CN101595841B - Induction method of poplar tetraploid plants - Google Patents
Induction method of poplar tetraploid plants Download PDFInfo
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- CN101595841B CN101595841B CN2009100867735A CN200910086773A CN101595841B CN 101595841 B CN101595841 B CN 101595841B CN 2009100867735 A CN2009100867735 A CN 2009100867735A CN 200910086773 A CN200910086773 A CN 200910086773A CN 101595841 B CN101595841 B CN 101595841B
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Abstract
The invention discloses an induction method of poplar tetraploid plants, belonging to the field of plant inheritance breeding. The method of the invention comprises the following steps: pollination is carried out on female inflorescence of poplar female parent by powder of poplar male parent to form infructescence, when flocculent fiber is formed and wraps ovule in the infructescence ovary, physicochemical treatment is carried out on the infructescence, seed selection is carried out on tetraploid plants; the inventor of the invention discovers by tests that the effective period for carrying out physicochemical treatment to induce the zygote chromosome to be reduplicated is when most zygotes of the poplar is at the first mitosis process and the flocculent fiber starts to wrap but does not completely covet the ovule, thus the induction efficiency of tetraploid plants is improved. The invention avoids verbose links such as fixing, dehydrating, waxing, embedding, slicing, pigmentation, microexamination of the ovule and the like, thus enhancing the timeliness of physicochemical treatment, saving induction reagent and experimental material and improving the precision and stability of the reduplication of zygote chromosome.
Description
Technical field
The present invention relates to the abductive approach of a plant species tetraploid plant, relate in particular to a kind of abductive approach of inducing poplar tetraploid plant, belong to the plant genetics and breeding field.
Background technology
Utilizing tetraploid plant and diploid hybrid is to obtain the most direct valid approach of populus tremula plant.Some area in the Northern Europe and the U.S., even a tetraploid aspen female plant of breeding directly is planted in open pollination in the good aspen standing forest, on tetraploid aspen female plant, collect seed every year, grow seedlings (Xu Weiying. willow. Harbin: the Heilungkiang People's Press, 1988).
The poplar tetraploid seed selection can take to utilize method (the Nilsson-Ehle H.Production of forest trees with increased chromosome number and increased timberyield.Svensk Papp.Tidn. of triploid and diploid hybrid, 1938,2:5; Bergstrom I.On the progeny of diploid * triploid Populus tremula with special reference to the occurrence of tetraploid.Hereditas, 1940,26:191-201; Winton L, Einspahr DW.Tetraploid aspen productionusing unreduced triploid pollen.Forest Science, 1970,16 (2): 180-182; Harder ML, Verhagen S, Winton L, Einspahr DW.Tetraploid Aspen production using unreducedpollen from triploid males.Forest Science, 1976,22 (3): induce somatic double (Mattila RE.On theproduction of the tetraploid hybrid Aspen by colchicine treatment.Hereditas such as seed, bud, rataria or zygote 329-330) or by processing such as colchicins, 1961,47 (3/4): 631-640; Einspahr DW.Colchicine treatment of newly formed embryos ofquaking aspen.Forest Science, 1965,11 (4): 456-459; Pesina K.Experimentalinduction of polyploidy in poplars.Preslia, 1963,35 (2): 101-109; Ganchev P.Geneticinvestigation of some Aspen and hybrid (the forms obtainedby treating seeds with γ-rays of Populus alba * P.tremula) (
60Co) and colchincine solution.GorskostopanskaNauka, 1976,13 (1): 3-15; Bakulin VT.Producing polyploid forms of some poplarspecies by means of colchicine treatment.Izv.Sib.Otd.AN SSSR, ser.biol.n., 1977,2:68-75; Mashkina OS, Burdaeva LM, V ' yunova LN.Experimental mutagenesis andpolyploidy in breeding forest trees.Lesnaya Genetika, 1989,136-137; Li Yun. white poplar oogamete and somatic double induced triploid and tetraploid Study on Technology. Beijing: Beijing Forestry University, 1997).Wherein, the prerequisite of triploid and diploid hybrid is at first will obtain triploid; And utilize materials such as processing germination seeds such as colchicin or bud to form mixoplod or chimera easily.
Zygote forms the tetraploid of isozygotying, and is difficult for forming mixoplod or chimera plant as a single cell after the zygote chromosome doubling, be the most desirable selection of artificial induction's tetraploid plant.So far, have only (Mashkina OS such as Mashkina, Burdaeva LM, V ' yunova LN.Experimental mutagenesis andpolyploidy in breeding forest trees.Lesnaya Genetika, 1989,136-137) clearly report utilizes the zygote of colchicine solution Processing Ag white poplar first division process to obtain tetraploid plant.But the unresolved zygote chromosome doubling of correlative study is effectively handled the differentiation problem in period, but adopt lot of materials is handled day by day, and the big and poor repeatability of workload, and induce efficient very low.As seen, be the tetraploid key issue of the restriction efficient inducing poplar of zygote chromosome doubling first time mitosis period of how differentiating willow after fertilization zygote apace, and the solution of relevant issues is significant for poplar tetraploid inductive technology and the efficient seed selection of triploid.
Summary of the invention
Technical problem to be solved by this invention is to overcome the existing existing deficiency of poplar tetraploid abductive approach, a kind of abductive approach of new poplar tetraploid plants is provided, this method has solved willow zygote chromosome doubling and has effectively handled the differentiation problem in period, need not material is handled day by day, can effectively increase physics and chemistry and handle the accuracy and the stability of inducing the zygote chromosome doubling, improve tetraploid induction efficient, saved and induced reagent and experiment material.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of abductive approach of poplar tetraploid plants comprises: the pollen with the willow male parent is pollinated to the female inflorescence of willow female parent; Form infructescence; Infructescence is applied physics and chemistry to be handled; The seed selection tetraploid plant, promptly; Wherein, infructescence being applied the period that physics and chemistry handles is: when wadding fibrous in the infructescence ovary formed and begun to wrap up ovule infructescence is applied physics and chemistry and handle.
Better induce effect in order to reach, preferred, when beginning to wrap up the ovule base portion but still infructescence is applied colchicin during the whole cell of underfill, wadding fibrous in the infructescence ovary handle; Described colchicin is handled and is preferably that to adopt weight percent concentration be that 0.5% colchicine solution soaked infructescence 24 hours; Or when the wadding fibrous parcel ovule base portion in the infructescence ovary and middle part but still infructescence is applied high temperature treatment during the whole cell of underfill; Described high temperature treatment is preferably the Temperature Treatment infructescence 4 hours that adopts 38-44 ℃.
To which kind of situation or form, those skilled in the art can determine by perusal as for the inner wadding fibrous growth of definite ovary, also can observe and confirm that these modes are those skilled in the art institute on top of by instruments such as magnifying glasses.
Described willow female parent is preferably populus simonii (Populus simonii), false simon poplar bark (P.pseudo-simonii), Populus adenopoda (P. adenlpodn), Mao Xinyang, and (P.tomentosa * P.balleana) or wise man draw No. 3 poplars (P.pseudo-simonii * P.nigra var.lica) etc., and more preferably the wise man draws poplar No. 3; Described male parent is preferably false simon poplar bark, populus simonii, Populus adenopoda, Xinjiang poplar (P.balleana) or Beijing poplar, and (P. * beijingensis) etc. is more preferably Beijing poplar.
It can be the various tetraploid processing means of inducing that described physics and chemistry is handled, and for example, can be to handle with colchicine solution, also can be to adopt high temperature that infructescence is induced processing, and these technological means are all understood thoroughly by those skilled in the art.
The inventor finally finds in conjunction with technological means such as paraffin sections, when wadding fibrous in the willow infructescence ovary formed and begun to wrap up ovule base portion (or parcel base portion and middle part) but do not covered ovule fully as yet, the interior most zygotes of infructescence this moment enter the process of mitosis for the first time, handle the optimization process period of inducing zygote chromosome doubling for applying physics and chemistry this moment, strengthen applying the ageing of physics and chemistry processing, can effectively improve tetraploid induction efficient.The inventive method need not to experience loaded down with trivial details links such as fixing, dehydration to ovule, waxdip, embedding, section, dyeing, microexamination, also can effectively increase physics and chemistry and handle the accuracy and the stability of inducing the zygote chromosome doubling, reagent and experiment material are induced in saving, reduce production costs.
Description of drawings
Wadding fibrous parcel ovule base portion in Fig. 1 willow ovary but do not cover the form of ovule as yet fully.
Embodiment
Further describe the present invention below in conjunction with specific embodiment, advantage of the present invention and characteristics will be more clear along with description.But these embodiment only are exemplary, scope of the present invention are not constituted any restriction.It will be understood by those skilled in the art that and down can make amendment or replace without departing from the spirit and scope of the present invention, but these modifications and replacing all fall within the scope of protection of the present invention the details of technical solution of the present invention and form.
Embodiment 1
Drawing No. 3 poplar * Beijing poplars with the wise man is example, to the infructescence of the back regular period of pollinating with FAA (V (70% ethanol): V (acetate): V (38% formaldehyde)=90: 5: 5) fix, and the inner wadding fibrous developmental condition of observation ovary, by routine paraffin wax sections observation zygote developmental process, because there is certain asynchronism in its growth, zygote growth in a large amount of ovules is added up, the extinction test error, add up 213 of ovules altogether, show that the wise man draws No. 3 interior fibroflocculent generations of poplar ovary, grow and have substantial connection (table 1) with its double fertilization and zygote developmental process.Wadding fibrously in ovary begin to wrap up ovule base portion (Fig. 1), or further begin to wrap up the ovule middle part but when not covering ovule fully as yet, at this moment, most zygotes enter the first mitosis process.
With wadding fibrous developmental condition in the ovary is reference, when wadding fibrous in the ovary begins to wrap up ovule base portion (this moment for pollination after the 7th day) infructescence is applied colchicin and handle when pollination back wise man draws in No. 3 poplar * Beijing poplar infructescence, concrete processing method is: adopting weight percent concentration is 0.5% colchicine solution immersion infructescence 24 hours; Infructescence is not carried out high temperature treatment when (this moment for pollination after the 8th day) when the middle part of the wadding fibrous further parcel ovule in the ovary but when covering ovule fully as yet, concrete high-temperature processing method is for adopting 38-44 ℃ Temperature Treatment infructescence 4 hours; Result: in above-mentioned period infructescence is carried out physics and chemistry respectively and handle, total coinduction has obtained 8 strain poplar tetraploid plants.
When wadding fibrous in the infructescence ovary covers the strain handle fully and begins to be stretched to ovule (this moment for pollination after the 6th day); Or the most of ovule of wadding fibrous parcel in ovary, but when not covering fully as yet (be the 9th day after pollinating this moment), in above-mentioned period, infructescence is applied colchicin or high temperature treatment respectively, result: do not obtain a strain poplar tetraploid plants (table 2, table 3).
Above-mentioned result of the test explanation, when the wadding fibrous parcel ovule base portion in the ovary in the willow infructescence or further begin to wrap up the middle part of ovule but apply colchicin or high temperature treatment when not covering ovule fully as yet respectively, can significantly increase physics and chemistry and handle the accuracy and the stability of inducing the zygote chromosome doubling, improve the tetraploid efficient of inducing.
Table 1 wise man draws poplar * Beijing poplar zygotic embryo progenesis process No. 3
Annotate: bracket is outward the shared percentage of each developmental stage (%); In the bracket is each developmental stage statistical magnitude.
Table 2 colchicin is induced ' wise man draws poplar No. 3 ' * ' Beijing poplar ' zygote chromosome doubling seed selection tetraploid
Table 3 high temperature induction ' wise man draws poplar No. 3 ' * ' Beijing poplar ' zygote chromosome doubling seed selection tetraploid
Illustrate: contrast all refers to handle and normal infructescence of cultivating in table 2 and the table 3, and purpose is that the possibility that spontaneous doubling and the fusion of natural unreduced gamete form the polyploidy zygote does not take place for the eliminating zygote is treated.
Claims (1)
1. the abductive approach of a poplar tetraploid plants, this method may further comprise the steps: the pollen with the willow male parent is pollinated to the female inflorescence of willow female parent, forms infructescence; Infructescence is applied physics and chemistry handle the seed selection tetraploid plant; It is characterized in that: after pollination the 7th day begins to wrap up the ovule base portion when wadding fibrous in the infructescence ovary but still infructescence applied colchicin or high temperature treatment during the whole cell of underfill; It is that to adopt weight percent concentration be that 0.5% colchicine solution soaked infructescence 24 hours that described colchicin is handled; Described high temperature treatment is to adopt the Temperature Treatment 4 hours of 38 ℃ or 44 ℃;
Or after pollination the 8th day, when the wadding fibrous parcel ovule base portion in the infructescence ovary and middle part but still infructescence is applied high temperature treatment during the whole cell of underfill; Described high temperature treatment is with 41-44 ℃ Temperature Treatment 4 hours;
Wherein, described willow male parent is Beijing poplar (P. * beijingensis); Described willow female parent is that the wise man draws No. 3 poplars (P.pseudo-simonii * P.nigra var.lica).
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Families Citing this family (8)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102524056A (en) * | 2011-12-07 | 2012-07-04 | 北京林业大学 | Method for breeding poplar tetraploid plants with colchicine |
| CN102919132B (en) * | 2012-11-29 | 2013-12-11 | 西南林业大学 | Tissue culture method for inducing populus yunnanensis dode tetraploid in combination with colchicine |
| CN105684893A (en) * | 2016-02-02 | 2016-06-22 | 北京林业大学 | Breeding method of cortex eucommiae triploids |
| CN106718865A (en) * | 2016-12-09 | 2017-05-31 | 河北省农林科学院昌黎果树研究所 | A kind of method based on inflorescence dipping induction Chinese chestnut 2n pollen |
| CN106688881B (en) * | 2016-12-16 | 2018-09-21 | 北京林业大学 | One kind detaching homozygotic method from sex mosaic plant again |
| CN106613931A (en) * | 2016-12-20 | 2017-05-10 | 河北省农林科学院昌黎果树研究所 | 2n pollen induction method for Japanese chestnuts and application of 2n pollen induction method |
| CN112056212B (en) * | 2020-09-23 | 2021-08-31 | 北京林业大学 | Method for cultivating polyploid poplar and application thereof |
| CN114747480B (en) * | 2022-03-25 | 2023-04-28 | 北京林业大学 | Method for inducing forest tetraploid in field in non-isolated manner |
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