CN102783344A - Method for cultivating and planting mulberries in cold area - Google Patents
Method for cultivating and planting mulberries in cold area Download PDFInfo
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Abstract
The invention discloses a method for cultivating and planting mulberries in a cold area. China is a country with the most of mulberry types and mulberry cultivation area in the world, and is also the oldest country for cultivating and planting mulberries. The method for cultivating and planting the mulberries in the cold area comprises the following six steps of: 1, cold pre-treatment; 2, determining relative electric conductivity; 3, determining activity of superoxide dismutase (SOD); 4, determining concentration of malondialdehyde (MDA); 5, determining mass concentration of chlorophyll; and 6, determining photosynthetic performance. The method is used for cultivating and planting the mulberries in the cold area.
Description
Technical field:
the present invention relates to a kind of cold ground mulberry cultivation kind method for planting.
Background technology:
the Northeast belongs to the continental monsoon climate in temperate zone; Have the advantage that southern each province can't compare aspect kind of the Sang Yangcan, but then, because mulberry tree belongs to happiness temperature, happiness sun, the wet perennial woody plant of happiness; Suitable growth is in warmer area; And the Northeast's weather severe cold, winter is very long, frost-free season is short, and usually be low temperature, strong wind, arid spring, is the crucial restraining factors of planting Sang Yangcan.
29 kinds of
China mulberry, silkworm variety certification committee authorization (identifications) mainly are fit to the warm area of weather, the Yangtze river basin plants, and only has 2 suitable north latitude the Northeasts below 45 ° of kind of " No. 4, lucky lake " and " select autumn No. " to cultivate.When surpassing 45 ° of north latitude, " No. 4, lucky lake " sharply raises with the freeze injury rate of " select autumn No. one " 2 kinds, respectively up to 85% and 79%, causes enormous economic loss to peasant's planting mulberry and breeding silkworm.Through making great efforts for many years, the Northeast also successfully cultivates the mulberry tree new varieties with certain cold performance and (is fit to north latitude 45 Plant in the areas to the south).Like imperial mulberry No., Yilan 16 mulberry treies of Heilongjiang Province, the lucky "Hur" mulberry in Jilin Province, the Liao Dynasty in Liaoning Province educates No. 8 etc.These mulberry tree breeds are along with the continuous expansion of plant time limit growth and planting area, and the good characteristic of kind is degenerated gradually.Some high and cold area (mainly refer to 45-50 northeastward ) freeze injury take place frequently (the freeze injury rate of above-mentioned kind is all above more than 50%); And above-mentioned kind is main with the mulberry type of growing directly from seeds, yield poorly (single plant yield all is lower than 2kg), and picking leaves is taken a lot of work; Cause the appearance in a large amount of low yields mulberry field, be difficult to adapt to the requirement that the development of modern silkworm is already developed.The chief species of the local masses' planting mulberry and breeding silkworm of northeast high and cold area are for being a good unimproved wild mulberry tree; Though these mulberry tree frost resistances are good; But output extremely low (single plant yield all is lower than 1.5kg), quality of mulberry leaf poor (crude protein content is lower than 20%); Breeding silkworms low in economic efficiency (every mu of income is less than 700 yuan), had a strong impact on help already benefit and peasant of above-mentioned regional silkworm and planted the enthusiasm of Sang Yangcan, also is the crucial restraining factors of the strategic Project Realization of restriction China's " Nan Sang moves northward ".
in the evolution of the Northeast silkworm industry, the mulberry tree that lacks cold resistance, output height and quality better (is fit to north latitude 45.-50。Plant in the area) and supporting high-yield cultivation technique present situation; Integrated use one is breeding and cultivation technique measure; Select suitable the Northeast, northeast high and cold area particularly is like the good mulberry tree breed of Heilongjiang Province's Boreal climate characteristics; Change few, the disparate development of the above-mentioned regional mulberry tree breed of China, kind is difficult to adapt to the requirement of silkworm development.On the basis of cultivating the high-quality mulberry tree new varieties that cold resistance is strong, output is high; The principal element that has a strong impact on proterties such as mulberry tree output from mulberry tree top clipping, planting density etc. is studied, and sums up to propose the get bumper crops cultivation technique mode of efficient science of the Northeast mulberry tree.Therefore from implementing country's " Nan Sang moves northward " engineering, increase the high and cold regional silkworm raiser's income in northeast, enlarging the aspect consideration that is a good peasant employment and improves the ecological environment; The development trend of China's mulberry tree breeding is exactly to cultivate the mulberry tree of cold resistance, output height and quality better, and proposes corresponding culture technique on this basis.
China plants Sang Yangcan country the earliest in the world, and planting Sang Yangcan is China's conventional industries for thousands of years.According to statistics, at 1949 of founding of New, national mulberry field area had only 19.84 ten thousand hectares, 3.09 ten thousand tons of cocoon yields.China's sericulture industry is after the complications of passing through more than 20 year develop, and up to 1970, China's mulberry field area reached 23.34 ten thousand hectares, and 12.15 ten thousand tons of cocoon yields have surpassed Japan first, occupy the first in the world.Hereafter, China just occupies world sericulture industry giant's throne always.Since reform and opening-up, the hayday of China's sericulture industry development is in the eighties of last century front and back nineties in latter stage.1994, China's cocoon yield reached 77.69 ten thousand tons, and there are 26 provinces and cities' development sericulture industries in the whole nation.Nineteen ninety-five, national mulberry field area is 116.7 ten thousand hectares, and cocoon yield still reaches 75.98 ten thousand tons.National mulberry field area reached 710,000 hm in 2005
2
, produce 58.9 ten thousand tons of silk cocoons, nearly 70,000 tons of year reeling's output, silk weaving product 5,300,000,000 m, about 70,000,000,000 yuan of annual value of production ranks first in the world.
China is mulberry tree kind and the maximum country of mulberry cultivation area in the world, also is mulberry cultivation country with the longest history.Mulberry tree all has distribution all over China, divide according to taxonomy, can be divided into 15 mulberry kinds and 4 mutation; Can be divided into 8 Da Qu according to the physical geography situation; From the cultivation scope, northeast is on the south Harbin, and the northwest reaches Guangdong, Guangxi in the south from south, the Inner Mongol to Xinjiang, Qinghai, Gansu, Shaanxi, to the east of Taiwan, and to Sichuan, Yunnan, and with the Changjiang river, the Zhujiang River, the cultivation of the Yellow River three large watersheds at most.Can breed silkworms 4～5 times the whole year in the Yangtze river basin, amount of cocoon crop accounts for more than 80% of the whole nation, wherein flourishing with Sichuan, Zhejiang, Jiangsu San Sheng especially.Can breed silkworms 7～8 times the whole year in Zhujiang River basin, concentrate on ground such as the Shuande in Guangdong Province, middle mountain, and amount of cocoon crop accounts for about 7% of the whole nation.The Huanghe valley can be supported 3～4 times the whole year, mainly concentrates on provinces such as Shandong, Shanxi, Shaanxi, Hebei, Henan, and amount of cocoon crop accounts for the whole nation 6%.
Summary of the invention:
the purpose of this invention is to provide a kind of can be at cold district normal growth, cold ground mulberry cultivation kind method for planting that growing way is good.
Above-mentioned purpose realizes through following technical scheme:
a kind of cold ground mulberry cultivation kind method for planting; This method is six parts, and first is cold to handle, and the second portion relative conductivity is measured; Third part hepatocuprein SOD determination of activity; The 4th part MDA MDA concentration determination, the 5th part chlorophyll mass concentration is measured, and the 6th part photosynthetic performance is measured.
Described cold ground mulberry cultivation kind method for planting, described relative conductivity is measured, the mensuration of cell leakage; Beaker and glass apparatus are scrubbed clean with washing agent before use; With MiLLi-Q ultra-pure water drip washing 3 times, use MiLLi-Q ultra-pure water balance 24h again after the flushing with clean water, dry for use; Prosperous gloomy vast mulberry tree is got the blade of same area respectively with contrast Yilan 16 mulberry treies, and is clean with the running water rinsing, with ultra-pure water rinsing 3 times, after filter paper blots surface moisture, punches at random with card punch; In beaker, add 30mL MiLLi-Q ultra-pure water, with the complete submergence of leaf disk to be measured wherein, drain machine with vacuum and handle about 40min, when leaf disk is sunken to beaker bottom, room temperature measuring electrical conductivity S1; Its boiling water is boiled 20min, to be cooled to room temperature, measure electrical conductivity S2; Calculate relative conductivity according to formula:
In the formula: S
1
: the electric conductivity value of measuring under the room temperature; S
2
: boil the whole electric conductivity value that the back is measured; S
0
: the blank electric conductivity value of water.
Described cold ground mulberry cultivation kind method for planting, described hepatocuprein SOD determination of activity is the hepatocuprein assay method both; Get about 0.15g vegetable material to be tried,, put into the 5mL plastic centrifuge tube with being ground into powder behind the liquid nitrogen flash freezer; In centrifuge tube, add 2.5 1mL/15 mol/L phosphate buffer pH 7.8; Sonicated 1min, ice bath 5min, the centrifugal 5min of 12000g; Products obtained therefrom is transferred in another 5mL plastic centrifuge tube, and it is to be measured to put into 4 ℃ of storages of refrigerator; Get 1mL enzyme liquid with after 15 times of the products obtained therefrom supernatant dilutions, add 3mL reactant liquor 0.013 mol/L Met, 63 mol/L nitroblue tetrazoliums, 1.3 * 10
-6
Mol/L vitamin b3,1 * 10
-4
Mol/L EDTA and 0.05mol/L phosphate buffer, pH 7.8 dilutions; Behind 30 ℃, the 6 grades illumination box internal reaction 10min; Measure the 560nm light absorption value immediately, usefulness not reactant liquor 1mL 1/15mol/L phosphate buffer, the 3mL reactant liquor of enzyme-added liquid compares, with the zeroing of the enzyme liquid after the dilution;
With the light absorption value substitution following formula that records, try to achieve the SOD enzymic activity; The active unit of SOD is Umg
-1
SOD is active
In the formula:
A
0
: the absorbance value of irradiation control tube;
A
S
: the absorbance value of sample cell;
V
T
: sample enzyme liquid cumulative volume (mL);
V
1
: sample enzyme liquid long-pending (mL) during mensuration;
T: the reaction time (min);
W
F
: the clean quality (mgg of vegetable material powder
-1
).
In the formula:
A
450
: the absorbance value under the 450nm wavelength;
A
532
: the absorbance value under the 532nm wavelength;
A
600
: the absorbance value under the 600nm wavelength.
Described cold ground mulberry cultivation kind method for planting, described chlorophyll mass concentration is measured; Get the fresh blade of about 0.1g, grind into powder under liquid nitrogen flash freezer changes in the 5mL centrifuge tube, adds the precooling acetone reaction 5min of 1mL 80%, and the centrifugal 5min of 12000g is transferred to the products obtained therefrom supernatant in another centrifuge tube; Deposition continues extracting 2 times with the precooling acetone of 1mL 80%, and deposition is white in color fully; The acetone soln of extracting is several times mixed, again with after 5 times of 80% the acetone diluted under 663nm and 645nm wavelength the mensuration absorbance value, make blank with 80% acetone;
Calculate the mass concentration of chlorophyll a and b with formula:
Wherein:
C
a
: the mass concentration (gL of chlorophyll a
-1
);
C
b
: the mass concentration (gL of chlorophyll b
-1
);
C
A+b
: the mass concentration (gL of chlorophyll a+b
-1
);
C
A/b
: chlorophyll a, b mass concentration ratio.
Described cold ground mulberry cultivation kind method for planting, described photosynthetic performance is measured; Net Photosynthetic Rate day the process curve determination, from 08:00 to 18:00, every 2h adopts the portable photosynthetic analyzer of LI-6400 to measure the Net Photosynthetic Rate of primary vane, every young plant wood selects the mature leaf (the 4th leaf) on top to measure; The mensuration of photosynthetic-photoresponse curve from 09:30 to 11:00, adopts the portable photosynthetic mensuration of LI-6400 type system to measure each photosynthetic-photoresponse value spaceship-carried and ground contrast nursery stock respectively; Chloroplast light induced fluorescence strength detection adopts LI-6400 photosynthesis measurement system and 6400-40 fluorescence leaf chamber, respectively the chloroplast fluorescence intensity of nursery stock; Measure the fixing fluorescence of minimum fluorescence under its dark adaptation
F
0
With the total fluorescence of maximum fluorescence
F
Max
, calculate the relative unit of variable fluorescence and variable fluorescence rate, variable fluorescence by following formula:
F
v
=
F
Max
-
F
0
, variable fluorescence rate=
F
v
/
F
Max
Data processing method, data adopt photosynthetic mensuration method of inspection, and adopt comprehensive study to carry out statistical analysis.
Beneficial effect:
1. carry out the t check behind cell leakage, MDA concentration, SOD activity and the measuring chlorophyll content of the present invention to prosperous gloomy vast mulberry tree and contrast Yilan 16 mulberry tree blades; Prosperous gloomy vast, the physiological trait biochemical with contrast of table 1 comparison shows that as a result: cell leakage of prosperous gloomy vast mulberry tree and MDA concentration are respectively 4.81% and 0.5347 μ moLL-1; For according to low by 99.17% and 116.06%, and two proterties compare difference and reach respectively significantly and utmost point significance level with contrast, explain cold and coerce down; Prosperous gloomy vast mulberry tree keeps guaranteeing effectively normal the passing through property of cell membrane; Its film phase transition temperature maybe be lower, and therefore, the MDA concentration that produces on the cell membrane is obviously lower;
The SOD of prosperous gloomy vast mulberry tree is active, chlorophyll a/10
-3
, chlorophyll b/10
-3,
Be respectively 9.5615 (Ug with chlorophyll a/b
-1
FWmin
-1
), 15.722 (Ug
-1
FWmin
-1
), 4.08 (mgg
-1
FW) and 3.8534, be higher than contrast 309.51%, 204.34%, 121.97% and 168.78%, the utmost point is significantly higher than contrast respectively.This is illustrated in cold coercing down; Prosperous gloomy vast mulberry tree can keep normal growth to grow necessary physiology, the normal operation of biochemical function; And normal physiological, the biochemical function of contrast receive heavy damage at this moment, so new varieties all obviously are superior to contrast in growth, growth and pass economical character, economic characters technique of expression under cold is coerced;
Table 1
Annotate: t
0.05
=2.262, t
0.01
=3.25.
2. the present invention are observed through the field to this kind, and finding has extremely anti-mulberry bacteriosis performance, sees from continuous 3 years investigation result: the type mulberry does not have the mulberry bacteriosis takes place, and explains that this kind has extremely strong anti-mulberry bacteriosis proterties.
tree-like development 3. of the present invention, growth potential is vigorous, and the clockwork spring number is many, and branch is long; The branch bronzing, hibernaculum is big, shape tree-like development the triangular in shape, scale covers a watt arrangement, and branch is close to accessory bud in the left and right sides; Leaf scar shape semicircular in shape, leaf scar is big, and depression is darker, and internode 4.8cm young sprout color is a taupe, and climax leaves are oval or the circle decomposite leaf is mixed gives birth to, and leaf margin saw shape is shallow at the bottom of the leaf, blade tip cusp-shaped, blade size 18.9cm x 13.5cm, the thick 0.35mm of leaf; The swollen bud phase of male flower is May 8, and taking off the bud phase is May 12, and the swallow mouth phase is May 15, and the yellow phase is September 30, and the leaf fall period is October 26.
4. the present invention coerces down cold, and cell leakage of prosperous gloomy vast mulberry tree and MDA concentration are poor in significant difference and extremely apparent with the contrast mulberry tree, are respectively 4.81% and 0.5347 μ moLL
-1
, be lower than contrast 99.17% and 116.06%; The SOD activity is 9.5615 Ug
-1
FWmin
-1
, and have utmost point significant difference between the contrast mulberry tree, exceed contrast 309.51%.This shows that prosperous gloomy vast mulberry tree cold resistance obviously is superior to contrast, and this is the important biochemical basis that its degeneration-resistant border ability of coercing is better than contrast.
5. the present invention coerces down cold, and the directly related factor of the photosynthetic efficiency of prosperous gloomy vast mulberry tree extremely significantly is superior to contrast, like chlorophyll a/10
-3
, chlorophyll b/10
-3
, chlorophyll a/b, fixedly fluorescence, total fluorescence and variable fluorescence average are respectively 9.5615 Ug
-1
FWmin
-1
, 15.722Ug
-1
FWmin
-1
, 4.08mgg
-1
FW, 3.8534,148.0,716.2 and 568.2 exceeds contrast 309.51%, 204.34%, 121.97%, 168.78%, 27.92%, 26.18% and 25.73% respectively; Show that " prosperous gloomy vast mulberry tree No. " photosynthesis significantly is superior to contrast, this also is one of its inherent physiological Foundations with high yield and quality; The anti-mulberry bacteriosis proterties ability that prosperous gloomy vast mulberry tree is extremely strong, disease-free basically.
6. the present invention provide job through the mulberry tree plantation for the surplus rural labor; Accelerated rural laborer's transfer; This product has been obtained remarkable effect aspect the water and soil loss checking winds and fixing drifting sand, reduce, and has established solid foundation for realizing local agricultural sustainable development.
Embodiment:
Embodiment 1:
a kind of cold ground mulberry cultivation kind method for planting; This method was six steps, and the first step is cold to be handled, and the second step relative conductivity is measured; The 3rd step hepatocuprein SOD determination of activity; The 4th step MDA MDA concentration determination, the 5th step chlorophyll mass concentration is measured, and the 6th step photosynthetic performance is measured.
In the formula: S
1
: the electric conductivity value of measuring under the room temperature; S
2
: boil the whole electric conductivity value that the back is measured; S
0
: the blank electric conductivity value of water.
Described cold ground mulberry cultivation kind method for planting, described hepatocuprein SOD determination of activity is the hepatocuprein assay method both; Get about 0.15g vegetable material to be tried,, put into the 5mL plastic centrifuge tube with being ground into powder behind the liquid nitrogen flash freezer; In centrifuge tube, add 2.5 1mL/15 mol/L phosphate buffer pH 7.8; Sonicated 1min, ice bath 5min, the centrifugal 5min of 12000g; Products obtained therefrom is transferred in another 5mL plastic centrifuge tube, and it is to be measured to put into 4 ℃ of storages of refrigerator; Get 1mL enzyme liquid with after 15 times of the products obtained therefrom supernatant dilutions, add 3mL reactant liquor 0.013 mol/L Met, 63 mol/L nitroblue tetrazoliums, 1.3 * 10
-6
Mol/L vitamin b3,1 * 10
-4
Mol/L EDTA and 0.05mol/L phosphate buffer, pH 7.8 dilutions; Behind 30 ℃, the 6 grades illumination box internal reaction 10min; Measure the 560nm light absorption value immediately, usefulness not reactant liquor 1mL 1/15mol/L phosphate buffer, the 3mL reactant liquor of enzyme-added liquid compares, with the zeroing of the enzyme liquid after the dilution;
With the light absorption value substitution following formula that records, try to achieve the SOD enzymic activity; The active unit of SOD is Umg
-1
SOD is active
In the formula:
A
0
: the absorbance value of irradiation control tube;
A
S
: the absorbance value of sample cell;
V
T
: sample enzyme liquid cumulative volume (mL);
V
1
: sample enzyme liquid long-pending (mL) during mensuration;
T: the reaction time (min);
W
F
: the clean quality (mgg of vegetable material powder
-1
).
In the formula:
A
450
: the absorbance value under the 450nm wavelength;
A
532
: the absorbance value under the 532nm wavelength;
A
600
: the absorbance value under the 600nm wavelength.
Described cold ground mulberry cultivation kind method for planting, described chlorophyll mass concentration is measured; Get the fresh blade of about 0.1g, grind into powder under liquid nitrogen flash freezer changes in the 5mL centrifuge tube, adds the precooling acetone reaction 5min of 1mL 80%, and the centrifugal 5min of 12000g is transferred to the products obtained therefrom supernatant in another centrifuge tube; Deposition continues extracting 2 times with the precooling acetone of 1mL 80%, and deposition is white in color fully; The acetone soln of extracting is several times mixed, again with after 5 times of 80% the acetone diluted under 663nm and 645nm wavelength the mensuration absorbance value, make blank with 80% acetone;
Calculate the mass concentration of chlorophyll a and b with formula:
Wherein:
C
a
: the mass concentration (gL of chlorophyll a
-1
);
C
b
: the mass concentration (gL of chlorophyll b
-1
);
C
A+b
: the mass concentration (gL of chlorophyll a+b
-1
);
C
A/b
: chlorophyll a, b mass concentration ratio.
Described cold ground mulberry cultivation kind method for planting, described photosynthetic performance is measured; Net Photosynthetic Rate day the process curve determination, from 08:00 to 18:00, every 2h adopts the portable photosynthetic analyzer of LI-6400 to measure the Net Photosynthetic Rate of primary vane, every young plant wood selects the mature leaf (the 4th leaf) on top to measure; The mensuration of photosynthetic-photoresponse curve from 09:30 to 11:00, adopts the portable photosynthetic mensuration of LI-6400 type system to measure each photosynthetic-photoresponse value spaceship-carried and ground contrast nursery stock respectively; Chloroplast light induced fluorescence strength detection adopts LI-6400 photosynthesis measurement system and 6400-40 fluorescence leaf chamber, respectively the chloroplast fluorescence intensity of nursery stock; Measure the fixing fluorescence of minimum fluorescence under its dark adaptation
F
0
With the total fluorescence of maximum fluorescence
F
Max
, calculate the relative unit of variable fluorescence and variable fluorescence rate, variable fluorescence by following formula:
F
v
=
F
Max
-
F
0
, variable fluorescence rate=
F
v
/
F
Max
Data processing method, data adopt photosynthetic mensuration method of inspection, and adopt comprehensive study to carry out statistical analysis.
Embodiment 2:
Embodiment 1 described cold ground mulberry cultivation kind method for planting carries out the t check after the prosperous gloomy vast mulberry tree and the blade Net Photosynthetic Rate diurnal variation of contrast Yilan 16 mulberry tree blades measured, the diurnal variation umolm of result prosperous gloomy vast and contrast Net Photosynthetic Rate
-2
S
-1
Table 2 shows: prosperous gloomy vast mulberry tree blade Net Photosynthetic Rate annual average is 12.4887umolm
-2
S
-1
, and contrast is merely 8.3625umolm
-2
S
-1
, the two differs 149.4%, and difference is extremely remarkable.
Table 2
| Constantly | Prosperous gloomy vast mulberry tree | The contrast mulberry tree |
| 8:00 | 13.832 | 8.107 |
| 10:00 | 15.430 | 9.814 |
| 12:00 | 16.146 | 11.046 |
| 14:00 | 11.887 | 9.927 |
| 16:00 | 11.656 | 9.609 |
| 18:00 | 5.981 | 1.672 |
What photosynthetic-photoresponse curve of
plant reflected is the Changing Pattern that photosynthetic rate changes along with intensity of illumination, and the mensuration of this curve has important meaning for the photosynthetic capacity of judging plant.Under the certain environment condition, the maximum photosynthesis rate of plant leaf blade has been represented the maximum photosynthetic capacity of blade, and apparent quantum utilization ratio has reflected the utilize situation of blade to luminous energy, especially to the ability of utilizing of the low light level.Carry out the t check after leaf photosynthesis-photoresponse effect mensuration to prosperous gloomy vast mulberry tree and contrast Yilan 16 mulberry tree blades; The prosperous gloomy diurnal variation table 3 vast and the contrast Net Photosynthetic Rate of result shows: prosperous gloomy vast mulberry tree under the light intensity of measuring and contrast between photosynthetic rate average respectively 10.25 and 7.16, exceed contrast 43.16%, difference is extremely remarkable.But under the different high lights, the difference amplitude varies in size, and is lower than 100 μ molm in light intensity -2 s -1 The time, prosperous gloomy vast mulberry tree and contrast between photosynthetic rate average respectively 0.7266 and 0.5966, difference is not remarkable; When light intensity reaches 200 μ molm -2 s -1 When above, prosperous gloomy vast mulberry tree and contrast between the photosynthetic rate average reach 12.48 and 8.65 respectively, exceed contrast 47.75%, difference is extremely remarkable.This shows that under the different light intensity, the mulberry tree photosynthetic rate is different, (100 μ molm under the low light level -2 s -1 ), the performance of photosynthesis characteristic between different cultivars is insufficient, therefore prosperous gloomy vast mulberry tree and the contrast mulberry tree between do not have significant difference.But when light intensity is increased to 200 μ molm -2 s -1 When above, the characteristic that the new varieties photosynthetic efficiency is high fully shows, and exceeds contrast 47.75%.
Table 3
| Light intensity/μ molm -2·s -1 | Prosperous gloomy vast mulberry tree | The contrast mulberry tree |
| 0 | -1.90 | -2.24 |
| 50 | 0.80 | 0.67 |
| 100 | 3.28 | 3.36 |
| 150 | 6.43 | 5.18 |
| 200 | 8.31 | 6.50 |
| 300 | 10.68 | 7.80 |
| 400 | 11.96 | 8.40 |
| 500 | 12.76 | 8.80 |
| 600 | 13.00 | 9.30 |
| 800 | 13.50 | 9.40 |
| 1000 | 13.90 | 9.41 |
| 1200 | 14.00 | 9.42 |
| 1400 | 14.10 | 9.46 |
| 1600 | 14.60 | 9.70 |
| 1800 | 14.60 | 9.70 |
| 2000 | 14.40 | 9.40 |
The t check is carried out after the prosperous gloomy vast mulberry tree and the leaf chlorophyll fluorescence kinetics of contrast Yilan 16 mulberry tree blades are measured in
; The variation comparison sheet 4 of prosperous gloomy vast mulberry tree of result and contrast mulberry tree chlorophyll fluorescence shows: have utmost point significant difference between prosperous gloomy vast mulberry tree chlorophyll fluorescence kinetics characteristic and contrast mulberry tree; Wherein fixedly fluorescence, total fluorescence and variable fluorescence average are respectively 148.0,716.2 and 568.2, and comparison is according to having improved 27.92%, 26.18% and 25.73% respectively; This explains that prosperous gloomy vast photosyntheticsystem activity is significantly higher than contrast, and this also is that its yield and quality obviously is superior to one of important inherent physiological mechanism that contrasts.
Table 4
Annotate: t
0.01
=3.25;
*
Expression reaches utmost point significance.
Embodiment 3:
embodiment 1 described cold ground mulberry cultivation kind method for planting; Suitable temperature is the necessary environmental condition of plant growing; Injury from low temperature is the huge a kind of natural calamity of loss in the whole world agricultural and forestry production, and the annual loss that causes crops because of injury from low temperature in the whole world is up to thousands of dollars.Under the low temperature stress, a large amount of accumulation of plant membrane lipid peroxidating product MDA (MDA) can cause membrane permeability to rise, and electrolyte exosmoses, and make conductivity value become big, cause the major injury of cell membrane system.And when plant was in the low temperature adverse circumstance, the balance of free-radical generating and removing was destroyed in the cell, and the accumulation of active oxygen directly causes chlorophyllous destruction, and especially the specificity of chlorophyll a is destroyed, and caused chlorophyll to decompose and the decline of chlorophyll a/b ratio.And the strong plant of cold resistance, under the low temperature stress, its cell membrane phase transition temperature is lower; Film fat is difficult for oxidated generation MDA; And owing to the film system integrity protects, thereby the photosynthetical system function does not receive the influence of low temperature, therefore can keep the normal physiological function of cell largely.
cold resistant mechanism of prosperous gloomy vast mulberry tree in order further to illustrate is necessary the Physiology and biochemistry situation of change of prosperous gloomy vast mulberry tree under freeze injury is coerced carried out system research.
Claims (7)
1. cold ground mulberry cultivation kind method for planting; It is characterized in that: this method is six parts, and first is cold to handle, and the second portion relative conductivity is measured; Third part hepatocuprein SOD determination of activity; The 4th part MDA MDA concentration determination, the 5th part chlorophyll mass concentration is measured, and the 6th part photosynthetic performance is measured.
2. cold ground according to claim 1 mulberry cultivation kind method for planting is characterized in that: described cold the processing; Prosperous gloomy vast mulberry tree and each 100 strain of 16 mulberry treies of contrast Yilan; Test material is placed in the brooder, when outdoor temperature drops between 0～15 ℃, material placed carry out low temperature treatment in the booth; When temperature was handled 1 day-4 days for 3～15 ℃; Each plant is chosen 3 sites, averages, and is defined as the chlorophyll content of each transformed plant; When its Net Photosynthetic Rate was surveyed in 5～15 ℃ of processing of temperature in 1 day-6 days; Drew materials in first day and measure its MDA MDA content and relative conductivity, be designated as the base value of each physical signs, handle for 2～15 ℃ to draw materials in 7 days when temperature and survey its MDA MDA content and relative conductivity, be designated as the value of coercing; Below each material of mensuration of each proterties respectively measure 10 strains, every strain duplicate measurements 3 times;
3. cold ground according to claim 1 and 2 mulberry cultivation kind method for planting; It is characterized in that: described relative conductivity is measured; The mensuration of cell leakage scrubs clean beaker and glass apparatus before use with washing agent, after the flushing with clean water with MiLLi-Q ultra-pure water drip washing 3 times; Use MiLLi-Q ultra-pure water balance 24 hours again, dry for use; Prosperous gloomy vast mulberry tree is got the blade of same area respectively with contrast Yilan 16 mulberry treies, and is clean with the running water rinsing, with ultra-pure water rinsing 3 times, after filter paper blots surface moisture, punches at random with card punch; In beaker, add 30mL MiLLi-Q ultra-pure water, with the complete submergence of leaf disk to be measured wherein, drain machine with vacuum and handle about 40min, when leaf disk is sunken to beaker bottom, room temperature measuring electrical conductivity S1; Its boiling water is boiled 20min, to be cooled to room temperature, measure electrical conductivity S2; Calculate relative conductivity according to formula:
In the formula: S
1: the electric conductivity value of measuring under the room temperature; S
2: boil the whole electric conductivity value that the back is measured; S
0: the blank electric conductivity value of water.
4. according to claim 1 or 2 or 3 described cold ground mulberry cultivation kind method for planting, it is characterized in that: described hepatocuprein SOD activity determination method; Get about 0.15g vegetable material to be tried,, put into the 5mL plastic centrifuge tube with being ground into powder behind the liquid nitrogen flash freezer; In centrifuge tube, add 2.5 1mL/15 mol/L phosphate buffer pH 7.8; Sonicated 1min, ice bath 5min, the centrifugal 5min of 12000g; Products obtained therefrom is transferred in another 5mL plastic centrifuge tube, and it is to be measured to put into 4 ℃ of storages of refrigerator; Get 1mL enzyme liquid with after 15 times of the products obtained therefrom supernatant dilutions, add 3mL reactant liquor 0.013 mol/L Met, 63 mol/L nitroblue tetrazoliums, 1.3 * 10
-6Mol/L vitamin b3,1 * 10
-4Mol/L EDTA and 0.05mol/L phosphate buffer, pH 7.8 dilutions; Behind 30 ℃, the 6 grades illumination box internal reaction 10min; Measure the 560nm light absorption value immediately, usefulness not reactant liquor 1mL 1/15mol/L phosphate buffer, the 3mL reactant liquor of enzyme-added liquid compares, with the zeroing of the enzyme liquid after the dilution;
With the light absorption value substitution following formula that records, try to achieve the SOD enzymic activity; The active unit of SOD is Umg
-1, SOD is active
In the formula:
A 0: the absorbance value of irradiation control tube;
A S: the absorbance value of sample cell;
V T: sample enzyme liquid cumulative volume (mL);
V 1: sample enzyme liquid long-pending (mL) during mensuration;
T: the reaction time (min);
W F: the clean quality (mgg of vegetable material powder
-1)
5. according to claim 1 or 2 or 3 or 4 described cold ground mulberry cultivation kind method for planting, it is characterized in that: described MDA MDA concentration determination, get the 1.0g blade with liquid nitrogen flash freezer after; Be ground into powder, get and put into the 5mL plastic centrifuge tube in right amount, add 3mL10%TCA; Ice bath 5min, the centrifugal 10min of 12000g draws products obtained therefrom supernatant 2mL; Contrast adds 2mL distilled water; Add 2mL 0.6%TBA solution, homomixture reacts 15min in boiling water bath, and the dullness of this reactant liquor under 532nm or 600nm or 450nm wavelength measured in the cooling back; In the formula:
A 450: the absorbance value under the 450nm wavelength;
A 532: the absorbance value under the 532nm wavelength;
A 600: the absorbance value under the 600nm wavelength.
6. according to claim 1 or 2 or 3 or 4 or 5 described cold ground mulberry cultivation kind method for planting, it is characterized in that: described chlorophyll mass concentration is measured; Get the fresh blade of about 0.1g, grind into powder under liquid nitrogen flash freezer changes in the 5mL centrifuge tube, adds the precooling acetone reaction 5min of 1mL 80%, and the centrifugal 5min of 12000g is transferred to the products obtained therefrom supernatant in another centrifuge tube; Deposition continues extracting 2 times with the precooling acetone of 1mL 80%, and deposition is white in color fully; The acetone soln of extracting is several times mixed, again with after 5 times of 80% the acetone diluted under 663nm and 645nm wavelength the mensuration absorbance value, make blank with 80% acetone;
Calculate the mass concentration of chlorophyll a and b with formula:
Wherein:
C a: the mass concentration (gL of chlorophyll a
-1);
C b: the mass concentration (gL of chlorophyll b
-1);
C A+b: the mass concentration (gL of chlorophyll a+b
-1);
C A/b: chlorophyll a, b mass concentration ratio.
7. according to claim 1 or 2 or 3 or 4 or 5 or 6 described cold ground mulberry cultivation kind method for planting, it is characterized in that: described photosynthetic performance is measured; Net Photosynthetic Rate day the process curve determination, from 08:00 to 18:00, every 2h adopts the portable photosynthetic analyzer of LI-6400 to measure the Net Photosynthetic Rate of primary vane, every young plant wood selects the mature leaf (the 4th leaf) on top to measure; The mensuration of photosynthetic-photoresponse curve from 09:30 to 11:00, adopts the portable photosynthetic mensuration of LI-6400 type system to measure each photosynthetic-photoresponse value spaceship-carried and ground contrast nursery stock respectively; Chloroplast light induced fluorescence strength detection adopts LI-6400 photosynthesis measurement system and 6400-40 fluorescence leaf chamber, respectively the chloroplast fluorescence intensity of nursery stock; Measure the fixing fluorescence of minimum fluorescence under its dark adaptation
F 0With the total fluorescence of maximum fluorescence
F Max, calculate the relative unit of variable fluorescence and variable fluorescence rate, variable fluorescence by following formula:
F v=
F Max-
F 0, variable fluorescence rate=
F v/
F MaxData processing method, data adopt photosynthetic mensuration method of inspection, and adopt comprehensive study to carry out statistical analysis.
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| CN110706123A (en) * | 2019-11-19 | 2020-01-17 | 大连大学 | Screening method for identification index of higher soil pH value stress resistance of cowberry |
| CN112735511A (en) * | 2021-01-13 | 2021-04-30 | 西北农林科技大学 | Cold damage cucumber PSII potential activity prediction method based on QGA-SVR |
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