CN107172976B - Improved sugarcane hybrid seed production method - Google Patents

Improved sugarcane hybrid seed production method Download PDF

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CN107172976B
CN107172976B CN201710506308.7A CN201710506308A CN107172976B CN 107172976 B CN107172976 B CN 107172976B CN 201710506308 A CN201710506308 A CN 201710506308A CN 107172976 B CN107172976 B CN 107172976B
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parent
liquid
culture medium
stem
hybridization
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CN107172976A (en
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雷敬超
高丽花
张保青
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Sugarcane Research Institute of Guangxi Academy of Agricultural Sciences
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Sugarcane Research Institute of Guangxi Academy of Agricultural Sciences
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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/02—Methods or apparatus for hybridisation; Artificial pollination ; Fertility
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01F—PROCESSING OF HARVESTED PRODUCE; HAY OR STRAW PRESSES; DEVICES FOR STORING AGRICULTURAL OR HORTICULTURAL PRODUCE
    • A01F25/00—Storing agricultural or horticultural produce; Hanging-up harvested fruit
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01H—NEW PLANTS OR NON-TRANSGENIC PROCESSES FOR OBTAINING THEM; PLANT REPRODUCTION BY TISSUE CULTURE TECHNIQUES
    • A01H1/00—Processes for modifying genotypes ; Plants characterised by associated natural traits
    • A01H1/04—Processes of selection involving genotypic or phenotypic markers; Methods of using phenotypic markers for selection
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05B—PHOSPHATIC FERTILISERS
    • C05B17/00—Other phosphatic fertilisers, e.g. soft rock phosphates, bone meal
    • C—CHEMISTRY; METALLURGY
    • C05—FERTILISERS; MANUFACTURE THEREOF
    • C05G—MIXTURES OF FERTILISERS COVERED INDIVIDUALLY BY DIFFERENT SUBCLASSES OF CLASS C05; MIXTURES OF ONE OR MORE FERTILISERS WITH MATERIALS NOT HAVING A SPECIFIC FERTILISING ACTIVITY, e.g. PESTICIDES, SOIL-CONDITIONERS, WETTING AGENTS; FERTILISERS CHARACTERISED BY THEIR FORM
    • C05G3/00—Mixtures of one or more fertilisers with additives not having a specially fertilising activity
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/14—Fungi; Culture media therefor
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
    • C12N1/20—Bacteria; Culture media therefor

Abstract

The invention relates to the technical field of agriculture, in particular to an improved sugarcane hybrid seed production method, which comprises the following specific steps: (1) parent breeding; (2) marking parents; (3) collecting flower spikes; (4) detecting pollen fertility; (5) preparing and combining; (6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, selecting 2-4 nodes on a stem section which is 200-250 cm below the base of the spica, removing leaf sheaths, wrapping with a nutrient medium, and carrying out rapid root promotion treatment for 7-15 days; (7) artificial pollination; (8) after-ripening of seeds: after hybridization, discarding the ear and stem of the male parent, transferring the female parent into an after-ripening room, and checking the effect of root promotion treatment; (9) and (5) harvesting the seeds. The method has the advantages of low cost, simple operation, avoiding errors caused by handwriting labels when the spica is collected, overcoming the problems of difficult stem wrapping, labor cost, spica cost and the like of the traditional sugarcane hybrid seed production, improving the utilization rate of the spica and the seed setting rate, and the like.

Description

Improved sugarcane hybrid seed production method
[ technical field ] A method for producing a semiconductor device
The invention relates to the technical field of agriculture, in particular to an improved sugarcane hybrid seed production method.
[ background of the invention ]
Sugarcane is the most important sugar crop in China, and the sucrose accounts for about 80% of the total sugar yield. Sexual hybridization is the basis of sugarcane breeding, and is a breeding mode with the widest application and obvious effect for variety improvement and updating and ensuring the sustainable development of the sucrose industry. After the sugarcane blooms, the success of hybridization is ensured, the quality of hybrid seeds is improved, and the method is the basis of sugarcane hybrid seed production. Currently, the hybridization methods used are: first, the adjacent row planting method of parents requires that the florescence of the parents is consistent and is difficult to realize. In the second method, the stem is cultured by sulfurous acid for the male parent, and the conservation of the female parent is strict in the later maturing stage so as to prevent the flower ears from withering and death, so that the obtained seeds have poor quality. Thirdly, the female parent takes roots automatically, and the male parent adopts a sulfurous acid stem-raising method, so that the female parent is inconvenient to move, and only can be hybridized outdoors, and the environmental influence is large. And fourthly, carrying out high-pressure rooting by using the stem wrapping of the female parent, and carrying out sulfurous acid stem culture on the male parent, wherein the method is the same as the third method, because the sugarcane stem grows to have roots, the nutrient supply is good, the maturing rate is high, the seed quality is good, but the booting condition of the female parent is required to be observed in a field, once the booting condition is found, the female parent is wrapped by using a nutrient medium, and after more fibrous roots grow at the stem wrapping position, the stem is cut off and then is transported back to a hybridization greenhouse for hybridization. The fourth method is currently used in most breeding farms, but there are many problems with this method. Firstly, the time selection of the phimosis is early, the flower spike is not opened, the nutrient medium of the phimosis is dry, and the fibrous root which just grows out is withered. Secondly, the female parent is determined by experience, but the climate is different every year, the environmental impact is different, and the same parent pollen fertility is different between years. Thirdly, the selection of the number of phimosis sometimes dozens of phimosis processed by one parent, but several spica are actually used, which causes great waste. Fourthly, in the booting stage, parents are 3-5 m high, and the phimosis can be completed by stairs, so that the workload is very large. Fifthly, after the phimosis roots, the height of the female parent is fixed, and after the phimosis roots enter a hybridization cage, the hanging height of the male parent is considered by completely wringing the brain juice. Therefore, there is a need to provide a new method for crossing sugarcane, which can reduce the workload and improve the seed setting rate.
[ summary of the invention ]
The invention aims to: aiming at the problems, the invention provides a hybridization method with low cost and simple operation, which avoids errors caused by handwriting labels when the spikes are collected, overcomes the problems of difficult stem wrapping, labor cost, spike cost and the like of the traditional sugarcane hybridization seed production, and improves the utilization rate of the spikes and the seed setting rate.
In order to achieve the purpose, the technical scheme adopted by the invention is as follows:
an improved sugarcane cross breeding method comprises the following specific steps:
(1) parent breeding: the method comprises the following steps of (1) sowing the easy-to-flower parent in spring of the year, sowing the difficult-to-flower parent in advance compared with the easy-to-flower parent, and stopping fertilizing in the last 6 months of each year;
(2) marking parents: making corresponding parent bar code labels for each parent to carry out distinguishing marking, and suspending the labels at parent planting places in 10 months each year;
(3) collecting flower spikes: the parents have the scions 1/3-1/2, the parents can be collected when a small amount of spikelets are bloomed at the tops, the parents are cut from the base parts of the sugarcane stalks, the longest state of the sugarcane stalks is kept, most of leaves are cut off, a handheld scanning printer is used for scanning a suspended parent bar code label plate, 2 new bar codes with the same name as that in the step (2) are printed, the 1 st part of the bar codes is pasted on the cut parents, 3-8 branches of flowering stalks of the parents are collected and collected in a storage box, the corresponding 2 nd part of the bar codes are attached, the sugarcane stalks cut at the base parts are transported back to a hybridization greenhouse, the spikes are sprayed with clear water and are inserted into a clear water stem-culturing pool for conservation in sequence;
the stem cultivating pool is divided into two rows, each row is divided into a plurality of small cells, and a position number bar code clamping groove area is arranged on the outer side of each small cell;
scanning the parent bar code and the position bar code of the position of the parent by using a scanner, processing the parent bar code and the position bar code by using a processor, and displaying the counted and collected spica number and position in a display;
(4) and (3) detecting pollen fertility: randomly selecting 2 florescent spikelets close to the florescent spikelets from the flowering spikelets retrieved in the step (3), wherein each spikelet contains 3 anthers, selecting 3 effective florets in total, obtaining 18 anthers in total, placing the anthers on a glass slide, adding an I-KI solution for dyeing, mashing the anthers by using a glass rod, detecting the pollen fertility under a microscope, and determining the paternal and maternal characteristics of the anthers;
(5) preparing and combining: according to the collected spica and the hybridization plan, making a hybridization combination which can be configured on the same day, printing a combination label plate, and placing a female parent and a male parent in the combination into the same hybridization cage; firstly fixing the position of the male parent, controlling the height of the male parent to be 30-50cm lower than the top end of the hybridization cage, fixing a stem-cultivating solution bottle with the volume of 5-10L at the corresponding position of the hybridization cage, cutting off part of internode tissues at an old incision when the lower end of the male parent is collected, and quickly inserting the cut part into the stem-cultivating solution bottle containing 5L and 150ppm of sulfurous acid solution; placing the female parent in the middle of a hybridization cage, placing the female parent below and placing the male parent above, overlapping 1/3 male and female parents, fixing a stem-nourishing solution bottle with the volume of 5-10L at the corresponding position of the hybridization cage, quickly cutting off part of internode tissues at an old incision when the lower end of the female parent is collected by a sharp knife, quickly inserting the internode tissues into the stem-nourishing solution bottle containing 5L and 150ppm of sulfurous acid solution, and hanging the combined label plate at the middle part of the sugarcane stem of the female parent;
(6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, selecting 2-4 nodes on a stem section which is 200-250 cm below the base of the spica, removing leaf sheaths, wrapping with a nutrient medium, and carrying out rapid root promotion treatment for 7-15 days;
(7) artificial pollination: artificial pollination is carried out every 1 hour when 8-10 am, and only the male parent flower spikes are knocked to ensure that pollen falls onto the female parent flower spikes; in the pollination period, if the flowering period of the male parent is ended in advance, the male parent is supplemented in time until the flowering of the female parent is basically ended, and the temperature in a hybridization greenhouse is controlled to be 25-28 ℃ and the relative humidity is controlled to be 75-85% in the artificial pollination period;
(8) after-ripening of seeds: discarding the ear stems of the male parents after hybridization, transferring the female parents into an after-ripening chamber, checking the effect of root promotion treatment, sheathing the whole flower ears downwards by a yarn bag, putting the combined label plate into the yarn bag, and fastening the lower end to prevent the label plate from falling off;
the method for checking the root promoting treatment effect comprises the following steps:
if more fibrous roots are generated, cutting off the fibrous roots 2-3cm below the phimosis, removing the wrapped film, and putting the phimosis into flowing clear water for culturing until the seeds are mature;
if no fibrous root can be produced, cutting off the internode tissue of 5-10cm lower end of the female parent with a sharp knife, quickly inserting into a barrel containing 20L and 150ppm of sulfurous acid solution, cutting off 2-3cm below phimosis after more fibrous root is produced, removing the wrapped film, and putting into flowing clear water for culturing until the seed is mature;
(9) harvesting seeds: when the seeds at the top of the female parent are white and flocculent and the spikelets fall off by more than two thirds, cutting off the spikes, putting the spikes into a drying room for drying treatment, then removing yarn bags, putting the seeds into waterproof paper bags, and storing the seeds for a long time at the temperature of between 15 ℃ below zero and 21 ℃ below zero;
the conditions of the drying treatment are as follows: drying at 11 deg.C and 13% humidity for 4 days;
further, the nutrient medium is prepared from the following raw materials in parts by weight: 0.1-0.5 part of indolebutyric acid, 0.1-0.5 part of naphthylacetic acid, 0.5-1.6 parts of rooting powder, 1-5 parts of alcohol, 0.3-0.7 part of compound amino acid injection, 10-25 parts of modified ceramsite powder, 10-20 parts of coconut coir, and 5-10 parts of compound microbial fertilizer;
the compound microbial fertilizer is prepared by mixing the following raw materials in percentage by weight: 1-3% of bacillus licheniformis liquid, 2-3% of bacillus megaterium liquid, 1-2% of yellow mould liquid, 1-3% of bacillus mucilaginosus liquid, 2-3% of bacillus thuringiensis liquid, 1-3% of trichoderma harzianum liquid, and the balance of base materials are supplemented to 100%;
wherein the effective bacterium content of the bacillus licheniformis liquid is 3 × 108-5×108The effective bacterium content of the bacillus megaterium liquid is 2 × 109-3×109The effective bacterium content of the yellow mold bacterium liquid is 4 × 109-6×109The effective bacterium content of the bacillus mucilaginosus liquid is 2 × 108-6×108The effective bacteria content of Bacillus thuringiensis liquid is 3 × 108-5×108The effective bacterium content of the trichoderma harzianum liquid is 1 × 10 per mL9-3×109Per mL;
the base material comprises peat, ground phosphate rock and waste sugar residues according to the weight ratio of 2-4:2: 2-3.
Further, the nutrient content of the nutrient medium is, by mass, 55% -65% of organic matters, 3% -4% of ammonium nitrogen, 0.1% -0.5% of nitrate nitrogen, 10% -20% of total nutrients, 5% -8% of available phosphorus and 8% -15% of available potassium.
Further, the bacillus licheniformis liquid is prepared by placing the strain of bacillus licheniformis in Na2SeO30.08-0.1g、CH3COONa 3-5g、NaHCO35-8g、(NH4)2SO46-8g、NH4Cl 2-3g、K2HPO43-4g、KH2PO40.2-0.5g、NaCl 0.5-0.6g、MgSO40.1-0.3g、CaCl20.2g of yeast extract, 15g of peptone 5-10g and 1L of distilled water, and culturing in a culture medium with pH adjusted to 7.3 with acetic acid to obtain Bacillus licheniformis with effective bacteria content of 3 × 108-5×108One by mL, namely;
the bacillus megaterium liquid is prepared by placing the strain of bacillus megaterium in Na2SeO30.1-0.2g、CH3CH2COONa1.5-2.5g、(NH4)2SO41.0-1.5g、K2HPO40.6-0.8g、KH2PO40.1-0.3g、MgSO40.2-0.25g、NaCl0.1-0.15g、CaCl20.2-0.3g of yeast extract, 1.5-2.0g of yeast extract and 1000mL of distilled water are prepared into a culture medium, and the culture medium is cultured for 3-5d to obtain the culture medium with the effective bacteria content of 2 × 109-3×109The dosage is one/mL.
Further, the yellow mold liquid is specifically prepared by inoculating yellow mold strains to a PDA culture medium slant for activated culture, then transferring to a PDA liquid culture medium for liquid culture, and then transferring to a broth culture medium for amplification culture until the effective bacteria content is 4 × 109-6×109One by mL, namely;
the trichoderma harzianum liquid is characterized in that trichoderma harzianum strains are inoculated on a PDA culture medium inclined plane for activated culture, then transferred into a PDA liquid culture medium for liquid culture, and then transferred into a broth culture medium for amplification culture until the effective bacteria content is 1 × 109-3×109The dosage is one/mL.
Further, the Bacillus mucilaginosus is prepared by placing strains of the Bacillus mucilaginosus in Na2SeO30.1-0.15g, peptone 5-10g, ferment3-5g of mother cream and KH2PO41.0-2.0g、MgSO4Culturing in 1.5-2.5g culture medium containing wort 150mL and distilled water 1000mL for 3-5d to obtain effective bacteria content of 2 × 108-6×108One by mL, namely; the bacillus thuringiensis liquid is prepared by inoculating bacillus thuringiensis strain to broth agar culture medium slant for slant activation training, culturing for 10-20 days, and transferring to culture medium composed of diammonium citrate 2.5-2.8g, peptone 30g, yeast extract 0.5-1.0g, and CH3CH2COONa 2.5-3.5g、KH2PO4Culturing in culture medium composed of 1.0-2.0g, MgSO40.8-1.2 g, wort 100mL, and distilled water 1000mL for 3-5d to obtain extract with effective bacteria content of 3 × 108-5 × 108/mL.
Further, the modified ceramsite powder is prepared by micronizing ceramsite, adding the micronized ceramsite into a mortar, adding olive oil into the mortar, grinding the mixture for 20-30 seconds, and irradiating the mixture for 5-8 seconds by gamma rays.
Further illustratively, in step (2), the parent barcode label contains the following information: parent name, bar code corresponding to the parent name, parent source, flowering sequence, past paternity and pollen fertility.
To further illustrate, in step (5), the combination tag includes the following data: female parent name, female parent position, male parent name, male parent position, hybrid cage number, preparation date and combination number.
In summary, due to the adoption of the technical scheme, the invention has the beneficial effects that:
1. by applying the invention, the parent name can be accurately recorded, and handwriting errors can be prevented; the sex of the parents is accurately judged, and invalid hybrid combinations are reduced; the utilization rate of the parents is improved, the workload of phimosis is reduced, the labor and the financial resources are saved, and the indoor centralized management is facilitated; the fibrous roots growing after stem wrapping are beneficial to the absorption of nutrients of the sugarcane stems, the final setting rate is high, and the seed quality is good.
2. The nutrient medium provided by the invention contains complete nutrient substances, can promote the sugarcane stalks to grow fibrous roots after stem wrapping more quickly, has more fibrous roots and thicker roots, has a large amount of activated beneficial microorganisms on the fibrous roots, and can ensure that the survival rate of the sugarcane stalks after stem wrapping by the nutrient medium is high.
[ description of the drawings ]
FIG. 1 is a state diagram after 10 days of the root-promoting treatment of the present invention;
FIG. 2 is a schematic representation of the present invention after being wrapped with a nutritional substrate;
FIG. 3 is a schematic diagram of a parent of the present invention after pasting a parent barcode;
FIG. 4 is a schematic view of the yarn-on-cover bag of the present invention collecting seeds;
fig. 5 is a hand held scanning printer scanning a suspended parent bar code label plate printing a bar code of the same name.
[ detailed description ] embodiments
In order to make the technical means, the creation features, the achievement purposes and the effects of the invention easy to understand, the invention is further described with the embodiment. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Example 1:
an improved sugarcane cross breeding method comprises the following specific steps:
(1) parent breeding: the method comprises the following steps of (1) sowing the easy-to-flower parent in spring of the year, sowing the difficult-to-flower parent in advance compared with the easy-to-flower parent, and stopping fertilizing in the last 6 months of each year;
(2) marking parents: making a corresponding parent bar code label for each parent to distinguish and mark, and suspending the label at a parent planting place in the last 10 months each year, wherein the parent bar code label comprises the following information: parent name, bar code corresponding to the parent name, parent source, flowering sequence, past paternity and pollen fertility;
(3) collecting flower spikes: cutting off the base part of the sugarcane stem, keeping the longest state of the sugarcane stem, cutting off most leaves, scanning a suspended parent bar code label by using a handheld scanning printer, printing 2 new bar codes with the same name as that in the step (2), pasting the 1 st part of bar codes on the cut-off parent, collecting 3 flowering stalks of the parent, storing the flowering stalks in a storage box and attaching the corresponding 2 nd part of bar codes, transporting the sugarcane stems cut off at the base part back to a hybridization greenhouse, spraying flower ears with clear water, and sequentially inserting the flower ears into a clear water stem culture pond for conservation;
the stem cultivating pool is divided into two rows, each row is divided into a plurality of small cells, and a position number bar code clamping groove area is arranged on the outer side of each small cell;
scanning the parent bar code and the position bar code of the position of the parent by using a scanner, processing the parent bar code and the position bar code by using a processor, and displaying the counted and collected spica number and position in a display;
(4) and (3) detecting pollen fertility: randomly selecting 2 florescent spikelets close to the florescent spikelets from the flowering spikelets retrieved in the step (3), wherein each spikelet contains 3 anthers, selecting 3 effective florets in total, obtaining 18 anthers in total, placing the anthers on a glass slide, adding an I-KI solution for dyeing, mashing the anthers by using a glass rod, detecting the pollen fertility under a microscope, and determining the paternal and maternal characteristics of the anthers;
(5) preparing and combining: according to the collected spica and the hybridization plan, a configurable hybridization combination in the same day is made, and a combination label is printed, wherein the combination label comprises the following data: female parent name, female parent position, male parent name, male parent position, hybridization cage number, preparation date and combination number, and then placing the female parent and the male parent in the combination in the same hybridization cage; firstly fixing the position of the male parent, controlling the height of the male parent to be 30cm lower than the top end of the hybridization cage, fixing a stem-cultivating solution bottle with the volume of 5L at the corresponding position of the hybridization cage, cutting off part of internode tissues at an old incision when the lower end of the male parent is collected, and quickly inserting the cut part into the stem-cultivating solution bottle containing 5L and 150ppm of sulfurous acid solution; placing the female parent in the middle of a hybridization cage, placing the female parent below and placing the male parent above, overlapping 1/3 male and female parents, fixing a stem-nourishing solution bottle with the volume of 5L at the corresponding position of the hybridization cage, quickly cutting off part of internode tissues at an old incision when the lower end of the female parent is collected by a sharp knife, quickly inserting the internode tissues into the stem-nourishing solution bottle containing 5L and 150ppm sulfurous acid solution, and hanging the combined label plate at the middle part of the sugarcane stem of the female parent;
(6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, 2 nodes are selected from a stem section which is 200cm below the base of the spica, the leaf sheath is removed, the stem section is wrapped by a nutrient medium, and the rapid root promoting treatment is carried out for 7 days;
(7) artificial pollination: artificial pollination is carried out every 1 hour when 8-10 am, and pollen is floated on the female parent spica only by shaking the male parent spica; in the pollination period, if the flowering period of the male parent is ended in advance, the male parent is supplemented in time until the flowering of the female parent is basically ended, and the temperature in a hybridization greenhouse is controlled to be 25 ℃ and the relative humidity is controlled to be 75% in the artificial pollination period;
(8) after-ripening of seeds: discarding the ear stems of the male parents after hybridization, transferring the female parents into an after-ripening chamber, checking the effect of root promotion treatment, sheathing the whole flower ears downwards by a yarn bag, putting the combined label plate into the yarn bag, and fastening the lower end to prevent the label plate from falling off;
the method for checking the root promoting treatment effect comprises the following steps:
if more fibrous roots are generated, cutting off the fibrous roots 2cm below the phimosis, removing the wrapped film, and putting the film into flowing clear water for culturing until the seeds are mature;
if no fibrous root is produced, rapidly cutting the internode tissue of 5cm at the lower end of the female parent by using a sharp knife, rapidly inserting the internode tissue into a barrel containing 20L of a sulfurous acid solution of 150ppm to produce more fibrous roots, cutting off the internode tissue 2cm below the phimosis, removing a wrapped film, and putting the internode tissue into flowing clear water for culturing until the seeds are mature;
the gauze bag is made of transparent gauze materials, one end of the gauze bag is sealed, the other end of the gauze bag is open, the diameter of the gauze bag is 40cm, the length of the gauze bag is 80-100cm, and the open end of the gauze bag is provided with a lock catch.
(9) Harvesting seeds: when the seeds at the top of the female parent are white and flocculent and the spikelets fall off by more than two thirds, cutting off the spikes, putting the spikes into a drying room for drying treatment, then removing yarn bags, putting the seeds into waterproof paper bags, and storing the seeds for a long time at the temperature of-21 ℃;
the conditions of the drying treatment are as follows: drying at 11 deg.C and 13% humidity for 4 days;
the nutrient medium is prepared from the following raw materials in parts by weight, wherein the nutrient medium contains 65% of organic matters, 4% of ammonium nitrogen, 0.5% of nitrate nitrogen, 11% of total nutrients, 5% of available phosphorus and 14.5% of available potassium: 0.1 part of indolebutyric acid, 0.1 part of naphthylacetic acid, 0.5 part of rooting powder, 1 part of alcohol and 0.3 part of compound amino acid injection, wherein ceramsite is subjected to ultramicro grinding, then the ceramsite is added into a mortar, olive oil is added into the mortar, the mixture is ground for 20 seconds, and then 10 parts of modified ceramsite powder, 10 parts of coconut husk and 5 parts of compound microbial fertilizer are obtained by irradiating 5 seconds with gamma rays.
The compound microbial fertilizer is prepared by mixing the following raw materials in percentage by weight: 1% of bacillus licheniformis liquid, 2% of bacillus megaterium liquid, 1% of yellow mold liquid, 1% of jelly-like bacillus liquid, 2% of bacillus thuringiensis liquid and 1% of trichoderma harzianum liquid, and the balance being base materials which are supplemented to 100%, wherein the base materials comprise peat, ground phosphate rock and waste sugar residues according to the weight ratio of 2:2: 2;
wherein the Bacillus licheniformis solution is prepared by placing strain of Bacillus licheniformis in Na2SeO30.08g、CH3COONa3g、NaHCO35g、(NH4)2SO46g、NH4Cl 2g、K2HPO43g、KH2PO40.2g、NaCl 0.5g、MgSO40.1g、CaCl20.2g of yeast extract, 15g of peptone and 1L of distilled water, and culturing in a culture medium prepared by adjusting the pH value to 7.3 with acetic acid to obtain the bacillus licheniformis with the effective bacteria content of 3 × 108One by mL, namely;
wherein the Bacillus megaterium solution is prepared by placing Bacillus megaterium strain in Na2SeO30.1g、CH3CH2COONa1.5g、(NH4)2SO41.0g、K2HPO40.6g、KH2PO40.1g、MgSO40.2g、NaCl 0.1g、CaCl20.2g of yeast extract, 1.5g of yeast extract and 1000mL of distilled water are cultured for 3d to obtain the effective bacteria content of 2 × 109One by mL, namely;
wherein the yellow mould liquid is prepared by inoculating yellow mould strain onto PDA culture medium slant for activation culture, transferring into PDA liquid culture medium for liquid culture, transferring into broth culture medium for amplification culture until effective bacteria content is 4 × 109One by mL, namely;
wherein, the trichoderma harzianum liquid is specifically that the strain of trichoderma harzianum is inoculated on the slant of PDA culture medium for activation culture, then is transferred into PDA liquid culture medium for liquid culture, and then is transferred into broth culture medium for amplification culture until the effective bacterium content is 1 × 109One by mL, namely;
wherein the Bacillus mucilaginosus is prepared by placing strain of Bacillus mucilaginosus in Na2SeO30.1g, 5g of peptone, 3g of yeast extract and KH2PO41.0g、MgSO4Culturing in 1.5g, wort 150mL, and distilled water 1000mL culture medium for 3d to obtain effective bacteria content of 2 × 108One by mL, namely; the bacillus thuringiensis liquid is prepared by inoculating bacillus thuringiensis strain to broth agar culture medium slant for slant activation training, culturing for 10 days, and transferring to culture medium composed of diammonium citrate 2.5g, peptone 30g, yeast extract 0.5g, and CH3CH2COONa 2.5g、KH2PO41.0g、MgSO4Culturing in a culture medium composed of 0.8g, wort 100mL, and distilled water 1000mL for 3d to obtain a culture medium with effective bacteria content of 3 × 108The dosage is one/mL.
Example 2:
an improved sugarcane cross breeding method comprises the following specific steps:
(1) parent breeding: the method comprises the following steps of (1) sowing the easy-to-flower parent in spring of the year, sowing the difficult-to-flower parent in advance compared with the easy-to-flower parent, and stopping fertilizing in the last 6 months of each year;
(2) marking parents: making a corresponding parent bar code label for each parent to distinguish and mark, and suspending the label at a parent planting place in the last 10 months each year, wherein the parent bar code label comprises the following information: parent name, bar code corresponding to the parent name, parent source, flowering sequence, past paternity and pollen fertility;
(3) collecting flower spikes: when the parents have the scions 1/2 and a small amount of spikelets on the tops of the parents blossom, the parents can be collected, the parents are cut off from the base parts of the sugarcane stalks, the longest state of the sugarcane stalks is kept, most leaves are cut off, a handheld scanning printer is used for scanning a suspended parent bar code label plate, 2 new bar codes with the same name as that in the step (2) are printed, the 1 st part of the bar codes is pasted on the cut parents, 8 flowering stalks of the parents are collected at the same time, the corresponding 2 nd part of the bar codes are attached to the flowering stalks, the sugarcane stalks cut off from the base parts are transported back to a hybridization greenhouse, the flower spikes are sprayed with clear water and are inserted into a clear water stem-nourishing pool for nursing in sequence;
the stem cultivating pool is divided into two rows, each row is divided into a plurality of small cells, and a position number bar code clamping groove area is arranged on the outer side of each small cell;
scanning the parent bar code and the position bar code of the position of the parent by using a scanner, processing the parent bar code and the position bar code by using a processor, and displaying the counted and collected spica number and position in a display;
(4) and (3) detecting pollen fertility: randomly selecting 2 florescent spikelets close to the florescent spikelets from the flowering spikelets retrieved in the step (3), wherein each spikelet contains 3 anthers, selecting 3 effective florets in total, obtaining 18 anthers in total, placing the anthers on a glass slide, adding an I-KI solution for dyeing, mashing the anthers by using a glass rod, detecting the pollen fertility under a microscope, and determining the paternal and maternal characteristics of the anthers;
(5) preparing and combining: according to the collected spica and the hybridization plan, a configurable hybridization combination in the same day is made, and a combination label is printed, wherein the combination label comprises the following data: female parent name, female parent position, male parent name, male parent position, hybridization cage number, preparation date and combination number, and then placing the female parent and the male parent in the combination in the same hybridization cage; firstly fixing the position of the male parent, controlling the height of the male parent to be 50cm lower than the top end of the hybridization cage, fixing a stem-cultivating solution bottle with the volume of 10L at the corresponding position of the hybridization cage, cutting off part of internode tissues at an old incision when the lower end of the male parent is collected, and quickly inserting the cut part into the stem-cultivating solution bottle containing 5L and 150ppm of sulfurous acid solution; placing the female parent in the middle of a hybridization cage, placing the female parent below and placing the male parent above, overlapping 1/3 male and female parents, fixing a stem-nourishing solution bottle with the volume of 10L at the corresponding position of the hybridization cage, quickly cutting off partial internode tissues at an old incision when the lower end of the female parent is collected by a sharp knife, quickly inserting the internode tissues into the stem-nourishing solution bottle containing 5L and 150ppm of sulfurous acid solution, and hanging the combined label plate at the middle part of the sugarcane stem of the female parent;
(6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, 4 nodes are selected from the stem section which is 250cm below the base of the spica, the leaf sheath is removed, and the stem section is wrapped by a nutrient medium, and the rapid root promoting treatment is carried out for 15 days;
(7) artificial pollination: artificial pollination is carried out every 1 hour when 8-10 am, and pollen is floated on the female parent spica only by shaking the male parent spica; in the pollination period, if the flowering period of the male parent is ended in advance, the male parent is supplemented in time until the flowering of the female parent is basically ended, and the temperature in a hybridization greenhouse is controlled to be 28 ℃ and the relative humidity is 85 percent in the artificial pollination period;
(8) after-ripening of seeds: discarding the ear stems of the male parents after hybridization, transferring the female parents into an after-ripening chamber, checking the effect of root promotion treatment, sheathing the whole flower ears downwards by a yarn bag, putting the combined label plate into the yarn bag, and fastening the lower end to prevent the label plate from falling off;
the method for checking the root promoting treatment effect comprises the following steps:
if more fibrous roots are generated, cutting off the fibrous roots from the position 3cm below the phimosis, removing the wrapped film, and putting the phimosis into flowing clear water for culturing until the seeds are mature;
if no fibrous root is produced, rapidly cutting the internode tissue of 10cm at the lower end of the female parent by using a sharp knife, rapidly inserting the internode tissue into a barrel containing 20L of a sulfurous acid solution of 150ppm to produce more fibrous roots, cutting off the internode tissue 3cm below the phimosis, removing a wrapped film, and putting the internode tissue into flowing clear water for culturing until the seeds are mature;
the gauze bag is made of transparent gauze materials, one end of the gauze bag is sealed, the other end of the gauze bag is open, the diameter of the gauze bag is 40cm, the length of the gauze bag is 80cm, and the open end of the gauze bag is provided with a lock catch.
(9) Harvesting seeds: when the seeds at the top of the female parent are white and flocculent and the small spike falls off by more than two thirds, cutting off the flower spike, putting the flower spike into a drying room for drying treatment, then removing a yarn bag, putting the seeds into a waterproof paper bag, and storing the seeds for a long time at the temperature of-15 ℃;
the conditions of the drying treatment are as follows: drying at 11 deg.C and 13% humidity for 4 days;
the nutrient medium is prepared from the following raw materials in parts by weight, wherein the nutrient medium contains 58% of organic matters, 3% of ammonium nitrogen, 0.1% of nitrate nitrogen, 20% of total nutrients, 8% of available phosphorus and 10.9% of available potassium: 0.5 part of indolebutyric acid, 0.5 part of naphthylacetic acid, 1.6 parts of rooting powder, 5 parts of alcohol and 0.7 part of compound amino acid injection, wherein ceramsite is subjected to ultramicro grinding, then the ceramsite is added into a mortar, olive oil is added into the mortar, the mixture is ground for 30 seconds, and then 25 parts of modified ceramsite powder, 20 parts of coconut coir and 10 parts of compound microbial fertilizer are obtained by irradiating 8 seconds with gamma rays.
The compound microbial fertilizer is prepared by mixing the following raw materials in percentage by weight: 3% of bacillus licheniformis liquid, 3% of bacillus megaterium liquid, 2% of yellow mould liquid, 3% of jelly sample bacillus liquid, 3% of bacillus thuringiensis liquid and 3% of trichoderma harzianum liquid, and the balance being base materials which are supplemented to 100%, wherein the base materials comprise peat, ground phosphate rock and waste sugar residues according to the weight ratio of 4:2: 3;
wherein the Bacillus licheniformis solution is prepared by placing strain of Bacillus licheniformis in Na2SeO30.1g、CH3COONa5g、NaHCO38g、(NH4)2SO48g、NH4Cl 3g、K2HPO44g、KH2PO40.5g、NaCl 0.6g、MgSO40.3g、CaCl20.2g of yeast extract, 15g of peptone and 1L of distilled water, and culturing in a culture medium prepared by adjusting the pH value to 7.3 with acetic acid to obtain the bacillus licheniformis with the effective bacteria content of 5 × 108One by mL, namely;
wherein the Bacillus megaterium solution is prepared by placing Bacillus megaterium strain in Na2SeO30.2g、CH3CH2COONa2.5g、(NH4)2SO41.5g、K2HPO40.8g、KH2PO40.3g、MgSO40.25g、NaCl 0.15g、CaCl20.3g of yeast extract, 2.0g of yeast extract and 1000mL of distilled water are cultured for 5d to obtain the culture medium with the effective bacteria content of 3 × 109One by mL, namely;
wherein the yellow mould liquid is prepared by inoculating yellow mould strain onto PDA culture medium slant for activation culture, transferring into PDA liquid culture medium for liquid culture, and transferring into broth culture mediumPerforming amplification culture until the effective bacteria content is 6 × 109One by mL, namely;
wherein, the trichoderma harzianum liquid is specifically that the strain of trichoderma harzianum is inoculated on the slant of PDA culture medium for activation culture, then is transferred into PDA liquid culture medium for liquid culture, and then is transferred into broth culture medium for amplification culture until the effective bacterium content is 3 × 109One by mL, namely;
wherein the Bacillus mucilaginosus is prepared by placing strain of Bacillus mucilaginosus in Na2SeO30.15g, 10g of peptone, 5g of yeast extract and KH2PO42.0g、MgSO42.5g, 150mL of wort and 1000mL of distilled water, and culturing for 5d to obtain a culture medium with an effective bacteria content of 6 × 108One by mL, namely; the bacillus thuringiensis liquid is prepared by inoculating bacillus thuringiensis strain to broth agar culture medium slant for slant activation training, culturing for 20 days, and transferring to culture medium composed of diammonium citrate 2.8g, peptone 30g, yeast extract 1.0g, and CH3CH2COONa3.5g、KH2PO42.0g、MgSO4Culturing in 1.2g culture medium containing wort 100mL and distilled water 1000mL for 5d to obtain effective bacteria content of 5 × 108The dosage is one/mL.
Example 3:
an improved sugarcane cross breeding method comprises the following specific steps:
(1) parent breeding: the method comprises the following steps of (1) sowing the easy-to-flower parent in spring of the year, sowing the difficult-to-flower parent in advance compared with the easy-to-flower parent, and stopping fertilizing in the last 6 months of each year;
(2) marking parents: making a corresponding parent bar code label for each parent to distinguish and mark, and suspending the label at a parent planting place in the last 10 months each year, wherein the parent bar code label comprises the following information: parent name, bar code corresponding to the parent name, parent source, flowering sequence, past paternity and pollen fertility;
(3) collecting flower spikes: when the parents have the scions 1/3 and a small amount of spikelets on the tops of the parents blossom, the parents can be collected, the parents are cut off from the base parts of the sugarcane stalks, the longest state of the sugarcane stalks is kept, most leaves are cut off, a handheld scanning printer is used for scanning a suspended parent bar code label plate, 2 new bar codes with the same name as that in the step (2) are printed, the 1 st part of the bar codes is pasted on the cut parents, 5 flowering rachis of the parents are collected at the same time, the parents are collected in a storage box and attached with the corresponding 2 nd part of the bar codes, the sugarcane stalks cut off from the base parts are transported back to a hybridization greenhouse, the flower scions are sprayed with clear water and are inserted into a clear water stem;
the stem cultivating pool is divided into two rows, each row is divided into a plurality of small cells, and a position number bar code clamping groove area is arranged on the outer side of each small cell;
scanning the parent bar code and the position bar code of the position of the parent by using a scanner, processing the parent bar code and the position bar code by using a processor, and displaying the counted and collected spica number and position in a display;
(4) and (3) detecting pollen fertility: randomly selecting 2 florescent spikelets close to the florescent spikelets from the flowering spikelets retrieved in the step (3), wherein each spikelet contains 3 anthers, selecting 3 effective florets in total, obtaining 18 anthers in total, placing the anthers on a glass slide, adding an I-KI solution for dyeing, mashing the anthers by using a glass rod, detecting the pollen fertility under a microscope, and determining the paternal and maternal characteristics of the anthers;
(5) preparing and combining: according to the collected spica and the hybridization plan, a configurable hybridization combination in the same day is made, and a combination label is printed, wherein the combination label comprises the following data: female parent name, female parent position, male parent name, male parent position, hybridization cage number, preparation date and combination number, and then placing the female parent and the male parent in the combination in the same hybridization cage; firstly fixing the position of the male parent, controlling the height of the male parent to be 40cm lower than the top end of the hybridization cage, fixing a stem-cultivating solution bottle with the volume of 7L at the corresponding position of the hybridization cage, cutting off part of internode tissues at an old incision when the lower end of the male parent is collected, and quickly inserting the cut part into the stem-cultivating solution bottle containing 5L and 150ppm of sulfurous acid solution; placing the female parent in the middle of a hybridization cage, placing the female parent below and placing the male parent above, overlapping 1/3 male and female parents, fixing a stem-nourishing solution bottle with the volume of 8L at the corresponding position of the hybridization cage, quickly cutting off part of internode tissues at an old incision when the lower end of the female parent is collected by a sharp knife, quickly inserting the internode tissues into the stem-nourishing solution bottle containing 5L and 150ppm of sulfurous acid solution, and hanging the combined label plate at the middle part of the stem of the female parent;
(6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, 3 nodes are selected from a stem section 230cm below the base of the spica, the leaf sheath is removed, the stem section is wrapped by a nutrient medium, and the rapid root promoting treatment is carried out for 10 days;
(7) artificial pollination: artificial pollination is carried out every 1 hour when 8-10 am, and pollen is floated on the female parent spica only by shaking the male parent spica; in the pollination period, if the flowering period of the male parent is ended in advance, the male parent is supplemented in time until the flowering of the female parent is basically ended, and the temperature in a hybridization greenhouse is controlled to be 26 ℃ and the relative humidity is 80% in the artificial pollination period;
(8) after-ripening of seeds: discarding the ear stems of the male parents after hybridization, transferring the female parents into an after-ripening chamber, checking the effect of root promotion treatment, sheathing the whole flower ears downwards by a yarn bag, putting the combined label plate into the yarn bag, and fastening the lower end to prevent the label plate from falling off;
the method for checking the root promoting treatment effect comprises the following steps:
if more fibrous roots are generated, cutting off the fibrous roots from 2.5cm below the phimosis, removing the wrapped film, and putting the phimosis into flowing clear water for culturing until the seeds are mature;
if no fibrous root can be generated, cutting the internode tissue of 8cm at the lower end of the female parent by a sharp knife, quickly inserting the internode tissue into a barrel containing 20L of sulfurous acid solution of 150ppm, cutting off the internode tissue from 2.5cm below the phimosis after more fibrous roots are generated, removing a wrapped film, and putting the internode tissue into flowing clear water for culturing until the seeds are mature;
the gauze bag is made of transparent gauze materials, one end of the gauze bag is sealed, the other end of the gauze bag is open, the diameter of the gauze bag is 40cm, the length of the gauze bag is 100cm, and the open end of the gauze bag is provided with a lock catch.
(9) Harvesting seeds: when the seeds at the top of the female parent are white and flocculent and the small spike falls off by more than two thirds, cutting off the flower spike, putting the flower spike into a drying room for drying treatment, then removing a yarn bag, putting the seeds into a waterproof paper bag, and storing the seeds for a long time at the temperature of minus 18 ℃;
the conditions of the drying treatment are as follows: drying at 11 deg.C and 13% humidity for 4 days;
the nutrient medium is prepared from the following raw materials in parts by weight, wherein the nutrient medium contains 63% of organic matters, 3.6% of ammonium nitrogen, 0.4% of nitrate nitrogen, 18% of total nutrients, 7% of available phosphorus and 8% of available potassium: 0.3 part of indolebutyric acid, 0.4 part of naphthylacetic acid, 0.9 part of rooting powder, 4 parts of alcohol and 0.6 part of compound amino acid injection, wherein ceramsite is subjected to ultramicro grinding, then the ceramsite is added into a mortar, olive oil is added into the mortar, the mixture is ground for 25 seconds, and then 17 parts of modified ceramsite powder, 15 parts of coconut coir and 8 parts of compound microbial fertilizer are obtained by irradiating 6 seconds with gamma rays.
The compound microbial fertilizer is prepared by mixing the following raw materials in percentage by weight: 2% of bacillus licheniformis liquid, 3% of bacillus megaterium liquid, 1% of yellow mold liquid, 2% of bacillus mucilaginosus liquid, 2% of bacillus thuringiensis liquid, 2% of trichoderma harzianum liquid, and the balance being base material supplemented to 100%, wherein the base material comprises peat, ground phosphate rock and waste sugar residue according to the weight ratio of 3:2: 2;
wherein the Bacillus licheniformis solution is prepared by placing strain of Bacillus licheniformis in Na2SeO30.09g、CH3COONa4g、NaHCO36g、(NH4)2SO47g、NH4Cl 2.5g、K2HPO43.5g、KH2PO40.3g、NaCl 0.55g、MgSO40.2g、CaCl20.2g of yeast extract, 15g of peptone and 1L of distilled water, and culturing in a culture medium prepared by adjusting the pH value to 7.3 with acetic acid to obtain the bacillus licheniformis with the effective bacteria content of 4 × 108One by mL, namely;
wherein the Bacillus megaterium solution is prepared by placing Bacillus megaterium strain in Na2SeO30.15g、CH3CH2COONa2.0g、(NH4)2SO41.7g、K2HPO40.7g、KH2PO40.2g、MgSO40.22g、NaCl 0.12g、CaCl20.24g of yeast extract, 1.7g of yeast extract and 1000mL of distilled water are cultured for 4d to obtain the effective bacteria content of 2.5 × 109One by mL, namely;
wherein the yellow mold liquid is obtained by inoculating yellow mold strain into PDA culture mediumPerforming activation culture on the above powder, transferring into PDA liquid culture medium for liquid culture, transferring into broth culture medium for amplification culture to effective bacteria content of 5 × 109One by mL, namely;
wherein, the trichoderma harzianum liquid is specifically that the strain of trichoderma harzianum is inoculated on the slant of PDA culture medium for activation culture, then is transferred into PDA liquid culture medium for liquid culture, and then is transferred into broth culture medium for amplification culture until the effective bacterium content is 2 × 109One by mL, namely;
wherein the Bacillus mucilaginosus is prepared by placing strain of Bacillus mucilaginosus in Na2SeO30.12g, peptone 7g, yeast extract 4g, KH2PO41.5g、MgSO4Culturing in 1.8g, wort 150mL, and distilled water 1000mL culture medium for 4d to obtain effective bacteria content of 3 × 108One by mL, namely; the bacillus thuringiensis liquid is prepared by inoculating bacillus thuringiensis strain to broth agar culture medium slant for slant activation training, culturing for 15 days, and transferring to culture medium composed of diammonium citrate 2.6g, peptone 30g, yeast extract 0.8g, CH3CH2COONa 3.0g、KH2PO41.5g、MgSO4Culturing 4d in 1.0g, wort 100mL, and distilled water 1000mL to obtain culture medium with effective bacteria content of 4 × 108The dosage is one/mL.
Example 4:
the procedure and the specific operation are basically the same as those in example 3, except that: the nutrient medium is formed by mixing rooting powder, coconut husk and compound fertilizer.
Example 5:
the procedure and the specific operation are basically the same as those in example 3, except that: the nutrient medium does not contain compound amino acid injection.
Example 6:
the procedure and the specific operation are basically the same as those in example 3, except that: the nutrient medium does not contain modified ceramsite powder.
Example 7:
the procedure and the specific operation are basically the same as those in example 3, except that: the nutrient medium does not contain compound microbial fertilizer.
Example 8:
the procedure and the specific operation are basically the same as those in example 3, except that: the compound microbial fertilizer in the nutrient medium does not contain basic materials.
The method of the embodiment 1 to 8 is adopted to wrap the female parent with the nutrient medium to promote the root, the condition of the root hair is observed after the treatment is finished, and 10 plants are randomly selected to record the mean value of the following items:
detecting items Number of root hairs/strip Root length/cm Root diameter/mm
Example 1 72 8.80 1.2
Example 2 92 8.68 1.2
Example 3 65 10.55 1.3
Example 4 43 3.15 0.5
Example 5 38 4.21 0.8
Example 6 41 4.14 1.0
Example 7 45 3.56 0.8
Example 8 39 3.58 0.9
From the above table, it can be seen that after comprehensive utilization of the nutrient medium of the present application for phimosis treatment, the root hairs have excellent performance in number, length and diameter.
The method comprises the steps of selecting 20 plants in each example from female parents subjected to stem wrapping in examples 1-8, carrying out clear water breeding on the female parents under the same other conditions, observing the survival condition of the female parents after 40 days of breeding, wherein the number of the survived plants in examples 1-8 is 20, 17, 18, 19 and 19 respectively; the maternal growth conditions of examples 1-8 were: growing 3 new leaves and green sugarcane leaves, growing 1 new leaf and green sugarcane leaves, growing 2 new leaves and green sugarcane leaves, and growing 2 new leaves and green sugarcane leaves.
Comparing the method of examples 1-3 (mother culture first and then stem wrapping) with comparative examples 1 and 2, the difference of comparative example 1 is that the mother culture only carries out stem culture treatment; the difference of the comparative example 2 is that the female parent is treated in a manner of wrapping the stem and then culturing the stem;
Figure BDA0001334754050000131
Figure BDA0001334754050000141
remarking: panicle utilization rate is the effective number of phimosis/total phimosis used
As can be seen from the above table, the utilization rate of the flower spikes is far higher than that of the stem raising treatment or the stem raising mode after the stem is wrapped by adopting the stem raising mode of the invention.
Figure BDA0001334754050000142
The seed setting rate is represented by the average germination number; the higher the average germination number, the higher the seed setting rate; the number of non-germinating (germination number is 0) flower ears is reduced in 2016 than in 2015, which also indicates that the seed setting rate is improved.
The above examples are merely illustrative of several embodiments of the present invention, and the description thereof is more specific and detailed, but not to be construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the appended claims.

Claims (6)

1. An improved sugarcane cross breeding method is characterized in that: the method comprises the following specific steps:
(1) parent breeding: the method comprises the following steps of (1) sowing the easy-to-flower parent in spring of the year, sowing the difficult-to-flower parent in advance compared with the easy-to-flower parent, and stopping fertilizing in the last 6 months of each year;
(2) marking parents: making corresponding parent bar code labels for each parent to carry out distinguishing marking, and suspending the labels at parent planting places in 10 months each year;
(3) collecting flower spikes: the parents have the scions 1/3-1/2, the parents can be collected when a small amount of spikelets are bloomed at the tops, the parents are cut from the base parts of the sugarcane stalks, the longest state of the sugarcane stalks is kept, most of leaves are cut off, a handheld scanning printer is used for scanning a suspended parent bar code label plate, 2 new bar codes with the same name as that in the step (2) are printed, the 1 st part of the bar codes is pasted on the cut parents, 3-8 branches of flowering stalks of the parents are collected and collected in a storage box, the corresponding 2 nd part of the bar codes are attached, the sugarcane stalks cut at the base parts are transported back to a hybridization greenhouse, the spikes are sprayed with clear water and are inserted into a clear water stem-culturing pool for conservation in sequence;
the stem cultivating pool is divided into two rows, each row is divided into a plurality of small cells, and a position number bar code clamping groove area is arranged on the outer side of each small cell;
scanning the parent bar code and the position bar code of the position of the parent by using a scanner, processing the parent bar code and the position bar code by using a processor, and displaying the counted and collected spica number and position in a display;
(4) and (3) detecting pollen fertility: randomly selecting 2 florescent spikelets close to the florescent spikelets from the flowering spikelets retrieved in the step (3), wherein each spikelet contains 3 anthers, selecting 3 effective florets in total, obtaining 18 anthers in total, placing the anthers on a glass slide, adding an I-KI solution for dyeing, mashing the anthers by using a glass rod, detecting the pollen fertility under a microscope, and determining the paternal and maternal characteristics of the anthers;
(5) preparing and combining: according to the collected spica and the hybridization plan, making a hybridization combination which can be configured on the same day, printing a combination label plate, and placing a female parent and a male parent in the combination into the same hybridization cage; firstly fixing the position of the male parent, controlling the height of the male parent to be 30-50cm lower than the top end of the hybridization cage, fixing a stem-cultivating solution bottle with the volume of 5-10L at the corresponding position of the hybridization cage, cutting off part of internode tissues at an old incision when the lower end of the male parent is collected, and quickly inserting the cut part into the stem-cultivating solution bottle containing 5L and 150ppm of sulfurous acid solution; placing the female parent in the middle of a hybridization cage, placing the female parent below and placing the male parent above, overlapping 1/3 male and female parents, fixing a stem-nourishing solution bottle with the volume of 5-10L at the corresponding position of the hybridization cage, quickly cutting off part of internode tissues at an old incision when the lower end of the female parent is collected by a sharp knife, quickly inserting the internode tissues into the stem-nourishing solution bottle containing 5L and 150ppm of sulfurous acid solution, and hanging the combined label plate at the middle part of the sugarcane stem of the female parent;
(6) and (3) female parent root promoting treatment: after the female parent is determined and fixed, selecting 2-4 nodes on a stem section which is 200-250 cm below the base of the spica, removing leaf sheaths, wrapping with a nutrient medium, and carrying out rapid root promotion treatment for 7-15 days;
the nutrient content of the nutrient medium is calculated according to the mass percentage, and the nutrient medium contains 55 to 65 percent of organic matter, 3 to 4 percent of ammonium nitrogen, 0.1 to 0.5 percent of nitrate nitrogen, 10 to 20 percent of total nutrient, 5 to 8 percent of quick-acting phosphorus and 8 to 15 percent of quick-acting potassium;
the nutrient medium is prepared from the following raw materials in parts by weight: 0.1-0.5 part of indolebutyric acid, 0.1-0.5 part of naphthylacetic acid, 0.5-1.6 parts of rooting powder, 1-5 parts of alcohol, 0.3-0.7 part of compound amino acid injection, 10-25 parts of modified ceramsite powder, 10-20 parts of coconut coir, and 5-10 parts of compound microbial fertilizer;
carrying out superfine grinding on ceramsite by using the modified ceramsite powder, adding the modified ceramsite powder into a mortar, adding olive oil into the mortar, grinding for 20-30S, and then irradiating for 5-8S by using gamma rays;
the compound microbial fertilizer is prepared by mixing the following raw materials in percentage by weight: 1-3% of bacillus licheniformis liquid, 2-3% of bacillus megaterium liquid, 1-2% of yellow mould liquid, 1-3% of bacillus mucilaginosus liquid, 2-3% of bacillus thuringiensis liquid, 1-3% of trichoderma harzianum liquid, and the balance of base materials are supplemented to 100%;
wherein the effective bacterium content of the bacillus licheniformis liquid is 3 × 108-5×108The effective bacterium content of the bacillus megaterium liquid is 2 × 109-3×109The effective bacterium content of the yellow mold bacterium liquid is 4 × 109-6×109The effective bacterium content of the bacillus mucilaginosus liquid is 2 × 108-6×108The effective bacteria content of the bacillus thuringiensis liquid is3×108-5×108The effective bacterium content of the trichoderma harzianum liquid is 1 × 10 per mL9-3×109Per mL;
the base material comprises peat, ground phosphate rock and waste sugar residues according to the weight ratio of 2-4:2: 2-3;
(7) artificial pollination: artificial pollination is carried out every 1 hour when 8-10 am, and only the male parent flower spikes are knocked to ensure that pollen falls onto the female parent flower spikes; during pollination, if the flowering period of the male parent is ended in advance, the male parent is supplemented in time until the flowering of the female parent is basically ended, and the temperature in a hybridization greenhouse is controlled to be 25-28 ℃ and the relative humidity is controlled to be 75-85% during artificial pollination;
(8) after-ripening of seeds: discarding the ear stems of the male parents after hybridization, transferring the female parents into an after-ripening chamber, checking the effect of root promotion treatment, sheathing the whole flower ears downwards by a yarn bag, putting the combined label plate into the yarn bag, and fastening the lower end to prevent the label plate from falling off;
the method for checking the root promoting treatment effect comprises the following steps:
if more fibrous roots are generated, cutting off the fibrous roots 2-3cm below the phimosis, removing the wrapped film, and putting the phimosis into flowing clear water for culturing until the seeds are mature;
if no fibrous root can be produced, cutting off the internode tissue of 5-10cm lower end of the female parent with a sharp knife, quickly inserting into a barrel containing 20L and 150ppm of sulfurous acid solution, cutting off 2-3cm below phimosis after more fibrous root is produced, removing the wrapped film, and putting into flowing clear water for culturing until the seed is mature;
(9) harvesting seeds: when the seeds at the top of the female parent are white and flocculent and the spikelets fall off by more than two thirds, cutting off the spikelets containing the yarn bags, putting the spikelets into a drying room for drying treatment, then removing the yarn bags, putting the seeds into waterproof paper bags, and storing the waterproof paper bags for a long time at the temperature of minus 15 to minus 21 ℃;
the conditions of the drying treatment are as follows: the temperature is 11 ℃, the humidity is 13%, and the drying is carried out for 4 days.
2. An improved method of cross-breeding sugar cane according to claim 1, wherein: the bacillus licheniformis liquid is prepared by placing the strain of bacillus licheniformisFrom Na2SeO30.08-0.1g、CH3COONa 3-5g、NaHCO35-8g、（NH4）2SO46-8g、NH4Cl 2-3g、K2HPO43-4g、KH2PO40.2-0.5g、NaCl 0.5-0.6g、MgSO40.1-0.3g、CaCl20.2g of yeast extract, 15g of peptone 5-10g and 1L of distilled water, and culturing in a culture medium with pH adjusted to 7.3 with acetic acid to obtain Bacillus licheniformis with effective bacteria content of 3 × 108-5×108One by mL, namely;
the bacillus megaterium liquid is prepared by placing the strain of bacillus megaterium in Na2SeO30.1-0.2g、CH3CH2COONa1.5-2.5g、(NH4)2SO41.0-1.5g、K2HPO40.6-0.8g、KH2PO40.1-0.3g、MgSO40.2-0.25g、NaCl0.1-0.15g、CaCl20.2-0.3g of yeast extract, 1.5-2.0g of yeast extract and 1000mL of distilled water are prepared into a culture medium, and the culture medium is cultured for 3-5d to obtain the culture medium with the effective bacteria content of 2 × 109-3×109The dosage is one/mL.
3. The improved sugarcane cross-breeding method as claimed in claim 1, wherein the yellow mold liquid is obtained by inoculating yellow mold strains to PDA culture medium slant for activation culture, transferring to PDA liquid culture medium for liquid culture, transferring to broth culture medium for amplification culture to effective bacteria content of 4 × 109-6×109One by mL, namely;
the trichoderma harzianum liquid is characterized in that trichoderma harzianum strains are inoculated on a PDA culture medium inclined plane for activated culture, then transferred into a PDA liquid culture medium for liquid culture, and then transferred into a broth culture medium for amplification culture until the effective bacteria content is 1 × 109-3×109The dosage is one/mL.
4. An improved method of cross-breeding sugar cane according to claim 1, wherein: the Bacillus mucilaginosus is prepared by gelatinizing liquid bacillus mucilaginosusThe strain of Bacillus is laid by Na2SeO30.1-0.15g, 5-10g of peptone, 3-5g of yeast extract and KH2PO41.0-2.0g、MgSO4Culturing in 1.5-2.5g culture medium containing wort 150mL and distilled water 1000mL for 3-5d to obtain effective bacteria content of 2 × 108-6×108One by mL, namely; the bacillus thuringiensis liquid is prepared by inoculating bacillus thuringiensis strain to broth agar culture medium slant for slant activation training, culturing for 10-20 days, and transferring to culture medium composed of diammonium citrate 2.5-2.8g, peptone 30g, yeast extract 0.5-1.0g, and CH3CH2COONa 2.5-3.5g、KH2PO41.0-2.0g、MgSO4Culturing in 0.8-1.2g culture medium containing wort 100mL and distilled water 1000mL for 3-5 days to obtain culture medium with effective bacteria content of 3 × 108-5×108The dosage is one/mL.
5. An improved method of cross-breeding sugar cane according to claim 1, wherein: in step (2), the parent barcode label contains the following data: parent name, bar code corresponding to the parent name, parent source, flowering sequence, past paternity and pollen fertility.
6. An improved method of cross-breeding sugar cane according to claim 1, wherein: in step (5), the combination tag includes the following data: female parent name, female parent position, male parent name, male parent position, hybrid cage number, preparation date and combination number.
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