CN114568066B - Method for screening cotton emergent seedling stage cold-resistant varieties and application thereof - Google Patents
Method for screening cotton emergent seedling stage cold-resistant varieties and application thereof Download PDFInfo
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- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
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- A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
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- A—HUMAN NECESSITIES
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Abstract
The invention relates to the technical field of cotton variety screening, in particular to a method for screening a cotton germination emergence period cold-resistant variety and application thereof. The screening method comprises the steps of continuously performing low-temperature treatment on cotton varieties to be selected in a germination period and a bud period; and carrying out sudden cooling treatment on cotton varieties to be selected in the bud stage. According to the cotton seed germination period, the growth characteristics of the germination period and the weather characteristics of the cold in the spring, the method firstly introduces the treatment of combining sudden cooling and continuous low temperature consistent with the field low temperature generation mode to carry out cotton variety screening, and can obviously improve the screening accuracy and the screening efficiency; meanwhile, the invention determines the performance index which has obvious influence on the cold resistance of germination emergence period. The method disclosed by the invention is high in accuracy, relatively few in identification index, relatively simple to operate and higher in screening efficiency. The screening method can ensure that the cotton seeds still maintain good growth situation after being subjected to 'cold in the back spring', and has important production significance for one-seeding whole seedlings.
Description
Technical Field
The invention belongs to the technical field of cotton variety screening, and particularly relates to a method for screening a cotton germination emergence period cold-resistant variety and application thereof.
Background
Cotton is native to subtropical zones and is warm and cold-resistant. However, in the main cotton production area of China, cold and wet air flow activities in the northwest direction are frequent every 4 to 5 months, so that the air temperature is greatly fluctuated, and the final frost period is unstable, namely commonly called' cold in spring. The low temperature of more than or equal to 0 ℃ and less than or equal to 15 ℃ is easy to form, and the rotted seed and rotted bud are caused in the emergence period, so that the emergence rate is low; the cotton plant grows slowly and is frozen and killed in the cotyledon period after emergence of seedlings, and the low temperature in early spring has become one of the main environmental factors influencing the production of Xinjiang cotton. It is therefore necessary to screen out the cold-tolerant varieties of cotton to ensure cotton yield.
In the past, it has been empirically believed that the seedling stage is a critical stage of cotton cold damage, and thus, the screening of existing cotton cold-tolerant varieties is performed for the cotton seedling stage (i.e., cotyledon stage), such as CN110073974a. However, the inventors of the present application found that the period of emergence hazard is long and strong compared to the low temperature hazard after emergence. Sowing is carried out when the temperature of the soil layer under the film reaches 12 ℃ steadily, and cotton seeds are always in a low-temperature stress environment until the seedlings emerge. Statistical analysis of meteorological data in the cotton district 2009-2020 in northern Xinjiang finds that the average temperature of the days is 10-12 ℃ low-temperature hazard in 4 months 5-12 days, the minimum temperature of the days 18-20 months suddenly drops from 8 ℃ to 4 ℃, rebroadcasting is needed when the hazard is serious, the yield can be reduced by 20% -60%, and the low-temperature cold hazard in the seedling stage seriously affects the cotton production. In addition, through early work, it is found that cotton varieties with higher cold resistance in the germination seedling stage, which are different in the seedling stage due to different development stages, do not necessarily have higher cold resistance, so that an identification method suitable for the germination seedling stage of cotton is necessary to be formulated.
Disclosure of Invention
In order to solve the technical problems, the invention particularly provides a novel screening method of cotton cold-resistant varieties. The cotton seeds obtained by screening through the method still maintain good growth situation after being subjected to 'cold in the back spring', and have important production significance for one-seeding whole seedlings.
The invention relates to a screening method of cotton cold-resistant varieties, which comprises the following steps: carrying out low-temperature stress treatment on cotton varieties to be selected in the germination period; performing index collection, data processing and analysis on the processed cotton varieties; determining the cold resistance level of the cotton variety to be selected according to the data processing and analysis results; wherein the germination period comprises: germination and bud phase; the low temperature stress treatment comprises: and continuously performing low-temperature treatment on the cotton varieties to be selected in the germination period and the bud period, and performing sudden cooling treatment on the cotton varieties to be selected in the bud period.
Through the intensive research on the growth characteristics of cotton and the temperature characteristics of the cold in the spring, the invention provides a screening mode of suddenly cooling treatment for the first time, and combines the screening mode with continuous low-temperature treatment, so that the screening accuracy and the screening efficiency can be obviously improved, the identification of cold-resistant varieties in the germination period can be developed, the negative effects caused by the cold-resistant varieties can be counteracted by making corresponding strategies, and the cotton seeds can still maintain good growth situation even suffering from the cold in the spring, thereby having important production significance for one-plant whole seedlings.
Meanwhile, in order to more efficiently and accurately screen out the cold-resistant varieties, the invention optimizes the matching relation between the growing period of the cotton varieties and the low-temperature treatment mode, and proposes continuous low-temperature treatment in the germination period and the bud period and abrupt cooling treatment in the bud period. Through reasonable configuration of cotton growing period and low-temperature treatment mode, operation can be further simplified while screening accuracy is ensured, and screening efficiency is improved.
Preferably, the conditions of the continuous low temperature treatment are: controlling the temperature of day culture at 15-16deg.C for 12 hr, and night culture at 3-4deg.C for 12 hr; the treatment time of the continuous low-temperature treatment is 12-14 days. Research shows that compared with the constant temperature treatment mode adopted by the conventional screening method, the continuous low-temperature treatment in the day-night temperature difference mode can be used for more fully knowing the cold resistance of the cotton varieties to be selected, so that the accuracy of the screening result is further improved.
Preferably, the conditions of the sudden cooling treatment are as follows: the culture temperature is controlled between 3-4deg.C, and the treatment time is 3-4 days. Research shows that the cold resistance of the cotton varieties to be selected can be more fully known through short-time low-temperature treatment, and the accuracy of screening results is further improved.
It is known in the art that the more screening indexes, the more accurate the screening result is; however, excessive indexes firstly cause large manual operation errors, and secondly are complex in operation, time-consuming and labor-consuming. Therefore, it is a technical difficulty of the present application how to obtain more accurate screening results with fewer indexes and a simpler and more convenient method.
In order to solve the problem, the method determines the following proper indexes from a plurality of phenotype indexes and physiological indexes of cotton varieties through a large number of test analysis, so that the cold resistance of the cotton varieties to be selected in the germination period and the bud period can be comprehensively reflected, the acquisition operation is simplified, the manual error is reduced, and the screening accuracy is ensured. The specific indexes are as follows:
for sustained low temperature treatment, the index is 7 in total, and the index is respectively: germination rate (GR-CC), germination index (GI-CC), emergence rate (ER-CC), root length (RL-CC), lower embryo axial length (HL-CC), total length (TL-CC), dry weight (DW-CC);
for the sudden cooling treatment, the indexes are 5, and the indexes are respectively as follows: emergence rate (ER-SC), root length (RL-SC), axis length of embryo (HL-SC), total length (TL-SC), dry weight (DW-SC).
The invention adopts a sand culture method culture mode, and particularly comprises two culture modes of a culture dish sand culture method and a paper bowl sand culture method:
collecting germination rate and germination index of continuous low-temperature treatment by adopting a culture dish sand culture method;
and collecting the number of seedlings, root length, axis length of the embryo, total length and dry weight of the continuous low-temperature treatment and sudden cooling treatment by adopting a paper bowl sand culture method.
Specifically, for the collection of germination rate and germination index for continuous low temperature treatment, specifically: and (3) adopting a culture dish sand culture method, placing the sown cotton varieties to be selected in a continuous low-temperature treatment environment for treatment, recording the germination number every 2 days from germination, and stopping recording after the germination number is stable and unchanged.
The collection of the number of seedlings, root length, axis length of embryo, total length and dry weight of continuous low-temperature treatment is as follows: and (3) adopting a paper pot sand culture method, placing the sown cotton varieties to be selected in a continuous low-temperature treatment environment for treatment, transferring the cotton varieties to 25-28 ℃ for at least 3 days after the continuous low-temperature treatment is finished, and recording.
The collection of the emergence number, root length, embryo axial length, total length and dry weight of the sudden cooling treatment is specifically as follows: culturing in paper bowl sand culture at 25/28deg.C under light/dark culture for 12/12 hr to bud stage, transferring cotton variety to be selected in bud stage into suddenly cooling treatment environment, transferring to normal temperature environment, recovering growth, and recording.
Further, in order to eliminate the difference between the basic traits of different germplasm, the invention processes the data of the acquired trait indexes, and comprises the following steps:
(1) Calculating the relative value of the index (namely the cold resistance coefficient CC);
cold tolerance coefficient CC (Cold tolerance coefficient) = treatment measure/control measure;
(2) Calculating a membership function value U;
membership function value U ij ＝(C ij -C min )/(C max -C min )；
Wherein:
C ij : j-th common factor score of i materials, C min Representing the minimum value of all germplasm j common factor score values;
C max representing the maximum value of the scoring values of all germplasm j common factors;
C ij is calculated by EXCLE from the cold resistance coefficient CC.
(3) Calculating a comprehensive evaluation value D of cold resistance;
wherein:
di represents the comprehensive evaluation value of the ith material;
U ij membership function value of the jth common factor for the ith material;
W j the importance degree of the jth common factor in all common factors is represented, namely a weight coefficient;
weight coefficient
Pj is the contribution of the jth common factor.
(4) Based on the D values, clustering analysis was performed using the euclidean distance and weighted paired arithmetic averaging method (weighted pair group method average, WPGMA). And determining the cold resistance level of the cotton variety according to the analysis result.
The invention also provides a cotton breeding method adopting the screening method, and the breeding efficiency and accuracy can be effectively improved through the method.
The screening method and the breeding method are suitable for different cotton producing areas, and are particularly suitable for the cotton producing areas in Xinjiang.
The beneficial effects obtained by the invention are as follows:
(1) The invention provides a method for screening cold-resistant varieties aiming at cotton germination sprouting period (including sprouting period and sprouting period) through analysis and research on field low-temperature cold injury action forms.
(2) The screening method disclosed by the invention is high in accuracy, relatively few in identification index, relatively simple to operate and higher in screening efficiency.
Detailed Description
The following examples are illustrative of the invention and are not intended to limit the scope of the invention.
Example 1
The embodiment provides a screening method of cotton cold-resistant varieties, which comprises the following steps:
s1, cotton varieties to be selected are shown in the table:
TABLE 1
| Numbering device | Variety name | Numbering device | Variety name | Numbering device | Variety name | Numbering device | Variety name |
| 1 | Dai-shaped cotton 20 | 26 | Xinlunzao No. 4 | 51 | Dai character cotton No. 14 | 76 | U.S. 1 |
| 2 | Guangong No. 1 | 27 | Xinlunzao No. 6 | 52 | Dai character cotton No. 15 | 77 | Gilgis |
| 3 | Handan 2490 | 28 | Xinlunzao No. 8 | 53 | Cotton No. 3 | 78 | Cultivation 6614 |
| 4 | Black mountain cotton No. 1 | 29 | Xinlunzao No. 9 | 54 | Guoxin cotton No. 11 | 79 | Reclamation 0074 |
| 5 | Brocade cotton No. 3 | 30 | Relaxation 1335 | 55 | Ji 4025 | 80 | Chuan 01 |
| 6 | Jincotton No. 10 | 31 | Early relaxation 8E13 | 56 | Jin cotton 26 | 81 | Xinlunzao No. 2 |
| 7 | Jin cotton 21 | 32 | Early fortune N177 | 57 | Liaocotton No. 23 | 82 | Xinlunzao No. 10 |
| 8 | Jin cotton 23 | 33 | Early fortune N95 | 58 | Liaocotton No. 27 | 83 | Xinlunzao No. 12 |
| 9 | Jinyan No. 5 | 34 | Middle 716 | 59 | Lu cotton grinding 21 | 84 | Xinlunzao No. 13 |
| 10 | Liaocotton No. 10 | 35 | Middle cotton station 14 | 60 | Lu cotton grinding 28 | 85 | Xinlunzao No. 15 |
| 11 | Liaocotton No. 5 | 36 | Middle cotton house 16 | 61 | Lu cotton grinding 36 | 86 | Xinlunzao No. 16 |
| 12 | Liaocotton No. 6 | 37 | Middle cotton house 20 | 62 | Kernel and 39 # | 87 | New Liu Zao No. 17 |
| 13 | Liaocotton No. 7 | 38 | Middle cotton house 24 | 63 | Shaan 79 | 88 | Xinlunzao No. 18 |
| 14 | Liaocotton No. 9 | 39 | Middle cotton house 27 | 64 | Style cotton 2B | 89 | New land early No. 19 |
| 15 | Liaoyang pup joint | 40 | Middle cotton house 30 | 65 | New cotton 33B | 90 | Xinlunzao No. 20 |
| 16 | Robust 890 | 41 | Middle cotton house 37 | 66 | Honeysuckle tree | 91 | Xinlunzao 21 |
| 17 | Agricultural reclamation No. 5 | 42 | CG3020-1 | 67 | Middle 17 | 92 | Xinlunzao 22 |
| 18 | Shaan 70 | 43 | CG3020-3 | 68 | 19 in (F) | 93 | Xinlunzao No. 23 |
| 19 | Shizao No. 1 | 44 | FM1735 | 69 | 662 in the middle | 94 | Sichuan 239-1 |
| 20 | Shizao No. 2 | 45 | H109 | 70 | Middle 679 | 95 | Yu cotton No. 5 |
| 21 | Early summer No. 1 | 46 | PHy-7 | 71 | Middle cotton house No. 12 | 96 | Fortune 1729 |
| 22 | Xinlunzao 11 | 47 | STS458 | 72 | Middle cotton institute No. 7 | 97 | Zhejiang cotton No. 11 |
| 23 | Xinlunzao 36 | 48 | TM-1 | 73 | Middle cotton planting No. 8 | 98 | T-spread |
| 24 | Xinlunzao 42 | 49 | Orthosiphon 4406 | 74 | 920346 of Shaan | 99 | Bo 425 |
| 25 | Xinlunzao 45 | 50 | Bai Gong 985 | 75 | Limit III9704 | 100 | Xinlunzhong No. 60 |
TABLE 2
S2, carrying out low-temperature stress treatment on cotton varieties to be selected, wherein the method specifically comprises the following steps:
(1) Continuously performing low-temperature treatment on germination period and bud period of cotton varieties to be selected;
the germination and sprouting conditions of 191 parts of materials under the condition of continuous low temperature (16/4 ℃) of day and night change are researched by using a sand culture method, and related indexes of germination and sprouting are needed to be measured by adopting two culture modes:
the culture dish sand culture method is used for measuring germination related indexes (germination rate and germination index), the germination number is counted after sowing and placed at a low temperature of 16/4 ℃ for 5 days, and then counting is carried out once every 2 days until the germination number is stable;
the number of seedlings, root length, axis length of embryo, total length and dry weight are measured by paper pot sand culture, and the culture and design method are the same as those of culture dish sand culture.
Culturing at a proper temperature of 28/25 ℃ after low-temperature germination for 15 days, counting the emergence rate after 3 days, and randomly selecting 10 representative seedlings to measure the seedling length and the seedling dry weight; the data are recorded in the following table.
The influence of continuous low-temperature treatment on cotton seed germination is examined, and the calculation formula is as follows:
obtaining the relative value of each index according to the calculation method of the cold resistance coefficient CC;
RGI (relative germination index) =gr_c/gr_n
GI (germination index) = Σ (Gt/Dt) (Gt is the number of germination on day t, dt is the corresponding number of germination days), gi_c represents the treatment group germination index, and gi_n represents the control group germination index.
The relative values of the indexes including germination rate, emergence number, root length, axis length of the embryo, total length and dry weight are obtained according to the calculation method of the cold resistance coefficient.
TABLE 3 continuous cooling record results (1)
TABLE 4 continuous cooling record result (2)
(2) Carrying out sudden cooling treatment on the bud period of the cotton variety to be selected;
the invention discovers that 20-30 hours after germination is sensitive to cold injury and has great influence on growth and development, so that the influence of low temperature on hypocotyl elongation is explored in the germination period.
The method comprises the following specific steps: the material seeds are germinated for 48h at a proper temperature (28/25 ℃) to a bud phase, the material seeds are shifted to 4 ℃, the hypocotyl length is measured after cold treatment for 4 days, then the material seeds are transferred to a normal temperature (28/25 ℃) to carry out recovery growth, and indexes such as the number of seedlings, the length of the seedlings and the like are counted after the recovery growth for 4 days. The data are recorded in the following table.
TABLE 5 abrupt cooling recording results
S3, data processing and analysis
In order to eliminate the difference between different germplasm basic characters, the invention carries out data processing on all character indexes acquired by the acquisition, and comprises the following steps:
(1) Calculating the relative value of the index (namely the cold resistance coefficient CC);
cold tolerance coefficient CC (Cold tolerance coefficient) = treatment measure/control measure;
(2) Calculating a membership function value U;
membership function value U ij ＝(C ij -C min )/(C max -C min )；
Wherein:
C ij : j-th common factor score of i materials, C min Representing the minimum value of all germplasm j common factor score values;
C max representing the maximum value of the scoring values of all germplasm j common factors;
C ij is calculated from the cold resistance coefficient CC by SPSS.
(3) Calculating a comprehensive evaluation value D of cold resistance;
wherein:
di represents the comprehensive evaluation value of the ith material;
U ij membership function value of the jth common factor for the ith material;
W j the importance degree of the jth common factor in all common factors is represented, namely a weight coefficient;
weight coefficient
Pj is the contribution of the jth common factor.
(4) According to the D value, adopting a Euclidean distance and weighting pairing arithmetic average method (weighted pair group method average, WPGMA) to perform cluster analysis;
s4, determining the cold resistance level of the cotton variety to be tested according to the analysis result.
In order to verify the feasibility of the screening method, the invention establishes the optimal regression equation by taking the cold resistance comprehensive evaluation value D as a dependent variable and the relative value CC of each single index as an independent variable:
regression equation for continuous cooling treatment:
TABLE 6
| Multiple stepwise regression equation | R 2 | F value | P |
| Y＝0.798*GR | 0.635 | 346.160 | ≤0.001 |
| Y＝0.688*GR+0.509*TL-CC5 | 0.883 | 752.380 | ≤0.001 |
| Y＝0.663*GR+0.459*TL-CC5+0.282*DW-CC5 | 0.959 | 1583.776 | ≤0.001 |
Multiple regression equation for sudden cooling treatment:
TABLE 7
| Multiple stepwise regression equation | R 2 | F value | P |
| Y＝0.860*TL-SC6 | 0.739 | 563.656 | ≤0.001 |
| Y＝0.882*TL-SC6+0.329*ER-SC6 | 0.846 | 547.932 | ≤0.001 |
| Y＝0.707*TL-SC6+0.282*ER-SC6+0.320*HL-SC6 | 0.917 | 739.578 | ≤0.001 |
The result shows that the model fitting degree is good, and the regression equation has good interpretation capability; the acquisition index determined by the invention has obvious influence on the cold resistance of the sprouting seedling stage, and the model can be used for evaluating the cold resistance of the sprouting seedling stage.
While the invention has been described in detail in the foregoing general description and with reference to specific embodiments thereof, it will be apparent to one skilled in the art that modifications and improvements can be made thereto. Accordingly, such modifications or improvements may be made without departing from the spirit of the invention and are intended to be within the scope of the invention as claimed.
Claims (6)
1. A method for screening a cotton cold-resistant variety, comprising the steps of:
carrying out low-temperature stress treatment on cotton varieties to be selected in the germination period;
performing index collection and data processing stage analysis on the processed cotton varieties;
determining the cold resistance level of the cotton variety to be selected according to the data processing and analysis results;
wherein the germination period comprises: germination and bud phase;
the low temperature stress treatment comprises: continuously performing low-temperature treatment on cotton varieties to be selected in germination and bud stages, and performing sudden cooling treatment on the cotton varieties to be selected in bud stages;
the conditions of the continuous low-temperature treatment are as follows: controlling the temperature of day culture at 15-16deg.C for 12 hr, and night culture at 3-4deg.C for 12 hr; the treatment time of the continuous low-temperature treatment is 12-14 days;
the conditions of the sudden cooling treatment are as follows: controlling the culture temperature between 3-4deg.C, and treating for 3-4 days;
for sustained low temperature treatment, the indicators are: germination rate, germination index, emergence rate, root length, axis length of embryo, total length, dry weight;
for the sudden cooling treatment, the indexes are as follows: emergence rate, root length, axis length of embryo, total length, dry weight.
2. The method for screening cold-resistant cotton varieties according to claim 1, wherein the continuous low-temperature treatment adopts a sand culture method;
the sand culture method comprises a culture dish sand culture method and a paper bowl sand culture method;
collecting germination rate and germination index of continuous low-temperature treatment by adopting a culture dish sand culture method;
and collecting the number of seedlings, root length, axis length of the embryo, total length and dry weight of the continuous low-temperature treatment and sudden cooling treatment by adopting a paper bowl sand culture method.
3. The method for screening cotton varieties with cold resistance according to claim 2, wherein the collection of germination rate and germination index for continuous low temperature treatment is specifically: and (3) adopting a culture dish sand culture method, placing the sown cotton varieties to be selected in a continuous low-temperature treatment environment for treatment, recording the germination number every 2 days from germination, and stopping recording after the germination number is stable and unchanged.
4. The method for screening cotton varieties with cold resistance according to claim 2, wherein the collection of the number of seedlings, root length, axis length of embryo, total length and dry weight for continuous low temperature treatment is specifically: adopting a paper bowl sand culture method, placing the sown cotton varieties to be selected in a continuous low-temperature treatment environment for treatment, transferring the cotton varieties to 25-28 ℃ for at least 3 days after the continuous low-temperature treatment is finished, and recording;
the collection of the emergence number, root length, embryo axial length, total length and dry weight of the sudden cooling treatment is specifically as follows: culturing for 2d to bud stage at 25/28deg.C under light/dark culture for 12/12h by paper bowl sand culture, transferring cotton variety to be selected in bud stage into suddenly cooling treatment environment, transferring to normal temperature environment, recovering growth, and recording.
5. The method for screening cotton cold-tolerant varieties according to any one of claims 1 to 4, characterized in that the data processing and analysis comprises:
(1) Calculating the relative value of the index;
cold tolerance CC = treatment measurement/control measurement;
(2) Calculating a membership function value U;
membership function value U ij ＝(C ij -C min )/(C max -C min )；
Wherein:
C ij : j-th common factor score of i materials, C min Representing the minimum value of all germplasm j common factor score values;
C max representing the maximum value of the scoring values of all germplasm j common factors;
C ij is calculated by the cold resistance coefficient CC by EXCLE;
(3) Calculating a comprehensive evaluation value D of cold resistance;
wherein:
di represents the comprehensive evaluation value of the ith material;
U ij membership function value of the jth common factor for the ith material;
W j the importance degree of the jth common factor in all common factors is represented, namely a weight coefficient;
weight coefficient
Pj is the contribution rate of the jth common factor;
(4) And (3) carrying out cluster analysis by adopting a Euclidean distance and weighted pairing arithmetic average method according to the D value.
6. A method for breeding a cotton cold-tolerant variety, characterized in that the screening method according to any one of claims 1 to 5 is used.
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