CN105993911A - Hybridization breeding method for high-yield maize - Google Patents
Hybridization breeding method for high-yield maize Download PDFInfo
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- CN105993911A CN105993911A CN201610303820.7A CN201610303820A CN105993911A CN 105993911 A CN105993911 A CN 105993911A CN 201610303820 A CN201610303820 A CN 201610303820A CN 105993911 A CN105993911 A CN 105993911A
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
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- A—HUMAN NECESSITIES
- A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
- A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
- A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
- A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
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- Genetics & Genomics (AREA)
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- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention discloses a hybridization breeding method for high-yield maize. The method comprises the following steps that good maize varieties are selected as maize male parents and maize female parents to be hybridized, and hybridized combination F1 generation seeds are obtained; under the screening planting condition, the obtained F1 generation seeds are selfed to form a second cycle line group, special plants with screening characteristics are selected from the second cycle line group to be selfed for 10 generations or above, and second female parents with the stable characteristics are obtained; DNA plasmids containing BT insecticidal toxin protein genes, bar genes, maize dwarf mosaic virus coat protein genes DMVCPDNA are introduced into seedling cells or tissue or organs of the second female parents through particle bombardment to be cultured and screened. According to the method, the second female parents of the maize are screened under the low-phosphorus, drought and saline-alkaline conditions, and the seeds which are high in phosphorus absorption effect and have the excellent insect resistance, grass resistance, disease resistance, drought resistance and saline-alkaline resistance are obtained through the synergistic effect of particle bombardment.
Description
Technical field
The present invention relates to corn breeding field, be specifically related to the cross breeding method of a kind of high-yield corn.
Background technology
Semen Maydis is very big at the sown area of China, and distribution is the widest, be north of China and Southwest Mountainous Areas and other
One of main food of the winterbourne area people.The good harvest of Semen Maydis is the weight being related to grain security and social stability
Big problem.In recent years, due to weather conditions such as arids, moisture can not be watered to Semen Maydis in time again, cause jade
The rice significantly underproduction.Simultaneously as phosphate fertilizer upward price trend in recent years, a lot of peasants in corn planting Shaoshi or
Not executing phosphate fertilizer, the drought tolerance that also result in Semen Maydis reduces and the underproduction.Therefore, high yield phosphorus efficiency, drought-enduring is cultivated
And the corn variety of Salt And Alkali Tolerance be the following important channel solving China's grain security and Environmental and resource issue it
One.
Corn material lacks effective natural resistance gene simultaneously, is difficult with traditional breeding technique means
Screening obtains the corn material with excellent insect resistace, and traditional Pest control measure is mainly chemical prevention,
But chemical pesticide has cost height, environmental pollution, easily residual, can poison beneficial insect and pest natural enemy etc.
Drawback is not suitable for life-time service.
The rise of plant genetic engineering and development, the preventing and treating for Corn diseases and insect pests provides a brand-new thinking.
Utilize genetic engineering means cultivate pest-resistant new varieties have free from environmental pollution, insect pesticidal is had single-minded
Property, breeding cycle are short, breeding and cultivation low cost and other advantages.What application genetic engineering means was cultivated now is anti-
Worm material quantity ramps, and almost covers all of important crops.In numerous anti insect genes,
Bt insecticidal protein gene is that a class Study on mechanism is clearer, and market is most widely used general and the most potential
Insecticidal proteins.Since 1993 in the world first case successfully turn cry1Ab gene pest-resistant Semen Maydis and occur, anti-
Worm transgenic corns quickly grows, until 1996, express the Transgenic Bt Corn quilt of cry1Ab gene
Approval business is applied to control European corn borer harm.Subsequently, carry and come from killing of bacillus thuringiensis
The transgenic corns of worm albumen is being increasingly used to control lepidoptera pest main in Maize Production.Tool
The corn germplasm having Cry1Ab protein expression has proven to one for the hybridization carrying out Semen Maydis and can subtract
Few Pesticide use, and reduce the effective control measures to environment and mankind's potential hazard.
Summary of the invention
For solving the problems referred to above, the invention provides the cross breeding method of a kind of high-yield corn, by low
Phosphorus, arid and saline and alkaline under the conditions of to screen Semen Maydis second maternal, and by particle bombardment by BT toxic protein
Gene, bar gene, Coat Protein Gene of Maize Dwarf Mosaic Virus MDMVCPDNA import the jade of the second female parent
In rice callus, thus it is high to obtain phosphorus assimilation effect, and has excellent insect resistace, anti-grass, resists
Characteristic of disease, drought tolerance, the seed of Salt And Alkali Tolerance.
For achieving the above object, the technical scheme that the present invention takes is:
The cross breeding method of a kind of high-yield corn, comprises the steps:
S1, select suitable local plantation, Resistant, the high-quality that anti-fall, tiller is weak, fringe is big, setting percentage is high,
High-yield variety hybridizes as Semen Maydis male parent and Semen Maydis female parent, obtains cross combination F1 generation seed；
S2, under the conditions of selection growth, utilize gained F1 generation seed selfing formed second cycle line colony, and
Therefrom select that there is special strain selfing 10 generation of screening character more than, obtain the second female parent that character is stable；Tool
Body, under the conditions of low-phosphorous, sow seed, between milpa length is to seven leaf phases to emulsified phase, to Semen Maydis
Plant carries out Osmotic treatment, no longer waters in corn-growing regions and pours water；And after milk stage in 1～2 day
In corn-growing regions, pouring saline-alkali water is to saline-alkali water above ground level 1～5cm, normally waters Semen Maydis afterwards and pours water
Point；
S3, will be containing BT insecticidal neurotoxin gene, bar gene, maize dwarf mosaic virus by particle bombardment
The DNA plasmid of coat protein gene MDMVCPDNA imports Seedling cell, tissue or the organ of above-mentioned second female parent,
After 75% ethanol surface sterilization, being placed on division culture medium, under the conditions of 25 DEG C～27 DEG C, lucifuge recovers training
Support one day, after cultivating 3 days under illumination, be transferred in 1/2MS+IBA 4.8mg/L+NAA 0.04mg/L culture medium
Continue to cultivate shading renewal cultivation one day；
S4, grow to more than 1cm to resistant buds after, move it into root media: improvement MS+0.04mg/L
Continuing in NAA+0.75mg/L 6-BA+0.075mg/L IBA+35g/L sucrose medium to cultivate, growth of seedling is extremely
After well developed root system, stem and leaf stalwartness, quality trait is measured comprehensively, screening yield is high, comprehensive resistance is strong,
Colory strain moves in greenhouse, and conventional cultivation, until producing seeds.
Wherein, every liter of saline-alkali water comprise 23.176 grams of sodium chloride, 3.16 grams of magnesium chloride, 2.148 grams of magnesium sulfate,
2.243 grams of calcium chloride, sodium bicarbonate 0.379 gram, 0.623 gram of potassium chloride.
Wherein, the improvement MS in institute's step S4 include manganese methionine 15mg/L, zinc-amino acid chelate 8mg/L,
Nucleoside hydrochlorate 13mg/L, magnesium amino acid chelate 6mg/L, Selenomethionine 5.5mg/L, phytase 0.85mg/L,
Pyridine acid chromium salt 0.065mg/L, calcium iodate 0.3mg/L, lysine 6mg/L, humic acid chelating cobalt
0.065mg/L, albumen germanium 0.75mg/L, iodine yeast 6mg/L, thiamine hydrochloride 10mg/L, hydrochloric acid pyrrole
Tremble alcohol 3mg/L, EM original solution 35mg/L, cyclohexanhexanol 27mg/L.
Wherein, 5 '-adenylic acid dipotassium that described nucleoside hydrochlorate obtains after including ribonucleic acid basic hydrolysis or sodium,
5 '-bird adenylic acid dipotassium or sodium, 5 '-cytidylic acid dipotassium or sodium and 5 '-uridylic acid dipotassium.
Wherein, described screening character includes any three kinds in following character:
Corn kernel: long awns, white shell and yellow；
Plant height: 100～110cm；
Fruit ear: tassel row number 10～18 row mass of 1000 kernel 200～300g, fecundity is good；
Fringe position is on the low side: 40～60cm.
Wherein, described step S2 prior to seeding, need to be at corn-growing regions applying base fertilizer: nitrogenous fertilizer 110kg N/hm2、
Phosphate fertilizer 5kg P2O5/hm2With potash fertilizer 12kg K2O/hm2, to form low-phosphorous condition.
The method have the advantages that
Maternal by screening Semen Maydis second under the conditions of low-phosphorous, arid and saline and alkaline, and will by particle bombardment
BT insecticidal neurotoxin gene, bar gene, Coat Protein Gene of Maize Dwarf Mosaic Virus MDMVCPDNA import
In the maize calli of the second female parent, thus it is high to obtain phosphorus assimilation effect, and have excellent insect resistace,
Anti-grass, disease resistance, drought tolerance, the seed of Salt And Alkali Tolerance.
Detailed description of the invention
In order to make objects and advantages of the present invention clearer, below in conjunction with embodiment, the present invention is carried out
Further describe.Should be appreciated that specific embodiment described herein only in order to explain the present invention,
It is not intended to limit the present invention.
In following example, the character standard of screening includes any three kinds in following character: corn kernel: long
Awns, white shell and yellow；Plant height: 100～110cm；Fruit ear: tassel row number 10～18 row mass of 1000 kernel 200～
300g, fecundity is good；Fringe position is on the low side: 40～60cm.The nucleoside hydrochlorate used includes ribonucleic acid aqueous alkali
5 ' obtained after solution-adenylic acid dipotassium or sodium, 5 '-bird adenylic acid dipotassium or sodium, 5 '-cytidylic acid dipotassium or
Sodium and 5 '-uridylic acid dipotassium；The every liter of saline-alkali water used comprises 23.176 grams of sodium chloride, magnesium chloride 3.16
Gram, 2.148 grams of magnesium sulfate, 2.243 grams of calcium chloride, sodium bicarbonate 0.379 gram, 0.623 gram of potassium chloride.Institute
Use improvement MS include manganese methionine 15mg/L, zinc-amino acid chelate 8mg/L, nucleoside hydrochlorate 13mg/L,
Magnesium amino acid chelate 6mg/L, Selenomethionine 5.5mg/L, phytase 0.85mg/L, pyridine acid chromium salt 0.065mg/L,
Calcium iodate 0.3mg/L, lysine 6mg/L, humic acid chelating cobalt 0.065mg/L, albumen germanium 0.75mg/L,
Iodine yeast 6mg/L, thiamine hydrochloride 10mg/L, pyridoxine hydrochloride 3mg/L, EM original solution 35mg/L, ring
Hexanhexol 27mg/L.
Embodiment
S1, select suitable local plantation, Resistant, the high-quality that anti-fall, tiller is weak, fringe is big, setting percentage is high,
High-yield variety hybridizes as Semen Maydis male parent and Semen Maydis female parent, obtains cross combination F1 generation seed；
S2, under the conditions of selection growth, utilize gained F1 generation seed selfing formed second cycle line colony, and
Therefrom select that there is special strain selfing 10 generation of screening character more than, obtain the second female parent that character is stable；Tool
Body, prior to seeding, need to be at corn-growing regions applying base fertilizer: nitrogenous fertilizer 110kg N/hm2, phosphate fertilizer 5kg P2O5/hm2
With potash fertilizer 12kg K2O/hm2, to form low-phosphorous condition；Seed is sowed, milpa under the conditions of low-phosphorous
Between length is to seven leaf phases to emulsified phase, milpa is carried out Osmotic treatment, no longer waters in corn-growing regions
Pour water；And after milk stage in 1～2 day in corn-growing regions water saline-alkali water exceed ground to saline-alkali water
Face 1～5cm, afterwards pouring moisture normal to Semen Maydis；
S3, will be containing BT insecticidal neurotoxin gene, bar gene, maize dwarf mosaic virus by particle bombardment
The DNA plasmid of coat protein gene MDMVCPDNA imports Seedling cell, tissue or the organ of above-mentioned second female parent,
After 75% ethanol surface sterilization, being placed on division culture medium, under the conditions of 25 DEG C～27 DEG C, lucifuge recovers training
Support one day, after cultivating 3 days under illumination, be transferred in 1/2MS+IBA 4.8mg/L+NAA 0.04mg/L culture medium
Continue to cultivate shading renewal cultivation one day；
S4, grow to more than 1cm to resistant buds after, move it into root media: improvement MS+0.04mg/L
Continuing in NAA+0.75mg/L 6-BA+0.075mg/L IBA+35g/L sucrose medium to cultivate, growth of seedling is extremely
After well developed root system, stem and leaf stalwartness, quality trait is measured comprehensively, screening yield is high, comprehensive resistance is strong,
Colory strain moves in greenhouse, and conventional cultivation, until producing seeds.
The above is only the preferred embodiment of the present invention, it is noted that common for the art
For technical staff, under the premise without departing from the principles of the invention, it is also possible to make some improvements and modifications,
These improvements and modifications also should be regarded as protection scope of the present invention.
Claims (6)
1. the cross breeding method of a high-yield corn, it is characterised in that comprise the steps:
S1, select suitable local plantation, Resistant, the high-quality that anti-fall, tiller is weak, fringe is big, setting percentage is high,
High-yield variety hybridizes as Semen Maydis male parent and Semen Maydis female parent, obtains cross combination F1 generation seed；
S2, under the conditions of selection growth, utilize gained F1 generation seed selfing formed second cycle line colony, and
Therefrom select that there is special strain selfing 10 generation of screening character more than, obtain the second female parent that character is stable；Tool
Body, under the conditions of low-phosphorous, sow seed, between milpa length is to seven leaf phases to emulsified phase, to Semen Maydis
Plant carries out Osmotic treatment, no longer waters in corn-growing regions and pours water；And after milk stage in 1～2 day
In corn-growing regions, pouring saline-alkali water is to saline-alkali water above ground level 1～5cm, normally waters Semen Maydis afterwards and pours water
Point；
S3, will be containing BT insecticidal neurotoxin gene, bar gene, maize dwarf mosaic virus by particle bombardment
The DNA plasmid of coat protein gene MDMVCPDNA imports Seedling cell, tissue or the organ of above-mentioned second female parent,
After 75% ethanol surface sterilization, being placed on division culture medium, under the conditions of 25 DEG C～27 DEG C, lucifuge recovers training
Support one day, after cultivating 3 days under illumination, be transferred in 1/2MS+IBA4.8mg/L+NAA0.04mg/L culture medium
Continue to cultivate shading renewal cultivation one day；
S4, grow to more than 1cm to resistant buds after, move it into root media: improvement MS+0.04mg/L
Continuing in NAA+0.75mg/L6-BA+0.075mg/LIBA+35g/L sucrose medium to cultivate, growth of seedling is extremely
After well developed root system, stem and leaf stalwartness, quality trait is measured comprehensively, screening yield is high, comprehensive resistance is strong,
Colory strain moves in greenhouse, and conventional cultivation, until producing seeds.
The cross breeding method of a kind of high-yield corn the most according to claim 1, it is characterised in that
Every liter of saline-alkali water comprises 23.176 grams of sodium chloride, 3.16 grams of magnesium chloride, 2.148 grams of magnesium sulfate, calcium chloride
2.243 grams, sodium bicarbonate 0.379 gram, 0.623 gram of potassium chloride.
The cross breeding method of a kind of high-yield corn the most according to claim 1, it is characterised in that
Improvement MS in institute's step S4 includes manganese methionine 15mg/L, zinc-amino acid chelate 8mg/L, nucleoside hydrochlorate
13mg/L, magnesium amino acid chelate 6mg/L, Selenomethionine 5.5mg/L, phytase 0.85mg/L, pyridine acid
Chromium 0.065mg/L, calcium iodate 0.3mg/L, lysine 6mg/L, humic acid chelating cobalt 0.065mg/L,
Albumen germanium 0.75mg/L, iodine yeast 6mg/L, thiamine hydrochloride 10mg/L, pyridoxine hydrochloride 3mg/L,
EM original solution 35mg/L, cyclohexanhexanol 27mg/L.
The cross breeding method of a kind of high-yield corn the most according to claim 3, it is characterised in that
5 '-adenylic acid dipotassium that described nucleoside hydrochlorate obtains after including ribonucleic acid basic hydrolysis or sodium, 5 '-bird gland
Thuja acid dipotassium or sodium, 5 '-cytidylic acid dipotassium or sodium and 5 '-uridylic acid dipotassium.
The cross breeding method of a kind of high-yield corn the most according to claim 1, it is characterised in that institute
State screening character include in following character any three kinds:
Corn kernel: long awns, white shell and yellow；
Plant height: 100～110cm；
Fruit ear: tassel row number 10～18 row mass of 1000 kernel 200～300g, fecundity is good；
Fringe position is on the low side: 40～60cm.
The cross breeding method of a kind of high-yield corn the most according to claim 1, it is characterised in that institute
State step S2 prior to seeding, need to be at corn-growing regions applying base fertilizer: nitrogenous fertilizer 110kg N/hm2, phosphate fertilizer 5kg
P2O5/hm2With potash fertilizer 12kg K2O/hm2, to form low-phosphorous condition.
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109042312A (en) * | 2018-10-29 | 2018-12-21 | 山西省农业科学院小麦研究所 | A kind of corn cross breeding method |
| CN110771499A (en) * | 2019-12-12 | 2020-02-11 | 重庆三峡农业科学院(重庆市万州区甘宁蚕种场) | Method for improving seed production yield of waxy corn |
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| CN110771499A (en) * | 2019-12-12 | 2020-02-11 | 重庆三峡农业科学院(重庆市万州区甘宁蚕种场) | Method for improving seed production yield of waxy corn |
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Application publication date: 20161012 |