CN1934934A - Efficient banana in vitro quick-breeding method - Google Patents
Efficient banana in vitro quick-breeding method Download PDFInfo
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Abstract
The present invention relates to a method for isolated fast propagation of high-quality seedling by using Brazil banana immature male flower as explant. Said method includes the following steps: collecting fresh banana male flower, using its immature portion, slicing and making tissue culture. The propagation coefficient of every slice explant is up to 25-30, and high-effective high-quality tissue culture seedling can be obtained.
Description
[technical field]
The present invention relates to utilize Brazilian banana prematurity male flower to breed the method for high quality seedling fast for explant exsomatizes.
[background technology]
The banana of broad sense comprises banana (Banana) and plantain (Plantain) two big classes.Banana and plantain belong to Musaceae (Musaceae) Musa (Musa) (Agricultural University Of South China chief editor. each opinion of pomology (south this). the agriculture .1993:104 of publishing house).Nearly all edible any of several broadleaf plants all obstructs any of several broadleaf plants (Musa balbisiana Colla) selfing by original wild sharp leaf any of several broadleaf plants (Musa acuminata Colla) and length or the filial generation evolution forms.Eat banana raw and mostly be triploid AAA greatly, promptly their chromosome is the offspring of selfing of sharp leaf any of several broadleaf plants or somatic mutation all from sharp leaf any of several broadleaf plants.Plantain also is a triploid, and chromosome set becomes AAB or ABB, and promptly their chromosome is from sharp leaf any of several broadleaf plants and long stalk any of several broadleaf plants.Except that triploid, also have dliploid AA, AB and tetraploid AAAA, BBBB, but the kind rareness (Samson JA.Tropical Fruits.London:longman, 1980:119).Cultivation banana (Musa spp.) is unisexuality triploid (AAA, AAB, ABB), has the height sterility.
Banana is the large-scale herbaceous monocotyledon of perennial evergreen, and subterranean stem is a thick bulb, and root, leaf, flower and stock suction bud grow thus.By the structural terminal bud germination of central cylindrical, constantly take out the leave sheet at first, stretch on the petiole, leaf sheath is right after and is false stem, forms bulb below the false stem, and every leaf sheath base resting bud can grow up to the suction bud, inhales bud and parent bulb and forms any of several broadleaf plants clump.The banana plant certain phase that grows, false stem growing point stop leaf development and carry out the reproduction differentiation, and locusta puts forth ears.
The cultivation banana mostly is triploid, parthenocarpy.A string fruit ear has fruit 4～18 combs, every comb has and really refers to 7～35, the comb fruit is diminished gradually by last, the any of several broadleaf plants fruit needs 65～170 days Zi blooming to results, high temperature season can be gathered in the crops in 60～90 days, low temperature season then need more than 120 days (the development south subtropics crop chief editor of office of the Ministry of Agriculture. Chinese tropical subtropical fruit tree. the .1998:49-50 of Chinese agriculture publishing house).
Banana all occupies important economic status in world's tropical fruit (tree) and Chinese tropical fruit (tree) production.According to statistics, world's banana production occupies the 3rd in fruit, be only second to grape and oranges and tangerines.According to the FAO statistics, more than 120 countries in the whole world (area) plantation banana in 2005, harvest area is 443.95 ten thousand hectares, gross yield reaches 7246.46 ten thousand tons, world's banana exported 1,594 ten thousand tons in 2004, and export value is the fruit of volume of world trade maximum near 5,000,000,000 dollars.China's national banana planting area in 2004 is about 28.5 ten thousand hectares, output reaches 6,290,000 tons, the output value is 9,400,000,000 yuan, national banana planting area was about 27.4 ten thousand hectares in 2005, output reaches 6,390,000 tons, and the output value surpasses 10,000,000,000 yuan, and the practitioner reaches more than 110 ten thousand, the banana industry has become China's agriculture pillar industry in south subtropics area, occupies important status in the social and economic development of hot-zone.
Traditional banana breeding has two kinds of methods, a kind of is directly to utilize the suction bud of maternal plant growth to breed, but this method is because the suction bud speed that produces is slow, reproduction coefficient is low, influence the growth of maternal plant, incompatible modern large-scale commodity production is eliminated gradually, has only the plantation of minority peasant family adopting at present; Another kind is to utilize to inhale the tissue culture technique that the gemmule stem manually carries out numerous bud, and this technology is the main means of sapling multiplication in the present banana production.This technology is China's researcher in the later stage eighties in last century because the exploitation of anti-season banana culture technique, and market increases high-quality bananas seedling demand, and market researches and develops the high quality seedling needs in order to satisfy.This The Application of Technology has played great progradation to the fast development of China's banana industry, has realized within a short period of time the improved seeds large tracts of land being promoted, and the large-scale commercial banana production is come true.But through nearly 20 years application, found that at present this method comes with some shortcomings aborning, the main problem that exists has:
1 subculture is counted height.Numerous bud tissue culture technology that banana is traditional can only reach about 2.5-3.0 at present because reproduction coefficient is low, needs repeatedly successive transfer culture so obtain a large amount of seedlings, generally all reaches successive transfer culture 10-12 generation, so could obtain high economic benefit.And tissue cultivating seedling is along with the increasing of subculture number, and seedling quality descends gradually and aberration rate raises, and the damage by disease and insect resistivity is reduced.But the producer often by subculture quantity increase increasing seedling output, therefore causes a large amount of seedlings inferior to come into the market in order to pursue high economic interests and to reduce cost in producing, and infringement banana peasant interests influence the banana industry development.
2 production cycles are long.The numerous tissue culture method of present banana bud probably needs 10-11 month time from gathering the suction bud to the commercial seedling in 12 generations, and the production cycle is long.
3 is easily malicious in spite of illness.The material that existing banana seedling production is adopted is the curved shot stem eye, and bulb grows in the underground, infects multiple germ easily, though screen sterilization through strict, but still have the danger of material band poison.For guaranteeing to produce seedling quality, the annual seedling of being produced will detect through virus, has increased the seedling production cost.
4 cause the maternal plant injury.The commercial in the market main sampling time of tissue culture seedlings of bananas concentrates on the annual 3-5 month, the banana maternal plant of this moment is in the high-speed rapid growth phase, the collection of sapling multiplication material can bring certain damage to the banana maternal plant, influences output and the fruit quality of maternal plant.
5 production cost height.Tissue culture seedlings of bananas production is the industry of a labour intensive, and reproduction coefficient is low, and the production cycle is long, and factory's labour's production cost accounts for more than 70% of whole production cost, and labour's production cost is too high.General produce 12 generation tissue cultivating seedling need time of the 10-11 month, consume great amount of manpower and and take a large amount of means of production, increased production cost.
The deficiency and the shortcoming that exist in the above banana group training mode of production, begun to restrict the quick healthy development of banana industry, also restricted the further upgrading of banana industry simultaneously, concrete manifestation has four aspects: first: the production cycle is long, market reaction is slow, can not tackle the market demand flexibly.Second: the high seedling quality that causes of subculture number descends, and brings the bad chain reaction in the production.The 3rd: the defective of method of drawing material has caused tissue culture seedlings of bananas detoxification fully, and draws materials then and may produce injury and to cause output to descend and the fruit quality reduction maternal plant.The the 4th: the production cost height.This tissue culture mode need consume a large amount of manpower and materials, the production cost height, and efficient is low.Therefore, develop a kind of new tissue culture technology and overcome above shortcoming, guarantee that the permanent sound development of banana industry has become one of banana production problem demanding prompt solution.And banana prematurity male flower tissue culture technique can remedy the defective of bulb tissue cultured seedling propagating method, provide safeguard for banana production provides the high-quality seedling.
Banana prematurity male flower tissue culture technique has the advantage of following six aspects:
1 reproduction coefficient height, with short production cycle.Male bud is an indefinite inflorescence, and its apical meristem has powerful former based system, the banana prematurity male flower tissue culture technology of our independent development, and reproduction coefficient can reach the level of 25-30.Utilize banana prematurity male flower section, the every strain male flower 20-24 sheet of can cutting into slices, each male flower section is through 25-30 bud of callus induction differentiation and bud formation, reproduction coefficient height.Single banana prematurity male flower induces a differentiation generation just can produce 500-720 strain banana seedlings by section.The tissue cultivating seedling production cycle of therefore producing equal number than the numerous bud method reduced in comparison of bud 3-4 month, the production cycle shortens greatly.
2 subculture numbers are low.Because the reproduction coefficient height so produce the tissue cultivating seedling of equal number, is compared with traditional tissue culture technology, banana prematurity male flower tissue culture technique only needs subculture 2-4 generation, greatly reduce the subculture number of group training, prevented effectively that therefore the bud that occurs because the subculture number is high is slim and frahile, growth potential is inhomogeneous, young shoot bleaches, serious minus green lacks vigor and vitality, and growth rate slows down, the rate of increase reduces, problems such as aberration rate height.Banana prematurity tissue cultivating seedling subculture number is low, and it is neat healthy and strong to emerge, and disease resistance is strong, fast growth, and aberration rate is low, and it is fast to take root.With respect to traditional tissue culture technology bigger advantage is arranged.
3 is not malicious in spite of illness.It is the growing point of banana prematurity male flower that this tissue culture technique adopts material, is the position of high-speed rapid growth, and by the natural tight bag quilt of bract, not malicious in spite of illness under field conditions (factors).
4 science, simple of drawing materials more do not injure the normal growth of the maternal plant of drawing materials.Compare with inhaling bud, the inflorescence tissue has characteristics such as drawing materials conveniently, sterilize easily, save provenance, is to reduce primary stage of inoculation to pollute, the good explant of raising success rate.
5 draw materials and improve output combines.In the actual process of drawing materials, utilize the bud that cutting off male flower cluster produced in the banana production, not only propagating materials can be obtained, but also output and the fruit quality of sampling maternal plant can be improved, realized drawing materials and the perfect adaptation of high yield technique.
6 inflorescences are easy to carry, help international germplasm and exchange, and to cultivating and introduce precocity, high yield, the famous and precious rare variety of high-quality, the improved seeds of particularly introducing foreign country especially have actual application value.
[summary of the invention]
A kind of summary of the invention of efficient banana breeding new method comprises that the prescription of explant selection, processing, evoked callus medium, the cultural method of evoked callus, the prescription of inductive differentiation medium, the cultural method of inducing differentiation and tissue cultivating seedling are urged contents such as root, domestication and transplanting.
1. explant selection
Choose the fresh banana male flower of banana cutting off male flower cluster phase
2. the processing of explant
Progressively divest banana male flower bract, to when male flower length is 9-10 centimetre, changeing people's superclean bench, in superclean bench, continue to divest banana male flower bract, be 2.0-2.5 centimetre extremely to male flower length, therefore the banana male flower at this position claims prematurity banana male flower owing to also do not reach maturity.The prematurity banana male flower at this position vertically on average is cut to two halves, and crosscut thickness is 1.2 millimeters thin slice again.
3. banana prematurity male flower section evoked callus is cultivated
The section of prematurity male flower is inoculated in MS+1.0mg/L vitamin h+100mg/L glutamine+0.2mg/L TDZ+0.2mg/L Zeatin+40g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.3,25-26 ℃, dark culturing.Look the brownization degree of section, changed a subculture in every 5-10 days, continuous culture 20-30 days, to producing callus.
4. differentiation of calli is cultivated
The banana prematurity male flower section that produces callus is inoculated in MS+4.0mg/L BA+0.5mg/L NAA+30g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.8.At 26-28 ℃, luminous intensity is to cultivate under the 1500lx condition, and every day, light was cultivated 12 hours, and dark culturing is 12 hours again.Changed a subculture in per 10 days, continuous culture 20-40 days, to differentiation generation banana seedling.
5. strong seedling culture
Forward the seedling of differentiation to MS+BA 3.0mg/L+NAA 0.3mg/L+30g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.8.At 26-28 ℃, luminous intensity is to carry out light under the condition of 1500-1700lx to cultivate, and changes a subculture in per 15 days.
6. culture of rootage
After strong seedling culture, banana seedlings is inoculated in MS+NAA0.5mg/L+ sucrose 30g/L+ fine jade 5.5g/L, on the medium of pH=5.8, under 28 ℃ of conditions, every day, illumination cultivation was 12 hours, and intensity is 1500lx, became the seedling of taking root to the root system that induces better quality in continuous culture 30-40 days.
7. the seedling of taking root domestication and transplanting
Take root seedling after 2-3 days natural daylight hardening, clean the root medium, transplant in nutritious bag,, be transplanted into the land for growing field crops field planting again at raise seedling in greenhouse 2-3 month.
Interpretation of result:
1 banana prematurity male flower is not with bacterium and virus under the state of self-sow, carefully divest the male flower bract at superclean bench, when being 2.0 centimetres, prematurity male flower length cuts into slices, owing to grow not step of body outward through sterilizing, reduce injury, greatly improved outer the going out more and differentiation rate of body of growing.
2 choose the fresh banana prematurity male flower section of banana cutting off male flower cluster phase, and brownization of section that obtains is few, goes out sooner, goes out the more, and differentiation smoothly.Must choose fresh banana male flower, otherwise serious brownization of cutting into slices, go out less and the differentiation difficulty, greatly influence the acquisition of tissue culture seedlings of bananas.
3 slice thicknesses also have very significant effects to going out more and breaking up, too thin brownization of section back poor growth, and dead easily, healing rate is low; It is too thick to cut into slices, and when thickness surpassed 2 millimeters, it is fast that speed is expanded in section, and increasing fast is one, also being difficult for more differentiation, and slice thickness goes out in the time of 1.2 millimeters more and differentiation reaches best in the test.
4 utilize banana prematurity male flower section, the every strain male flower 20-24 sheet of can cutting into slices, and each male flower section is by callus induction differentiation and bud formation 25-30, the reproduction coefficient height.Single banana prematurity male flower induces a differentiation generation just can produce 500-720 strain banana seedlings by section, and this is much higher than the reproduction coefficient of the numerous bud tissue culture technology of banana bud.
5 because the reproduction coefficient height so produce the tissue cultivating seedling of equal number, is compared with the numerous bud tissue culture technology of present bud, and banana prematurity male flower tissue culture technique only needs subculture 2-4 for just passable, greatly reduces the subculture number of group training.Because the subculture number is very low, prevented effectively that therefore the bud that occurs because the subculture number is high is slim and frahile and uneven, young shoot bleaches, and serious minus green lacks the life vigor, and growth rate slows down, and the rate of increase reduces, problems such as aberration rate height.Banana prematurity tissue cultivating seedling subculture number is low, and it is neat healthy and strong to emerge, and disease resistance is strong, fast growth, and aberration rate is low, and it is fast to take root.Tissue culture technology with respect to the numerous bud of bud has great advantage.
[embodiment]
1 material
The fresh cutting off male flower cluster male flower of Brazilian banana that gather the China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute proving ground.
2 methods
2.1 the processing of material
The banana male flower of fresh collection is progressively divested bract, to when male flower length is 9-10 centimetre, changeing people's superclean bench.Continue to divest banana male flower bract in superclean bench, to being 2.0 centimetres to male flower length, vertically on average be cut to two halves, crosscut thickness is 1.2 millimeters thin slices again.
2.2 callus culture
Section is inoculated in MS+1.0mg/L vitamin h+100mg/L glutamine+0.2mg/L TDZ+0.2mg/L Zeatin+40g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.3,25-26 ℃, dark culturing, look the brownization degree of section, change a subculture in every 5-10 days, 20-30 days continuous culture time, produce callus.
2.3 differentiation culture
The banana prematurity male flower section of growth callus is inoculated in MS+4.0mg/L BA+0.5mg/L NAA+30g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.8,26-28 ℃, light is cultivated, intensity is 1500lx, changed a subculture in per 10 days, 20-40 days continuous culture time, differentiation banana seedling.
2.4 strong seedling culture
Forward the seedling of differentiation to MS+BA 3.0mg/L+NAA 0.3mg/L+30g/L sucrose+5.5g/L agar, on the solid culture medium of pH=5.8,26-28 ℃, light is cultivated, and luminous intensity is 1500-1700lx, per 15 days replacing one subcultures.
2.5 urging root cultivates
After strong seedling culture, banana seedlings is inoculated in MS+NAA0.5mg/L+ sucrose 30g/L+ fine jade 5.5g/L, pH=5.8,28 ℃, every day, illumination cultivation was 12 hours, and luminous intensity is 1500lx.Then induced the root system of better quality, and became the seedling of taking root in continuous culture 30-40 days.
Seedling domestication and transplanting 2.6 take root
Take root seedling after 2-3 days natural daylight temperature hardening, clean the root medium, transplant in nutritious bag,, be transplanted into the land for growing field crops field planting again at raise seedling in greenhouse 2-3 month.
Claims (4)
1 one kinds of efficient banana breeding new methods, its inventive features is explant selection and processing, evoked callus culture medium prescription and method, inductive differentiation medium prescription and method etc.
2 choose and handle according to described its explant of claim 1 and are characterised in that: the fresh banana male flower of choosing the banana cutting off male flower cluster phase, progressively divesting male flower bract to male flower length is 2.0-2.5 centimetre, the prematurity male flower at this position vertically on average is cut to two halves, and crosscut thickness is 1.2 millimeters thin slice again.
3 are characterised in that according to described its evoked callus culture medium prescription of claim 1 and method: MS+1.0mg/L vitamin h+100mg/L glutamine+0.2mg/L TDZ+0.2mg/L Zeatin+40g/L sucrose+5.5g/L agar, the solid culture medium of pH=5.3.The section that claim 2 is chatted is inoculated on this solid culture medium, and 25-26 ℃, dark culturing was changed a subculture in every 5-10 days, 20-30 days continuous culture time, produces callus.
4 its inductive differentiation medium prescriptions according to claim 1 and method are characterised in that: MS+4.0mg/L BA+0.5mg/LNAA+30g/L sucrose+5.5g/L agar, the solid culture medium of pH=5.8.The callus that claim 3 is produced is inoculated on this solid culture medium, and 26-28 ℃, every day, illumination cultivation was 12 hours, and intensity is 1500lx, changes a subculture in per 10 days, 20-40 days continuous culture time, differentiation banana seedling.
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| CN103283597A (en) * | 2013-05-13 | 2013-09-11 | 中国热带农业科学院海口实验站 | Method for improving banana immature male flower embryonic callus induction success rate |
| CN103444535A (en) * | 2013-09-05 | 2013-12-18 | 广东省农业科学院果树研究所 | Novel method for increasing tissue culture and rapid propagation coefficients of banana |
| CN103651149A (en) * | 2014-01-08 | 2014-03-26 | 中国热带农业科学院热带生物技术研究所 | Screening method for cold resistant banana germplasm |
| CN103988743A (en) * | 2014-04-21 | 2014-08-20 | 黄少伟 | Banana planting method |
| CN107047301A (en) * | 2017-03-22 | 2017-08-18 | 黄庆辉 | A kind of banana strong seedling culture base for adding cumarin |
| CN107232055A (en) * | 2017-06-14 | 2017-10-10 | 中国热带农业科学院海口实验站 | A Xi any of several broadleaf plants breeding methods based on improved culture medium |
| CN108476981A (en) * | 2018-03-27 | 2018-09-04 | 中国热带农业科学院热带生物技术研究所 | A kind of screening technique of anti-blight banana germplasm |
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1062599C (en) * | 1996-12-23 | 2001-02-28 | 中国科学院遗传研究所 | Tissue culture for banana |
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2006
- 2006-10-27 CN CNB2006101426613A patent/CN100463600C/en not_active Expired - Fee Related
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| CN103444535A (en) * | 2013-09-05 | 2013-12-18 | 广东省农业科学院果树研究所 | Novel method for increasing tissue culture and rapid propagation coefficients of banana |
| CN103651149A (en) * | 2014-01-08 | 2014-03-26 | 中国热带农业科学院热带生物技术研究所 | Screening method for cold resistant banana germplasm |
| CN103651149B (en) * | 2014-01-08 | 2016-06-22 | 中国热带农业科学院热带生物技术研究所 | A kind of screening technique of cold resistant banana germplasm |
| CN103988743A (en) * | 2014-04-21 | 2014-08-20 | 黄少伟 | Banana planting method |
| CN107047301A (en) * | 2017-03-22 | 2017-08-18 | 黄庆辉 | A kind of banana strong seedling culture base for adding cumarin |
| CN107232055A (en) * | 2017-06-14 | 2017-10-10 | 中国热带农业科学院海口实验站 | A Xi any of several broadleaf plants breeding methods based on improved culture medium |
| CN108476981A (en) * | 2018-03-27 | 2018-09-04 | 中国热带农业科学院热带生物技术研究所 | A kind of screening technique of anti-blight banana germplasm |
| CN108476981B (en) * | 2018-03-27 | 2024-05-03 | 中国热带农业科学院热带生物技术研究所 | Screening method of banana germplasm with resistance to fusarium wilt |
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