CN104170668A - Efficient smut inoculation method for sugarcane seedlings - Google Patents
Efficient smut inoculation method for sugarcane seedlings Download PDFInfo
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- CN104170668A CN104170668A CN201410365980.5A CN201410365980A CN104170668A CN 104170668 A CN104170668 A CN 104170668A CN 201410365980 A CN201410365980 A CN 201410365980A CN 104170668 A CN104170668 A CN 104170668A
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Abstract
A smut inoculation method for sugarcane seedlings comprises the steps as follows: after seven true leaves of each seedling are grown out, the seedling is pulled out from an incubator, the middle-upper parts of the leaves are cut off, 2-3 old leaves at the base part are removed, a spore suspension with the ustilago scitaminea syd concentration of 1*10<7> spores/ml is adopted for soaking the middle-lower part of the pulled seedling for 10 min, and the seedling is sealed, placed in a room at the temperature of 25-28 DEG C overnight in a moisture preserving manner, transplanted in a nutrient bag next day and planted in the field after survived; and combination morbidity is researched during final selection, combination with the combination morbidity higher than 25% is eliminated, and seedlings with good and healthy characters are screened out to enter a next stage for observation. According to the invention, morbidity of the seedlings inoculated with ustilago scitaminea syd are remarkable, the reproducibility is good, the test cycle is short, the operability is high, the cost is low, and a large quantity of seedlings can be inoculated; and the method can be applied to a sugarcane variety breeding institution for new sugarcane variety breeding resistant to sugarcane smut.
Description
Technical field:
The present invention relates to smut resistance in sugarcane breed breeding field, is that a kind of cane seedlings carries out ustilago scitaminea bacteria high-efficiency inoculating method, reaches the method for early stage selection smut resistance combination and individual plant.
Technical background:
Culmicolous smut of sugar cane is a kind of gas borne fungus diseases being caused by smut (Ustilago scitaminea Syd.), this disease becomes the topmost fungal disease in all countries and regions that grow cane, eliminated once the govern-house-variety of the high sugar of some high yields, cause the heavy losses of susceptible variety cane yield and Sucrose, on upland sugarcane, cause harm even more serious, generally acknowledge that controlling the most economical effective approach of this disease is plantation disease-resistant variety.The U.S., Brazil, the main Chan Zhe state such as Cuba and China is one of main target using smut resistance as cane breeding all.Seed selection smut resistance sugar cane breed, to the healthy stable development of promotion industry, ensures that sugar safety is significant.(1) smut resistance in sugarcane breeding approach has by conventional breeding approach, the disease-resistant good sugarcane germplasm material filtering out is carried out to sexual hybridization, and its offspring is carried out to Disease Resistance Identification.After the improved seeds (being) that obtain after the screening of " five gardens " system are accredited as disease-resistant variety, apply, required time is longer; (2) by animal nutrition, obtain disease-resistant gene, disease-resistant foreign gene is imported in Sugarcane smut, but also need to solve the foreign gene proceeding to, whether reach the problem that expectation function is expressed.So far. on smut resistance in sugarcane breeding practice, adopt in generation morning of breeding, utilize natural infection method to eliminate susceptible strain; In the height generation of breeding, adopt artificial infection to carry out the method for field resistance approval test, further eliminate susceptible material.Its deficiency is that breeding cycle is long, and seed selection smut resistance kind needs 8 years above time; And the smut resistance selection pressure applying due to early stage is little, the later stage adopts artificial infection field resistance to identify, a lot of material sense smut, has wasted a large amount of manpower and materials, and Breeding Efficiency is low.In order to improve smut resistance sugar cane breed Breeding Efficiency, Sugarcane research institute has carried out the research work of seedling early screening smut resistance material, formed and before cane seedlings field-transplanting, carried out culmicolous smut of sugar cane inoculation and coerce method, behind field planting land for growing field crops according to incidence, eliminate poor combination and the individual plant of disease resistance, make smut resistance screening critical point in advance, improve Breeding Efficiency, shortening the breeding cycle.
Sugarcane Inst., Yunnan Prov. Agriculture Academy is more than 800 for the sugarcane hybrid combination of breed breeding in recent years every year, and training the region between the heart and the diaphragm seedling quantity is 13-15 ten thousand seedlings, and more than 50 mu of left and right is taken up an area in optional seedlings field planting.Because hybrid combination amount is large, with traditional authentication method, need to expend substantial contribution and labour and drop into.
Summary of the invention:
The object of the invention is for above-mentioned the deficiencies in the prior art, for the early stage breeding for disease resistance of smut resistance in sugarcane individual plant and combination of selecting provides a kind of efficient, easy method that cane seedlings is carried out to smut inoculation.
The present invention adopts following technical scheme to be achieved: after seedling growth to 7 true leaf, the tiller bud of seedling is carried out to smut inoculation, make smut germ invade the tiller bud of dormancy, transplant next day in nutritious bag, survive rear field planting land for growing field crops, when final election, filter out seedling combination and the individual plant of smut resistance, from incubator, extract seedling and cut off the middle and upper part of leaf, water washes out after the soil of root, remove base portion Lao Ye 2-3 leaf, the tiller bud of base portion dormancy is exposed, the spore suspension that is 1 * 107 spore/ml by black tassel bacteria concentration soaks 10 minutes the seedling middle and lower part of extracting, and pack in plastic sack and seal, the indoor moisturizing that is placed on 25-28 ℃ is spent the night, make smut germ invade the tiller bud of dormancy, the investigation combination incidence of disease when final election, the combination incidence of disease is greater than to 25% combination to be eliminated and not to select, again in conjunction with other economical character, the seedling strain that filters out the good health of proterties enters next stage and observes, thereby Preliminary screening goes out seedling combination and the individual plant of smut resistance.Reference when seedling combines susceptible situation as the disease-resistant combination of later preparation sugarcane.
The present invention has following beneficial effect:
There is the tiller bud of a plurality of dormancy in (1) 7 leaf phase seedling base portion, by immersion, inoculate black tassel bacteria spore, behind field planting land for growing field crops, smut germ is invaded the tiller bud of sprouting, increase the chance that seedling infects culmicolous smut of sugar cane, the time that in test, smut whip produces wants Zao than the inoculation of contrast stem eye, and occurring degree is heavier, reach the effect that inoculation black tassel bacteria is coerced cane seedlings;
(2) seedling is soaked to inoculation black tassel bacteria spore, easy to operate, cost is low, can inoculate a large amount of seedlings, the smut spore suspension of 5 liters just can be inoculated the seedling of seedling more than 10,000, can reduce the cost burden of a large amount of collection smut spores;
(3) commitment is by eliminating susceptible seedling combination and individual plant, thereby Preliminary screening goes out seedling combination and the individual plant of smut resistance, shorten the breeding time limit of seed selection smut resistance new variety of sugarcane, improve breeding efficiency, effectively save seed selection new variety of sugarcane practice ground and later stage fund input;
(4) seedling inoculation black tassel bacteria sequela is remarkable, and repeatability is good, the test period is short, workable;
The method can be served sugar cane breed seed selection mechanism, for the New Sugarcane Breeding of smut resistance in sugarcane.
Embodiment:
1, for examination cane seedlings material, prepare
According to breeding objective, carry out Sugarcane Breeding combination selection, after carry out sexual hybridization and obtain hybrid seed.Hybrid seed is seeded in the container that cane seedlings culture matrix is housed.Control and cultivate indoor temperature 22-36 ℃, and keep matrix moist, carry out the extermination of disease and insect pest in good time.Can obtain cane seedlings.
2, prepare smut germ spore suspension
(1) gather culmicolous smut of sugar cane germ spore
Collect smut germ spore, dry Conidia preservation, in being placed with the bottle of silica-gel desiccant, is sealed with vaseline.
(2) before inoculation, smut germ spore suspension is prepared
Before test, spore is carried out to survival test.And to prepare smut germ concentration be 1 * 10
7the spore suspension of spore/ml.
3, method is coerced in inoculation
After seedling growth to 7 true leaf, from incubator, extract the middle and upper part that seedling cuts off leaf, water washes out after the soil of root, removes base portion Lao Ye 2-3 leaf, and the tiller bud of base portion dormancy is exposed, and by black tassel bacteria concentration, is 1 * 10
7the spore suspension of spore/ml soaks 10 minutes the seedling middle and lower part of extracting, and pack in plastic sack and seal, the indoor moisturizing that is placed on 25-28 ℃ is spent the night, make smut germ invade the tiller bud of dormancy, transplant next day in nutritious bag, initial stage shelters from heat or light with shade net, survives rear field planting land for growing field crops, reaches the effect that inoculation smut germ is coerced cane seedlings.
4, the system of selection of the combination of the seedling of smut resistance and individual plant
The sick rate of investigation combination burst when final election, the combination incidence of disease is greater than to 25% combination to be eliminated and not to select, again in conjunction with other economical character, the seedling strain that filters out the good health of proterties enters next stage and observes, thereby Preliminary screening goes out seedling combination and the individual plant of smut resistance.Reference when seedling combines susceptible situation as the disease-resistant combination of later preparation sugarcane.
Research shows, smut resistance in sugarcane proterties belongs to the proterties that heritability is higher, is suitable for sexual generation to select.Its test selects each 4 of disease-resistant and susceptible variety to hybridize, and obtains the susceptible percentage of offspring's smut as follows: disease-resistant * disease-resistant is 13.8%; Disease-resistant * susceptible is 27.8%; Susceptible * disease-resistant is 41.4%; Susceptible * susceptible is 80.2%.Visible, if two parents are disease-resistant, the susceptible percentage of offspring's smut is minimum, and its hybrid generation's resistance against diseases is strong; Two parents are susceptible, and the susceptible percentage of offspring's smut is the highest, and its hybrid generation's resistance against diseases is poor; One of two parents are disease-resistant, and the susceptible percentage of offspring's smut is medium.In the practices of breeding, can eliminate the poor combination of resistance in F1 colony, and by eliminating susceptible strain, can improve largely the disease-resistant level of the latter half breeding population.
Resistance stability study shows, the extreme resistance level of tool is as high in its recurrence rate of material of high resistance or high sense.Secondly be anti-or sense, the recurrence rate of medium resistance type is extremely low.The material that the extreme resistance of tool is described just can obtain more believable result by the inoculated identification of less number of times, and seedling early screening smut resistance strain has rationale.
Before cane seedlings is transplanted land for growing field crops, carrying out black tassel bacteria inoculation coerces, the combination that resistance is poor, filial generation morbidity is many, according to the susceptible performance of plant, in the first stage of Sugarcane Breeding program, just eliminate susceptible combination and individual plant, thereby Preliminary screening goes out seedling combination and the individual plant of smut resistance, shorten smut resistance kind and the material breeding time limit, improve breeding for disease resistance efficiency of selection.
There is the tiller bud of a plurality of dormancy in 7 leaf phase seedling base portions, by immersion, inoculates black tassel bacteria spore, and behind field planting land for growing field crops, sprouting of tiller bud provides possibility for spore infects, and the time that in test, smut whip produces wants Zao than stem eye inoculation.
The inoculation method of innovating by employing, through 3 years, the test of a plurality of combinations, filter out the germ concentration that effect of inoculation is good, the technical key points such as the combination incidence of disease that inoculation time and superseded disease resistance are poor, formed efficient, repeatability is good, workable, the method of the seedling early screening smut resistance material that cost is low, 22 combinations of participating in the experiment in test, the average clump incidence of disease 41.7%, the highest combination clump incidence of disease is 81.8%, inoculation black tassel bacteria sequela is remarkable, inoculate easy and simple to handle, by eliminating poor combination and the individual plant of resistance, saved soil, fund and labour, improve smut resistance sugar cane breed Breeding Efficiency.
The applicant " carries out culmicolous smut of sugar cane inoculation and coerces method " (the patent No.: problem ZL201110191326.3) existing has in practice been done improvement for the patent of application in 2011 before cane seedlings field-transplanting, the present invention compares with this patent, from aspects such as vaccination ways, infection process temperature and morbidity effects, more all there is outstanding substantive distinguishing features and progressive significantly, table specific as follows:
Claims (1)
1. a method of cane seedlings being carried out to smut inoculation, it is characterized in that, after seedling growth to 7 true leaf, the tiller bud of seedling is carried out to smut inoculation, make smut germ invade the tiller bud of dormancy, transplant next day in nutritious bag, survive rear field planting land for growing field crops, when final election, filter out seedling combination and the individual plant of smut resistance, from incubator, extract seedling and cut off the middle and upper part of leaf, water washes out after the soil of root, removes base portion Lao Ye 2-3 leaf, the tiller bud of base portion dormancy is exposed, and is 1 * 10 by black tassel bacteria concentration
7the spore suspension of spore/ml soaks 10 minutes the seedling middle and lower part of extracting, and pack in plastic sack and seal, the indoor moisturizing that is placed on 25-28 ℃ is spent the night, make smut germ invade the tiller bud of dormancy, the investigation combination incidence of disease when final election, the combination incidence of disease is greater than to 25% combination and eliminates and do not select, filter out the seedling strain of the good health of proterties.
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104838909A (en) * | 2015-05-22 | 2015-08-19 | 云南省农业科学院甘蔗研究所 | Brown streak inoculation stress method before field transplanting of sugarcane seedlings |
| CN105613072A (en) * | 2016-01-20 | 2016-06-01 | 安徽理想种业有限公司 | Identification method of resistance to rice blast |
| CN106544314A (en) * | 2016-10-20 | 2017-03-29 | 华南农业大学 | A kind of ustilago scitaminea bacteria teleutospore collects storage practice |
| CN109355350A (en) * | 2018-11-08 | 2019-02-19 | 云南省农业科学院甘蔗研究所 | A kind of smut resistance in sugarcane rapid identification method |
| CN113142002A (en) * | 2021-01-26 | 2021-07-23 | 广西壮族自治区农业科学院 | Sugarcane smut resistance determination method based on Internet of things system |
| CN114847115A (en) * | 2021-02-03 | 2022-08-05 | 广西大学 | High-flux quick and simple inoculation method for sugarcane smut bacteria and application of high-flux quick and simple inoculation method |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104838909A (en) * | 2015-05-22 | 2015-08-19 | 云南省农业科学院甘蔗研究所 | Brown streak inoculation stress method before field transplanting of sugarcane seedlings |
| CN105613072A (en) * | 2016-01-20 | 2016-06-01 | 安徽理想种业有限公司 | Identification method of resistance to rice blast |
| CN106544314A (en) * | 2016-10-20 | 2017-03-29 | 华南农业大学 | A kind of ustilago scitaminea bacteria teleutospore collects storage practice |
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| CN113142002A (en) * | 2021-01-26 | 2021-07-23 | 广西壮族自治区农业科学院 | Sugarcane smut resistance determination method based on Internet of things system |
| CN114847115A (en) * | 2021-02-03 | 2022-08-05 | 广西大学 | High-flux quick and simple inoculation method for sugarcane smut bacteria and application of high-flux quick and simple inoculation method |
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Application publication date: 20141203 |