CN115316267B - Olive germplasm aluminum resistance evaluation method and application thereof - Google Patents
Olive germplasm aluminum resistance evaluation method and application thereof Download PDFInfo
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Abstract
The invention belongs to the field of agricultural identification, and particularly relates to a large-scale evaluation method and application of olive seedling-stage aluminum resistance, and a system comprising the method. The method comprises the following steps: selecting olive seedlings as samples; placing the selected seedlings in an aluminum type Hoagland solution for stress simulated cultivation, and placing the samples in the Hoagland solution for simulated cultivation in a blank control group; determining tolerance indicators for the stress-simulated incubated samples compared to a blank control, the tolerance indicators comprising: the leaf fall rate, the rooting rate, the relative length of the prolonged root and the length of the new root; and calculating a factor score according to the tolerance index, accumulating the contribution rate, and grading the aluminum resistance of the sample according to an accumulated contribution rate grading rule and the factor score. The method is simple and efficient, can realize rapid in-vivo determination, effectively shortens the identification time, improves the efficiency, and provides powerful technical support for the identification and screening of the developed olive germplasm aluminum resistance germplasm.
Description
Technical Field
The invention belongs to the field of agricultural identification, and particularly relates to an olive germplasm aluminum resistance evaluation method and application thereof.
Background
Olea europaea is an evergreen small arbor of Olea of Oleaceae, and is one of four major oil plants in the world. Olive oil quality is reputed globally due to its abundance in unsaturated fatty acids and various antioxidant substances. Olives originate from subfamily asia and have been cultivated along the coast of the mediterranean for about 6000 years. In the beginning of the 20 th century, the olea europaea begins to spread rapidly to areas outside the Mediterranean sea, however, due to aluminum toxicity stress of acid red and yellow soil areas, the phenomena of blocked root system growth, serious leaf fall, weak growth vigor and the like of part of varieties occur, further development of the production of the olea europaea is seriously influenced, and no relevant data are recorded and reported about aluminum resistance of germplasm resources.
On the other hand, the production usually adopts a method of deep lime application to improve the soil pH value and reduce the aluminum toxicity, but the large-area lime application in mountainous and hilly areas greatly increases the planting cost and also destroys the soil physicochemical properties after long-term use. The method for directly identifying the field planting is very easily influenced by uncontrollable factors such as plant diseases and insect pests, soil uniformity and the like, and has the defects of large occupied area, high labor and time consumption and the like under the condition of more test materials, and is particularly not suitable for early large-scale identification of forest germplasm.
Therefore, the establishment of an olive germplasm aluminum resistance evaluation system and the development of germplasm early-stage large-scale aluminum resistance evaluation can provide an important method and basis for breeding new aluminum-resistant varieties and are also important preconditions for further popularization and application of the olives in the acid red-yellow soil region.
Disclosure of Invention
In order to solve the problem that no method capable of effectively evaluating the aluminum resistance of the olive germplasm on a large scale exists at present, the invention provides the method for evaluating the aluminum resistance of the olive germplasm and the application thereof, which can quickly and efficiently evaluate the aluminum resistance of the olive germplasm on a large scale, fill up the technical blank of evaluating the aluminum resistance of the olive germplasm, have important application value on screening the aluminum-resistant olive germplasm/variety and are beneficial to promoting the development of the olive industry in the acid red and yellow soil region.
The invention aims to:
1. the method can efficiently and quickly judge the aluminum resistance of the olive germplasm in the seedling stage;
2. the method ensures high judgment precision, has high confidence coefficient and can be effectively used for guiding production practice and the like.
In order to achieve the purpose, the invention adopts the following technical scheme.
A method for evaluating the aluminum resistance of olive germplasm,
the method comprises the following steps:
selecting olive seedlings as samples, wherein the seedling selection standard is as follows: the root system is 1-2 cm long, and the number of old leaves is 1-3; placing the selected seedlings in an aluminum type Hoagland solution for stress simulated cultivation, wherein the aluminum type Hoagland solution contains 40-60 mu mol/L of aluminum ions and is compared with a blank control group, and the blank control group places a sample in the Hoagland solution for simulated cultivation;
determining the tolerance index of the stress simulated culture sample compared with the blank control group after the simulated culture is carried out for at least 14 days, wherein the tolerance index comprises: leaf fall rate, rooting rate, root extension length and new root length;
and calculating a factor score of the selected tolerance index according to the tolerance index, calculating a cumulative contribution rate through the factor score, and evaluating the aluminum resistance of the sample according to a rating rule of the cumulative contribution rate and the factor score so as to grade the aluminum resistance of the olive germplasm.
As a preference, the first and second liquid crystal compositions are,
the factor score of the tolerance indicator is calculated by:
in the formula: fs i The score of the factor of the ith tolerance index, i =1,2,3,4, refers to the Defoliation Rate (DR), rooting Rate (RT), relative Length of Extended Root (LER) and Length of New Root (LNR), x ij The detection value of the ith tolerance index of the j sample for stress simulation cultivation is j =1,2 i0 The detection value of the ith tolerance index of the blank control group, x imax Simulation of incubation of samples for stress i An optimal value of the detection values of the individual tolerance indices;
calculating the score Gs of the common factor according to the factor scores i ，i＝1，2，3；
The common factor score Gs i The calculation method is as follows:
in the formula: gs 1 、Gs 2 And Gs 3 Respectively representing the scores of three common factors, fs 1 、Fs 2 、Fs 3 And Fs 4 Factor scores representing four tolerance indexes, a xy Factor variables each representing a factor score for the xth common factor score corresponding to the yth tolerance indicator, wherein a 11 ＝-0.1417，a 12 ＝0.1639，a 13 ＝0.9223，a 14 ＝0.6856，a 21 ＝0.9648，a 22 ＝-0.1164，a 23 ＝-0.0740，a 24 ＝-0.4098，a 31 ＝-0.1116，a 32 ＝0.9764，a 33 ＝0.1306，a 34 ＝0.2699；
Calculating the cumulative contribution rate W according to the public factor score i The process is as follows:
in the formula: c k Represents the contribution ratio of the kth common factor score, where C 1 ＝0.3418788，C 2 ＝0.2794388，C 3 ＝0.2639243，W i Represents the cumulative contribution of the ith commonality score, i =1,2,3.
As a preference, the first and second liquid crystal compositions are,
and the comprehensive aluminum resistance D value is calculated by identifying and evaluating the aluminum resistance of the sample according to the accumulated contribution rate rating rule and the factor score, and the comprehensive aluminum resistance D value is calculated by the following method:
in the formula: d represents the comprehensive aluminum resistance value, n =3,C i Represents the contribution of the ith common factor score, i =1,2,3, where C 1 ＝0.3418788，C 2 ＝0.2794388，C 3 ＝0.2639243，Gs i Score for the ith common factor, i =1,2,3, W 3 Representing the maximum cumulative contribution rate.
As a preference, the first and second liquid crystal compositions are,
the aluminum resistance is classified into five grades of high resistance, medium resistance, common, medium sensitivity and high sensitivity;
the D value range of the corresponding comprehensive aluminum resistance of the high resistance is 0.969-1.291;
the D value range of the middle-tolerance corresponding comprehensive aluminum tolerance is 0.259-0.828;
the general corresponding comprehensive aluminum resistance D value range is-0.191-0.209;
the D value range of the corresponding comprehensive aluminum resistance of the mesosensitive alloy is-0.726 to-0.247;
the D value range of the high-sensitivity corresponding comprehensive aluminum resistance is-1.699 to-0.839.
An application of an olive germplasm aluminum resistance evaluation method,
the method is used for olive germplasm resource screening.
The invention has the beneficial effects that:
through the analysis of the metadata, four basic tolerance indexes with highest confidence coefficient, lowest interference and highest efficiency are screened out from a plurality of factors, and the accuracy and the confidence coefficient of the identification and evaluation are greatly improved. Meanwhile, the experimental process is simple and efficient, rapid in vivo determination can be realized, the identification time is effectively shortened, the efficiency is improved, and powerful technical support is provided for developing the aluminum-resistant germplasm identification of the olive germplasm.
Drawings
FIG. 1 shows the results of a transplant test in an example of the present invention;
FIG. 2 is a graph comparing the relative length of the extended roots and the length of new roots after one and two weeks of treatment in the examples of the present invention.
Detailed Description
The invention is described in further detail below with reference to specific embodiments and drawings. Those skilled in the art will be able to practice the invention based on these descriptions. Furthermore, the embodiments of the present invention described in the following description are generally only a part of the embodiments of the present invention, and not all of the embodiments. Therefore, all other embodiments obtained by a person of ordinary skill in the art based on the embodiments of the present invention without any creative effort shall fall within the protection scope of the present invention.
Unless otherwise specified, the raw materials used in the examples of the present invention are all commercially available or available to those skilled in the art; unless otherwise specified, the methods used in the examples of the present invention are all those known to those skilled in the art.
Examples
The olive germplasm aluminum resistance identification evaluation test comprises:
the test was conducted in 2019 to 2021 at the academy of agricultural sciences in Zhejiang province, and a total of 97 olive germplasms (labeled #1 to # 97) were evaluated. All germplasms are bred by cutting in the last year by using perlite as a matrix, and are subjected to aluminum resistance identification treatment after about 4 months of cutting.
When the treatment is carried out: all cuttings grow 1.0-3.0 cm of new roots and contain 1-2 old leaves.
And (3) identifying the aluminum resistance of olive germplasm: taking 30 seedlings from 97 germplasm samples, wherein the seedling selection standard is as follows: the total length of a root system is 1-3 cm, the number of old leaves is 1-2, the residual matrix of the root is quickly washed by distilled water, the initial root length and the total number of leaves are measured, and then the root is placed in an aluminum type Hoagland solution for stress simulation cultivation, wherein the aluminum type Hoagland solution contains 50 mu mol/L of aluminum ions, and the pH is =5.0;
in the blank control group, a sample is placed in a Hoagland solution for simulated cultivation under the same conditions;
after the simulated cultivation is carried out for 14 days, the tolerance index of the stress simulated cultivation sample compared with the initial sample is determined, wherein after 50 germplasms are cultivated for 7 days, the related tolerance index is also determined.
Commonly tested indicators of tolerance include:
the change rate of the fresh weight/dry weight of the plant, the relative length of the branch, the change of the leaf area, the change of the stem thickness, the leaf fall rate, the rooting rate, the relative growth amount of the root system and the like;
the above are common tolerance indexes, but when plant tolerance is estimated, the most appropriate and relevant and significant indexes need to be screened from a plurality of tolerance indexes so as to improve the efficiency and accuracy of evaluation and identification and ensure higher confidence when the plant tolerance is used for guiding agriculture.
Through repeated observation and research, according to the cutting seedling raising mode of the olea europaea and the self growth and development characteristics, weak correlation tolerance indexes such as the change rate of fresh weight/dry weight, the length of branches, the change of leaf area/stem thickness and the like, and inconvenient rapid operation and stability indexes such as the activity of plant enzymes, photosynthesis or respiration change and the like are firstly eliminated, meanwhile, the root system is divided into two types of extended roots and new roots to be used as research, and finally four tolerance indexes with strong correlation with the aluminum resistance for the olea europaea are determined, namely the Defoliation Rate (DR), the rooting Rate (RT), the relative length of the extended roots (LER) and the new rooting Length (LNR), which are numbered as indexes from 1 to 4 in sequence.
The calculation is carried out by adopting a conventional method, and specifically comprises the following steps:
the formula for calculating the leaf fall rate index is as follows:
no factor is included;
the rooting rate index calculation formula is as follows:
no factor is included;
the relative length index calculation formula of the extension root is as follows: LER = total root length extension-initial length in cm;
the calculation formula of the new root length index is as follows: LNR = length of young root of new processes in cm.
The indexes correspond to calculation coefficients, and the marked index is numbered i, i =1,2,3,4, wherein the marked index is a deciduous leaf rate (DR) index when i =1, a rooting Rate (RT) index when i =2, an extended root relative Length (LER) index when i =3, and a new rooting Length (LNR) index when i =4. x is the number of ij The detection value of the ith tolerance index of the j test sample of the stress simulation breeding sample is j =1,2 101 That is, 1 st tolerance index [ Defoliation Rate (DR) index ] of sample No. 01]The detection value of (3).
Substituting the formula into the score calculation formula of the following factor to calculate:
in the formula: fs i The factor score for the ith tolerance index, i =1,2,3,4, sequentially refers to the Defoliation Rate (DR), rooting Rate (RT), root extension Length (LER) and new root Length (LNR), x, respectively ij The detection value of the ith tolerance index of the j sample for stress simulation cultivation is j =1,2 i0 The detection value of the ith tolerance index of the blank control group, x imax And (3) the optimal value of the detection value of the ith tolerance index of the stress simulation cultivation sample is not the maximum value, but the optimal value refers to the optimal performance, for example, the fallen leaf rate is lowest, the minimum value is taken as the optimal value, and the rooting rate, the root extension length and the new root length are all taken as the optimal values.
Calculating the score Gs of the common factor according to the factor scores i ，i＝1，2，3；
The common factor score Gs i The calculation method is as follows:
in the formula: gs 1 、Gs 2 And Gs 3 Respectively representing the scores of three common factors, fs 1 、Fs 2 、Fs 3 And Fs 4 Factor scores representing four tolerance indexes, a xy Factor variables representing the factor scores of the xth common factor score corresponding to the yth tolerance indicator, respectively.
In particular, the common factor score Gs used in the present invention i The calculation method is obtained by SPSS regression analysis calculation, and Correlation coefficient Matrix analysis of Correlation Matrix calculation, anti-image Correlation Matrix analysis of Anti-image Correlation Matrix calculation, bartlett test of sphere degree test analysis of probability and Kaiser-layer-Oklin Measure of sampling evaluation analysis are respectively carried out, and the results obtained by the accuracy are comprehensively tested and verified, the obtained common factor score calculation formula has significant influence on the results obtained by calculation, so that the accuracy of the results and the verification results reaches 100%, and the statistical difference P of each sample is less than 0.01 when the samples are examined, and the high confidence coefficient is achieved.
Calculating the cumulative contribution rate W according to the public factor score i The process is as follows:
in the formula:C k represents the contribution ratio of the kth common factor score, where C 1 ＝0.3418788，C 2 ＝0.2794388，C 3 ＝0.2639243，W i A cumulative contribution rate representing the ith common factor score, i =1,2,3;
calculating a comprehensive aluminum resistance D value based on the public factor score and the accumulated contribution rate;
the comprehensive aluminum resistance D value is calculated by the following method:
in the formula: d represents the comprehensive aluminum resistance value, n =3,C i Represents the contribution of the ith communal factor score, i =1,2,3, where C 1 ＝0，3418788，C 2 ＝0.2794388，C 3 ＝0.2639243，Gs i Score for the ith common factor, i =1,2,3, W 3 Representing the maximum cumulative contribution rate.
The aluminum resistance is classified into five grades of high resistance, medium resistance, general, medium sensitivity and high sensitivity;
the D value range of the corresponding comprehensive aluminum resistance of the high resistance is 0.969-1.291;
the D value range of the middle-tolerance corresponding comprehensive aluminum tolerance is 0.259-0.828;
the general corresponding comprehensive aluminum resistance D value range is-0.191-0.209;
the D value range of the corresponding comprehensive aluminum resistance of the mesosensitive alloy is-0.726 to-0.247;
the D value range of the high-sensitivity corresponding comprehensive aluminum resistance is-1.699 to-0.839.
The tolerance indexes of some samples (# 1 to #10 samples) during the recording experiment are shown in the following table. The data in the table are all the thirty test sample average values of each germplasm sample.
| x ij (i＝1，DR) | x ij (i＝2，RT) | x ij (i＝3，LER) | x ij (i＝4，LNR) | |
| #1 |
0 | 100.00％ | 7.10 | 4.79 |
| #2 sample | 3.33％ | 80.00％ | 7.57 | 4.35 |
| #3 sample | 4.04％ | 100.00％ | 8.00 | 3.27 |
| #4 sample | 9.70％ | 100.00％ | 6.90 | 3.86 |
| #5 sample | 10.00％ | 96.67％ | 9.28 | 2.67 |
| #6 sample | 20.56％ | 100.00％ | 12.73 | 5.69 |
| #7 sample | 26.67％ | 93.33％ | 8.89 | 4.21 |
| #8 sample | 5.00％ | 100.00％ | 6.58 | 4.03 |
| #9 sample | 9.68％ | 96.67％ | 7.78 | 4.40 |
| #10 sample | 22.22％ | 100.00％ | 6.72 | 2.33 |
Calculating and demonstrating based on the number of the #1 germplasm samples in the table;
placebo run data x in combination with #1 sample 10 ＝-6.67％，x 20 ＝100％，x 30 ＝7.17，x 40 =4.84 and records the data optimum value x of #1 sample 1max ＝0，x 2max ＝101％，x 3max ＝7.19，x 4max And 4.83, calculating to obtain a common factor characteristic value, a contribution rate and a cumulative contribution rate of each sample of the No. 1 germplasm samples.
| Common factor | Score value | Contribution rate C i (％) | Cumulative contribution rate W i (％) |
| Gs i ，i＝1 | -0.41984 | 34.18788 | 34.18788 |
| Gs i ，i＝2 | -1.2715 | 27.94388 | 62.13176 |
| Gs i ，i＝3 | 0.60296 | 26.39243 | 88.52419 |
And substituting the public factor score characteristic value, the contribution rate and the accumulated contribution rate to calculate the aluminum resistance D value of the #1 sample, wherein the calculation result is-0.383742, and three decimal places are taken, namely-0.384.
Sequentially inputting the x of the samples #1 to #97 by a system containing the method (recording all the algorithms in an Excel program of a computer and/or any conventional program software) ij Value data, x i0 Value data sum x imax In the value data, the aluminum resistance D values of all samples were calculated and the above-described classification was verified, and the results of all samples #1 to #97 are shown in the following table.
Randomly selecting 10 germplasms in each acid-resistant grade, and transplanting the germplasms into the aluminum-rich acid soil for field verification. After transplanting for 3 months, the survival rate of each germplasm is counted, and the relative growth amount of the plant height is measured. The results shown in FIG. 1 were obtained and the raw data were recorded as shown in the following table.
As can be seen from fig. 1 and the table above, the survival rate of the germplasm with high tolerance and medium tolerance is more than 90%, and the survival rates of the germplasm with general, medium sensitivity and high sensitivity are all lower than 90%; meanwhile, in each aluminum resistance grade, the relative growth quantity of the average plant height is gradually reduced along with the reduction of the aluminum resistance, and the result is consistent with the identification result.
In addition, comparative tests of the relative length of the elongation roots and the length of the new roots were performed on 50 of the germplasm aluminum type Hoagland solutions (50 μmol/L aluminum ion, pH = 5.0) treated for 7 days and 14 days. The results are shown in FIG. 2.
As can be seen from the figure, compared with 1 week of aluminum treatment, the tendency of various quality changes after 2 weeks of aluminum treatment is consistent, but the relative length of the prolonged roots and the length of the new roots among different germplasms are more different, and in order to reduce the test error, each tolerance index should be selected to be measured for about 2 weeks.
As a supplement, the method is verified by adopting a method of directly identifying field planting. Randomly selecting 10 germplasms from each aluminum-resistant grade, transplanting the germplasms into aluminum-rich acid soil, and verifying. After transplanting for 3 months, the survival rate of each germplasm is counted, and the relative growth amount of the plant height is measured.
As can be seen from figure 1, the survival rate of the germplasm with high tolerance and medium tolerance is more than 90 percent, and the survival rate of the germplasm with general tolerance, medium sensitivity and high sensitivity is less than 90 percent; meanwhile, in each aluminum resistance grade, the relative growth quantity of the average plant height is gradually reduced along with the reduction of the aluminum resistance, and is highly consistent with the result of the aluminum resistance identification in the invention.
In conclusion, the technical scheme of the invention can effectively and efficiently realize the identification and evaluation of the aluminum resistance of the olive germplasm, can realize rapid in vivo measurement, effectively shorten the identification time, improve the efficiency and provide powerful technical support for the identification and screening of the aluminum resistance of the olive germplasm.
Claims (3)
1. A method for evaluating the aluminum resistance of olive germplasm is characterized in that,
the method comprises the following steps:
selecting olive seedlings as samples, wherein the seedling selection standard is as follows: the root length is 1-2 cm, and the number of old leaves is 1-3;
placing the selected seedlings in an aluminum type Hoagland solution for stress simulated cultivation, wherein the aluminum type Hoagland solution contains 40-60 mu mol/L of aluminum ions and is compared with a blank control group, and the blank control group places a sample in the Hoagland solution for simulated cultivation;
after the simulated cultivation is carried out for at least 14d, determining the tolerance index of the stress simulated cultivation sample compared with a blank control group, wherein the tolerance index comprises: the leaf fall rate, the rooting rate, the relative length of the prolonged root and the length of the new root;
calculating a factor score of the selected tolerance index according to the tolerance index, calculating an accumulated contribution rate through the factor score, and identifying and evaluating the aluminum resistance of the sample according to an accumulated contribution rate rating rule and the factor score so as to realize the aluminum resistance rating of the olive germplasm;
the factor score of the tolerance indicator is calculated by:
in the formula:
is the first->
Factor scores for individual tolerance indicators>
In each case in turn denotes the falling leaf ratio (` H `)>
) (vii) rooting ratio (` Harbin `)>
) Extending the relative length of the root (` based `)>
) And new root length (` Harp `)>
)，/>
Incubation sample ^ th for stress simulation>
Test sample number ^ th>
A detection value for a respective tolerance indicator>
，/>
Is a blank control group->
A detection value of each tolerance index->
Incubating a sample for stress simulation ^ th>
An optimal value of the detection values of the individual tolerance indices;
calculating a common factor score based on the factor scores
，/>
；
The common factor score
The calculation method is as follows:
in the formula:
、/>
and &>
Represents three common factor scores, respectively>
、/>
、/>
And &>
Factor scores, representing four tolerance indices respectively, are evaluated>
Respectively denote a fifth->
Each common factor score corresponds to a ^ th ^ or ^ th>
A factor variable for a factor score of individual tolerance indicators, wherein->
=-0.1417，/>
=0.1639，/>
=0.9223，/>
=0.6856，/>
=0.9648，/>
=-0.1164，
=-0.0740，/>
=-0.4098，/>
=-0.1116，/>
=0.9764，/>
=0.1306，/>
=0.2699；
Calculating the accumulated contribution rate according to the public factor score
The process is as follows:
in the formula:
indicates the fifth->
A contribution rate of a common factor score, wherein
，/>
Represents a fifth or fifth party>
The cumulative contribution rate of the individual common factor scores,
；
and identifying and evaluating the aluminum resistance of the sample according to the accumulated contribution rate rating rule and the factor score to calculate a comprehensive aluminum resistance D value
The values are calculated by:
in the formula:
represents a comprehensive aluminum resistance value>
，/>
Indicates the fifth->
The contribution ratio of the individual common factor scores, <' >>
Wherein
，/>
Is a first->
The score of each of the common factors is, device for selecting or keeping>
，/>
Representing the maximum cumulative contribution rate.
2. The method of claim 1, wherein the method comprises the steps of,
the aluminum resistance is classified into five grades of high resistance, medium resistance, common, medium sensitivity and high sensitivity;
the high-tolerance comprehensive aluminum resistance
The value range is 0.969 to 1.291;
the medium-tolerance and corresponding comprehensive aluminum resistance
The value range is 0.259-0.828;
said general corresponding combined aluminium resistance
The value range is-0.191-0.209;
the mesosensitive corresponding comprehensive aluminum resistance
The value range is-0.726 to-0.247;
the high sensitivity corresponding comprehensive aluminum resistance
The value range is-1.699 to-0.839.
3. Use of the method according to claim 1 or 2,
the method is used for identifying olive germplasm resources.
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