CN101773004A - Method for improving cold resistance of pelleted seed of tobacco - Google Patents
Method for improving cold resistance of pelleted seed of tobacco Download PDFInfo
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- CN101773004A CN101773004A CN201010104989A CN201010104989A CN101773004A CN 101773004 A CN101773004 A CN 101773004A CN 201010104989 A CN201010104989 A CN 201010104989A CN 201010104989 A CN201010104989 A CN 201010104989A CN 101773004 A CN101773004 A CN 101773004A
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Abstract
The invention discloses a method for improving the cold resistance of pelleted seed of tobacco, which comprises the steps of: soaking and sterlizing tobacco seeds with 1.0 percent of copper sulfate for 15min, washing with clear water, initiating in 0.01 to 0.1mmol/L of polyamine solution under the dark condition at 20 to 25 DEG C for 24 to 48h, and adopting a conventional pelleting process to prepare the pelleted seed of tobacco. The invention has simple operation and low cost. After the tobacco seed is initiated by the polyamine solution, under the low temperature, the vitality of the pelleted seed and the quality of the seedling are high, the seedlings germinate fast and come out evenly, and growth potential is strong; and the protective enzymatic activity in the tobacco seedling is high, the malonaldehyde(MDA) content is low, and the low temperature resistance is obviously improved. The invention has better application prospect.
Description
Technical field
The invention belongs to tobacco seed processing technical field, be specifically related to a kind of method that can improve the tobacco seed cold tolerance.
Background technology
The eighties in 20th century, floating seedlings is applied to tobacco seedling the earliest in the U.S., and obtains developing rapidly and popularize in states such as the U.S., Canada, Japan immediately.Since promoting the floating seedlings technology on the tobacco, this technology fast development was popularized rapidly from beginning in 1996 in China, and national tobacco floating seed rearing popularity rate reached more than 70% in 2006.At present, tobacco floating seed rearing popularity rate in Yunnan Province's reaches more than 90%.
For ease of sowing, tobacco floating seed rearing generally adopts conventional coat pelleted seeds, to the resistivity of low temperature a little less than.Because big cigarette district temperature during floating seedlings of China is lower, and often meet bad weathers such as sleet is freezing, cause temperature of fry nursing pool lower, even can cause larval rearing water to freeze, the germination rate of coat pelleted seeds is reduced, emerge slowly, it is irregular to emerge, and a little less than the growing way, does not even emerge, have a strong impact on normal leaf tobacco production, bring the tremendous economic loss for country and tobacco grower.Therefore, how to improve the tobacco seed-coating pelleted seed resistance of low temperature is just become the technical problem that needs to be resolved hurrily on the leaf tobacco production.
Summary of the invention
The objective of the invention is at the deficiencies in the prior art, a kind of method that can effectively improve cold resistance of pelleted seed of tobacco is provided.
Purpose of the present invention is achieved by the following technical programs.
Except as otherwise noted, percentage of the present invention is percetage by weight.
A kind of method that improves cold resistance of pelleted seed of tobacco may further comprise the steps:
1, sterilization: tobacco seed uses 1.0% copper sulphate soaking disinfection 15min, and is clean with flushing with clean water, standby after taking out;
2, cause processing: will immerse through the tobacco seed of sterilization in the polyamines solution of 0.01～0.1mmol/L, the weight ratio of seed and polyamines solution is 1: 9, cause 24～48h down in 20 ℃～25 ℃ dark conditions, taking out seed cleans with clear water, at room temperature Hui Gan is to water content 4.0～7.0%, and is standby;
3, dressing processing: will cause the tobacco seed dressing Pelleting processing technology routinely after the processing, and promptly after dressing, nucleation, Pelleting, typing, polishing, dyeing and the drying, make the tobacco seed-coating pelleted seed.
Described polyamines solution is a kind of in putrescine (Put), spermidine (Spd) and spermine (Spm) solution.
The present invention is easy and simple to handle, and is with low cost.After naked kind of polyamines through 0.01～0.1mmol/L of tobacco causes 24～48h, under low temperature stress, the vigor of its coat pelleted seeds and seedling quality height, it is fast to germinate, and it is neat to emerge, and growth potential is strong; Protective enzyme in the tobacco seedling is active high, and malonaldehyde (MDA) content is low, and the resistance of low temperature is significantly strengthened.Have a good application prospect.
Embodiment
By specific embodiment given below, can further clearly understand the present invention.But they are not limitation of the invention.
Embodiment 1
Getting the tobacco seed 1.0kg (water content 5%) that quality reaches one-level breeding standard, is 1.0% copper sulphate soaking disinfection 15min with concentration.It is clean with flushing with clean water, standby to take out seed.Tobacco seed after the sterilization is put into the Put solution that 9.0kg concentration is 0.01mmol/L, place 20 ℃ of dark conditions under and to cause 48h, it is clean with clear water to take out seed, and at room temperature Hui Gan is to water content 4.0%, and is standby.To cause seeds treated dressing Pelleting processing technology routinely, and promptly after dressing, nucleation, Pelleting, typing, polishing, dyeing and the drying, make the tobacco seed-coating pelleted seed, the bag envelope is broadcast fully.
Embodiment 2
Repeat embodiment 1, following difference is arranged: the tobacco seed after will sterilizing is put into the Put solution that concentration is 0.1mmol/L, places to cause 24h under 25 ℃ of dark conditions, takes out seed and cleans with clear water, and at room temperature Hui Gan is to water content 7.0%.
Embodiment 3～4
Repeat embodiment 1～2, following difference is arranged: adopt Spd solution to cause.
Embodiment 5～6
Repeat embodiment 1～2, following difference is arranged: adopt Spm solution to cause.
Application Example 1
Getting the big gold dollar of 600g safflower (red big) seed (water content 5%), is 1.0% hydrogen peroxide dipping sterilization 15min with concentration.It is clean with flushing with clean water, standby to take out seed.Tobacco seed after the sterilization is divided into 6 parts, putting into 0.9kg concentration respectively is the Put solution of 0.00mmol/L (water), 0.01mmol/L and 0.1mmol/L, place under 20 ℃ of dark conditions and cause 24h and 48h respectively, taking out seed cleans with clear water, at room temperature Hui Gan is to water content 4.0%, and is standby.Undischarged seed is contrast to sterilize 15min through 1.0% copper sulphate.
With materials such as GTG pulvis, bentonite, talcum powder is auxiliary material, to cause seeds treated and contrast, dressing Pelleting processing technology routinely is promptly after dressing, nucleation, Pelleting, typing, polishing, dyeing and the drying, make the tobacco seed-coating pelleted seed, the bag envelope is broadcast fully.
Place the culture dish that is lined with two-layer moistening filter paper with causing coat pelleted seeds and conventional coat pelleted seeds, 16d germinates in 11 ℃ of 12h/12h (illumination/dark) illumination box, write down chitting piece number (counting germination more than or equal to seed length) every day, germinate and finish back calculating germination rate, germination index and average germinating time with radicle length.Average germinating time (d) MGT=∑ (Gt * Tt)/∑ Gt, germination index (GI)=∑ (Gt/Dt), in the formula, Gt is the germinative number at different time, Tt is the germination number of days.Test 3 repetitions, 200 seeds of every repetition.
The result shows: cause with contrast and water and handle ratio, Put causes and has improved seed germination rate and germination index under the low temperature stress, has shortened average germinating time (table 1).0.01mmol/L
Put initiation 48h processing germination rate and germination index are higher, handle than the water initiation of identical initiation time and have improved 21.0% and 40.0% respectively, and comparison is shone and improved 22.0% and 39.7% respectively respectively; Cause with contrast and the water of identical initiation time and to handle ratio, average germinating time has shortened 1.47 and 1.75d respectively.
The variation of red big coat pelleted seeds germination rate, germination index and average germinating time under table 1 low temperature stress
Behind the germination 16d, height of seedling (overground part+underground part) and root that every repetition picked at random 10 strain seedling are measured seedling with ruler are long, are accurate to mm, get its mean value.Each handles picked at random 50 strain seedling, cleans the back and dries to constant weight under 80 ℃, and the dry weight with analytical balance weighing seedling is accurate to 0.1mg, gets its mean value.
The result shows: cause with contrast and water and handle ratio, Put causes and has improved the height of seedling of tobacco seedling under the low temperature stress, root is long and seedling dry weight (table 2).0.01mmol/L it is higher that Put causes height of seedling, root length and the seedling dry weight of 48h processing, handles than the water initiation of identical initiation time and improved 18.8%, 61.8% and 53.7% respectively; Comparison is according to having improved 46.2%, 52.8% and 55.3% respectively.
The height of seedling of red big seedling, root length and dry weight change under table 2 low temperature stress
Germinate and finish back (16d), go to low temperature (5 ℃) and coerce 4d, (25 ℃) recover growth 4d under the normal temperature then, superoxide dismutase (SOD), peroxidase (POD), catalase (CAT) and ascorbate peroxidase enzyme (APX) activity and malonaldehyde (MDA) content measuring (4dayL) after (0day) before the low temperature treatment, the low temperature treatment and recover the normal tobacco seedling in growth back (4dayR).
Enzyme liquid preparation: in 0day, 4dayL and three periods of 4dayR, get seedling 0.3g respectively, 1.5mL grinds homogenate in ice bath with the pH7.0 phosphate buffer, centrifugal 20min under the 14000rpm, and the preservation of supernatant low temperature is standby, is used to measure POD, CAT and APX activity.Get seedling 0.3g, 1.5mL grinds homogenate in ice bath with the pH7.8 phosphate buffer, centrifugal 20min under the 14000rpm, and supernatant low temperature is preserved standby, is used to measure the SOD activity.
Nitrate blue tetrazolium (NBT) method is adopted in the SOD determination of activity, and reaction system is a 3mL NBT reactant liquor, and 0.1mL enzyme liquid reacts 20min under 30 ℃ of conditions in illumination box.Measure the OD value down in the 560nm wavelength, being suppressed 50% with NBT is an active unit (U), and enzymic activity is represented with U/gFWmin.
Guaiacol method is adopted in the POD determination of activity.The phosphate buffer 2.9mL of pH7.0,0.05M guaiacol 1mL and 2% H
2O
21mL mixes back 37 ℃ of insulations, adds behind the 0.1mL enzyme liquid mixing colorimetric under 470nm, OD value of per minute record.Changing 0.1 with per minute A470 is 1 unit of enzyme activity (U).
The method of (1994) such as Trevor is pressed in the CAT determination of activity.The phosphate buffer 3.88mL of pH7.0 and 30% H
2O
20.02mL mix back 37 ℃ of insulations, add behind the 0.1mL enzyme liquid mixing colorimetric under 240nm, OD value of per minute record.Changing 0.1 with per minute A240 is 1 unit of enzyme activity (U).
The APX determination of activity is revised slightly with reference to the method for Nakano and Asada (1981).The phosphate buffer 2.7mL of pH7.0 and the H of 300mM
2O
20.1mL mix back 25 ℃ of insulations, add behind the 0.1mL enzyme liquid mixing colorimetric under 290nm, OD value of per minute record.Changing 1.0 with per minute A290 is 1 unit of enzyme activity (U).
MDA Determination on content:, represent with thiobarbituricacid (TBARs) reactant with reference to Zhang Xianzheng (1992) method.Get seedling 0.4g fresh weight, add pH7.8 phosphate buffer 8mL and grind, gained homogenate is centrifugal 20min under 10000 * g, and supernatant is standby 4 ℃ of preservations.When measuring MDA, get the 1.5mL stock solution, add 2.5mL 5%TBA-TCA (trichloroacetic acid) liquid, boiling water bath 15 minutes, cooling with the centrifugal 10min of 1800 * g, is got supernatant and is measured absorbance with UV-2450 type spectrophotometer under 532nm and 600nm rapidly.Calculate the content (n mol/g FW) of MDA, extinction coefficient is 0.155.
The result shows: with ratio before the low temperature, after normal temperature recovered growth 4d, the SOD activity then significantly increased (table 3) without the red big seedling (CK) that causes processing.With ratio before the low temperature, low temperature stress is after 4 days, and the POD in the contrast seedling body is active significantly to raise, and normal temperature recovers then do not have marked change after the growth.
With the ratio in each of contrast seedling, after water caused 24h, the SOD activity in each period red big seedling body did not all have marked change in period, had improved normal temperature and had recovered the POD activity (table 3) in the seedling body behind the growth 4d.The POD activity that water initiation 48h handled each period does not make significant difference, but has reduced the SOD activity in the preceding seedling body of low temperature.
0.01mmol/L after Put causes 24h, improved the interior SOD activity of red big each seedling body in period, the POD activity in each period then do not had obvious influence (table 3).0.01mmol/L Put causes the 48h processing and has improved the SOD activity that reaches normal temperature recovery back seedling behind the low temperature, and the active nothing of POD is influenced.Handle and improved behind the low temperature and normal temperature recovers the SOD activity of back seedling 0.1mmol/L Put causes 24h, improved before the low temperature and the POD activity of seedling behind the normal temperature.Handle the SOD activity that has improved seedling in each 0.1mmol/L Put causes 48h in period, improved the POD activity of seedling behind the normal temperature.On the whole, 0.1mmol/L Put initiation 24h and 48h treatment effect are better, to SOD and the effect of being significantly increased of POD activity.
Interior SOD of red big seedling body and POD activity change under table 3 low temperature stress
With ratio before the low temperature, behind low temperature stress 4d and normal temperature recovery growth 4d, reduced CAT activity in the seedling without the red big seedling (CK) that causes processing, the APX activity then all significantly increases (table 4) in these two periods.With the ratio in each of contrast seedling in period, after water causes 24h and 48h, improved behind the low temperature and the CAT activity in the seedling body behind the normal temperature, improved the APX activity in 3 periods.
0.01mmol/L Put has only improved CAT and the APX activity of seedling behind the normal temperature after causing 24h.0.01mmol/L Put cause 48h handle improved behind the low temperature with normal temperature after seedling CAT activity, improved 3 period seedling APX activity (table 4).Handle and improved the CAT activity of seedling behind the normal temperature 0.1mmol/L Put causes 24h, 0.1mmol/L Put causes 48h to be handled and has then improved behind the low temperature and the CAT activity of seedling behind the normal temperature, two handle all obviously improved 3 period seedling the APX activity.In general, CAT and the APX activity of seedling were higher after 0.01mmol/L Put initiation 48h and 0.1mmol/L Put initiation 48h handled.
Interior CAT of red big seedling body and APX activity change under table 4 low temperature stress
With ratio before the low temperature, behind low temperature stress and the normal temperature growth, increased the MDA content (table 5) in the contrast seedling body.With the ratio in each of contrast seedling, water causes 24h and 48h two to be handled the MDA content in the seedling body before the low temperature and behind the low temperature all not to be had obvious influence in period, and variable concentrations Put causes the MDA content of handling seedling before the low temperature does not have obvious influence yet.Water causes 24h and 48h two to be handled and has all reduced normal temperature and recover the MDA content of growth back seedling, and 0.01mmol/L Put and the initiation of two times of 0.1mmol/L Put are handled and all reduced behind the low temperature and the MDA content of seedling behind the normal temperature.
MDA content in the red big seedling body under table 5 low temperature stress
Show based on the above results, under low temperature stress, after the Put of 0.01～0.1mmol/L causes 24～48h, germination rate, germination index, seedling length, root length and the seedling dry weight of its coat pelleted seeds have been improved, handle than the water initiation of identical initiation time respectively and on average improved 5.7%, 10.9%, 10.9%, 43.4% and 45.3%, comparison is according on average having improved 10.6%, 16.2%, 36.6%, 46.9% and 55.4% respectively; Average germinating time is handled than the water initiation of identical initiation time and has on average been shortened 0.56d, and comparison is according to having shortened 0.46d.Improved after the low temperature treatment and recovered the activity of superoxide dismutase (SOD), catalase (CAT), peroxidase (POD) and ascorbate peroxidase enzyme (APX) protective enzyme in the seedling body of growth back, reduced the content of the interior malonaldehyde (MDA) of seedling body.Illustrate that the tobacco seed-coating pelleted seed that causes pre-treatment through Put has stronger anti-anti-ability than conventional coat pelleted seeds to low temperature, has promoted the sprouting and the growth of seedlings of tobacco seed-coating pelleted seed under the low temperature stress.
Claims (2)
1. method that improves cold resistance of pelleted seed of tobacco may further comprise the steps:
(1) sterilization: tobacco seed uses 1.0% copper sulphate soaking disinfection 15min, and is clean with flushing with clean water, standby after taking out;
(2) cause processing: will immerse through the tobacco seed of sterilization in the polyamines solution of 0.01～0.1mmol/L, the weight ratio of seed and polyamines solution is 1: 9, cause 24～48h down in 20 ℃～25 ℃ dark conditions, taking out seed cleans with clear water, at room temperature Hui Gan is to water content 4.0～7.0%, and is standby;
(3) dressing processing: will cause the tobacco seed dressing Pelleting processing technology routinely after the processing, and promptly after dressing, nucleation, Pelleting, typing, polishing, dyeing and the drying, make the tobacco seed-coating pelleted seed.
2. the method for raising cold resistance of pelleted seed of tobacco according to claim 1 is characterized in that: described polyamines solution is a kind of in putrescine, spermidine and the spermine solution.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415236A (en) * | 2011-08-19 | 2012-04-18 | 青岛中烟种子有限责任公司 | Complete nutrition formula germination accelerating method for tobacco seed |
| CN111567557A (en) * | 2020-06-30 | 2020-08-25 | 玉溪中烟种子有限责任公司 | Initiator for improving cold resistance of tobacco seeds and initiation method |
| CN111802016A (en) * | 2020-07-06 | 2020-10-23 | 湖南省烟草公司永州市公司 | Method for accelerating germination of tobacco seeds |
| CN111869366A (en) * | 2020-08-18 | 2020-11-03 | 罗杰 | Method for priming tobacco seeds for extended storage time |
| CN112834703A (en) * | 2021-01-04 | 2021-05-25 | 湖南省烟草公司永州市公司 | Method for detecting activity of tobacco coated seeds |
| CN113994974A (en) * | 2021-11-17 | 2022-02-01 | 湖南省烟草公司永州市公司 | Initiator for improving cold resistance of tobacco seeds, initiation method and application |
-
2010
- 2010-02-03 CN CN201010104989A patent/CN101773004A/en active Pending
Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102415236A (en) * | 2011-08-19 | 2012-04-18 | 青岛中烟种子有限责任公司 | Complete nutrition formula germination accelerating method for tobacco seed |
| CN111567557A (en) * | 2020-06-30 | 2020-08-25 | 玉溪中烟种子有限责任公司 | Initiator for improving cold resistance of tobacco seeds and initiation method |
| CN111802016A (en) * | 2020-07-06 | 2020-10-23 | 湖南省烟草公司永州市公司 | Method for accelerating germination of tobacco seeds |
| CN111869366A (en) * | 2020-08-18 | 2020-11-03 | 罗杰 | Method for priming tobacco seeds for extended storage time |
| CN111869366B (en) * | 2020-08-18 | 2021-10-26 | 黔西南州烟草公司兴义市分公司 | Method for priming tobacco seeds for extended storage time |
| CN112834703A (en) * | 2021-01-04 | 2021-05-25 | 湖南省烟草公司永州市公司 | Method for detecting activity of tobacco coated seeds |
| CN113994974A (en) * | 2021-11-17 | 2022-02-01 | 湖南省烟草公司永州市公司 | Initiator for improving cold resistance of tobacco seeds, initiation method and application |
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Open date: 20100714 |