CN1623371A - Method for selecting tomato seed for resisting multi-disease and resurvey - Google Patents
Method for selecting tomato seed for resisting multi-disease and resurvey Download PDFInfo
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Abstract
A dicriminating technique for culturing the tomato varieties resisting multiple diseases and stress features that the TMV and/or leafmold and/or blight and/or meloidogyne are artificially inoculated to the plant of tomato respectively and the polyresistance to said diseases is then discriminated. The stress-resistance discrimination includes the weak light tolerance discrimination and low temp tolerance discrimination.
Description
(1) technical field
What the present invention relates to is a kind of breeding technique method of variety of crops, specifically a kind of breeding technique method of tomato variety.
(2) background technology
Tomato nutrient is abundant, and wide adaptability is of many uses, not only can be used as vegetable and fruit, and can be used as various fabricated products, also has health care, prevents and treats the effect of multiple disease.
Therefore tomato is one of main culturing vegetable in the world.Tomato also plays a part very importantly in China's vegetable supply, occupies an important position in the structural readjustment of rural industry.Along with the greenhouse by solar heat development, China's protection ground vegetable growing area constantly enlarges, and reaches 136.9 ten thousand hectares, and the plantation area of tomato accounts for about 40% in the protection ground.Tomato is again the main exit product that China develops export-oriented agricultural simultaneously, finds a good sale in countries such as Russia.Particularly after the China's entry into the WTO, no matter the outlet of vegetables still is that price has more intense competitive advantage from cost.
At present, many kinds of countries such as Holland and Israel are successively squeezed into China, and these kind plant strain growth gesture are strong, and it is good to continue the ability of bearing fruit, fruit is even, and pulp is hard, and deformity is few with dehiscent fruit, shelf life is long, and anti-multiple diseases has produced huge impact to the tomato market of China.
No matter tomato in China breeding 20 years is in the past still making great progress on breed breeding on the breeding technique, especially obviously dwindles with the international most advanced level gap aspect breeding for disease resistance.But the tomato in China breeding level is still very low.Show that mainly the degree of orientation of breeding is lower, the randomness of improved variety characteristic is big, and similarity factor is higher between kind.The average per unit area yield of tomato still is lower than world standard.The average yield of Holland's greenhouse type tomato long season cultivation is 50kg/m
2, the highest is 65kg/m
2, China lacks the tomato specific breed of storage tolerance, the suitable greenhouse culture of anti-adverse circumstance (tolerance to low temperature and weak light or heat-resisting).The tomato variety seed of China's export trade adopts the import seed of countries such as Holland, Israel mostly, costs an arm and a leg, and tomato is ten thousand yuan of 8-10 for every kilogram, is 200 times of tomato in China seed prices.Therefore seed selection is fit to greenhouse long season cultivation, and storage tolerance, anti-multiple diseases, tolerance to low temperature and weak light or heat-resisting tomato new varieties are task of top priority, and the kind that substitutes countries such as Israel, Holland, France is our target.Improve output and fruit storage tolerance and the long shelf life of tomato emphatically, improve exterior quality simultaneously, requirement is no malformed fruit, no-dehiscent fruit, and painted good, areola is tiny, even, the smooth rounding of fruit size, good, the mean fruit weight 160-200 gram of commodity.Except basic material lacked, its main cause was:
In breeding process, lack option program science, standard and effective choice, evaluation method and means.If this problem can not solve, the tomato breeding of China just is difficult to adapt to the needs of 21 century vegetables production development.Therefore, carry out disease-resistant, the degeneration-resistant authenticate technology of tomato and study, set up option program and technical system effectively reliably, improve constantly the level of tomato in China directive breeding.Make it to have the ability to cultivate the specific breed that is adapted to various planting types, various consumption demands.
(3) summary of the invention
The objective of the invention is to disease-resistant for cultivating, degeneration-resistant, suitable protection ground and outdoor cropping tomato variety anti-multiple diseases of tomato and breeding for stress tolerance authenticate technology method are provided.
The object of the present invention is achieved like this:
Comprise tomato carried out anti-multiple diseases authenticate technology and degeneration-resistant authenticate technology method that wherein anti-multiple diseases evaluation comprises carries out (1) TMV, leaf mold, the evaluation of fusarium wilt artificial inoculation on seedling multiresistance respectively; (2) TMV, leaf mold, root knot nematode disease artificial inoculation on seedling multiresistance are identified, (3) leaf mold, fusarium wilt, root knot nematode disease artificial inoculation on seedling multiresistance are identified; (4) TMV, fusarium wilt, root knot nematode disease artificial inoculation on seedling multiresistance artificial infection multiresistance are identified; (5) TMV, leaf mold, verticillium wilt artificial inoculation on seedling multiresistance are identified; (6) TMV, leaf mold, fusarium wilt, root-knot nematode artificial inoculation on seedling multiresistance are identified.
The various disease state of an illness investigation grade scales that anti-multiple diseases is identified are:
(1) leaf mold state of an illness investigation grade scale individual plant state of an illness grade scale:
0 grade: asymptomatic;
1 grade: the inoculation leaf chlorisis occurs to yellow scab;
3 grades: produce a small amount of white hypha, no spore on the inoculation leaf disease spot;
5 grades: produce mycelia and spore on the inoculation leaf disease spot;
7 grades: produce a large amount of mycelia and a large amount of spores on the inoculation leaf disease spot, and mycelia develops into upper leaf, but do not have spore;
9 grades: producing on the inoculation leaf disease spot has a large amount of spores, and spore also appears in upper leaf;
The disease-resistant grade scale of colony:
Immunity (I): do not infect, disease index is 0
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜15
Anti-disease (T): 15＜disease index＜30
Susceptible (S): disease index＞30
(2) the root knot nematode disease sentiment grade standard of checking the mark:
Individual plant state of an illness grade scale:
0 grade: unrooted tie lines worm and pieces of an egg
1 grade: 1-2 root knot and pieces of an egg
2 grades: 3-10 root knot and pieces of an egg
3 grades: 11-30 root knot and pieces of an egg
4 grades: 31-100 root knot and pieces of an egg
5 grades: 100 above root knots and pieces of an egg
The disease index computational methods
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is=0.
High anti-(HR): 0＜disease index＜1.0
Disease-resistant (R): 1.0＜disease index＜2.0
Anti-disease (T): 2.0＜disease index＜3.0
Susceptible (S): 3.0＜disease index＜4.0
High sense (HS): disease index＞4.0
(3) verticillium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: 1-2 sheet true leaf or two cotyledon flavescence, so that come off.
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging.
3 grades: complete stool is seriously wilted, so that withered.
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is 0.
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
In anti-(MR): 30＜disease index＜40
Anti-disease (T): 40＜disease index＜60
Susceptible (S): disease index＞60
(4) fusarium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: a slice true leaf or two cotyledon flavescence, so that come off.
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging.
3 grades: complete stool is seriously wilted, so that withered.
The disease-resistant grade scale of colony
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
Susceptible (S): disease index＞30
The compound criteria for classification that infects of TMV, CMV
The compound individual plant criteria for classification that infects of TMV, CMV
0 grade: asymptomatic
1 grade: leaf is not open and flat, light floral leaf
3 grades: the serious shrinkage of blade, heavy floral leaf
5 grades: heavy floral leaf, blade slightly is out of shape, the fern leaf
7 grades: heavy floral leaf, gross distortion, line leaf
9 grades: plant stops growing or is close to dead
(5) TMV, the disease-resistant grade scale of CMV colony
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜10
Anti-disease (T): 10＜disease index＜30
Susceptible (S): disease index＞30
Degeneration-resistant authenticate technology method comprises low light tolerance evaluation and low temperature resistant evaluation.
Wherein the low light tolerance authentication method is: select the strong and relatively poor varietal check of weak light resistance of known weak light resistance, under the normal illumination condition, use the nutritive cube seeding and seedling raising same period with to be identified material, each material plant is no less than 30 strains, after rough leaf was launched, each material was selected neatly, uniform seedling, was divided into two groups, every group 10 strain, one group still places under the normal illumination, and in contrast, another group is carried out shading treatment with black sunshade net.The all material of contrast and treatment region is arranged by completely random, contrast and processing should be arranged in the same facility, consistent to guarantee contrast with the temperature of handling, when the first inflorescence bud is obviously visible, investigate the plant height and the leaf area of every strain, according to investigation data calculate each material at leaf area, plant height under the shading with respect to the relative variation under the normal illumination, computing formula: relative variation=(numerical value of shading treatment-normal illumination numerical value) * 100%, carry out the leaf area of material to be identified, the relative variation of plant height and the contrast of low light tolerance relatively;
The low light tolerance ideal regression equation in seedling stage of low light tolerance authentication method is:
1. y=-0.7029x1-0.0550 x1: the thick ratio of plant height and stem relatively
2. y=-0.7173x2-0.0869 x2: chlorophyll b/a value relatively
3. y=-0.596x1-0.303x2-0.015 y: low light tolerance index
The phase low light tolerance regression equation of bearing fruit of blooming is:
4. y=-2.21x1+1.104 x1: the thick ratio of plant height and stem relatively
5. y=--0.524x2-0.1757 x2: chlorophyll b/a value relatively
Y: low light tolerance index
Low temperature resistant authentication method is: select the tomato that cold resistance is strong and cold resistance is weak to be contrast respectively, be seeded in the plastics nutritive cube same period with material to be identified, management routinely, when tomato long to the 4th true leaf launch, 5 true leaves are just current, every part of material is got 20 strains and is moved on under 2 ℃ of dark or the low light condition, after 48 hours, rank is damaged to plants caused by sudden drop in temperature in investigation; Select the strong and weak tomato of cold resistance to be contrast respectively, be seeded in the plastics nutritive cube same period with to be identified material and grow seedlings, field planting is in booth or greenhouse, manage routinely, remove first inflorescence, when first standard-sized sheet of using up treating second inflorescence put, get its pollen, on solid culture medium, observe down the sprouting situation in 8-12 ℃ of low temperature, medium making: with 80ml/L boric acid, 0.8% agar, after 10% sucrose medium was boiled mixing, adding one dripped on the spill slide and is made into solid culture medium, put into 8-12 ℃ growth case, after the equitemperature balance, the pollen that picks new collection with hair is broadcast in media surface, and it is capable to broadcast 2-4, examined under a microscope once every 15 minutes, measure pollen tube length and add up pollen germination rate and the length of measurement pollen tube with micrometer
In the low temperature resistant authentication method
(1) the low temperature identification of indicator of tomato seeds germination period is a seed germination index under 15 ℃ of conditions;
(2) do anti-cold direct indicator with the preceding two fringes relative fruit-setting rate really of seedling after the anti-cold exponential sum low temperature treatment, carry out correlation analysis simultaneously, think that relative conductivity, soluble sugar and soluble protein can be used as the indirect index of lower temperature resistance
Y=-0.6380x+2.6272 x: relative conductivity
Y=0.3397x+1.5130 x: relative protein content
(3) the bloom identification of indicator of the phase of bearing fruit is a pollen viability after low temperature treatment meiosis stage, seedling stage, and bud phase lower temperature resistance unanimity, and the florescence is inconsistent.
The classification that tomato is damaged to plants caused by sudden drop in temperature
0 grade: plant leaf is normal, is damaged to plants caused by sudden drop in temperature;
1 grade: only the minority blade edge has slight shrinkage to wilt;
2 grades: the following blade of half is wilted dead;
3 grades: blade more than half is wilted dead;
4 grades: plant is all dead.
According to the rank that damages to plants caused by sudden drop in temperature of investigation, calculate each material and damage to plants caused by sudden drop in temperature the anti-cold finger number of exponential sum.
Damage to plants caused by sudden drop in temperature index=∑ xa/ (n ∑ x)=(X1a1+X2a2)/nT
X1, X2 in the formula, X3 ... Xn represents the tomato numbers that damage to plants caused by sudden drop in temperature at different levels, a1, a2, a3 ... an represents respectively to damage to plants caused by sudden drop in temperature grade, and T is total book of investigation.Anti-cold finger number is the inverse that damages to plants caused by sudden drop in temperature index.
The present invention is concrete to propose in the diseases such as 1. anti-ToMV, leaf mold, fusarium wilt, root knot nematode disease, verticillium wilt disease artificial inoculation on seedling authentication method, inoculum density, inoculation seedling age, inoculation order more than three kinds, every kind of disease individual plant and colony's grade scale.2. the authentication method of tomato low light tolerance.Utilize relative plant height and stem thick than can be used as the identification of indicator of the tomato seedling low light level with relative chlorophyll b/a value, plant height, the thick ratio of stem and relatively soluble protein content can be as the identification of indicator of the setting phase low light tolerance material of blooming relatively.3. low temperature resistant authentication method, the relative percentage of fertile fruit that utilizes the preceding two fringes fruit of seedling after anti-cold finger mark and the low temperature treatment is as resisting cold direct indicator seedling stage, relative conductivity, soluble sugar and soluble protein can be used as the indirect index of lower temperature resistance, and the pollen viability power after the meiosis stage low temperature treatment can be used as the standard of perfection of the setting phase of blooming.And the grade scale that tomato is damaged to plants caused by sudden drop in temperature is proposed, calculating is damaged to plants caused by sudden drop in temperature the anti-cold finger of exponential sum and is counted pollen germination method under formula and the low temperature.Can select anti-multiple diseases by method of the present invention, the good tomato variety of tolerance to low temperature and weak light.
(4) specific embodiments
For example the present invention is done in more detail below and describes:
The present invention includes tomato is carried out anti-multiple diseases (disease more than three kinds in the main disease such as tomato ToMV, CMV, leaf mold, fusarium wilt, root knot nematode disease, verticillium wilt, early blight) artificial inoculation on seedling authenticate technology and degeneration-resistant (tolerance to low temperature and weak light) authenticate technology method.
1. tomato seedling multiresistance authentication method
(1) TMV, leaf mold, fusarium wilt sprout period testifying method
1. inoculation in proper order: TMV → leaf mold → fusarium wilt
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaves
3. inoculum density:
TMV:1 restrains phosphate buffer or the distilled water 10ml that the disease leaf adds 0.01Mph7.0, and inoculation back multiple connection in the 3rd day once.
Leaf mold: 10
6Individual spore/ml
Fusarium wilt: 10
7Individual spore/ml
4. inoculation method
TMV: rubbing manipulation
Leaf mold: spray-on process
Fusarium wilt: root-pouring method
5. investigate the date
TMV: inoculate back 20 days
Leaf mold: inoculate back 18 days
Fusarium wilt: inoculate back 21 days
(2) TMV, leaf mold, root knot nematode disease sprout period testifying method
1. inoculation in proper order: root-knot nematode → TMV → leaf mold
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaves
3. inoculum density:
TMV:1 restrains phosphate buffer or the distilled water 10ml that the disease leaf adds 0.01Mph7.0, and inoculation back multiple connection in the 3rd day once.
Leaf mold: 10
6Individual spore/ml
Root knot nematode disease: 5000/strain
4. inoculation method
TMV: rubbing manipulation
Leaf mold: spray-on process
Root knot nematode disease: root-pouring method
5. investigate the date
TMV: inoculate back 20 days
Leaf mold: inoculate back 18 days
Root knot nematode disease: inoculate back 45 days
(3) leaf mold, fusarium wilt, root knot nematode disease sprout period testifying method
1. inoculation in proper order: root-knot nematode → leaf mold → fusarium wilt
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaves
3. inoculum density:
Root knot nematode disease: 5000/strain
Leaf mold: 10
6Individual spore/ml
Fusarium wilt: 10
7Individual spore/ml
4. inoculation method
Fusarium wilt: root-pouring method
Leaf mold: spray-on process
Root knot nematode disease: root-pouring method
5. investigate the date
Fusarium wilt: inoculate back 21 days
Leaf mold: inoculate back 18 days
Root knot nematode disease: inoculate back 45 days
(4) TMV, fusarium wilt, root knot nematode disease sprout period testifying method
1. inoculation in proper order: root-knot nematode → TMV → fusarium wilt
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaves
3. inoculum density:
TMV:1 restrains phosphate buffer or the distilled water 10ml that the disease leaf adds 0.01Mph7.0, and inoculation back multiple connection in the 3rd day once
Root knot nematode disease: 5000/strain
Leaf mold: 10
6Individual spore/ml
Fusarium wilt: 10
7Individual spore/ml
4. inoculation method
TMV: rubbing manipulation
Root knot nematode disease: root-pouring method
Fusarium wilt: root-pouring method
5. investigate the date
Fusarium wilt: inoculate back 21 days
TMV: inoculate back 20 days
Root knot nematode disease: inoculate back 45 days
(5) TMV, leaf mold, verticillium wilt sprout period testifying method
1. inoculation in proper order: TMV → leaf mold → verticillium wilt
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaves
3. inoculum density:
TMV:1 restrains phosphate buffer or the distilled water 10ml that the disease leaf adds 0.01Mph7.0, and inoculation back multiple connection in the 3rd day once.
Leaf mold: 10
6Individual spore/ml
Verticillium wilt: 10
7Individual spore/ml
4. inoculation method
TMV: rubbing manipulation
Leaf mold: spray-on process
Verticillium wilt: root-pouring method
5. investigate the date
TMV: inoculate back 20 days
Leaf mold: inoculate back 18 days
Verticillium wilt: inoculate back 15 days
(6) TMV, leaf mold, fusarium wilt root-knot nematode sprout period testifying method
1. inoculation in proper order: root knot nematode disease → TMV → leaf mold → fusarium wilt
2. inoculate seedling age: 2 true leaf → 3 true leaf → 4 true leaf → 5 true leaves
3. inoculum density:
TMV:1 restrains phosphate buffer or the distilled water 10ml that the disease leaf adds 0.01Mph7.0, and inoculation back multiple connection in the 3rd day once.
Leaf mold: 10
6Individual spore/ml
Fusarium wilt: 10
7Individual spore/ml
Root knot nematode disease: 5000/strain
4. inoculation method
TMV: rubbing manipulation
Leaf mold: spray-on process
Fusarium wilt: root-pouring method
Root knot nematode disease: root-pouring method
5. investigate the date
TMV: inoculate back 20 days
Leaf mold: inoculate back 18 days
Fusarium wilt: inoculate back 21 days
Root knot nematode disease: inoculate back 45 days
Various disease state of an illness investigation grade scales
Leaf mold state of an illness investigation grade scale individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: the inoculation leaf chlorisis occurs to yellow scab
3 grades: produce a small amount of white hypha, no spore on the inoculation leaf disease spot.
5 grades: produce mycelia and spore on the inoculation leaf disease spot.
7 grades: produce a large amount of mycelia and a large amount of spores on the inoculation leaf disease spot, and mycelia develops into upper leaf, but do not have spore.
9 grades: producing on the inoculation leaf disease spot has a large amount of spores, and spore also appears in upper leaf.
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is 0.
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜15
Anti-disease (T): 15＜disease index＜30
Susceptible (S): disease index＞30
The root knot nematode disease sentiment grade standard of checking the mark
Individual plant state of an illness grade scale:
0 grade: unrooted tie lines worm and pieces of an egg
1 grade: 1-2 root knot and pieces of an egg
2 grades: 3-10 root knot and pieces of an egg
3 grades: 11-30 root knot and pieces of an egg
4 grades: 31-100 root knot and pieces of an egg
5 grades: 100 above root knots and pieces of an egg
The disease index computational methods
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is=0.
High anti-(HR): 0＜disease index＜1.0
Disease-resistant (R): 1.0＜disease index＜2.0
Anti-disease (T): 2.0＜disease index＜3.0
Susceptible (S): 3.0＜disease index＜4.0
High sense (HS): disease index＞4.0
Verticillium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: 1-2 sheet true leaf or two cotyledon flavescence, so that come off.
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging.
3 grades: complete stool is seriously wilted, so that withered.
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is 0.
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
In anti-(MR): 30＜disease index＜40
Anti-disease (T): 40＜disease index＜60
Susceptible (S): disease index＞60
Fusarium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: a slice true leaf or two cotyledon flavescence, so that come off.
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging.
3 grades: complete stool is seriously wilted, so that withered.
The disease-resistant grade scale of colony
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
Susceptible (S): disease index＞30
The compound criteria for classification that infects of TMV, CMV
The compound individual plant criteria for classification that infects of TMV, CMV
0 grade: asymptomatic
1 grade: leaf is not open and flat, light floral leaf
3 grades: the serious shrinkage of blade, heavy floral leaf
5 grades: heavy floral leaf, blade slightly is out of shape, the fern leaf
7 grades: heavy floral leaf, gross distortion, line leaf
9 grades: plant stops growing or is close to dead
TMV, the disease-resistant grade scale of CMV colony
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜10
Anti-disease (T): 10＜disease index＜30
Susceptible (S): disease index＞30
Three, tomato low light tolerance authentication method
1. authentication method
(1) tomato seedling and the setting phase weak light resistance separately and uncorrelated of blooming, plant height can be used as the identification of indicator of tomato seedling weak light resistance with the thick ratio of stem with relative chlorophyll b/a value relatively, plant height, the thick ratio of stem and relatively soluble protein content can be used as the identification of indicator of the setting phase low light tolerance material of blooming relatively, and it is as follows to set up regression equation:
Seedling stage low light tolerance ideal regression equation:
1. y=-0.7029x1-0.0550 x1: the thick ratio of plant height and stem relatively
2. y=-0.7173x2-0.0869 x2: chlorophyll b/a value relatively
3. y=-0.596x1-0.303x2-0.015 y: low light tolerance index
The setting phase low light tolerance regression equation of blooming is as follows:
4. y=-2.21x1+1.104 x1: the thick ratio of plant height and stem relatively
5. y=--0.524x2-0.1757 x2: chlorophyll b/a value relatively
Y: low light tolerance index
(2) select known weak light resistance strong (as caruso, in assorted 9, good powder etc.) and the varietal check of weak light resistance relatively poor (as middle vegetables 6 etc.), under the normal illumination condition, use the nutritive cube seeding and seedling raising same period with to be identified material.Each material (comprising control material) plant is no less than 30 strains.
After rough leaf was launched, each material was selected neatly, uniform seedling, was divided into two groups, every group 10 strain.One group still places under the normal illumination, and in contrast, another group is carried out shading treatment with black sunshade net. and all material of contrast and treatment region is arranged by completely random.Contrast and processing should be arranged in the same facility, and be consistent with the temperature of handling to guarantee contrast.
When the first inflorescence bud is obviously visible, investigate the plant height and the area of every strain.According to investigation data calculate each material at leaf area, plant height under the shading with respect to the relative variation under the normal illumination (%).
Computing formula: relative variation=(numerical value of shading treatment-normal illumination numerical value) * 100 (%), carry out the leaf area of material to be identified, the relative variation of plant height and the contrast of low light tolerance relatively.
Four, low temperature resistant authentication method
1. low temperature resistant authentication method
(1) the low temperature identification of indicator of tomato seeds germination period is a seed germination index under 15 ℃ of conditions.
(2) do anti-cold direct indicator with the preceding two fringes relative percentage of fertile fruit really of seedling after the anti-cold exponential sum low temperature treatment, carry out correlation analysis simultaneously, think that relative conductivity, soluble sugar and soluble protein can be used as the indirect index of lower temperature resistance
Y=-0.6380x+2.6272 x: relative conductivity
Y=0.3397x+1.5130 x: relative protein content
(3) the bloom identification of indicator of setting phase is a pollen viability after low temperature treatment meiosis stage, seedling stage, and bud phase lower temperature resistance unanimity, and the florescence is inconsistent.
Seedling stage the low temperature treatment method
Select the strong and weak tomato of cold resistance to be contrast respectively, in the material to be identified sowing same period and plastics nutritive cube, management routinely.Just current to the 4th true leaf expansion, 5 true leaves when tomato length, every part of material is got 20 strains and is moved on under 2 ℃ of dark or the low light condition.After 48 hours, rank is damaged to plants caused by sudden drop in temperature in investigation.The classification of damaging to plants caused by sudden drop in temperature can be with reference to following standard.
The classification that tomato is damaged to plants caused by sudden drop in temperature
0 grade: plant leaf is normal, is damaged to plants caused by sudden drop in temperature;
1 grade: only the minority blade edge has slight shrinkage to wilt;
2 grades: the following blade of half is wilted dead;
3 grades: blade more than half is wilted dead;
4 grades: plant is all dead.
According to the rank that damages to plants caused by sudden drop in temperature of investigation, calculate each material and damage to plants caused by sudden drop in temperature the anti-cold finger number of exponential sum.
Damage to plants caused by sudden drop in temperature index=∑ xa/ (n ∑ x)=(X1a1+X2a2)/nT
X1, X2 in the formula, X3 ... Xn represents the tomato numbers that damage to plants caused by sudden drop in temperature at different levels, a1, a2, a3 ... an represents respectively to damage to plants caused by sudden drop in temperature grade, and T is total book of investigation.Anti-cold finger number is the inverse that damages to plants caused by sudden drop in temperature index.The anti-cold finger number of material to be identified and cold-resistant and not cold-resistant contrast are compared, make corresponding evaluation.
3. pollen germination under the low temperature
Select the strong and weak tomato of cold resistance to be contrast respectively, and grow seedlings in the material to be identified sowing same period and the plastics nutritive cube, field planting manages in canopy or greenhouse routinely.
Remove first inflorescence.When first standard-sized sheet of using up treating second inflorescence put, get its pollen.On solid culture medium, observe the sprouting situation down in 8-12 ℃ of low temperature.Medium making: after 80ml/L boric acid, 0.8% agar, 10% sucrose medium boiled mixing, adding one drips on the spill slide and is made into solid culture medium, put into 8-12 ℃ growth case, after the equitemperature balance, the pollen that picks new collection with hair is broadcast in media surface, and it is capable to broadcast 2-4.Examined under a microscope once every 15 minutes, measure pollen tube length and add up pollen germination rate and the length of measurement pollen tube with micrometer.
Claims (5)
1, anti-multiple diseases of a kind of tomato and breeding for stress tolerance authenticate technology method, it is characterized in that: comprise tomato is carried out anti-multiple diseases authenticate technology and degeneration-resistant authenticate technology method that wherein anti-multiple diseases evaluation comprises carries out (1) TMV, leaf mold, the evaluation of fusarium wilt artificial inoculation on seedling multiresistance respectively; (2) TMV, leaf mold, root knot nematode disease artificial inoculation on seedling multiresistance are identified; (3) leaf mold, fusarium wilt, root knot nematode disease artificial inoculation on seedling multiresistance are identified; (4) TMV, fusarium wilt, root knot nematode disease artificial inoculation on seedling multiresistance are identified; (5) TMV, leaf mold, verticillium wilt artificial inoculation on seedling multiresistance are identified; (6) TMV, leaf mold, fusarium wilt, root knot nematode disease artificial inoculation on seedling multiresistance are identified; Degeneration-resistant authenticate technology method comprises low light tolerance evaluation and low temperature resistant evaluation, wherein the low light tolerance authentication method is: select the stronger and relatively poor kind of known weak light resistance to do contrast, under the normal illumination condition, use the nutritive cube seeding and seedling raising same period with to be identified material, each material plant is no less than 30 strains, after rough leaf is launched, each material is selected neat, uniform seedling, be divided into two groups, every group 10 strain, one group still places under the normal illumination, in contrast, another group is carried out shading treatment with black sunshade net, the all material of contrast and treatment region is arranged by completely random, contrast and processing should be arranged in the same facility, consistent to guarantee contrast with the temperature of handling, when the first inflorescence bud is obviously visible, investigate the plant height and the area of every strain, calculate each material leaf area under shading according to investigation data, plant height is with respect to the relative variation under the normal illumination, computing formula: relative variation=(numerical value of shading treatment-normal illumination numerical value) * 100%, with the leaf area of material to be identified, the relative variation of plant height and the contrast of low light tolerance are carried out relatively; Low temperature resistant authentication method is: select the strong and weak tomato of cold resistance to be contrast respectively, in the material to be identified sowing same period and plastics nutritive cube, management routinely, when tomato long to the 4th true leaf launch, 5 true leaves are just current, every part of material is got 20 strains and is moved on under 2 ℃ of dark or the low light condition, after 48 hours, rank is damaged to plants caused by sudden drop in temperature in investigation; Select the tomato that cold resistance is strong and cold resistance is weak to be contrast respectively, with grow seedlings in the material to be identified sowing same period and the plastics nutritive cube, field planting is in booth or greenhouse, manage routinely, remove first inflorescence, when first standard-sized sheet of using up treating second inflorescence put, get its pollen, on solid culture medium, observe down the sprouting situation in 8-12 ℃ of low temperature, medium making: with 80ml/L boric acid, 0.8% agar, after 10% sucrose medium was boiled mixing, adding one dripped on the spill slide and is made into solid culture medium, put into 8-12 ℃ growth case, after the equitemperature balance, the pollen that picks new collection with hair is broadcast in media surface, and it is capable to broadcast 2-4, examined under a microscope once every 15 minutes, measure pollen tube length and add up pollen germination rate and the length of measurement pollen tube with micrometer.
2, anti-multiple diseases of tomato according to claim 1 and breeding for stress tolerance authenticate technology method is characterized in that: the various disease state of an illness investigation grade scales that anti-multiple diseases is identified are:
(1) leaf mold state of an illness investigation grade scale, individual plant state of an illness grade scale:
0 grade: asymptomatic;
1 grade: the inoculation leaf chlorisis occurs to yellow scab;
3 grades: produce a small amount of white hypha, no spore on the inoculation leaf disease spot;
5 grades: produce mycelia and spore on the inoculation leaf disease spot;
7 grades: produce a large amount of mycelia and a large amount of spores on the inoculation leaf disease spot, and mycelia develops into upper leaf, but do not have spore;
9 grades: producing on the inoculation leaf disease spot has a large amount of spores, and spore also appears in upper leaf;
The disease-resistant grade scale of colony:
Immunity (I): do not infect, disease index is 0
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜15
Anti-disease (T): 15＜disease index＜30
Susceptible (S): disease index＞30
(2) the root knot nematode disease sentiment grade standard of checking the mark:
Individual plant state of an illness grade scale:
0 grade: unrooted tie lines worm and pieces of an egg
1 grade: 1-2 root knot and pieces of an egg
2 grades: 3-10 root knot and pieces of an egg
3 grades: 11-30 root knot and pieces of an egg
4 grades: 31-100 root knot and pieces of an egg
5 grades: 100 above root knots and pieces of an egg
The disease index computational methods
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is=0,
High anti-(HR): 0＜disease index＜1.0
Disease-resistant (R): 1.0＜disease index＜2.0
Anti-disease (T): 2.0＜disease index＜3.0
Susceptible (S): 3.0＜disease index＜4.0
High sense (HS): disease index＞4.0
(3) verticillium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: 1-2 sheet true leaf or two cotyledon flavescence, so that come off,
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging,
3 grades: complete stool is seriously wilted, so that withered,
The disease-resistant grade scale of colony
Immunity (I): do not infect, disease index is 0,
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
In anti-(MR): 30＜disease index＜40
Anti-disease (T): 40＜disease index＜60
Susceptible (S): disease index＞60
(4) fusarium wilt state of an illness investigation grade scale
Individual plant state of an illness grade scale:
0 grade: asymptomatic
1 grade: a slice true leaf or two cotyledon flavescence, so that come off,
2 grades: true leaf flavescence of 1-2 sheet or complete stool flavescence, leaf withering is sagging,
3 grades: complete stool is seriously wilted, so that withered,
The disease-resistant grade scale of colony
High anti-(HR): 0＜disease index＜10
Disease-resistant (R): 10＜disease index＜30
Susceptible (S): disease index＞30
The compound criteria for classification that infects of TMV, CMV
The compound individual plant criteria for classification that infects of TMV, CMV
0 grade: asymptomatic
1 grade: leaf is not open and flat, light floral leaf
3 grades: the serious shrinkage of blade, heavy floral leaf
5 grades: heavy floral leaf, blade slightly is out of shape, the fern leaf
7 grades: heavy floral leaf, gross distortion, line leaf
9 grades: plant stops growing or is close to dead
(5) TMV, the disease-resistant grade scale of CMV colony
High anti-(HR): 0＜disease index＜5
Disease-resistant (R): 5＜disease index＜10
Anti-disease (T): 10＜disease index＜30
Susceptible (S): disease index＞30
3, anti-multiple diseases of tomato according to claim 1 and 2 and breeding for stress tolerance authenticate technology method is characterized in that: the low light tolerance authentication method
Seedling stage, low light tolerance thought that regression equation is:
1. y=-0.7029x1-0.0550 x1: the thick ratio of plant height and stem relatively
2. y=-0.7173x2-0.0869 x2: chlorophyll b/a value relatively
3. y=-0.596x1-0.303x2-0.015 y: low light tolerance index
The phase low light tolerance regression equation of bearing fruit of blooming is:
4. y=-2.21x1+1.104 x1: the thick ratio of plant height and stem relatively
5. y=--0.524x2-0.1757 x2: chlorophyll b/a value relatively
Y: low light tolerance index
4, anti-multiple diseases of tomato according to claim 1 and 2 and breeding for stress tolerance authenticate technology method is characterized in that: in the low temperature resistant authentication method
(1) the low temperature identification of indicator of tomato seeds germination period is a seed germination index under 15 ℃ of conditions;
(2) do anti-cold direct indicator with the preceding two fringes relative fruit-setting rate really of seedling after the anti-cold exponential sum low temperature treatment, carry out correlation analysis simultaneously, think that relative conductivity, soluble sugar and soluble protein can be used as the indirect index of lower temperature resistance
Y=-0.6380x+2.6272 x: relative conductivity
Y=0.3397x+1.5130 x: relative protein content
(3) the bloom identification of indicator of the phase of bearing fruit is a pollen viability after low temperature treatment meiosis stage, seedling stage, bud phase lower temperature resistance unanimity, and the florescence is inconsistent,
The classification that tomato is damaged to plants caused by sudden drop in temperature
0 grade: plant leaf is normal, is damaged to plants caused by sudden drop in temperature;
1 grade: only the minority blade edge has slight shrinkage to wilt;
2 grades: the following blade of half is wilted dead;
3 grades: blade more than half is wilted dead;
4 grades: plant is all dead,
According to the rank that damages to plants caused by sudden drop in temperature of investigation, calculate each material and damage to plants caused by sudden drop in temperature the anti-cold finger number of exponential sum,
Damage to plants caused by sudden drop in temperature index=∑ xa/ (n ∑ x)=(X1a1+X2a2)/nT
X1, X2 in the formula, X3 ... Xn represents the tomato numbers that damage to plants caused by sudden drop in temperature at different levels, a1, a2, a3 ... an represents respectively to damage to plants caused by sudden drop in temperature grade, and T is total book of investigation.Anti-cold finger number is the inverse that damages to plants caused by sudden drop in temperature index.
5, anti-multiple diseases of tomato according to claim 3 and breeding for stress tolerance authenticate technology method is characterized in that: in the low temperature resistant authentication method
(1) the low temperature identification of indicator of tomato seeds germination period is a seed germination index under 15 ℃ of conditions;
(2) do anti-cold direct indicator with the preceding two fringes relative fruit-setting rate really of seedling after the anti-cold exponential sum low temperature treatment, carry out correlation analysis simultaneously, think that relative conductivity, soluble sugar and soluble protein can be used as the indirect index of lower temperature resistance
Y=-0.6380x+2.6272 x: relative conductivity
Y=0.3397x+1.5130 x: relative protein content
(3) the bloom identification of indicator of the phase of bearing fruit is: the pollen viability after the meiosis stage low temperature treatment, and seedling stage, bud phase lower temperature resistance unanimity, and the florescence is inconsistent,
The classification that tomato is damaged to plants caused by sudden drop in temperature
0 grade: plant leaf is normal, is damaged to plants caused by sudden drop in temperature;
1 grade: only the minority blade edge has slight shrinkage to wilt;
2 grades: the following blade of half is wilted dead;
3 grades: blade more than half is wilted dead;
4 grades: plant is all dead,
According to the rank that damages to plants caused by sudden drop in temperature of investigation, calculate each material and damage to plants caused by sudden drop in temperature the anti-cold finger number of exponential sum,
Damage to plants caused by sudden drop in temperature index=∑ xa/ (n ∑ x)=(X1a1+X2a2)/nT
X1, X2 in the formula, X3 ... Xn represents the tomato numbers that damage to plants caused by sudden drop in temperature at different levels, a1, a2, a3 ... an represents respectively to damage to plants caused by sudden drop in temperature grade, and T is total book of investigation, and anti-cold finger number is the inverse that damages to plants caused by sudden drop in temperature index.
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101408540B (en) * | 2008-11-17 | 2013-07-10 | 浙江省农业科学院 | Method for rapidly identifying resistance of watermelon and muskmelon varieties gummy stem blight |
| CN103270893A (en) * | 2013-05-31 | 2013-09-04 | 湖北省农业科学院经济作物研究所 | Method for appraising resistance of strawberries to gray mold |
| CN107980728A (en) * | 2017-11-29 | 2018-05-04 | 山东省花生研究所 | M hapla Resistance Identification method |
| CN108377893A (en) * | 2018-02-13 | 2018-08-10 | 东北农业大学 | A kind of rapid detection method of eggplant verticillium wilt disease resistance |
| CN108663481A (en) * | 2017-04-01 | 2018-10-16 | 浙江省农业科学院 | A kind of field symptom fast diagnosis method of tomato yellow leaf droop |
| CN109609589A (en) * | 2018-12-20 | 2019-04-12 | 东北农业大学 | A kind of method that Tomato gray leaf spot bacterium conidium stablizes sprouting and identifies for artificial inoculation on seedling |
| CN111886963A (en) * | 2020-07-30 | 2020-11-06 | 山东永盛农业发展有限公司 | Method for evaluating low-temperature-resistant variety of solanaceous vegetables |
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Cited By (8)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101408540B (en) * | 2008-11-17 | 2013-07-10 | 浙江省农业科学院 | Method for rapidly identifying resistance of watermelon and muskmelon varieties gummy stem blight |
| CN103270893A (en) * | 2013-05-31 | 2013-09-04 | 湖北省农业科学院经济作物研究所 | Method for appraising resistance of strawberries to gray mold |
| CN103270893B (en) * | 2013-05-31 | 2014-06-25 | 湖北省农业科学院经济作物研究所 | Method for appraising resistance of strawberries to gray mold |
| CN108663481A (en) * | 2017-04-01 | 2018-10-16 | 浙江省农业科学院 | A kind of field symptom fast diagnosis method of tomato yellow leaf droop |
| CN107980728A (en) * | 2017-11-29 | 2018-05-04 | 山东省花生研究所 | M hapla Resistance Identification method |
| CN108377893A (en) * | 2018-02-13 | 2018-08-10 | 东北农业大学 | A kind of rapid detection method of eggplant verticillium wilt disease resistance |
| CN109609589A (en) * | 2018-12-20 | 2019-04-12 | 东北农业大学 | A kind of method that Tomato gray leaf spot bacterium conidium stablizes sprouting and identifies for artificial inoculation on seedling |
| CN111886963A (en) * | 2020-07-30 | 2020-11-06 | 山东永盛农业发展有限公司 | Method for evaluating low-temperature-resistant variety of solanaceous vegetables |
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