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CN105165602A - Method for breeding glyphosate-resistant/tolerant novel germplasm by utilizing wild soybeans - Google Patents

Method for breeding glyphosate-resistant/tolerant novel germplasm by utilizing wild soybeans Download PDF

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CN105165602A
CN105165602A CN201510676655.5A CN201510676655A CN105165602A CN 105165602 A CN105165602 A CN 105165602A CN 201510676655 A CN201510676655 A CN 201510676655A CN 105165602 A CN105165602 A CN 105165602A
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glyphosate
wild soybean
resistance
wild
soybean
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CN105165602B (en
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毕影东
来永才
陶波
陈受宜
李炜
张劲松
刘明
张必弦
贺超英
张万科
刘淼
李琬
肖佳雷
王燕平
王萍
李擎天
郭东林
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Institute of Tillage and Cultivation Heilongjiang Academy of Agricultural Sciences
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Institute of Tillage and Cultivation Heilongjiang Academy of Agricultural Sciences
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Abstract

The invention belongs to the field of utilization and research of mild plant resources, and particularly relates to a method for breeding a glyphosate-resistant/tolerant novel germplasm by utilizing wild soybeans. The method for breeding the glyphosate-resistant/tolerant novel germplasm utilizes different concentrations of glyphosate to treat the wild soybeans at the seedling stage to screen out wild soybeans with resistance/tolerance to the glyphosate, and then utilizes the screened wild soybeans with resistance/tolerance to carry out interspecific hybridization with cultivated soybeans, so that an interspecific hybridization generation is obtained, glyphosate resistance/tolerance screening is carried out on the selfing line of the interspecific hybridization generation, and in combination with phenotypic screening, the method breeds out the novel germplasm with resistance/tolerance to the glyphosate. The invention determines a wild soybean resource resistance standard evaluation system, and creates the method for screening the wild soybeans with resistance/tolerance, moreover, by utilizing the wild soybeans with resistance/tolerance to carry out interspecific hybridization, the invention creates the method for breeding the glyphosate-resistant/tolerant novel germplasm, and the method is simple, practical and highly efficient, and is also applicable to the research of the other characters of the wild soybeans.

Description

A kind ofly wild soybean seed selection is utilized to resist/the method for glyphosate tolerant new germ plasm
Technical field
The invention belongs to wild plant resource research on utilization field, be specifically related to a kind ofly utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm.
Background technology
Glyphosate is a kind of very important organic phosphates weed killer herbicide, and it a kind ofly imposes on blade, wide spectrum, nonselective weed killer herbicide, has extremely strong control ability for annual and perennial weed.
Proceed to Antiglyphosate gene in genetically engineered soybean, make the susceptibility of genetically engineered soybean to glyphosate lower, the shikimic acid pathway in Transgenic soybean plants can normally carry out, thus shows the resistance to glyphosate.From genetically engineered soybean in 1994 since the U.S. gets permission to commercially produce, the cultivated area of global genetically engineered soybean rapidly increases.But, due to the establishing in large scale of resistance glyphosate genetically engineered soybean, and long-term, the single use of glyphosate, cause the frequency of occurrences of field resistance glyphosate weeds greatly to rise; And research finds, when being subject to the injury of damage by disease and insect and excessive rainfall or other weed killer herbicide, Transgenic soybean plants can not decompose glyphosate rapidly, thus occur symptom of chemical damage; Have the resistance to life of some kinds to the sweet unicorn of grass lower than other kinds in the screening process of simultaneously resistant variety, thus cause genetically engineered soybean to occur drug injury problem after spraying glyphosate.
Wild soybean is the original ancestry kind of the cultivated soybean, the wild soybean grown under natural conditions, the artificial impact selected is not subject at genetic aspect, therefore a lot of abundant functional gene is not lost because of artificial selection, these genes may be in Soybean production in the urgent need to key gene, high protein in Wild Soybean Resources, resist more, wide adaptability and the high excellent material of reproduction coefficient can be directly used in the cultivated soybean high-quality, disease-resistant, SOYBEAN IN HIGH-YIELD BREEDING research, the genetic diversity of Wild Soybean Resources is significant to realizing the cultivated soybean genetic improvement.(Wang Kejing, Li Fushan; China's wild soybean (G.soja) germ plasm resource and germplasm innovation thereof utilize, Chinese agriculture science and technology Leader, 2000,6 (2): 69-72: Tian Qingzhen, Gai Junyi; The study and utilization of Wild Soybean Germplasms, plant genetic resources science, 2000,4 (1): 61-65)
The defect existed based on resistance glyphosate genetically engineered soybean and the genetic diversity of Wild Soybean Resources, screening occurring in nature has the Wild Soybean Resources of resistance glyphosate and utilizes this resource to carry out quality-improving just to seem particularly important.
Summary of the invention
The object of the invention is to utilize there is glyphosate herbicidal resist/wild resource of patience improves existing the cultivated soybean kind, the invention provides screening, qualification has glyphosate herbicidal and resists/method of the wild soybean plant of patience, and utilize the resource screened to carry out the new germ plasm that interspecific cross seed selection has anti-/ glyphosate tolerant.
Technical scheme of the present invention:
A kind ofly wild soybean seed selection is utilized to resist/the method for glyphosate tolerant new germ plasm, described seed selection resists/and the method for glyphosate tolerant new germ plasm utilizes the glyphosate process wild soybean in seedling stage of variable concentrations gradient to filter out wild soybean glyphosate to anti-/ patience, then the wild soybean with anti-/ patience filtered out and the cultivated soybean is utilized to carry out interspecific cross, obtain Interspecific Hybrids, to the inbred line of described Interspecific Hybrids carry out glyphosate resist/patience screening select new germ plasm glyphosate to anti-/ patience in conjunction with phenotypic screen.
Further, described seed selection resist/method of glyphosate tolerant new germ plasm comprises the steps:
(1) process before wild soybean sowing to be identified: central back to sub-navel place at the seed kernel of wild soybean to be identified, remove the seed coat of 2mm × 2mm size, it is stand-by that the seed after process puts into 3-6 DEG C of refrigerator;
(2) wild soybean is to the qualification of glyphosate resistance: the wild soybean to be identified after plantation step (1) process, adopt the described wild soybean to be identified of the method for field test qualification to glyphosate anti-/ resistance, 10-15 glyphosate concentration for the treatment of is set, each described glyphosate concentration for the treatment of correspondence arranges an experimental plot, described wild soybean to be identified is planted in each described experimental plot, described wild soybean hole pressing plantation to be identified, repeating groups and blank group are set, when the two panels trifoliolate leaf of the wild soybean of plantation launches completely, evenly glyphosate is sprayed to described two panels trifoliolate leaf,
(3) evaluation of wild soybean glyphosate resistance rank: the phenotype of the wild soybean plant after spray medicine was observed in the 7th day after step (2) process, the object of described Phenotypic Observation is the area ratio of dead leaf, and according to viewed phenotype determination wild soybean to the resistance rank of glyphosate, and the resistance rank of wild soybean to be identified described under different disposal concentration is added up;
(4) determination of Resistance resource: the wild soybean resistance rank data described to be identified obtained according to step (3), carries out Frequency statistics and variance analysis to determine screening the optimal glyphosate concentration for the treatment of of anti-/ glyphosate tolerant wild soybean; Multiple comparison analyse is carried out to the resistance of the wild soybean all to be identified under described optimal glyphosate concentration for the treatment of, with find out to glyphosate anti-/ wild soybean of Nai Sheng significant difference compared with other wild soybeans, the Wild Soybean Resources of the described resistance to glyphosate/patience significant difference is defined as the wild soybean with anti-/ patience
(5) interspecific cross initiative new germ plasm: selecting Hefeng47 as female parent, is that male parent is cross-breeding to filter out the wild soybean with anti-/ resistance to life in step (4), obtains F0 generation hybridization grain, collects hybridization grain and carry out preservation;
(6) Interspecific Hybrids glyphosate resistance, phenotypic screen: continue the F0 generation hybridization grain that plantation step (5) is gathered in the crops, to self progeny F2 until F5 carries out glyphosate resistance screening by step (1) to step (5) described method, and proceed self propagated in conjunction with offspring's phenotypic screen superior progeny, according to offspring between significance analysis determination good species, filter out anti-glyphosate germplasm.
Further, the glyphosate sprayed described in step (2) is 41% glyphosate isopropyl salt aqua.
Further, step (2) wild soybean, in the qualification of glyphosate resistance, arranges 10 glyphosate concentration for the treatment of, 10 glyphosate concentration for the treatment of of described setting are respectively 0ml/ mu, 25ml/ mu, 50ml/ mu, 100ml/ mu, 125ml/ mu, 150ml/ mu, 175ml/ mu, 200ml/ mu, 225ml/ mu, 250ml/ mu.
Further, set up resistance standard evaluation system according to the resistance level method for distinguishing of phenotype determination wild soybean to glyphosate described in step (3), described resistance standard evaluation system is divided into according to the resistance rank of dead leaf area ratio by wild soybean to be identified: 0 grade, 1 grade, 2 grades, 3 grades, 4 grades and 5 grades, and the described resistance rank dead leaf area ratio corresponding from 0 grade to 5 grades is: 0-10.0%, 10.1-20%, 20.1-40%, 40.1-70%, 70.1-90%, 90.1-100.0%.
Further, selecting No. 51, black agriculture as maternal in step (5), is that male parent is cross-breeding with the resistance Wild Soybean Resources filtered out in step (4), obtains F0 generation hybridization grain, collects hybridization grain and carry out preservation.
Further, in step (5) interspecific cross initiative new germ plasm process, multi-functional pollinating tool bag and soybean pollination emasculation device is adopted to carry out.
Beneficial effect of the present invention:
The present invention carries out part process to wild soybean seed coat, guarantees to emerge unanimously; The present invention utilizes glyphosate isopropyl salt (41% aqua) to process screening Wild Soybean Resources, and by arranging multiple glyphosate concentration for the treatment of, understanding the resistant range of Wild Soybean Resources to glyphosate resistance in depth, accurately screening Resistance resource; Determine the evaluation criterion that accurately divides glyphosate resistance rank, be divided into 6 resistance ranks and assignment according to the character mutation of wild soybean poisoning, according to the resource of variance analysis determination significant difference, thus greatly strengthen reliability; Adopt the Multi-functional soybean pollinating tool bag (grant number Zl20140182009.4) of our unit and soybean pollination emasculation device (grant number Zl201420830748.x) to carry out the hybridization of wild soybean and the cultivated soybean, improve the success rate of hybridization.
The present invention determines Wild Soybean Resources resistance standard evaluation system, create and carry out interspecific cross seed selection and resist/the method for glyphosate tolerant new germ plasm from wild resource Evaluating and screening to utilizing the wild soybean with anti-/ patience of screening, the method is simple, practical, efficient, is also applicable to the research of other proterties of wild soybean.
Accompanying drawing explanation
Fig. 1. the method schematic diagram of the screening and assessment of anti-/ glyphosate tolerant wild soybean and new germ plasm initiative;
Fig. 2. wild soybean seed coat process schematic diagram;
Fig. 3. the phenotype schematic diagram of 6 resistance rank wild soybeans after the process of wild soybean glyphosate in seedling stage;
Fig. 4. to resistance variation diagram under the Frequency statistics analysis chart of Wild Soybean Resources resistance rank and variable concentrations.
Embodiment
In order to make object of the present invention, technical scheme and advantage clearly understand, below in conjunction with drawings and Examples, the present invention is explained in further detail.Should be appreciated that specific embodiment described herein only for explaining the present invention, being not intended to limit the present invention.
On the contrary, the present invention is contained any by the substituting of making on marrow of the present invention and scope of defining of claim, amendment, equivalent method and scheme.Further, in order to make the public have a better understanding to the present invention, in hereafter details of the present invention being described, detailedly describe some specific detail sections.Do not have the description of these detail sections can understand the present invention completely for a person skilled in the art yet.
Embodiment 1
A kind ofly wild soybean seed selection is utilized to resist/the method for glyphosate tolerant new germ plasm, described seed selection resists/and the method for glyphosate tolerant new germ plasm utilizes the glyphosate process wild soybean in seedling stage of variable concentrations gradient to filter out wild soybean glyphosate to anti-/ patience, then the wild soybean with anti-/ patience filtered out and the cultivated soybean is utilized to carry out interspecific cross, obtain Interspecific Hybrids, to the inbred line of described Interspecific Hybrids carry out glyphosate resist/patience screening select the new germ plasm (as shown in Figure 1) glyphosate to anti-/ resistance to life in conjunction with phenotypic screen.
Described seed selection resists/and the method for glyphosate tolerant new germ plasm comprises the steps:
(1) process (as shown in Figure 2) before wild soybean sowing to be identified: because wild soybean will protect seed under severe conditions and in barren environment; allow them keep resting state until there is being applicable to the condition of rudiment always, thus realize long-term existence.The hard permeability of its seed coat is poor, for making all qualification resource growth synchronously be convenient to Identification, need process, guarantee that sprouting is emerged unanimously to wild soybean.Every part of resource chooses 20, the evenly full seed of seed, in seed central authorities back to sub-navel, removes the seed coat of about 2mm × 2mm size, and noting can not the embryo of seed, puts into 3-6 DEG C of Refrigerator store stand-by after process;
(2) wild soybean is to the qualification of glyphosate resistance:
Wild soybean to be identified after plantation step (1) process, adopt the described wild soybean to be identified of the method for field test qualification to glyphosate anti-/ resistance, 10 glyphosate concentration for the treatment of are set, each described glyphosate concentration for the treatment of correspondence arranges an experimental plot, described wild soybean to be identified is planted in each described experimental plot, described wild soybean hole pressing plantation to be identified, five, every cave, final singling one strain after emerging, repeat for three times, if blank, when the two panels trifoliolate leaf of the wild soybean of plantation launches completely, evenly glyphosate is sprayed to described two panels trifoliolate leaf,
(3) evaluation of wild soybean glyphosate resistance rank: the 7th day after step (2) the process phenotype to the wild soybean plant after spray medicine and symptom of chemical damage are observed, the resistance rank of wild soybean to glyphosate is determined in conjunction with resistance standard evaluation system according to viewed phenotype, and the resistance rank (as shown in table 1, Fig. 3) of different described wild soybean to be identified under adding up different disposal concentration;
Table 1 wild soybean resists/patience evaluation criterion to the resistance rank of glyphosate and to glyphosate
(4) determination of Resistance resource: the wild soybean resistance rank data described to be identified obtained according to step (3), spss22 statistical analysis software is utilized to carry out Frequency statistics and variance analysis, number statistical analysis frequently (as shown in Figure 4) shows: resistance rank is that normal distribution illustrates that the classification of resistance rank is reasonable, variance analysis (as shown in table 2) shows: remarkable to the Resistant Difference of glyphosate between different disposal concentration, further analysis shows when spraying concentration is 175ml/ mu, resistance index change is extremely remarkable, show to adopt this concentration for the treatment of to screen to resist/glyphosate tolerant Wild Soybean Resources is comparatively reasonable, be that the resistance of 22 portions of wild soybeans of 175ml/ mu carries out multiple comparison analyse to spraying concentration, Duncan inspection is adopted to show No. 4 wild soybean resistances significant difference compared with other wild soybeans, average resistance index is 2.66, adopt above-mentioned same method again the duplicate test of antagonism plant show that this resource resistance is remarkable, is defined as Resistance resource (as shown in table 3) by No. 4 Wild Soybean Resourcess,
Significant difference analysis between table 2 variable concentrations
b.Dunnettt-teststreatonegroupasacontrol,andcompareallothergroupsagainstit.
Significant difference analysis between table 3 matter not of the same race
Resistance index
Duncan a
Meansforgroupsinhomogeneous
subsetsaredisplayed
(5) interspecific cross initiative new germ plasm: this patent is for guaranteeing that sprouting is emerged unanimously.Process wild soybean before sowing, the same step of processing method (1), plant in experimental field at the beginning of 5 months, seeding row spacing 10cm × 40cm, executes a small amount of ash in cave, use bamboo built at the beginning of 5 months.Guarantee late June and the cultivated soybean flower synchronization, because wild soybean flower pattern is less, the flower that defloration lobe is not opened, keel not yet split be selected to adopt powder.Multi-functional soybean pollinating tool bag (grant number Zl20140182009.4) is utilized to carry out, Comprehensive Traits with reference to Soybean Germplasm is selected to close rich 47 as maternal, select the Wild Soybean Resources 1 part of (high resistance of the resistance glyphosate weed killer herbicide filtered out, 01-680) be male parent, after bud is selected in maternal middle and upper part, remove other without bud, bud bottom is gently pinched with have gentle hands, to tear the peripheral sepal of bud with tweezers, then the crown portion exposed is pinched with tweezers middle front part, tendency vexil direction proposes gently, is taken out together by flower pesticide with corolla.After determining that flower pesticide all takes out, with tweezers tip by the pollen silk pinch off under She or taking-up, whether intactly observe stigma and style.The wild soybean flower that defloration lobe is not opened, keel not yet split is selected to adopt powder.Extractd by the sepal gathering flower, strip off petal, exposes flower pesticide; flower pesticide is smeared pollination lightly on column cap, wraps up whole emasculation flower with fresh blade again after pollination, and buckle well isolation with petiole; so both play insulation blocking effect, certain humidity can be kept again, hang up properly label.Maternal should selection does not have the bud in autophilous growth acme to be advisable, and namely sepal is about to split, and can see the color of corolla, and corolla does not exceed sepal height.When removing sepal and corolla, flower pesticide is fitly arranged in around gynoecium in white yellow, graininess, after contact, does not have pollen to come off; Male parent should select sepal to split completely, can see the color of corolla, and corolla does not have crannied bud.Be easy to act as most when parent enters full-bloom stage and bud more period and start hybridization, can hybridization time be saved like this, improve survival rate high.Select emasculation in the morning, directly this Crossing system of pollination, generally start emasculation in the morning, gather paternal pollen simultaneously, pollinate, noon, crossing work completed substantially; After pollination, the anthocaulus of hybridization flower hangs up plastics board, writes hybrid combination code name or title, pollination date and operator's name exactly, and perform record on working copy.After pollinating 1 week, remove the blade that parcel is taken in hybridization, as hybridized successfully, then ovary has started to expand, and checks that plucking hybridization spends the other bud newly grown.Being hybridized to pod rate for improving, should strengthening water management, as arid, hybridization garden will be irrigated in time, and keep soil moisture sufficient, field microclimate is moistening.After maturation, by combination results hybridization in time beanpod, after drying threshing, the hybrid seed of same combination is loaded in seed packet together with plastics board, properly preserves;
(6) Interspecific Hybrids glyphosate resistance, phenotypic screen: in step (5), F0 generation hybridization grain is being obtained then at the beginning of 10 months, continue the F0 generation hybridization grain of plantation results, to self progeny F2 until F5 carries out glyphosate resistance screening by step (1) to step (5) described method, and proceed self propagated in conjunction with offspring's phenotypic screen superior progeny, according to offspring between significance analysis determination good species, filter out anti-glyphosate germplasm.
The glyphosate sprayed described in step (2) is 41% glyphosate isopropyl salt aqua.
Step (2) wild soybean is in the qualification of glyphosate resistance, arrange 10 glyphosate concentration for the treatment of, 10 glyphosate concentration for the treatment of of described setting are respectively 0ml/ mu, 25ml/ mu, 50ml/ mu, 100ml/ mu, 125ml/ mu, 150ml/ mu, 175ml/ mu, 200ml/ mu, 225ml/ mu, 250ml/ mu.
Selecting black agriculture 51 as female parent in step (5), is that male parent is cross-breeding with the resistance Wild Soybean Resources filtered out in step (4), obtains F0 generation hybridization grain, collects hybridization grain and carry out preservation.

Claims (7)

1. one kind utilizes wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, described seed selection resists/and the method for glyphosate tolerant new germ plasm utilizes the glyphosate process wild soybean in seedling stage of variable concentrations gradient to filter out wild soybean glyphosate to anti-/ patience, then the wild soybean with anti-/ patience filtered out and the cultivated soybean is utilized to carry out interspecific cross, obtain Interspecific Hybrids, to the inbred line of described Interspecific Hybrids carry out glyphosate resist/patience screening select new germ plasm glyphosate to anti-/ patience in conjunction with phenotypic screen.
2. a kind ofly according to claim 1 utilizing wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, described seed selection resists/and the method for glyphosate tolerant new germ plasm comprises the steps:
(1) process before wild soybean sowing to be identified: central back to sub-navel place at the seed kernel of wild soybean to be identified, remove the seed coat of 2mm × 2mm size, it is stand-by that the seed after process puts into 3-6 DEG C of refrigerator;
(2) wild soybean is to the qualification of glyphosate resistance: the wild soybean to be identified after plantation step (1) process, adopt the described wild soybean to be identified of the method for field test qualification to glyphosate anti-/ resistance, 10-15 glyphosate concentration for the treatment of is set, each described glyphosate concentration for the treatment of correspondence arranges an experimental plot, described wild soybean to be identified is planted in each described experimental plot, described wild soybean hole pressing plantation to be identified, repeating groups and blank group are set, when the two panels trifoliolate leaf of the wild soybean of plantation launches completely, evenly glyphosate is sprayed to described two panels trifoliolate leaf,
(3) evaluation of wild soybean glyphosate resistance rank: the phenotype of the wild soybean plant after spray medicine was observed in the 7th day after step (2) process, the object of described Phenotypic Observation is the area ratio of dead leaf, and according to viewed phenotype determination wild soybean to the resistance rank of glyphosate, and the resistance rank of wild soybean to be identified described under different disposal concentration is added up;
(4) determination of Resistance resource: the wild soybean resistance rank data described to be identified obtained according to step (3), carries out Frequency statistics and variance analysis to determine screening the optimal glyphosate concentration for the treatment of of anti-/ glyphosate tolerant wild soybean; Multiple comparison analyse is carried out to the resistance of the wild soybean all to be identified under described optimal glyphosate concentration for the treatment of, with find out to glyphosate anti-/ wild soybean of patience significant difference compared with other wild soybeans, the Wild Soybean Resources of the described resistance to glyphosate/patience significant difference is defined as the wild soybean with anti-/ patience
(5) interspecific cross initiative new germ plasm: selecting Hefeng47 as female parent, is that male parent is cross-breeding to filter out the wild soybean with anti-/ patience in step (4), obtains F0 generation hybridization grain, collects hybridization grain and carry out preservation;
(6) Interspecific Hybrids glyphosate resistance, phenotypic screen: continue the F0 generation hybridization grain that plantation step (5) is gathered in the crops, to self progeny F2 until F5 carries out glyphosate resistance screening by step (1) to step (5) described method, and proceed self propagated in conjunction with offspring's phenotypic screen superior progeny, according to offspring between significance analysis determination good species, filter out anti-glyphosate germplasm.
3. a kind ofly according to claim 2 utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, the glyphosate sprayed described in step (2) is 41% glyphosate isopropyl salt aqua.
4. a kind ofly according to claim 2 utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, step (2) wild soybean is in the qualification of glyphosate resistance, arrange 10 glyphosate concentration for the treatment of, 10 glyphosate concentration for the treatment of of described setting are respectively 0ml/ mu, 25ml/ mu, 50ml/ mu, 100ml/ mu, 125ml/ mu, 150ml/ mu, 175ml/ mu, 200ml/ mu, 225ml/ mu, 250ml/ mu.
5. a kind ofly according to claim 2 utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, set up resistance standard evaluation system according to the resistance level method for distinguishing of phenotype determination wild soybean to glyphosate described in step (3), described resistance standard evaluation system is divided into according to the resistance rank of dead leaf area ratio by wild soybean to be identified: 0 grade, 1 grade, 2 grades, 3 grades, 4 grades and 5 grades, the described resistance rank dead leaf area ratio corresponding from 0 grade to 5 grades is: 0-10.0%, 10.1-20%, 20.1-40%, 40.1-70%, 70.1-90%, 90.1-100.0%.
6. a kind ofly according to claim 2 utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, select No. 51, black agriculture as maternal in step (5), be that male parent is cross-breeding with the resistance Wild Soybean Resources filtered out in step (4), obtain F0 generation hybridization grain, collect hybridization grain and carry out preservation.
7. a kind ofly according to claim 2 utilize wild soybean seed selection to resist the/method of glyphosate tolerant new germ plasm, it is characterized in that, in step (5) interspecific cross initiative new germ plasm process, adopt multi-functional pollinating tool bag and soybean pollination emasculation device to carry out.
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CN114467743A (en) * 2022-03-02 2022-05-13 黑龙江省农业科学院耕作栽培研究所 Method for creating new germplasm of soybean with phytophthora root rot resistance

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