CN103718848A - Alfalfa breeding method using cold plasma processing - Google Patents
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Abstract
The invention discloses an alfalfa breeding method using cold plasma processing. The alfalfa breeding method using the cold plasma processing disclosed by the invention comprises the following steps: (1) carrying out the cold plasma processing on alfalfa raw materials to be processed to obtain the materials subjected to the cold plasma processing; and (2) selecting alfalfas with the target property from the materials subjected to the cold plasma processing. The alfalfa breeding method using the cold plasma processing disclosed by the invention breaks through the potential dangers that the traditional breeding has a low level and is repeated, the homogenization is serious and the transgenosis breeding is carried out, and overcomes the disadvantage that the economic input of space breeding is great; the breeding period can be shortened; the alfalfa breeding method using the cold plasma processing has the advancement in the aspect of breeding methods and means.
Description
Technical field
The present invention relates to a kind of Alfalfa Breeding method that cold plasma is processed.
Background technology
Traditional breeding method has very large achievement in the past few decades really, but due to limitation such as breeding time are long, efficiency is lower, only by rule of thumb, relatively blindly, causes low-level repetition, homogeneity serious, and traditional breeding method runs into very large bottleneck at present, even stagnates.Want has higher achievement aspect breeding, just must rely on modern science and technology.But, there is at present breakthrough transgenic breeding method and caused in the world heated argument, think that genetically modified crops have great potential danger, may threaten to human health and environment for human survival.Development along with space technology, fine-variety breeding is carried out in the mutagenesis that utilizes the special environmental condition of space to produce plant seed becomes a kind of emerging breeding method, yet space breeding is a kind of new method of induced mutations breeding, the generation of all induced mutationss is all random, also do not accomplish at present directed mutagenesis, the success rate of space breeding is along with the difference of species also has very big difference, and the cost of space breeding is quite high, and not general entity and individual can bear.
In this case, how to find a kind of traditional biological breeding low-level repetition operation of can breaking through, do not change again the breeding technique of gene character, safety and environmental protection, become the task of top priority.
Summary of the invention
The object of this invention is to provide a kind of Alfalfa Breeding method that cold plasma is processed.
The Alfalfa Breeding method that a kind of cold plasma provided by the invention is processed, comprises the steps:
(1) pending clover raw material are carried out to cold plasma processing, obtain the material after cold plasma is processed;
(2) in the material from cold plasma is processed, seed selection has the clover of objective trait.
In said method, the condition that described cold plasma is processed is: take helium as working media, in vacuum sealing environment, the unionized width that under the processing power of 1-500W, clover raw material is carried out to 15-20s is penetrated processing.
In above-mentioned arbitrary described method, described objective trait for as lower at least one: with without the comparing with a collection of clover raw material of any processing, alfalfa seed germinating capacity improves, clover plant height improves, alfalfa output improves and clover hybrid seed yield improves;
Described processing power is 10-400W, is specially 10-300W.
In above-mentioned arbitrary described method, described processing power is 10-400W, processing time is while being 15-20s, with without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves and/or the plant height of clover improves and/or the output of clover improves and/or the hybrid seed yield of clover increases.
In above-mentioned arbitrary described method, described processing power is 10-300W, processing time is while being 15-20s, with without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves and/or the plant height of clover improves and/or the output of clover improves and/or the hybrid seed yield of clover increases.
In above-mentioned arbitrary described method, described processing power is 10-200W, and the processing time is while being 15-18s, and without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
In above-mentioned arbitrary described method, described processing power is 10-160W, processing time is 15-18s, or processing power is 100-300W, and the processing time is 18-20s, or processing power is 100-200W, processing time is 18s, or processing power is 100-160W, and the processing time is while being 18s, with without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
In above-mentioned arbitrary described method, described processing power is 20W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves;
Described processing power is 80W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the plant height of the clover after processing improves;
Described processing power is 20W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the output of the clover after processing improves;
Described reason power is respectively 100W, 160W or 200W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
The method that in material described in above-mentioned steps (2) from cold plasma is processed, seed selection has a clover of objective trait can be prior art, as to as described in the material of cold plasma after processing carry out adding generation, and utilize DNA molecular marker technology and Identification of Isozyme seed selection to there is the stable inbred line of objective trait; The stable inbred line with objective trait is carried out to selfing purification and rejuvenation again, described in finally obtaining, there is the clover of objective trait.
In above-mentioned arbitrary described method, described alfalfa seed germinating capacity is alfalfa seed germination vigor and/or germination rate;
Described alfalfa output is presented as hay rate of growth and/or fresh careless rate of growth;
Described clover hybrid seed yield is presented as the number that bears pods;
The seed that described clover raw material are clover.
The application of above-mentioned arbitrary described method in Alfalfa Breeding also belongs to protection scope of the present invention.
The advantage of the Alfalfa Breeding method that a kind of cold plasma provided by the invention is processed is as follows:
1, cold plasma processing power is artificially controlled, by repetition test, can find the processing power of a certain specific trait of certain alfalfa seed, can meet better the demand of seed selection clover improved seeds, overcome traditional breeding technique time length, target blindness, breeding quality is low, breeding cost is high drawback, on breeding technique, there is novelty.
2, utilize cold plasma technology to process inbred line seed and can excite seed vitality, realize short long, degeneration-resistant effect, the alfalfa seed germination vigor being bred as, germination rate increase, resistance strengthens, output increases, quality improving, has filled up the blank that in field of agricultural sciences, the unionized width technology of penetrating is applied in breeding.
3, pollution-free, low, the low-carbon (LC) that consumes energy, genomic dna sequence does not change, and the change occurring by gene expression is brought out clover proterties in the present age and is changed, and has overcome the potential danger of genetically modified crops, can not threaten to human health and environment for human survival.
4, this method, again by auxiliary additive selections such as molecular labeling and Identification of Isozymes, further shortens breeding cycle, accelerates breeding speed, and can select more good New alfalfa cultivars.
Method provided by the invention has broken through traditional breeding method low-level repetition, homogeneity is serious and transgenic breeding potential danger, and the huge shortcoming of space breeding economic input, can shorten breeding cycle, in breeding method and means, has advance.
Embodiment
The experimental technique using in following embodiment if no special instructions, is conventional method.
In following embodiment, material used, reagent etc., if no special instructions, all can obtain from commercial channels.
No. 1 alfalfa of middle lucerne was passed through authorization in Ningxia in 2003, authorization (registration) numbering: rather examine lucerne 2003006, the acquisition of public Ke Cong China Agricultural University.
No. 2 alfalfas of middle lucerne are the authorization through national herbage variety validation board in December, 2003, is registered as improved variety, and public Ke Cong China Agricultural University obtains.
No. 3, middle lucerne alfalfa record date is on December 13rd, 2006, and cultivar registration number is 321, and public Ke Cong China Agricultural University obtains.
No. 6 alfalfas of middle lucerne by the variety certification of the national committee for examination and approval of grass varieties, are registered as incubation new varieties on June 12nd, 2010, and kind certificate registration number is 422, and public Ke Cong China Agricultural University obtains.
Red careless No. 1 assorted russian fenugreek herb record date is on December 13rd, 2006, and cultivar registration number is 322, and public Ke Cong China Agricultural University obtains.
Visitor's alfalfa record date is on December 13rd, 2006, and cultivar registration number is 323, and public Ke Cong China Agricultural University obtains.
In document " the clover science production technology solution " book that the kinds such as CCID 5, CCID 7, CCID 10, three get profit, moral treasured, Saite, queen are published in Chinese agriculture publishing house, disclosed, the public can buy and obtain from market.
A kind of Alfalfa Breeding method that embodiment 1, cold plasma are processed
The Alfalfa Breeding method of utilizing cold plasma to process, can obtain output high, and quality better is nutritious, good palatability, has cold-resistant, drought-enduring, Salt And Alkali Tolerance, strong stress resistance, well developed root system, regeneration power are strong, and adaptability extensively waits the New alfalfa cultivars of characteristic trait, specifically comprises the steps:
One, the breeding material that collection meets breeding goal is as emphasis seed selection object
Concrete breeding material is No. 1 alfalfa of middle lucerne, No. 2 alfalfas of middle lucerne, No. 3 alfalfas of middle lucerne, No. 6 alfalfas of middle lucerne, red careless No. 1 assorted russian fenugreek herb, visitor's alfalfa, and the seed of the alfalfa variety such as CCID 5, CCID 7, CCID 10, three get profit, moral treasured, Saite, queen.
Two, the seed of the selected alfalfa variety of step 1 is placed in cold plasma processor, use high-octane material aggregation state, under the processing power of 1-500W, the breeding material of clover (seed of clover) is carried out to the unionized width of 15-20 second and penetrate processing, the necessary condition of processing is to take helium as working media, in vacuum sealing environment, carries out.
Three, the breeding material of cold plasma being processed is preserved under low temperature drying condition, begins sowing in good time waiting.
Four, with through the same batch of seeds of corresponding alfalfa variety any processing, that step 1 is selected, cultivate the clover (contrast) obtaining and compare, through the seed of cold plasma processing, cultivate the clover obtaining and realized the purification and rejuvenation of corresponding parent material, output raising, hybrid seed yield increase, quality improving and resistance enhancing.
(1) cold plasma is processed as shown in table 1 on the impact of the germination vigor of alfalfa seed and germination rate.
Germination vigor and the germination rate of alfalfa seed under the different cold plasma treatment conditions of table 1
The ratio that germination vigor in table 1 or germination rate improve represent breeding material that cold plasma processes with not through the comparing with a collection of breeding material of corresponding alfalfa variety any processing, that step 1 is selected, the ratio that germination vigor or germination rate improve.
Table 1 shows, when processing power is 20W, the processing time, while being 18s, the germination vigor of the seed of the clover of processing through cold plasma, germination rate improved maximum, than contrast, improve 10.01%-32.56% and 6.54%-14.55% respectively, realized the purification and rejuvenation of parent material.
(2) cold plasma is processed as shown in table 2 on the impact of clover plant height.
The plant height of clover under the different cold plasma treatment conditions of table 2
In table 2, the ratio that plant height improves represent breeding material that cold plasma processes with not through the comparing with a collection of breeding material of corresponding alfalfa variety any processing, that step 1 is selected, the ratio that plant height improves.
Table 2 shows, when processing power is 80W, the processing time, while being 18s, the plant height of the clover of processing through cold plasma improved maximum, than contrast, exceeds 21.25%-41.79%.
(3) cold plasma is processed as shown in table 3 on the impact of alfalfa hay rate of growth, fresh careless rate of growth.
The hay rate of growth of clover, fresh careless rate of growth under the different cold plasma treatment conditions of table 3
In table 3, hay or fresh careless rate of growth represent the breeding material of cold plasma processing and do not pass through comparing with a collection of breeding material of corresponding alfalfa variety any processing, that step 1 is selected, the rate of growth of hay or fresh grass.
Table 3 shows, when processing power is 20W, the processing time, while being 18s, the alfalfa hay rate of growth being obtained by the seed of processing through cold plasma, fresh careless rate of growth improved maximum, be respectively 22.16%-26.49% and the 18.67%-21.30% of contrast, the output that has realized parent material improves.
(4) cold plasma is processed the bear pods impact of number of clover as shown in table 4.
The number that bears pods of clover under the different cold plasma treatment conditions of table 4
In table 4, the number that bears pods improve breeding material that ratio represents that cold plasma processes with not through the comparing with a collection of breeding material of corresponding alfalfa variety any processing, that step 1 is selected, the ratio that the number that bears pods improves.
Table 4 shows, when processing power is 100W, 160W and 200W, processing time is while being 18s, the number that bears pods of the clover being obtained by the seed of processing through cold plasma improves 80.13%-113.5%, 78.54%-114.9% and 61.89%-82.80% than contrast respectively, and the hybrid seed yield of having realized parent material increases.
More than experiment shows, when processing power is within the scope of 1-500W, compared with the control, quality and the resistance of the clover being obtained by the seed of processing through cold plasma obviously strengthen.
Five, the breeding material of cold plasma being processed carries out adding generation and Field Screening, chooses target make a variation by generation.
Adding generation is exactly to exceed the breeding cycle of species itself and the reproductive process of breeding quantity, also only in the situation that of manual intervention, uses certain technological means (such as artificial pollination or insemination) just can reach.Adding generation is for shortening breeding cycle, and increasing actual production has important meaning.
Utilize DNA molecular marker technology to carry out assistant breeding, can differentiate variety of crops, assessment varietal purity, analyze agronomic trait gene, Isolation and Identification genetic stocks, determines homology of chromosome, detection to chromosome nonhomologous and chromosomal structural aberration, thus carry out assistant breeding.
Enzyme is the product of gene, is the result of gene expression.Enzyme difference is structurally mainly derived from gene, and Isozyme spectra of LDH is the phenotype of the molecular level after gene expression, and research Isozyme spectra of LDH just likely contacts the relation of gene, gene expression and metabolism, cell and whole phenotype.Because the similitude of isodynamic enzyme structure has been reacted interbiotic affiliation, so the classification that can be used for kind of Isozyme Analysis, isozyme data can be used as evaluation species, the important indicator of research classification, evolution, heredity and variation.Therefore, utilize Identification of Isozyme can carry out assistant breeding.
Individual plant 1st generation when plantation by through step 2, process and with not through same a collection of breeding material (seed) the subregion sowing simultaneously of corresponding alfalfa variety any processing, that step 1 is selected, carry out comparative trial, observation economical character and the coefficient of variation, adopt the means such as molecular labeling and Identification of Isozyme to carry out assistant breeding simultaneously, carry out individual plant selection, select good variation.2nd generation is carried out individual plant selection, selfed breeding.The 3rd generation and later each generation, can adopt individual plant, group, mixing system of selection according to the degree of fixation of strain, until be bred as the inbred line of good stable.
Six, the inbred line obtaining with step 5, the inbred line that the advantage of selecting is strong, meet breeding goal is as emphasis seed selection material, then selfing purification and rejuvenation.Purification and rejuvenation are in order to be good purity and the good strains of seeds that keeps alfalfa variety, can be applied to for a long time produce, the purification and rejuvenation of clover conventional variety, mainly adopt pod row method of purification (claiming again modified method of mixing selection), this method is simple, instant effect.
Select generally 2-3 days before clover pod maturation of pod link, in seed-breeding field, select pod.Selected pod must possess the typical proterties of this kind, robust plant simultaneously, and anosis worm harm, plant height, maturation all should be consistent.Each kind is selected the quantity of pod, should determine according to the demand of original seed.After gathering, carry out single pod evaluation, eliminate the bad single pod of a part; The elected pod threshing respectively of same processing, eliminates a part of seed according to seed properties after threshing again, and the seed of each processing packs a little paper bag into.The seed pouch that then same place should be selected is concentrated packing, and indicates and process title, properly preserves, and prevents infested or mouldy.
Plant comparing element, experimental plot adopts randomized arrangement, repeats for 3 times.Community area is 3m * 5m, and the amount of broadcasting is 3.0kg/hm
2, adopt the sowing of 40cm * 80cm wide-and narrow-row.Finish rear drawn field planting figure.Between breeding time, field management is timely, even, careful.In each breeding time, repeatedly screen, and assorted weak plant row is made a mark.When maturation, mix harvest-seed extraction, dry preservation.
Through the seed selection about 4 generations, reach inbred line stable like this, carry out variety comparative test and regional test, production demonstration and Quality Detection etc., carry out afterwards kind technical appraisement, breeding of new variety work completes.
By above step, obtain output high, quality better, nutritious, good palatability, has cold-resistant, drought-enduring, Salt And Alkali Tolerance, strong stress resistance, well developed root system, regeneration power are strong, and adaptability extensively waits the New alfalfa cultivars of characteristic trait, meets expection standard.
Claims (10)
1. the Alfalfa Breeding method that cold plasma is processed, comprises the steps:
(1) pending clover raw material are carried out to cold plasma processing, obtain the material after cold plasma is processed;
(2) in the material from cold plasma is processed, seed selection has the clover of objective trait.
2. method according to claim 1, is characterized in that: the condition that described cold plasma is processed is: take helium as working media, in vacuum sealing environment, the unionized width that under the processing power of 1-500W, clover raw material is carried out to 15-20s is penetrated processing.
3. method according to claim 1 and 2, it is characterized in that: described objective trait for as lower at least one: with without the comparing with a collection of clover raw material of any processing, alfalfa seed germinating capacity improves, clover plant height improves, alfalfa output improves and clover hybrid seed yield improves;
Described processing power is 10-400W, is specially 10-300W.
4. according to the arbitrary described method of claim 1-3, it is characterized in that: described processing power is 10-400W, processing time is while being 15-20s, with without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves and/or the plant height of clover improves and/or the output of clover improves and/or the hybrid seed yield of clover increases.
5. according to the arbitrary described method of claim 1-4, it is characterized in that: described processing power is 10-300W, processing time is while being 15-20s, with without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves and/or the plant height of clover improves and/or the output of clover improves and/or the hybrid seed yield of clover increases.
6. according to the arbitrary described method of claim 1-5, it is characterized in that: described processing power is 10-200W, the processing time is while being 15-18s, and without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
7. according to the arbitrary described method of claim 1-6, it is characterized in that: described processing power is 10-160W, processing time is 15-18s, or processing power is 100-300W, and the processing time is 18-20s, or processing power is 100-200W, processing time is 18s, or processing power is 100-160W, and the processing time is while being 18s, with without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
8. according to the arbitrary described method of claim 1-7, it is characterized in that: described processing power is 20W, the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the alfalfa seed germinating capacity after processing improves;
Described processing power is 80W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the plant height of the clover after processing improves;
Described processing power is 20W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the output of the clover after processing improves;
Described reason power is respectively 100W, 160W or 200W, and the processing time is while being 18s, and without the comparing with a collection of clover raw material of any processing, the hybrid seed yield of the clover after processing increases.
9. according to the arbitrary described method of claim 1-8, it is characterized in that: described alfalfa seed germinating capacity is alfalfa seed germination vigor and/or germination rate;
Described alfalfa output is presented as hay rate of growth and/or fresh careless rate of growth;
Described clover hybrid seed yield is presented as the number that bears pods;
The seed that described clover raw material are clover.
10. the application of the arbitrary described method of claim 1-9 in Alfalfa Breeding.
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