CN105359856A - Method and device for batch identification of wheat salt tolerance - Google Patents
Method and device for batch identification of wheat salt tolerance Download PDFInfo
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- CN105359856A CN105359856A CN201510766259.1A CN201510766259A CN105359856A CN 105359856 A CN105359856 A CN 105359856A CN 201510766259 A CN201510766259 A CN 201510766259A CN 105359856 A CN105359856 A CN 105359856A
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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
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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
- A01G9/00—Cultivation in receptacles, forcing-frames or greenhouses; Edging for beds, lawn or the like
- A01G9/02—Receptacles, e.g. flower-pots or boxes; Glasses for cultivating flowers
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Abstract
The invention discloses a method and device for identification of plant salt tolerance. The device comprises a box body of which the upper end is open, wherein a plurality of pipelines are disposed in the box body, the positions of the pipelines are adjustable, and two ends of each pipeline are open; and the pipelines are disposed in the box body by a pipe frame, and a spacing is formed between the lower end opening of each pipeline and the bottom part of the box body. The method for the batch identification of wheat salt tolerance with application of the device comprises the steps that (1) soil is put in the pipelines in the device; (2) seeds of to-be-identified wheat are planted in the pipelines; (3) the pipelines are placed in the box body by the pipe frame, wherein a nutrient solution is stored in the box body, and the lower ends of the pipelines are soaked in the nutrient solution; and (4) the batch identification of the wheat salt tolerance can be realized through had justment and control of the salt content of the nutrient solution. The method is widely applicable to the salt tolerance identification of wheat germplasm resources, genetic materials and new varieties (species).
Description
Technical field
The present invention relates to a kind of method and device of plant identification salt resistance, be specifically related to method and the device of a kind of batch qualification salt tolerance of wheat.
Background technology
The soil salinization is a global resource and ecological problem.According to statistics, whole world saline soil area is 9.5438 hundred million hectares, and China's saline soil is widely distributed, and the gross area is 14.87 hundred million mu (being roughly equal to 100,000,000 hectares).Appropriate application salinization soil resource is a problem in the urgent need to address in China's agricultural development.The development and utilization of salinization soil has number of ways, and plantation salt tolerant crop kind is one of cost-effective approach.Wheat is second largest cereal crops in the world, simultaneously one of the Main Cultivation crop of Ye Shi China salinization soil.The soil salinization has a strong impact on wheat yield, and therefore, seed selection Salt-tolerant Wheat kind and research wheat salt tolerance characteristic are the important channels effectively utilizing salinization soil resource, improve grain yield.
The salt-tolerant trait of wheat is by the quantitative character of multipath, controlled by multiple genes, and the salt resistance of different developmental stages also there are differences.In the whole growth cycle of wheat, bud phase, seedling stage and bloom-be in the milk-maturing stage is especially responsive to salinity.In general, wheat malt phase salt resistance refer to seed imbibition under condition of salt stress, the integration capability of taking root of sprouting.Wheat Seedling salt resistance is then that under the soil accumulation of salt in the surface soil phase in spring, winter wheat turns green and resists the ability of salt stress after spring wheat insemination and emergence.Salt tolerance of wheat reduces output economical character reduces degree ability by salt stress.In general, the Salt-Tolerance Identification in wheat malt phase and seedling stage is wheat salt tolerance individuality and the basis of strain Seedling selection, is undertaken by the Morphologic Characters and physiological parameter measuring wheat seedling under water planting Salt Strees Condition condition.If successively Salt-Tolerance Identification can be carried out to wheat malt phase and seedling stage under a kind of condition, then must substantially reduce the test period, improve test efficiency.
By culture medium classification, what salt tolerance of wheat was tested is mainly divided into earth culture and water planting two type.Earth culture experiment often needs salt pond, thus needs space comparatively large, spends high.The key facility of solution culture is relatively simple, comprises the plastic box for holding nutrient solution, for supporting the vacuolar membrane plate with holes of wheat growth and the air pump for leading to oxygen to nutrient solution.Salt tolerant handling procedure treats that wheat seeds sprouting grows to a leaf after date and enters in vacuolar membrane plate hole with sponge implant, swim in the culture fluid of plastic box, air pump is used to inflate in nutrient solution, in nutrient solution, add NaCl subsequently carry out Salt Strees Condition, the observation growth of cereal crop seedlings shows, and carries out investigation and the mensuration of Morphologic Characters and physical signs.Although water planting Tolerant salt can make wheat seedling occur salt stress phenotype fast, partly can reflect the Salt Tolerance at Seedling Stage characteristic of wheat, single water planting condition cannot simulate normal Wheat Seedling growing environment, also there is many deficiencies.Such as, because seedling basal growth is in vacuolar membrane plate hole, narrow space can limit tillering of wheat and grow.Further, due to once after being transplanted into vacuolar membrane plate wheat seedling cannot change planting location, thus cannot eliminate the difference of the condition of culture such as illumination and temperature, ensure the uniformity of each individual plant experimental condition.In addition, the step of the method is relatively loaded down with trivial details, needs to transplant seedling, can for want of support and lodge after seedling grows up before water planting, and must again transplant could breeding under earth culture condition.In addition, the equipment such as air pump are also needed to ensure there is enough oxygen in nutrient solution by ciltivating process qualification Wheat Seedling salt resistance.
Summary of the invention
The object of this invention is to provide method and the device of a kind of batch qualification salt tolerance of wheat, the inventive method utilizes soil-ciltivating process qualification salt tolerance of wheat, can not only Selection of Salt-Tolerant wheat be individual rapidly, and genetic analysis can be carried out to the salt resistance of wheat population, there is good novelty, practicality and popularity.
The device of batch qualification salt tolerance of wheat provided by the present invention, comprises the box body of a upper end open;
Be provided with several pipelines in described box body, and the position of described pipeline is adjustable, the two ends of described pipeline are that opening is arranged; Described pipeline is located in described box body by pipe support, and is provided with spacing between the bottom of the lower ending opening of described pipeline and described box body.
In the device of above-mentioned batch qualification salt tolerance of wheat, described pipeline can be plastic tube; Described box body can be plastic casing.
In the device of above-mentioned batch qualification salt tolerance of wheat, described pipeline is conical pipe, and its osculum end is arranged downwards, and native surface is beneficial to ventilative.
In the device of above-mentioned batch qualification salt tolerance of wheat, described pipe support is porous plate, and concrete hole count can be selected as required, as selected 98 orifice plates (7 × 14).
Invention further provides the method utilizing said apparatus batch qualification salt tolerance of wheat, comprise the steps:
(1) to filling soil in the described pipeline in the device of described batch qualification salt tolerance of wheat;
(2) seed of wheat to be identified is planted in the soil in described pipeline;
(3) by described pipe support, described pipeline is positioned in described box body; Fill nutrient solution in described box body, the lower end of described pipeline is dipped in described nutrient solution;
(4) by the salt content in the described nutrient solution of regulation and control, namely realize identifying the batch of salt tolerance of wheat.
In above-mentioned method, the salt content of described soil can be 0.5%.
In above-mentioned method, the degree of depth that the lower end of described pipeline is dipped in described nutrient solution can be 4.0 ~ 5.0cm.
In above-mentioned method, the salt content of described nutrient solution is 6g/L ~ 9g/L, and described culture fluid can select conventional wheat cultivation culture fluid as required.
In above-mentioned method, in step (2), the method for being planted by the seed of wheat to be identified in soil in described pipeline is: before plantation wheat, be first by the seed germination of wheat, then to plant in the pipeline after soaking in soil.
In above-mentioned method, the degree of depth of the soil of filling in described pipeline can be 19 ~ 20cm.
In above-mentioned method, in described step (4), also comprise the steps: the following index of the wheat detecting bud phase and/or seedling stage: K in the upperground part biomass, root length, cambium
+ion concentration and Na
+ion concentration, judges the salt resistance of wheat by described index, top biomass and root higher still can be kept long, maintain high K in cambium under condition of salt stress
+ion concentration, low Na
+the individuality of ion concentration is that salt tolerant is individual.
The inventive method takes soil-water culture to carry out the qualification of salt tolerance of wheat.
The inventive method is high by the way simulation beach saline land subterranean water level of adding sea salt in nutrient solution, the natural characteristic that the saline and alkaline degree of water is high.The present invention comprehensively uses soil, water two kinds of different medium seed selection Salt-tolerant Wheats, both the homogeneity of salting liquid intermediate ion concentration had been ensured, salinity vertical distribution in soil can be ensured again, and the morphological character of wheatland top and underground part and physical signs under salt stress can be investigated easily.Wheat seed is directly sown into and is equipped with in native plastic tube by the inventive method, whole breeding time can be completed under the condition that need not transplant, thus not only Wheat Seedling salt resistance can be investigated, and can to early to the wheat malt phase, evening to wheat flower-grouting-maturing stage carries out Salt-Tolerance Identification, has very strong practicality.In addition, because plastic tube can put the optional position on porous pipe support, therefore, it is possible to periodically reversing of position easily, ensure the uniformity of the experimental condition such as illumination and temperature.In addition, when utilizing this method to carry out wheat salt tolerance qualification, root system can not anoxic in nutrient solution, does not therefore need the device of air pump.This method is widely used in the Salt-Tolerance Identification of Wheat Germplasm Resources, genetic stocks, new lines (kind).
Accompanying drawing explanation
Fig. 1 is the structural representation of the device of the present invention's batch qualification salt tolerance of wheat.
In figure, each mark is as follows: 1 plastic casing, 2 pipe supports, 3 plastic tubes.
Fig. 2 is the photo utilizing apparatus of the present invention to carry out soil-water culture experiment qualification Wheat Seedling salt resistance.
Fig. 3 is medium and small 60 and the phenotypic difference of Jimai 22 under contrast and Ficus caricaL of laying down of embodiment 2.
Fig. 4 is that in embodiment 2, under contrast and Ficus caricaL condition, little 60 and the overground part dry weight of Jimai 22 and root of laying down are long.
Fig. 5 is the different tissues K of little lay down 60 and Jimai 22 under contrast and Ficus caricaL condition in embodiment 2
+concentration, Na
+concentration, K
+/ Na
+concentration ratio.
Embodiment
The experimental technique used in following embodiment if no special instructions, is conventional method.
Material used in following embodiment, reagent etc., if no special instructions, all can obtain from commercial channels.
The device of embodiment 1, batch qualification salt tolerance of wheat
As shown in Figure 1, for the structural representation (illustrate only the setting of the 1st row and the 1st row plastic tube) of the device of batch qualification salt tolerance of wheat provided by the invention, it comprises the plastic casing 1 (long 65cm × wide 40cm × high 16cm) of a upper end open, pipe support 2 (the Stuewe & Sons company of the U.S. of 98 hole (7 × 14) is set up at this plastic casing 1, model: RL98), (its nozzle diameter is 3.8cm to the plastic tube 3 of a placement taper in each hole, length is 21cm, volume is 164ml, Stuewe & Sons company of the U.S., model SC10), the both ends open of plastic tube 3, its osculum end is arranged downwards, and be provided with spacing between the bottom of the lower ending opening of plastic tube 3 and plastic casing 1, this spacing can carry out regulating (spacing is about 1cm) according to different requirement of experiment.
In apparatus of the present invention, can according to different identification experiment requirement, the size of plastic casing 1, the number of plastic tube 3 and internal diameter are set and length is selected according to specific needs.
The salt resistance of embodiment 2, qualification wheat
The present embodiment select wheat breed little lay down 60 and Jimai 22 as examination material.
Wheat breed Jimai 22 is the Super-high-yielding of the up-to-date incubation of Crop Inst. of shandong Prov. Agriculture science Academy, many anti-, Medium gluten wheat new varieties, in September, 2006 and in January, 2007, authorization numbering was respectively Lu Nong careful 2006050 and state examines wheat 2006018 respectively by Shandong Province and the sheet authorization of national the Yellow River and Huai He River north.
The new variety of wheat of the drought resisting that wheat breed little Yan60Shi Inst. of Genetics and Development Biology, CAS cultivates, salt tolerant, high yield, high-quality, has applied for and has obtained New variety protection power, application number: 20100586.6.
Soil is filled out by in-built for the plastic tube 3 in embodiment 1 device, and nutrient solution is added in plastic casing 1, nutrient solution is according to document (Ren Yongzhe, Xu Yanhua, expensive auspicious the .2012 salt stress such as to defend under regulate and control Wheat Seedling proterties qtl analysis. Scientia Agricultura Sinica .45:2793-2800.) in formulated.This device is utilized to carry out the qualification of salt resistance.
One, for the salt stress process of examination material
Salt Tolerance at Seedling Stage phenotypic evaluation is carried out by soil-water culture.
Select the wheat seed of uniformity, 10% hydrogen peroxide sterilizing 30 minutes, deionized water rinsing for several times, evenly being placed in by seed is covered with in the culture dish of filter paper, sprout 2 days in 25 DEG C, then select to sprout consistent seed, ventral groove is seeded in downwards in the plastic tube 3 that soil is housed.In order to the soil in wet tube, before sowing, plastic tube 3 is put into the pipe support 2 in 98 holes (7 × 14), pipe support 2 is placed in the plastic casing 1 filling 10 liters of nutrient solutions.
Salt stress two process of salt adding control treatment and soil salt content 0.5%, does not after planting start to carry out Ficus caricaL.The each plastic casing 1 of control treatment adds the nutrient solution of 10L containing 0g/L salt, salt processing method: be increased to 6g/L after 3 days from 3g/L process, and reprocessing was increased to 9g/L after 6 days, keeps this concentration (salt content of simulation coastal saline-alkali soil water) later.Each process repeats for 7 times.The each plastic casing 1 of Ficus caricaL adds the nutrient solution of 10L saliferous.Condition of culture is: cultivation temperature is that between 20 DEG C of (daytime)/10 DEG C (night), the photoperiod is 12h/12h.Within every 3 days, change one time of nutrition liquid.In each test, each plastic tube 3 and each plastic casing 1 are put at random, and regular reversing of position (Fig. 2).Retain the plastic tube 3 do not sowed in test as blank, measure as soil salt.
Two, soil salt content measures
When salt stress wheat wheat seeding 21 days, random selecting 3 blanks, point get 3 soil sample layers (0-5cm, 5-10cm and 10-15cm), evenly fetch earth, then mix.Carry out air-dry, by V to gathered soil sample
soil: V
water=1:5 carries out lixiviate.Adopt the Main Anions (HCO of chemical titration leaching liquor
3 –, Cl
–, SO
4 2 –) and cation (Ca
2+, Mg
2+, K
++ Na
+) content, zwitterion content sum is soil salt content.
Soil salt content is that the method recorded in bibliography (huge the last million, Liu little capital .2012. arid RS in Alkali Region tillage method changes affects .16 (7): 6-10. to soil property and crop yield) measures.Wherein, K
+, Na
+, Ca
2+, Mg
2+measure with atomic absorption spectrophotometer; SO
4 2 –eDTA titration is adopted to measure, Cl
–silver nitrate titration method is adopted to measure; HCO
3 –double-tracer technique is adopted to measure.
In different soil soil main zwitterion content and total salometry result as shown in table 1.
Main zwitterion content and total salometry result in table 1 different soil soil
As can be seen from the data in table 1, in each component forming soil salt, anion is mainly with Cl
–be main.Then there is the change between different soil in cation, at 0-5cm soil layer mainly with Ca
2+be main, with K in 5-10cm soil layer and 10-15cm soil layer
++ Na
+be main.In addition, different soils layer salinity vertical change is obvious, and soil salt content is the highest in 0-5cm soil layer, reaches 0.5%; Take second place in 10-15cm soil layer, reach 0.3%; Lower slightly in 5-10cm soil layer, be 0.23%.Utilize apparatus of the present invention to carry out Salt-Tolerance Identification, soil salt content reaches 0.23%-0.5%, substantially simulates the gentle distribution trend of soil brackish water in moderate salination district, strand.
Three, for the Salt-Tolerance Identification of examination material
Salt stress wheat wheat seeding measured phenotypic character after 23 days, measured characteristic index and comprised: the upperground part biomass, root are long, overground part Na
+and K
+ion concentration.
Before wheat plant is gathered in the crops, add up every strain root long, claim fresh weight during wheat plant results, complete 15 minutes in 105 DEG C after results, dry to constant weight for 70 DEG C, claim dry weight.Dry sample comminution, carry out microwave with nitric acid-hydrogen peroxide and disappear and boil, measure overground part Na
+and K
+content (measuring with inductively-coupled plasma spectrometer (ICP-OEC)) and calculating K
+/ Na
+than (Wang little Ping, Xiang Su stay .2006. micro-wave digestion-ICP-OES, AAS and AFS measures garlic different parts 20 kinds of constituent content .26 (10): 1907-1911.).
Little lay down 60 and Jimai 22 contrast and Ficus caricaL under phenotypic difference as shown in Figure 3, as seen from Figure 3, little lay down 60 and Jimai 22 be all suppressed in the growth of Ficus caricaL its root system and overground part after 23 days.Compared with the control, the upperground part biomass of latter two kind of Ficus caricaL and root are long all significantly to be reduced, little lay down 60 and Jimai 22 overground part dry matter be 0.37g and 0.22g respectively, only have 49.5% and 45.1% of contrast respectively; Little lay down 60 and Jimai 22 long be 28.9cm and 10.2cm respectively, only have 71.3% and 23.6% (as shown in Figure 4) of contrast respectively.After Ficus caricaL, the long and relative value of the biomass of Jimai 22 and root is all starkly lower than littlely lays down 60.
Na
+ion toxicity is that plant salt coerces one of downtrod influence factor of rear growth.Improve K
+absorb, suppress Na
+high K in interior stream, maintenance cell
+/ Na
+than being one of important research content improving plant salt tolerance.For analyzing the physiological mechanism of little lay down 60 and Jimai 22 salt resistance difference, the present invention determines for the K of examination material under contrast and Ficus caricaL condition in different tissues
+ion and Na
+ion concentration, and calculating K
+/ Na
+concentration ratio.
The present invention compares for trying K between the different tissues of material under contrast and Ficus caricaL condition
+the difference of ion concentration.As shown in Fig. 5 (a), under collating condition, two Ks of kind in different tissues
+ion concentration does not have significant difference.Ficus caricaL is to K in different tissues
+there is Differences in the impact of ion concentration.After Ficus caricaL, the K in two kind root systems
+ion concentration reduces, the K in stem
+ion concentration change is not obvious; Lao Ye K
+ion concentration does not have significant change little laying down in 60, obviously reduces, young leaves K in Jimai 22
+ion concentration obviously increases little laying down in 60, obviously reduces in Jimai 22, shows little 60 K in young leaves that lay down
+ion concentration will apparently higher than Jimai 22.K in little 60 blades of laying down after Ficus caricaL
+ion concentration higher than Jimai 22, K in body
+ion concentration variable effect is not obvious, and Ficus caricaL significantly reduces K in Jimai 22 body
+ion concentration.Illustrate that salt stress seriously inhibits Jimai 22 couples of K
+absorption and transhipment to overground part blade, and little lay down 60 substantially unaffected.
As shown in Fig. 5 (b), Ficus caricaL significantly increases the Na in different tissues
+ion concentration, but the increasing degree of Wheat Cultivars there are differences.Under collating condition, two Nas of kind in different tissues
+ion concentration does not have notable difference, but under Ficus caricaL, little 60 Na in young leaves that lay down
+ion concentration is starkly lower than Jimai 22, and the concentration of two kinds in root system, Lao Ye and stem does not have significant difference.Above-mentioned data illustrate, lay down compared with in the of 60 with little, the Na that under Ficus caricaL, Jimai 22 absorbs
+be transported to more in young leaves.
As shown in Fig. 5 (c), Ficus caricaL significantly reduces the K in 4 different tissues
+/ Na
+ratio, but to the K in Jimai 22 young leaves and stem
+/ Na
+the impact of ratio be greater than on little lay down 60 impact, show the little K laid down in 60 young leaves and stem
+/ Na
+ratio is higher than Jimai 22 (Fig. 5 (d)).Medium and small 60 K that can keep higher that lay down of cambium after the above results explanation Ficus caricaL
+/ Na
+ratio, is conducive to it and improves salt resistance.
The present invention, by under contrast and condition of salt stress, measures for little 60 and the partial phenotypic of Jimai 22 and the physical signs of laying down of examination material, infers practicality and the reliability of this invention.By salt stress process, two kind root systems and overground part growth all comparatively contrast significantly reduce, K in root system
+ion concentration reduces, Na in different tissues
+ion concentration and K
+/ Na
+than significantly increasing, illustrate that the salt stress process using the present invention to carry out Wheat Cultivars is general effective.In addition, after salt stress process, compared with Jimai 22, little lay down 60 biomass and root is long all obviously increases, young leaves intermediate ion concentration increases, Na
+ion concentration reduces, and keeps higher K
+/ Na
+ratio, illustrates little 60 comparatively Jimai 22 more salt tolerants of laying down.Therefore illustrate and use the present invention can the salt resistance of precise Identification Wheat Cultivars.
Utilize the present invention can use same device successively qualification wheat malt phase and Salt Tolerance at Seedling Stage characteristic, Selection of Salt-Tolerant wheat is individual rapidly, under condition of salt stress, top biomass and root higher namely still can be kept long, maintain high K in cambium
+ion concentration, low Na
+the individuality of ion concentration, be a kind of technological break-through, therefore the inventive method has good novelty, practicality and popularity.
Claims (10)
1. a device for batch qualification salt tolerance of wheat, is characterized in that: described device comprises the box body of a upper end open;
Be provided with several pipelines in described box body, and the position of described pipeline is adjustable, the two ends of described pipeline are that opening is arranged; Described pipeline is located in described box body by pipe support, and is provided with spacing between the bottom of the lower ending opening of described pipeline and described box body.
2. device according to claim 1, is characterized in that: described pipeline is plastic tube; Described box body is plastic casing.
3. device according to claim 1 and 2, is characterized in that: described pipeline is conical pipe, and its osculum end is arranged downwards.
4. the device according to any one of claim 1-3, is characterized in that: described pipe support is porous plate.
5. a method for batch qualification salt tolerance of wheat, comprises the steps:
(1) to filling soil in the described pipeline in device according to any one of claim 1-4;
(2) seed of wheat to be identified is planted in the soil in described pipeline;
(3) by described pipe support, described pipeline is positioned in described box body; Fill nutrient solution in described box body, the lower end of described pipeline is dipped in described nutrient solution;
(4) by the salt content in the described nutrient solution of regulation and control, namely realize identifying the batch of salt tolerance of wheat.
6. method according to claim 5, is characterized in that: the salt content of described soil is 0.5%; The salt content of described nutrient solution is 6g/L ~ 9g/L.
7. the method according to claim 5 or 6, is characterized in that: the lower end degree of depth be dipped in described nutrient solution of described pipeline is 4.0 ~ 5.5cm.
8. the method according to any one of claim 5-7, it is characterized in that: in step (2), the method that the described seed by wheat to be identified is planted in the soil in described pipeline is: first by the seed germination of wheat, then to plant in the pipeline after wetting in soil;
In described step (4), also comprise the steps: the following index of the wheat detecting bud phase and/or seedling stage: K in the upperground part biomass, root length, cambium
+ion concentration and Na
+ion concentration, judges the salt resistance of wheat by described index.
9. the method according to any one of claim 5-8, is characterized in that: the degree of depth of the soil of filling in described pipeline is 19 ~ 20cm.
10. the application of device according to any one of claim 1-4 in batch qualification salt tolerance of wheat.
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| CN110313268A (en) * | 2019-07-16 | 2019-10-11 | 天津绿茵景观生态建设股份有限公司 | A method of utilizing culture apparatus Rapid identification seed salt tolerance |
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