CN107810852A - 一种鼓槌石斛组织培养方法 - Google Patents
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- 244000283926 Dendrobium chrysotoxum Species 0.000 title claims abstract description 34
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- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims abstract description 16
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- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims abstract description 8
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- 229960003512 nicotinic acid Drugs 0.000 claims abstract description 8
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- ZGTMUACCHSMWAC-UHFFFAOYSA-L EDTA disodium salt (anhydrous) Chemical compound [Na+].[Na+].OC(=O)CN(CC([O-])=O)CCN(CC(O)=O)CC([O-])=O ZGTMUACCHSMWAC-UHFFFAOYSA-L 0.000 abstract description 5
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Classifications
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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
-
- 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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Abstract
本发明公开了一种鼓槌石斛组织培养方法,包括鼓槌石斛种子培养萌发成圆球茎,壮苗培养,诱导生根,炼苗四个步骤,所述鼓槌石斛种子培养萌发成圆球茎中使用的种子培养基包含以下成分:硝酸钾1800mg/L、硝酸铵1450mg/L、硫酸镁370mg/L、磷酸二氢钾170mg/L、氯化钙440mg/L、硫酸铜0.025mg/L、氯化钴0.025mg/L、硫酸锰16.9mg/L、硫酸锌8.6mg/L、硼酸6.2mg/L、碘化钾0.83mg/L、钼酸钠0.25mg/L、Na2‑EDTA 37.3mg/L、硫酸亚铁27.8mg/L、甘氨酸2.0mg/L、盐酸硫胺素0.1mg/L、盐酸吡哆醇0.5mg/L、烟酸0.5mg/L、肌醇100mg/L、NAA 1‑10mg/L、6‑BA 5‑10mg/L、马铃薯100000mg/L、碳粉250mg/L、蔗糖30000mg/L、水解乳蛋白700mg/L。该方法提高了鼓槌石斛的繁殖效率。
Description
技术领域
本发明涉及一种鼓槌石斛组织培养方法。
背景技术
鼓槌石斛(学名:Dendrobium chrysotoxum Lindl.)生于海拔520-1620米,阳光充足的常绿阔叶林中树干上或疏林下岩石上。分布于中国、印度、缅甸、泰国、老挝、越南。鼓槌石斛的花观赏价值极高,花姿优雅,玲珑可爱,花色鲜艳,气味芳香,被喻为“四大观赏洋花”之一,既可作切花,也可盆栽观赏,至今,广泛用于大型宴会、开幕式剪彩典礼或招待贵宾。此外,花朵剪下2-3天也不凋萎,生命力旺盛令人赞叹。
目前,虎牙石斛的人工培育包括分株繁殖、扦插繁殖、分芽繁殖,但是,上述繁殖方式繁殖效率低,不能满足市场需求,因此,需要利用植物组织培养技术对虎牙石斛进行快速繁殖。
发明内容
本发明的目的在于提供一种鼓槌石斛组织培养方法,该方法可以提高鼓槌石斛的繁殖效率。
为实现本发明的目的,本发明采用以下技术方案:一种鼓槌石斛组织培养方法,包括鼓槌石斛种子培养萌发成圆球茎,壮苗培养,诱导生根,炼苗四个步骤,其中,所述鼓槌石斛种子培养萌发成圆球茎中使用的种子培养基包含以下成分:
一、种子培养基配方
所述壮苗培养中使用的壮苗培养基包含以下成分:
二、壮苗培养基配方
试剂名称 | 规定量(mg/L) | 试剂名称 | 规定量(mg/L) |
硝酸钾 | 2000 | Na2-EDTA | 37.3 |
硝酸铵 | 1750 | 硫酸亚铁 | 27.8 |
硫酸镁 | 370 | 甘氨酸 | 2.0 |
磷酸二氢钾 | 170 | 盐酸硫胺素 | 0.2 |
氯化钙 | 550 | 盐酸吡哆醇 | 0.65 |
硫酸铜 | 0.025 | 烟酸 | 0.5 |
氯化钴 | 0.025 | 肌醇 | 50 |
硫酸锰 | 16.9 | NAA | 1-10 |
硫酸锌 | 8.6 | 6-BA | 5-10 |
硼酸 | 6.2 | 香蕉 | 200000 |
碘化钾 | 0.83 | 碳粉 | 500 |
钼酸钠 | 0.25 | 蔗糖 | 30000 |
所述诱导生根中使用的生根培养基包含以下成分:
三、生根培养基配方
试剂名称 | 规定量(mg/L) | 试剂名称 | 规定量(mg/L) |
硝酸钾 | 1650 | 硫酸亚铁 | 27.8 |
硝酸铵 | 1350 | 甘氨酸 | 2.0 |
硫酸镁 | 270 | 盐酸硫胺素 | 0.2 |
磷酸二氢钾 | 130 | 盐酸吡哆醇 | 0.65 |
氯化钙 | 450 | 烟酸 | 0.5 |
硫酸铜 | 0.025 | 肌醇 | 5 |
氯化钴 | 0.025 | NAA | 1-10 |
硫酸锰 | 16.9 | 6-BA | 0.1-10 |
硫酸锌 | 8.6 | 香蕉 | 100000 |
硼酸 | 6.2 | 碳粉 | 500 |
碘化钾 | 0.83 | 蔗糖 | 18000 |
钼酸钠 | 0.25 | 艾比蒂生根粉 | 0.1-1 |
Na2-EDTA | 37.3 |
所述种子培养萌发成圆球茎步骤中,成熟鼓槌石斛种子经消毒处理后播种于种子培养基中培养,一星期转绿,继续培养20-30天,经过种胚膨大、萌发形成绿色圆球茎;培养条件为25-26℃,光照为1600-1800lx,光照时间为10h/d。
所述的消毒处理过程为:自来水加洗洁精冲洗3分钟后用75%酒精消毒30秒,之后用35%双氧水消毒10分钟,然后用无菌水冲洗三次。
所述壮苗步骤中,圆球茎转入壮苗培养基中进行壮苗培养,经过80-90天壮苗培养,获得高度达到1.5-3cm的幼苗,培养条件:温度25-26℃,光照度1900-2100lx,光照时间为12h/d。
本发明具有以下优点:
1.本发明提供的鼓槌石斛的组织培养方法的鼓槌石斛种子萌发率高,在90%以上,获得的幼苗粗壮,主根系发达,种植成活率在90%以上,从而大大地提高了鼓槌石斛的繁殖效率。
2.种子培养基对鼓槌石斛种子的萌发有重要影响,本发明提供的种子培养基可以使鼓槌石斛种子萌发在90%以上。
3.本发明采用的壮苗培养基可以促进圆球茎生成粗壮的幼苗。
4.本发明采用的生根培养基可以促进幼苗生根,所得的主根系发达,有利于在炼苗及种植中成活。
具体实施方式
下面结合具体实施例对本发明做进一步的详细说明,但不构成对本发明的任何限制,任何在本发明权利要求范围内所做的有限修改,仍在本发明的权利要求保护范围内。
实施例1
鼓槌石斛组织培养分四个阶段,分别是种子萌发、壮苗培养、诱导生根和炼苗四个阶段,所以在本发明提供的鼓槌石斛组织培养方法包括鼓槌石斛种子萌发成为圆球茎,壮苗培养,生根,炼苗四个步骤,具体培养过程如下:
(1)种子萌发成为圆球茎:取成熟鼓槌石斛种子进行消毒处理,用自来水加洗洁精冲洗3分钟,然后75%酒精消毒30秒,再用35%双氧水消毒10分钟,最后用无菌水冲洗三次后播种于种子培养基中培养,一星期转绿,继续培养20-30天,经过种胚膨大、萌发形成绿色圆球茎,种子的萌发率在90%以上。培养条件为25-26℃,光照为1600-1800lx,光照时间为10h/d。
(2)壮苗培养:将步骤(1)中产生的圆球茎转入壮苗培养基中进行壮苗培养,经过80-90天壮苗培养,高度达到1.5-3cm的粗壮幼苗出现,培养条件:温度25-26℃,光照度1900-2100lx,光照时间为12h/d。
(3)诱导生根:将步骤(2)得到的幼苗转入生根培养基进行诱导生根,长成具有发达主根系的生根苗。
(4)炼苗:将步骤(3)得到的生根苗进行炼苗驯化,使其适应外部生长环境。完成炼苗后,所得驯化苗移栽大棚或室外,同时种植管理,驯化苗成活率在90%以上。
一、种子培养基配方
试剂名称 | 规定量(mg/L) | 试剂名称 | 规定量(mg/L) |
硝酸钾 | 1800 | 硫酸亚铁 | 27.8 |
硝酸铵 | 1450 | 甘氨酸 | 2.0 |
硫酸镁 | 370 | 盐酸硫胺素 | 0.1 |
磷酸二氢钾 | 170 | 盐酸吡哆醇 | 0.5 |
氯化钙 | 440 | 烟酸 | 0.5 |
硫酸铜 | 0.025 | 肌醇 | 100 |
氯化钴 | 0.025 | NAA | 1-10 |
硫酸锰 | 16.9 | 6-BA | 5-10 |
硫酸锌 | 8.6 | 马铃薯 | 100000 |
硼酸 | 6.2 | 碳粉 | 250 |
碘化钾 | 0.83 | 蔗糖 | 30000 |
钼酸钠 | 0.25 | 水解乳蛋白 | 700 |
Na2-EDTA | 37.3 |
二、壮苗培养基配方
试剂名称 | 规定量(mg/L) | 试剂名称 | 规定量(mg/L) |
硝酸钾 | 2000 | Na2-EDTA | 37.3 |
硝酸铵 | 1750 | 硫酸亚铁 | 27.8 |
硫酸镁 | 370 | 甘氨酸 | 2.0 |
磷酸二氢钾 | 170 | 盐酸硫胺素 | 0.2 |
氯化钙 | 550 | 盐酸吡哆醇 | 0.65 |
硫酸铜 | 0.025 | 烟酸 | 0.5 |
氯化钴 | 0.025 | 肌醇 | 50 |
硫酸锰 | 16.9 | NAA | 1-10 |
硫酸锌 | 8.6 | 6-BA | 5-10 |
硼酸 | 6.2 | 香蕉 | 200000 |
碘化钾 | 0.83 | 碳粉 | 500 |
钼酸钠 | 0.25 | 蔗糖 | 30000 |
三、生根培养基配方
种子培养基中的NAA的规定量在1-10mg/L范围内调整,6-BA的规定量在5-10mg/L范围内调整,都能够使鼓槌石斛种子萌发率达到90%以上。如,NAA的规定量为1mg/L、1.5mg/L、2mg/L、2.5mg/L、3mg/L、3.5mg/L、4mg/L、4.5mg/L、5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。6-BA的规定量为5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。
壮苗培养基中的NAA的规定量在1-10mg/L范围内调整,6-BA的规定量在5-10mg/L范围内调整,都能够使鼓槌石斛的圆球茎长出粗壮的幼苗。如,NAA的规定量为1mg/L、1.5mg/L、2mg/L、2.5mg/L、3mg/L、3.5mg/L、4mg/L、4.5mg/L、5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。6-BA的规定量为5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。
生根培养基中的NAA的规定量在1-10mg/L范围内调整,6-BA的规定量在5-10mg/L范围内调整,艾比蒂生根粉的规定量在0.1-1mg/L范围内调整,幼苗长出发达的主根系。如,NAA的规定量为1mg/L、1.5mg/L、2mg/L、2.5mg/L、3mg/L、3.5mg/L、4mg/L、4.5mg/L、5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。6-BA的规定量为5mg/L、5.5mg/L、6mg/L、6.5mg/L、7mg/L、7.5mg/L、8mg/L、8.5mg/L、9mg/L、9.5mg/L和10mg/L均可。艾比蒂生根粉的规定量为0.1mg/L、0.2mg/L、0.3mg/L、0.4mg/L、0.5mg/L、0.6mg/L、0.7mg/L、0.8mg/L、0.9mg/L、1mg/L均可。
Claims (6)
1.一种鼓槌石斛组织培养方法,包括鼓槌石斛种子培养萌发成圆球茎,壮苗培养,诱导生根,炼苗四个步骤,其特征是,所述鼓槌石斛种子培养萌发成圆球茎中使用的种子培养基包含以下成分:
硝酸钾1800mg/L、硝酸铵1450mg/L、硫酸镁370mg/L、磷酸二氢钾170mg/L、氯化钙440mg/L、硫酸铜0.025mg/L、氯化钴0.025mg/L、硫酸锰16.9mg/L、硫酸锌8.6mg/L、硼酸6.2mg/L、碘化钾0.83mg/L、钼酸钠0.25mg/L、Na2-EDTA 37.3mg/L、硫酸亚铁27.8mg/L、甘氨酸2.0mg/L、盐酸硫胺素0.1mg/L、盐酸吡哆醇0.5mg/L、烟酸0.5mg/L、肌醇100mg/L、NAA 1-10mg/L、6-BA 5-10mg/L、马铃薯100000mg/L、碳粉250mg/L、蔗糖30000mg/L、水解乳蛋白700mg/L。
2.根据权利要求1所述的鼓槌石斛组织培养方法,其特征是,所述壮苗培养中使用的壮苗培养基包含以下成分:硝酸钾2000mg/L、硝酸铵1750mg/L、硫酸镁370mg/L、磷酸二氢钾170mg/L、氯化钙550mg/L、硫酸铜0.025mg/L、氯化钴0.025mg/L、硫酸锰16.9mg/L、硫酸锌8.6mg/L、硼酸6.2mg/L、碘化钾0.83mg/L、钼酸钠0.25mg/L、Na2-EDTA 37.3mg/L、硫酸亚铁27.8mg/L、甘氨酸2.0mg/L、盐酸硫胺素0.2mg/L、盐酸吡哆醇0.65mg/L、烟酸0.5mg/L、肌醇50mg/L、NAA 1-10mg/L、6-BA 5-10mg/L、香蕉200000mg/L、碳粉500mg/L、蔗糖30000mg/L。
3.根据权利要求1或2所述的鼓槌石斛组织培养方法,其特征是,所述诱导生根中使用的生根培养基包含以下成分:硝酸钾1650mg/L、硝酸铵1350mg/L、硫酸镁270mg/L、磷酸二氢钾130mg/L、氯化钙450mg/L、硫酸铜0.025mg/L、氯化钴0.025mg/L、硫酸锰16.9mg/L、硫酸锌8.6mg/L、硼酸6.2mg/L、碘化钾0.83mg/L、钼酸钠0.25mg/L、Na2-EDTA 37.3mg/L、硫酸亚铁27.8mg/L、甘氨酸2.0mg/L、盐酸硫胺素0.2mg/L、盐酸吡哆醇0.65mg/L、烟酸0.5mg/L、肌醇5mg/L、NAA 1-10mg/L、6-BA 0.1-10mg/L、香蕉100000mg/L、碳粉500mg/L、蔗糖30000mg/L、艾比蒂生根粉0.1-1mg/L。
4.根据权利要求3所述的鼓槌石斛组织培养方法,其特征是,所述种子培养萌发成圆球茎步骤中,成熟鼓槌石斛种子经消毒处理后播种于种子培养基中培养,一星期转绿,继续培养20-30天,经过种胚膨大、萌发形成绿色圆球茎;培养条件为25-26℃,光照为1600-1800lx,光照时间为10h/d。
5.根据权利要求4所述的鼓槌石斛组织培养方法,其特征是,所述的消毒处理过程为:自来水加洗洁精冲洗3分钟后用75%酒精消毒30秒,之后用35%双氧水消毒10分钟,然后用无菌水冲洗三次。
6.根据权利要求4所述的鼓槌石斛组织培养方法,其特征是,所述壮苗步骤中,圆球茎转入壮苗培养基中进行壮苗培养,经过80-90天壮苗培养,获得高度达到1.5-3cm的幼苗,培养条件:温度25-26℃,光照度1900-2100lx,光照时间为12h/d。
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Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112106644A (zh) * | 2020-08-12 | 2020-12-22 | 德江县山顶中药材种植有限公司 | 一种铁皮石斛杂交品种选育及仿野生种植方法 |
Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1608424A (zh) * | 2004-11-24 | 2005-04-27 | 上海增靓生物科技有限公司 | 铁皮石斛的人工大面积栽培方法 |
CN101213940A (zh) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | 石斛属的快速繁殖方法 |
CN101569287A (zh) * | 2009-06-02 | 2009-11-04 | 中国科学院华南植物园 | 接瓣兰种苗的试管繁殖方法 |
CN101933456A (zh) * | 2009-06-29 | 2011-01-05 | 西双版纳增靓生物科技有限公司 | 铁皮石斛蒴果快繁育苗方法 |
CN104322371A (zh) * | 2014-10-29 | 2015-02-04 | 安徽牧龙山生态旅游开发股份有限公司 | 一种铁皮石斛组织培养培养基及其铁皮石斛组织培养方法 |
CN105052753A (zh) * | 2015-09-29 | 2015-11-18 | 江苏农林职业技术学院 | 一种铁皮石斛的快速繁殖方法 |
US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
-
2017
- 2017-11-02 CN CN201711112798.9A patent/CN107810852A/zh active Pending
Patent Citations (7)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN1608424A (zh) * | 2004-11-24 | 2005-04-27 | 上海增靓生物科技有限公司 | 铁皮石斛的人工大面积栽培方法 |
CN101213940A (zh) * | 2008-01-18 | 2008-07-09 | 中国科学院昆明植物研究所 | 石斛属的快速繁殖方法 |
CN101569287A (zh) * | 2009-06-02 | 2009-11-04 | 中国科学院华南植物园 | 接瓣兰种苗的试管繁殖方法 |
CN101933456A (zh) * | 2009-06-29 | 2011-01-05 | 西双版纳增靓生物科技有限公司 | 铁皮石斛蒴果快繁育苗方法 |
US20160143236A1 (en) * | 2013-07-08 | 2016-05-26 | South China Botanical Garden, Chinese Academy Of Sciences | Dendrobium in vitro crossbreeding method |
CN104322371A (zh) * | 2014-10-29 | 2015-02-04 | 安徽牧龙山生态旅游开发股份有限公司 | 一种铁皮石斛组织培养培养基及其铁皮石斛组织培养方法 |
CN105052753A (zh) * | 2015-09-29 | 2015-11-18 | 江苏农林职业技术学院 | 一种铁皮石斛的快速繁殖方法 |
Non-Patent Citations (1)
Title |
---|
蒙阳等: "始兴石斛的无菌播种及快速繁殖", 《南昌大学学报(理科版)》 * |
Cited By (1)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN112106644A (zh) * | 2020-08-12 | 2020-12-22 | 德江县山顶中药材种植有限公司 | 一种铁皮石斛杂交品种选育及仿野生种植方法 |
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