CN101897295B - Method for breeding multiploid Chinese tallow tree new product by combination of hybridization and embryo culture - Google Patents

Abstract

The invention discloses a method for breeding a multiploid Chinese tallow tree new product by combination of hybridization and embryo culture. The invention adopts a technical scheme that a hybrid is prepared by hybridizing different types of excellent Chinese tallow tree varieties; the hybrid is subjected to immature embryo callus Initiation, colchicine doubling, renewal cultivation, seedling differentiation, differentiated seedling rooting culture and plant transplantation to form a tetraploid Chinese tallow tree hybrid; and a new Chinese tallow tree tetraploid line can be determined by a method for identifying the morphological characteristics of diploid Chinese tallow tree, observation of the stomatalcell chloroplast number and comparison of chromosome number. In the method, the new multiploid Chinese tallow tree line with strong stress resistance and high oil yield is cultured by sufficiently playing the double advantages of hybridization and multiploid, and contributes to improving the oil yield of raw materials for preparing biodiesel, relieving energy crisis, improving environmental protecting capability and reducing carbon consumption index.

Description

The combine method of seed selection polyploid Chinese tallow tree new of a kind of hybridization and embryo culture

Technical field

The present invention relates to the method for economic tree breeding, particularly a kind of hybridization and embryo culture combine the method for seed selection polyploid Chinese tallow tree new, through the method for improved seeds hybridization and the good tetraploid Chinese tallow tree new of multiploid induction seed selection.

Background technology

Along with fossil energy faces exhaustion day by day, people begin worldwide to seek alternative energy source.The develop energy plant replaces fossil energy to hold out broad prospects.Kind of energy-source plant surplus listing 60 after China Forestry Science Research Institute's research, wherein the woody oleiferous plants plant is its important species.It both can be used as solid fuel, can refine bio-crude oil again.U.S. Nobel laureate Ka Er professor just developed first artificial petroleum plant, and obtained the harvest of per hectare 120-140 bucket bio-crude oil in 1984.According to the medium-term and long-term ENERGY PLANNING of China; The renewable energy utilization total amount will be equivalent to 600,000,000 tons of standard coals to the year two thousand twenty whole nation, wherein greatly develop energy-source plant, and especially oil plants will be to the structural adjustment of China energy; Reduce greenhouse gas emission, aspect performance significant role such as preserve the ecological environment.

Chinese tallow tree (Sapium sebiferum Roxb) is a kind of desirable woody oleiferous plants energy-source plant, be China distinctive with oil tea, tung oil tree and walnut and be called one of seeds of China four big woody oleiferous plants plants.It is the Euphorbiaceae deciduous tree, fruit oblate spheroid, pitchy, cracking when ripe.Seed black is outward by Chinese wax; Seed oil content about 40% is called Chinese catalpa oil; Wax is called skin oil by oil content about 35%, and is fine with its refinement bio-crude oil effect.But the per unit area yield of China Chinese tallow tree is very low, and per mu yield only tens is arrived the level of hundreds of kilogram well below more than 1000 kilogram of U.S.'s per mu yield.The Chinese tallow tree new seed selection of China also only is confined to the collection seed selection stage to farm variety.Therefore, carry out different cultivars hybridization and the polyploid breeding of Chinese tallow tree, performance hybrid and double dominancy of polyploid, for breeding high-yield Chinese tallow tree breeding, it is very important to improve output.

Summary of the invention

The objective of the invention is to propose a kind of method of utilizing hybridization and polyploid to combine the seed selection Chinese tallow tree new.Solve existing Chinese tallow tree and produce the main landrace that rely on, the present situation that oil production is low is through enforcement of the present invention; Will be for obtaining strong stress resistance; The polyploid Chinese tallow tree new that oil production is high provides technology and breeding material, for improving the raw-material oil production of preparation biodiesel, alleviating energy crisis; Improve the environmental protection ability, reduce the carbon consumption index and contribute.

The present invention realizes like this.Its technical scheme is:

1), be that maternal shank Chinese tallow tree is cooked male parent with the grape Chinese tallow tree, obtain crossbreed in the Chinese tallow tree Sheng phase pollination hybridization of blooming;

2), the callus induction of immature Chinese tallow tree hybrid embryo is cultivated; Take to hybridize the hybrid fruit that grew 25-40 days the back; Under aseptic condition, stripping out the hybrid embryo inserts in the callus inducing medium and to cultivate 30 days-35 days; The callus inducing medium composition is MS+NAA 0.1mg/l-1.0mg/l+6-BA0.5mg/l-3.0mg/l+sucrose 3.0%-5.0%+agar0.75%-1.2%, pH6.0;

3), double to handle, the Chinese tallow tree hybrid embryo callus of vigorous growth is placed on doubles that induced chromosome doubled to handle 36-56 hour in the inducing culture; Double the inducing culture based component for not adding callus inducing medium+colchicine 350mg/L-1000mg/l of agar;

4), recover to cultivate, the Chinese tallow tree hybrid embryo callus after doubling to handle is changed over to recovered in the callus inducing medium to cultivate 7-10 days;

5), differentiation and seedling emergence; Chinese tallow tree hybrid embryo callus after recover cultivating changed in the inductive differentiation medium cultivated 10 days-30 days; Break up the seedling that sprouts; The inductive differentiation medium composition is MS+6BA1mg/l-3.5mg/l+NAA 0.1mg/l-0.7mg/l+sucrose 2.0%-3.5%+agar0.75%-1.2%, pH6.0;

6), the differentiation seedling rooting is cultivated; The bud seedling plant division of Chinese tallow tree hybrid embryo callus differentiation is inserted root media; In test tube, breaking up seedling rooting cultivated 15-20 days; The culture of rootage based component is MS+IBA0.7mg/l-1.5mg/l+NAA0.1mg/l-0.5mg/l+sucrose 2.0%-3.0%+agar0.75%-1.2%, pH6.0;

7), plant transplants, and will take root, the above Chinese tallow tree test-tube plantlet of plant height 3.0cm is planted in the sterilization compost of 1/3 fine sand+1/3 fine earth+1/3 vermiculite, shades and preserves moisture, treat seedling length behind the 10.0cm height again balled transplanting in the field;

8), morphology identifies, according to the plant height of tetraploid Chinese tallow tree than the short 10%-25% of dliploid Chinese tallow tree, the thick 25%-35% of blade, the characteristics of the close 15%-25% of stipes are tentatively distinguished tetraploid and dliploid Chinese tallow tree plant;

9), cytology and chromosome number purpose are identified; According to the stomatal cell of the vacuum side of blade of tetraploid Chinese tallow tree than the diplontic 35%-45% that lacks; Big 30%-40%; Tetraploid stomatal cell number of chloroplast is than the diplontic 65%-85% that manys, and the tip of a root and stem apex cell chromosome number tetraploid Chinese tallow tree are diplontic diploidy numbers, and combining form is learned characteristics and confirmed the tetraploid Chinese tallow tree.

The beneficial effect of the invention:

Through enforcement of the present invention; The double dominant of giving full play to hybridization and polyploid is the acquisition strong stress resistance; The polyploid Chinese tallow tree new that oil production is high provides technology and breeding material, for improving the raw-material oil production of preparation biodiesel, alleviating energy crisis; Improve the environmental protection ability, reduce the carbon consumption index and contribute.

Description of drawings:

Fig. 1, be two kinds of good Chinese tallow tree grape variety Chinese tallow trees (left side) and shank Chinese tallow tree (right side) and crossbreed plant thereof (in) picture, hybrid shows tangible hybrid vigour;

Fig. 2,3,4,5 handles callus differentiation and seedling emergence picture for the embryo culture of Chinese tallow tree hybrid becomes the callus colchicine.

Embodiment

Embodiment 1

1. select good Chinese tallow tree landrace summer shop grape Chinese tallow tree to do female parent, the shank Chinese tallow tree of realizing is a male parent;

2. in Chinese tallow tree 10 of mid-June Sheng phase: the 30-14 of blooming: 30 pollination hybridization;

3. take to hybridize the hybrid fruit that grew 35 days the back; Under aseptic condition, strip out the hybrid embryo and insert MS+NAA 0.5mg/l+6-BA 0.5mg/l+sucrose 3.0%+agar0.75%; Cultivated 30 days in the callus inducing medium of compositions such as pH6.0, evoked callus forms;

4. treat the vigorous growth of Chinese tallow tree hybrid embryo callus, change the callus inducing medium+colchicine 500mg/l that does not add agar when presenting faint yellow half graininess over to, handle the 48hr induced chromosome and double;

5. the callus after will doubling to handle changes callus filter over to and leads and recover in the medium to cultivate 8 days;

6. will double callus and change inductive differentiation medium over to, MS+6BA 1mg/l+NAA 0.5mg/l+sucrose 2.0%+agar 0.75% cultivated 30 days among the pH6.0, broke up the seedling that sprouts;

With the bud seedling plant division of differentiation insert MS+ [BA 0.7mg/l+NAA0.1mg/l+sucrose2.0%+agar 0.75%, culture of rootage is 15 days in the pH6.0 root media;

8. will take root, the above Chinese tallow tree test-tube plantlet of plant height 3.0cm is planted in the sterilization compost of 1/3 fine sand+1/3 fine earth+1/3 vermiculite, shades and preserves moisture, treat seedling length behind the 10.0cm height again balled transplanting in the field, grow;

According to the plant height of tetraploid Chinese tallow tree than the short 10%-25% of dliploid Chinese tallow tree, the thick 25%-35% of sheet, the characteristics of the close 15%-25% of stipes, the morphology that doubles the Chinese tallow tree seedling is identified, tentatively distinguishes tetraploid and dliploid Chinese tallow tree plant;

According to the stomatal cell of the vacuum side of blade of tetraploid Chinese tallow tree than the diplontic 35%-45% that lacks; Big 30%-40%; Tetraploid stomatal cell number of chloroplast is than the diplontic 65%-85% that manys; The tip of a root and stem apex cell chromosome number tetraploid Chinese tallow tree are diplontic diploidy numbers, and the cytology and the chromosome number purpose that double the Chinese tallow tree seedling identify that combining form is learned characteristics and confirmed the tetraploid Chinese tallow tree.

Claims (1)

1. hybridization and the embryo culture method of seed selection polyploid Chinese tallow tree new that combines is characterized in that incubation step is:

1), be that female parent, shank Chinese tallow tree are done male parent with the grape Chinese tallow tree, obtain crossbreed in the Chinese tallow tree Sheng phase pollination hybridization of blooming;

2), immature Chinese tallow tree hybrid embryo being carried out callus induction cultivates; Take to hybridize the hybrid fruit that grew 25-40 days the back; Under aseptic condition, stripping out the hybrid embryo inserts and cultivates 30-35 days in the callus inducing medium; The callus inducing medium composition is MS+NAA 0.5mg/l+6-BA 0.5mg/l+sucrose 3.0%+agar0.75%, pH6.0;

3), double to cultivate, the Chinese tallow tree hybrid embryo callus of vigorous growth is placed on doubles that induced chromosome doubled to handle 36-56 hour in the inducing culture; Double the inducing culture based component for not adding callus inducing medium+colchicine 500mg/l of agar;

4), recover to cultivate, the Chinese tallow tree hybrid embryo callus after doubling to handle is changed over to recovered in the callus inducing medium to cultivate 7-10 days;

5), differentiation and seedling emergence; Chinese tallow tree hybrid embryo callus after recover cultivating changed in the inductive differentiation medium cultivated 10 days-30 days; Break up the seedling that sprouts, the inductive differentiation medium composition is MS+6BA 1mg/l+NAA0.5mg/l+sucrose 2.0%+agar 0.75%, pH6.0;

6), the differentiation seedling rooting is cultivated; The bud seedling plant division of Chinese tallow tree hybrid embryo callus differentiation is inserted root media; In test tube, break up seedling rooting and cultivated 15-20 days, the culture of rootage based component is MS+IBA0.7mg/l+NAA0.1mg/l+sucrose 2.0%+agar 0.75%, pH6.0:

7), plant transplants, and will take root, the above Chinese tallow tree test-tube plantlet of plant height 3.0cm is planted in the sterilization compost of 1/3 fine sand+1/3 fine earth+1/3 vermiculite, shades and preserves moisture, treat seedling length behind the 10.0cm height again balled transplanting in the field;

8), morphology identifies, according to the plant height of tetraploid Chinese tallow tree than the short 10%-25% of dliploid Chinese tallow tree, the thick 25%-35% of blade, the characteristics of the close 15%-25% of stipes are tentatively distinguished tetraploid and dliploid Chinese tallow tree plant;

9), cytology and chromosome number purpose are identified; According to the stomatal cell of the vacuum side of blade of tetraploid Chinese tallow tree than the diplontic 35%-45% that lacks; Big 30%-40%; Tetraploid stomatal cell number of chloroplast is than the diplontic 65%-85% that manys, and the tip of a root and stem apex cell chromosome number tetraploid Chinese tallow tree are diplontic diploidy numbers, and combining form is learned characteristics and confirmed the tetraploid Chinese tallow tree.

Images