CN115918516A - Method for reducing pistil aberration rate of tobacco sua cytoplasmic male sterile line and breeding and planting - Google Patents
Method for reducing pistil aberration rate of tobacco sua cytoplasmic male sterile line and breeding and planting Download PDFInfo
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Abstract
The application discloses a method for reducing pistil aberration rate and breeding and planting of a tobacco sua cytoplasmic male sterile line, which comprises the following steps: spraying KH on the whole leaf surface of the tobacco plant at the bud emergence prophase of the tobacco plant 2 PO 4 Spraying solution with at least 10 mKH per tobacco 2 PO 4 A solution; spraying KH 2 PO 4 The concentration of the solution is more than or equal to 0.1g/L. The method can obviously reduce the ratio of flower and fruit dropping after pollination caused by pistil deformity in the production of the tobacco sterile line seeds, effectively reduce the economic loss generated in the seed reproduction production, obviously reduce the pistil deformity rate, improve the fruit setting rate and improve the adverse effect of the pistil deformity of the tobacco sua cytoplasmic male sterile line on the seed production quality.
Description
Technical Field
The application relates to the technical field of tobacco seed reproduction, in particular to a method for reducing pistil aberration rate of a tobacco sua cytoplasmic male sterile line and performing seed reproduction and planting.
Background
The tobacco male sterile line and hybrid are widely popularized and applied, and gradually become the main development trend and direction of tobacco production. The method has the advantages of ensuring the purity of the seeds, fully exerting the heterosis of the tobacco male sterile line and improving the quality of the seeds by utilizing the multiple advantages of the seed reproduction of the tobacco male sterile line, and realizing the positioning production of a variety production area according to the requirement of the industry on raw materials.
At present, sterile lines (such as MS K326 and MS Yunyan 87) popularized and planted in China production mainly have cytoplasm from fragrant and sweet tobacco (N. Suaveolens), and the sua cytoplasm sterile line has no adverse effect on field agronomic characters and economic benefits, and is suitable for being used as a female parent to produce F1 seeds.
However, the stamen of the sua cytoplasmic sterile line is various in shape, and serious flower and fruit dropping is often caused once the stamen is subjected to malformation and pollination, so that the waste of high-quality pollen of the male parent is caused, the seed production quality is greatly influenced, adverse effects are brought to the tobacco seed reproduction production, and the tobacco seed reproduction cost is increased.
The pistil form seen in the prior tobacco seed reproduction production is shown as a-g) in figure 1, which is respectively a) a flower column in figure 1 a) is circular, and the stigma is divided into two parts; FIG. 1 b) the style is circular, with trisplit stigma; FIG. 1 c) the style is circular, with multiple chapiters; FIG. 1 d) the style is flat and short; FIG. 1 e) flat and long style; f) Cracking the flower column; g) Separating the flower columns.
Whether all pistils in the shapes shown as a-g) in the figure 1 can cause flower and fruit dropping after pollination of the pistils of the sterile line is not clearly indicated in the prior art, and the technical content of how to take measures to reduce the proportion of deformed pistils which cause flower and fruit dropping after pollination is further not disclosed.
Disclosure of Invention
The application provides a method for reducing pistil aberration rate of a tobacco sua cytoplasmic male sterile line and breeding and planting, which is used for solving the technical problems that serious flower and fruit dropping phenomena exist after pollination of part of morphological pistils in tobacco breeding production in the prior art, the tobacco is seriously reduced in yield and economic loss, and the flower and fruit dropping phenomena after pollination are reduced due to the lack of effective measures.
The application provides a method for reducing pistil aberration rate and breeding and planting of a tobacco sua cytoplasmic male sterile line, which comprises the following steps: spraying KH on the whole leaf surface of the tobacco plant at the bud emergence prophase of the tobacco plant 2 PO 4 Solution of tobacco plantsSpraying at least 10ml KH 2 PO 4 A solution; spraying KH 2 PO 4 The concentration of the solution is more than or equal to 0.1g/L.
According to the method, pistil morphology with flower and fruit dropping after pollination is determined through experiments for the first time, the pistil morphology is improved by the method, the effect is obvious, and pistil aberration rate can be reduced by more than 15%. Spraying KH 2 PO 4 The concentration of the solution is more than or equal to 0.1g/L. Can exert the effect of improving the distortion rate, and the distortion rate is reduced to 55.93 percent in the embodiment.
Spraying KH 2 PO 4 The solvent of the solution is water.
In various forms of pistils of the cytoplasmic male sterile line of the tobacco, only the flat columns become flat, namely the flat columns and the columns break, namely the columns break, so that the fruit setting rate after pollination is reduced and the number of single fruits is reduced, and the round columns and the column head form have no influence on the fruit setting rate and the number of single fruits after pollination. Classifying the flat and short and long of the style into flat and long according to the fruit setting rate and the single fruit grain number level after pollination; splitting and separating the flower column into split columns; the cylindrical stigma of the style is divided into two parts, the cylindrical stigma of the style is divided into 3 parts and the cylindrical stigma of the style is divided into a plurality of parts. Pistils of the oblate and schizont sterile lines, which cause the reduction of the fruit setting rate and the reduction of the number of single fruit grains after pollination, are defined as deformed pistils; the pistil of the cylindrical sterile line which has no influence on the fruit setting rate and the number of single fruit grains after pollination is defined as the normal pistil.
The method is provided on the basis of sufficient and repeated experiments, and through field practice inspection, the method can obviously improve the pistil aberration rate of the sua cytoplasmic male sterile line of the tobacco and improve the fruit setting rate after pollination.
Preferably, KH is sprayed once 2 PO 4 The interval time of the solution is 5-10 days until the tobacco seed reproduction and pollination period is finished.
Preferably, the gynoecium aberration rate of the sua sterile line is reduced by more than 15% after the method is adopted.
Preferably, the distortion ratio is calculated as follows:
pistil distortion rate = (number of malformed pistils/(number of normal pistils + number of malformed pistils)) x100%, where the number of normal pistils means: the flower column is circular, and the column head is divided into two parts; or the flower column is round, and the column head is three-split; or the total number of pistils with round style and multiple chapiters, in particular the total number of pistils in the style shown in figures 1 a) to c);
the number of pistils with malformation refers to: the flower pillar is flat and short; or the flower column is flat and long; or the flower column is cracked; or the sum of the number of pistils separated by the style. In particular to the sum of the number of pistils in the flower-column shape shown in figures 1 d) to g);
in the application, the stigma of the normal pistil has a complete columnar structure, the stigma of the style is only a top end with two cracks, three cracks or multiple cracks, and the stigma is not cracked to the lower part of the style;
malformed pistils refer to: malformed pistils refer to: the flower pillar is flat and short; or the flower column is flat and long; or the flower column is cracked; or pistils isolated from the style;
preferably, the sterile cytoplasm of the sua male sterile line tobacco variety is derived from sweet tobacco (n. Preferably, MS Yunyan 87 and MSK326..
Preferably, KH 2 PO 4 The concentration of the solution is 0.1-5 g/L, and the effect of improving the distortion rate can be normally exerted at the moment.
Preferably, KH 2 PO 4 The concentration of the solution was 1g/L. The pistil aberration rate after spraying this concentration in the examples was only 47.4%.
Preferably, the tobacco plant is planted on tobacco continuous cropping soil.
According to the operation, the problem that the pistil malformation proportion is too high due to the fact that continuous cropping obstacles affect the absorption of potassium elements from continuous cropping tobacco soil of tobacco plants can be effectively solved, the pollination success rate can be effectively improved, the proportion of flower and fruit dropping after tobacco pollination is reduced, and the yield is improved. The continuous cropping soil can be the continuous cropping soil of various tobaccos.
The beneficial effects that this application can produce include:
1) The method for reducing the pistil aberration rate of the tobacco sua cytoplasmic male sterile line can obviously reduce the pistil aberration rate in the production of tobacco sterile line seeds, improve the yield after pollination and effectively reduce the economic loss generated in seed reproduction production.
2) The method for reducing pistil aberration rate and breeding and planting of the cytoplasmic male sterile line of tobacco sua, which is provided by the application, firstly proposes that KH is sprayed on the whole leaf surface of a tobacco plant at the bud early stage of the tobacco plant 2 PO 4 Improving pistil malformation of cytoplasmic male sterile line of tobacco sua, and spraying KH 2 PO 4 The post-tobacco plants are quickly absorbed and take effect quickly, the pistil aberration rate of the sua cytoplasmic male sterile line of the tobacco can be obviously reduced by more than 15 percent, and the fruit setting rate of the pollinated plants is effectively improved. The KH is used 2 PO 4 Is a high-efficiency phosphorus-potassium compound fertilizer commonly used in agriculture, and has high safety and low cost. By using the method, high-quality male parent pollen can be saved, the seed production quality is improved, labor is saved, and the tobacco seed breeding cost is reduced.
3) The planting method provided by the application can effectively improve the pistil aberration rate of the tobacco sua cytoplasmic male sterile line on the continuous cropping tobacco soil.
Drawings
FIG. 1 is a photograph of pistil form in the prior art tobacco seed reproduction, a) the style is circular and the stigma is double-split; b) The flower column is circular, and the column head is three-split; c) The flower column is circular, and the column head is cracked; d) The flower pillar is flat and short; e) The flower pillar is flat and long; f) Cracking the flower column; g) Separating the flower columns;
the application provides a schematic diagram of a method for reducing pistil aberration rate of a tobacco SUA cytoplasmic male sterile line;
FIG. 2 is a fluorescent micrograph of pollen germination and pollen tube elongation in a style after pollination of the various styles of the present application in example 3; wherein a) a photograph of a cylindrical style shows that a large number of pollen normally germinates on the stigma; b) The picture is a cylindrical style picture, and shows that a large number of pollen tubes grow smoothly in the style; c) The picture is a cylindrical style picture, and shows that a large number of pollen tubes smoothly extend into an ovary to germinate; d) The pollen tube is a flat column type flower column, shows that pollen germinates on the column head, and part of the pollen tube extends into the flower column; e) The flower column is flat column type, and shows that part of the pollen tube grows in the flower column; f) The display part of the pollen tube extends into the ovary; g) The pollen is a column-splitting style of style, which shows that part of pollen germinates on the column head, and a small amount of pollen tube extends into the style; h) A split-column style of style shows a small number of pollen tubes growing in the style; i) The pollen tube is a column-split type pollen column, and is shown to be stopped at the junction of the pollen column and the ovary, and a very small amount of pollen tube can extend into the ovary;
FIG. 3 is a fluorescent micrograph of pollen germination and pollen tube elongation in a style after pollination of the various styles of the present application in example 6; wherein a) is a cylindrical style showing that a large number of pollen normally germinates on the stigma; b) The column is cylindrical, and shows that a large number of pollen tubes grow smoothly in the column; c) The column is a cylindrical style, and shows that a large number of pollen tubes smoothly extend into the ovary; d) The pollen tube is a flat columella, shows that pollen germinates on the stigma, and part of the pollen tube extends into the columella; e) The display part of the pollen tube extends in the flat column; f) The flower column is a flat column, and the pollen distributing pipe is shown to extend into the ovary; g) The column is cracked, the pollen is shown to germinate on the column head, and a small amount of pollen tube extends into the column; h) A split-column style shows a small number of pollen tubes growing in the style; i) The column of the flower is cracked, the pollen tube is shown to be stopped at the junction of the column of the flower and the ovary, and a very small amount of the pollen tube can extend into the ovary;
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be clearly and completely described below with reference to the accompanying drawings of the embodiments of the present invention. The components of embodiments of the present invention generally described and illustrated in the figures herein may be arranged and designed in a wide variety of different configurations.
Thus, the following detailed description of the embodiments of the present invention, presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
Technical means which are not described in detail in the present application and are not intended to solve the technical problems of the present application are provided according to common general knowledge in the art, and various common general knowledge arrangement modes can be implemented.
Examples
Seed resources, reagents and instruments used in the following examples are all obtained from commercial sources unless otherwise specified.
Example 1MS Yunyan 87 Style with various forms, pistil percentage, capsule size, single fruit grain number, thousand grain weight and germination percentage test
Test 1.MS pollination test of Yunyan 87 by different pistil forms
In 2020, seven pistils of different types of MS Yunyan tobacco 87 are pollinated in Xishuangbanna winter breeding base of Nicotiana sempervirens, the pistil forms used in experiments are shown in figure 1, the fruit setting rate, the capsule size (see capsule length and capsule width), the single-fruit number, the thousand-grain weight and the germination rate of offspring of various pistils are counted, the obtained results are shown in the table, and the two divisions of pistil cylinders of sterile lines and the pistil forms of fertile lines are used as comparison treatment.
Table 1 statistics of fructification after pollination of various pistils of MS Yunyan 87
The results show that: pistil morphology was highest in seating rate, capsule width, capsule weight and single grain number for cylinder double split (fig. 1 a), cylinder triple split (fig. 1 b) and cylinder multiple split (fig. 1 c), and the differences between the three pistil morphologies were not significant. The fruit setting rate, capsule width, capsule weight and single fruit grain are repeated several times, namely the flower pillar is flat and long (figure 1 c) and the flower pillar is flat and short (figure 1 e), and the difference between the two pistil forms is not obvious.
The fruit set rate, capsule width, capsule weight and number of individual grains were lowest for boll dehiscence (FIG. 1 f) and boll segregation (FIG. 1 g), and the difference between the two pistil morphologies was not significant.
Before the thousand seed weight and the germination rate are measured in each treatment, the seeds are subjected to air separation and screening, and the difference between the thousand seed weight and the germination rate is not obvious in each treatment.
Therefore, the female characters of the sterile line, which cause the reduction of the seed yield and quality in the MS Yunyan 87 seed reproduction production, are mainly flat and long style, flat and short style, split style and style separation.
Example 2MS Yunyan 87 types of pistil pollination test
Through analysis and combing of early-stage experimental results, the pistils of the sterile line with two cracks in the cylinder, three cracks in the cylinder and multiple cracks in the cylinder show the consistent rule on the pollination fruit setting rate and the single fruit grain number, while the flat length of the style and the flat short length of the style, the cracking of the style and the separation of the style also show the consistent performance on the pollination fruit setting rate and the single fruit grain number respectively.
In 2021, the company, at Xishuangbanna winter breeding base, classified the pistils shown in FIG. 1 by cylinders (FIGS. 1 a-c), flat columns (FIGS. 1 d-e), and split columns (FIGS. 1 f-g), repeated the previous year's experiment, and the results are shown in the following table.
TABLE 2 pollination of different pistil types of MS Yunyan 87
The result shows that the fruit setting rate and the single fruit grain number of the MS Yunyan 87 cylindrical pistil after pollination are the highest, and the MS Yunyan 87 cylindrical pistil is the next, the MS Yunyan pistil is the lowest, and the difference is obvious; the thousand grain weight and the germination rate of each treated seed have no obvious difference.
From this, it is known that the sterile line pistil with cylindrical style is normal pistil, and the pistil with flat or split style is deformed pistil, which can significantly reduce the fruit set rate and the number of single fruit grains.
The distortion rate of the sterile line pistil definition is calculated according to the following formula: pistil distortion rate = (number of deformed pistils/(number of normal pistils + number of deformed pistils)) x100%.
EXAMPLE 3 pollen germination and pollen tube elongation in the flower post after pollination of various types of pistils
Mature pollen of MS Yunyan 87 is collected, a strong MS Yunyan 87 male sterile line plant is selected, and stigma smearing pollination is carried out on cylindrical pistil, oblate pistil and split pistil (defined as example 2) of the sterile line respectively. More than 100 flowers are pollinated in each treatment, about 30 flowers are collected respectively after pollination for 4 and 48 hours, and the petals are removed and then preserved by FAA stationary liquid.
Softening with 1mol/L NaOH for 60min in laboratory, and adding 1g/L water-soluble aniline blue-K 3 PO 4 Stained for more than 1h, mounted on a glass slide and pressed into a sheet and observed by an Olympus BX51 fluorescence microscope, and the results are shown in FIGS. 2 a-i).
The results show that after pollen germinates on the pistil stigma of the cylindrical sterile line, a large number of pollen tubes can smoothly grow in the style and extend into the ovary.
After pollen germinates on the stigmatic gynoecium, only part of the gynoecium can grow in the stigmatic and extend into the ovary.
The pollen only has a small amount of pollen germination on the pistil stigma of the split column type, and only a small amount of pollen tubes grow in the style, most of the pollen tubes can stay at the junction of the style and the ovary, and only a small amount of pollen tubes can extend into the ovary.
Example 4MSK326 different pistil morphological pollination test
In 2020, the MSK326 gynoecium of each type shown in FIG. 1 is pollinated at Xishuangbanna winter breeding base, and the fruit set rate, capsule size, single fruit grain number, thousand grain weight and germination rate of offspring of gynoecium of each type are counted, and the results are shown in the table below. The two divisions of the sterile line pistil cylinder are the same as the shape of the fertile line pistil, and the treatment is a contrast treatment.
Table 3 statistics of fructification after pollination of MSK326 pistils
The results show that after the pistils with different forms of MSK326 are pollinated, the fructification condition is consistent with the expression of MS Yunyan 87, namely the fructification characteristics of fruit setting rate, capsule width, capsule weight and single-fruit number: the two-split cylindrical, three-split cylindrical and multi-split cylindrical flower pole has the advantages that the flat length of the flower pole and the flat short of the flower pole have the advantages that the flower pole is split and separated, the difference between the forms of the three pistils of the two-split cylindrical, three-split cylindrical and multi-split cylindrical flower pole is not obvious, the difference between the forms of the two pistils of the flat length of the flower pole and the flat short of the flower pole is not obvious, and the difference between the forms of the two pistils of the split cylindrical flower pole and the two pistils of the separated cylindrical flower pole is not obvious.
Before the thousand seed weight and the germination rate are measured in each treatment, the seeds are subjected to air separation and screening, and the difference between the thousand seed weight and the germination rate is not obvious in each treatment.
Therefore, the pistil characters of the sterile line, which cause the reduction of the yield and quality of the seeds produced by the MSK326 breeding, are mainly flat and long style, flat and short style, cracked style and separated style.
Example 5 pollination test of various types of pistils in MSK326
Through the analysis and combing of the early-stage experimental results, the sterile line pistil shows consistent cylinder two-crack, cylinder three-crack and cylinder multi-crack on the pollination fruit setting rate and the single fruit grain number, the style is flat long and the style is flat short, the style crack and the style separation are respectively classified into a cylinder, a flat column and a cracked column, and repeated experiments are carried out by a company in the Xishuangbanna winter breeding base in 2021, and the obtained results are shown in the table below.
Table 4 pollination of different pistil types by MSK326
The result shows that the fruit setting rate and the single fruit grain number of the MSK326 cylindrical pistil after pollination are the highest, and the flat-column pistil and the split-column pistil are the lowest, and the difference is obvious; the thousand seed weight and the germination rate of each treated seed have no obvious difference. MSK326 and MS yunyan 87 performed consistently and consistently with previous year results.
From this, the sterile line pistil whose style is cylindrical is defined as normal pistil, and the sterile line pistil whose style is oblate and dehiscent is defined as deformed pistil which can significantly reduce the fruit set rate and the number of single fruit grains, and the deformation rate is calculated as pistil deformation rate = (number of deformed pistil/(number of normal pistil + number of deformed pistil)) x100%.
Example 6 pollen germination and pollen tube elongation in the style after pollination of various types of pistils
Collecting MSK326 mature pollen, selecting strong MSK326 male sterile line plants, and respectively carrying out stigma smearing pollination on cylindrical pistils, oblate pistils and schizont pistils of the sterile line. More than 100 flowers are pollinated in each treatment, more than 30 flowers are collected after pollination for 4 hours and 48 hours, and the flowers are preserved by FAA stationary liquid after petals are removed.
The sample was taken back to the laboratory and softened with 1mol/L NaOH for 60min, stained with 1g/L water-soluble aniline blue-K3 PO4 for more than 1h, placed on a glass slide and pressed into a sheet, and observed with an Olympus BX51 fluorescence microscope. The result shows that after pollen germinates on the pistil stigma of the cylindrical sterile line, a large number of pollen tubes can smoothly grow in the style and extend into an ovary; after pollen germinates on the flat-pillar type female pistil stigma, only part of the female pistil can grow in the style and extend into an ovary; and only a small amount of pollen germinates on the split-pillar type female stamen head and only a small amount of pollen tube grows in the stylobate, and most of the pollen tube stays at the junction of the stylobate and the ovary, and only a very small amount of pollen tube can extend into the ovary. MSK326 performed in agreement with MS Yunyan 87, and the results are shown in FIGS. 3 a-i).
Example 7 determination of Effect of applying exogenous Material on improving pistil abnormality
After defining the calculation method of the type of the gynoecium malformed gynoecium and the gynoecium teratogenicity rate of the sua cytoplasmic sterile line, 2021, the company tried to apply different treatments on MS yunyan 87 to investigate how to improve gynoecium malformation. Preparation of reagents used for each treatment: 10umol/LMeJA (prepared by dissolving 2.243mg of analytically pure MeJA in 1L of water), 1/1000 borax (prepared by dissolving 1g of borax in 1L of water), and 1g/L potassium dihydrogen phosphate (prepared by dissolving 1g of analytically pure potassium dihydrogen phosphate in 1L of water).
All treatments were: removing all large buds in the full-bloom stage of the tobacco plant, and only leaving small buds just differentiated;
the whole leaf surface is evenly sprayed, 10ml of the fertilizer is applied to each plant, 3 treatments are set for each treatment, and after the first flowering, all large buds and the bloomed flowers are collected for counting the abnormal rate. After the second flowering, all large flower buds and the opened flowers were collected as well for statistical aberration rates, and the results of the effects of different treatments on the rosette deformity are shown in the table below.
TABLE 5 Effect of different treatments on pistil distortion Rate of MS Yunyan 87
As can be seen from the table above, the first flowering and fruiting shows that the shape rate of the monopotassium phosphate leaf surface spraying treatment with the concentration of 1g/L is 17.14% lower than that of the control group, and the difference is obvious; and the pistil aberration rates of 10umol/LMeJA and 1/1000 concentration borax treatment are not obviously different from the control.
The second batch of flowering results shows that the stamen aberration rates of the three treatments are not obviously different from the control; the difference of the flowering time of the two batches is 10 days, which shows that the influence of 1g/L monopotassium phosphate on the pistil aberration rate is less than 10 days.
EXAMPLE 8 determination of optimum Potassium dihydrogen phosphate concentration
On the basis of example 7, potassium dihydrogen phosphate solutions with concentrations of 0.1g/L, 1g/L and 5g/L are respectively set, all large buds are removed in the full-bloom period of the tobacco plant, only small buds just differentiated are left, the whole leaf surface is uniformly sprayed, 10ml of the solution is applied to each plant, 3 treatments are set for each treatment, and after blooming, all large buds and the opened flowers are collected, and the statistical aberration rate is shown in the following table.
TABLE 6 influence of different concentrations of potassium dihydrogen phosphate on pistil aberration rate of MS Yunyan 87
| Treatment of | Pistil distortion ratio (%) |
| 0.1g/L potassium dihydrogen phosphate | 55.93b |
| 1g/L potassium dihydrogen phosphate | 47.40c |
| 5g/L Potassium dihydrogen phosphate | 48.68c |
| Contrast (clean water) | 65.34a |
The results show that different concentrations of the monopotassium phosphate have obvious effects on the improvement of the deformity of the MS Yunyan 87, and the effect is improved firstly and then is unchanged along with the increase of the concentration of the monopotassium phosphate. As can be seen, the concentration of potassium dihydrogen phosphate applied was the optimum concentration at 1g/L.
Example 9
In 2021, the company found that the gynoecium aberration rate of continuous cropping in the whole year is 31.11% higher than that of the gynoecium in the frequent rotation in conventional management of breeding field. After the quick-acting potassium and the total potassium in the two pieces of field soil are tested, the content of the quick-acting potassium in the field soil which is continuously planted all year round is 17.03 percent higher than that of the field which is frequently planted, and the content of the total potassium is 16.05 percent higher than that of the field which is frequently planted under the same fertilization and management level. Continuous cropping is presumed to affect the absorption of potassium in the soil by tobacco plants, so that the potassium remaining in the soil is higher than that in a crop rotation field, and the reason that the pistil aberration rate of continuous cropping fields is higher is related to the low potassium absorption of the tobacco plants.
The tobacco plants in the two fields are sprayed with 1g/L potassium dihydrogen phosphate solution at the same time, and the pistil aberration rate of the rotation field is found to be reduced by 17.93 percent, and the pistil aberration rate of the continuous cropping field is found to be reduced by 23.33 percent. The spraying of the monopotassium phosphate shows that the pistil aberration rate can be obviously reduced, and the reduction range is larger than that of the crop rotation field. The results obtained are shown in the following table:
TABLE 7 survey of potassium content and pistil aberration rate in tobacco planting soil of different fields
From the above, the solution to the above problems can be achieved by spraying a potash fertilizer on the liquid surface of a tobacco plant planted in the tobacco continuous cropping soil, and the specific spraying method can be applied according to a common method in the field.
Example 10
In 2022, the company expanded the reagent screening range to further explore the influence of exogenous substances on gynoecium.
All treatments were: removing all large flower buds in the full-bloom stage of the tobacco plant, and only leaving small flower buds which are just differentiated; uniformly spraying the corresponding solution in each treatment in the table 8 on the whole leaf surface, wherein the solvent used in the solution in each treatment is water, and the solution in each treatment is prepared:
dissolving 22.43mg of analytically pure methyl jasmonate in 1L of water; 20mg of analytically pure potassium indolebutyrate was dissolved in 1L of water; 1L of water was dissolved with 30mg of analytically pure proline; 2.5g of borax is dissolved in 1L of water; 1gMS culture medium is dissolved in 1L water; 0.555g of analytically pure calcium chloride is dissolved in 1L of water; 20mg of analytically pure melatonin was dissolved in 1L of water; 1g of analytically pure potassium dihydrogen phosphate was dissolved in 1L of water; 25mg of gibberellin was dissolved in 1L of water.
The spraying method of each treatment comprises the following steps: each plant was applied with 10ml of each treatment solution, each treatment was set to 3 replicates, statistical aberration rates were collected for all large flower buds and bloomed flowers after the first flowering, statistical aberration rates were also collected for all large flower buds and bloomed flowers after the second flowering, and the effects of different treatments on stigma deformity are shown in the table below.
TABLE 8 Effect of different treatments on MSK326 pistil malformation
| Treatment of | First pistil distortion (%) | Second batch pistil distortion ratio (%) |
| Contrast (clean water) | 64.46a | 66.19a |
| 100umol/L jasmonic acid methyl ester | 65.42a | 64.74a |
| 20mg/L Potassium indolebutyrate | 64.29a | 62.84a |
| 30mg/L proline | 64.04a | 65.42a |
| 2.5/1000 borax | 61.58ab | 63.37a |
| 1/1000MS culture medium | 61.46ab | 63.10a |
| 5mmol/L calcium chloride | 61.14ab | 62.97a |
| 20mg/L melatonin | 58.29ab | 61.64a |
| 1g/L potassium dihydrogen phosphate | 47.05b | 62.20a |
| 25mg/L gibberellin | 43.81b | 61.59a |
As can be seen from the above table, the first flowering results show that 2.5/1000 borax, 1/1000MS culture medium, 5mmol/L calcium chloride, 20mg/L melatonin, 1g/L monopotassium phosphate and 25mg/L gibberellin all have an improvement effect on MSK326 gynoecium malformation, wherein 1g/L monopotassium phosphate and 25mg/L gibberellin can significantly reduce the MSK326 gynoecium malformation rate by 17.41% and 20.65% respectively compared with the control.
However, further studies have found that gibberellin treatment has a great influence on MSK326 flowering, the flowers become long and narrow, the flower color is deepened, the number of flowering is severely reduced, and the gibberellin treatment is not suitable for application in production.
The second batch of flowers and fruits have insignificant treatment difference, and the time interval between the two batches of flowers is 10 days. Therefore, the improvement effect of the sua sterile cytoplasmic pistil deformity is still 1g/L of the monopotassium phosphate, and the action time is not more than 10 days.
Although the present invention has been described in detail with reference to the foregoing embodiments, it will be apparent to those skilled in the art that various changes in the embodiments and/or modifications of the invention can be made, and equivalents and modifications of some features of the invention can be made without departing from the spirit and scope of the invention.
Claims (9)
1. A method for reducing the pistil aberration rate and breeding and planting a cytoplasmic male sterile line of tobacco sua is characterized by comprising the following steps: spraying KH on the leaf surface of the whole tobacco plant before the bud of the tobacco plant 2 PO 4 Spraying solution with at least 10 mKH per tobacco 2 PO 4 A solution;
sprayingKH 2 PO 4 The concentration of the solution is more than or equal to 0.1g/L.
2. The method for decreasing pistil aberration rate and breeding and planting of cytoplasmic male sterile line in tobacco sua according to claim 1, wherein KH is sprayed once 2 PO 4 The interval time of the solution is 5-10 days until the tobacco seed reproduction and pollination period is finished.
3. The method for decreasing the pistil aberration rate and breeding and planting of a tobacco sua cytoplasmic male sterile line of claim 1, wherein the pistil aberration rate of the sua sterile line is decreased by more than 15% by the method.
4. The method for decreasing the pistil aberration rate and breeding and planting of cytoplasmic male sterile line of tobacco sua as claimed in claim 3, wherein the aberration rate is calculated by the following formula:
pistil distortion rate = (number of malformed pistils/(number of normal pistils + number of malformed pistils)) x100%, where normal pistils refer to: the flower column is circular, and the column head is divided into two parts; or the flower column is round, and the column head is three-split; or pistil with round style and cracked head;
malformed pistils refer to: the flower pillar is flat and short; or the flower column is flat and long; or the flower column is cracked; or pistils separated from the style.
5. The method for decreasing pistil aberration rate and breeding and planting of tobacco sua cytoplasmic male sterile line according to claim 3, wherein the sterile cytoplasm of the male sterile line tobacco variety is derived from sweet tobacco (N.
6. The method for decreasing pistil aberration rate and breeding and planting of tobacco sua cytoplasmic male sterile line according to claim 3, wherein the tobacco sua cytoplasmic male sterile line is MS Yunyan 87, MSK326.
7. The method for decreasing pistil aberration rate and breeding and planting of cytoplasmic male sterile line of tobacco sua as claimed in claim 3, wherein KH is a natural number of plants 2 PO 4 The concentration of the solution is 0.1-5 g/L.
8. The method for decreasing pistil aberration rate and breeding and planting of cytoplasmic male sterile line in tobacco sua as claimed in claim 3, wherein KH is defined as 2 PO 4 The concentration of the solution was 1g/L.
9. The method for decreasing pistil aberration rate and breeding and planting of cytoplasmic male sterile line of tobacco sua as claimed in claim 3, wherein said tobacco plant is planted in soil for continuous cropping of tobacco.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4709710A (en) * | 1978-09-05 | 1987-12-01 | Fabriques De Tabac Reunies S.A. | Process for improving tobacco |
| WO2004082366A2 (en) * | 2003-03-19 | 2004-09-30 | Regents Of The University Of Minnesota | Methods to confer enhanced floral properties to plants |
| CN102106256A (en) * | 2011-01-21 | 2011-06-29 | 贵州大学 | Hybrid seed production method for tobacco |
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2022
- 2022-10-31 CN CN202211347175.0A patent/CN115918516B/en active Active
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4709710A (en) * | 1978-09-05 | 1987-12-01 | Fabriques De Tabac Reunies S.A. | Process for improving tobacco |
| WO2004082366A2 (en) * | 2003-03-19 | 2004-09-30 | Regents Of The University Of Minnesota | Methods to confer enhanced floral properties to plants |
| CN102106256A (en) * | 2011-01-21 | 2011-06-29 | 贵州大学 | Hybrid seed production method for tobacco |
Non-Patent Citations (1)
| Title |
|---|
| 任雅君等: "喷施KH_2PO_4对盆栽花烟草开花的影响", 安徽农业科学, vol. 38, no. 10, pages 5084 - 5085 * |
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