CN103651149A - Screening method for cold resistant banana germplasm - Google Patents
Screening method for cold resistant banana germplasm Download PDFInfo
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Abstract
The invention belongs to the field of banana callus irradiation mutation breeding, and particularly relates to a screening method for cold resistant banana germplasm. According to the method, the cold resistant banana germplasm can be quickly and efficiently screened. The screening method for the cold resistant banana germplasm comprises the following steps of: the irradiation treatment of the immature male flower callus of bananas, the low-temperature screening of the callus, the low-temperature screening of a differentiated bud, the low-temperature screening of a banana seedling and banana cold resistant rooting seedling acclimation and transplantation. According to the method, because the low-temperature screening technology is adopted for three times, especially, the low-temperature screening technology is adopted during the period of the male flower induction callus, so that the screening efficiency of the cold resistant mutation germplasm is greatly improved, and the period for culturing a cold resistant banana variety is shortened.
Description
Technical field
The invention belongs to banana callus radiaction mutation field, be specifically related to a kind of screening technique of cold-resistant banana germplasm.
Technical background
Banana is typical tropical and subtropical zone fruit tree, is distributed widely in the torrid zone and South Subtropical Area of China.China is one of world banana main product state, and planting area is distributed in the provinces and regions such as Guangdong, Hainan, Guangxi, Fujian, Yunnan and Taiwan.Due to the variation of weather conditions, the banana planting of the provinces such as China Guangdong, Guangxi, Yunnan is threatened by cold damage often in recent years, causes drop in production over a large area and have no harvest, and causes serious economic loss to vast banana orchard worker.Although in production, for cold damage, set up some preventive measure, for more serious cold damage, its preventive effect or undesirable, so the cold-resistant kind of seed selection banana is the fundamental solution that solves research on chilling injury in banana problem.The banana major part of cultivation is triploid at present, and parthenocarpy has sterility highly, cannot pass through the cold-resistant new varieties of conventional hybridization seed selection, therefore utilizes
60co-gamma Rays mutagenesis banana, cultivates banana mutant, and from mutant, screening has the new germ plasm of cold-resistant sudden change, then cultivation acquisition new varieties are one of important channels of the cold-resistant breeding of banana.
Banana radioinduction is carried out to cold-resistant breeding research both at home and abroad at present and report that document is less, in the selection of radioinduction material, the main banana that adopts is inhaled bud, banana group training bud or Banana Seedlings, its method major defect is to need mutant material to be processed many, and the screening cycle is long, efficiency is low.It is radiomutation material that the screening technique of a kind of cold-resistant banana germplasm of the present invention is chosen banana prematurity male flower callus, adopt three low temperature triage techniqueses of the screening of callus low temperature, the screening of Bud Differentiation low temperature, the screening of Banana Seedlings low temperature, improve the screening effeciency of cold-resistant sudden change germplasm, shortened the cycle of cultivating the cold-resistant kind of banana.
Summary of the invention
A kind of summary of the invention of screening technique of cold-resistant banana germplasm comprises that the cold-resistant contents such as seedling domestication and transplanting of taking root of irradiation treatment, the screening of callus low temperature, the screening of Bud Differentiation low temperature, the screening of Banana Seedlings low temperature, the banana of banana prematurity male flower callus form.
(1) irradiation treatment of banana prematurity male flower callus
The callus warp of banana prematurity male flower induction
60co-gamma Rays is processed, and callus radiation absorbed dose is 60-70Gy.
(2) callus low temperature screening
Through the callus of irradiation treatment, proceed to climatic cabinate and carry out dark culturing, it is 14 ℃, 10 ℃ two thermogrades that climatic cabinate temperature is set, and adopts the principle of low temperature after first high temperature, and each Temperature Treatment time cycle is 10 days, carries out low temperature screening.Recovery temperature to 26 ℃ progressively after 10 ℃ of low temperature screening stages finish, selects the callus surviving after low temperature screening to break up cultivation.Callus subculture medium is the solid culture medium of MS+1.0mg/L vitamin h+100mg/L glutamine+0.2mg/L TDZ+0.2mg/L Zeatin+40g/L sucrose+5.5g/L agar pH=5.3, within every 10 days, changes a subculture.
(3) differentiation of callus is cultivated
The callus surviving after low temperature screening is inoculated on the differentiation solid culture medium of MS+4.0mg/L BA+0.5mg/L NAA+30g/L sucrose+5.5g/L agar pH=5.8.At 26 ℃, luminous intensity is to cultivate under 1500lx condition, and every daylight is cultivated 12 hours, dark culturing 12 hours.Within every 10 days, change and once break up solid culture medium, within continuous culture 20-30 days, produce Bud Differentiation.
(4) Bud Differentiation low temperature screening
Bud Differentiation proceeds to climatic cabinate, it is 1500lx that climatic cabinate luminous intensity is set, every daylight is cultivated 12 hours, dark culturing 12 hours, it is 14 ℃, 10 ℃, 7 ℃ three thermogrades that climatic cabinate temperature is set, the principle that adopts low temperature after first high temperature, each Temperature Treatment time cycle is 14 days, carries out low temperature screening.Recovery temperature to 26 ℃ progressively after 7 ℃ of low temperature screening stages finish, selects the Bud Differentiation that survives after low temperature screening, at 26 ℃, cultivates 21 days, and Bud Differentiation grows into Banana Seedlings.
(5) Banana Seedlings low temperature screening
Banana Seedlings is proceeded to climatic cabinate.It is 1700lx that climatic cabinate luminous intensity is set, every daylight is cultivated 18 hours, dark culturing 6 hours, it is 14 ℃, 10 ℃, 7 ℃, 4 ℃ four thermogrades that climatic cabinate temperature is set, adopt the principle of low temperature after first high temperature, each Temperature Treatment time cycle is 10 days, carries out the screening of Banana Seedlings low temperature.Recovery temperature to 26 ℃ progressively after 4 ℃ of low temperature screening stages finish, the Banana Seedlings that selection survives after low temperature screening, Banana Seedlings is inoculated on the root media of MS+NAA0.5mg/L+ sucrose 30g/L+ fine jade 5.5g/L pH=5.8, continuous culture 30 days, induce the root system of better quality, become the cold-resistant seedling of taking root of banana.
(6) cold-resistant take root seedling domestication and transplanting of banana
The cold-resistant seedling of taking root of banana, after the natural daylight hardening of 2-3 days, is cleaned root medium, transplants in nutritious bag, at raise seedling in greenhouse 2-3 month, then is transplanted into Field planting, as cold-resistant banana germplasm, preserves.
Embodiment
The present invention is further described in connection with example below.
1 material
China tropic Agriculture Academy Sciences tropic Biotechnology Research Institute's Biotechnological Research on Bananas group provides banana prematurity male flower callus.
2 banana prematurity male flower callus radioinductions are processed
Banana prematurity male flower callus warp
60co-gamma Rays is processed, and callus radiation absorbed dose is 60-70Gy, and irradiation treatment is completed by Institute of Nuclear and Biotechnology, Sichuan Academy of Agriculture Science.
3 callus low temperature screenings
The callus of process irradiation treatment proceeds to climatic cabinate and carries out dark culturing.It is 14 ℃, 10 ℃ two thermogrades that climatic cabinate temperature is set, and adopts the principle of low temperature after first high temperature, and each Temperature Treatment time cycle is 10 days, carries out low temperature screening.Recovery temperature to 26 ℃ progressively after 10 ℃ of low temperature screening stages finish, selects the callus surviving through low temperature screening to break up cultivation.Callus subculture medium is the solid culture medium of MS+1.0mg/L vitamin h+100mg/L glutamine+0.2mg/L TDZ+0.2mg/L Zeatin+40g/L sucrose+5.5g/L agar pH=5.3, within every 10 days, changes a subculture.
The differentiation of 4 callus is cultivated
The callus that screening survives through low temperature is inoculated on the solid culture medium of MS+4.0mg/L BA+0.5mg/L NAA+30g/L sucrose+5.5g/L agar pH=5.8.26 ℃ of temperature, under luminous intensity 1500lx condition, to cultivate, every daylight is cultivated 12 hours, dark culturing 12 hours.Within every 10 days, change a subculture, continuous culture 20-30 days, to producing Bud Differentiation.
5 Bud Differentiation low temperature screenings
Bud Differentiation proceeds to climatic cabinate.It is 1500lx that climatic cabinate luminous intensity is set, and every daylight is cultivated 12 hours, dark culturing 12 hours, it is 14 ℃, 10 ℃, 7 ℃ three thermogrades that climatic cabinate temperature is set, the principle that adopts low temperature after first high temperature, each Temperature Treatment time cycle is 14 days, carries out low temperature screening.Recovery temperature to 26 ℃ progressively after 7 ℃ of low temperature screening stages finish, selects the Bud Differentiation that survives after low temperature screening, at 26 ℃, cultivates 21 days, and Bud Differentiation grows into Banana Seedlings.
6 Banana Seedlings low temperature screenings
Banana Seedlings is proceeded to climatic cabinate.It is 1700lx that climatic cabinate luminous intensity is set, every daylight is cultivated 18 hours, dark culturing 6 hours, it is 14 ℃, 10 ℃, 7 ℃, 4 ℃ four thermogrades that climatic cabinate temperature is set, adopt the principle of low temperature after first high temperature, each Temperature Treatment time cycle is 10 days, carries out the Screening Treatment of Banana Seedlings low temperature.Recovery temperature to 26 ℃ progressively after 4 ℃ of low temperature screening stages finish, the Banana Seedlings that selection survives after low temperature screening, Banana Seedlings seedling is inoculated on the medium of MS+NAA0.5mg/L+ sucrose 30g/L+ agar 5.5g/L pH=5.8, continuous culture becomes the cold-resistant seedling of taking root of banana to the root system that induces better quality in 30 days.
7 bananas cold-resistant take root seedling domestication and transplanting
The cold-resistant seedling of taking root of banana, after the natural daylight hardening of 2-3 days, is cleaned root medium, transplants in nutritious bag, at raise seedling in greenhouse 2-3 month.
Character observation and Determination of Physiological And Biochemical Indices after the cold-resistant transplantation of seedlings of taking root of 8 bananas
1. transplanting survival rate is added up
Transplanting survival rate is added up in the cold-resistant transplantation of seedlings of taking root of banana in nutritious bag after 2 months be 51.47%, and the contrast banana seedlings transplanting survival rate that does not experience low temperature screening is 98.67%, and the more difficult adaptation normal temperature of plant medium recovery growth through low temperature screening are described.
2. plant height and the number of blade are measured
Plant height and the number of blade are to describe the important indicator of Banana Seedlings growth rate speed.Average plant height is measured in the cold-resistant transplantation of seedlings of banana after 2 months be 9.60 centimetres, and the average number of blade is 7.83.The contrast banana seedlings that does not experience low temperature screening is transplanted and measured average plant height after 2 months is 20.10 centimetres, and the average number of blade is 9.33, illustrates that the cold-resistant seedling growth rate filtering out is slower.
3. the mensuration of cold-resistant seedling low temperature stress physiological and biochemical index
The cold-resistant transplantation of seedlings of taking root of banana after 3 months, is positioned over climatic cabinate in nutritious bag, and it is 1700lx that climatic cabinate luminous intensity is set, and every daylight is cultivated 18 hours, dark culturing 6 hours, and it is 7 ℃ that climatic cabinate temperature is set, low temperature stress 2 days.Measure peroxidase (POD), catalase (CAT) and malonaldehyde (MDA) content.Result shows that the cold-resistant seedling plant of banana peroxidase (POD) enzyme activity is 1.4 times of control groups; The cold-resistant seedling plant of banana catalase (CAT) enzyme activity is 1.78 times of control groups; The cold-resistant seedling plant of banana malonaldehyde (MDA) content is only control group 32.3%, shows that the cold-resistant seedling plant of banana has stronger cold tolerance.
Claims (1)
1. a screening technique for cold-resistant banana germplasm, its feature comprises the following steps:
1), the irradiation treatment of banana prematurity male flower callus
The callus warp of banana prematurity male flower induction
60co-gamma Rays is processed, and callus radiation absorbed dose is 60-70Gy;
2), callus low temperature screening
The callus of process irradiation treatment proceeds to climatic cabinate and carries out dark culturing, it is 14 ℃, 10 ℃ two thermogrades that climatic cabinate temperature is set, adopt the principle of low temperature after first high temperature, each Temperature Treatment time cycle is 10 days, carry out low temperature screening, recovery temperature to 26 ℃ progressively after 10 ℃ of low temperature screening stages finish, selects the callus surviving after low temperature screening to break up cultivation;
3), the differentiation of callus is cultivated
The callus surviving after low temperature screening is inoculated on differentiation solid culture medium, within continuous culture 20-30 days, produces Bud Differentiation;
4), Bud Differentiation low temperature screening
Bud Differentiation proceeds to climatic cabinate, it is 1500lx that climatic cabinate luminous intensity is set, every daylight is cultivated 12 hours, dark culturing 12 hours, it is 14 ℃, 10 ℃, 7 ℃ three thermogrades that climatic cabinate temperature is set, adopt the principle of low temperature after first high temperature, each Temperature Treatment time cycle is 14 days, carry out low temperature screening, recovery temperature to 26 ℃ progressively after 7 ℃ of low temperature screening stages finish, the Bud Differentiation that selection survives after low temperature screening, cultivates 21 days at 26 ℃, and Bud Differentiation grows into Banana Seedlings;
5), Banana Seedlings low temperature screening
Banana Seedlings is proceeded to climatic cabinate, it is 1700lx that climatic cabinate luminous intensity is set, every daylight is cultivated 18 hours, dark culturing 6 hours, it is 14 ℃ that climatic cabinate temperature is set, 10 ℃, 7 ℃, 4 ℃ of four thermogrades, adopt the principle of low temperature after first high temperature, each Temperature Treatment time cycle is 10 days, carry out the screening of Banana Seedlings low temperature, recovery temperature to 26 ℃ progressively after 4 ℃ of low temperature screening stages finish, the Banana Seedlings that selection survives after low temperature screening, Banana Seedlings is inoculated on root media, continuous culture 30 days, induce the root system of better quality, become the cold-resistant seedling of taking root of banana,
6), cold-resistant take root seedling domestication and transplanting of banana
The cold-resistant seedling of taking root of banana, after the natural daylight hardening of 2-3 days, is cleaned root medium, transplants in nutritious bag, at raise seedling in greenhouse 2-3 month, then is transplanted into Field planting, as cold-resistant banana germplasm, preserves.
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105052735A (en) * | 2015-09-07 | 2015-11-18 | 镇江瑞繁农艺有限公司 | Method for obtaining cold-resistant collard plant |
| CN105145237A (en) * | 2015-07-31 | 2015-12-16 | 王振华 | Plantation method for bananas |
| CN112293248A (en) * | 2020-11-30 | 2021-02-02 | 中国热带农业科学院海口实验站 | Chemical mutagenesis method for inducing and increasing mutation range of banana |
| CN113142055A (en) * | 2021-04-29 | 2021-07-23 | 广西壮族自治区农业科学院 | In-vitro proliferation preservation method for banana germplasm resources |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105145237A (en) * | 2015-07-31 | 2015-12-16 | 王振华 | Plantation method for bananas |
| CN105145237B (en) * | 2015-07-31 | 2018-06-19 | 宁波远志立方能源科技有限公司 | A kind of implantation methods of banana |
| CN105052735A (en) * | 2015-09-07 | 2015-11-18 | 镇江瑞繁农艺有限公司 | Method for obtaining cold-resistant collard plant |
| CN112293248A (en) * | 2020-11-30 | 2021-02-02 | 中国热带农业科学院海口实验站 | Chemical mutagenesis method for inducing and increasing mutation range of banana |
| CN112293248B (en) * | 2020-11-30 | 2021-12-21 | 中国热带农业科学院海口实验站 | Chemical mutagenesis method for inducing and increasing mutation range of banana |
| CN113142055A (en) * | 2021-04-29 | 2021-07-23 | 广西壮族自治区农业科学院 | In-vitro proliferation preservation method for banana germplasm resources |
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