CN105325192A - Identification method of salt-tolerant resource in seedling period of cowpea - Google Patents

Identification method of salt-tolerant resource in seedling period of cowpea Download PDF

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CN105325192A
CN105325192A CN201510783871.XA CN201510783871A CN105325192A CN 105325192 A CN105325192 A CN 105325192A CN 201510783871 A CN201510783871 A CN 201510783871A CN 105325192 A CN105325192 A CN 105325192A
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cowpea
salt
tolerant
seed
overall target
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张红梅
刘晓庆
陈华涛
陈新
崔晓艳
袁星星
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Jiangsu Academy of Agricultural Sciences
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Jiangsu Academy of Agricultural Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01G—HORTICULTURE; CULTIVATION OF VEGETABLES, FLOWERS, RICE, FRUIT, VINES, HOPS OR SEAWEED; FORESTRY; WATERING
    • A01G7/00—Botany in general
    • 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

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  • Environmental Sciences (AREA)
  • Biodiversity & Conservation Biology (AREA)
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  • Soil Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)

Abstract

The invention discloses an identification method of a salt-tolerant resource in the seedling period of cowpea, which is easy to operate and reliable in identification result. The identification method comprises the steps of carrying out germination experiment by adopting one cowpea variety to screen the optimal ultrasonic treatment method and determine the germination conditions; carrying out salt treatment experiment on three varieties by adopting salt solutions of multiple gradients to determine the optimal treatment condition; carrying out salt-tolerant experiment on 18 cowpea varieties of different genes, and carrying out principal component analysis to determine the optimal regression model. The identification method is used for identifying the salt-tolerant capability of the cowpea with 15d, and is short in identification time; the optimal salt-tolerant indexes of the cowpea are plant height, root length, plant dry weight and root dry weight, the identification method with simple operation and definite index is established, the shape indexes are visualized and convenient to count, and the standard identification method is provided for large-scale rapid identification of salt tolerance in the seedling period of the germplasm resource of the cowpea as well as the salt-tolerant variety selection and salt-tolerant mechanism.

Description

A kind of cowpea Salt Tolerance at Seedling Stage resource authentication method
Technical field
The invention belongs to eco-environment technology field, particularly relate to a kind of cowpea Salt Tolerance at Seedling Stage resource authentication method.
Background technology
According to UNESCO (UNESCO) and FAO (Food and Agriculture Organization of the United Nation) (FAO) incomplete statistics, whole world saline soil area about 1,000,000,000 hm 2(Wang Zunqin, 1993).China's saline soil area about 100,000,000 hm 2, these saline soil are parts of the important land resources of China, especially at northern area and the coastal south.The experimental results shows, the biological control measure of Screening to use salt-tolerant plant new varieties is one of the most economical effective method in improvement saline land (Shi Jinxia etc., 2004; Stapies, 1984).Because indoor qualification salt resistance has the advantages such as time short, capacity is large, repeatability is strong, environmental influence is little, to the indoor qualification in the extensive Salt-Tolerance Identification of crop many employings bud phase, seedling stage.The report of indoor Salt-Tolerance Identification method is a lot, and many researchers adopt different salt tolerant methods, filter out the germ plasm resource that a collection of salt resistance is strong, and starts to utilize (Wang Hongchun, 1989 in production; Chen Zhide etc., 2004].But work cowpea being carried out to germplasm identification evaluation seedling stage has no report.
Cowpea economic worth is higher, and its bean sprouts, seedling, tender pod all can eat as vegetables, is one of important vegetable solving the s supply of autumn in summer.China's Coastal Areas is the important locality of cowpea, and the salinity in the seawater of coastal area enters soil by ground and underground approach, and is in lower latitude, and the moisture evaporation of soil is more, also there is soil alkaline problem (Zhang Zhou etc., 2014).Therefore, Selection of Salt-Tolerant cowpea new varieties are one of most economical effective methods in improvement saline land.Utilize the controlled condition in greenhouse to develop water culture in recent years, the NaCl stress concentration adopted in different research is different, and authentication method is different, and identification of indicator is different, and analytical method is also different.Therefore, a set of cowpea Salt Tolerance at Seedling Stage resource authentication method being suitable for greenhouse controlled condition is needed.
Ultrasonic has promotion seed germination and root system development function, and using ultrasound ripple process seed has many research.Appoint (2010) such as Shekinahs to explore high-power ultrasonics to the impact of seed germination, show that ultrasonic 200W process 3 hours Germination of Soybean Seed rates are up to 97%.Chen Yanli etc. (2012) utilize ultrasonic process 25min wild soybean, improve the germination vigor of wild soybean, but do not have significant difference compared with the control.Li Gangs etc. (2007) utilize ultrasonic process Angelica Sinensis Seeds, can promote seed sprouting, accelerate growth of seedling, ahead of time crop maturity.Zhao faithful and upright person waits (2011) with excellent rice paddy seed for research object, have studied the influence of ultrasonic process to Germination Percentage of Rice and seedlings root.Result of study shows, ultrasonic Rice seeds treated can improve the growth of germination vigor and germination rate, promotion bud and root; Frequency is 25kHz, and the processing time is that the effect of 20min is better.These results of study illustrate that ultrasonic process may increase the permeability of crop seed coat, are conducive to seed sprouting.But the impact of ultrasonic process on cowpea seed germination rate does not also have relevant report.
In recent years, to the research of plant salt endurance there are reports (Tuncturketal., 2008; Yuetal., 2001; Baietal., 2009).Parker etc. (1987) study discovery, and the effect of salt pair soy bean toxic evil, causes plant leaf wither and come off.Shao Guihua etc. (1993) assessment method to soybean salt-tolerance is studied, and identifies the salt resistance of 1716 parts of soybean germplasms.Authentication method a lot (Wei Xiuying etc., 2008 of soybean varieties salt resistance; Wang Min etc., 2005; Qiao Yake etc., 2002; When horse is refined etc., 1994; Huetal., 1997; Kou He etc., 2007), as Sha Peifa (Wei Xiuying etc., 2008), nutrient solution screening method (Wang Min etc., 2005) etc.But authentication method is complicated mostly, and determination period is long.About the research report of cowpea salt resistance is less, Zu Yanxia etc. (2014) salting liquid of 9 ‰ to have carried out Salt-Tolerance Identification and the screening of bud phase to 17 parts of Cowpea Germplasm Resources, filter out one 3 grades salt tolerant cowpea variety Long Xiangte kings' morning.Zu Yanxia etc. (2012), with analyzing pure NaCl, prepare the salting liquid of 0.3%, 0.6%, 09% and 1.2% respectively, have studied the impact of salt stress on purple pod and green pod cowpea variety seed germination.Result shows, the relative salt damage rate of cowpea seed and salt tolerant rank increase along with salinity and raise.Size (the Sun little Fang etc. of screening concentration meeting influence index difference between screening material, 2001), suitable screening pressure energy enough makes index of correlation become large (peace Yongping etc., 2006) at the variation amplitude of storeroom and the coefficient of variation, obtains good screening effect.Research cowpea being carried out to germplasm identification evaluation seedling stage has no report.Therefore, screening is suitable for cowpea to carry out the salt stress concentration of germplasm identification seedling stage most important.
Current numerous salt tolerance of crop qualification adopts the overall merits such as multiple phenotypic character index, physiological and biochemical index, just has nothing in common with each other in analytical method.From early stage variance analysis and hierarchical clustering method (Zhang Jin is big, 2004) principal component analysis (PCA) (Xie Songfeng etc., 2010) finally and subordinate functional analysis method (Zhang Guoxin etc., 2011).Liu Yahui etc. (2012) comprehensively above-mentioned 3 kinds of methods utilize the method overall merit upland cotton Seedling Salt-tolerance such as Grey Correlation Cluster, principal component analysis and successive Regression, and it is that the result degree of correlation of identification of indicator evaluation reaches 0.990 that the physical signs filtered out carries out the result of salt resistance overall merit and salt gypsum rock.This equation not only can distinguish salt tolerant and salt density value material exactly, can carry out quantitatively to material salt tolerant (salt density value) degree according to y value and our standard of setting simultaneously, if for salt pond, to be conducive to identifying colony, improve the accuracy of QTL location or association analysis.The mensuration of selected index is quick and convenient, does not hinder plant, also can be improved as required, thus be applied to more wide field.
Summary of the invention
The object of the present invention is to provide a kind of cowpea Salt Tolerance at Seedling Stage resource authentication method, be intended to set up one simple to operate, the authentication method that index is clear and definite, selects for the Large scale identification of Cowpea Germplasm Resources Seedling Salt-tolerance and salt-enduring cultivars and Mechanisms of Salt Resistance provides the authentication method of standard.
The present invention is achieved in that a kind of cowpea Salt Tolerance at Seedling Stage resource authentication method, and described cowpea Salt Tolerance at Seedling Stage resource authentication method comprises the following steps:
Small-power ultrasonic process cowpea seed: to sterilize 10min with 1% hypochlorite solution, then use distilled water flushing 5 times, employing power output is 50W, and frequency is 40kHZ, and Germination Temperature is 25 DEG C, and the processing time is 3h, promotes Seed imbibition and sprouting;
Sowing: the cowpea seed of 18 ultrasonic process is seeded in the plastic cup filling sterilizing vermiculite, broadcasts 6 for every glass, then cover 2cm vermiculite, process and contrast are placed in a plastic casing respectively;
Thinning: thinning after true leaf launches completely, every glass is stayed 4 strains;
Ficus caricaL: with 400mmolL -1the 1/2Hoagland culture fluid process of NaCl, simultaneously not add the 1/2Hoagland nutrient solution of NaCl for contrast, changed a salting liquid every 3 days;
Copy kind: until salt density value material is dead, measure the salt tolerant rate of process and check sample, chlorophyll content, plant height, root length, plant fresh weight, root fresh weight, plant weights and root dry weight;
Data statistics: data preparation and analysis Bian MicrosoftExcel2010 software; SAS9.0 software is adopted to carry out the multivariate analysis of principal component analysis, cluster analysis and successive Regression.
Further, described cowpea seed to be sterilized 10min with 1% hypochlorite solution, then uses distilled water flushing 5 times, the little yarn bag being placed with 200 seeds is immersed in below the excusing from death washer water surface, water temperature is 30 DEG C, hyperacoustic power output is 50W, and frequency is 40kHZ, and the processing time is 3h, every 50 1 group, the seed handled well is placed in 12cm culture dish to be sprouted, repeat for 4 times, two metafiltration paper are seedbed, add sterile water and keep seedbed moistening; Seed is placed in illumination box and cultivates, and Germination Temperature is 25 DEG C, and start record after 24h and sprout number, every 24h records once, with the experimental group without ultrasonic process for control group.
Further, described NaCl Ficus caricaL comprises: select 18 cowpea varieties (varietal characteristic: sowing is to the tender pod 55-65 days that gathers, picking time 25-35 days, the time of infertility 85-90 days), 18 cowpea variety seeds to be placed in cloth bag airing 1 day, and employing power output is 50W, and frequency is that planting seed after the process of 40kHZ ultrasonic is in the plastic cup filling sterilizing vermiculite, every basin broadcasts 6, then covers 2cm vermiculite.
Further, described data statistics adopts the salt tolerant efficiency of different single index to evaluate cowpea salt resistance, specifically comprises:
Each test index salt tolerant efficiency STC:
STC=process measured value/blank determination value × 100%;
The membership function value of each overall target of different genotype cowpea:
u(X j)＝(X j-X min)/(X max-X min)j＝1,2,…,n；
In formula, X jrepresent a jth overall target; X minrepresent the minimum of a value of a jth overall target; X maxrepresent the maximum of a jth overall target;
The weight of each overall target:
w j = p j / Σ j = 1 n p j , j = 1 , 2 , ... , n ;
In formula, w jrepresent significance level in all overall targets of a jth overall target and weight; p jrepresent the contribution rate through each cowpea variety jth overall target of principal component analysis gained;
The size of the comprehensive salt resistance ability of each cowpea variety:
D = Σ j = 1 n [ u ( X j ) × w j ] , j = 1 , 2 , ... , n ;
In formula, D value is each cowpea variety is evaluated gained under condition of salt stress salt resistance comprehensive evaluation value by overall target.
Further, described data statistics utilizes successive Regression to establish cowpea Seedling Salt-tolerance and evaluates regression model D=-0.058+0.002PH+0.003RL+0.002PDW+0.002RDW (R 2=0.9797) 4 the single index plant heights, root length, overground part dry weight and the root dry weight that significantly affect cowpea seedling salt resistance ability, is filtered out.
Cowpea Salt Tolerance at Seedling Stage resource authentication method provided by the invention, employing vermiculite is matrix; Adopt 100,200,300 and 400mmol ﹒ L – 1naCl concentration process 5 cowpea varieties carry out preliminary experiment, determine 400mmol ﹒ L – 1naCl concentration is best screening concentration; Adopt the method for Nacl solution immersion treatment; The salt resistance ability of cowpea is identified in process over 15 days, qualification time is shorter; Determining the best salt-tolerance index of cowpea is plant height, root length, plant weights and root dry weight.
Accompanying drawing explanation
Fig. 1 is the cowpea Salt Tolerance at Seedling Stage resource authentication method flow chart that the embodiment of the present invention provides.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with embodiment, the present invention is further elaborated.Should be appreciated that specific embodiment described herein only in order to explain the present invention, be not intended to limit the present invention.
It is matrix with vermiculite that the present invention establishes a kind of, uses 400mmolL – 1naCl process, 15d can carry out the short-cut method of cowpea Seedling Salt-tolerance qualification, for the Large scale identification of Cowpea Germplasm Resources Seedling Salt-tolerance and the research of salt-enduring cultivars selection and Mechanisms of Salt Resistance provide new method.
Below in conjunction with accompanying drawing, application principle of the present invention is explained in detail.
As shown in Figure 1, the cowpea Salt Tolerance at Seedling Stage resource authentication method of the embodiment of the present invention comprises the following steps:
S101: small-power ultrasonic process cowpea seed: to sterilize 10min with 1% hypochlorite solution, then use distilled water flushing 5 times, employing power output is 50W, and frequency is 40kHZ, and Germination Temperature is 25 DEG C, and the processing time is 3h, promotes Seed imbibition and sprouting;
S102: sowing: the cowpea seed of 18 ultrasonic process is seeded in the plastic cup filling sterilizing vermiculite, broadcasts 6 for every glass, then cover 2cm vermiculite, process and contrast are placed in a plastic casing respectively;
S103: thinning: thinning after true leaf launches completely, every glass is stayed 4 strains;
S104: Ficus caricaL: with 400mmolL -1the 1/2Hoagland culture fluid process of NaCl, simultaneously not add the 1/2Hoagland nutrient solution of NaCl for contrast, changed a salting liquid every 3 days;
S105: copy kind: until salt density value material is dead, measures the salt tolerant rate of process and check sample, chlorophyll content, plant height, root length, plant fresh weight, root fresh weight, plant weights and root dry weight;
S106: data statistics: data preparation and analysis Bian MicrosoftExcel2010 software; SAS9.0 software is adopted to carry out the multivariate analysis of principal component analysis, cluster analysis and successive Regression.
In conjunction with following experiment, application principle of the present invention is further described.
1) seed treatment: cowpea variety is Soviet Union's No. 1, cowpea, medium variety, resistance to Cowpea viral disease.Plant height about 3.5 meters, the long 70-75cm of pod, thick 0.9cm, single pod weighs 27.32 grams.Sow to gathering tender pod about 55 days, picking time 25-30 days, the cowpea variety in the 88 days time of infertility.After choosing seed, to place it in cloth bag airing 1 day, then adopt the process of small-power ultrasonic.
Preliminary experiment processing procedure is as follows: cowpea seed to be sterilized 10min with 1% hypochlorite solution, then uses distilled water flushing 5 times.The little yarn bag being placed with 200 seeds is immersed in below the excusing from death washer water surface, water temperature is 30 DEG C, and hyperacoustic power output is 50W, and frequency is 40kHZ, and the processing time is 1h, 2h, 3h, 4h, 5h, 6h.Every 50 1 group, the seed handled well is placed in 12cm culture dish to be sprouted, and repeat for 4 times, two metafiltration paper are seedbed, adds sterile water and keeps seedbed moistening.Seed is placed in illumination box and cultivates, and Germination Temperature is 25 DEG C, and start record after 24h and sprout number, every 24h records once, with the experimental group without ultrasonic process for control group.Empirical average result is as follows:
Processing time (hour) Final germination rate (%) With the difference contrasted Effect
1h 71 Significantly Promote
2h 81 Significantly Promote
3h 96 Extremely remarkable Promote
4h 87 Significantly Promote
5h 89 Significantly Promote
6h 84 Significantly Promote
According to above experimental result, determine that cowpea optimal treatment condition be water temperature is 30 DEG C, hyperacoustic power output is 50W, and frequency is 40kHZ, and Germination Temperature is 25 DEG C, and the processing time is 3h.After seed sprouting (radicle length >=1cm), be placed on after wrapping with wet gauze in vacuum flask, keep temperature 24 ~ 30 DEG C, treat that sowing uses.
2) Nacl Ficus caricaL: cowpea variety is No. 1, cowpea morning, Soviet Union's No. 1, cowpea, Soviet Union's No. 3, cowpea, all resistance to Cowpea viral disease.About plant height 3.0-3.5 rice, the long 55-75cm of pod, the heavy 22-30 gram of single pod.Sow to tender pod of gathering and be about 55-65 days, picking time 25-35 days, the cowpea variety in the 85-90 days time of infertility.After choosing seed, to place it in cloth bag airing 1 day, adopt above-mentioned germination condition, by the planting seed after process in the plastic cup filling sterilizing vermiculite, every basin broadcasts 6, then covers 2cm vermiculite.
Preliminary experiment processing procedure is as follows:
Preparation pancebrin: KNO 36.5mM; Ca (NO 3) 2.4H 2o4.0mM; NH 4h 2pO 42mM; MgSO 4.7H 2o2.0mM; H 3bO 34.6mM; MnCl 2.4H 2o0.5 μM; ZnSO 4.7H 2o0.2 μM; (NH 4) 6mo 7o 24.4H 2o0.1 μM; CuSO 4.5H 2o0.2 μM; FeCl 345 μMs.
Thinning after 3 cowpea variety true leaves launch completely, every glass is stayed 4 strains.Adopt 100,200,300,400 and 500mmolL -1the 1/2Hoagland culture fluid process of NaCl concentration, compares with the 1/2Hoagland nutrient solution not adding NaCl simultaneously.Changed a salting liquid every 3 days, until salt density value material death (processing latter 15 days), range estimation salt tolerant rate (STR), salt tolerant rate is divided into 5 grades, from 1 grade (death) to 5 grades (normally).Chlorophyll content (SPAD value) is measured with chlorophyll meter (KonicaMinoltaSPAD-502).Empirical average result is as follows:
Note: in table, data are repeat mean value 3 times, and letter is for utilizing sas8.01 to carry out the significance of difference analyzed.Lowercase alphabet differential is different reaches 0.05 significance level, and capitalization represents that difference reaches 0.01 significance level.
According to above experimental result, adopt the process of different N aCl concentration after 15 days, the STR value of control group is 5.0, along with the raising (100-500mmolL of concentration NaCl concentration in processed group -1), 3 cowpea varieties show different salt tolerant degree, and No. 1, cowpea performance of wherein reviving is extremely resistance to, early cowpea 1 number it, Soviet Union's cowpea No. 3 sensitivities.Along with the increase of Nacl concentration, sensitive varieties Soviet Union's cowpea No. 3 SPAD values show as the trend of reduction, when concentration is increased to 500mmolL -1time, SPAD value is minimum, and STR value is also minimum, and plant performance is dead; Salt-enduring cultivars Soviet Union's cowpea No. 1 SPAD value shows as the trend first increasing and reduce afterwards, when concentration is increased to 400mmolL from 100 -1time, SPAD value is the highest, and STR value is 4.5, and plant is acted normally, when concentration is increased to 500mmolL -1time, SPAD value and STR value reduce, plant performance yellow.
According to above experimental result, determine that cowpea salt tolerant optimum condition is 400mmolL -1the 1/2Hoagland culture fluid process of NaCl 15 days, range estimation salt tolerant rate (STR) and measurement chlorophyll content (SPAD value).
3) screening of cowpea Salinity tolerance germplasm resource
1, small-power ultrasonic process cowpea seed: to sterilize 10min with 1% hypochlorite solution, then use distilled water flushing 5 times.Employing power output is 50W, and frequency is 40kHZ, and Germination Temperature is 25 DEG C, and the processing time is 3h, promotes Seed imbibition and sprouting;
2, sow: the cowpea seed of 18 ultrasonic process is seeded in the plastic cup filling sterilizing vermiculite, broadcasts 6 for every glass, then cover 2cm vermiculite.Process and contrast are placed in a plastic casing respectively;
3, thinning: thinning after true leaf launches completely, every glass is stayed 4 strains;
4, Ficus caricaL: with 400mmolL -1the 1/2Hoagland culture fluid process of NaCl, simultaneously not add the 1/2Hoagland nutrient solution of NaCl for contrast, changed a salting liquid every 3 days;
5, copy kind: until salt density value material death (processing latter 15 days), measure the salt tolerant rate (STR) of process and check sample, chlorophyll content (SPAD), plant height (PH), root long (RH), plant fresh weight (PFW), root fresh weight (RFW), plant weights (PDW) and root dry weight (RDW).
6, data statistics: data preparation and analysis Bian MicrosoftExcel2010 software; SAS9.0 software is adopted to carry out the multivariate analyses such as principal component analysis, cluster analysis and successive Regression.
Because the amplitude of variation of each single index of different cowpea materials is different, therefore adopt the salt tolerant efficiency of different single index to evaluate cowpea salt resistance.
(1) each test index salt tolerant efficiency (Salttolerantcoefficient, STC)
STC=process measured value/blank determination value × 100% 1.
(2) membership function value of each overall target of different genotype cowpea
u(X j)＝(X j-X min)/(X max-X min)j＝1,2,…,n②
In formula, X jrepresent a jth overall target; X minrepresent the minimum of a value of a jth overall target; X maxrepresent the maximum of a jth overall target.
(3) weight of each overall target
w j = p j / Σ j = 1 n p j , j = 1 , 2 , ... , n ③
In formula, w jrepresent significance level in all overall targets of a jth overall target and weight; p jrepresent the contribution rate through each cowpea variety jth overall target of principal component analysis gained.
(4) size of the comprehensive salt resistance ability of each cowpea variety
D = Σ j = 1 n [ u ( X j ) × w j ] , j = 1 , 2 , ... , n ④
In formula, D value is each cowpea variety is evaluated gained under condition of salt stress salt resistance comprehensive evaluation value by overall target.
7, result:
(1) principal component analysis: convert cowpea 8 single indexs in seedling stage under salt stress process to 4 overall targets, research establishes the regression model of cowpea plant Evaluation of Salt Tolerance, and obtain comprehensive evaluation value D, reflect the salt resistance of each tested variety more objectively.
(2) cluster analysis: 18 cowpea varieties are divided into salt density value type (6 parts), weak salt-resistant type (4 parts), moderate salt-resistant type (4 parts) and height salt-resistant type (4 parts).
(3) successive Regression: utilize successive Regression to establish cowpea Seedling Salt-tolerance and evaluate regression model D=-0.058+0.002PH+0.003RL+0.002PDW+0.002RDW (R 2=0.9797), 4 single index plant heights (PH), root that filtering out significantly affect cowpea seedling salt resistance ability grow (RL), overground part dry weight (PDW) and root dry weight (RDW), and the estimated accuracy of each material is all more than 99.1%, disclose the indexs such as PH, RL, PDW and RDW on the impact of cowpea salt resistance obviously, can be used for cowpea Evaluation of Salt Tolerance.
The foregoing is only preferred embodiment of the present invention, not in order to limit the present invention, all any amendments done within the spirit and principles in the present invention, equivalent replacement and improvement etc., all should be included within protection scope of the present invention.

Claims (5)

1. a cowpea Salt Tolerance at Seedling Stage resource authentication method, is characterized in that, described cowpea Salt Tolerance at Seedling Stage resource authentication method comprises the following steps:
Small-power ultrasonic process cowpea seed: to sterilize 10min with 1% hypochlorite solution, then use distilled water flushing 5 times, employing power output is 50W, and frequency is 40kHZ, and Germination Temperature is 25 DEG C, and the processing time is 3h, promotes Seed imbibition and sprouting;
Sowing: the cowpea seed of 18 ultrasonic process is seeded in the plastic cup filling sterilizing vermiculite, broadcasts 6 for every glass, then cover 2cm vermiculite, process and contrast are placed in a plastic casing respectively;
Thinning: thinning after true leaf launches completely, every glass is stayed 4 strains;
Ficus caricaL: with 400mmolL -1the 1/2Hoagland culture fluid process of NaCl, simultaneously not add the 1/2Hoagland nutrient solution of NaCl for contrast, changed a salting liquid every 3 days;
Copy kind: until salt density value material is dead, measure the salt tolerant rate of process and check sample, chlorophyll content, plant height, root length, plant fresh weight, root fresh weight, plant weights and root dry weight;
Data statistics: data preparation and analysis Bian MicrosoftExcel2010 software; SAS9.0 software is adopted to carry out the multivariate analysis of principal component analysis, cluster analysis and successive Regression.
2. cowpea Salt Tolerance at Seedling Stage resource authentication method as claimed in claim 1, it is characterized in that, described cowpea seed to be sterilized 10min with 1% hypochlorite solution, use distilled water flushing again 5 times, the little yarn bag being placed with 200 seeds is immersed in below the excusing from death washer water surface, water temperature is 30 DEG C, hyperacoustic power output is 50W, frequency is 40kHZ, processing time is 3h, and every 50 1 group, the seed handled well is placed in 12cm culture dish to be sprouted, and repeats for 4 times, two metafiltration paper are seedbed, add sterile water and keep seedbed moistening; Seed is placed in illumination box and cultivates, and Germination Temperature is 25 DEG C, and start record after 24h and sprout number, every 24h records once, with the experimental group without ultrasonic process for control group.
3. cowpea Salt Tolerance at Seedling Stage resource authentication method as claimed in claim 1, it is characterized in that, described NaCl Ficus caricaL comprises: select 18 cowpea varieties, 18 cowpea variety seeds to be placed in cloth bag airing 1 day, employing power output is 50W, frequency be planting seed after the process of 40kHZ ultrasonic in the plastic cup filling sterilizing vermiculite, every basin broadcasts 6, then covers 2cm vermiculite; 18 cowpea variety features: sowing to the tender pod 55-65 days that gathers, picking time 25-35 days, the time of infertility 85-90 days.
4. cowpea Salt Tolerance at Seedling Stage resource authentication method as claimed in claim 1, it is characterized in that, described data statistics adopts the salt tolerant efficiency of different single index to evaluate cowpea salt resistance, specifically comprises:
Each test index salt tolerant efficiency STC:
STC=process measured value/blank determination value × 100%;
The membership function value of each overall target of different genotype cowpea:
u(X j)＝(X j-X min)/(X max-X min)j＝1,2,...,n；
In formula, X jrepresent a jth overall target; X minrepresent the minimum of a value of a jth overall target; X maxrepresent the maximum of a jth overall target;
The weight of each overall target:
w j = p j / Σ j = 1 n p j , j = 1 , 2 , ... , n ;
In formula, w jrepresent significance level in all overall targets of a jth overall target and weight; p jrepresent the contribution rate through each cowpea variety jth overall target of principal component analysis gained;
The size of the comprehensive salt resistance ability of each cowpea variety:
D = Σ j = 1 n [ u ( X j ) × w j ] , j = 1 , 2 , ... , n ;
In formula, D value is each cowpea variety is evaluated gained under condition of salt stress salt resistance comprehensive evaluation value by overall target.
5. cowpea Salt Tolerance at Seedling Stage resource authentication method as claimed in claim 1, it is characterized in that, described data statistics utilizes successive Regression to establish cowpea Seedling Salt-tolerance and evaluates regression model D=-0.058+0.002PH+0.003RL+0.002PDW+0.002RDW (R 2=0.9797) 4 the single index plant heights, root length, overground part dry weight and the root dry weight that significantly affect cowpea seedling salt resistance ability, is filtered out.
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CN105830741A (en) * 2016-06-08 2016-08-10 江苏省农业科学院 Medium maturity cowpea variety outdoor spring sowing planting method
CN106069696A (en) * 2016-07-07 2016-11-09 中国农业科学院棉花研究所 Semi-wild is cotton to water planting Resistance Identification method in secondary saline and alkaline seedling stage
CN107853054A (en) * 2017-11-22 2018-03-30 中国热带农业科学院椰子研究所 Coconut Evaluation of Cold Tolerance method based on mathematical model
CN108207170A (en) * 2018-01-31 2018-06-29 江苏沿海地区农业科学研究所 A kind of integrated evaluating method of garlic sprouting stage salt tolerance
CN108241043A (en) * 2018-01-22 2018-07-03 山东省农业科学院作物研究所 A kind of identification method of sweet potato Seedling Salt-tolerance
CN108315465A (en) * 2018-03-27 2018-07-24 江苏省农业科学院 A kind of and cowpea salt tolerant correlated traits close linkage InDel molecular labelings and its primer and application
CN108377788A (en) * 2018-05-31 2018-08-10 中国农业科学院作物科学研究所 A kind of rice at whole growth periods Salt-Tolerance Identification evaluation method
CN108739196A (en) * 2018-05-18 2018-11-06 江苏省农业科学院 A kind of soybean salt-tolerance composite sifting and identification method
CN108776821A (en) * 2018-06-15 2018-11-09 河北省农林科学院经济作物研究所 A method of screening degeneration-resistant resource using plant phenotypic characteristics
CN108950054A (en) * 2018-08-28 2018-12-07 江苏省农业科学院 A kind of InDel molecular marker and primer thereof and application with cowpea salt tolerant correlated traits close linkage
CN109187863A (en) * 2018-09-03 2019-01-11 河北省农林科学院滨海农业研究所 A kind of greening seedling Seedling Salt-tolerance identification method
CN109565999A (en) * 2019-01-17 2019-04-05 甘肃省农业科学院作物研究所 A kind of flax kind germination period salt tolerance rapid identification method
CN110199711A (en) * 2019-06-20 2019-09-06 黑龙江省农业科学院耕作栽培研究所 A kind of mould accurate identification method of Resistance To Root Rot Disease resource of big bean curd
CN111855751A (en) * 2020-08-14 2020-10-30 南通大学 Method for rapidly evaluating salt tolerance of willow variety based on resistance value
CN112273087A (en) * 2020-10-30 2021-01-29 黑龙江八一农垦大学 Method for identifying saline-alkali resistance of soybeans
CN115280931A (en) * 2022-09-02 2022-11-04 华中农业大学 Seed treatment method for improving stain resistance

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CN105830741A (en) * 2016-06-08 2016-08-10 江苏省农业科学院 Medium maturity cowpea variety outdoor spring sowing planting method
CN106069696B (en) * 2016-07-07 2019-05-17 中国农业科学院棉花研究所 Semi-wild cotton is to secondary saline and alkaline seedling stage water planting Resistance Identification method
CN106069696A (en) * 2016-07-07 2016-11-09 中国农业科学院棉花研究所 Semi-wild is cotton to water planting Resistance Identification method in secondary saline and alkaline seedling stage
CN107853054A (en) * 2017-11-22 2018-03-30 中国热带农业科学院椰子研究所 Coconut Evaluation of Cold Tolerance method based on mathematical model
CN108241043A (en) * 2018-01-22 2018-07-03 山东省农业科学院作物研究所 A kind of identification method of sweet potato Seedling Salt-tolerance
CN108207170A (en) * 2018-01-31 2018-06-29 江苏沿海地区农业科学研究所 A kind of integrated evaluating method of garlic sprouting stage salt tolerance
CN108315465A (en) * 2018-03-27 2018-07-24 江苏省农业科学院 A kind of and cowpea salt tolerant correlated traits close linkage InDel molecular labelings and its primer and application
CN108315465B (en) * 2018-03-27 2021-07-06 江苏省农业科学院 InDel molecular marker closely linked with cowpea salt tolerance related characters, primers and application thereof
CN108739196B (en) * 2018-05-18 2020-09-04 江苏省农业科学院 Salt tolerance compound screening and identifying method for soybeans
CN108739196A (en) * 2018-05-18 2018-11-06 江苏省农业科学院 A kind of soybean salt-tolerance composite sifting and identification method
CN108377788A (en) * 2018-05-31 2018-08-10 中国农业科学院作物科学研究所 A kind of rice at whole growth periods Salt-Tolerance Identification evaluation method
CN108776821A (en) * 2018-06-15 2018-11-09 河北省农林科学院经济作物研究所 A method of screening degeneration-resistant resource using plant phenotypic characteristics
CN108776821B (en) * 2018-06-15 2020-07-28 河北省农林科学院经济作物研究所 Method for screening stress-resistant resources by using plant phenotypic characters
CN108950054A (en) * 2018-08-28 2018-12-07 江苏省农业科学院 A kind of InDel molecular marker and primer thereof and application with cowpea salt tolerant correlated traits close linkage
CN109187863A (en) * 2018-09-03 2019-01-11 河北省农林科学院滨海农业研究所 A kind of greening seedling Seedling Salt-tolerance identification method
CN109565999A (en) * 2019-01-17 2019-04-05 甘肃省农业科学院作物研究所 A kind of flax kind germination period salt tolerance rapid identification method
CN109565999B (en) * 2019-01-17 2021-11-05 甘肃省农业科学院作物研究所 Rapid identification method for salt tolerance of flax variety in germination period
CN110199711A (en) * 2019-06-20 2019-09-06 黑龙江省农业科学院耕作栽培研究所 A kind of mould accurate identification method of Resistance To Root Rot Disease resource of big bean curd
CN111855751A (en) * 2020-08-14 2020-10-30 南通大学 Method for rapidly evaluating salt tolerance of willow variety based on resistance value
CN111855751B (en) * 2020-08-14 2021-02-19 南通大学 Method for rapidly evaluating salt tolerance of willow variety based on resistance value
CN112273087A (en) * 2020-10-30 2021-01-29 黑龙江八一农垦大学 Method for identifying saline-alkali resistance of soybeans
CN115280931A (en) * 2022-09-02 2022-11-04 华中农业大学 Seed treatment method for improving stain resistance

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