CN103688858B - A kind of oil palm plantlet in vitro root induction and short root strong seedling culture method - Google Patents
A kind of oil palm plantlet in vitro root induction and short root strong seedling culture method Download PDFInfo
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- CN103688858B CN103688858B CN201310692066.7A CN201310692066A CN103688858B CN 103688858 B CN103688858 B CN 103688858B CN 201310692066 A CN201310692066 A CN 201310692066A CN 103688858 B CN103688858 B CN 103688858B
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Abstract
The method of a kind of oil palm of the present invention plantlet in vitro root induction and short root strong seedling culture, aseptically, after oil palm unrooted plantlet in vitro conventional treatment, proceed in root induction medium and cultivate, Plantlet formation root system can be induced, and then be transferred in short root strong seedling culture base and carry out short root strong seedling culture, the oil palm plantlet in vitro whole plant of well developed root system stalwartness can be obtained.The present invention utilizes different medium and condition of culture to establish a set of workable, oil palm plantlet in vitro root induction that the strong plantlets and rootage cycle is short, cost-saving and short root strong seedling culture technology, healthy and strong flourishing root system can be obtained, can be applicable in the large-scale High-efficient Production of oil palm plantlet in vitro, for production oil of high quality palm fibre plantlet in vitro provides technical support, promote to there is the development that efficient oil palm plantlet in vitro is commercially produced realistic meaning.
Description
Technical field
The invention belongs to plant group technical field of tissue culture, relate to a kind of root induction and short root strong seedling culture method, specifically a kind of oil palm plantlet in vitro root induction and short root strong seedling culture method.
Background technology
Oil palm (Elaeis guineensis Jacq), Palmae (Palmae) oil palm belongs to perennial single growing point arbor, one of important oil crop in torrid areas, has the good reputation of " world oil king ".The grease yield of oil palm is extra-high, and it can be used as edible oil and fat, the raw material of industry, bioenergy etc., and purposes is relatively extensive, the huge market demand and gaining great popularity, and industry development is rapid, and therefore plantation oil palm is competitively introduced a fine variety in tropical and subtropical zone area all over the world.
Oil palm breeds and carries out mainly through seed, but is commonly heterozygote due to oil palm gene, and its offspring's genetic character is difficult to preserve parent's good characteristic, and population homogeneity is poor, moreover oil palm only has a growing point, is difficult to realize traditional clone and breeds.The nutrition organs such as blade are utilized to carry out tissue cultures, the excellent genetic character of parent both preserved by the Conditions In Relation To Oil Palm Cultivation material obtained, genetic character is homogeneous again, is to solve genetic character in oil palm seeling industry be separated and cannot realize the effective way that traditional clone breeds problem.
Root induction culture technique is one of core technology in plant tissue culture technique.Healthy and strong flourishing root system is the guarantee of training tissue culture seedling transplant survival, is one of measurement index of plantlet in vitro commercial quality, and therefore root induction and culture technique are one of keys obtaining high-quality seedling efficiently, are also one of guarantees of seedling high commodity rate.
Summary of the invention
The object of the invention is to provide a kind of method of oil palm plantlet in vitro root induction and short root strong seedling culture, utilize different medium and condition of culture, root induction and strong seedling culture are carried out to oil palm plantlet in vitro, healthy and strong flourishing root system can be obtained, for production oil of high quality palm fibre plantlet in vitro provides technical support, promote to there is the development that efficient oil palm plantlet in vitro is commercially produced realistic meaning.
The technical solution adopted in the present invention:
A method for oil palm plantlet in vitro root induction and short root strong seedling culture, its concrete steps are as follows:
1, aseptically, after the healthy and strong oil palm unrooted plantlet in vitro conventional treatment of more than diameter of stem 2mm, height of seedling more than 5cm, dark green leaf color, proceed in root induction medium and cultivate, condition of culture: temperature 25 ~ 30 DEG C, intensity of illumination 800 ~ 1200lx, illumination every day 12h; Cultivate 8 ~ 10 weeks, the root system (Fig. 1) of the long 0.5 ~ 3cm of Plantlet formation 1 ~ 13, root can be induced.Described root induction medium is medium based on WPM, and adds NAA0.1 ~ 5mg.L
-1, paclobutrazol 1mg.L
-1, sucrose 30 ~ 60g.L
-1, agar 7g.L
-1, active carbon 1g.L
-1, pH5.6 ~ 6.0.
2, the plant obtaining root long more than 1cm through Fiber differentiation is transferred in short root strong seedling culture base and carries out short root strong seedling culture, condition of culture: temperature 25 ~ 30 DEG C, intensity of illumination 2000 ~ 3000lx, illumination every day 16h; Short root strong seedling culture can obtain the oil palm plantlet in vitro whole plant (Fig. 2) of well developed root system stalwartness for 30 ~ 35 days.Described short root strong seedling culture base is medium based on WPM, and adds sucrose 75g.L
-1, pH5.6 ~ 6.0.Advantage of the present invention is:
(1) present invention employs two sections of root induction cultural methods and strict screening enter the root induction stage without offspring, make the culture materials root of hair time shorter, root of hair synchronism is high, and root growth is fast, seedling stalwartness.
(2) after the present invention adopts plant growth regulating substance startup to take root (long about the 1cm of root), namely proceed to the liquid nutrient medium without plant growth regulating substance, avoid plantlet in vitro and growing the negative effects such as the root growth that may occur is suppressed, leaf growth is not normal containing on the medium of higher plant growth regulating substance concentration for a long time.
(3) forward liquid nutrient medium to by seedling after the present invention adopts solid culture medium to start the root of induced synthesis long 1cm and realize short root strong sprout, fresh liquid medium Middle nutrition abundance exchanges fully, plantlet in vitro robust growth, well developed root system.
(4) the present invention adopts solid culture medium to start to forward liquid nutrient medium to by seedling after induction root forms long 1cm and realizes short root strong sprout, can reducing solid culture medium consumption, saving agar and growth regulatory substance consumption.Improve flourishing root system in seedling liquid medium within, there is not solid culture medium and glue the phenomenon twining root system, wash the seedling time after shortening seedling bottle outlet and reduce and wash the damage of seedling to root system.
The present invention establishes a set of workable, oil palm plantlet in vitro root induction that the strong plantlets and rootage cycle is short, cost-saving and short root strong seedling culture technology.The high-quality oil palm plantlet in vitro of well developed root system stalwartness can be obtained by this technology, can be applicable in the large-scale High-efficient Production of oil palm plantlet in vitro.
Accompanying drawing explanation
Fig. 1 is the root system that root induction cultivates the oil palm plantlet in vitro of cultivation after 9 weeks.
Fig. 2 is the oil palm plantlet in vitro of short root strong seedling culture well developed root system after 30 days.
Embodiment
Below in conjunction with embodiment, the specific embodiment of the present invention is described in further detail.Following examples for illustration of the present invention, but are not used for limiting the scope of the invention.The experimental technique of unreceipted actual conditions in the following example, usually conveniently condition, or according to the condition that manufacturer advises.
Embodiment one
1, root induction medium preparation: WPM+NAA0.1mg.L
-1+ paclobutrazol 1mg.L
-1+ sucrose 3g.L
-1+ agar 7g.L
-1+ active carbon 1g.L
-1, pH5.6.Often prop up test tube packing about 25 ~ 30ml medium, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.
2, choose diameter of stem 2 ~ 3mm, height of seedling 5 ~ 6cm the regeneration of healthy and strong oil palm leaf source somatic embryo without offspring 200 strain, cut-out blade and part stem segment base portion tissue, aseptically proceed to respectively in the root induction medium of step 1) preparation and cultivate, condition of culture: temperature 28 scholar 1 DEG C, intensity of illumination are about 1000lx, illumination every day 12h.Cultivate 8 weeks.Record data: strain number 128 strain of taking root, root length is 0.5 ~ 3cm.Root induction rate 64%.
3, short root strong seedling culture basigamy system: WPM+ sucrose 75g.L
-1, pH5.6.Often prop up the test tube packing liquid nutrient medium degree of depth about 1 ~ 2cm, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.Simultaneously described short root strong seedling culture base is except packing test tube, with the culture fluid of the many preparations of triangular flask packing, cultivates that liquid level is follow-up adds culture fluid in order to exposing with root elongation.
4, aseptically by step 2) in the seedling of long more than the 1cm of root of Fiber differentiation forward in short root strong seedling culture liquid and cultivate, add the fresh short root strong seedling culture liquid of step 3) preparation with root elongation in good time.Condition of culture: temperature is 28 scholar 1 DEG C, intensity of illumination 2500lx, illumination every day 16h.Short root strong seedling culture 30 days.Record data: the long 5cm of plantlet in vitro average root, generally grows one-level side root, form the oil palm plantlet in vitro whole plant of well developed root system stalwartness.
Embodiment two
1, root induction medium preparation: WPM+NAA1mg.L
-1+ paclobutrazol 1mg.L
-1+ sucrose 60g.L
-1+ agar 7g.L
-1+ active carbon 1g.L
-1, pH5.6.Often prop up test tube packing and be about 25ml medium, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.
2, choose diameter of stem 5 ~ 6mm, height of seedling 8 ~ 10cm the regeneration of healthy and strong oil palm leaf source somatic embryo without offspring 200 strain, after cut-out blade and part stem segment base portion organize, aseptically proceed to respectively in the root induction medium of step 1) preparation and cultivate, condition of culture: temperature 26 scholar 1 DEG C, intensity of illumination are about 1100lx, illumination every day 12h.Cultivate 10 weeks.Record data: strain number 184 strain of taking root, root length is 0.5 ~ 3cm.Root induction rate 92%.
3, short root strong seedling culture basigamy system: WPM+ sucrose 75g.L
-1, pH5.6.Often prop up the test tube packing liquid nutrient medium degree of depth about 1 ~ 2cm, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.Simultaneously described short root strong seedling culture base is except packing test tube, also uses the culture fluid of the many preparations of triangular flask packing, cultivates that liquid level is follow-up adds culture fluid in order to exposing with root elongation.
4, aseptically by step 2) in the seedling of long more than the 1cm of root of Fiber differentiation forward in short root strong seedling culture liquid and cultivate, add the fresh short root strong seedling culture base of step 3) preparation with root elongation in good time.Condition of culture: temperature is 27 scholar 1 DEG C, 2800lx, illumination every day 16h.Short root strong seedling culture 33 days.Record data: the long 8cm of plantlet in vitro average root, generally grows one-level side root, part grows secondary side root, forms the oil palm plantlet in vitro whole plant of well developed root system stalwartness.
Embodiment three
1, root induction medium preparation: WPM+NAA5mg.L
-1+ paclobutrazol 1mg.L
-1+ sucrose 60g.L
-1+ agar 7g.L
-1+ active carbon 1g.L
-1, pH6.0.Often prop up test tube packing and be about 25ml medium, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.
2, choose diameter of stem 5 ~ 6mm, height of seedling 8 ~ 10cm the regeneration of healthy and strong oil palm leaf source somatic embryo without offspring 200 strain, after cut-out blade and part stem segment base portion organize, aseptically proceed to respectively in the root induction medium of step 1) preparation and cultivate, condition of culture: temperature 27 scholar 2 DEG C, intensity of illumination are about 900lx, illumination every day 12h.Cultivate 9 weeks.Record data: strain number 173 strain of taking root, the long 0.5 ~ 3cm of root.Root induction rate 86.5%.
3, short root strong seedling culture basigamy system: WPM+ sucrose 75g.L
-1, pH5.6.Often prop up the test tube packing liquid nutrient medium degree of depth about 1 ~ 2cm, after adding a cover, in 1.2Kg/cm
3, sterilizing 20min under 121 DEG C of saturated vapour pressures.Described short root strong seedling culture base is except packing test tube simultaneously, also uses the culture fluid of the many preparations of triangular flask packing, and in order to exposing with root elongation, culture fluid in test tube is follow-up adds culture fluid.
4, aseptically by step 2) in the seedling of long more than the 1cm of root of Fiber differentiation forward in short root strong seedling culture liquid and cultivate, add the fresh short root strong seedling culture base of step 3) preparation with root elongation in good time.Condition of culture: temperature is 27 scholar 2 DEG C, 2200lx, illumination every day 16h.Short root strong seedling culture 32 days.Record data: the long 8cm of plantlet in vitro average root, generally grows one-level side root, part grows secondary side root, forms the oil palm plantlet in vitro whole plant of well developed root system stalwartness.
The above is only the preferred embodiment of the present invention; it should be pointed out that for those skilled in the art, under the prerequisite not departing from the technology of the present invention principle; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (1)
1. a method for oil palm plantlet in vitro root induction and short root strong seedling culture, it is characterized in that, concrete steps are as follows:
1), aseptically, after the healthy and strong oil palm unrooted plantlet in vitro conventional treatment of more than diameter of stem 2mm, height of seedling more than 5cm, dark green leaf color, proceed in root induction medium and cultivate, condition of culture: temperature 25 ~ 30 DEG C, intensity of illumination 800 ~ 1200lx, illumination every day 12h; Cultivate 8 ~ 10 weeks, induction Plantlet formation root system; Described root induction medium is medium based on WPM, and adds NAA0.1 ~ 5mg.L
-1, paclobutrazol 1mg.L
-1, sucrose 30 ~ 60g.L
-1, agar 7g.L
-1, active carbon 1g.L
-1, pH5.6 ~ 6.0;
2) plant, obtaining root long more than 1cm through Fiber differentiation is transferred in short root strong seedling culture base and carries out short root strong seedling culture, condition of culture: temperature 25 ~ 30 DEG C, intensity of illumination 2000 ~ 3000lx, illumination every day 16h; Short root strong seedling culture obtains the oil palm plantlet in vitro whole plant of well developed root system stalwartness for 30 ~ 35 days; Described short root strong seedling culture base is medium based on WPM, and adds sucrose 75g.L
-1, pH5.6 ~ 6.0.
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Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102715091A (en) * | 2012-07-06 | 2012-10-10 | 中国热带农业科学院椰子研究所 | Oil palm tissue culture breeding method |
| CN102860256A (en) * | 2012-07-16 | 2013-01-09 | 中国热带农业科学院橡胶研究所 | Oil palm tissue culture method |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN102715091A (en) * | 2012-07-06 | 2012-10-10 | 中国热带农业科学院椰子研究所 | Oil palm tissue culture breeding method |
| CN102860256A (en) * | 2012-07-16 | 2013-01-09 | 中国热带农业科学院橡胶研究所 | Oil palm tissue culture method |
Non-Patent Citations (2)
| Title |
|---|
| Optimizing of root induction in oil palm plantlets for acclimatization by some potent plant growth regulators (PGRs);Khairun Nizam et al;《Journal of Agricultural Technology》;20091231;第5卷(第2期);摘要，第373页第3段 * |
| 油棕组织培养;曾宪海等;《中国热带农业》;20121231(第48期);第2.3节，第6.2.1节 * |
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