CN100429305C - Cotton regenerated seedling rooting culture method and its special medium - Google Patents
Cotton regenerated seedling rooting culture method and its special medium Download PDFInfo
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- 229920000742 Cotton Polymers 0.000 title claims abstract description 49
- 238000012136 culture method Methods 0.000 title abstract description 4
- 229920001817 Agar Polymers 0.000 claims abstract description 12
- 239000008272 agar Substances 0.000 claims abstract description 12
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 12
- ZDXPYRJPNDTMRX-UHFFFAOYSA-N glutamine Natural products OC(=O)C(N)CCC(N)=O ZDXPYRJPNDTMRX-UHFFFAOYSA-N 0.000 claims abstract description 11
- 238000000034 method Methods 0.000 claims abstract description 10
- 239000000126 substance Substances 0.000 claims abstract description 8
- 201000009310 astigmatism Diseases 0.000 claims abstract description 4
- DHMQDGOQFOQNFH-UHFFFAOYSA-N Glycine Chemical compound NCC(O)=O DHMQDGOQFOQNFH-UHFFFAOYSA-N 0.000 claims description 8
- PVNIIMVLHYAWGP-UHFFFAOYSA-N Niacin Chemical compound OC(=O)C1=CC=CN=C1 PVNIIMVLHYAWGP-UHFFFAOYSA-N 0.000 claims description 8
- 239000004471 Glycine Substances 0.000 claims description 4
- 229960003512 nicotinic acid Drugs 0.000 claims description 4
- 235000001968 nicotinic acid Nutrition 0.000 claims description 4
- 239000011664 nicotinic acid Substances 0.000 claims description 4
- 229940064880 inositol 100 mg Drugs 0.000 claims description 3
- 229940029985 mineral supplement Drugs 0.000 claims 1
- 235000013619 trace mineral Nutrition 0.000 claims 1
- 239000011573 trace mineral Substances 0.000 claims 1
- 241000219146 Gossypium Species 0.000 abstract description 35
- 239000012882 rooting medium Substances 0.000 abstract description 14
- 239000002609 medium Substances 0.000 abstract description 7
- 229930006000 Sucrose Natural products 0.000 abstract description 6
- 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 abstract description 6
- 239000005720 sucrose Substances 0.000 abstract description 6
- 230000001651 autotrophic effect Effects 0.000 abstract description 4
- 239000001963 growth medium Substances 0.000 abstract description 4
- 238000009395 breeding Methods 0.000 abstract description 2
- 230000001488 breeding effect Effects 0.000 abstract description 2
- 230000001954 sterilising effect Effects 0.000 abstract 1
- 238000004659 sterilization and disinfection Methods 0.000 abstract 1
- 238000009423 ventilation Methods 0.000 abstract 1
- 239000000843 powder Substances 0.000 description 5
- 239000000654 additive Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 2
- 239000007864 aqueous solution Substances 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 239000007787 solid Substances 0.000 description 2
- SQUHHTBVTRBESD-UHFFFAOYSA-N Hexa-Ac-myo-Inositol Natural products CC(=O)OC1C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C(OC(C)=O)C1OC(C)=O SQUHHTBVTRBESD-UHFFFAOYSA-N 0.000 description 1
- 206010035148 Plague Diseases 0.000 description 1
- 241000607479 Yersinia pestis Species 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 239000012153 distilled water Substances 0.000 description 1
- CDAISMWEOUEBRE-GPIVLXJGSA-N inositol Chemical compound O[C@H]1[C@H](O)[C@@H](O)[C@H](O)[C@H](O)[C@@H]1O CDAISMWEOUEBRE-GPIVLXJGSA-N 0.000 description 1
- 229960000367 inositol Drugs 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 238000009630 liquid culture Methods 0.000 description 1
- 235000021073 macronutrients Nutrition 0.000 description 1
- 239000011785 micronutrient Substances 0.000 description 1
- 235000013369 micronutrients Nutrition 0.000 description 1
- 230000003020 moisturizing effect Effects 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 230000029553 photosynthesis Effects 0.000 description 1
- 238000010672 photosynthesis Methods 0.000 description 1
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- CDAISMWEOUEBRE-UHFFFAOYSA-N scyllo-inosotol Natural products OC1C(O)C(O)C(O)C(O)C1O CDAISMWEOUEBRE-UHFFFAOYSA-N 0.000 description 1
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Abstract
本发明公开了一种棉花再生苗生根培养方法及其专用培养基。棉花再生苗生根培养基水剂，每1000mL水中含有下述物质KNO3 1.9-3.5g，NH4NO3 0.65-1.7g，MgSO4 0.64-1.2g，KH2PO4 0.15-0.6g，CaCl2·H2O 0.3-1.2g，FeSO4·7H2O 0.2-0.4g，MnSO4·4H2O 0.017-0.085g，KI 0.004-0.02g，CuSO4 0.00003-0.00006g、CoCl20.00003-0.00006g、HBO3 0.007-0.0095g，NAA 2-10mg，KT 0.5-2mg，谷氨酰胺0.4-0.8g，琼脂6-8g，pH值为6.2-6.8。棉花再生苗生根培养方法，是将棉花再生苗生根培养基溶于水中，经高温灭菌后，冷却，将再生苗的基部插入培养基中，在20-25℃、散光下保湿培养12-18天，然后在光下通风培养。本发明在棉花的组织培养中将再生苗由人工添加蔗糖的异养培养转变为无蔗糖的自养培养方式，使棉花的生根移栽技术成为可能，将在棉花的工厂化繁育中发挥重要作用。The invention discloses a method for cultivating roots of regenerated cotton seedlings and a special culture medium thereof. Cotton regenerated seedling rooting medium water, each 1000mL water contains the following substances: KNO 3 1.9-3.5g, NH 4 NO 3 0.65-1.7g, MgSO 4 0.64-1.2g, KH 2 PO 4 0.15-0.6g, CaCl 2 H 2 O 0.3-1.2g, FeSO 4 7H 2 O 0.2-0.4g, MnSO 4 4H 2 O 0.017-0.085g, KI 0.004-0.02g, CuSO 4 0.00003-0.00006g, CoCl 2 0.00003-0.00006g , HBO 3 0.007-0.0095g, NAA 2-10mg, KT 0.5-2mg, glutamine 0.4-0.8g, agar 6-8g, pH value 6.2-6.8. The rooting culture method of cotton regenerated seedlings is to dissolve the rooting medium of cotton regenerated seedlings in water, after high-temperature sterilization, cool down, insert the base of regenerated seedlings into the medium, and keep moisture for 12-18 days at 20-25°C and astigmatism. day, and then cultivated under light and ventilation. In the tissue culture of cotton, the present invention transforms the regenerated seedlings from heterotrophic culture with artificial addition of sucrose to autotrophic culture without sucrose, making the rooting and transplanting technology of cotton possible, and will play an important role in the factory breeding of cotton .
Description
技术领域 technical field
本发明涉及农业组培技术领域中的一种棉花再生苗生根培养方法及其专用培养基。The invention relates to a rooting culture method for cotton regenerated seedlings and a special culture medium thereof in the technical field of agricultural tissue culture.
背景技术 Background technique
在棉花的组织培养技术中，再生苗的生根定植一直是困扰研究人员的一个世界性难题。目前，棉花再生苗的定植有嫁接和生根移栽两种方法。由于棉花再生苗生根培养困难，一般要经过液体培养才能诱导根的发生，并且成活率极低，故很少应用。嫁接是目前在棉花再生苗的定植中应用最多、最广的方法。但嫁接需要培育砧木、嫁接和嫁接后管理等繁琐步骤。In cotton tissue culture technology, the rooting and colonization of regenerated seedlings has always been a worldwide problem that plagues researchers. At present, there are two methods of planting cotton regenerated seedlings: grafting and rooting and transplanting. Because cotton regenerated seedlings are difficult to root and cultivate, it is generally necessary to induce the occurrence of roots through liquid culture, and the survival rate is extremely low, so it is rarely used. Grafting is currently the most widely used method in the field planting of cotton regenerated seedlings. However, grafting requires cumbersome steps such as cultivating rootstocks, grafting, and post-grafting management.
发明内容 Contents of the invention
本发明的目的是提供一种用于棉花组织培养的再生苗生根培养基。The object of the present invention is to provide a rooting medium for regenerated shoots for cotton tissue culture.
本发明所提供的棉花再生苗生根培养基水剂，每1000mL水中含有下述物质：KNO31.9-3.5g，NH4NO3 0.65-1.7g，MgSO4 0.64-1.2g，KH2PO4 0.15-0.6g，CaCl2·H2O0.3-1.2g，FeSO4·7H2O 0.02-0.04g，MnSO4·4H2O 0.017-0.085g，KI0.004-0.02g，CuSO4 0.00003-0.00006g、CoCl20.00003-0.00006g、HBO30.007-0.0095g，NAA 2-10mg，KT 0.5-2mg，谷氨酰胺0.4-0.8g，琼脂6-8g，pH值为6.2-6.8。The cotton regenerated seedling rooting medium aqueous solution provided by the present invention contains the following substances per 1000mL of water: KNO 3 1.9-3.5g, NH 4 NO 3 0.65-1.7g, MgSO 4 0.64-1.2g, KH 2 PO 4 0.15 -0.6g, CaCl 2 ·H 2 O 0.3-1.2g, FeSO 4 ·7H 2 O 0.02-0.04g, MnSO 4 ·4H 2 O 0.017-0.085g, KI0.004-0.02g, CuSO 4 0.00003-0.00006 g, CoCl 2 0.00003-0.00006g, HBO 3 0.007-0.0095g, NAA 2-10mg, KT 0.5-2mg, glutamine 0.4-0.8g, agar 6-8g, pH value 6.2-6.8.
优选的棉花再生苗生根培养基水剂配方为每1000mL水中含有下述物质：KNO32.85g，NH4NO3 1.47g，MgSO4 0.78g，KH2PO4 0.255g，CaCl2·H2O 0.667g，FeSO4·7H2O0.028g，MnSO4·4H2O 0.025g，KI 0.005g，CuSO4 0.00004g、CoCl20.00004g、HBO30.0093g，NAA 6mg，KT 2mg，谷氨酰胺0.6g，琼脂6.8g，pH值为6.5。The preferred formulation of cotton regenerated seedling rooting medium water contains the following substances per 1000mL of water: KNO 3 2.85g, NH 4 NO 3 1.47g, MgSO 4 0.78g, KH 2 PO 4 0.255g, CaCl 2 ·H 2 O 0.667g, FeSO 4 7H 2 O 0.028g, MnSO 4 4H 2 O 0.025g, KI 0.005g, CuSO 4 0.00004g, CoCl 2 0.00004g, HBO 3 0.0093g, NAA 6mg, KT 2mg, Glutamine 0.6 g, agar 6.8g, pH value is 6.5.
为得到较好的生根效果，所述棉花枝条扦插生根水剂中还添加有微量添加剂，包括下述微量添加剂中的一种或几种：甘氨酸2mg，VB10.4mg，VB60.5mg，烟酸0.5mg和肌醇100mg。In order to obtain a better rooting effect, trace additives are also added to the cotton branch cutting rooting water, including one or more of the following trace additives: 2 mg of glycine, 10.4 mg of VB, 0.5 mg of VB, 0.5 mg of nicotinic acid mg and inositol 100mg.
为方便包装，上述棉花再生苗生根培养基可为粉剂。所述棉花再生苗生根培养基粉剂中包含有重量份数比为1.9-3.5∶0.65-1.7∶0.64-1.2∶0.15-0.6∶0.3-1.2∶0.02-0.04∶0.017-0.085∶0.004-0.02∶0.00003-0.00006∶0.00003-0.00006∶0.007-0.0095∶0.002-0.01∶0.0005-0.002∶0.4-0.8∶6-8的KNO3、NH4NO3、MgSO4、KH2PO4、CaCl2·H2O、FeSO4·7H2O、MnSO4·4H2O、KI、CuSO4、CoCl2、HBO3、NAA、KT、谷氨酰胺和琼脂。For the convenience of packaging, the above-mentioned rooting medium for cotton regenerated seedlings can be a powder. The cotton regenerated seedling rooting medium powder contains a weight and number ratio of 1.9-3.5: 0.65-1.7: 0.64-1.2: 0.15-0.6: 0.3-1.2: 0.02-0.04: 0.017-0.085: 0.004-0.02: 0.00003 -0.00006: 0.00003-0.00006: 0.007-0.0095: 0.002-0.01: 0.0005-0.002: 0.4-0.8: 6-8 KNO 3 , NH 4 NO 3 , MgSO 4 , KH 2 PO 4 , CaCl 2 ·H 2 O, FeSO 4 .7H 2 O, MnSO 4 .4H 2 O, KI, CuSO 4 , CoCl 2 , HBO 3 , NAA, KT, Glutamine and Agar.
优选的棉花再生苗生根培养基粉剂配方包含有重量份数比为包含有重量份数比为2.85∶1.47∶0.78∶0.255∶0.667∶0.028∶0.025∶0.01∶0.00004∶0.00004∶0.0093∶0.006∶0.002∶0.6∶6.8的KNO3、NH4NO3、MgSO4、KH2PO4、CaCl2·H2O、FeSO4·7H2O、MnSO4·4H2O、KI、CuSO4、CoCl2、HBO3、NAA、KT、谷氨酰胺和琼脂。The preferred cotton regenerated seedling rooting medium powder formulation comprises a weight and number ratio of 2.85: 1.47: 0.78: 0.255: 0.667: 0.028: 0.025: 0.01: 0.00004: 0.00004: 0.0093: 0.006: 0.002: 0.6:6.8 KNO 3 , NH 4 NO 3 , MgSO 4 , KH 2 PO 4 , CaCl 2 ·H 2 O, FeSO 4 ·7H 2 O, MnSO 4 ·4H 2 O, KI, CuSO 4 , CoCl 2 , HBO 3. NAA, KT, glutamine and agar.
棉花再生苗生根培养基粉剂中也可添加有重量份数比为2∶0.4∶0.5∶0.5∶100的甘氨酸、VB1、VB6、烟酸或/和肌醇中的一种或几种微量添加剂。One or more trace additives of glycine, VB1, VB6, nicotinic acid or/and inositol can also be added in the rooting medium powder of cotton regenerated seedlings with a weight ratio of 2:0.4:0.5:0.5:100.
上述再生苗生根培养基水剂使用前只需加热将琼脂融化，冷却后即可。粉剂则还需先用水将其溶解，配成水剂。Before using the above-mentioned rooting medium water for regenerated seedlings, it only needs to be heated to melt the agar, and then it can be cooled. The powder also needs to be dissolved in water first to make an aqueous solution.
本发明的第二个目的提供一种用上述棉花再生苗生根培养基进行棉花再生苗生根培养的方法。The second object of the present invention provides a method for rooting cotton regenerated seedling culture with the above-mentioned cotton regenerated seedling rooting medium.
本发明所提供的棉花再生苗生根培养方法，是将再生苗的基部插入棉花再生苗生根培养基中，在20-25℃、散光下保湿培养12-18天(避免阳光直射)，然后在光下(可为阳光直射)通风培养。The rooting culture method of cotton regenerated seedlings provided by the present invention is to insert the base of regenerated seedlings into the rooting medium of cotton regenerated seedlings, and cultivate them in moisture for 12-18 days at 20-25° C. under astigmatism (avoiding direct sunlight), and then in light Under (can be direct sunlight) ventilated culture.
所述棉花再生苗生根培养基使用前要在125-128℃下，灭菌12分钟。The cotton regenerated seedling rooting medium should be sterilized at 125-128° C. for 12 minutes before use.
在实际应用中，为扩大生根面，可将枝条基部的削成楔形，以利于扩大生根面，达到壮根、壮苗的效果。In practical applications, in order to expand the rooting surface, the base of the branches can be cut into a wedge shape to facilitate the expansion of the rooting surface and achieve the effect of strong roots and seedlings.
本发明提供了一种适合棉花再生苗生根定植的自养型固体培养基，含有促进植物细胞生根生长的调节剂、植物生长发育所需的大量、微量营养元素及培养基固化剂。该培养基将再生苗从在人工添加蔗糖的培养基上的异养状态转变为靠自身光合作用生长、并可生根移栽的自然生长状态，故可称之为棉花再生苗自养型固体生根培养基。再生苗的生根效果良好，移栽成活率可达75.4％。本发明在棉花的组织培养中将再生苗由人工添加蔗糖的异养培养转变为无蔗糖的自养培养方式，使棉花的生根移栽技术成为可能，将在棉花的工厂化繁育中发挥重要作用。The invention provides an autotrophic solid medium suitable for the rooting and colonization of regenerated cotton seedlings, which contains a regulator for promoting plant cell rooting growth, macronutrients and micronutrients required for plant growth and development, and a medium solidifying agent. This medium changes the regenerated seedlings from the heterotrophic state on the artificially added sucrose medium to the natural growth state that grows by its own photosynthesis and can be rooted and transplanted, so it can be called autotrophic solid rooting of cotton regenerated seedlings Medium. The rooting effect of the regenerated seedlings is good, and the transplanting survival rate can reach 75.4%. In the tissue culture of cotton, the present invention transforms the regenerated seedlings from heterotrophic culture with artificial addition of sucrose to autotrophic culture without sucrose, making the rooting and transplanting technology of cotton possible, and will play an important role in the factory breeding of cotton .
具体实施方式 Detailed ways
下述实施例中所用方法如无特别说明均为常规方法。The methods used in the following examples are conventional methods unless otherwise specified.
实施例1、棉花再生苗生根培养基的配制Embodiment 1, the preparation of cotton regenerated seedling rooting medium
称取如下物质：KNO3 2.5g，NH4NO3 1.0g，MgSO4 0.78g，KH2PO4 0.3g，CaCl2·H2O0.8g，FeSO4·7H2O 0.028g，MnSO4·4H2O 0.05g，KI 0.01g，NAA 5mg，KT 1.0mg，琼脂6.8g，CuSO4 0.00004g，CoCl2 0.00004g，HBO3 0.0093g和谷氨酰胺0.5g。Weigh the following substances: KNO 3 2.5g, NH 4 NO 3 1.0g, MgSO 4 0.78g, KH 2 PO 4 0.3g, CaCl 2 H 2 O 0.8g, FeSO4 7H 2 O 0.028g, MnSO 4 4H 2 O 0.05 g, KI 0.01 g, NAA 5 mg, KT 1.0 mg, Agar 6.8 g, CuSO 4 0.00004 g, CoCl 2 0.00004 g, HBO 3 0.0093 g and Glutamine 0.5 g.
将上述物质溶于1升蒸馏水中，调pH值为6.5，分装到20个25mL的三角瓶中，每瓶50mL，用封口膜封口，125℃加热12分钟后，冷却备用。Dissolve the above-mentioned substances in 1 liter of distilled water, adjust the pH value to 6.5, divide into 20 25mL Erlenmeyer flasks, each bottle is 50mL, seal with parafilm, heat at 125°C for 12 minutes, and cool for later use.
实施例2、棉花再生苗的生根培养The rooting culture of embodiment 2, cotton regenerated seedling
在无菌的超净工作台中进行再生苗的转接，具体方法为：选用带有2-3片真叶的健壮无病的棉花再生苗，将基部削成约5mm长的楔形，扦插到实施例1制备的培养基中，对照为采用棉花再生苗常用的生长培养基(在MS基本培养基的基础上添加25g/L蔗糖、0.01mg/L NAA和0.005mg/L KT)在22℃、散光下保湿培养12-18天(避免阳光直射)，然后在光下(可为阳光直射)通风培养，约一个月后移栽，对照取生根量相当的棉苗移栽，结果用实施例1培养的再生苗成活率高达75.4％，对照全部死亡。The transfer of regenerated seedlings is carried out in a sterile ultra-clean workbench. The specific method is: select robust and disease-free cotton regenerated seedlings with 2-3 true leaves, cut the base into a wedge shape about 5 mm long, and cut the seedlings until the time of implementation. In the culture medium prepared in Example 1, the control is the growth medium (adding 25g/L sucrose, 0.01mg/L NAA and 0.005mg/L KT on the basis of MS basic medium) commonly used for cotton regenerated seedlings at 22°C, Moisturizing and cultivating under astigmatism for 12-18 days (avoiding direct sunlight), then ventilating and cultivating under light (can be direct sunlight), transplanting after about one month, and transplanting cotton seedlings with a comparable rooting amount as a control, the results are compared with those of Example 1. The survival rate of the cultivated regenerated shoots was as high as 75.4%, and all the control plants died.
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| CN111685123A (en) * | 2020-07-17 | 2020-09-22 | 深圳万禾农业科技有限公司 | Plant growth regulator composition and preparation method and application thereof |
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| Title |
|---|
| 玉米芽尖培养和丛生芽发生. 李学红，苏玲，张举仁.山东大学学报(自然科学版)，第32卷第4期. 1997 |
| 玉米芽尖培养和丛生芽发生. 李学红，苏玲，张举仁. 山东大学学报(自然科学版)，第32卷第4期. 1997 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102057869B (en) * | 2009-11-18 | 2012-09-26 | 中国农业科学院棉花研究所 | Cotton regeneration seedling rooting method and special culture medium thereof |
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| CN1908160A (en) | 2007-02-07 |
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