CN109997446B - Method for improving deep sowing resistance of corn seeds - Google Patents

Method for improving deep sowing resistance of corn seeds Download PDF

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CN109997446B
CN109997446B CN201910296554.3A CN201910296554A CN109997446B CN 109997446 B CN109997446 B CN 109997446B CN 201910296554 A CN201910296554 A CN 201910296554A CN 109997446 B CN109997446 B CN 109997446B
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CN109997446A (en
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赵小强
彭云玲
陆晏天
白明兴
徐明霞
姬祥卓
王芳
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Gansu Agricultural University
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01C—PLANTING; SOWING; FERTILISING
    • A01C1/00—Apparatus, or methods of use thereof, for testing or treating seed, roots, or the like, prior to sowing or planting
    • A01C1/08—Immunising seed

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Abstract

A method for improving the deep sowing resistance of corn seeds comprises the following steps: (1) prepared into 0, 0.5, 1.0, 1.5 and 2.0 mg.L‑1And 24-Epibrassinolide (EBR). (2) Uniformly mixing sterilized vermiculite and corresponding EBR solution according to the proportion of 5g:1mL, then respectively filling the mixture into a seed deep-sowing test device at a certain height, sowing 30 seeds soaked by the corresponding EBR solution on the vermiculite, and then respectively covering the seeds with 3 cm, 15 cm and 20 cm of corresponding vermiculite. After sowing, pouring 40 mL of corresponding EBR solution quantitatively every 2 d, and measuring 18 related characters of deep sowing resistance after germinating in a phytotron for 10 d. (3) Newly defining an exogenous EBR deep-sowing-resistant regulation index with a single character, adopting a membership function method, comprehensively evaluating the regulation effect of the exogenous EBR on the maize seed emergence under the deep-sowing stress, laying a theoretical foundation for improving the deep-sowing-resistant characteristic of the maize seed, providing a technical support for resisting drought for the maize planted to the seedling form building stage, and having wide application potential.

Description

Method for improving deep sowing resistance of corn seeds
Technical Field
The invention belongs to the field of agricultural technology application, and particularly relates to a method for improving deep-sowing seedling emergence of corn seeds under the deep-sowing stress by applying exogenous 24-Epibrassinolide (EBR).
Background
Corn (C)Zea mays) As the first large grain crop in China, more than two thirds of the sowing area is mainly distributed in dry farming areas such as northeast, northwest and southwest depending on natural rainfall. Drought and water shortage not only affect the normal growth and development of the corn and interfere the physiological and biochemical metabolism of the corn, but also seriously affect the yield and quality of the corn, and generally, the yield of the corn can be reduced by 20-50% due to drought. The deep-sowing-resistant corn can fully utilize the deep water of the soil, and further can promote the growth of the cornSeeds are fed in to germinate and emerge in time, and the deep-sowing-resistant corn root system is deeper, so that the drought resistance and lodging resistance of the corn can be obviously enhanced. At present, the cultivation of deep-sowing-resistant varieties is one of effective approaches for solving the problems of drought resistance and water conservation in the production of corns.
A great deal of research shows that the length of the mesocotyl and the coleoptile is the main factor for determining the deep sowing resistance of the corn, because the seeds can absorb sufficient water to promote germination when the corn is deeply sowed, and the mesocotyl and the coleoptile can also be obviously elongated so as to provide enough motive power for the seeds to germinate by breaking the soil. Elongation of the mesocotyl and coleoptile is governed not only by their individual physiological properties (POD enzyme activity, H)2O2Content, lignin content, etc.) and is also regulated and controlled by various exogenous hormones (indole-3-acetic acid, IAA, cytokinin, CTK, gibberellin, GA, abscisic acid, ABA, ethylene ethyl, ETH). The studies of Cao Liriong and the like reveal that the hypocotyl of the rice seedling grows along with the increase of the concentrations of the exogenous IAA, CTK and GA in a certain range under the dark condition, wherein the GA treatment has the most obvious response to the hypocotyl of the rice seedling. The research of Zhao Guangwu, etc. shows that the exogenous GA3And IAA treatment, maize mesocotyl cells elongated significantly, but their cell numbers did not change much. Ulrich and Wang have discovered that exogenous IAA promotes the growth of maize germ sheaths and mesocotyl axes in ex vivo conditions. It achieves the effect of cell enlargement mainly by relaxing the epidermal cell wall. Guoguang, etc]Studies indicate exogenous ABA and GA3Has additive effect in promoting the elongation of rice mesocotyl. The Kiyoshi study showed that the simultaneous application of exogenous ETH and GA extended the mesocotyl length of indica rice significantly.
24-Epibrassinolide (EBR) is an important phytosterol hormone and plays an important role in regulating and controlling the growth and development of plants and participating in various stress response processes of the plants. To date, only a few studies have revealed that exogenous EBR can significantly increase the number and size of hypocotyl cells in plants. However, the response mechanism of the exogenous EBR in regulating the corn deep-sowing tolerance under the deep-sowing stress is not clear, and the effect of the exogenous EBR in regulating the corn deep-sowing tolerance cannot be clarified. Therefore, the excellent characteristic of deep sowing of corn seeds and the improvement of the deep sowing and seedling emergence capability of the corn seeds are important measures for ensuring the production safety of the corn in the arid region.
Disclosure of Invention
The technical problem to be solved by the invention is to provide a method for improving the deep-sowing seedling emergence of corn seeds under the deep-sowing stress by applying exogenous 24-Epibrassinolide (EBR). The method can effectively improve the top soil seedling emergence capability of the corn seeds under the deep sowing stress of 15 cm, 20 cm and the like, and can provide reference for guaranteeing the production safety of the corn in the dry region and improving the drought resistance and deep sowing resistance of the corn seeds.
In order to solve the technical problems, the invention adopts the following technical scheme:
a method for improving the deep sowing resistance of corn seeds specifically comprises the following steps:
(1) seed selection: selecting full, uniform and undamaged corn seeds;
(2) preparation of 24-Epinobrassinolide (EBR) solution: using 98% by volume of ethanol and ddH2O EBR was dissolved and diluted to 1.0 and 2.0 mg. L-1The EBR solution (1) of (1), the corresponding EBR solution finally contains 0.1% (v/v) of ethanol and 0.1% (v/v) of Tween-80;
(3) seed disinfection: firstly, disinfecting seeds for 10 min by using NaClO solution with the volume percentage of 0.5 percent, and then, using ddH2O washing the seeds for 3 times, and adsorbing water by using a sterilized filter paper;
(4) seed soaking: soaking the sterilized seeds in 1.0 mg and 2.0 mg of water at room temperature. L-1Soaking seeds in the EBR solution for 24 hours;
(5) preparing a seed germination matrix: mixing sterilized vermiculite with 1.0 and 2.0 mg respectively. L-1The EBR solution is uniformly mixed with soil according to the proportion of 5g to 1mL to prepare a seed germination substrate;
(6) applying 1.0 mg . L-1The method for improving the deep sowing capability of the corn seeds by 15 cm by using the EBR solution comprises the following steps: mixing sterilized vermiculite with 1.0 mg . L-1The EBR solution is uniformly mixed with soil according to the proportion of 5g to 1mL to prepare a seed germination substrate and then is filled into a seed deep-sowing test deviceThen 1.0 mg of . L -130 seeds soaked in the EBR solution are sown in the deep-sowing test device and then covered by 15 cm to contain 1.0 mg . L-1The EBR solution of (a). The test treatment is repeated for 3 times, and 1.0 mg is quantitatively poured every 2 days after sowing . L -140 mL of the EBR solution (1). Artificial climate chamber (12 h of light each day, light intensity 600 mu mol/(s).m2) After germination at day/night temperature (25. + -.1)/(20. + -.1). degree.C, relative humidity 60%) for 10 days, ddH was added2Quickly washing off vermiculite at the root of the seedling by using O water, and adsorbing water by using sterilizing filter paper for measuring corresponding deep-seeding resistance;
(7) applying 2.0 mg . L-1The method for improving the deep sowing capability of the corn seeds by 20 cm by using the EBR solution comprises the following steps: mixing sterilized vermiculite with 2.0 mg . L-1The EBR solution is uniformly mixed with soil according to the proportion of 5g to 1mL to prepare a seed germination substrate, then the substrate is filled into a seed deep-sowing test device with a certain height, and then 2.0 mg of the EBR solution is added . L -130 seeds soaked in the EBR solution are sown in the deep-sowing test device and then covered by 20 cm to contain 2.0 mg . L-1The EBR solution of (a). The test treatment is repeated for 3 times, and 2.0 mg of the fertilizer is quantitatively poured every 2 days after sowing . L -140 mL of the EBR solution (1). Artificial climate chamber (12 h of light each day, light intensity 600 mu mol/(s).m2) After germination at day/night temperature (25. + -.1)/(20. + -.1). degree.C, relative humidity 60%) for 10 days, ddH was added2And (4) rapidly washing off vermiculite at the root of the seedling by using O water, and adsorbing water by using a sterilizing filter paper for measuring the corresponding deep-seeding resistance.
Drawings
FIG. 1 shows the change Rate (RC) of a single trait, the foreign EBR deep-sowing tolerance Regulating Index (RI) of the single trait under corresponding treatment, and the maize deep-sowing tolerance membership value (U) after exogenous EBRs with corresponding concentrations are applied under different deep-sowing stresses;
FIG. 2 is a joint analysis of variance (F-value) of maize inbred line-related traits under different treatments;
wherein the measured values are marked with symbols P or P< 005 or P<The difference at the 0.01 level was significant. RAT emergence rate, MES mesocotyl length, COL coleoptile length, MES + COL sum of mesocotyl and coleoptile, RL root length, SDL seedling length, SOD (MES/COL), SOD activity in mesocotyl/coleoptile, POD (MES/COL), POD activity in mesocotyl/coleoptile, CAT (MES/COL), CAT activity in mesocotyl/coleoptile, APX (MES/COL), APX activity in mesocotyl/coleoptile, H2O2(MES/COL) mesocotyl/coleoptile H2O2Content, LIG (MES/COL), lignin content in mesocotyl/coleoptile. The same applies below. FIG. 3 shows the effect of exogenous EBR on 6 phenotypic traits of maize inbred lines under different deep-sowing treatments;
FIG. 4 is a graph showing the effect of exogenous EBR on the activity of 4 antioxidant enzymes in the mesocotyl and coleoptile of maize inbred lines under different deep-sowing treatments;
FIG. 5 shows H in hypocotyls and coleoptiles of inbred lines of maize after exogenous EBR application under different deep-sowing treatments2O2Influence of lignin content;
FIG. 6 is the correlation analysis between different traits of inbred lines of maize under different treatments;
FIG. 7 is a cluster analysis between two types of depth-of-cut treatments (15, 20 cm) between groups with different concentrations of exogenous EBR applied;
FIG. 8 is a deep sowing modulation index (RI) of the traits associated with different maize inbred lines after exogenous EBR was applied under the stress of 15 cm deep sowing;
wherein RI 1-RI 18 respectively refer to emergence rate, mesocotyl length, embryo sheath length, sum of mesocotyl and coleoptile, root length, seedling length, SOD in mesocotyl, POD, CAT, APX enzyme activity, H2O2Lignin content, SOD, POD, CAT, APX enzyme activity in coleoptile, H2O2And a deep-seeding resistance adjustment index (RI) of lignin content. The same applies below.
FIG. 9 is a deep sowing modulation index (RI) of the traits associated with different maize inbred lines after exogenous EBR was applied under 20 cm deep sowing stress;
FIG. 10 is a comprehensive evaluation of deep-sowing tolerance of maize inbred lines after exogenous EBR is applied under different deep-sowing stresses;
FIG. 11 is an overall view of a seed deep-sowing testing apparatus;
FIG. 12 is a schematic top view of a seed deep-sowing testing apparatus;
FIG. 13 is an overall view of a gear type seed deep-sowing testing device;
FIG. 14 is a partial schematic view of a gear type seed deep-sowing testing device;
FIG. 15 is an overall view of a chuck type deep-seeding test device;
fig. 16 is an overall schematic diagram of a pulley type seed deep-sowing testing device.
In fig. 11-16, a sowing depth measuring cylinder 1, a sowing depth measuring scale 2, a substrate tray 3, a label groove 4, a clamping strip 5, a screw 6, a fixing clamp ring 7, a rotating bottom support 8, a motorized gear 9, a handle 10, a gear fixer 11, a linkage gear 12, a chain 13, a clamping groove 14, a wire passing pipe 15, a buckle 16, a pulley 17 and a rope 18.
Detailed description of the invention
The methods and devices used in the following examples of the present invention are conventional methods and devices unless otherwise specified; the equipment and the reagent are all conventional equipment and reagents purchased by a reagent company. In order to make the objects, technical solutions and advantages of the present invention more apparent, the following detailed description of the embodiments of the present invention is provided in connection with the specific embodiments. Examples of these preferred embodiments are illustrated in the specific examples.
It should be noted that, in order to avoid obscuring the technical solutions of the present invention with unnecessary details, only the technical solutions and/or processing steps closely related to the technical solutions of the present invention are shown in the embodiments, and other details that are not relevant are omitted.
Example 1
The invention provides a screening method for improving the optimal parameters of the deep sowing resistance of corn seeds, which comprises the following steps:
1. collecting materials: 2 parts of the inbred lines K12 and Ji 853 of the intolerant and deep-sown corn are core corn germplasm materials commonly used in China, wherein K12 is derived from descendants of Huang Zao SixWeichun, Ji 853 is derived from descendants of (Huang Zao Sixfrom 330) xHuang Zao Sio, and the 2 parts of inbred line seeds are provided by molecular marker assisted breeding laboratories in the drought habitat crop science of Gansu province.
2.24-preparation of Epinobrassinolide (EBR) solution: EBR was purchased from Sigma, USA and has the molecular formula C28H48O6. Ethanol and ddH with the volume percentage of 98 percent before use2O dissolution and dilution of EBR to 5 different concentrations (0 mg). L-1(i.e., ddH)2O）、0.5 mg . L-1、1.0 mg . L-1、1.5 mg . L-1And 2.0 mg. L-1) EBR solutions, corresponding to EBR solutions, finally contained 0.1% (v/v) ethanol and 0.1% (v/v) Tween-80. And (5) standby.
3. Seed selection and disinfection: selecting K12 and Ji 853 seeds with full, uniform and undamaged seeds, sterilizing the seeds for 10 min with 0.5 vol% NaClO solution, and then using ddH2The seeds were rinsed 3 times with O and the adhering water was blotted dry with sterile filter paper. And (5) standby.
4. Seed soaking: the sterilized K12 and Ji 853 seeds are respectively soaked in 5 EBR solutions with corresponding concentrations for 24 h at room temperature. And (5) standby.
5. Preparing a seed germination matrix: the sterilized vermiculite and 5 kinds of EBR solutions with corresponding concentrations are uniformly mixed with soil according to the proportion of 5g to 1mL to prepare the seed germination substrate. And (5) standby.
6. Seed deep sowing stress test: the corresponding seed germination matrixes are respectively filled into a certain height of any device (with the height of 50 cm and the diameter of 17 cm) described in the embodiment 4-8, 30 seeds of K12 and Ji 853 soaked in different EBR solutions are sown into any device described in the embodiment 4-8, and then the corresponding seed germination matrixes are respectively covered by 3 cm, 15 cm and 20 cm. The assay was set to 11 treatments, namely positive normal control treatment CK (+): 3 cm sowing depth +0 mg. L-1 EBR（ddH2O), negative deep-seeding stress control treatment CK (-)1/CK (-) 2: 15/20 cm sowing depth +0 mg. L-1 EBR（ddH2O), the stress treatment for the deep sowing of 15 cm is respectively as follows: s1-1 (15 cm sowing depth +0.5 mg). L-1 EBR）S1-2 (15 cm sowing depth +1.0 mg). L-1EBR), S1-3 (15 cm depth of seeding +1.5 mg). L-1EBR), S1-4 (15 cm depth of seeding +2.0 mg. L-1EBR), 20 cm deep-seeding stress treatment were: s2-1 (20 cm sowing depth +0.5 mg). L-1EBR), S2-2 (20 cm depth of cut +1.0 mg. L-1EBR), S2-3 (20 cm depth of seeding +1.5 mg). L-1EBR), S2-4 (20 cm depth of seeding +2.0 mg. L-1EBR). Each test treatment was repeated 3 times, and 40 mL of EBR solution of the corresponding concentration was quantitatively poured every 2 days after sowing. Artificial climate chamber (12 h of light each day, light intensity 600 mu mol/(s).m2) After germination at day/night temperature (25. + -.1)/(20. + -.1). degree.C, relative humidity 60%) for 10 days, ddH was added2And (4) rapidly washing off vermiculite at the root of the seedling by using O water, and adsorbing water by using a sterilizing filter paper for measuring the corresponding deep-seeding resistance.
7. Determination of deep-seeding resistance: determining the phenotypic traits such as emergence Rate (RAT), mesocotyl length (MES), coleoptile length (COL), the sum of mesocotyl and coleoptile length (MES + COL), seedling length (SDL), Root Length (RL) and the like under each treatment; taking the mesocotyl and coleoptile of each inbred line after corresponding treatment, and determining H in the mesocotyl and coleoptile by adopting a titanium sulfate colorimetric method2O2Content (H)2O2 content in mesocotyl/coleoptile, H2O2(MES/COL) measuring the content of lignin in the mesocotyl and coleoptile by Syros method (LIG/MES/COL), measuring superoxide dismutase (SOD) in the mesocotyl and coleoptile by the nitro blue tetrazolium photochemical reduction method (MES/COL)), measuring peroxidase in the mesocotyl and coleoptile by the guaiacol method (POD/MES), measuring catalase in the mesocotyl and coleoptile by the ultraviolet absorption method (CAT/COL), ascorbate peroxidase (as)Physiological properties such as sorbate peroxidase in mesotype/coleoptile, APX (MES/COL)).
8. Rate of change of individual traits with corresponding treatment: the Rate of Change (RC) of a single trait under deep-sowing stress and RC after applying an exogenous EBR under deep-sowing stress are calculated according to the formulas (1) and (2) in the attached figure 1. In the formula: RC (resistor-capacitor) capacitoriCThe rate of change of the corresponding trait under the i-th deep-sowing stress, RCinThe change rate of the corresponding traits after applying EBR with the nth concentration under the ith deep-sowing stress, TCK(+)Measured under control treatment CK (+) for normal 3 cm sowing depth, TCK(-)iControl treatment of the measurement under CK (-) i for negative ith deep-planting stress (15 or 20 cm), Ti-nThe measured value of the corresponding trait at the nth concentration under the ith deep-sowing stress.
9. Exogenous EBR deep-seeding resistance modulation index of individual trait: in order to conveniently evaluate the deep-sowing resistance of the exogenous EBR with corresponding concentration in the single character of the corn, the invention newly defines the regulation index of deep-sowing Resistance (RI) of the exogenous EBR with the single character according to the measured character values after the exogenous EBR with corresponding concentration is applied under different deep-sowing stresses, and the formulas are shown as the formula (3) and the formula (4) in the attached figure 1. In the formula: RI (Ri)inThe corresponding traits of the ith deep-sowing stress are shown in the
Figure DEST_PATH_IMAGE001
And (3) calculating the exogenous EBR deep-seeding resistance regulation index under the seed concentration by adopting the formula (3) if the measured character is in positive correlation with the deep-seeding resistance of the corn, and otherwise, calculating by adopting the formula (4). The positive RI value indicates that the exogenous EBR with the corresponding concentration plays a positive regulation role on the deep sowing stress, and the negative RI value indicates that the exogenous EBR with the corresponding concentration plays a negative regulation role on the deep sowing stress.
10. Comprehensively evaluating the deep sowing resistance of the corn after applying the exogenous EBR: taking RI value of each measured character as evaluation index of maize deep sowing resistance strength, adopting membership function method to comprehensively evaluate maize deep sowing resistance strength membership value (U) after applying exogenous EBR with corresponding concentration under different deep sowing stresses, wherein the formulas are shown as formula (5) and formula (U) in figure 1(6). In the formula: u shapeinThe corresponding characters under the ith deep sowing stress are subjected to the exogenous EBR deep sowing tolerance membership value, RI, of the nth concentrationiminThe corresponding traits of the ith deep-sowing stress are shown in the
Figure 910320DEST_PATH_IMAGE001
Minimum RI value at species concentration, otherwise, RIimanThe corresponding traits of the ith deep-sowing stress are shown in the
Figure 827461DEST_PATH_IMAGE001
Maximum RI value at seed concentration. If the measured RI is positively correlated with the deep-seeding resistance regulation effect after the external source EBR is applied, the formula (5) is adopted for calculation, otherwise, the formula (6) is adopted for calculation, RI membership values of the external source EBR applied with various concentrations under the corresponding deep-seeding stress are accumulated, the arithmetic mean of the RI membership values is calculated for comparison, and the larger the value is, the stronger the deep-seeding resistance of the corn after the external source EBR is applied is.
11. And (4) analyzing results: all experimental data were statistically plotted using Microsoft Excel 2016 software, and Duncan multiple comparison analysis of variance, Pearson correlation analysis, and inter-group system clustering analysis using IBM SPSS 19.0 software.
Example 2
The invention provides a screening result for improving a parameter of deep sowing resistance of corn seeds, which comprises the following specific screening results:
1. and (3) carrying out differential analysis on phenotype and physiological characteristics of the maize inbred line after applying exogenous EBR under different deep-sowing treatments: the joint variance analysis of 18 related traits of 2 maize inbred lines under 11 different treatments, such as CK (+), CK (-)1, CK (-)2, S1-1-S1-4 and S2-1-S2-4, shows that the correction models of the 18 related traits all reach the level shown in figure 2 (Table 1)P <0.05 or P <The difference at the 0.01 level was significant. It was demonstrated that the overall variance of these 18 traits was significant in the combined analysis, and therefore it was effective to perform variance analysis on the inbred lines, the sowing depth, the EBR concentration and the interaction thereof for these 18 traits. Further analysis shows that most characters are in inbred line, sowing depth, EBR concentration, inbred line and sowing depth interaction, inbred line and EBR concentrationThe difference between the density interaction, the sowing depth multiplied by the EBR concentration interaction and the self-bred line multiplied by the sowing depth multiplied by the EBR concentration interaction all reachP <0.05 or P <0.01 significant level. The characteristics of the maize inbred line are obviously influenced by the genetic basis of the maize genotype, the sowing depth and the EBR concentration, but the influence degree of the characteristics of the maize inbred line is different. Wherein the sowing depth has the maximum influence on the emergence rate, the germ sheath length, the activities of POD and APX in mesocotyl and the POD enzyme in the germ sheath, the self-bred line has the second influence on the 5 characters, the EBR concentration has the minimum influence on the 5 characters,Fthe values decrease in sequence. The sowing depth is adjusted to the mesocotyl length, the sum of the mesocotyl and coleoptile, the SOD and CAT enzyme activities in the mesocotyl, and H2O2And lignin content and SOD enzyme activity in coleoptile, H2O2The influence degree of the content of the lignin is the largest, the influence degree of EBR concentration on the 9 characters is the second order, the influence degree of the inbred line on the 9 characters is the smallest,Fthe values decrease in sequence. Indicating that the sowing depth can significantly affect the 14 traits of the corn genotype, and the traits of different corn genotypes change to the greatest extent with the increase of the sowing depth. Secondly, the inbred line has the greatest influence on the emergence rate, the sowing depth has the next influence on the emergence rate, the EBR concentration has the smallest influence on the emergence rate,Fthe values decrease in sequence. The emergence rate is controlled to be the maximum by the genetic basis of the maize genotype, so that the emergence rate is influenced to be the maximum by the self-bred line in the evaluation of the deep sowing tolerance of the maize. In addition, the EBR concentration has the greatest influence on the activity of CAT and APX enzymes in root length and coleoptile, the sowing depth has the second influence on the 3 characters, the inbred line has the least influence on the 3 characters,Fthe values decrease in sequence. Suggesting that the deep sowing stress treatment of different genotype maize inbred lines by different concentrations of EBR can have the greatest influence on the genotype root length of maize and the activity of CAT and APX enzymes in coleoptile.
2. Influence of different deep-sowing treatments on phenotype and physiological characteristics of maize inbred lines: compared with CK (+) processed by the control of sowing depth of normal 3 cm in the positive direction, the maize inbred lines of the Gji 853, K12 and the like have the mesocotyl length and the embryo length under the control of sowing depth of 15 cm and 20 cm in the negative direction and under the CK (-)1/CK (-)2 and are in the control of sowing depth of 10 cm and 20 cm in the positive directionLength of sheath, sum of mesocotyl and coleoptile, length of root, SOD and POD activity in mesocotyl/coleoptile, H2O2And lignin content, which are increased by 115.0/139.4%, 30.1/45.5%, 68.2/87.6%, 17.7/42.8%, 18.4/28.9% and 29.7/46.0%, 81.0/125.1% and 66.4/153.7%, 21.0/70.3% and 89.9/292.6%, 228.0/392.4% and 185.6/291.6%, respectively, while emergence rate, seedling length, CAT and APX activities in mesocotyl and coleoptile are decreased by 89.2/100.0%, 28.7/44.3%, 22.2/30.2% and 9.2/14.4%, 89.0/92.9% and 70.5/74.2% (FIGS. 3, 4 and 5). In addition, compared with the sowing depth of 3 cm, the average change rate RC of the 18 characters under the deep sowing of 15 cm is-228.0% (the lignin content in the mesocotyl axis) to 89.2% (the emergence rate), and the average RC of the 18 characters under the deep sowing of 20 cm is-392.4% (the lignin content in the mesocotyl axis) to 100.0% (the emergence rate).
3. The change of phenotype and physiological characteristics of the maize inbred line after applying exogenous EBR under different deep-sowing treatments: 15. after exogenous EBR is applied under the deep sowing stress of 20 cm, the mesocotyl, the coleoptile, the root length and the seedling growth of the Ji 853 and K12 can be effectively improved, so that the emergence rates of the Ji 853 and K12 under the corresponding deep sowing stress are obviously improved, but the exogenous EBR with different concentrations has different influence degrees on various phenotypic traits of the maize inbred line. As can be seen from FIG. 3, 1.0 mg was applied under the stress of 15 cm of seeding depth. L-1After EBR, the emergence rate, the mesocotyl length, the coleoptile length, the sum of the mesocotyl and the coleoptile and the seedling length of 2 maize inbred lines are the maximum, the average of the emergence rate, the mesocotyl length, the coleoptile length, the mesocotyl and coleoptile length and the seedling length are respectively 1.0 time and 9.4 times, 3.1 time and 1.4 times, 1.5 time and 1.2 times, 2.2 time and 1.3 times and 0.9 time and 1.3 times of CK (+) and CK (-)1, the average RC of the seedling length is between 912.4 percent (emergence rate) and 21.0 percent (coleoptile length), and 2.0 mg of the seedling. L-1After EBR of (1), 2 maize inbred lines had the longest root length, which was 1.0 and 0.9 times the average of CK (+) and CK (-)1, respectively, and the average RC was 14.7%. 0.5 mg under the stress of 20 cm sowing depth. L-1After EBR of (1.0 mg), the germ sheath length and root length of 2 maize inbred lines were maximal, with average 1.4 and 1.0 times, 1.1 and 0.8 times CK (+) and CK (-)1, respectively, and average RC of 10.4 and 26.7%, applying 1.0 mg. L-1After EBR of (2 parts of corn)The emergence rate and the seedling length of the cross line were the greatest, with average 0.7 and 19.7 times, 0.7 and 1.2 times of CK (+) and CK (-)2, respectively, and the average RC of the seedling length was-44.7%, 2.0 mg applied. L-1After EBR of (1), the mesocotyl length and the sum of mesocotyl and coleoptile were the largest for 2 maize inbred lines, which averaged 3.5 and 1.6 times, 2.3 and 1.3 times for CK (+) and CK (-)2, respectively, and their averaged RC was-80.1 and-46.7%, respectively.
After exogenous EBR is applied under two deep sowing stresses, the activity of various antioxidant enzymes in the hypocotyl and the coleoptile of the maize inbred line can be obviously changed, but the influence degree of the exogenous EBR with different concentrations on the activity of various antioxidant enzymes is different. As can be seen from FIG. 4, 1.0 mg was applied under the stress of 15 cm of seeding depth. L-1After EBR (E-Bl-Cohn, maize) of 2 maize inbred lines, the activity of SOD, CAT and APX enzymes in the hypocotyl/coleoptile is the largest, the average activity is 1.4/1.8, 1.2/1.4 times, 0.9/1.0, 1.2/1.1 times, 0.6/2.6 and 5.5/8.8 times of CK (+) and CK (-)1 respectively, the average change rate is-802.7% (APX enzyme activity in coleoptile) to-14.1% (CAT enzyme activity in coleoptile), 0.5 mg is added. L-1After EBR of (1.5 mg), POD enzyme activity was greatest in the mesocotyl of 2 maize inbred lines, which were 2.2 and 1.2 times as high as CK (+) and CK (-)1, respectively, with an average RC of-20.2%, applied. L-1After EBR of (1), POD enzyme activity was maximal in coleoptiles of 2 maize inbred lines, which were on average 2.0 and 1.2 times that of CK (+) and CK (-)1, respectively, with an average RC of-37.0%. Applying 1.0 mg under the stress of 20 cm sowing depth. L-1After EBR (E-Bl-Cohn), the SOD and POD enzyme activities were maximal in the mesocotyl/coleoptile of 2 maize inbred lines, which were 1.4/1.8 and 1.2/1.4 times, 2.2/2.4 and 1.0/1.2 times, respectively, of CK (+) and CK (-)2, and the average change rate was-23.5% (SOD enzyme activity in coleoptile) to-1.5% (POD enzyme activity in mesocotyl), and 2.0 mg was applied. L-1After EBR, the activity of CAT and APX enzymes in the mesocotyl/coleoptile of 2 maize inbred lines is the maximum, the average activity is 1.1/1.1, 1.4/1.3 times, 2.1/3.3 and 10.4/13.1 times of CK (+) and CK (-)2 respectively, and the average RC is-3219.3% (the activity of APX enzyme in the mesocotyl) to-32.3% (the activity of CAT enzyme in the mesocotyl).
Can obviously influence the maize inbred line after applying exogenous EBR under different deep sowing stressesH in mesocotyl and coleoptile2O2And lignin accumulation, but different concentrations of exogenous EBR for maize inbred line H2O2And the amount of lignin accumulated, to a different extent. As can be seen from FIG. 5, 0.5 mg was applied under the stress of 15 cm of seeding depth. L-1After EBR of (1), H in the mesocotyl of 2 parts of maize inbred line2O2The minimum content, which is on average 0.9 and 0.8 times CK (+) and CK (-)1, respectively, and an average RC of 25.6%, was applied at 1.5 mg. L-1After EBR of (1.0 mg), the lignin content in the hypocotyls of 2 maize inbred lines was minimal, which was 1.1 and 0.3 times as high as CK (+) and CK (-)1, respectively, and the average RC was 66.9%, applying 1.0 mg. L-1After EBR, 2 parts of H in coleoptile of maize inbred line2O2And lignin content was minimal, with average 1.2 and 0.6 times, 1.1 and 0.4 times that of CK (+) and CK (-)1, respectively, and with average RC of 33.0% and 59.6%. Applying 1.5 mg under the stress of 20 cm sowing depth. L-1After EBR of (1), H in the mesocotyl of 2 parts of maize inbred line2O2Lignin content and H in coleoptile2O2The contents were all the smallest, which were 1.0 and 0.7 times, 1.2 and 0.3 times, 1.8 and 0.6 times as large as CK (+) and CK (-)2, respectively, and the average RC thereof was 35.4, 72.3 and 40.9%, 1.0 mg was applied. L-1After EBR of (1), the lignin content in coleoptile of maize inbred line is minimum, which is 1.3 and 0.4 times of CK (+) and CK (-)1 respectively on average, and the average RC is 54.1%.
4. Correlation analysis among various traits of the maize inbred line under different deep sowing treatments: the correlation analysis of the 18 traits of 2 maize inbred lines under the deep sowing treatment of 3 cm, 15 cm and 20 cm shows that the emergence rate of the 3 maize inbred lines under the deep sowing is very significant with the sum of mesocotyl length, embryo sheath length, mesocotyl and coleoptile, the activities of SOD, CAT and APX enzymes in the mesocotyl and the activities of SOD, CAT and APX enzymes in the coleoptile as shown in the attached figure 6 (table 2) ((the activities of SOD, CAT and APX enzymes in the mesocotyl are all significantP <0.01) positive correlation with root length and seedling length (a)P <0.05) positive correlation with POD enzyme activity in mesocotyl/coleoptile, H2O2And the lignin content are in extremely obvious negative correlation. It is stated that the mesocotyl length and coleoptile of the maize were obtained under these 3 deep-sowing treatmentsLength, sum of mesocotyl and coleoptile, root length, seedling length, SOD, POD, CAT and APX enzyme activities in mesocotyl/coleoptile, H2O2And the lignin content play an important role in the emergence of the corn seedlings, and the corn seedlings are sent out of the ground surface by the synergistic effect of the lignin content and the corn seedlings. In addition, the sowing depth, the length of mesocotyl, the sum of mesocotyl and coleoptile, the root length, SOD in mesocotyl/coleoptile, the activity of POD enzyme, and H2O2Has obvious positive correlation with the lignin content, obvious positive correlation with the germ sheath length and extremely obvious negative correlation with the rest 6 individual characters. The deep sowing can obviously affect the 18 characters of the maize inbred line, so the deep sowing can be used as an important identification index for evaluating the deep sowing tolerance of the maize inbred line under different deep sowing treatments.
5. And (3) performing deep sowing resistance clustering evaluation on the maize inbred line after applying exogenous EBR under different deep sowing stresses: applying 0.5-2.0 mg of 18 related traits to normal 3 cm sowing depth control treatment CK (+), negative 15/20 cm deep sowing stress control treatment CK (-)1/CK (-)2 and deep sowing 15/20 cm under stress. L-1Systematic clustering analysis among groups under EBR treatment S1-1-S1-4/S2-1-S2-4 shows (figure 7), under the deep-sowing stress of 15 cm, 6 different corresponding treatments are clustered into 2 groups of S1-3, CK (-)1, S1-4 and the like and CK (+), S1-2, S1-1 and the like when the Euclidean distance is 7.5, which indicates that 0.5 and 1.0 mg are applied under the deep-sowing stress of 15 cm. L-1The maize inbred line after EBR has stronger deep sowing resistance. Under the 20 cm deep sowing stress, 6 different corresponding treatments are clustered into 3 groups of S2-4, S2-3, S2-2, S2-1 and the like, CK (+) and the like and CK (-)2 and the like when the Euclidean distance is 7.5, which indicates that 0.5-2.0 mg of the treatment is applied under the 20 cm deep sowing stress. L-1The deep sowing tolerance of the maize inbred line after EBR is CK (+) -CK (-) 2.
6. The deep sowing tolerance adjusting index of each character of the maize inbred line after applying exogenous EBR under the deep sowing stress: 15. after the maize inbred line is applied with exogenous EBR with different concentrations under the 20 cm deep sowing stress, the deep sowing resistance regulation directions and sizes of 18 related traits of the maize inbred line are obviously different, and the RI values are different, as shown in the attached figures 8 (table 3) and 9 (table 4). 0.5 mg was applied under deep-sowing stress of 15 cm. L-1EBR 2 parts maize inbredThe mean RI value for the root length was-0.287 max, 1.5 mg was applied. L-1The average RI value of the lignin content in the mesocotyl of 2 parts of maize inbred lines after EBR is 0.964 at most, and the RI values of other characters are all applied by 1.0 mg. L-1The average RI of the EBR of (1) is-1.144-5.772, as shown in FIG. 8 (Table 3). In addition, 1.5 mg was applied under deep-sowing stress of 20 cm. L-1The average RI value of the germ sheath length of 2 parts of maize inbred lines after EBR is 0.067 at most, 2.0 mg is added. L-1The RI values of the rest 17 characters of the maize inbred lines of 2 parts after EBR are the maximum, and the average RI value is-0.505-6.697, as shown in figure 9 (Table 4). However, the method can only qualitatively and fuzzinly research the optimal regulation effect of the exogenous EBR on various adversity stresses, but cannot comprehensively and quantitatively reveal the optimal regulation effect of the exogenous EBR on various adversity stresses.
7. And (3) comprehensively evaluating the deep sowing resistance of the maize inbred line after applying exogenous EBR under different deep sowing stresses: based on the consideration, the RI values of 18 related characters of the corn after applying the exogenous EBR with different concentrations under the deep sowing treatment of 15 cm and 20 cm are taken as the comprehensive evaluation index of the deep sowing resistance of the corn, and a membership function method is adopted to quantitatively and scientifically, objectively and comprehensively evaluate the deep sowing resistance of the corn after applying the exogenous EBR with different concentrations under the deep sowing stress of 15 cm and 20 cm. The deep sowing resistance of the maize inbred line after applying exogenous EBR under the stress of 15 cm deep sowing tends to increase and decrease along with the increase of the EBR concentration, and 1.0 mg is applied under the stress of 15 cm deep sowing. L-1After EBR, the maize inbred line has the strongest deep sowing resistance, the membership value U of the deep sowing resistance is 0.560, and 0.5 mg is applied. L-1After EBR of (1), the maize inbred line had the weakest deep-planting resistance and U was 0.420, as shown in FIG. 10 (Table 5). The deep sowing resistance of the maize inbred line after applying exogenous EBR under the 20 cm deep sowing stress is increased along with the increase of the EBR concentration, and 2.0 mg is applied under the 20 cm deep sowing stress. L-1After EBR (E-Back-propagation) of (1), the maize inbred line has the strongest deep-seeding resistance, U is 0.672, and 0.5 mg is applied. L-1The maize inbred line had the weakest deep-planting tolerance after EBR (Takara Shuzo), with a U of 0.351, as shown in FIG. 10 (Table 5).
As can be seen from the above examples, the method not only reveals the influence of the 15 cm and 20 cm deep sowing stress on the form and physiological characteristics of the deep sowing emergence of the corn seeds, but also newly defines the exogenous EBR deep sowing resistance index RI with a single character, takes the RI as the comprehensive deep sowing resistance evaluation index, adopts a membership function method, scientifically, objectively and comprehensively evaluates the regulating effect of applying exogenous EBRs with different concentrations on the deep sowing resistance of the corn seeds under the 15 cm and 20 cm deep sowing stress, lays a theoretical foundation for improving the deep sowing resistance characteristics of the corn seeds, provides technical support for the drought resistance of the corn sowed in the seedling form establishment stage, and has high application value.
Example 3
The invention provides a method for improving the deep sowing resistance of corn seeds, which comprises the following steps:
the optimal parameters for improving the deep sowing resistance of the corn seeds by applying the EBR solution under the deep sowing stress of 1.15 cm are as follows: the deep sowing resistance of the maize inbred line after applying exogenous EBR under the stress of 15 cm deep sowing tends to increase and decrease along with the increase of the EBR concentration, and 1.0 mg is applied under the stress of 15 cm deep sowing. L-1After EBR, the maize inbred line has the strongest deep sowing resistance, and the deep sowing resistance membership value U is 0.560.
The optimal parameters for improving the deep sowing resistance of the corn seeds by applying the EBR solution under the deep sowing stress of 2.20 cm are as follows: the deep sowing resistance of the maize inbred line after applying exogenous EBR under the 20 cm deep sowing stress is increased along with the increase of the EBR concentration, wherein 2.0 mg is applied under the 20 cm deep sowing stress. L-1After EBR, the maize inbred line has the strongest deep sowing resistance, and U is 0.672.
Example 4
The embodiment provides a rotary type seed deep-sowing testing device, as shown in fig. 11 and 12, the structure of which comprises:
the middle part of a rotating bottom support 8 is connected with a screw rod 6, a sowing depth quantifying cylinder 1 is sleeved above the rotating bottom support 8, a fixing clamping ring 7 is embedded in a groove of the screw rod 6 and is fixed inside the sowing depth quantifying cylinder 1, clamping strips 5 are arranged on two opposite sides inside the sowing depth quantifying cylinder 1, a screw hole in the middle part of a substrate tray 3 penetrates through the screw rod 6, and a groove matched with the clamping strips 5 is formed in the outer side of the substrate tray 3; a seeding depth measuring scale 2 and a label groove 4 are arranged outside the seeding depth measuring cylinder 1.
Further, the screw rod 6 is connected the top of rotatory collet 8 department and is equipped with the recess, and fixed rand 7 is inlayed in the recess for connect fixed broadcast deep quantitative section of thick bamboo 1 and rotatory collet 8.
Furthermore, the sowing depth quantifying cylinder 1 is made of transparent polyvinyl chloride or PMMA material, and a plurality of small holes are uniformly formed outside the sowing depth quantifying cylinder 1 so as to conveniently remove redundant water in the sowing depth quantifying cylinder 1;
further, the sowing depth measuring scale 2, the substrate tray 3, the screw 6 and the rotating bottom support 8 are all made of stainless steel materials;
furthermore, 2-4 clamping strips 5 are arranged on two opposite sides in the sowing depth quantifying cylinder 1 and used for fixing the substrate tray 3, so that the screw 6 can only drive the substrate tray 3 to ascend and descend without rotating when rotating;
further, the rotating bottom support 8 drives the screw rod 6 to rotate, so that the substrate tray 3 is lifted, and the seed sowing depth is adjusted at any time.
Example 5
The embodiment provides a gear type seed deep-sowing testing device, as shown in fig. 13 and 14, the structure thereof includes:
the seeding depth quantifying barrel 1 is respectively provided with a motorized gear 9 from top to bottom, the motorized gear 9 is connected through a chain 13, the two sides of the seeding depth quantifying barrel 1 are provided with bar-shaped windows for the chain 13 to move, a matrix tray 3 is arranged in the seeding depth quantifying barrel 1, a linkage gear 12 is sequentially arranged above the motorized gears 9 on the two opposite sides of the outside of the matrix tray 3, one side of the motorized gear 9 is connected with a gear fixing device 11, a handle 10 is connected to the outside of the linkage gear 12, and a seeding depth quantifying ruler 2 and a label slot 4 are arranged outside the seeding depth quantifying barrel 1.
Furthermore, the sowing depth quantifying cylinder 1 is made of transparent polyvinyl chloride or PMMA material, and a plurality of small holes are uniformly formed outside the sowing depth quantifying cylinder 1 so as to conveniently remove redundant water in the sowing depth quantifying cylinder 1;
furthermore, the sowing depth measuring scale 2, the substrate tray 3, the handle 10 and the gear fixer 11 are all made of stainless steel materials;
further, the chain 13 is a MURTFELDT chain;
further, the motorized gear 9 and the linkage gear 12 both adopt POM gears;
further, the rotating handle 10 drives the motorized gears 9 on two opposite sides outside the substrate tray 3 to rotate along with the chain 13, so that the substrate tray 3 is lifted, and the seeding depth of seeds is adjusted at any time.
Example 6
This embodiment provides a draw-in groove formula seed deep-sowing test device, as shown in fig. 15, its structure includes:
the opposite two sides of the sowing depth quantifying barrel 1 are respectively provided with a clamping groove 14, the inside of the sowing depth quantifying barrel 1 is provided with a substrate tray 3, the opposite two sides of the outside of the substrate tray 3 are respectively provided with a handle 10, the other side of the handle 10 penetrates through the clamping groove 14, and the outside of the sowing depth quantifying barrel 1 is provided with a sowing depth quantifying ruler 2 and a label groove 4.
Furthermore, the sowing depth quantifying cylinder 1 is made of transparent polyvinyl chloride or PMMA material, and a plurality of small holes are uniformly formed outside the sowing depth quantifying cylinder 1 so as to conveniently remove redundant water in the sowing depth quantifying cylinder 1;
furthermore, the sowing depth measuring scale 2, the substrate tray 3 and the handle 10 are all made of stainless steel materials;
furthermore, parameters of the clamping grooves 14 on two opposite sides of the sowing depth quantifying cylinder 1 are preferably set to be a pair every 1 cm, and the clamping grooves 14 are made of stainless steel materials;
further, the handle 10 drives the substrate tray 3 to be clamped to the corresponding height of the clamping groove 14, so that the substrate tray 3 is lifted, and the seed sowing depth is adjusted at any time.
Example 7
The present embodiment provides a pulley type deep-sowing testing device for seeds, as shown in fig. 16, the structure thereof includes:
buckle 16 is respectively established to the opposite both sides about 1 outside of the deep quantitative section of thick bamboo of broadcasting, and spool 15 is respectively inlayed to the opposite both sides about 1 inside of the deep quantitative section of thick bamboo of broadcasting, broadcasts deep quantitative section of thick bamboo 1 inside and establishes matrix tray 3, and pulley 17 is respectively established to 3 outside opposite both sides of matrix tray, and pulley 17 is passed to rope 18, and on the spool 15 was passed respectively and is fixed in buckle 16 in 18 both sides of rope, deep quantitative section of thick bamboo 1 outside of broadcasting still set up deep quantitative chi 2 and label groove 4.
Furthermore, the sowing depth quantifying cylinder 1 is made of transparent polyvinyl chloride or PMMA material, and a plurality of small holes are uniformly formed outside the sowing depth quantifying cylinder 1 so as to conveniently remove redundant water in the sowing depth quantifying cylinder 1;
furthermore, the sowing depth measuring scale 2, the substrate tray 3 and the handle 10 are all made of stainless steel materials;
further, the pulley 17 is an NSK pulley;
furthermore, 2-4 buckles 16 and the wire passing pipes 15 are arranged, and the buckles 16 and the wire passing pipes 15 can be preferably detached;
further, on being fixed in buckle 16 of one side about the quantitative section of thick bamboo 1 of broadcasting deeply 18 one side, the buckle 16 of the opposite side about the quantitative section of thick bamboo 1 of broadcasting deeply loosens pulling rope 18, drives pulley 17 and removes, makes matrix tray 3 realize going up and down, and then adjusts the seed depth of planting at any time.
Example 8
The embodiment provides a use method of a seed deep-sowing testing device,
when the device is used, firstly, a matrix tray 3 (the matrix tray 3 is preferably made of stainless steel materials, the height of the matrix tray 3 is preferably set to be 5 cm) in the deep sowing quantifying cylinder 1 is adjusted according to a deep sowing quantifying cylinder 1 (the deep sowing quantifying cylinder 1 is preferably made of transparent polyvinyl chloride or PMMA materials, and a plurality of small holes are uniformly formed outside the deep sowing quantifying cylinder 1 so as to conveniently remove redundant water in the deep sowing quantifying cylinder 1, the parameters are preferably set to be 50 cm in height, 15-25 cm in inner diameter and preferably 17 cm in depth, and the depth of the deep sowing quantifying cylinder 1 is preferably set to be 2 (the depth of the deep sowing quantifying cylinder 2 is preferably made of stainless steel materials), and particularly, a rotary bottom support 8 (the rotary bottom support 8 is preferably made of stainless steel materials) drives a screw 6 (the screw 6 is preferably made of stainless steel materials) to rotate; or the handle 10 is rotated (the handle 10 is preferably made of stainless steel material) to drive the motor-driven gears 9 (the motor-driven gears 9 are preferably POM gears) on two opposite sides outside the substrate tray 3 to rotate to corresponding positions along with the chain 13 (the chain 13 is preferably made of MURTFELDT chains), and the gear holders 11 on one side of the motor-driven gears 9 on two opposite sides outside the substrate tray 3 fix the linkage gear 12 (the linkage gear 12 is preferably made of POM gears); or the handle 10 drives the substrate tray 3 to be clamped to the corresponding height of the clamping grooves 14 (the parameters of the clamping grooves 14 are preferably set as a pair every 1 cm, and the clamping grooves 14 are preferably made of stainless steel materials); or the rope 18 passes through the wire-passing pipes 15 (2-4 are arranged on the wire-passing pipes 15, the wire-passing pipes 15 can be preferably disassembled) and is fixed on the buckles 16 at the upper and lower opposite sides outside the seeding depth quantifying barrel 1 (2-4 are arranged on the buckles 16, the buckles 16 can be preferably disassembled), the buckles 16 at the upper and lower sides outside the seeding depth quantifying barrel 1 loosen and pull the rope 18 to drive the pulleys 17 at the upper and lower opposite sides outside the substrate tray 3 (the pulleys 17 are preferably NSK pulleys) to move, so that the substrate tray 3 reaches the corresponding height, then, corresponding substrates (such as vermiculite, nutrient soil, field plowing soil, sand and the like) which are prepared in advance are loaded above the substrate tray 3 and the sowing depth quantifying barrel 1, then sowing seeds with corresponding quantity on the substrate, covering the substrate with corresponding thickness on the seeds, and finally numbering on the label groove 4 outside the sowing depth quantifying cylinder 1, and further carrying out the deep sowing germination test.
The foregoing is directed to embodiments of the present application and it is noted that numerous modifications and adaptations may be made by those skilled in the art without departing from the principles of the present application and are intended to be within the scope of the present application.

Claims (1)

1. A method for improving the deep sowing resistance of corn seeds is characterized by mainly comprising the following steps:
(1) seed selection: selecting the maize varieties K12 and Ji 853 which cannot be deeply sowed, and selecting the maize seeds with full, uniform and unbroken seeds from the maize varieties K12 and Ji 853;
(2) preparation of 24-Epinobrassinolide (EBR) solution: using 98% by volume of ethanol and ddH2O EBR was dissolved and diluted to 0, 1.0 and 2.0 mg L-1The EBR solution (1) of (1), the corresponding EBR solution finally contains 0.1% (v/v) of ethanol and 0.1% (v/v) of Tween-80;
(3) seed disinfection: firstly, disinfecting seeds for 10 min by using NaClO solution with the volume percentage of 0.5 percent, and then, using ddH2O washing the seeds for 3 times, and adsorbing water by using a sterilized filter paper;
(4) seed soaking: soaking the sterilized seeds in 0, 1.0 and 2.0 mg L at room temperature-1Soaking seeds in the EBR solution for 24 hours;
(5) seed of a plantPreparing a seed germination matrix: mixing sterilized vermiculite with 0, 1.0 and 2.0 mg. L-1The EBR solution is uniformly mixed with soil according to the proportion of 5g to 1mL to prepare a seed germination substrate;
(6) 15 and 20 cm seed depth sowing test: the above step (5) is carried out in an amount of 1.0 and 2.0 mg . L-1The seed germination substrate of EBR solution (2) was set in a seed deep-sowing test apparatus at a certain height, and then 1.0 and 2.0 mg were added . L-1The EBR solution is used for soaking 30 seeds, the seeds are respectively sown in the deep-sowing test device, and then the seeds are respectively covered by 15 cm and 20 cm to contain 1.0 mg and 2.0 mg . L-1The test of the seed germination substrate of the EBR solution of (1) is treated as an EBR solution deep-seeding; contains 0 mg of . L-1The seed germination matrix of the EBR solution is filled in a certain height of a seed deep-sowing test device, and then 0 mg of the seed germination matrix is added . L-1The EBR solution is used for soaking 30 seeds, sowing the seeds in a deep seed sowing test device, and covering the seeds by 3 cm, 15 cm and 20 cm respectively to contain 0 mg . L-1The test of the seed germination matrix of the EBR solution is used as positive normal control treatment CK (+), negative deep sowing stress control treatment CK (-)1 and negative deep sowing stress control treatment CK (-)2, and the test treatment is repeated for 3 times; after sowing, 0, 1.0 and 2.0 mg are respectively poured at intervals of 2 days . L-1The EBR solution (2) is 40 mL, and the solution is placed in the illumination for 12 hours per day with the illumination intensity of 600 mu mol/(s).m2) Germinating in a climatic chamber with day/night temperature of (25 + -1)/(20 + -1) deg.C and relative humidity of 60% for 10 d, and adding ddH2Quickly washing off vermiculite at the root of the seedling by using O water, adsorbing the attached water by using sterilizing filter paper, and measuring the corresponding deep-seeding resistance;
(7) evaluation of deep sowing effect of 15 and 20 cm seeds: measuring six phenotypic characters of emergence rate, mesocotyl length, embryo sheath length, sum of mesocotyl and embryo sheath, seedling length and root length under each treatment, respectively taking the hypocotyl and embryo sheath, and measuring H of corresponding mesocotyl and embryo sheath2O2Six physiological characters of content, lignin content, superoxide dismutase activity, peroxidase activity, catalase activity and ascorbic acid peroxidase activity; to evaluate the corresponding for convenienceThe deep sowing resistance of the corn with the exogenous EBR with the concentration under the condition of improving the deep sowing of the seeds of 15 cm and 20 cm, and the exogenous EBR deep sowing resistance adjustment index (RI) of the single trait of each measured trait is defined and calculated, and the formula is as follows:RI in =(T i-n -T CK i(-))/(|T CK(+)-T CK i(-)i) (a) orRI in =( T CK(-)i - T i-n )/(|T CK(+)-T CK i(-)L) (b), wherein:RI in is as followsiThe corresponding traits of the seeds under deep sowing stress are shown in the firstnExogenous EBR deep-seeding resistance regulation index, T, at seed concentrationCK(+)T is the measured value of the property under the positive normal control treatment CK (+)CK(-)iIs the negative direction
Figure DEST_PATH_IMAGE002
Trait measurement value, T, under control treatment of CK (-) i under seed deep-sowing stressi-nIs as follows
Figure 388311DEST_PATH_IMAGE002
First under the stress of deep sowing
Figure DEST_PATH_IMAGE004
Measuring the character value under the EBR solution with the seed concentration, if the measured character is in positive correlation with the deep sowing resistance of the corn, calculating by adopting the formula (a), otherwise, calculating by adopting the formula (b); the positive RI value indicates that the exogenous EBR with corresponding concentration plays a positive regulation role on the deep sowing stress, otherwise, the negative RI value indicates that the exogenous EBR with corresponding concentration plays a negative regulation role on the deep sowing stress, the exogenous EBR deep sowing resistance regulation index for measuring single character of each character is used as the evaluation index of the deep sowing resistance of the corn, and the membership function method is adopted to comprehensively evaluate the conditions of applying 1.0 mg and 2.0 mg under the deep sowing stresses of 15 cm and 20 cm . L-1The deep sowing resistance of the corn seeds after the exogenous EBR is strong and weak; the formula is as follows:U in =( RI in - RI imin )/(|RI imax -RI imin i), (c) orU ijn =1-(RI in - RI imin )/(|R imax -RI imin L) (d), wherein:U in is as followsiCorresponding traits under deep-sowing stress are imposednExogenous EBR deep-seeding resistance membership value of seed concentration,RI imin is as follows
Figure 721203DEST_PATH_IMAGE002
The corresponding traits of the seeds under deep sowing stress are shown in the firstnThe minimum RI value at seed concentration, and vice versa,RI imax is as followsiThe corresponding traits of the seeds under deep sowing stress are shown in the firstnMaximum RI at seed concentration; if the measured RI is positively correlated with the deep-seeding resistance regulation effect after the external source EBR is applied, calculating by adopting the formula (c), otherwise, calculating by adopting the formula (d), accumulating RI membership values of the external source EBR applied with various concentrations under the corresponding deep-seeding stress, and calculating the arithmetic mean of the RI membership values for comparison, wherein the larger the value is, the stronger the deep-seeding resistance of the corn after the external source EBR is applied is.
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Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63307803A (en) * 1987-06-09 1988-12-15 Sakata No Tane:Kk Production of brassicaceous crop seed
CN103238397A (en) * 2013-05-28 2013-08-14 甘肃农业大学 Screening method for deep-seeding-resisting corn germplasms
CN104313134A (en) * 2014-09-29 2015-01-28 甘肃农业大学 Method for researching deep seedling resistance of corn seeds based on microsatellite markers and subordination function method
CN104335734A (en) * 2014-11-18 2015-02-11 甘肃农业大学 Method for soaking seeds by epibrassinolide to relieve harm of different stresses on corn
CN104350834A (en) * 2014-10-09 2015-02-18 浙江农林大学 Seed soaking treatment method for increasing germination rate of corn seeds
CN207783900U (en) * 2018-01-15 2018-08-31 遵义医学院 A kind of orchid tissue culture device

Patent Citations (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPS63307803A (en) * 1987-06-09 1988-12-15 Sakata No Tane:Kk Production of brassicaceous crop seed
CN103238397A (en) * 2013-05-28 2013-08-14 甘肃农业大学 Screening method for deep-seeding-resisting corn germplasms
CN104313134A (en) * 2014-09-29 2015-01-28 甘肃农业大学 Method for researching deep seedling resistance of corn seeds based on microsatellite markers and subordination function method
CN104350834A (en) * 2014-10-09 2015-02-18 浙江农林大学 Seed soaking treatment method for increasing germination rate of corn seeds
CN104335734A (en) * 2014-11-18 2015-02-11 甘肃农业大学 Method for soaking seeds by epibrassinolide to relieve harm of different stresses on corn
CN207783900U (en) * 2018-01-15 2018-08-31 遵义医学院 A kind of orchid tissue culture device

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
不同玉米自交系耐深播性评价及遗传多样性分析;彭云玲等;《草业学报》;20160731;第25卷(第7期);第73-86页 *
不同玉米自交系耐深播能力的差异分析;彭云玲等;《干旱地区农业研究》;20140131;第32卷(第1期);第25-33,51页 *

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