CN105191938A - Cold-resisting agent for tobaccos as well as preparation and usage methods of cold-resisting agent - Google Patents
Cold-resisting agent for tobaccos as well as preparation and usage methods of cold-resisting agent Download PDFInfo
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Abstract
The invention discloses a cold-resisting agent for improving cold resistance of seedlings of tobaccos as well as preparation and usage methods of the cold-resisting agent. The cold-resisting agent comprises main components as follows: 1.15-2.30 g/L of glycine betaine, 5.0-10.0 mg/L of salicylic acid, 3.0-5.0 mg/L of abscisic acid and 600-1,000 mg/L of 5-aminolevulinic acid. The agent is sprayed to leaf surfaces in the seedling recovering period and the root extending period before outplanting of the seedlings of the tobaccos or after land transplantation of the seedlings of the tobaccos, the agent is sprayed after the original agent is diluted by 3-5 times, and 25-30 L of the agent is sprayed per mu. The agent is a cold-resisting leaf surface spray agent prepared from physiological activity regulating substances, can effectively improve the resistance of shoots (the seedlings) of the tobaccos to low-temperature adverse situation, can better cope with low-temperature damage of birth blockage (and even early blossoming) of the seedlings of the tobaccos due to frequent late spring coldness in main southern tobacco producing areas, has no adverse effect on the quality of flue-cured tobaccos and has bright application prospect in tobacco production.
Description
Technical field
The invention belongs to tobacco and resist Chilling stress, suppression " early blossoming " generation technique field, being specifically related to a kind of for improving the cold resistant anti-cryogen of tobacco seedling and preparation and application thereof.
Background technology
Tobacco be originate in semi-tropical happiness temperature crop (its growth optimum temperature be 25 ~ 28 DEG C, 10 ~ 13 DEG C stop growing, 1 ~ 2 DEG C of low temperature can make death of seedling), environmental temperature (especially low temperature) has significant impact to the fertility of tobacco, yield and quality.Cigarette district of China is widely distributed, various places temperature condition differs greatly, and as Main Tobacco-growing Regions In South " cold in the late spring " takes place frequently, causes the fertility of " also seedling stage, stretch the root phase " tobacco plant bad and bring out the generation of " early blossoming ", have a strong impact on yield of tobacco and quality, also govern the development that sound tobacco is produced always.At present, the Comprehensive Traits of the main breed such as K326, cloud and mist 87 that production is promoted is more excellent, but it is poor to the patience of low temperature environment.
Endanger this practical problem for " cold in the late spring " that take place frequently in puzzlement tobacco leaf production, be bred as germ plasm resource needed for Heat pretreatment new product of tobacco lack by genetic improvement, breeding cycle is long and need a large amount of human and material resources and financial resources, belongs to strategic act; And improved the Heat pretreatment of tobacco plant as sprayed tobacco Heat pretreatment conditioning agent by agronomic measures, then can yet be regarded as and tackle the effective measures and method that early spring in tobacco leaf production, " cold in the late spring " endangered.
Therefore, Study and Development suits the tobacco Heat pretreatment conditioning agent improving tobacco Heat pretreatment technical essential becomes problem demanding prompt solution in tobacco leaf production, significant to the safety in production of guarantee and stable tobacco leaf.
Summary of the invention
The object of the invention is to the blank making up tobacco chilling damage counter-measure technical field, a kind of cold-resistant composite foliar spray that can improve tobacco seedling Heat pretreatment is provided, and have that method of preparation and use is simple, successful, the advantage such as with low cost.
First object of the present invention is to provide the formula (constituent) of above-mentioned anti-cryogen.
A kind of Resistance In Tobacco cryogen is the solution comprising following density component: betain 1.15 ~ 2.30g/L, salicylic acid 5.0 ~ 10.0mg/L, abscisic acid 3.0mg ~ 5.0mg/L, 5-ALA 600 ~ 1000mg/L.
Comprise beet paper mill wastewater in described anti-cryogen and be preferably 1.15mg/L; Salicylic concentration is preferably 5.0mg/L; The concentration of abscisic acid is preferably 3.0mg/L; The concentration of 5-ALA is preferably 0.6g/L.
Described anti-cryogen, also containing glycerine, Vc, calcium salt and trace element.
Calcium salt in described anti-cryogen is the one in nitrate or calcium chloride, and the concentration of calcium ion is 600 ~ 1000mg/L; The concentration of the Vc in described anti-cryogen is 500 ~ 1000mg/L, the concentration of glycerine is 15 ~ 20ml/L; Trace elements iron 5 ~ 10mg/L, boron 5 ~ 10/Lmg, manganese 0.5 ~ 1.0mg/L, zinc 10 ~ 25mg/L, molybdenum 0.05 ~ 0.10mg/L is also comprised) in often liter of anti-cryogen.
One or more in 0.5ml plant leaf blade surface active agent tween 20, Tween 80, triton x-100 are comprised in often liter of anti-cryogen.
Another object of the present invention is to provide the preparation method of above-mentioned anti-cryogen.
The preparation method of anti-cryogen of the present invention, comprises the following steps successively:
(1) salicylic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution I;
(2) component betain, 5-ALA, Vc, glycerine, calcium salt and trace element are added to the water, are constantly stirred to it and dissolve acquisition solution II completely;
(3) abscisic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution III;
(4) solution I, III being joined respectively in solution II and to add surfactant, constantly stirring and make it fully mix, obtained anti-cryogen stoste of the present invention, dilutes during use again.
A further object of the invention is to provide the using method of above-mentioned anti-cryogen.
Described anti-cryogen is at cigarette seedling foliage-spray.Specifically before cigarette seedling goes out garden 5 ~ 7 or cigarette seedling field-transplanting after " also seedling stage ", " stretching the root phase " breeding time rain or shine sky foliage-spray at dusk.Spray after being diluted by anti-cryogen 3 ~ 5 times during use, every mu of Application is 25 ~ 30L.Require to be sprayed onto on tobacco plant blade tow sides uniformly, be as the criterion with the complete moistening whole blade of solution.The planting density of tobacco leaf production is generally 1100 ~ 1300 or 1300 ~ 1500 plants/acre (there is unified regulation in various places tobacco office).
Resistance In Tobacco cryogen of the present invention is for after cigarette transplantation of seedlings, and microthermal climate in early spring (" cold in the late spring ") is to the designed research and development of harmful effect being in " also seedling stage ", the fertility of " stretching the root phase " tobacco seedling.
On the one hand, when the reason that " cold in the late spring " weather causes seedling growth and development bad mainly includes its experience low temperature stress: membrane structure " impaired ", the permeability of (1) tobacco seedling increase, and in cell, soluble substance and electrolyte can passively to seepages outside film; (2) resistivity of tobacco seedling self to low temperature is weak, will cause its metabolism " disorder " when meeting with low temperature stress, and the compatible solute content such as soluble sugar, proline and betain reduce, and Heat pretreatment weakens; (3) tobacco seedling SCAVENGING SYSTEM OF ACTIVATED OXYGEN ability declines, the large biological molecule such as active oxygen direct oxidation damaging cells internal protein, enzyme, nucleic acid of enrichment; (4) enzyme of tobacco seedling leaf photosynthesis system reduction alive, the oxidized decomposition of chlorophyll, cause cigarette seedling plant photosynthetic capacity to decline.On the other hand, the anti-cryogen that the present invention adopts comprises the physiologically active of plant materials such as betain, salicylic acid, abscisic acid, 5-ALA, Vc, glycerine, the cytoplasma membrane of experience Chilling stress being coerced to tobacco plant has good protective effect, the compatible solute content that cigarette seedling reply Chilling stress is coerced can be increased, alleviate the decline that antioxidase is lived, reduce the accumulation meeting with active oxygen when Chilling stress is coerced, maintain the physiologic effect such as tobacco plant photosynthetic activity; In addition, as the stress resistance of the abscisic acid controllable enhancing plant of " resistance hormone ".
Therefore, the present invention considers to start with from above-mentioned many-side, the compound formulation of the physiologically active Auto-regulators such as modulation betain, salicylic acid, abscisic acid, 5-ALA, Vc, glycerine, through the process of cigarette seedling foliage-spray to improve the Chilling stress resistivity of tobacco plant, thus promote that the fertility status that experience " cold in the late spring " Chilling stress coerces cigarette seedling is tending towards normal and alleviates the generation of suppression " early blossoming ", leaf tobacco production has a good application prospect.
Accompanying drawing explanation
Fig. 1 be tobacco plant Chilling stress response ratio comparatively;
A: here adjoining tree withers, B: the present invention's anti-cryogen process plant, C/D/E: adjoining tree blade face chilling injury symptom;
Fig. 2 coerces tobacco plant normal temperature restoration ecosystem epidemic situation comparison through process of the present invention and the experience Chilling stress without process of the present invention;
Fig. 3 coerces tobacco plant situation of buddingging through process of the present invention and the experience Chilling stress without process of the present invention to compare;
Fig. 4 coerces the tobacco plant situation of blooming through process of the present invention with the experience Chilling stress without process of the present invention to compare;
Fig. 5 is the land for growing field crops pilot scale of the present invention's anti-cryogen yielding ability, A: field experiment, B: sample analyzed by flue-cured tobacco.
Embodiment
The present invention modulates multiple cold-resistant foliage spraying agent by using the physiologically active Auto-regulator of variety classes and proportioning, and its effect of comparison by experiment, filter out the cold-resistant foliage spraying agent of optimal component and proportioning.Technical scheme of the present invention is further illustrated by following concrete embodiment, and unrestricted the present invention.
Embodiment 1
The present invention's experiment is carried out in the Central-South tobacco experiment station of Agricultural University Of Hunan's Chinese tobacco.Produce main breed K326, cloud and mist 87 and No. 3, Hunan cigarette as tested variety using Tobacco In Hunan, use tobacco seedling dedicated substrate, adopt the nursery of floating seedlings method; After emerging, 1 ~ 2 leaf period thinning, 3 ~ 4 leaf periods are transplanted potted plant, carry out rich water quality management according to tobacco seedling technology, are that 6 ~ 8 leaf period cigarette seedlings are for experiment to being bred as.
One, the comparison of physiologically active of plant Auto-regulator foliage-spray treatment effect
According to the correlation of physiologically active of plant Auto-regulator and tobacco seedling stress resistance, consult domestic and international data of literatures in a large number, select the physiologically active of plant material that spermidine (Spd), betain (BH), proline (Pro), polyethylene glycol (PEG), 5-ALA (5-ALA) etc. 14 kinds are in close relations with plant stress resistance, carry out the screening of tobacco seedling Heat pretreatment conditioning agent component of the present invention, and determined the suitable concentration (see table 1) of each component tobacco leaf surface treatment by preliminary experiment.
Table 114 kind of physiologically active Auto-regulator and working concentration thereof
1, the process of physiologically active of plant Auto-regulator is on the impact of tobacco seedling stress response
By after the Potted tobacco seedling of 6 ~ 8 leaf ages immigration phytotron normal temperature domestication 3d (25 DEG C of 5000lx, illumination 12h), 14 Plants physiologically active Auto-regulators are with respective suitable concentration foliage-spray process tobacco seedling (spraying clear water for contrast); After spraying process 1 day (24h), start low temperature and poor light according to adverse circumstance process 10 days (8 DEG C of 1000lx, illumination 12h).
1.1 cigarette seedling Chilling stress responses
Low temperature and poor light according to adverse circumstance process the 0th, 0.5,1,2,3,7d noon, observe cigarette seedling Chilling stress responsive status, records appraisal respectively process whole strain cigarette seedling and blade wilting degree, damage to plants caused by sudden drop in temperature symptom etc.Result shows: during Chilling stress process, and wilting in various degree appears in most of tobacco seedling of contrast, and the blade of part contrast seedling occurs that water stain spot or leaf margin are rearwardly curling; The leaf surface treatment of majority of plant physiologically active Auto-regulator significantly can alleviate the here degree of withering of seedling, and wherein chilling injury symptom does not appear in most of seedling of the leaf surface treatment such as spermidine (Spd), betain (BH), 5-ALA (5-ALA), proline (Pro), salicylic acid (SA) and ABA.
Tobacco plant upgrowth situation and situation of buddingging after 1.2 recovery normal temperature
(1) after 10d low temperature treatment terminates, potted plant cigarette seedling is placed in normal temperature normal management, observes tobacco seedling growth and situation of buddingging.Observe and find: in normal temperature initial stages of restoration, the cigarette seedling normal growth of most of physiologically active Auto-regulator process compared with the control recovers very fast, and after 15d, all cigarette seedlings of 3 kinds all restore normal growth; Later stage (25d) observes display, and the tobacco seedling growth of part physiologically active Auto-regulator process is more vigorous, growth potential is better than contrast.
(2) normal temperature restoration ecosystem 30,45, the condition survey of buddingging of cigarette seedling after 60d.Observe and find: there is obvious difference the tobacco plant that same kind, different physiologically active Auto-regulator spray process time of buddingging, the time of buddingging of betain, proline, 5-ALA, salicylic acid and ABA process plant all comparatively contrasts obvious " postponement ".
2, Chilling stress process cigarette seedling leaf electrical conductivity detects
Relative conductivity is an important physical signs of reflection biofilm system situation.When plant meets with environment stress, cell membrane suffers destruction, permeability increases, and exosmosing of electrolyte inside cell will cause plant tissue leaching liquor electrical conductivity to increase, and namely membrane permeability increases degree and stress resistance of plant negative correlation.
Low temperature and poor light according to process period the 0th, 2,4,6,8d noon, the relative conductivity of the 4th or 5 blades of 14 Plants physiologically active Auto-regulator process cigarette seedlings is detected, to verify that it affects cigarette seedling Chilling stress resistance further.Testing result shows: compared with the control, the process of majority of plant physiologically active Auto-regulator can alleviate the increase that Chilling stress coerces caused blade relative conductivity, especially alleviate successful with this 5 Plants physiologically active Auto-regulator of betain, 5-ALA, proline, salicylic acid and ABA, namely enhance the Chilling stress resistance of cigarette seedling.
Overall merit is participated in the experiment 3 kinds, Chilling stress response performance (the blade wilting degree of 14 Plants physiologically active Auto-regulator leaf surface treatment seedling, damage to plants caused by sudden drop in temperature symptom, blade electrical conductivity), the situation (table 2) of normal temperature restoration ecosystem and performance three aspect of buddingging, and consider its physicochemical property, in this 14 Plants physiologically active Auto-regulator process, betain (BH), proline (Pro), salicylic acid (SA), the humidification of process to tobacco seedling Heat pretreatment of 5-ALA (5-ALA) and abscisic acid (ABA) is obvious, its process seedling shows stronger low-temperature resistance, normal temperature restoration ecosystem is good.Accordingly, the main candidate composition that these five kinds of physiologically active Auto-regulators are considered as Resistance In Tobacco cryogen is carried out further research.
Table 214 kind of physiologically active Auto-regulator is on the overall merit of tobacco bred K326 seedling Chilling stress response impact
Note: the evaluation of tobacco seedling low temperature stress reaction:
Two, the screening of physiologically active of plant Auto-regulator compound prescription
1, basic situation is tested: the Candidate components confirmed according to single physiologically active of plant Auto-regulator Preliminary screening in early stage and concentration thereof, design physiologically active Auto-regulator compound prescription combines 8 (table 3), and foliage-spray process starts low temperature and poor light according to adverse circumstance process after 1 day.Tested variety is K326 and cloud 87, and incubation, the phytotron of experiment material are tamed, low temperature and poor light is identical with previous experiments according to adverse circumstance treatment conditions, observation item and evaluation criterion.Low temperature and poor light is according to during process, 0th, 0.5,1,2,3,5,7d noon, observe, record Chilling stress response (wilting degree, damage to plants caused by sudden drop in temperature symptom) of each compound prescription process cigarette seedling, 0th, 1,3, each process cigarette seedling leaf photosynthetic activity is measured in the 5d noon, 0th, 2,4,6, the fresh sample of each process tobacco leaf is gathered in the 8d noon, after liquid nitrogen frozen grinds,-80 DEG C of low temperature refrigerators are preserved for physico-chemical analysis, and experimental result is in table 4,5.
The 8 kinds of compound prescription combinations of table 3 physiologically active Auto-regulator and concentration thereof
Note: in table, eight combinations are respectively the various combination formula of betain (B), salicylic acid (S), ABA (Ab), 5-ALA (A), proline (P).
2, the result of compound prescription conditioning agent and analysis (table 4)
2.1 Chilling stress responses
2. low temperature response (the damaging to plants caused by sudden drop in temperature symptom, wilting degree etc. of cigarette seedling) during overall merit tested variety K326, cloud and mist 87, the process of different composite formula process cigarette seedling Chilling stress, combine in 8 compound prescriptions, 3., 6., the Chilling stress response that 8. processes cigarette seedling be obviously better than contrast.
2.2 photochemical vitality
Chilling stress process period two, the photosynthetic activity of tested variety cigarette seedling declined (data slightly) all to some extent, but compound prescription process can alleviate the decline degree of cigarette seedling photosynthetic activity, wherein less, other compound prescription process cigarette seedling of the amplitude that declines of its photosynthetic activity of cigarette seedling of 2., 3., 6. processing of compound prescription decline by a big margin or with contrast suitable.
2.3 normal temperature growths recover
10d low temperature treatment terminates, recover normal temperature growth condition.After normal temperature restoration ecosystem 15d, the cigarette seedling of all process of two kinds all restore normal growth, wherein the tobacco seedling growth that 2., 3., 6. processes of compound prescription more vigorous (being obviously better than contrast).
The process of generalized analysis 8 Plants physiologically active Auto-regulator compound prescription foliage-spray is on the impact of two kind seedling Chilling stress responses and the upgrowth situation between normal temperature convalescence of participating in the experiment, compound prescription general performance 2., 3., is 6. more excellent, can be used as candidate's compound prescription, verifies screening further to its physiologic effect.
Table 4 physiologically active Auto-regulator compound prescription is on the preliminary assessment of tested variety seedling Chilling stress response impact
In addition, as can be seen from the physiologic effect of above-mentioned 8 compound prescriptions, the combined effect containing proline (Pro) is not good enough; Therefore, " rejecting " proline in five kinds of Candidate components, betain (BH), salicylic acid (SA), abscisic acid (ABA) and 5-ALA (5-ALA) four kinds of components are used for screening compound prescription further as preferred component.
3, the process of four kinds of preferred plant physiologically active Auto-regulators is on the impact of tobacco stress physiology biochemical indicator
During plant experience environment stress, its elementary tactics is active accommodation own metabolism, strengthen single-minded Resistance Physiology metabolism, accumulation compatible solute, as increased the endogenous cellular Protective substances content such as soluble sugar, proline and improving the activity of enzyme in antioxidant system; Meanwhile, by compatible solute Scavenger of ROS in cell, alleviate oxidation directly injury or play the 26S Proteasome Structure and Function being used for Protein requirement of molecular chaperones, protective plant escapes injury.Therefore, the physiological and biochemical index that stress resistance is correlated with is the important evidence of the stress resistance evaluating plant.
Make further screening for the preferred ingredient resisting cryogen to determine, to 4 kinds of preferred component betains (BH), salicylic acid (SA), abscisic acid (ABA) and 5-ALA (5-ALA) leaf surface treatment K326 cigarette seedling (incubation of material is the same), low temperature and poor light according to process the 0th, 2,4,6,8d to sample and to carry out physiological and biochemical analysis detection noon; Testing index: chlorophyll, malonaldehyde, proline, soluble protein, soluble sugar, ascorbic acid, hydrogen peroxide, content of oxygen free radical.
Table 5 four kinds of physiologically active Auto-regulators are to the overall merit of K326 seedling stress physiology Index Influence
Note: change compared with the control: significantly increase ++ increase+quite ± reduce-significantly reduce--
Overall merit: be better than contrast √ be inferior to contrast ×
Systematic analysis technique betain, salicylic acid, the leaf surface treatment of abscisic acid and 5-ALA 4 kinds of physiologically active Auto-regulators is to tobacco seedling stress physiology biochemical (table 5, data are slightly) show: the process of this 4 Plants physiologically active Auto-regulator, proline in tobacco seedling can be made, soluble protein, the content of the compatible solutes such as soluble sugar increases, reduce the accumulation of MDA and active oxy group, alleviate cellulose, chlorophyllous oxidative damage, all there is facilitation (consistent with the experimental result that early stage, tobacco seedling Chilling stress responded) to the enhancing of tobacco seedling stress resistance, this experimental result confirms the preferred ingredient that this 4 Plants physiologically active Auto-regulator can be used as Resistance In Tobacco cryogen compound prescription further.
Embodiment 2
Four kinds of preferred ingredients embodiment 1 finally obtained obtain new formula in conjunction with other auxiliary elements.
(amount) is claimed to get betain 1.15g, salicylic acid 5.0mg, 5-ALA 0.6g, abscisic acid 3mg, glycerine 20ml, polysorbas20 (Tween20) 0.5ml, Vc0.50g, calcium chloride 0.6g and trace elements iron (Fe) 5mg, boron (B) 5mg, manganese (Mn) 0.5mg, zinc (Zn) 10mg, molybdenum (Mo) 0.05mg, for the preparation of tobacco seedling cold-resistant foliage spraying agent.
First component salicylic acid is dissolved in 10ml absolute ethyl alcohol, obtains solution I;
Component betain, 5-ALA, glycerine, Vc, calcium chloride and trace element are added in 1L water, constantly stir until fully dissolve acquisition solution II;
Component abscisic acid is dissolved in 10ml absolute ethyl alcohol, obtains solution III;
Finally solution I, III to be joined respectively in solution II and to add polysorbas20, constantly stirring and make it fully mix, obtained cold-resistant foliage spraying agent of the present invention.
Embodiment 3
Take betain 1.85g, salicylic acid 10mg, 5-ALA 1g, abscisic acid 4mg, all the other components, content and preparation steps are with embodiment 2.
Embodiment 4
Take betain 2.25g, salicylic acid 8mg, 5-ALA 0.8g, abscisic acid 5mg, all the other components, content and preparation steps are with embodiment 2.
In order to prove the effect of the anti-cryogen formula of the present invention further, proceed the verification experimental verification of embodiment 5 ~ 8.
Embodiment 5: anti-cryogen process prepared by the embodiment of the present invention 2 is on the impact of tobacco seedling Resistant physiological index
Tested 4 ~ July in 2013,4 ~ July in 2014, in Agricultural University Of Hunan, Central-South tobacco experiment station large-sized artificial climatic chamber carries out.Experimental cultivar is K326 and cloud 87, and experiment material is bred as with embodiment 1.The seedling that growth is vigorous, growing way is basically identical is selected to move into phytotron, normal temperature domestication 3d (22 ~ 23 DEG C, 5000lx, illumination 12h); Chilling stress process (8 DEG C, 1000lx, illumination 12h) is started after anti-cryogen foliage-spray 24h.
Setup Experiments: anti-cryogen group (processed group) prepared by embodiment 2 and fresh water spraying group (control group), often group sprays process 10 strain (drenching as degree with blade upper and lower surface), repeats 3 times; After spraying process 24h, start Chilling stress process 7 days.Chilling stress process terminates rear phytotron and recovers normal temperature condition, and process cigarette seedling was moved back into the management of outdoor self-sow environmental cultivation from phytotron after 3 days by normal temperature condition, the fertility condition of period observation investigation tobacco plant.
Within the 0th, 1,3,5 day during Chilling stress process, measure the electrical conductivity of mature leaf, and gather middle part climax leaves sample for physiological and biochemical index analysis.Main Testing index has chlorophyll content, soluble sugar content, free proline content, SOD enzyme activity, mda content (table 6).
As can be seen from Table 6: under same experience Chilling stress condition, the Resistant physiological index of anti-cryogen processed group tobacco seedling is obviously better than control group (variation tendency of two kinds of participating in the experiment is similar); The single-minded physiological metabolism describing anti-cryogen process plant strengthens, improves the amount of compatible solute, and active oxygen elimination ability is also enhanced, for the Heat pretreatment improving tobacco seedling provides physiological and biochemical basis simultaneously.
The anti-cryogen process of table 6 is on the impact meeting with Chilling stress tobacco plant physiological and biochemical index
Embodiment 6: anti-cryogen process prepared by embodiment 2 is on the impact meeting with Chilling stress tobacco plant photosynthetic activity
Tested 4 ~ July in 2013,4 ~ July in 2014, in Agricultural University Of Hunan, Central-South tobacco experiment station large-sized artificial climatic chamber carries out, and experiment material and incubation thereof, Setup Experiments, Chilling stress process, with embodiment 5, the strain of every experimental group 5, to repeat 3 times.The the 0th, 1,3,5 day during Chilling stress process afternoon, the photosynthetic activity of tobacco plant mature leaf during using the photosynthetic analyzer of Li-6400 to detect Chilling stress process, main Testing index is Net Photosynthetic Rate (Pn), stomatal conductance (Gs), intercellular CO
2concentration (Ci).
The anti-cryogen process of table 7 is on the impact meeting with Chilling stress tobacco plant photosynthetic activity
Temperature is one of principal element affecting photosynthesis of plant, bad environmental temperature (high temperature or low temperature) will produce obvious inhibitory action to its photochemical vitality, cause its photosynthetic capacity to reduce and bring out the photosynthetic low temperature Xanthophyll cycle of plant.The Net Photosynthetic Rate of plant leaf blade is one of leading indicator of Photosynthetic ability of mesophyll size; Stomatal conductance represents the stretching degree of pore, has influence on blade O
2, CO
2with the exchange of steam and external environment, thus affect photosynthesis; When some " condition " one timing, the reduction of Photosynthetic ability of mesophyll will cause intercellular CO
2increasing of concentration.
Photosynthetic activity testing result display (table 7): under the adverse environmental factor of same experience low temperature and poor light photograph, anti-cryogen processed group plant has higher photosynthetic activity, main photosynthetic activity index net photosynthesis activity, stomatal conductance and intercellular CO
2the change of concentration is all better than control group (variation tendency of two kinds of participating in the experiment is similar).Can infer: when meeting with Chilling stress, the photosynthetic capacity of anti-cryogen process tobacco plant can maintain higher level compared with the control, the harmful effect of giving birth to tobacco for alleviating cold in the late spring provides " material " basis, will be conducive to " and seedling stage ", the growing of " stretching the root phase " tobacco plant.
Embodiment 7: the anti-cryogen process prepared of embodiment 2 on meeting with the response of Chilling stress tobacco plant Chilling stress, normal temperature growth recovers, budding, the impact of flowering time
Tested 4 ~ 7,2015 year 4 ~ July in 2014, at Agricultural University Of Hunan's Chinese tobacco, Central-South tobacco experiment station carries out.Incubation, the experimental design of experiment material are identical with embodiment 5 with adverse circumstance process.The Chilling stress response performance wilting degree of whole strain and the blade (, damage to plants caused by sudden drop in temperature symptom etc.) of tobacco plant is observed during adverse circumstance process; Move to outdoor normal temperature condition cultivation management after Chilling stress Stress treatment, observe the fertility condition of tobacco plant: the growth recovery state (growth recovery speed) at normal temperature initial stage, number of days different in flowering stage of buddingging, effective blade number.
The anti-cryogen process of table 8 is on the impact meeting with the fertility of Chilling stress tobacco plant
As can be seen from during adverse circumstance process and the observed result (table 8) of normal temperature restoration ecosystem: same meet with the condition of low temperature and poor light photograph under, the Chilling stress response of anti-cryogen processed group plant is obviously better than contrasting (Fig. 1), normal temperature condition initial stages of restoration, the growth (growth recovery speed) of processed group plant is also significantly better than control group (Fig. 2); In the normal temperature growth later stage, control group plant buddings, blooms, and the postponement about a week (Fig. 3,4) of blooming of buddingging of processed group plant.Experimental result shows: anti-cryogen process can strengthen the Chilling stress resistance of tobacco plant, reduce " damage " of Chilling stress, the early blossoming and the tobacco that can alleviate or suppress low temperature stress to cause buddings.In addition, effective the number of sheets comparatively control group increase about 2 ~ 3 of processed group tobacco plant.
Embodiment 8: the anti-cryogen process that embodiment 2 obtains is on the impact of tobacco flue-cured tobacco main components index
Tested 3 ~ August in 2015, carried out the pilot scale of yielding ability land for growing field crops in Changsha Yan Qu, cigarette district, Chaling and cigarette district, Yongzhou, experimental cultivar is respectively No. 5, K326, cloud 87 and Hunan cigarette.Experimental design is divided into anti-cryogen treatment region and fresh water spraying check plot, each experimental group repeat 3 times, every experimental group area is 20 × 3m
2.Cigarette seedling field-transplanting sprayed anti-cryogen after 15 days, field planting management thereafter identical with common production (Fig. 5 A); The tobacco leaf picking phase wins middle part Mature Tobacco Leaves and makes flue-cured tobacco sample (Fig. 5 B), analyzes main Flue-cured tobacco Quality component target: total reducing sugar, reducing sugar, nicotine, total nitrogen, K
+, Cl
-, content of starch.
The process of table 9 cold-resistant foliage spraying agent is on the impact of tobacco flue-cured tobacco main components index content (%)
Analysis result (table 9) as can be seen from three middle pilot flue-cured tobacco sample main components: the anti-main quality index content of cryogen treatment region flue-cured tobacco sample and the substantially close of check plot, all within normal range (NR), no significant difference; Describe the anti-cryogen modulated by physiological activator of the present invention to have no adverse effects to flue-cured tobacco main components.In addition, reflect: according to pilot scale point tobacco grower compared with check plot, anti-cryogen treatment region tobacco seedling growth gesture is more surging, and middle and upper part blade is slightly large, many about 3 of leaves number, flowering time pusher about 5 days of buddingging.
The anti-cryogen adopting the embodiment of the present invention 3 and 4 to prepare carries out embodiment 5,6,7,8 testing authentication, also can reach anti-cold effect good equally.
Claims (10)
1. a Resistance In Tobacco cryogen, is characterized in that, is the solution comprising following density component: betain 1.15 ~ 2.30g/L, salicylic acid 5.0 ~ 10.0mg/L, abscisic acid 3.0mg ~ 5.0mg/L, 5-ALA 600 ~ 1000mg/L.
2. anti-cryogen as claimed in claim 1, is characterized in that, the component preferred concentration comprised in described anti-cryogen: betain 1.15g/L, salicylic acid: 5.0mg/L, abscisic acid: 3.0mg/L, 5-glycyl acetic acid: 600mg/L.
3. anti-cryogen as claimed in claim 1, is characterized in that, also containing glycerine, Vc, calcium salt and trace element.
4. anti-cryogen as claimed in claim 3, is characterized in that, the calcium salt in described anti-cryogen is the one in nitrate or calcium chloride, and the concentration of calcium ion is 600 ~ 1000mg/L; The concentration of the Vc in described anti-cryogen is 500 ~ 1000mg/L, the concentration of glycerine is 15 ~ 20ml/L; Trace elements iron 5 ~ 10mg/L, boron 5 ~ 10/Lmg, manganese 0.5 ~ 1.0mg/L, zinc 10 ~ 25mg/L, molybdenum 0.05 ~ 0.10mg/L is also comprised in often liter of anti-cryogen.
5. anti-cryogen according to claim 1, is characterized in that, comprises one or more in 0.5ml plant leaf blade surface active agent tween 20, Tween 80, triton x-100 in often liter of anti-cryogen.
6. the preparation method of the anti-cryogen described in claim 1 or 2, it is characterized in that, step comprises:
(1) component salicylic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution I;
(2) component betain, 5-ALA are added to the water, constantly stir until fully dissolve acquisition solution II;
(3) component abscisic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution III;
(4) solution I and III are joined in solution II, and add plant leaf surface surfactant, constantly stir and make it fully mix, obtained described anti-cryogen.
7. the preparation method of the anti-cryogen described in any one of Claims 1 to 5, it is characterized in that, step comprises:
(1) component salicylic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution I;
(2) component betain, 5-ALA, Vc, glycerine, calcium salt and trace element are added to the water, constantly stir until fully dissolve acquisition solution II;
(3) component abscisic acid is dissolved in appropriate absolute ethyl alcohol or 95% ethanol, obtains solution III;
(4) solution I and III are joined in solution II, and add plant leaf surface surfactant, constantly stir and make it fully mix, obtained described anti-cryogen.
8. the using method of the anti-cryogen described in any one of Claims 1 to 5, is characterized in that, described anti-cryogen is at cigarette seedling foliage-spray.
9. the using method of anti-cryogen according to claim 8, is characterized in that, the rain or shine sky of " also seedling stage ", " stretching the root phase " breeding time foliage-spray at dusk after 5 ~ 7 days or cigarette seedling field-transplanting before cigarette seedling goes out garden.
10. the using method of anti-cryogen according to claim 8, is characterized in that, after using described anti-cryogen 3 ~ 5 times to dilute, foliage-spray, the every mu of amount of spraying are 25 ~ 30L.
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