifying voltage-gate K.) + Pfliiger Arch,1994,428:422-424, from Xenopus laevis), for about 48 hours at 21 ℃. Sequential perfusion of 0mM Ca 2+ ，1.8mM Ca 2+ 10mM Ca 2+ The solution is subjected to functional research by a double-electrode voltage clamp technology.
The results indicate that K of OsK2-1 + The absorption capacity is significantly promoted by calcium ions (shown as B in fig. 2). Further current versus voltage (shown as C in FIG. 2) and statistics (shown as D in FIG. 2) also demonstrate K of OsK2-1 as calcium ion concentration increases + The absorption capacity is significantly increased. On the contrary, after the 96 th amino acid D of OsK-1 is mutated into the corresponding N in the homologous channel of other species sources, the calcium promotion effect of potassium absorption is eliminated, and even a certain degree of high calcium inhibition phenomenon (shown as E-G in fig. 2) occurs, further showing that the OsK2-1 unique calcium promotion phenomenon is unique to rice.
Example 3OsK2-1 Potassium absorption Activity is promoted by extracellular calcium
This example investigated the effect of extracellular calcium on OsK-1 channel activity (long-term record). Extracellular calcium: 1.8mM Ca was sequentially perfused 2+ -10mM Ca 2+ -1.8mM Ca 2+ Solution (solution basic composition: 50mM KCl, 1mM MgCl) 2 、X mM CaCl 2 Y mM NaCl,5mM HEPES-NaOH, pH 7.4, caCl 2 And NaCl total concentration 50mM, adjusted according to the experimental requirement), using frog eggs injected with water as control. The results showed that when extracellular calcium was increased from 1.8mM to 10mM, the current level of the control frog egg injected with water was not affected by the change in extracellular calcium concentration, whereas the potassium uptake current of the frog egg injected with OsK2-1 increased sharply and tended to stabilize, and when extracellular calcium eluted from 10mM back to 1.8mM, the potassium uptake current of OsK2-1 was also restored to the starting waterFlat (stable response record), FIG. 3A is a current schematic of single frog egg cell response to extracellular calcium, and FIG. 3B is a current statistical graph of frog egg cell response to extracellular calcium by injecting OsK 2-1. This result demonstrates that altering the concentration of extracellular calcium ions, osK2-1, increases the potassium uptake capacity and that the increase in extracellular calcium promotes the potassium uptake function of OsK 2-1.
EXAMPLE 4 blade Ca 2+ Regulating and controlling (blade spraying calcium solution) to enhance pore conductance and photosynthetic rate of rice
The calcium preparation used in this example was 20mM CaCl 2 Acting on rice leaves (cultivar Nanjing 46, commercially available variety), starting spraying at booting stage of rice, spraying once every 6 days for 6 times, wherein each spraying amount is about 4 ml of each plant of rice, and the concentration is 20mM CaCl 2 Solution (Ca) 2+ Regulation) while setting up the sprayed water as a control group (control).
The photosynthetic rate and pore conductance were measured by the methods of reference (Makino, a., mae, t.and Ohira, k.photosynthesis and Ribulose1,5-Bisphosphate Carboxylase in Rice leavs.plant Physiology,1983,73,1002-1007), and the results showed that the photosynthetic rate (shown in fig. 4 a) and pore conductance (shown in fig. 4B) of the calcium solution-sprayed rice were significantly improved by 10% and 44%, respectively, over the control-sprayed water. In the harvest period, the field yield measurement result shows that compared with water spraying, the biomass of the overground parts of the rice (shown as C in figure 4) and the yield of the rice (shown as D in figure 4) sprayed with calcium are obviously increased, and the increase is 17% and 15% respectively.
EXAMPLE 5 blade Ca 2+ Regulating and controlling (spraying calcium chloride solution on leaves) to improve taste quality of rice
The amylopectin content is an important index for determining the taste quality of rice, and in general, rice having a high taste quality has a high amylopectin content. This example shows the calcium-regulated rice of example 4 using the method disclosed in the literature "Dawei Zhu, changyun Fang, zihui Qian et al Differences in starch structure, physicochemical properties and texture characteristics in superior and inferior grains of rice varieties with different amylose contents food Hydrocolloids,2021The results of the amylose and amylopectin content in the rice show that compared with the control group (control) rice sprayed with water, the control group (Ca) of the calcium chloride ion solution is sprayed 2+ regulation) rice has an increased total starch content (shown as a in fig. 5), an approximately 2-fold decrease in amylose content (shown as B in fig. 5), and an approximately 80% increase in amylopectin content (shown as C in fig. 5). The method shows that the photosynthetic performance of the rice is regulated and controlled by calcium, so that not only can the productivity of the rice be obviously increased, but also the taste quality of the rice can be improved.
Claims (1)
1. The rice yield increasing method is characterized by comprising the following specific steps: spraying a calcium chloride solution to rice leaves every 6 days from the booting stage of the rice for 6 times so as to improve the yield of the rice; the concentration of calcium ions in the calcium chloride solution is 20mM, and the spraying amount is 4 milliliters per plant of rice.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310378700.3A CN116491409B (en) | 2023-04-11 | 2023-04-11 | Rice yield increasing method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310378700.3A CN116491409B (en) | 2023-04-11 | 2023-04-11 | Rice yield increasing method |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116491409A CN116491409A (en) | 2023-07-28 |
| CN116491409B true CN116491409B (en) | 2024-01-23 |
Family
ID=87315894
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310378700.3A Active CN116491409B (en) | 2023-04-11 | 2023-04-11 | Rice yield increasing method |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116491409B (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001192310A (en) * | 2000-01-12 | 2001-07-17 | Mitsui Chemicals Inc | Method for promoting absorption of calcium ion from surface of plant |
| KR100495933B1 (en) * | 2004-10-07 | 2005-06-16 | 장권일 | Rice having high quantity calcium and methods of cultivating the rice |
| CN108358693A (en) * | 2018-02-28 | 2018-08-03 | 中国科学院南京土壤研究所 | The foliar fertilizer and its preparation method and application of calcium content in a kind of raising rice grain |
| CN108675891A (en) * | 2018-08-29 | 2018-10-19 | 湖南农业大学 | A kind of new foliar fertilizer |
| CN108835124A (en) * | 2018-05-30 | 2018-11-20 | 东北农业大学 | A kind of cold-resistance agent that rice is influenced for mitigating boot stage chilling injury and its application |
| CN109082428A (en) * | 2018-08-22 | 2018-12-25 | 中国科学院南京土壤研究所 | The application of rice stomatal opening type potassium-channel gene OsK2-1 and its expression vector |
-
2023
- 2023-04-11 CN CN202310378700.3A patent/CN116491409B/en active Active
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2001192310A (en) * | 2000-01-12 | 2001-07-17 | Mitsui Chemicals Inc | Method for promoting absorption of calcium ion from surface of plant |
| KR100495933B1 (en) * | 2004-10-07 | 2005-06-16 | 장권일 | Rice having high quantity calcium and methods of cultivating the rice |
| CN108358693A (en) * | 2018-02-28 | 2018-08-03 | 中国科学院南京土壤研究所 | The foliar fertilizer and its preparation method and application of calcium content in a kind of raising rice grain |
| CN108835124A (en) * | 2018-05-30 | 2018-11-20 | 东北农业大学 | A kind of cold-resistance agent that rice is influenced for mitigating boot stage chilling injury and its application |
| CN109082428A (en) * | 2018-08-22 | 2018-12-25 | 中国科学院南京土壤研究所 | The application of rice stomatal opening type potassium-channel gene OsK2-1 and its expression vector |
| CN108675891A (en) * | 2018-08-29 | 2018-10-19 | 湖南农业大学 | A kind of new foliar fertilizer |
Non-Patent Citations (3)
| Title |
|---|
| 外源Ca2+对PEG处理下转C4型PEPC基因水稻光合生理的调节;刘小龙等;《植物学报》;第50卷(第02期);第206-216页 * |
| 涂仕华.《常用肥料使用手册（修订版）》.四川科学技术出版社,2014,(第02版),第107-108页. * |
| 钙对水稻苗期光合特性影响及其亚细胞分布;朱国奇等;《中国农学通报》;第31卷(第12期);第92-98页 * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116491409A (en) | 2023-07-28 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Wallace | Growth, morphology and gas exchange of mycorrhizal and nonmycorrhizal Panicum coloratum L., a C 4 grass species, under different clipping and fertilization regimes | |
| CN102599040B (en) | Soilless blueberry cultivation method | |
| WO2007029775A1 (en) | Low light cultivation method and plant growth promoting agent | |
| TW201433264A (en) | Method of increasing abiotic stress resistance of a plant | |
| Ho | The physiological basis for improving tomato fruit quality | |
| CN110463545A (en) | A kind of irrigation method of the antifouling high yield of rice water-saving emission reduction | |
| FI98512C (en) | Improving crop yield in plants | |
| CN102960151A (en) | Method for increasing low-temperature resistance of rice seedling stage | |
| CN116491409B (en) | Rice yield increasing method | |
| Yuan et al. | Reduced square Bacillus thuringiensis insecticidal protein content of transgenic cotton under N deficit | |
| Mennan et al. | The effects of depth and duration of seed burial on viability, dormancy, germination, and emergence of ivyleaf speedwell (Veronica hederifolia) | |
| HUT77910A (en) | The exogenous use of betaine to improve the yield of c-4 cereals, method based on this use and c-4 cereals treated by this method | |
| CN102643856B (en) | Applications of At-ACA8 gene of arabidopsis thaliana for enhancing plant stress resistance and regulating plant growing development | |
| Bader et al. | Effect of water deficit and foliar application of amino acids on growth and yield of eggplant irrigated by two drip systems under greenhouse conditions | |
| WO1997008951A1 (en) | Improving the yield of plants | |
| JPH08157317A (en) | Method for enhancing drought resistance or salt tolerance of plant | |
| Balliu et al. | Nitrogen effects on the relative growth rate and its components of pepper (Capsicum annuum) and eggplant (Solanum melongena) seedlings | |
| Gao et al. | Effect of grafting on cold tolerance in eggplant seedlings | |
| Zhou et al. | Effects of rootstock on fruit yield and quality of hydroponically cultivated grafted cucumber under NaCl stress | |
| CN111011394B (en) | Wheat cold-resistant inducer and application method thereof | |
| CN115232764B (en) | Microbial agent | |
| CN116941639B (en) | Application of sodium nitroprusside in relieving high temperature heat injury in flowering phase of rice | |
| LU501250B1 (en) | Saline land improver for wheat cultivation | |
| CN107119047B (en) | The preparation and application of the codon optimization of engineer's structure and chemical synthesis gene rdh and its expression product polypeptide G1 | |
| Mahlare et al. | Response of Cyclopia Subternata to Watering Frequency: Stomatal Conductance, Proline, and Relative Water Content |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| TR01 | Transfer of patent right |
Effective date of registration: 20240613 Address after: 201400 floor 5, building 11, No. 6055, Jinhai highway, Fengxian District, Shanghai Patentee after: Shanghai Taibo Photosynthetic Agricultural Technology Co.,Ltd. Country or region after: China Address before: 210008 Xuanwu District, Jiangsu, Beijing East Road 71, Nanjing Patentee before: INSTITUTE OF SOIL SCIENCE, CHINESE ACADEMY OF SCIENCES Country or region before: China |