CN103918554A - Method for inducing tetraploid garlic by using oryzalin - Google Patents
Method for inducing tetraploid garlic by using oryzalin Download PDFInfo
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- CN103918554A CN103918554A CN201410143290.5A CN201410143290A CN103918554A CN 103918554 A CN103918554 A CN 103918554A CN 201410143290 A CN201410143290 A CN 201410143290A CN 103918554 A CN103918554 A CN 103918554A
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- oryzalin
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- tetraploid
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Abstract
The invention discloses a method for inducing tetraploid garlic by using oryzalin. According to the method, the oryzalin is selected as a mutagenic agent for inducing the tetraploid garlic, an optimal mutagenizing method is adopted, and the best effect of inducing tetraploid garlic is obtained through research, and thus germplasm resource reserve is provided for germplasm innovation and breeding of new variety of the garlic.
Description
Technical field
The invention belongs to agricultural technology field, in particular to a kind of tetraploid method of oryzalin induction garlic of utilizing.
Background technology
Colchicine is the chemical mutagen of conventional induce chromosome redoublement.Oryzalin is compared with colchicine, has less toxicity, and chromosome doubling better effects if.At present, oryzalin all successfully induces polyploid (Chalak1996 in various crop;
2000; Carvalho et al.2005;
m.M.et al., 2011), also be applied in addition potato (Leire Barandalla et al., 2006), rose (J.F.Allum et al., 2007), Hubei cercis (Joshua D.Nadlera et al., 2012), lily (letter is good etc., 2013), view and admire ginger (HamidouF., 2009), watermelon (Yan Zhihong etc., 2008; Zhao Yijie etc., 2011), on the crop such as Pon mandarin orange (He Jian etc., 2011) orchid (TildenP. etc., 2011).A < < Melon tetraploid induction method > > patent of invention (application number CN201310258679) is also the oryzalin induction tetraploid of use.
Garlic be take Clone Selection And Reproduction as main, is difficult to carry out garlic germplasm innovation and breeding of new variety.And ploidy breeding is a kind of effective way of garlic germplasm innovation and breeding of new variety.Garlic utilizes colchicine-induced polyploid successfully to report (Zhang Suzhi etc., 2006) at present, has no and utilizes oryzalin induction garlic tetraploid.Although colchicine is conventional polyploid, double mutagen, toxicity is large, and cost is high, efficiency of inducing mutation is low, yet, directly use oryzalin induction garlic tetraploid success rate low, how improving garlic tetraploid success rate is art technology technical problem urgently to be resolved hurrily.
Summary of the invention
Therefore for above shortcoming, it is induction garlic tetraploid mutagen that the present invention selects oryzalin, by rational selection mutagenic condition, obtains tetraploid plant, for garlic germplasm innovation and breeding of new variety provide germ plasm resource deposit.In order to realize object of the present invention, intend adopting following technical scheme:
One aspect of the present invention relates to a kind of tetraploid method of oryzalin induction garlic of utilizing, and it is characterized in that comprising the steps:
(1) 1.0mg6-benzyl aminoadenine, 0.5mg methyl α-naphthyl acetate are dissolved in 1LB5 minimal medium, are mixed with two kinds of medium of liquid nutrient medium and solid culture medium, two kinds of medium pHs are adjusted to 5.8-6.0, the horizontal high voltage sterilizing of going forward side by side;
(2) will after 2-12mg oryzalin filtration sterilization, join in the liquid nutrient medium that 1L autoclaving is good, packing is standby;
(3) garlic bulb is through naturally drying after breaking dormancy, its bulbil peeling is rinsed to 0.25-1.0h under running water, then under aseptic condition, use 75%(mass percentage concentration) alcohol disinfecting soak 0.51.0-2.0min, under anatomical lens, separate stem apex with 2 leaf primordium 0.15-0.25mm left and right as explant;
(4) Shoot tip explants of peeling off is seeded in to the B5 that contains 2-12mg/L oryzalin and divides in the liquid nutrient medium installing, secretly cultivate 48-144h; , then taking out stem apex and in sterile water, rinse 3-5 time, and then stem apex is proceeded to solid culture medium, condition of culture is 23.0 ± 2.0 ℃ of temperature, illumination 25～31 μ molm
-2s
-1, illumination 16h/d.
(5) stem apex is cultivated after 50-70d on solid culture medium, forms the bud seedling (plant) with root.
(6) plant of acquisition is carried out to Ploidy Identification, by two kinds of methods, identify, first with plant leaf, by flow cytometer, detect, then count evaluation by root tip chromosomes, further verify the ploidy of plant, thereby identify the tetraploid plant of acquisition.
It is induction garlic tetraploid mutagen that the present invention selects oryzalin, by research, obtains the best tetraploid effect of induction garlic, for garlic germplasm innovation and breeding of new variety provide germ plasm resource deposit.
Accompanying drawing explanation
Fig. 1: chromosome counting qualification result: left figure wherein: dliploid chromosome; Right figure: tetraploid chromosomes.
Fig. 2: flow cytometer testing result: left figure wherein: dliploid DNA content; Right figure: tetraploid DNA content.
Embodiment
Embodiment 1
Mutagenic treatment method: garlic bulb is through naturally drying after breaking dormancy, the bulbil of its peeling is rinsed to 0.5h under running water, then under aseptic condition with 75% alcohol disinfecting 1min, under anatomical lens, separate stem apex with 2 leaf primordium 0.2mm left and right as explant.Be seeded in the liquid nutrient medium of the B5+6-BA1.0mg/L+NAA0.5mg/L that contains 2-12mg/L oryzalin the dark processing time 48-144h setting that cultivates to; Then take out stem apex and in sterile water, rinse 3-5 time, then stem apex is proceeded to solid culture medium, condition of culture is 23.0 ± 2.0 ℃ of temperature, illumination 25～31 μ molm-2s
-1, 16h/d.
Tetraploid authentication method: the plant of acquisition is carried out to Ploidy Identification, detect with two kinds of methods of chromosome counting and identify by flow cytometer.Obtain 25.6% tetraploid induction rate.And do contrast with colchicine, compare, colchicine tetraploid pick-up rate is up to 18.9%.
The above, be only the specific embodiment of the present invention, but protection scope of the present invention is not limited to this, and any variation of expecting without creative work or replacement, within all should being encompassed in protection scope of the present invention.Therefore, protection scope of the present invention should be as the criterion with the protection domain that claims were limited.
Claims (2)
1. utilize the tetraploid method of oryzalin induction garlic, it is characterized in that comprising the steps:
(1) 1.0mg6-benzyl aminoadenine, 0.5mg methyl α-naphthyl acetate are dissolved in 1LB5 minimal medium, are mixed with two kinds of medium of liquid nutrient medium and solid culture medium, two kinds of medium pHs are adjusted to 5.8-6.0, the horizontal high voltage sterilizing of going forward side by side;
(2) will after 2-12mg oryzalin filtration sterilization, join in the liquid nutrient medium that 1L autoclaving is good, packing is standby;
(3) garlic bulb is through naturally drying after breaking dormancy, its bulbil peeling is rinsed to 0.5-1.0h under running water, then under aseptic condition, use 75%(mass percentage concentration) alcohol-pickled 1.0-2.0min, under anatomical lens, separate stem apex with 2 leaf primordium 0.15-0.25mm left and right as explant;
(4) stem apex of peeling off is seeded in the liquid nutrient medium minute installing that contains 2-12mg/L oryzalin, the dark 48-144h that cultivates, then takes out stem apex and in sterile water, rinses 3-5 time, then proceed to solid culture medium, condition of culture is 23.0 ± 2.0 ℃ of temperature, illumination 25～31 μ molm
-2s
-1, illumination 16h/d;
(5) stem apex is cultivated after 50-70d on solid culture medium, forms the bud seedling plant with root;
(6) the bud seedling plant of acquisition is carried out to Ploidy Identification, by following two kinds of methods, identify, first with plant leaf, by flow cytometer, detect, then count evaluation by root tip chromosomes, further verify the ploidy of plant; Thereby identify the tetraploid plant of acquisition.
2. method according to claim 1, is characterized in that stem apex after oryzalin processing to proceed on solid culture medium and cultivate, and in its solid culture medium, the concentration of agar powder is 0.6-0.8%.
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| CN201410143290.5A CN103918554B (en) | 2014-04-11 | 2014-04-11 | One utilizes oryzalin to induce the tetraploid method of garlic |
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| CN201410143290.5A CN103918554B (en) | 2014-04-11 | 2014-04-11 | One utilizes oryzalin to induce the tetraploid method of garlic |
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| CN103918554A true CN103918554A (en) | 2014-07-16 |
| CN103918554B CN103918554B (en) | 2016-04-20 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508676A (en) * | 2016-11-03 | 2017-03-22 | 广西玉林容山堂生物科技有限公司 | Purification and rejuvenation method for Yulin fragrant garlic |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101124888A (en) * | 2007-09-28 | 2008-02-20 | 南开大学 | Method for cultivating tetraploid of dendrobium officinale |
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2014
- 2014-04-11 CN CN201410143290.5A patent/CN103918554B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101124888A (en) * | 2007-09-28 | 2008-02-20 | 南开大学 | Method for cultivating tetraploid of dendrobium officinale |
Non-Patent Citations (4)
| Title |
|---|
| 张素芝等: "秋水仙素诱导大蒜四倍体的研究", 《核农学报》, vol. 20, no. 4, 31 December 2006 (2006-12-31), pages 303 - 308 * |
| 张雨等: "甲基胺草磷诱导大蒜多倍体的研究", 《中国蔬菜》, no. 14, 31 December 2012 (2012-12-31), pages 90 - 94 * |
| 董伟清等: "玉林大蒜茎尖脱毒与繁殖技术研究", 《广西农业科学》, vol. 40, no. 10, 31 December 2009 (2009-12-31), pages 1289 - 1291 * |
| 邵果园等: "氨磺乐灵在园艺植物倍性育种上的应用", 《中国南方果树》, vol. 42, no. 6, 31 December 2013 (2013-12-31), pages 24 - 27 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106508676A (en) * | 2016-11-03 | 2017-03-22 | 广西玉林容山堂生物科技有限公司 | Purification and rejuvenation method for Yulin fragrant garlic |
| CN106508676B (en) * | 2016-11-03 | 2018-11-06 | 广西玉林容山堂生物科技有限公司 | A kind of purification and rejuvenation method of Yulin perfume (or spice) garlic |
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| CN103918554B (en) | 2016-04-20 |
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