CN106665311A - Method for enhancing cadmium tolerance of hibiscus cannabinus seedlings - Google Patents
Method for enhancing cadmium tolerance of hibiscus cannabinus seedlings Download PDFInfo
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- CN106665311A CN106665311A CN201611258458.2A CN201611258458A CN106665311A CN 106665311 A CN106665311 A CN 106665311A CN 201611258458 A CN201611258458 A CN 201611258458A CN 106665311 A CN106665311 A CN 106665311A
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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
- A01G31/00—Soilless cultivation, e.g. hydroponics
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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
- A01G22/00—Cultivation of specific crops or plants not otherwise provided for
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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
- A01G24/00—Growth substrates; Culture media; Apparatus or methods therefor
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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
- A01G7/06—Treatment of growing trees or plants, e.g. for preventing decay of wood, for tingeing flowers or wood, for prolonging the life of plants
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N37/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids
- A01N37/44—Biocides, pest repellants or attractants, or plant growth regulators containing organic compounds containing a carbon atom having three bonds to hetero atoms with at the most two bonds to halogen, e.g. carboxylic acids containing at least one carboxylic group or a thio analogue, or a derivative thereof, and a nitrogen atom attached to the same carbon skeleton by a single or double bond, this nitrogen atom not being a member of a derivative or of a thio analogue of a carboxylic group, e.g. amino-carboxylic acids
- A01N37/46—N-acyl derivatives
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
- A01N43/00—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds
- A01N43/02—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms
- A01N43/04—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom
- A01N43/14—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings
- A01N43/16—Biocides, pest repellants or attractants, or plant growth regulators containing heterocyclic compounds having rings with one or more oxygen or sulfur atoms as the only ring hetero atoms with one hetero atom six-membered rings with oxygen as the ring hetero atom
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- General Health & Medical Sciences (AREA)
- Pest Control & Pesticides (AREA)
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- Agronomy & Crop Science (AREA)
- Biodiversity & Conservation Biology (AREA)
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- Agricultural Chemicals And Associated Chemicals (AREA)
Abstract
The invention discloses a method for enhancing cadmium stress tolerance of hibiscus cannabinus seedlings, belongs to the field of plant cultivation, and aims to provide a method for remarkably enhancing soil cadmium stress tolerance of the of hibiscus cannabinus. Hibiscus cannabinus has the characteristics of short growing period, high biomass (as three to four times of that of trees), high heavy metal tolerance, wide adaptability and the like, and as one of fiber crops, the hibiscus cannabinus is less prone to carrying heavy metal into a food chain, so that the hibiscus cannabinus is expected to remedy large areas of soil polluted by the heavy metal, and is capable of not only remedying soil pollution, but also capable of obtaining high economic benefits. Aiming to remarkably enhancing soil cadmium stress tolerance of hibiscus cannabinus, the invention further discloses a preparation method of an applied composition.
Description
Technical field
The invention belongs to field of plant cultivation, and in particular to the method for improving the resistance to cadmium performance of ambary Seedling Stage.
Background technology
Ambary has that growth cycle is short, Biomass big (for the three-to-four-fold of trees), preventing from heavy metal ability are strong, wide adaptability
The features such as, and because being fibre crops, be difficult to bring heavy metal into food chain.Therefore, ambary be expected to be used for administer large area receive
The soil of heavy metal pollution, can not only administer soil pollution, can also obtain higher economic benefit.Qi Jianmin etc. have studied
The Absorption Characteristics of ambary heavy metal, have filtered out high resistance to, high cadmium Kenaf Cultivars, have studied ambary heavy metal cadmium
Uptake and accumulation feature and its interracial difference；The antioxidant system for comparing high resistance to kind and low resistance to kind is responded to Cd stress
The impact to ambary root-tip cells structure of difference and Cd stress (Absorption Characteristics of ambary heavy metal and external source GSH are alleviated
The study mechanism of cadmium poison, University Of Agriculture and Forestry In Fujian Ph.D. Dissertation, 2013).
The research such as Deng Yong finds that ambary young root malonaldehyde, As A contents and stress time and cadmium concentration are presented positive correlation.As
The change of A and GSH contents and activities of antioxidant enzymes is probably the important physiological foundation (Cd stress that ambary resists cadmium poisoning and resistance
The physiological responses of lower ambary young root, China's fiber crops industry science, 2016).Li Lanping is by Study On Lead, cadmium to kenaf seeds germination and growth
Impact, using the concentration of Cd≤20mg/kg as distinguishing the strong and weak ideal concentration of the resistance to cadmium of Kenaf Cultivars.
Although the characteristic of the resistance to cadmium of ambary has been widely recognized, but extensively carry out planting also as rehabilitation plant
It is not promoted, inventor is had found by furtheing investigate, the potentiality of ambary root absorption soil heavy metal cadmium also have room for promotion,
The key position of phytoremediation is root system, and Cd is so that symplast is into the tip of a root and is deposited on epidermis cell, then is transported to by root system
Other positions.Ambary is flourishing root system better than the main cause of other industrial crops, in order to strengthen ambary heavy metal Cd
Absorbability, inventor has done various trials, for example, extend ambary main root length, increases fibrous root quantity etc., but produces little effect,
How to excite ambary to absorb soil heavy metal Cd, be the key link that ambary develops into heavy metal soil remediation crop.
Inventor through long-term research and cultivation practices, kenaf seeds are sprouted and growth of seedling during Cd stress enter
Gone further investigation, found ambary in whole trophophase, the period most sensitive to Cd stress has been Seedling Stage, especially sowing or
30-60 days after transplanting, once ambary seedling smoothly spends this sensitive period, the shadow of the Cd stress of same concentrations to its growth promoter
Sound will not result in its death, and the purpose of the present invention is exactly further to improve between Their Seed Germinating Period and during seedling development
Cadmium toleration.
The content of the invention
The purpose of the present invention is the performance for improving ambary Seedling Stage tolerance soil every stress, to reach this purpose, the present invention
There is provided a kind of method of raising ambary Seedling Stage heavy metal tolerance cadmium.
The method of the invention is related to a kind of compositionss, therefore the present invention also by the preparation method of open said composition.
The compositionss by five class compound groups into, five class compounds be respectively S-nitrosoglutathione glutathion (GSNO),
Cellulose derivative, catechin, surfactant and chelating agen.
GSNO is obtained by nitrite (NO2-) and reduced glutathion (GSH) reaction, and GSNO diluents are in compositionss
Middle content is 1.0-1.5%.
Cellulose derivative is in compositionss：Methylcellulose (Mc), hydroxymethyl cellulose (HMc), hydroxypropyl methyl are fine
In dimension plain (HPMC), hydroxyethylmethyl-cellulose (HEMC), methylethylcellulose (MEC) and methylpropylcelluloses (MPC)
At least one cellulose derivative.
In the composition weight/mass percentage composition is 65.5-86.5% to cellulose derivative.
Catechin in compositionss refers to the non-phenotype youngster such as catechin, nutgall catechin, catechin and gallate
In theine class, and epicatechin, epigallo catechin, L-Epicatechin gallate isophenous catechin at least one
Kind.
In the composition weight/mass percentage composition is 3.5-8.5% to catechin
Surfactant in compositionss, refers to sodium alkyl benzene sulfonate, sodium alkyl sulfonate, Negel, 2- naphthalene sulphurs
One or more in the condensation substance salt of the horizontal acid of sour sodium, naphthalene.
In the composition weight/mass percentage composition is 5.5-10.5% to surfactant.
Chelating agen in compositionss refers to the degradable organic acid such as aminotriacetic acid (NTA) or citric acid (CA).
In the composition weight/mass percentage composition is 1.5-2.5% to chelating agen.
In compositionss in addition to GSNO, remaining compound is commercially available.
The preparation method of compositionss is as follows：
(1) cellulose derivative of certain mass percent, catechin, surfactant and chelating agen are dissolved in ethanol；
(2) the GSNO diluents of certain mass percent are added in solution in step (1)；
The using method of the compositionss that the present invention is provided is as follows：
(1) certain density GSNO mother solutions are added in nutritional solution during water planting seedling, the concentration for making GSNO in culture fluid is
125-300 μm of ol/L, transplants to land for growing field crops, applies the compositionss containing GSNO diluents.
(2) manage after transplanting：Ambary seedling is conventionally transplanted and manages, and carries out foliage-spray within 30 days after transplanting,
The preferred 150-1500ppm of composition concentration, the preferred 350-500ppm of further concentration, amount of application is 10-50Kg/ mu soil；Move
Carry out within 60 days after cultivation foliage-spray again, the preferred 150-1500ppm of composition concentration, the preferred 200-800ppm of further concentration.
Amount of application is 20-60Kg/ mu soil.
The invention has the beneficial effects as follows, by using this method, under Cd stress, the suppression of ambary root system and aerial partss
Substantially it is eased, leaf development recovers normal, and yellowing leaf degree is obviously reduced, and photosynthesis gradually recover, and lateral root starts
Restoration ecosystem.
Specific embodiment
It is prepared by the GSNO mother solutions of embodiment 1
The preparation of GSNO mother solutions:Isopyknic 10mmol/LNaNO2(in 100mmol/LHCl) with it is isopyknic
10mmol/LGSH is (in the Na of 100mmol/LpH7.2HPO4- NaH2PO4In phosphate buffer) reaction, you can form 5mmol/
LGSNO (mixed liquor includes 5mmol/LGSNO).
It is prepared by the GSNO diluents of embodiment 2
GSNO mother solutions are stored on ice, stand-by with pure water dilution ten times (i.e. 500 μm ol/L) when using.Under room temperature,
(50 μm of ol/LDTT are included in the phosphate buffer of 100mmol/LpH7.4), GSNO can just discharge NO, wherein 500 μm of ol/
The GSNO 30min of L about discharge 3.25 μm of ol/L of NO.
The preparation of the compositionss of embodiment 3
By 78.5 grams of hydroxymethyl cellulose (HMc), 7.5 grams of nutgall catechins, 10.5 grams of 2- sodium naphthalene sulfonates and 2.5 grams of ammonia
Base triacetic acid is dissolved in 200mL ethanol, is added thereto to 1 gram of GSNO diluents (concentration is 500 μm of ol/L), and resulting solution is this
The compositionss that invention is provided.
The preparation of the compositionss of embodiment 4
By 86.5 grams of hydroxypropyl methyl celluloses (HPMC), 4.5 grams of catechin and gallates, 6 grams of Negels
Be dissolved in after 200mL ethanol with 1.5 grams of aminotriacetic acids, be added thereto to 1.5 grams GSNO diluents (concentration be 500 μm of ol/
L), the compositionss that resulting solution is provided for the present invention.
The using method of the compositionss of embodiment 5
Germination test:The complete seed of full and embryo is selected, 3%H is used2O2Sterilization 15min, distilled water rinse repeatedly after
Soaked overnight in distilled water so as to fully imbibition.Then seed is rested on into bottom to be covered with the culture dish of moistening filter paper, culture dish
Being put in 25 DEG C of constant temperature light cultures makes germination in 48 hours.
Seedling is cultivated：After bud grows to 2~3cm (about 3~4d), (intensity of illumination is 800 μ to be placed in culture between illumination cultivation
mol·m-1·S-1, photoperiod 14h/10h, 28 DEG C/20 DEG C of diurnal temperature, relative air humidity 70%).Training is adapted in tap water
After foster 3d, will cultivate in seedling replanting to 1/4Hoagland nutritional solutions, and the 5mmol/L GSNO mother solutions of above-mentioned synthesis are added
To in Hoagland nutritional solutions so as in GSNO concentration be 125-300 μm of ol/L.
Hoagland nutrient solution prescriptions are Ca (NO3) 25.79mM, KNO38.02mM,NH4H2PO41.35mM,
MgSO44.17mM,MnSO48.90 μM, H3BO348.3μM,ZnSO40.94μM,CuSO40.20μM,(NH4)2MoO40.015μM,
Fe-EDTA72.6 μM, with NaOH pH6.5 or so is adjusted.
Manage after transplanting：Ambary seedling is conventionally transplanted and manages, and carries out foliage-spray within 30 days after transplanting, combination
The preferred 150-1500ppm of thing concentration, the preferred 350-500ppm of further concentration, amount of application is 10-50Kg/ mu soil.
Foliage-spray is carried out again within 60 days after transplanting, the preferred 150-1500ppm of composition concentration, further concentration is preferred
200-800ppm.Amount of application is 20-60Kg/ mu soil.
Embodiment 6 uses the index determining of the ambary seedling of this compositions
1. thick the index plant height of seedling aerial partss, stem, leaf area and Herb biological quality are determined, for indication ambary
Growth performance.
Under different components concentration (0-850ppm) process, to Cd stress, (cadmium concentration that test sets is native as 200mg/Kg
Earth) under potted plant seedling physiology form impact, test sets matched group, and matched group is the life of ambary seedling without under Cd stress
Long situation, the trophophase of all test plants is 30 days, and the test quantity of every group of ambary is 20 plants；
Using this compositions, Cadmium in Soil has obtained notable alleviation to the stress of ambary growth of seedling, and the compositionss of optimization are dense
Spend for 350-500ppm.
2. Contents of Photosynthetic Pigments adopts acetone extraction method, is detected with UV-2450 types ultraviolet-uisible spectrophotometer, exists respectively
Mensuration absorbance value at wavelength 663,645 and 470nm, by formula the dense of chlorophyll a, chlorophyll b and carotenoid is calculated respectively
Degree, chlorophyll a is added with the concentration of chlorophyll b and obtains final product chlorophyll total concentration.
Under test is determined as variable concentrations compositions-treated, to ambary seedling Contents of Photosynthetic Pigments under soil Cd stress
Affect.Compositions-treated more than 150PPM makes the inhibitory action of each Contents of Photosynthetic Pigments of Cd stress Corchorus olitorius L. seedling significantly be alleviated
(P ＜ 0.05), and with the rising of compositions-treated concentration, mitigation gradually increases.Concentration makes for PPM350-500 process
Chlorophyll total amount, chlorophyll a, chlorophyll b and carotenoid content are respectively than for increased for being processed using said composition
290.98-307.74%, 210.86-254.55%, 438.65-621.01% and 95.65-109.31%, carotenoid contains
Amount has been even up to the level of control, i.e. inhibitory action is released from completely.
3. the determination of activity of antioxidase adopts the spectrophotography of Elavarthi and Mart in.
Trophophase be the potted plant ambary of 30 days under the Cd stress of 200mg/Kg soil, no matter in root system or leaf tissue
On, all significantly inhibit the activity of several antioxidases.Compared with the normal seedling without Cd stress, Cd stress process makes ambary
SOD, CAT, APX and POD activity of root system have dropped respectively 69.30%, 81.97%, 90.91% and 71.93%, blade
SOD, CAT, APX and POD activity have dropped respectively 34.86%, 78.68%, 53.16% and 57.90%.
The response of ambary seedling activities of antioxidant enzymes and the concentration of compositionss, pheron self-characteristic and tissue specificity are all
It is relevant.
The compositionss of suitable concentration are sprayed and process several antioxidases in the ambary seedlings root and blade made under Cd stress
Activity is dramatically increased (P ＜ 0.05).SOD, APX activity increases with composition concentration and increases in ambary leaf tissue,
The process of PPM445 concentration reaches peak value, and SOD, APX activity of root system tissue reaches peak value, concentration for the treatment of in the process of PPM490 concentration
This level is kept at after increasing again.CAT, POD activity is with composition concentration in ambary leaf tissue and root system tissue
Increase and increase, the process of PPM500 concentration reaches peak value, illustrate with regard to compositionss induction for the anti-oxidative defense of Cd stress,
Sensitivity of the different antioxidases in root system with blade is different, also different to the sensitivity of composition concentration.Comprehensively use
Cost consideration, the concentration after optimization is 445-500PPM.
Impact of the using method of the compositionss of embodiment 7 to effect
Add the S-nitrosoglutathione glutathion (GSNO) of 125~250 μm of ol/L during water planting ambary seedling in nutritional solution,
And transplanting to land for growing field crops, apply certain density compositionss, can further improve the using effect of this compositions, concrete table
Now the suppression situation of ambary root system and aerial partss, compared with Seedling Stage sprays this compositions, is further eased with only,
Leaf development recovers rapidly normal, and yellowing leaf degree can be recovered, and photosynthesis are gradually normal, and lateral root starts to recover life
It is long.
Claims (6)
1. a kind of method for improving the resistance to cadmium performance of ambary Seedling Stage, including the preparation method containing GSNO compositionss and its user
Method, is characterised by, the preparation method of GSNO is as follows：
A:The preparation of GSNO mother solutions：Isopyknic 10mmol/L NaNO2With isopyknic 10mmol/L GSH, in 100mmol/L
The Na of pH7.42HPO4- NaH2PO4React in phosphate buffer, form the GSNO mother solutions that concentration is 5mmol/L, the storage of GSNO mother solutions
Exist on ice；
B：The preparation of GSNO diluents：Mother solution is diluted into ten times with pure water, the GSNO dilutions that concentration is 500 μm of ol/L are obtained
Liquid；
Preparation method containing GSNO compositionss is as follows：
C:By the cellulose derivative of mass percent 65.5-86.5%, the catechin of 3.5-8.5%, 5.5-10.5%
Surfactant and 1.5-2.5% chelating agen are dissolved in ethanol；
D:Mass percent is added in solution in step C for the GSNO diluents of 1.0-1.5%.
2. the method for improving the resistance to cadmium performance of ambary Seedling Stage according to claim 1, is characterised by, using described containing GSNO
Compositionss Seedling Stage ambary is dealt with, method is as follows：
A. germinate：The complete seed of full and embryo is selected, 3%H is used2O2Sterilization 15min, distilled water rinse repeatedly after in distilled water
Middle soaked overnight so as to fully imbibition, then rests on seed bottom and is covered with the culture dish of moistening filter paper, and culture dish is put in 25
DEG C constant temperature light culture makes germination in 48 hours；
B. seedling culture：After bud grows to 2~3cm, it is placed between illumination cultivation and cultivates, intensity of illumination is 800 μm of olm-1·S-1,
Photoperiod 14h/10h, 28 DEG C/20 DEG C of diurnal temperature, relative air humidity 70%, after culture 3d is adapted in tap water, by seedling
Transplant into 1/4Hoagland nutritional solutions, and 5mmol/L GSNO mother solutions are added in Hoagland nutritional solutions, be allowed to be formed
125-300 μm of ol/L GSNO concentration, cultivates to 5 true leaves and is fully deployed；
C. seedling management：Carry out within 30 days after transplanting foliage-spray compositionss, the preferred 150-1500ppm of composition concentration, further
The preferred 350-500ppm of concentration, amount of application is 10-50Kg/ mu soil；
Carry out within 60 days after transplanting foliage-spray again, the preferred 150-1500ppm of composition concentration, the preferred 200- of further concentration
800ppm, amount of application is 20-60Kg/ mu soil.
3. the method for improving the resistance to cadmium performance of ambary Seedling Stage according to claim 2, is characterised by, cellulose in compositionss
Derivant is：Methylcellulose, hydroxymethyl cellulose, hydroxypropyl methyl cellulose, hydroxyethylmethyl-cellulose, Methylethyl are fine
At least one cellulose derivative in dimension element and methylpropylcelluloses.
4. the method for improving the resistance to cadmium performance of ambary Seedling Stage according to claim 3, is characterised by, the catechu in compositionss
Plain class refers to the non-phenotype catechin such as catechin, nutgall catechin, catechin and gallate, and epicatechin, table
At least one in nutgall catechin, L-Epicatechin gallate isophenous catechin.
5. the method for improving the resistance to cadmium performance of ambary Seedling Stage according to claim 4, is characterised by, the surface in compositionss
Activating agent, in being sodium alkyl benzene sulfonate, sodium alkyl sulfonate, Negel, 2- sodium naphthalene sulfonates, the condensation substance salt of the horizontal acid of naphthalene
One or more.
6. the method for improving the resistance to cadmium performance of ambary Seedling Stage according to claim 5, is characterised by, the chelating in compositionss
Agent refers to the degradable organic acid such as aminotriacetic acid (NTA) or citric acid (CA).
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Application publication date: 20170517 |