CN106489716A - A kind of Rice Resistance linum leaf spot breeding method - Google Patents
A kind of Rice Resistance linum leaf spot breeding method Download PDFInfo
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Abstract
The invention discloses as follows the step of a kind of Rice Resistance linum leaf spot breeding method, the breeding method：（1）Select japonica rice material to be improved and the anti-source of water rice brown spot long-grained nonglutinous rice；（2）The identification that Raw toxin patience concentration is carried out to the japonica rice material to be improved for selecting；（3）Hybridization obtains three and hands over F1 cenospecies；（4）The F1 cenospecies row of carrying out Xian culture medium Anther Cultures again are handed over to three；（5）Bloom control individual plant sowing obtains flower training family, and further economical character identification, linum leaf spot resistance screening obtain Keng rice lines.Linum leaf spot genetics of resistance material in long-grained nonglutinous rice can efficiently and exclusively be penetrated into japonica rice by the breeding method of the present invention, Rice Resistance linum leaf spot breeding can be completed within 3 years or so, the linum leaf spot breeding for disease resistance cycle is greatly shortened, is worth with larger Breeding Application.
Description
Technical field
The present invention relates to a kind of Rice Resistance linum leaf spot breeding method, belongs to agricultural high-tech field.
Background technology
Oryza sativa L. is one of important cereal crops of China, and recent years, one kind was by a kind of microbial rice fungus diseases evil of rice Bipolaris -- water rice brown spot（Rice brown spot）In the common generation in many places of China.Water rice brown spot（Rice
brown spot）Also known as Hemintho-sporum leaf spot of rice plants, water helminthosporium leaf spot of rice, it is by a kind of microbial global fungal disease of Folium Sesami speckle.In recent years, linum leaf spot generally occurs in many places of China, such as Guangxi Yongning county, Guangxi Zhaoping County, Tianjin, one band of Tangshan and Qinhuangdao, coastal rice regions of Hebei province Ji Dong etc., generally result in the 10%-30% underproduction, severe patient more than 50%, even almost have no harvest, very big economic loss is caused to peasant, the serious contusion kind grain enthusiasm of peasant brings safely serious challenge to the rice industry of country.
Water rice brown spot can be fallen ill to the period of maturation from rice shoot in Oryza sativa L., and disease symptom is showed based on blade, be secondly stem, fringe portion, grain, also cause harm once in a while stipes and sheath.It is the small-sized speckle of brown at the beginning of scab on leaf, gradually expand ovalisation, circle, spindle, fusiformis etc. afterwards, surrounding brown, dark brown, central authorities are greyish white, and edge carries yellow halo, but without the downright bad line extended along leaf margin, when being in a bad way, on blade, scab merges to form irregular big speckle, turns yellow to withered.Fringe portion takes place mostly in fringe neck, brown or taupe, similar to rice blast symptom, and when moist, sick portion produces the mould layer of Lycoperdon polymorphum Vitt, i.e. conidium.When grain morbidity is more early, in grey black, in base, rice grain is changed into Lycoperdon polymorphum Vitt or becomes blighted grain, and when moist, seed surface can produce a large amount of black mustiness things, and the later stage, aggrieved scab was little, and edge is not obvious, and seriously ill grain matter is crisp frangible.
The color modest grade of Wu in 2008 has investigated Zhaoping County evening rice brown spot field occurrence characteristic, according to field investigation, there is different degrees of prevalence in Zhaoping County in this disease, particularly in the Oryza sativa L. booting later stage, whole county occurring area accounts for the 37.50% of Monitoring of Paddy Rice Plant Area, cause harm serious, production loss is big, and in severe disease area disease leaf incidence rate up to 100%, disease index is up to 78.32, the lighter field of morbidity, typically 15% or so, serious field, loss rate reach 30% or so to loss rate, the even complete whole strain in field of some fields is withered, and No kernels or seeds are gathered, as in a year of scarcity.
Water rice brown spot pathogen conidiophore list life 2-4 networks of roots are given birth to, not branch, and in knee complications, brown or dark brown, barrier film 4-16 is individual, 99-345 × 4-11 μm on top.The conidium oblong fusiformis shape of falling club, honest or bend to a side, brown, two ends be gradually narrow, blunt circle, 3-11 barrier film, 14.4-19.2 μm of 24.2-81.8 μ ms.Spore germination two-stage is sprouted, and observes from the detached rice Bipolaris sacchari of different places, and its form is essentially identical but variant, and the water rice brown spot pathogen for reflecting different regions source belongs to different biological strains.
Folium Sesami plaque is attached on seed rice or diseased plant residuum with conidium or mycelium survives the winter, and becomes next year primary source of infection.Conidium can survive 2-3 in drying condition, and mycelium can be survived 3-4.The pathogen that survives the winter can directly encroach on seedling, form conidium, cause rice seedling bed or Honda primary infection, under appropriate conditions spore germination to produce scab through directly invading or invading through pore, form conidium immediately and infected again by gas biography, hearsay etc..General Rice Seedlings are most easily susceptible, but disease is mainly manifested in rice plant of tillering stage to heading stage.General fertilizer deficiency or barren field, potassium deficiency fertilizer, soil are the field of the serious field of acid or sand leak, hydropenia or long-term hydrops, weight of falling ill.Linum leaf spot symptom can all be fallen ill from seedling stage to harvest time, and pathogenic bacteria can endanger each position of plant above ground, occur with blade most universal.
Linum leaf spot has many reasons, such as environment, weather, soil, water and fertilizer management, rice varieties etc..Water and fertilizer management is in close relations with linum leaf spot, and it is improper to pour water, and rice terrace keeps Deep irrigation for a long time, root system is caused to be in anaerobic state, the bad major incentive that often linum leaf spot occurs of rice nutrition is made, nitrogenous fertilizer, siliceous fertilizer, potash fertilizer are very few, then favourable disease occurs.At present in rice varieties, there is no very big to linum leaf spot Resistant Difference between immunological species, but different cultivars, in general, japonica rice is more susceptible than long-grained nonglutinous rice.
The prophylactico-therapeutic measuress of linum leaf spot should take the plant protection policy of " putting prevention first; integrated control " according to the rule of its occurrence and development, may be without positive effect if relying on single prophylactico-therapeutic measuress, or causing more serious consequence, the such as single antibacterial of life-time service easily makes pathogen develop immunity to drugs, in integrated control, attached most importance to cultural control, according to specific time and site, with multiple prophylactico-therapeutic measuress such as chemical prevention, physical control, Biological control.Prevention and controls mainly improve cropping system and chemical prevention at present, once occur, it is impossible to eradicate, and pollute environment, cause pesticide residues, give people class health and bring harm.
For a long time, chemical agent and cultivation step are depended on to the preventing and treating of water rice brown spot, but prevention effect is not satisfactory.Integrated control technique is taken to prevent the generation of linum leaf spot in agricultural production now, Main Means are to take some preventive measures, Drug therapy is carried out such as from disease-resistant variety, on cultivating using antibiotic and sterilizing means and applying pesticides, and set up linum leaf spot forecast prognoses system, long-term production practices prove that the selection-breeding and application of water rice brown spot resistant variety are to prevent and treat the most economical effective measures of banded sclerotial blight.
In the early stage, breeding is carried out using traditional disease resistant and breeding method mainly in anti-linum leaf spot breeding, such as using introducing a fine variety, the conventional breeding approach such as selection and use, cross-breeding select a collection of with vertical resistance and the preferable excellent strain of horizontal resistance.But as large-area unification uses certain resistance gene, the orthoselection to linum leaf spot colony is caused to act on, the complexity and pathogenic multiformity of linum leaf spot Biological Strains of The Pest heredity causes the composition of the virulence gene of linum leaf spot pathogenic bacteria colony to change, often the resistant variety of selection-breeding produces the dominant races that can infect the kind after 3-5 is promoted, and the resistance for ultimately resulting in new varieties is lost.Conventional breeding elapsed time is long, and especially as the increase of linum leaf spot pathogenic bacteria mutation frequency, the speed of a new varieties incubation has not caught up with the mutating speed of linum leaf spot pathogenic bacteria.
Indica rice differentiation and significant difference deeply, them are made to differ from one another at aspects such as yield, quality and resistance, one of them Resistant Difference highly significant in linum leaf spot pathogenic bacteria, the multipair linum leaf spot of rice variety has higher resistance level, and japonica rice is poor to the resistance of linum leaf spot, show that long-grained nonglutinous rice genome is contained within the unexistent anti-linum leaf spot gene of a large amount of japonica rice and exists.If the anti-linum leaf spot gene in long-grained nonglutinous rice is transferred into the anti-linum leaf spot japonica rice variety of japonica rice genome culture by accurate efficient means, there is in production high value.
With the development of biotechnology, many new breeding techniques are occurred in that again in addition to conventional cross-breeding, this give Rice Resistance linum leaf spot breeding new way, one kind is to extract Raw toxin using linum leaf spot culture fluid alternatively to press, screening mutant is carried out on a cellular level, and cultivation new disease-resistant varieties is combined with traditional breeding method, but low by the frequency of Raw toxin induced mutationss, screening amount is very big；It is exactly for second the molecular breeding technology with transgenic technology and molecular marker assisted selection breeding technique as representative.The appearance of molecular breeding technology, it was a kind of feasible solution originally that breeding difficulty is handed over to Rice Resistance linum leaf spot breeding and Xian round-grained rice, at present, transgenic breeding and the new road that molecular marker assisted selection equimolecular technology Breeding is current agricultural development, the appearance for being known as the agriculture Green revolution of third time by scientific circles.But, the transgenic breeding that anti-linum leaf spot gene and other excellent genes are carried out to specific excellent main breed, originally the improvement that Xian round-grained rice is handed over can be accurately realized, but was resisted due to current transgenic breeding, and molecular marker assisted selection breeding was firstly the need of gene mapping, need also exist for the hybridization backcrossing of generation more than land for growing field crops, can not solve the problems, such as that Xian round-grained rice is handed over, in addition, molecular engineering breeding, all there is a problem of that costly, technical threshold is high, it is difficult to grasped by general breeder.
Content of the invention
The purpose of the present invention is that have the shortcomings that breeding cycle length, screening efficiency are low in present Rice Resistance linum leaf spot conventional breeding, there is provided a kind of can be by linum leaf spot genetics of resistance material efficiently and exclusively penetrates into japonica rice in long-grained nonglutinous rice, breeding cycle is greatly shortened Rice Resistance linum leaf spot breeding method.
The purpose of the present invention is solved by the following technical programs：
A kind of Rice Resistance linum leaf spot breeding method, it is characterised in that：The step of breeding method, is as follows：
（1）Select japonica rice material to be improved and the anti-source of water rice brown spot long-grained nonglutinous rice：Choose that economical character is good, water rice brown spot resistance needs the japonica rice material improved and the japonica rice flower pesticide is seeded to inducing culture in row's Xian culture medium, used as japonica rice material improve, selection economical character is good and the long-grained nonglutinous rice material of anti-linum leaf spot is used as the anti-source of linum leaf spot long-grained nonglutinous rice for japonica rice material of the final choice induction of anther callus rate higher than 35%；
（2）The identification that Raw toxin patience concentration is carried out to the japonica rice material to be improved for selecting：The Extracted toxin liquid of certain gradient concentration is added in division culture medium respectively, to step（1）In the anther callus that obtain of japonica rice material to be improved induction carry out differentiation culture, and select the Extracted toxin liquid of suitable screening pressure level concentration to select pressure as japonica rice Anther Culture Raw toxin；
（3）Hybridization obtains three and hands over F1 cenospecies：Anti- for the linum leaf spot long-grained nonglutinous rice of field planting source is built bridge with wide affinitive material Xian round-grained rice and obtains Hybrid F1, the flower pesticide for choosing Hybrid F1 hands over F1 with japonica rice material to be improved hybridization acquisition cenospecies three again；
（4）The F1 cenospecies row of carrying out Xian culture medium Anther Cultures again are handed over to three：Three hand over the positive season sowing of F1 cenospecies, choosing healthy and strong full flower pesticide carries out arranging Xian culture medium flower pesticide inducing culture, obtain anther callus, and select the anther callus of 0.6～0.9cm of diameter be inoculated in the addition of select pressure concentration Raw toxin division culture medium on, differentiation culture under conditions of 27～29 DEG C of temperature, the dark 10h of light 14h/, obtain flower training seedling, transfer after bloom control height of seedling reaches 3cm successive transfer culture is carried out in root media, strong sprout in transplanting to Turnover Box after bloom control height of seedling reaches 8cm, was transplanted into land for growing field crops after 10～15 days；
（5）Bloom control individual plant sowing obtains flower training family, and further economical character identification, linum leaf spot resistance screening obtain Keng rice lines：Field planting bloom control, the japonica rice training family that individual plant sowing acquisition linum leaf spot resistance obtains improveing is some, cell plantation in the further land for growing field crops of training family of not suiting, with japonica rice material parent to be improved as control, screen agronomic shape well and linum leaf spot resistance obtains the Keng rice lines of improvement.
The step（1）In economical character is good, the japonica rice material that water rice brown spot resistance needs to be improved refer to yield height, good grain quality, linum leaf spot resistance need to be improved and plant height should not too high, period of duration precocious or in ripe japonica rice material；The anti-source of described water rice brown spot long-grained nonglutinous rice refers to yield height, good grain quality, the long-grained nonglutinous rice material of high anti-linum leaf spot.
The step（1）The process that middle selection economical character is good, water rice brown spot resistance needs the japonica rice material that is improved and the japonica rice flower pesticide is seeded to inducing culture in row's Xian culture medium is as follows：
（1.1）Draw materials and pretreatment：The japonica rice material land for growing field crops conventional cultivation that will be chosen, is not broken with rice bud, sword-like leave pulvinus accounts for the 1/3～1/2 of clever shell away from the flower pesticide position as in the middle part of 5～8cm, spike of rice and takes fringe for standard, 2～3 blades of reservation, is taken the fringe time and is typically chosen in the fine day morning 9:00～12:00, after drawing materials, with 70% ethanol surface sterilization, then plastic sheeting encases whole spike of rice, tightens plastic bag mouth, keeps spike of rice moistening, is finally put in refrigerator, Cold pretreatment 3 days at 4 DEG C；
（1.2）Inoculation：After pretreatment, cut rice bud, rice bud is soaked in 3～5min of process in 70% ethanol, then the rice bud is shifted and spike of rice is stripped out in superclean bench, and spike of rice is separated into ramuscule stalk of uniform size, put into after then encasing ramuscule stalk with the gauze that sterilized, with rinsed with sterile water 3～4 times after the completion of sterilizing, open gauze to dry, finally flower pesticide top is cut off with the shears that sterilized, tremble medicine method inoculation flower pesticide in row's Xian culture medium；
（1.3）Inducing culture：Row's Xian culture medium is placed in light culture at 25～27 DEG C, occur the anther callus of yellowish white after 30～40 days at the edge of flower pesticide, and induction of anther callus rate is counted after flower pesticide inoculated and cultured 45 days, the good japonica rice material of economical character of the induction of anther callus rate higher than 35% is selected as japonica rice material to be improved.
The step（1.2）In rice bud refer to that the part of material acquisition cut under for 8cm to fringe by rice bud top pillow in the region of internode top 0.5cm, and the upper flower pesticide quantity of the ramuscule stalk that strips is 15～25.
The step（1.2）In row's Xian culture medium prescriptions be：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0.
The step（2）In Raw toxin patience concentration qualification process as follows：
（2.1）The preparation of Raw toxin：The mixing linum leaf spot fungi that picking is preserved is inoculated on PDA solid mediums, and 25 DEG C of dark culturing are transferred in PDA liquid medium after growing mycelium and producing spore, shaken cultivation 15d under the conditions of 25 DEG C, filters off bacterium with sterile gauze, is centrifuged 20min（1000r/min）, add the activated carbon of about filtrate weight 1/2 in filtrate, 5 DEG C of standing adsorptions 10h, then 8000r/min centrifugations 12min, then by activated carbon precipitation equal-volume methanol-eluted fractions, rotates evaporation and concentration eluent, that is, obtains water rice brown spot bacterium Raw toxin at 60 DEG C；
（2.2）The identification of Raw toxin endurance concentration is carried out to the japonica rice material to be improved that chooses：The Extracted toxin liquid of certain gradient concentration is added in division culture medium respectively, then by step（1）In the anther callus of japonica rice material inducing culture to be improved selected be transferred into the division culture medium, inducing culture counts phenylacetic acid after 30～40 days, and selects the Extracted toxin liquid of suitable screening pressure level concentration to hand over F1 cenospecies Anther Cultures Raw toxin to select pressure as three.
The step（2.2）In Raw toxin select pressure to refer to the selected certain density Extracted toxin liquid for adding division culture medium when the anther callus differentiation rate of japonica rice material inducing culture is improved by 0.7%～1.3%.
The step（2.1）In the compound method of PDA solid mediums be：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, Jia 15～20g agar in filtrate, 20g sucrose adds water and complements to 1000mL；The compound method of described PDA liquid medium is：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, in filtrate plus 20g sucrose, add water and complement to 1000mL；The step（2.2）In differentiation culture based formulas be：The Extracted toxin liquid of the certain gradient concentrations of+0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ of+3% sucrose of MS inorganic salt+MS Organic substances, pH value is 6.0.
The step（4）In three friendship F1 cenospecies induction of anther callus rate be less than 2.5% when, F1 cenospecies need to be handed over to obtain BC1F1 cenospecies with japonica rice parent backcrossing by three, the positive season sowing of BC1F1 cenospecies, choose healthy and strong full flower pesticide and re-start row's Xian culture medium flower pesticide inducing culture, until obtaining the anther callus that inductivity is not less than 2.5%, the anther callus for selecting 0.6～0.9cm of diameter are inoculated in the division culture medium for adding that Raw toxin selects pressure, in 27～29 DEG C of temperature, cultivate under conditions of the dark 10h of light 14h/, obtain flower training seedling, transfer after bloom control height of seedling reaches 3cm successive transfer culture is carried out in root media, transplant to Turnover Box strong sprout after bloom control height of seedling reaches 8cm, land for growing field crops is transplanted into after 10～15 days.
The step（1）With（4）In row's Xian culture medium prescriptions be：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0；The step（4）In differentiation culture based formulas be：+ 0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ Raw toxins of+3% sucrose of MS inorganic salt+MS Organic substances select pressure, and pH value is 6.0；The step（4）In prescription of rooting medium be：1/2Ms micro+0.8% agar of+2% sucrose of+Ms Organic substances of a large amount of+Ms, pH value is 6.0.
Specific embodiment
With reference to embodiment, the present invention is further illustrated.
A kind of Rice Resistance linum leaf spot breeding method, it is characterised in that：The step of breeding method, is as follows：
（1）Select japonica rice material to be improved and the anti-source of water rice brown spot long-grained nonglutinous rice：
Choose that economical character is good, water rice brown spot resistance needs the japonica rice material improved, as Xian round-grained rice hands over F-1 hybrids that generally there is the characteristics of plant height is higher, period of duration is super close late-maturing, thus japonica rice material improve for selecting refer to yield height, good grain quality, linum leaf spot resistance need to be improved and plant height should not too high, period of duration precocious or in ripe japonica rice material；The japonica rice flower pesticide is seeded in row's Xian culture medium（Formula is：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0）Inducing culture, to check whether the japonica rice product material carries out flower pesticide callus induction suitable for row's Xian culture medium, inducing culture process is as follows：
（1.1）Draw materials and pretreatment：The japonica rice material land for growing field crops conventional cultivation that will be chosen, is not broken with rice bud, sword-like leave pulvinus accounts for the 1/3～1/2 of clever shell away from the flower pesticide position as in the middle part of 5～8cm, spike of rice and takes fringe for standard, 2～3 blades of reservation, is taken the fringe time and is typically chosen in the fine day morning 9:00～12:00, after drawing materials, with 70% ethanol surface sterilization, then plastic sheeting encases whole spike of rice, tightens plastic bag mouth, keeps spike of rice moistening, is finally put in refrigerator, Cold pretreatment 3 days at 4 DEG C；
（1.2）Inoculation：After pretreatment, cut material acquisition cut under for 8cm to fringe by top pillow rice bud in the region of internode top 0.5cm, rice bud is soaked in 3～5min of process in 70% ethanol, then the rice bud being shifted, spike of rice is stripped out in superclean bench, and spike of rice is separated into uniform in size, flower pesticide quantity is the ramuscule stalk of 15～25, then put into after ramuscule stalk being encased with the gauze that sterilized, with rinsed with sterile water 3～4 times after the completion of sterilizing, open gauze to dry, finally flower pesticide top is cut off with the shears that sterilized, medicine method inoculation flower pesticide is trembled in row's Xian culture medium.Row's Xian culture medium prescriptions are：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, the pH value for arranging Xian culture medium are 6.0；
（1.3）Inducing culture：Row's Xian culture medium is placed in light culture at 25～27 DEG C, occur the anther callus of yellowish white after 30～40 days at the edge of flower pesticide, and induction of anther callus rate is counted after flower pesticide inoculated and cultured 45 days, the good japonica rice material of economical character of the induction of anther callus rate higher than 35% is selected as japonica rice material to be improved；Japonica rice material of the final choice induction of anther callus rate higher than 35% is used as japonica rice material to be improved, choose that economical character is good and the long-grained nonglutinous rice material of anti-linum leaf spot is used as the anti-source of linum leaf spot long-grained nonglutinous rice, the anti-source of the long-grained nonglutinous rice refers to yield height, good grain quality, the long-grained nonglutinous rice material of high anti-linum leaf spot；
（2）The identification that Raw toxin patience concentration is carried out to the japonica rice material to be improved for selecting：
（2.1）The preparation of Raw toxin：The mixing linum leaf spot fungi that picking is preserved is inoculated on PDA solid mediums（Compound method is：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, Jia 15～20g agar in filtrate, 20g sucrose adds water and complements to 1000mL）, 25 DEG C of dark culturing are transferred in PDA liquid medium after growing mycelium and producing spore（Compound method is：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, in filtrate plus 20g sucrose, add water and complement to 1000mL）, shaken cultivation 15d under the conditions of 25 DEG C, with sterile gauze elimination bacterium, centrifugation 20min（1000r/min）, add the activated carbon of about filtrate weight 1/2 in filtrate, 5 DEG C of standing adsorptions 10h, then 8000r/min centrifugations 12min, then by activated carbon precipitation equal-volume methanol-eluted fractions, rotates evaporation and concentration eluent, that is, obtains water rice brown spot bacterium Raw toxin at 60 DEG C；
（2.2）The identification of Raw toxin endurance concentration is carried out to the japonica rice material to be improved that chooses：The Extracted toxin liquid of certain gradient concentration is added in division culture medium respectively（Formula is：The Extracted toxin liquid of the certain gradient concentrations of+0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ of+3% sucrose of MS inorganic salt+MS Organic substances, pH value is 6.0）, then by step（1）In the anther callus of japonica rice material inducing culture to be improved selected be transferred into the division culture medium, inducing culture counts phenylacetic acid after 30～40 days, and hands over F1 cenospecies Anther Cultures Raw toxin to select pressure as three the certain density Extracted toxin liquid added when the anther callus differentiation rate of japonica rice material inducing culture is improved by 0.7%～1.3% in division culture medium that selectes；
（3）Hybridization obtains three and hands over F1 cenospecies：
Anti- for the linum leaf spot long-grained nonglutinous rice of field planting source is built bridge with wide affinitive material Xian round-grained rice and obtains Hybrid F1, the flower pesticide for choosing Hybrid F1 hands over F1 with japonica rice material to be improved hybridization acquisition cenospecies three again；
（4）The F1 cenospecies row of carrying out Xian culture medium Anther Cultures again are handed over to three：
Three hand over the positive season sowing of F1 cenospecies, choosing healthy and strong full flower pesticide carries out arranging Xian culture medium flower pesticide inducing culture, obtain anther callus, if the induction of anther callus rate of the three friendships F1 cenospecies is less than 2.5%, F1 cenospecies need to be handed over to obtain BC1F1 cenospecies with japonica rice parent backcrossing by three, the positive season sowing of BC1F1 cenospecies, chooses healthy and strong full flower pesticide and re-starts row's Xian culture medium flower pesticide inducing culture（Formula is：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0）, until obtaining the anther callus that inductivity is not less than 2.5%.Then the anther callus for selecting 0.6～0.9cm of diameter are inoculated in the division culture medium that with the addition of selection pressure concentration Raw toxin（Formula is：+ 0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ Raw toxins of+3% sucrose of MS inorganic salt+MS Organic substances select pressure, and pH value is 6.0）On, differentiation culture under conditions of 27～29 DEG C of temperature, the dark 10h of light 14h/ obtains flower training seedling, transfers in root media after bloom control height of seedling reaches 3cm（Formula is：1/2Ms micro+0.8% agar of+2% sucrose of+Ms Organic substances of a large amount of+Ms, pH value is 6.0）Successive transfer culture is carried out, strong sprout in transplanting to Turnover Box after bloom control height of seedling reaches 8cm was transplanted into land for growing field crops after 10～15 days；
（5）Bloom control individual plant sowing obtains flower training family, and further economical character identification, linum leaf spot resistance screening obtain Keng rice lines：
Field planting bloom control, the japonica rice training family that individual plant sowing acquisition linum leaf spot resistance obtains improveing is some, cell plantation in the further land for growing field crops of training family of not suiting, with japonica rice material parent to be improved as control, screen agronomic shape well and linum leaf spot resistance obtains the Keng rice lines of improvement.
The breeding method of the present invention is compared with conventional breeding methods, linum leaf spot genetics of resistance material in long-grained nonglutinous rice efficiently and exclusively can be penetrated into japonica rice, greatly reduce screening operation amount, Rice Resistance linum leaf spot breeding can be completed within 3 years or so, greatly shorten the linum leaf spot breeding for disease resistance cycle, compared with molecular engineering breeding, and have the advantages that operating, threshold is relatively low, it is less to spend, and is worth with larger Breeding Application.
Embodiment
2009～2012, we select rice variety Hunan morning Xian 24 as anti-source, breeding trunk strain H0102 as japonica rice material to be improved respectively, water rice brown spot breeding for disease resistance is carried out using the technology path of the present invention, 2 flower training familys that economical character is excellent, water rice brown spot resistance obtains improvement are have successfully been obtained, its concrete operations flow process is as follows：
（1）H0102 and Hunan morning Xian No. 24 conducts to be improved japonica rice material and water rice brown spot long-grained nonglutinous rice anti-source are screened respectively：
Breeding practice for many years finds, breeding trunk strain H0102 economical character is excellent, yield height, partially meter Zhi Hao, precocity, but compared with sense water rice brown spot, positive season in 2009 by H0102 take flower pesticide be inoculated in row Xian culture medium carry out inducing culture, as a result find H0102 calluss inductivity be 39%（Anther Culture operating procedure is the same）, relatively it is suitable for the requirement of the Japonica two line hybrid rice parent of the technology path.Hunan morning Xian 24 is ripe early Xian new varieties in the high yield that Hunan Rice Research Institute was bred as in 1992, the high water resistant rice brown spot of the rice variety, and have that tillering ability is strong, plant is short, resistant to lodging, blade is straight, period of duration is precocious, stable high yield, protein content are high, it is highly suitable as the requirement in the anti-source of water rice brown spot long-grained nonglutinous rice of this technology route.
（2）The identification that Raw toxin patience concentration is carried out to H0102：
Positive season in 2009 carries out the identification of Raw toxin endurance concentration to the H0102 for choosing, and adds 5%, 10%, 15%, 20%, 25% Extracted toxin liquid, prepare division culture medium in division culture medium, and differentiation culture based formulas are：The certain gradient concentration Extracted toxin liquid of+0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ of+3% sucrose of MS inorganic salt+MS Organic substances, pH value 6.0, then by step（1）The anther callus of middle inducing culture are transferred into division culture medium, count phenylacetic acid.As a result such as following table：
Further south is numerous for H0102, and obtaining flower pesticide carries out inducing culture, and the further optimization of Raw toxin patience concentration, as a result as follows：
Therefore, 17% Extracted toxin liquid concentration is set to three friendship F1 cenospecies Anther Cultures Raw toxins and selects pressure by Integrated comparative, we.
（3）Cross-breeding obtains three and hands over F1 cenospecies：
Hunan morning Xian in 2009 24 is built bridge with extensively affine japonica rice 02428 of short stem hybridization, harvests Hybrid F1, and numerous the selections F1 flower pesticide of South of Hainan in 2010 obtains the friendship F1 of cenospecies three again with H0102 hybridization.
（4）Three friendship F1 cenospecies are carried out arranging Xian culture medium Anther Cultures：
Positive season land for growing field crops general planting three in 2010 hands over F1 cenospecies, and the flower pesticide of the healthy and strong full tassel of selection is inoculated in row's Xian culture medium carries out flower pesticide inducing culture acquisition anther callus, and row's Xian culture medium prescriptions are：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0；And select the anther callus of 0.6～0.9cm of diameter and be inoculated in the division culture medium that with the addition of 17% Extracted toxin liquid, differentiation culture based formulas are：The Extracted toxin liquid of+0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+17% of+3% sucrose of MS inorganic salt+MS Organic substances, pH value is 6.0, continue differentiation culture under conditions of 27～29 DEG C of temperature, the dark 10h of light 14h/, obtain 123 flower training seedling；Transfer after bloom control height of seedling reaches 3cm successive transfer culture is carried out in root media, 1/2Ms micro+0.8% agar of+2% sucrose of+Ms Organic substances of a large amount of+Ms, pH value is 6.0, transplants to Turnover Box strong sprout, be transplanted into greenhouse within 2011 years after bloom control length to 8cm.
（5）Flower training family field planting, economical character, linum leaf spot resistance screening obtain 2 excellent Keng rice lines of linum leaf spot resistance improvement：
It is transplanted to the bloom control individual plant sowing in greenhouse, obtain 114 flower training familys, positive season in 2012 114 flower training familys of further field seeding, with H0102 as control, therefrom screening obtains that 2 agronomic shapes are excellent and water rice brown spot resistance obtains the Keng rice lines improved.
From the present embodiment, the breeding method of the present invention is compared with conventional breeding methods, linum leaf spot genetics of resistance material in long-grained nonglutinous rice efficiently and exclusively can be penetrated into japonica rice, greatly reduce screening operation amount, only Rice Resistance linum leaf spot breeding can be completed with 3 years, so as to greatly shorten the linum leaf spot breeding for disease resistance cycle, and operation threshold is relatively low, it is less to spend, so being worth with larger Breeding Application.
Above example technological thought only to illustrate the invention, it is impossible to which protection scope of the present invention is limited with this, every according to technological thought proposed by the present invention, any change that is done on the basis of technical scheme, each fall within the scope of the present invention；The technology that the present invention is not directed to can be realized by prior art.
Claims (10)
1. a kind of Rice Resistance linum leaf spot breeding method, it is characterised in that：The step of breeding method, is as follows：
（1）Select japonica rice material to be improved and the anti-source of water rice brown spot long-grained nonglutinous rice：Choose that economical character is good, water rice brown spot resistance needs the japonica rice material improved and the japonica rice flower pesticide is seeded to inducing culture in row's Xian culture medium, used as japonica rice material improve, selection economical character is good and the long-grained nonglutinous rice material of anti-linum leaf spot is used as the anti-source of linum leaf spot long-grained nonglutinous rice for japonica rice material of the final choice induction of anther callus rate higher than 35%；
（2）The identification that Raw toxin patience concentration is carried out to the japonica rice material to be improved for selecting：The Extracted toxin liquid of certain gradient concentration is added in division culture medium respectively, to step（1）In the anther callus that obtain of japonica rice material to be improved induction carry out differentiation culture, and select the Extracted toxin liquid of suitable screening pressure level concentration to select pressure as japonica rice Anther Culture Raw toxin；
（3）Hybridization obtains three and hands over F1 cenospecies：Anti- for the linum leaf spot long-grained nonglutinous rice of field planting source is built bridge with wide affinitive material Xian round-grained rice and obtains Hybrid F1, the flower pesticide for choosing Hybrid F1 hands over F1 with japonica rice material to be improved hybridization acquisition cenospecies three again；
（4）The F1 cenospecies row of carrying out Xian culture medium Anther Cultures again are handed over to three：Three hand over the positive season sowing of F1 cenospecies, choosing healthy and strong full flower pesticide carries out arranging Xian culture medium flower pesticide inducing culture, obtain anther callus, and select the anther callus of 0.6～0.9cm of diameter be inoculated in the addition of select pressure concentration Raw toxin division culture medium on, differentiation culture under conditions of 27～29 DEG C of temperature, the dark 10h of light 14h/, obtain flower training seedling, transfer after bloom control height of seedling reaches 3cm successive transfer culture is carried out in root media, strong sprout in transplanting to Turnover Box after bloom control height of seedling reaches 8cm, was transplanted into land for growing field crops after 10～15 days；
（5）Bloom control individual plant sowing obtains flower training family, and further economical character identification, linum leaf spot resistance screening obtain Keng rice lines：Field planting bloom control, the japonica rice training family that individual plant sowing acquisition linum leaf spot resistance obtains improveing is some, cell plantation in the further land for growing field crops of training family of not suiting, with japonica rice material parent to be improved as control, screen agronomic shape well and linum leaf spot resistance obtains the Keng rice lines of improvement.
2. Rice Resistance linum leaf spot breeding method according to claim 1, it is characterised in that：The step（1）In economical character is good, the japonica rice material that water rice brown spot resistance needs to be improved refer to yield height, good grain quality, linum leaf spot resistance need to be improved and plant height should not too high, period of duration precocious or in ripe japonica rice material；The anti-source of described water rice brown spot long-grained nonglutinous rice refers to yield height, good grain quality, the long-grained nonglutinous rice material of high anti-linum leaf spot.
3. Rice Resistance linum leaf spot breeding method according to claim 1, it is characterised in that：The step（1）The process that middle selection economical character is good, water rice brown spot resistance needs the japonica rice material that is improved and the japonica rice flower pesticide is seeded to inducing culture in row's Xian culture medium is as follows：
（1.1）Draw materials and pretreatment：The japonica rice material land for growing field crops conventional cultivation that will be chosen, is not broken with rice bud, sword-like leave pulvinus accounts for the 1/3～1/2 of clever shell away from the flower pesticide position as in the middle part of 5～8cm, spike of rice and takes fringe for standard, 2～3 blades of reservation, is taken the fringe time and is typically chosen in the fine day morning 9:00～12:00, after drawing materials, with 70% ethanol surface sterilization, then plastic sheeting encases whole spike of rice, tightens plastic bag mouth, keeps spike of rice moistening, is finally put in refrigerator, Cold pretreatment 3 days at 4 DEG C；
（1.2）Inoculation：After pretreatment, cut rice bud, rice bud is soaked in 3～5min of process in 70% ethanol, then the rice bud is shifted and spike of rice is stripped out in superclean bench, and spike of rice is separated into ramuscule stalk of uniform size, put into after then encasing ramuscule stalk with the gauze that sterilized, with rinsed with sterile water 3～4 times after the completion of sterilizing, open gauze to dry, finally flower pesticide top is cut off with the shears that sterilized, tremble medicine method inoculation flower pesticide in row's Xian culture medium；
（1.3）Inducing culture：Row's Xian culture medium is placed in light culture at 25～27 DEG C, occur the anther callus of yellowish white after 30～40 days at the edge of flower pesticide, and induction of anther callus rate is counted after flower pesticide inoculated and cultured 45 days, the good japonica rice material of economical character of the induction of anther callus rate higher than 35% is selected as japonica rice material to be improved.
4. Rice Resistance linum leaf spot breeding method according to claim 3, it is characterised in that：The step（1.2）In rice bud refer to that the part of material acquisition cut under for 8cm to fringe by rice bud top pillow in the region of internode top 0.5cm, and the upper flower pesticide quantity of the ramuscule stalk that strips is 15～25.
5. Rice Resistance linum leaf spot breeding method according to claim 3, it is characterised in that：The step（1.2）In row's Xian culture medium prescriptions be：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0.
6. Rice Resistance linum leaf spot breeding method according to claim 1, it is characterised in that：The step（2）In Raw toxin patience concentration qualification process as follows：
（2.1）The preparation of Raw toxin：The mixing linum leaf spot fungi that picking is preserved is inoculated on PDA solid mediums, and 25 DEG C of dark culturing are transferred in PDA liquid medium after growing mycelium and producing spore, shaken cultivation 15d under the conditions of 25 DEG C, filters off bacterium with sterile gauze, is centrifuged 20min（1000r/min）, add the activated carbon of about filtrate weight 1/2 in filtrate, 5 DEG C of standing adsorptions 10h, then 8000r/min centrifugations 12min, then by activated carbon precipitation equal-volume methanol-eluted fractions, rotates evaporation and concentration eluent, that is, obtains water rice brown spot bacterium Raw toxin at 60 DEG C；
（2.2）The identification of Raw toxin endurance concentration is carried out to the japonica rice material to be improved that chooses：The Extracted toxin liquid of certain gradient concentration is added in division culture medium respectively, then by step（1）In the anther callus of japonica rice material inducing culture to be improved selected be transferred into the division culture medium, inducing culture counts phenylacetic acid after 30～40 days, and selects the Extracted toxin liquid of suitable screening pressure level concentration to hand over F1 cenospecies Anther Cultures Raw toxin to select pressure as three.
7. Rice Resistance linum leaf spot breeding method according to claim 6, it is characterised in that：The step（2.2）In Raw toxin select pressure to refer to the selected certain density Extracted toxin liquid for adding division culture medium when the anther callus differentiation rate of japonica rice material inducing culture is improved by 0.7%～1.3%.
8. Rice Resistance linum leaf spot breeding method according to claim 6, it is characterised in that：The step（2.1）In the compound method of PDA solid mediums be：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, Jia 15～20g agar in filtrate, 20g sucrose adds water and complements to 1000mL；The compound method of described PDA liquid medium is：Rhizoma Solani tuber osi is cleaned peeling to be cut into small pieces, is weighed 200g, is put in pot, add water 1000mL, boils 30min, slightly cold after filtered with double gauze, in filtrate plus 20g sucrose, add water and complement to 1000mL；The step（2.2）In differentiation culture based formulas be：The Extracted toxin liquid of the certain gradient concentrations of+0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ of+3% sucrose of MS inorganic salt+MS Organic substances, pH value is 6.0.
9. Rice Resistance linum leaf spot breeding method according to claim 1, it is characterised in that：The step（4）In three friendship F1 cenospecies induction of anther callus rate be less than 2.5% when, F1 cenospecies need to be handed over to obtain BC1F1 cenospecies with japonica rice parent backcrossing by three, the positive season sowing of BC1F1 cenospecies, choose healthy and strong full flower pesticide and re-start row's Xian culture medium flower pesticide inducing culture, until obtaining the anther callus that inductivity is not less than 2.5%, the anther callus for selecting 0.6～0.9cm of diameter are inoculated in the division culture medium for adding that Raw toxin selects pressure, in 27～29 DEG C of temperature, cultivate under conditions of the dark 10h of light 14h/, obtain flower training seedling, transfer after bloom control height of seedling reaches 3cm successive transfer culture is carried out in root media, transplant to Turnover Box strong sprout after bloom control height of seedling reaches 8cm, land for growing field crops is transplanted into after 10～15 days.
10. Rice Resistance linum leaf spot breeding method according to claim 1, it is characterised in that：The step（1）With（4）In row's Xian culture medium prescriptions be：N6 a great number of elements+MS trace element+N6 organic matter+1.8mg/L2 ,+0.7% agar of+5% sucrose of 4-D+0.4mg/LNAA+0.3mg/L hydrolyzed proteins, pH value is 6.0；The step（4）In differentiation culture based formulas be：+ 0.6% plant gel+KT2.0mg/L+NAA0.5mg/L+ hydrolyzed protein 1g/L+ Raw toxins of+3% sucrose of MS inorganic salt+MS Organic substances select pressure, and pH value is 6.0；The step（4）In prescription of rooting medium be：1/2Ms micro+0.8% agar of+2% sucrose of+Ms Organic substances of a large amount of+Ms, pH value is 6.0.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107148908A (en) * | 2017-06-14 | 2017-09-12 | 江苏强农农业技术服务有限公司 | A kind of scab resistance wheat breeding new method |
| CN107182774A (en) * | 2017-06-14 | 2017-09-22 | 江苏强农农业技术服务有限公司 | The mould leaf blight resistant wheat breeding new method of one kind snow |
| CN107211882A (en) * | 2017-06-14 | 2017-09-29 | 尹纪岩 | A kind of utilization pine root fungus Raw toxin quickly screens the breeding method of root rotof flax resistant variety |
| CN110033825A (en) * | 2019-04-26 | 2019-07-19 | 扬州大学 | A kind of Jian Kuo pea Screening of Germplasm method |
| CN112806259A (en) * | 2021-01-20 | 2021-05-18 | 云南帅豪农牧科技有限公司 | High-disease-resistance polygonatum sibiricum tissue culture seedling method |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101250535A (en) * | 2008-04-07 | 2008-08-27 | 华中农业大学 | Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance |
| CN104396970A (en) * | 2014-11-11 | 2015-03-11 | 浙江新农化工股份有限公司 | Sterilizing composition containing zinc thiazole, preparation and applications thereof |
| CN104542256A (en) * | 2015-01-04 | 2015-04-29 | 陈丁龙 | Breeding method for importing false smut resistance of indica rice into japonica rice |
-
2015
- 2015-09-08 CN CN201510562533.3A patent/CN106489716A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101250535A (en) * | 2008-04-07 | 2008-08-27 | 华中农业大学 | Rice disease-resistant related gene OsDR9 and its use in improving rice disease resistance |
| CN104396970A (en) * | 2014-11-11 | 2015-03-11 | 浙江新农化工股份有限公司 | Sterilizing composition containing zinc thiazole, preparation and applications thereof |
| CN104542256A (en) * | 2015-01-04 | 2015-04-29 | 陈丁龙 | Breeding method for importing false smut resistance of indica rice into japonica rice |
Non-Patent Citations (2)
| Title |
|---|
| 凌定厚: "运用植物毒素离体筛选水稻抗虎妈叶斑病种质的研究", 《遗传学报》 * |
| 赵晓宇等: "水稻胡麻叶斑病的发生及防治措施", 《北方水稻》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107148908A (en) * | 2017-06-14 | 2017-09-12 | 江苏强农农业技术服务有限公司 | A kind of scab resistance wheat breeding new method |
| CN107182774A (en) * | 2017-06-14 | 2017-09-22 | 江苏强农农业技术服务有限公司 | The mould leaf blight resistant wheat breeding new method of one kind snow |
| CN107211882A (en) * | 2017-06-14 | 2017-09-29 | 尹纪岩 | A kind of utilization pine root fungus Raw toxin quickly screens the breeding method of root rotof flax resistant variety |
| CN110033825A (en) * | 2019-04-26 | 2019-07-19 | 扬州大学 | A kind of Jian Kuo pea Screening of Germplasm method |
| CN110033825B (en) * | 2019-04-26 | 2021-08-13 | 扬州大学 | Method for screening germplasm resources of common vetch |
| CN112806259A (en) * | 2021-01-20 | 2021-05-18 | 云南帅豪农牧科技有限公司 | High-disease-resistance polygonatum sibiricum tissue culture seedling method |
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