CN1579144A

CN1579144A - Method for culturing lily triploid

Info

Publication number: CN1579144A
Application number: CN 03153342
Authority: CN
Inventors: 郑思乡; 黄静; 张力
Original assignee: YUNNAN LEISEN SCIENCE AND TECHNOLOGY Co Ltd
Current assignee: YUNNAN LEISEN SCIENCE AND TECHNOLOGY Co Ltd
Priority date: 2003-08-11
Filing date: 2003-08-11
Publication date: 2005-02-16
Anticipated expiration: 2023-08-11
Also published as: CN1234265C

Abstract

The invention relates to a plant variety cultivating method, especially a method of cultivating lily polyploidy. 1. according to routine lily cultivating technique, selecting seed balls and jarovizing at low temperature to break dormancy to complete the jarovizing; 2. cultivating in a greenhouse or making field cultivation and management according to routine cultivating lily technique; 3. cultivating for 30-40days, then dripping 0.02-0.2% colchicines at 20 deg.C-25 deg.C; 4. according to the routine cultivating technique, processing the plants so that they will come into flower in 50-60 days, selecting the plants with extremely large stigmas and huge direct microscopic pollen particles to eliminate stamens so as to obtain 2n-oogamete containing stamens; 5. gathering normal pollens to hybridize with the obtained stamens, then according to routine cultivating technique, cultivating so that the fruits will swell fully in 45-60 days, thus able to gather them; 6. adopting routine triploid embryo in vitro culture method to be able to obtain the lily triploids.

Description

The triploid breeding method of lily

Technical field

The present invention relates to a kind of breeding method of plant variety, especially the breeding method of lily polyploid.

Technical background

Lily is Liliaceae (Liliaceae) lilium (Lilium) plant, and lily is big, rich color, and flower appearance grace, particularly oriental hybrid lily (Oriental hybrids) also have strong fragrance, and be very popular.Lily cut flowers is one of flowers at present with fastest developing speed in the world, and market prospects are wide.

Polyploid is one of approach of plant evolution, and polyploid has special quality because of its huge property, low pregnant property, resistance and the characteristics such as sterility, particularly triploid that overcome source far away hybridization and powerful nourish and grow advantage and favored by gardening breeding man.The existing report of lily polyploid research discovers that tetraploid plant is tall and big, has the material of more blooming.The lily breeding technique mainly is to adopt interspecific cross and intervarietal cross technology to cultivate new varieties.Adopt colchicine to handle scale and test-tube plantlet, can only obtain autotetraploid, can not obtain triploid.Obtaining triploid conventional method is with tetraploid and diploid hybrid, needs 5 years at least but the method obtains the lily required time of triploid, adopts the short period to obtain the triploid cultivation technology of lily and does not appear in the newspapers as yet.

Summary of the invention

Purpose of the present invention is intended to overcome the deficiencies in the prior art, provides a kind of and can obtain the triploid cultivation technology of lily in the short time.

The triploid breeding method of lily of the present invention is made up of following steps:

(1), by lily conventional cultivation technology, selecting Zhou Jing is the healthy and strong anosis kind ball of 14～18cm, through 2～5 ℃ of low temperature vernalization 90-120 days, breaking dormancy was finished vernalization, cultivates then in the matrix of sterilizing;

(2), in the greenhouse, cultivate, environmental condition is: 20～25 ℃ of temperature, and illumination 5～60,000 Lnx, air humidity is controlled at 50～60%; Perhaps carry out the field cultivation management with reference to the conventional cultivation technology;

(3), by the cultivation of above cultivation step after 30～40 days, plant height reaches 30～40cm, bud is under 20～25 ℃ of conditions with 0.02～0.2% colchicine in temperature when naked eyes are visible, adopts drop method to handle bud 2～3 days, with clear water cleaning treatment position;

(4) to the plant of above processing, culture technique is handled the back and was bloomed in about 50～60 days routinely, selects the column cap unusual loose and huge person of microscopy pollen grain that becomes, and stamen is removed the gynoecium that can obtain to contain the 2n oogamete;

(5), adopt normal pollen and above-mentioned gynoecium pollination hybridization, culture technique cultivation routinely then, hybridization back 45～60 days, fruit fully expands, and can gather;

(6), behind the above-mentioned fruit harvesting, adopt the cultured in vitro method of conventional triploid embryo, can obtain the lily triploid.

Breeding method of the present invention can obtain the lily triploid rapidly, produces the required time of triploid than the tetraploid of routine and diploid hybrid and shortens about 4 years; And the very strong advantage of nourishing and growing is arranged, have sterility and polyphyll; Adopting present technique to obtain triploid fast lays a good foundation for cultivating lily polyphyll WUHUAFEN (or few pollen) kind.

Embodiment

Selecting Zhou Jing is the healthy and strong anosis kind ball of 14～16cm, and by lily conventional cultivation technology, through 2～5 ℃ of low temperature vernalization 90 days, breaking dormancy was finished vernalization, cultivates then in the matrix of sterilizing; In environmental condition is 20～25 ℃ of temperature, illumination 5～60,000 Lnx, after cultivating 30～40 days in the greenhouse of air humidity 50～60%, plant height reaches 30～40cm, and bud is when naked eyes are visible, is under 20～25 ℃ of conditions with 0.02～0.2% colchicine in temperature, adopt drop method to handle bud 2～3 days, every morning 8～10 points, afternoon 2～4 each drop once, induce the 2n oogamete, use clear water cleaning treatment position again; To above plant, enhance field management, guarantees good growing environment, culture technique is handled afterwards and was bloomed in about 50～60 days routinely, and the plant of handling not is 100% morphs, and has only 9.52～25.8% aberration rate, and different cultivars differs greatly.The plant that only morphs just can produce the 2n gamete, the evaluation of 2n gamete is mainly differentiated by morphological observation and micrometering pollen grain size, and column cap becomes loose unusually and the huge person of microscopy pollen grain can be used as the Preliminary Identification index that the 2n gamete is induced success.Select the column cap unusual loose and huge person of microscopy pollen grain that becomes, stamen is removed the gynoecium that can obtain to contain the 2n oogamete;

With normal pollen and above-mentioned gynoecium pollination hybridization, bagging is listed, mark hybrid combination and date, and culture technique cultivation routinely then, hybridization back 45～60 days, fruit fully expands, and can gather; Behind the above-mentioned fruit harvesting, adopt the cultured in vitro method of following conventional triploid embryo, can obtain the lily triploid.

The cultured in vitro of triploid embryo: the low temperature that is placed on 5～7 ℃ behind the fruit harvesting was handled 6～8 days down, handled according to routine disinfection then; 75% alcohol is handled 30 seconds → 0.1%HgCl ₂(mercuric chloride) handles 10～12 minutes → aseptic washing 3 times → stripping seed or embryo is seeded to Ms+0.5～1.0mg/L 6-BA+1.0mg/LGA ₃On+0.1mg/LNAA the medium, 22～25 ℃ of cultivation temperature, illumination 1500～2000Lux, about embryo germination Cheng Miao more than 50% after 15～16 days, about embryo of about 50% is sprouted successively, and front and back need one-year age.

The evaluation of triploid lily: the lily seedling of above cultivation is not all to be triploid, obtaining triploid must identify by chromosome, chromosome is identified and is adopted F-BSG method or aceto-camine pressed disc method to carry out chromosome sectioning, the microexamination chromosome number, most plant chromosome numbers are 36 in this result of study, be triploid, i.e. 2n=3x=36, also having chromosome number is that 24 plant is that dliploid is 2n=2x=24.The tetraploid plant of also finding minority simultaneously is 2n=4x=48.The evaluation of trisome is the sign that this technology is succeedd.

Breeding of triploid lily: the conventional raising technology by the dliploid lily is carried out, the scale cultivation induced bundle on Ms+1.0～2.0mg/L6-BA+0.05～0.1mg/L NAA medium that strips the triploid lily is sprouted, successive transfer culture repeatedly on above-mentioned medium, at last with bud switching root induction on Ms+1.0mg/L NAA medium, additional 0.05% activated carbon, rooting efficiency is better.Test-tube plantlet was handled under 2～4 ℃ low temperature 40～56 days, generally handled about 47 days better, moved to leaf mould then: in the matrix of sand=1: 1, send behind the sprouting at height above sea level 2000～2400m, P in soil H value is that plant in 5～6 place.Strengthen the rich water quality management and the extermination of disease and insect pest, the small seed ball of gathering after leaf is withered.Small seed ball with breaking dormancy, carries out numerous ball second time 2～5 ℃ of following cold treatments 70～90 days then under above-mentioned height above sea level and soil condition.Promptly there is part seed ball can produce the commodity blossoming bulb, can breed triploid lily newtype.

Claims

The triploid breeding method of a kind of lily, form by following steps:

(1), by lily conventional cultivation technology, selecting Zhou Jing is the healthy and strong anosis kind ball of 14～18cm, through 2～5 ℃ of low temperature vernalization 90-120 days, breaking dormancy was finished vernalization, cultivates then in the matrix of sterilizing;
(2), in the greenhouse, cultivate, environmental condition is: 20～25 ℃ of temperature, and illumination 5～60,000 Lnx, air humidity is controlled at 500～60%; Perhaps carry out the field cultivation management with reference to the conventional cultivation technology;

It is characterized in that;
(3), by the cultivation of above cultivation step after 30～40 days, plant height reaches 30～40cm, bud is under 20～25 ℃ of conditions with 0.02～0.2% colchicine in temperature when naked eyes are visible, adopts drop method to handle bud 2～3 days, with clear water cleaning treatment position;
(4) to the plant of above processing, culture technique is handled the back and was bloomed in about 50～60 days routinely, selects the column cap unusual loose and huge person of microscopy pollen grain that becomes, and stamen is removed the gynoecium that can obtain to contain the 2n oogamete;
(5), adopt normal pollen and above-mentioned gynoecium pollination hybridization, culture technique cultivation routinely then, hybridization back 45～60 days, fruit fully expands, and can gather;
(6), behind the above-mentioned fruit harvesting, adopt the cultured in vitro method of conventional triploid embryo, can obtain the lily triploid.