CN111758566B - Method for inducing adventitious bud regeneration of tender green semi-compact callus of fraxinus mandshurica - Google Patents
Method for inducing adventitious bud regeneration of tender green semi-compact callus of fraxinus mandshurica Download PDFInfo
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- 206010020649 Hyperkeratosis Diseases 0.000 title claims abstract description 66
- 241000565391 Fraxinus mandshurica Species 0.000 title claims abstract description 27
- 230000001939 inductive effect Effects 0.000 title claims abstract description 14
- 230000008929 regeneration Effects 0.000 title claims abstract description 11
- 238000011069 regeneration method Methods 0.000 title claims abstract description 11
- 238000000034 method Methods 0.000 title claims abstract description 7
- 235000008744 Brassica perviridis Nutrition 0.000 title claims abstract description 4
- 241000712024 Brassica rapa var. perviridis Species 0.000 title claims abstract description 4
- 230000006698 induction Effects 0.000 claims abstract description 40
- 239000001963 growth medium Substances 0.000 claims abstract description 21
- 210000002257 embryonic structure Anatomy 0.000 claims abstract description 3
- 229930006000 Sucrose Natural products 0.000 claims description 9
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 claims description 9
- 239000005720 sucrose Substances 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 6
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 6
- 239000008272 agar Substances 0.000 claims description 6
- 230000015572 biosynthetic process Effects 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 4
- 230000004083 survival effect Effects 0.000 claims description 4
- 239000004471 Glycine Substances 0.000 claims description 3
- 235000020415 coconut juice Nutrition 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 238000005286 illumination Methods 0.000 claims description 3
- 239000002609 medium Substances 0.000 claims description 3
- 238000009825 accumulation Methods 0.000 claims description 2
- 230000001954 sterilising effect Effects 0.000 claims description 2
- 238000004659 sterilization and disinfection Methods 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 230000002349 favourable effect Effects 0.000 claims 2
- 241000196324 Embryophyta Species 0.000 claims 1
- 238000005516 engineering process Methods 0.000 abstract description 7
- 238000013459 approach Methods 0.000 abstract description 3
- 239000007787 solid Substances 0.000 abstract description 2
- 229940088597 hormone Drugs 0.000 description 15
- 239000005556 hormone Substances 0.000 description 15
- 238000011282 treatment Methods 0.000 description 9
- 230000000694 effects Effects 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 4
- 230000004069 differentiation Effects 0.000 description 3
- 230000009286 beneficial effect Effects 0.000 description 2
- 238000005406 washing Methods 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 241000258746 Asterina <sea star> Species 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000372427 Justicia aurea Species 0.000 description 1
- 230000010432 cotyledon development Effects 0.000 description 1
- 230000005059 dormancy Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000012214 genetic breeding Methods 0.000 description 1
- 230000035784 germination Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 238000004161 plant tissue culture Methods 0.000 description 1
- 230000000644 propagated effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- SUKJFIGYRHOWBL-UHFFFAOYSA-N sodium hypochlorite Chemical compound [Na+].Cl[O-] SUKJFIGYRHOWBL-UHFFFAOYSA-N 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 238000004114 suspension culture Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/001—Culture apparatus for tissue culture
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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
- A01H4/00—Plant reproduction by tissue culture techniques ; Tissue culture techniques therefor
- A01H4/008—Methods for regeneration to complete plants
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- Developmental Biology & Embryology (AREA)
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- Biotechnology (AREA)
- Cell Biology (AREA)
- Botany (AREA)
- Environmental Sciences (AREA)
- Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
Abstract
The invention provides a method for realizing adventitious bud regeneration and propagation of hypocotyls and stem segments of fraxinus mandshurica through a callus approach by inducing a key link of tender green semi-compact callus. Cutting off hypocotyls from the culture of mature embryos until cotyledons are expanded (21d), and inoculating the hypocotyls in a20 culture medium; cutting stem segments from aseptic seedlings, inoculating the stem segments into a c12 culture medium, and performing dark/light alternate culture to obtain bright green semi-compact callus, wherein the induction rates of hypocotyl and stem segment callus are 88.69% and 99.15% respectively, adventitious buds are formed at semi-compact gaps of callus blocks, and the induction rates are 10.52% and 33.33% respectively. The invention solves the bottleneck link that the fraxinus mandshurica is difficult to differentiate adventitious buds through a callus approach, and lays a solid foundation for the fraxinus mandshurica rapid propagation technology through efficient callus induction and effective adventitious bud regeneration.
Description
The technical field is as follows:
the invention belongs to the field of forest genetic breeding, plant tissue culture category and a high-efficiency propagation technology of fraxinus mandshurica
Background art:
the fraxinus mandshurica seeds have the characteristics of long growth interval period, deep dormancy, low propagation speed and the like, the excellent clone and the heterosis seeds can enter the mature period within about 20 years, the cutting grafting vegetative propagation technology is limited by factors such as seasons and the like, and the excellent offspring cannot be propagated in large quantity, so that the establishment of an efficient propagation technology is particularly important. Zhang Huijun and the like performed preliminary exploration on the culture of the germination of the water willow bud, the stem tip, the seed and other explants through induction; in 2010, Shenhai dragon and others tried to establish a propagation system according to hypocotyl, but the propagation coefficient was low and unstable; inducing fraxinus mandshurica leaves and the like in 2013 years by taking fraxinus mandshurica leaves as explants to obtain fraxinus mandshurica loose callus; the fraxinus mandshurica suspension culture system is established in 2018; the fraxinus mandshurica callus is easier to obtain by inducing explants, but the study on the callus on adventitious bud induction differentiation technology is not mature at present. The invention establishes and optimizes a high-efficiency callus induction and adventitious bud regeneration system.
The invention content is as follows:
the invention provides a method for efficient callus induction and adventitious bud regeneration, which comprises the following specific contents:
1, respectively inducing regeneration of callus and adventitious buds by four fraxinus mandshurica explants (hypocotyl, root, cotyledon and stem segments), and screening to obtain a stem segment of a fraxinus mandshurica tissue culture seedling, which is the most suitable explant for inducing the callus and the adventitious buds;
2, finally selecting an MSB5 optimal culture medium by comparing the influence of three different culture media (WPM, MS and MSB5) on the adventitious bud induction rate; the culture conditions are as follows: dark culture of explants for 14 days, and light-transferring culture (illumination intensity 80. mu. mol. x.m)-2*s-116 h/8 h in dark, 25 +/-2 ℃ in temperature and 80% -90% in humidity) for 20 days, and inducing a light green semi-compact callus and inducing cluster adventitious buds for 30 days.
3, observing and counting the survival rate and the induction rate of the callus and the adventitious bud by a plurality of combined tests of exogenously adding two hormones (6-BA and TDZ), and comprehensively obtaining: the induction rates of the hypocotyls on a20(MS +30g/L sucrose +7g/L agar +5mg/L6-BA +1mg/L TDZ) culture medium and stem sections on c12(MSB5+30g/L sucrose +7g/L agar +5mg/L6-BA +8mg/L TDZ +0.1mg/L IBA +2mg/L glycine + 5% coconut water pH are both 5.8) culture medium are 88.69% and 99.15%, respectively, and the induction rates of adventitious buds are 10.5% and 33.33%, respectively.
The invention is one of the key technologies of the rapid propagation of the fraxinus mandshurica tissue culture seedling, fills the blank of the research on the rapid propagation of the tissue culture seedling in the asexual propagation field of the fraxinus mandshurica through the high-efficiency callus induction and the effective adventitious bud regeneration, solves the bottleneck link that the fraxinus mandshurica is difficult to differentiate the adventitious bud through the callus approach, and lays a solid foundation for the rapid propagation technology of the fraxinus mandshurica through the high-efficiency callus induction and the effective adventitious bud regeneration.
Description of the drawings:
FIG. 1 hypocotyl, root and cotyledon induced callus and hypocotyl regenerated adventitious bud
FIG. 2 Stem-induced callus and its regenerated adventitious bud
The specific implementation mode is as follows:
1, the concrete steps of disinfection and inoculation of fraxinus mandshurica seeds
Peeling the testa of the vernalized Fraxinus mandshurica seeds, washing with running water for 24h, treating with 70% alcohol for 30-45s, sterilizing with 10% NaClO for 15-20min, washing with sterile water for 4 times, peeling off embryos, and inoculating into WPM culture medium containing 30g/L sucrose and 7g/L agar for culture.
2, adventitious bud induction culture medium for different explants of fraxinus mandshurica tissue culture seedlings
Selecting a sterile seedling which has germinated for 21 days and has good cotyledon development and growth, cutting off hypocotyl, root and cotyledon, respectively inoculating the hypocotyl, the root and the cotyledon in different hormone combined culture media (see table 1), cutting off a non-bud stem section between two bud points of the sterile seedling of the fraxinus mandshurica, inoculating an adventitious bud induction culture medium (see table 2), culturing an induction material in a dark place for 14 days, and then performing light culture, wherein the culture conditions are as follows: illumination intensity 80 μmol m-2*s-116 h/8 h in light, 25 +/-2 ℃ in temperature and 80% -90% in humidity, 10 different tissues are treated by each hormone, and 3 experiments are repeated. And counting the callus induction rate at 20d and the adventitious bud induction rate at 30 d.
TABLE 1 Asterina mandshurica adventitious bud Induction Medium for root and cotyledon
TABLE 2 culture medium for inducing adventitious bud in stem segment of Fraxinus mandshurica
3, hypocotyl as explant inducing callus and adventitious bud
Comparing the effect of the 6-BA and TDZ hormone combinations with different concentrations on the induction of hypocotyl callus, carrying out variance analysis, and the 22 hormone treatment results show that: the a20(MS +30g/L Sucrose +7g/L Agar +5mg/L6-BA +1mg/L TDZ) culture medium is the best hypocotyl callus induction and adventitious bud regeneration culture medium.
TABLE 3 Effect of different hormone treatments on hypocotyl callus and adventitious bud Induction
Note: lower case letters indicate significant differences at the P <0.05 level
4, using cotyledon as explant to induce callus and adventitious bud
Comparing the effect of the hormone combination of 6-BA and TDZ with different concentrations on the induction of the cotyledon callus, carrying out variance analysis, and 20 hormone treatment results show that: the induction rate of the callus of a4(MS +30g/L Sucrose +7g/L Agar +1mg/L TDZ) culture medium reaches 100%, but no adventitious bud is induced by all the culture medium combinations.
TABLE 4 Effect of different hormone treatments on cotyledon callus and adventitious bud Induction
Note: lower case letters indicate significant differences at the P <0.05 level
5, the root is explant induced callus and adventitious bud
Comparing the effect of the hormone combination of 6-BA and TDZ with different concentrations on the induction of the cotyledon callus, carrying out variance analysis, and 20 hormone treatment results show that: the induction rate of the callus of a20(MS +30g/L Sucrose +7g/L Agar +5mg/L6-BA +1mg/L TDZ) culture medium reaches 50.71%, but no adventitious bud is induced by all the culture medium combinations.
TABLE 5 Effect of different hormone treatments on root callus and adventitious bud Induction
Note: lower case letters indicate significant differences at the P <0.05 level
6, the stem of the tissue culture seedling is used as an explant to induce callus and adventitious buds
Comparing the effect of the hormone combination of 6-BA and TDZ with different concentrations on the induction of the stem callus, carrying out variance analysis, and the results of 15 hormone treatments show that: the induction rates of the c7, c10, c12 and c14 calluses respectively reach 98.54%, 98.74%, 99.15% and 98.42% at the highest, and the differences are not significant. Wherein the induction rate of the adventitious bud of the c12(MSB5+30g/L sucrose +7g/L agar +5mg/L6-BA +8mg/L TDZ +2mg/L glycine +0.1mg/L IBA + 5% coconut water) culture medium is the highest, which reaches 33.33%, and the difference with the rest 14 treatments is obvious. The formed callus is hard inside, and small dark green projections are formed on the surface, and the projections can be converted into bud primordia to finally form adventitious buds.
TABLE 6 Effect of different hormone treatments on the induction of Stem callus and adventitious buds
Note: lower case letters indicate significant differences at the P <0.05 level
In conclusion, the inventor finds that the callus is easy to obtain from different tissues of the fraxinus mandshurica, but the callus state and adventitious bud induction difference obtained from different materials is obvious, and the adventitious bud induction rate of the stem section of the tissue culture seedling is the highest and reaches 33.33%. The callus induced by the leaves is hard, dark green, slow in growth speed, easy to age in internal tissues and not beneficial to the formation of bud primordium, so that the regenerated buds cannot be differentiated. The callus induced by the root has too loose texture, high growth speed and easy browning, and the callus after the light culture can not form small bud primordium bulges, so adventitious buds can not be formed. The callus formation and bud induction results of the two materials show that the hard slow-growing callus formed by the mature cotyledon tissue and the loose fast-growing callus formed by the tender root are not beneficial to the differentiation of adventitious buds.
In the process of culturing the hypocotyl and stem tissue callus, the callus with moderate hardness and slight looseness is formed, small dark green bulges can be formed on the tissue surface, gaps are formed among the lumps, and the survival of newly divided and differentiated cells is facilitated, so that bud primordium is formed, and finally adventitious buds are formed. In addition, compared with the callus formed by stem segments, the callus formed by hypocotyls is similar to cotyledon callus inside and is easy to age. The inside and the outside of the callus formed by the stem segments are more uniform, the growth speed of the callus is relatively slow, the texture is looser, the cell mass is fuller, the local substance accumulation of the callus is facilitated, the formation of bud primordium is facilitated, and finally the adventitious bud is formed through differentiation and growth.
Claims (1)
1. A method for inducing adventitious bud regeneration by fraxinus mandshurica tender green semi-compact callus is established, and the method is characterized in that:
(1) the explant is sourced: conventional surface sterilization, stripping mature embryos, culturing by WPM until cotyledons are unfolded, cutting hypocotyls, inoculating to a20 culture medium, and inducing callus and adventitious buds; cutting a non-bud stem section between two bud points of the fraxinus mandshurica aseptic seedling with the plant height of 3-5cm, and inoculating the non-bud stem section between the two bud points of the fraxinus mandshurica aseptic seedling to a c12 culture medium to induce callus and adventitious buds;
(2) callus and adventitious bud induction medium: the hypocotyl a20 culture medium contains MS +30g/L sucrose +7g/L agar +5mg/L6-BA +1mg/L TDZ; the stem section c12 medium contained MSB5+30g/L sucrose +7g/L agar +5mg/L6-BA +8mg/L TDZ +0.1mg/L IBA +2mg/L glycine + 5% coconut water, pH was 5.8;
(3) culture conditions and induction rate: the inducing material was cultured in dark for 14 days and then transferred to light culture with illumination intensity of 80 μmol m-2*s-116 h/8 h in dark, 25 +/-2 ℃ in temperature and 80-90% in humidity; callus induction rate at 20 d: the stem segment is 99.15 percent, and the hypocotyl is 88.69 percent; adventitious bud induction rate at 30 d: stem segment 33.33%, hypocotyl 10.52%;
(4) and (3) culture state: the hardness of the inside of the callus formed by the induction of the hypocotyl and the stem segment is moderate and is slightly loose, small dark green bulges can be formed on the surface, gaps are formed among the lumps, and the survival of newly divided and differentiated cells is facilitated, so that bud primordium is formed, and finally adventitious buds are formed;
(5) comparison of calli formed from hypocotyls and stem segments: the callus formed by hypocotyls is hard in internal texture, high in growth speed, dark green and easy to age; the callus formed by the stem segments is uniform inside and outside, has slightly loose texture compared with the hypocotyl, has slow growth speed, is yellow green, has fuller cell mass, is more favorable for the local substance accumulation of the callus, is favorable for the formation of bud primordium, finally differentiates and grows to form adventitious buds, most of the adventitious buds are clustered, and has good growth state after successive generations and high survival rate.
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Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USPP11777P2 (en) * | 1999-01-20 | 2001-02-13 | Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture | Hybrid ash tree named ‘Northern Gem’ |
| CN102888379A (en) * | 2012-11-06 | 2013-01-23 | 东北林业大学 | Method for establishing fraxinus mandshurica suspension culture system |
| CN104094840A (en) * | 2013-04-02 | 2014-10-15 | 东北林业大学 | Establishment method for Fraxinus rhynchophylla Hance. suspension culture system |
| CN109757382A (en) * | 2019-03-21 | 2019-05-17 | 鲁东大学 | The rapid propagation method of Chinese wax male elite plant strain |
| CN112106654A (en) * | 2019-06-12 | 2020-12-22 | 东北林业大学 | Method for obtaining fraxinus mandshurica regeneration plant through adventitious bud direct germination way |
| WO2021187995A2 (en) * | 2020-03-18 | 2021-09-23 | Uniwersytet Jagielloński | The method of obtaining saplings of the common ash (fraxinius excelsior l.) and the media suitable for use in this method |
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- 2019-08-07 CN CN201910725313.6A patent/CN111758566B/en not_active Expired - Fee Related
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| USPP11777P2 (en) * | 1999-01-20 | 2001-02-13 | Her Majesty The Queen In Right Of Canada, As Represented By The Minister Of Agriculture | Hybrid ash tree named ‘Northern Gem’ |
| CN102888379A (en) * | 2012-11-06 | 2013-01-23 | 东北林业大学 | Method for establishing fraxinus mandshurica suspension culture system |
| CN104094840A (en) * | 2013-04-02 | 2014-10-15 | 东北林业大学 | Establishment method for Fraxinus rhynchophylla Hance. suspension culture system |
| CN109757382A (en) * | 2019-03-21 | 2019-05-17 | 鲁东大学 | The rapid propagation method of Chinese wax male elite plant strain |
| CN112106654A (en) * | 2019-06-12 | 2020-12-22 | 东北林业大学 | Method for obtaining fraxinus mandshurica regeneration plant through adventitious bud direct germination way |
| WO2021187995A2 (en) * | 2020-03-18 | 2021-09-23 | Uniwersytet Jagielloński | The method of obtaining saplings of the common ash (fraxinius excelsior l.) and the media suitable for use in this method |
Non-Patent Citations (4)
| Title |
|---|
| Effects of Hormones and Epigenetic Regulation on the Callus and Adventitious Bud Induction of Fraxinus mandshurica Rupr.;Lei Yu等;《Forests》;20200524;第11卷;第1-18页 * |
| 水曲柳（Fraxinus mandshurica Rupr.）不定芽发生体系与发育机理的研究;黄剑;《中国优秀博士学位论文全文数据库（电子期刊）农业科技辑》;20101215(第12期);D049-29 * |
| 水曲柳子叶和下胚轴愈伤组织和体胚的诱导;张丽杰等;《分子植物育种》;20150728;第13卷(第7期);第1645-1652页 * |
| 水曲柳愈伤组织不定芽再生与FmWIND1基因克隆和表达研究;李新宇;《中国优秀硕士学位论文全文数据库（电子期刊）农业科技辑》;20200115(第1期);D049-344 * |
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