CN100564543C - A kind of tomato parent pure line selection method of anti-Meloidogyne incognita - Google Patents
A kind of tomato parent pure line selection method of anti-Meloidogyne incognita Download PDFInfo
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- CN100564543C CN100564543C CNB2006101044271A CN200610104427A CN100564543C CN 100564543 C CN100564543 C CN 100564543C CN B2006101044271 A CNB2006101044271 A CN B2006101044271A CN 200610104427 A CN200610104427 A CN 200610104427A CN 100564543 C CN100564543 C CN 100564543C
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Abstract
The invention discloses a kind of selection of anti-Meloidogyne incognita tomato parent pure lines, but maternal C seeds of hybridized tomato A not anti-root knot nematode good with comprehensive proterties is the improvement object, seeds of hybridized tomato B with anti-root knot nematode is anti-source, utilize the anti-root knot nematode material of molecular marker screening of resistant gene in seedling stage, utilize traditional breeding method to select the comprehensive The Characters excellent material in field in the land for growing field crops, seed selection comprises that antagonism source B carries out molecular markers for identification and become strain phase sick garden resistance to identify in seedling stage.Is that maternal, B is that male parent is carried out artificial pollination with A, and with the maternal C of A as 5～6 generations of recurrent parent continuous backcross, 2 generations of selfing then.Each all utilizes the CAPs molecule marker antagonism root knot nematode proterties of the anti-root knot nematode Mi of tomato gene to screen in seedling stage from generation to generation, carries out disease garden assistant identification and economical character in the field and selects.The tomato parent pure lines that contain the resistant gene that isozygotys, have different outstanding good economical characters have been selected at last.
Description
Technical field
The invention belongs to the tomato breeding field, the selection that relates to a kind of tomato, be particularly related to a kind of tomato parent pure line selection method of anti-Meloidogyne incognita, this method selects breeding method to combine with molecular marker assisted selection and tradition, and the parent's pure lines that filter out can be directly used in the combo of tomato of preventing root-knot Nematode disease cross-fertilize seed.
Background technology
Root knot nematode (Meloidogyne spp.) is one of very serious soil-borne disease of harm cultivation tomato (Lycoperisicon esculentum), and its main kind is Meloidogyne incognita (Meloidogyneincognita).Along with the expansion of tomato protection ground cultivated area and the raising of multiple crop index, the harm of root knot nematode is serious day by day, and the production loss in morbidity field is generally 30%～50%, reaches more than 70% when serious.For chemical prevention and the biological control of the root knot nematode of tomato, because pollution of ecological environment or because soil fungistasis, its ecological benefits and prevention effect are all undesirable.Therefore, seed selection and utilize the resistance tomato variety just to become the valid approach of control root knot nematode, but the domestic tomato variety of non-resistant still is used for producing at present.Resistant variety breed excavation and the utilization that depends on the resistance resource, traditional artificial inoculation authentication method not only program complexity, result is inaccurate, and very difficult homozygote and the heterozygote of distinguishing resistant gene, cause the speed of isozygotying of resistant gene slower, seriously restricting the seed selection of tomato anti-root knot nematode parent system and new variety.
Summary of the invention
At the problem that exists in above-mentioned tomato of preventing root-knot Nematode disease parent system and the breed breeding, the objective of the invention is to, a kind of tomato parent pure line selection method of anti-Meloidogyne incognita is proposed, this method utilizes molecular marker assisted selection and tradition to select the breeding seed selection tomato of preventing root-knot Nematode disease parent system that combines, easy and simple to handle, reliable results, the parent who is selected pure lines can be directly used in the combo of tomato of preventing root-knot Nematode disease cross-fertilize seed.
To achieve these goals, the present invention adopts following technical solution:
A kind of selection of anti-Meloidogyne incognita tomato parent pure lines, it is characterized in that, this method is good with comprehensive proterties but maternal C seeds of hybridized tomato A not anti-root knot nematode is the improvement object, seeds of hybridized tomato B with anti-root knot nematode is anti-source, utilize the anti-root knot nematode material of molecular marker screening of resistant gene in seedling stage, utilize traditional breeding method to select the comprehensive The Characters excellent material in field in the land for growing field crops, concrete seed selection comprises the following steps:
1) molecular markers for identification and become strain phase sick garden resistance to identify in seedling stage is carried out in the antagonism source
On the basis of molecular markers for identification, will resist the source to transplant in the root knot nematode disease garden, the antagonism source is carried out to strain phase sick garden resistance and identifies;
Select molecular markers for identification and becomes the disease garden resistance evaluation of strain phase to determine to carry Mi gene and disease-resistant anti-source in seedling stage as transformation;
2) good but seeds of hybridized tomato A not anti-root knot nematode is a female parent with comprehensive proterties, be that male parent is carried out artificial pollination with the seeds of hybridized tomato B of anti-root knot nematode, results cross-fertilize seed ABF
1, carry out individual plant DNA extraction and pcr amplification, keep the plant that amplifies the 750bp band, eliminate the individual plant of no amplified band;
Pcr amplification product to selected individual plant DNA is cleared up with nucleic acid restriction endonuclease Taq I, and the SCAR mark is converted into the CAPs mark; After endonuclease reaction and electrophoresis detection, eliminate the individual plant that does not contain the Mi gene, keep Mi gene pure strain and the strain of Mi genetic heterozygosis;
3) will be selected in plant and be colonizated in root knot nematode disease garden, field, and carry out the evaluation that disease garden assistant identification and plant type, first inflorescence Main Agronomic Characters such as tight knot position, fruit look, fruit shape, resistance and select, the plant with outstanding good character will be reserved seed for planting by individual plant;
4) good with comprehensive proterties but maternal C seeds of hybridized tomato A not anti-root knot nematode is a recurrent parent, with selected individual plant is nonrecurrent parent, 5～6 generations of continuous backcross, per generation continue to carry out as stated above seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select, selected anti-root knot nematode and have the individual plant or the strain system of outstanding good character, select 3～5 strains to be according to the different characteristics of good character is final, by strain mooring kind;
5) it is 2 generations of continuous selfing to going into roguing, proceed seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select the final tomato parents system that obtains to contain the resistant gene that isozygotys, has different outstanding good economical characters.
The present invention brings following technique effect:
(1) the present invention is applied to molecular marking technique the screening of tomato breeding material first, and combines with traditional breeding way, has solved loaded down with trivial details, the inaccurate problem of qualification result of tomato root-knot nematode resistant garden qualification program effectively.
(2) be base mateiral with the seeds of hybridized tomato that utilizes in 2 productions, help the accumulation of beneficial gene, guarantee that the parent plant cording that is obtained has high combining ability.
(3) the maternal C with the A kind backcrosses many generations, at its self progeny's that backcrosses the good character of having strengthened C in the roguing system of going into.
Description of drawings
Fig. 1 is the expanding effect figure of the anti-root knot nematode SCAR of tomato mark.The the 1st and the 8th swimming lane is Marker; The the 2nd～the 3rd swimming lane does not have the 750bp band, is considered as not containing the Mi resistant gene; The 750bp band has all appearred in the 4th～the 7th swimming lane, need further carry out the CAPs labeled analysis.
Fig. 2 is the restriction enzyme digestion and electrophoresis figure of the anti-root knot nematode CAPs of tomato mark.The M swimming lane is Marker; The the 1st～the 4th swimming lane enzyme is cut product and 750bp, 570bp and 160bp three bands are occurred, expression Mi genetic heterozygosis; The 5th swimming lane enzyme is cut product and 570bp and 160bp two bands are occurred, expression Mi gene pure.The the 6th～the 8th swimming lane has only 750bp one band, and expression does not contain the Mi gene.
Fig. 3 is that M02-28-3-6-12-69-158 and contrast (left side) identify that at resistant to southern root knot nematode the incidence in the garden contrasts for the parent.
Fig. 4 identifies that at resistant to southern root knot nematode the incidence in the garden contrasts with contrast (left side) for M02-69-9-62-32-49-216 parent system.
Fig. 5 identifies that at resistant to southern root knot nematode the incidence in the garden contrasts with contrast (left side) for M02-58-9-4-82-89-213 parent system.
The present invention is described in further detail below in conjunction with embodiment that accompanying drawing and contriver provide.
Embodiment
The present invention is applied to molecular marking technique the screening of tomato breeding material first, utilize the anti-root knot nematode material of molecular marker screening of resistant gene in seedling stage, utilize traditional breeding method to select the comprehensive The Characters excellent material in field in the land for growing field crops, finally select anti-root knot nematode and have the parent pure lines of the tomato strain system of outstanding good character as cross-fertilize seed of new generation.Its concrete steps are as follows:
(1) produces at present with China that to go up the comprehensive proterties of using good but maternal C (seeds of hybridized tomato A and maternal C thereof can ask for from the unit or its production of hybrid seeds unit that breed seeds of hybridized tomato) seeds of hybridized tomato A not anti-root knot nematode mainly improves object, to originate for resistance from the seeds of hybridized tomato B of the anti-root knot nematode of external introduction (seed is sold retail sales or the company of seed import and export business is arranged from the market, buys as units such as Chinese Seed Group Co., Shanghai kind industry (group) company limited and Wal, Shanghai agricultural science and technology company limiteds).
(2) antagonism source B carries out molecular markers for identification and become strain phase sick garden resistance to identify in seedling stage.Concrete grammar is: sow anti-source B earlier, when treating that its seedling grows to 2～3 true leaves, and total DNA of employing micromethod extraction single-strain blade (Gong Zhenhui etc. are with the minim DNA extracting method of PCR evaluation transfer-gen plant. Northwest Agricultural University's journal, 1997,25 (1): 45～48).SCAR flag sequence according to the anti-root knot nematode Mi of the tomato of having published gene designs forward and reverse primer (Williamson V M, Ho J Y, WuF F, et al.A PCR-based marker tightly linked to the nematode resistance gene, Mi, in tomato.Theoretical and Applied Genetics, 1994,87:757～763), the forward primer sequence is: 5 '-TCGGAGCCTTGG TCTGAATT3 ', the reverse primer sequence is: 5 '-GCCAGAGATG ATTCGTGAGA-3 '.PCR reaction cumulative volume is 15 μ L, comprising: template DNA 2 μ L, forward primer (20 μ M) 0.5 μ L, reverse primer (20 μ M) 0.5 μ L, dNTP (2mM) 1.5 μ L, (20mM contains Mg to 10 * buffer
2+) 1.5 μ L, ddH
2O 8.5 μ L, Taq enzyme (2U/ μ L) 0.5 μ L.The PCR response procedures is: 95 ℃, and 5min; 94 ℃, 1min, 59 ℃, 1min, 72 ℃, 2min, 30 circulations; 72 ℃, 7min.Utilize total DNA of this primer antagonism source individual plant to carry out pcr amplification, plant can amplify the band of a treaty 750bp; Clear up pcr amplification product with nucleic acid restriction endonuclease TaqI again, the SCAR mark is converted into the CAPs mark.It is 10 μ L that enzyme is cut the system cumulative volume, comprising amplified production 7 μ L, and 5U/ μ L Taq I restriction enzyme 0.5 μ L, 10 * buffer1.0 μ L, ddH
2O 1.5 μ L.Reaction mixture is at 65 ℃ of incubation 1h of PCR instrument.After the electrophoresis detection, do not had only 750bp one band by what enzyme was cleared up, these individual plants do not contain the Mi gene; Can two or three enzyme bands be occurred by what enzyme was cleared up, what 570bp and two specific fragments of 160bp wherein occur is the strain of Mi gene pure, and 750bp, 570bp and three specific fragments of 160bp appear be the strain of Mi genetic heterozygosis.Strain of Mi gene pure or heterozygosis strain all can be used as anti-source.
On the basis of molecular markers for identification, will resist the source transplanting to be carried out to strain phase sick garden resistance in antagonism source, root knot nematode disease garden and identify.
Have only seedling stage molecular markers for identification with become strain phase sick garden resistance to identify to determine to carry Mi gene and the disease-resistant anti-source that can be used as transformation.
(3) be that female parent, B are that male parent is carried out artificial pollination with A, results cross-fertilize seed ABF
1Carry out individual plant DNA extraction and pcr amplification as stated above.Overwhelming majority plant has amplified the band of a treaty 750bp; The small part plant does not have amplified production, is considered as not containing root knot nematode Mi resistant gene (accompanying drawing 1), eliminates the individual plant of no amplified band.
The pcr amplification product of selected individual plant DNA is cleared up with nucleic acid restriction endonuclease Taq I, and the SCAR mark is converted into the CAPs mark.Endonuclease reaction is undertaken by (2) described enzyme blanking method in this specification sheets " embodiment ".After the electrophoresis detection, do not had only 750bp one band by what enzyme was cleared up, these individual plants do not contain the Mi gene, are eliminated; Can two or three enzyme bands be occurred by what enzyme was cleared up, what 570bp and two specific fragments of 160bp wherein occur is the strain of Mi gene pure, and 750bp, 570bp and three specific fragments of 160bp appear be Mi genetic heterozygosis strain (accompanying drawing 2).The selected resistant gene individual plant with heterozygosis that isozygotys.
To be selected in plant and be colonizated in root knot nematode disease garden, field, and carry out the evaluation that disease garden assistant identification and plant type, first inflorescence Main Agronomic Characters such as tight knot position, fruit look, fruit shape, resistance and select, the plant with outstanding good character will be reserved seed for planting by individual plant.
(4) the maternal C with the A kind is a recurrent parent, with selected individual plant is nonrecurrent parent, 5～6 generations of continuous backcross, per generation continue to carry out as stated above seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select selected anti-root knot nematode and have the individual plant or the strain system of outstanding good character.Finally can select 3～5 strain systems according to the different characteristics of good character, by strain mooring kind.
(5) it is 2 generations of continuous selfing to going into roguing, proceed seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select the final tomato parents system that obtains to contain the resistant gene that isozygotys, has different outstanding good economical characters.
Below be the specific embodiment that the contriver provides, but the present invention is not limited to this embodiment.
Embodiment:
(1) good but seeds of hybridized tomato not anti-Meloidogyne incognita " No. one, golden canopy " is a female parent (A kind with the comprehensive proterties of China, provide by seedling company limited of Xi'an Jinpeng), hybridize for male parent with cross-fertilize seed Kelly (B kind, just reaching earlier kind of an industry company limited by Shouguang, Shandong provides) from containing of external introduction of anti-tomato root-knot eelworm gene M i.
(2) antagonism source B carries out molecular markers for identification and become strain phase sick garden resistance to identify in seedling stage.Concrete grammar is: sow anti-source B earlier, when treating that its seedling grows to 2～3 true leaves, and total DNA of employing micromethod extraction single-strain blade (Gong Zhenhui etc. are with the minim DNA extracting method of PCR evaluation transfer-gen plant. Northwest Agricultural University's journal, 1997,25 (1): 45～48).SCAR flag sequence according to the anti-root knot nematode Mi of the tomato of having published gene designs forward and reverse primer (Williamson V M, Ho J Y, WuF F, et al.A PCR-based marker tightly linked to the nematode resistance gene, Mi, in tomato.Theoretical and Applied Genetics, 1994,87:757～763), the forward primer sequence: 5 '-TCGGAGCCTTGG TCTGAATT-3 ', the reverse primer sequence: 5 '-GCCAGAGATG ATTCGTGAGA-3 '.PCR reaction cumulative volume is 15 μ L, comprising: template DNA 2 μ L, forward primer (20 μ M) 0.5 μ L, reverse primer (20 μ M) 0.5 μ L, dNTP (2mM) 1.5 μ L, (20mM contains Mg to 10 * buffer
2+) 1.5 μ L, ddH
2O 8.5 μ L, Taq enzyme (2U/ μ L) 0.5 μ L.The PCR response procedures is: 95 ℃, and 5min; 94 ℃, 1min, 59 ℃, 1min, 72 ℃, 2min, 30 circulations; 72 ℃, 7min.Utilize total DNA of this primer antagonism source individual plant to carry out pcr amplification, plant can amplify the band of a treaty 750bp; Clear up pcr amplification product with nucleic acid restriction endonuclease TaqI again, the SCAR mark is converted into the CAPs mark.It is 10 μ L that enzyme is cut the system cumulative volume, comprising amplified production 7 μ L, and 5U/ μ L Taq I restriction enzyme 0.5 μ L, 10 * buffer1.0 μ L, ddH
2O 1.5 μ L.Reaction mixture is at 65 ℃ of incubation 1h of PCR instrument.After the electrophoresis detection, do not had only 750bp one band by what enzyme was cleared up, these individual plants do not contain the Mi gene; Can two or three enzyme bands be occurred by what enzyme was cleared up, what 570bp and two specific fragments of 160bp wherein occur is the strain of Mi gene pure, and 750bp, 570bp and three specific fragments of 160bp appear be the strain of Mi genetic heterozygosis.Strain of Mi gene pure or heterozygosis strain all can be used as anti-source.
On the basis of molecular markers for identification, will resist the source transplanting to be carried out to strain phase sick garden resistance in antagonism source, root knot nematode disease garden and identify.
By seedling stage molecular markers for identification with become strain phase sick garden resistance to identify to determine that Kelly carries Mi gene and disease-resistant, can be used as the anti-source of transformation.
(3) be female parent with " No. one, golden canopy " A kind, " Kelly " B kind is that male parent is carried out artificial pollination, results cross-fertilize seed ABF
1Carry out individual plant DNA extraction and pcr amplification as stated above.Overwhelming majority plant has amplified the band of a treaty 750bp; The small part plant does not have amplified production, is considered as not containing root knot nematode Mi resistant gene (accompanying drawing 1).Eliminate the individual plant of no amplified band.
The pcr amplification product of selected individual plant DNA is cleared up with nucleic acid restriction endonuclease Taq I, and the SCAR mark is converted into the CAPs mark.Endonuclease reaction carries out as stated above.After the electrophoresis detection, do not had only 750bp one band by what enzyme was cleared up, these individual plants do not contain the Mi gene, are eliminated; Can two or three enzyme bands be occurred by what enzyme was cleared up, what 570bp and two specific fragments of 160bp wherein occur is the strain of Mi gene pure, and 750bp, 570bp and three specific fragments of 160bp appear be the strain of Mi genetic heterozygosis.The selected resistant gene individual plant with heterozygosis that isozygotys.
To be selected in plant and be colonizated in root knot nematode disease garden, field, and carry out the evaluation that disease garden assistant identification and plant type, first inflorescence Main Agronomic Characters such as tight knot position, fruit look, fruit shape, resistance and select, the plant with outstanding good character will be reserved seed for planting by individual plant.
(4) with maternal T15-28-6 (the maternal C of A kind of " No. one, golden canopy ", provide by seedling company limited of Xi'an Jinpeng) be recurrent parent, with selected individual plant is nonrecurrent parent, 5～6 generations of continuous backcross, per generation continue to carry out as stated above seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select selected anti-root knot nematode and have the individual plant or the strain system of outstanding good character.Different characteristics according to good character finally has been selected in 3 strain systems, by strain mooring kind.
(5) it is 2 generations of continuous selfing to going into roguing, proceed seedling stage the genjie eelworm resistance gene molecular marker screening and the field economical character select, bred three good parents systems at last, named and be M02-28-3-6-12-69-158, M02-69-9-62-32-49-216 and M02-58-9-4-82-89-213.
With above-mentioned 3 good parents is to be colonizated in the tomato root-knot eelworm resistance to identify the garden, and the result shows this three parents system all high anti-Meloidogyne incognitas (accompanying drawing 3, Fig. 4, Fig. 5).The main characteristic of three parent systems that breed is as follows:
M02-28-3-6-12-69-158: the pink fruit type of limited growth.The plant strain growth gesture is stronger, plant height 60～65cm, and blade is rarer.Stem the 6th leaf and is given birth to first inflorescence, and three to four layers bind.Fruit is high circular, and young fruit does not have green shoulder, and glossiness is good, mellow fruit pink, general single fruit weight 200～250g.The hardness of fruit is good, and shelf-life is long, storage tolerance.High anti-root knot nematode and Tomato mosaic virus.
M02-69-9-62-32-49-216: the pink fruit type of indeterminate growth.The plant strain growth gesture is stronger, and blade is rarer.Stem the 7th leaf and is given birth to first inflorescence, every three leaves later on and gives birth to an inflorescence.Fruit is high circular, and young fruit does not have green shoulder, and smooth surface is shinny, and the mellow fruit pink is about general single fruit weight 250g.Pulp is thick, and ventricle is big, and hardness is good, and shelf-life is long, storage tolerance.High anti-root knot nematode and blight.
M02-58-9-4-82-89-213: the bright red fruit type of indeterminate growth.The plant strain growth gesture is stronger, and blade is rarer, and internode is longer.Stem the 7th leaf and is given birth to first inflorescence, every three leaves later on and gives birth to an inflorescence.Fruit is high circular, and young fruit does not have green shoulder, the shinny nothing wrinkle of fruit surface smooth, and the mellow fruit pink is about general single fruit weight 250g.Pulp is thick, and hardness is big, shelf-life 10～15 days.High anti-root knot nematode, leaf mold and Tomato mosaic virus.
Claims (1)
1. the selection of anti-Meloidogyne incognita tomato parent pure lines, it is characterized in that, this method is good with comprehensive proterties but maternal C seeds of hybridized tomato A not anti-root knot nematode is the improvement object, seeds of hybridized tomato B with anti-root knot nematode is the resistance source, utilize the anti-root knot nematode material of molecular marker screening of resistant gene in seedling stage, utilize traditional breeding method to select the comprehensive The Characters excellent material in field in the land for growing field crops, concrete seed selection comprises the following steps:
1) molecular markers for identification and become strain phase sick garden resistance to identify in seedling stage is carried out in the antagonism source
On the basis of molecular markers for identification, will resist the source to transplant in the root knot nematode disease garden, the antagonism source is carried out to strain phase sick garden resistance and identifies;
Selection by seedling stage molecular markers for identification determine to carry the Mi gene and become strain phase sick garden resistance to identify the anti-source of determining to have disease-resistant proterties as transformation;
2) good but seeds of hybridized tomato A not anti-root knot nematode is a female parent with comprehensive proterties, be that male parent is carried out artificial pollination with the seeds of hybridized tomato B of anti-root knot nematode, results cross-fertilize seed ABF
1, carry out individual plant DNA extraction and pcr amplification; The used forward and reverse primer of pcr amplification is according to the SCAR flag sequence design of the anti-root knot nematode Mi of the tomato gene of having published, the forward primer sequence: 5 '-TCGGAGCCTTGG TCTGAATT-3 ', the reverse primer sequence: 5 '-GCCAGAGATGATTCGTGAGA-3 '; Reservation can amplify the plant of 760bp band, eliminates the individual plant of no amplified band;
Pcr amplification product to selected individual plant DNA is cleared up with nucleic acid restriction endonuclease Taq I, and the SCAR mark is converted into the CAPs mark; After endonuclease reaction and electrophoresis detection, do not had only 750bp one band by what enzyme was cleared up, these individual plants do not contain the Mi gene, eliminate the individual plant that does not contain the Mi gene; Two or the three enzyme bands of appearance that can be cleared up by enzyme, what 570bp and two specific fragments of 160bp wherein occur is the strain of Mi gene pure, and occur 750bp, 570bp and three specific fragments of 160bp for the strain of Mi genetic heterozygosis, keep Mi gene pure strain and the strain of Mi genetic heterozygosis;
3) will be selected in plant and be colonizated in root knot nematode disease garden, field, and carry out the evaluation that disease garden assistant identification and plant type, first inflorescence tight knot position, fruit look, fruit shape, resistance economical character and select, the plant with outstanding good character will be reserved seed for planting by individual plant;
4) good with comprehensive proterties but maternal C seeds of hybridized tomato A not anti-root knot nematode is a recurrent parent, with selected individual plant is nonrecurrent parent, 5～6 generations of continuous backcross, per generation continue to carry out as stated above seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select, selected anti-root knot nematode and have the individual plant or the strain system of outstanding good character, select 3～5 strains to be according to the different characteristics of good character is final, by strain mooring kind;
5) it is 2 generations of continuous selfing to going into roguing, proceed seedling stage the genjie eelworm resistance gene molecular marker screening and the sick garden assistant identification in field and economical character select the final tomato parents system that obtains to contain the resistant gene that isozygotys, has different outstanding good economical characters.
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| CN102696476A (en) * | 2012-06-28 | 2012-10-03 | 抚顺市北方农业科学研究所 | Method for sorting homozygous pink-fruit-color and red-fruit-color tomato inbred lines by late-mature-gene tomato |
| MX360582B (en) | 2012-12-13 | 2018-11-07 | Inst De Ecologia A C Star | Biocontrol of phyto-parasitic nematodes using paecilomyces. |
| CN103999767A (en) * | 2014-06-05 | 2014-08-27 | 青岛农业大学 | Breeding method of tomato breeding material with resistance to root knot nematode disease and yellow leaf curl virus disease |
| CN104541875A (en) * | 2014-12-31 | 2015-04-29 | 安徽徽大农业有限公司 | Yellow cherry tomato scion cutting breeding method |
| CN107258341A (en) * | 2017-07-26 | 2017-10-20 | 桂平市蒙圩镇火炎种养专业合作社 | A kind of prevention and controls of canopy room tomato root-knot eelworm disease |
| CN111149690A (en) * | 2020-03-20 | 2020-05-15 | 山东永盛农业发展有限公司 | Method for cultivating tomato root-knot nematode-resistant salt-tolerant inbred line |
| CN113913438B (en) * | 2021-10-12 | 2023-11-14 | 中国农业科学院茶叶研究所 | Application of plant BIAF gene |
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| A PCR-based marker tightly linked to the nematode resistancegene,Mi,in tomato.. Williamson V M.Theoretical and Applied Genetics,Vol.87 . 1994 |
| A PCR-based marker tightly linked to the nematode resistancegene,Mi,in tomato.. Williamson V M.Theoretical and Applied Genetics,Vol.87 . 1994 * |
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| 利用PCR技术同时鉴定番茄抗根结线虫和抗斑萎病毒原因. 李君明等.园艺学报，第30卷第6期. 2003 * |
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