WO2015096463A1 - Milieu de croissance par induction de phalaenopsis et procédé de reproduction asexuée de phalaenopsis - Google Patents

Milieu de croissance par induction de phalaenopsis et procédé de reproduction asexuée de phalaenopsis Download PDF

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Publication number
WO2015096463A1
WO2015096463A1 PCT/CN2014/082190 CN2014082190W WO2015096463A1 WO 2015096463 A1 WO2015096463 A1 WO 2015096463A1 CN 2014082190 W CN2014082190 W CN 2014082190W WO 2015096463 A1 WO2015096463 A1 WO 2015096463A1
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WIPO (PCT)
Prior art keywords
phalaenopsis
concentration
induction medium
induction
gibberellin
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PCT/CN2014/082190
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English (en)
Chinese (zh)
Inventor
陈永得
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佛山市顺德区今日景艺生物科技有限公司
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Publication of WO2015096463A1 publication Critical patent/WO2015096463A1/fr

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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
    • A01H4/002—Culture media for tissue culture

Definitions

  • the invention relates to the field of plant tissue culture, in particular to a Phalaenopsis induction medium and a vegetative propagation method of Phalaenopsis. Background technique
  • Phalaenopsis Bl. spp. is a tropical gas blue orchid with large flowers, long flowering period, beautiful flowers, rich color, beautiful flower shape, such as butterfly dancing, hence the name. It is known as the "Queen of Orchids" in the tropics and is one of the most popular orchids in recent years. Phalaenopsis is a single-flowered aerial orchid. Plants rarely develop lateral buds, and seeds are extremely difficult to germinate. They are routinely propagated and the proliferation rate is very slow. Therefore, many species are rapidly propagated by tissue culture.
  • the object of the present invention is to provide a Phalaenopsis inducing medium and a method for vegetative propagation of Phalaenopsis, which can be used for inducing rapid propagation of Phalaenopsis in the internode of the butterfly orchid stem. Designed to provide a new method of asexual reproduction of Phalaenopsis.
  • a Phalaenopsis induction medium wherein the Phalaenopsis induction medium is prepared by mixing 6-nonylaminoadenine BA, gibberellin GA 3 and organic additives with a standard N6 medium-based mother liquor; -
  • the concentration of aminoguanidine BA is 0 ⁇ 10g/L
  • the concentration of gibberellin GA 3 is 0 ⁇ 1.5 mg/L
  • the concentration of organic additive is 0 ⁇ 300 g/L
  • 6- ⁇ aminoadenine The content of BA and gibberellin GA 3 is non-zero.
  • the phalaenopsis induction medium wherein the concentration of 6-nonamino adenine BA is 3 ⁇ 5 mg/L, the concentration of gibberellin GA 3 is l ⁇ 1.2 mg/L, and the concentration of organic additive is 150 ⁇ 180 g/L.
  • the Phalaenopsis induction medium has a concentration of 6 mg/ammonium adenine BA of 5 mg/L, a concentration of gibberellin GA 3 of 1.2 mg/L, and an organic additive concentration of 150 g/L.
  • the phalaenopsis induction medium wherein the phalaenopsis induction medium is added with agar, a carbon source and activated carbon; wherein the concentration of the agar is 7-9 g/L, and the concentration of the carbon source is 20-30 g/ L, the concentration of activated carbon is 1 ⁇ 3 g/L.
  • the phalaenopsis induction medium wherein the organic additive is coconut juice or tomato juice.
  • the phalaenopsis induction medium, wherein the organic additive is tomato juice.
  • the Phalaenopsis induction medium wherein the carbon source is white sugar or sucrose.
  • the phalaenopsis induction medium wherein the carbon source is white granulated sugar.
  • a method for vegetative propagation of Phalaenopsis wherein the pheasant vegetative propagation method is an explant with a peduncle internode, and the phalaenopsis induction medium as described above is used as an induction medium.
  • the pheasant vegetative propagation method wherein the phalaenopsis vegetative propagation method comprises the following steps:
  • the present invention provides a Phalaenopsis inducing medium and a method for vegetative propagation of Phalaenopsis.
  • the Phalaenopsis inducing medium can be used for inducing rapid propagation of Phalaenopsis in the internode of the butterfly orchid stem, which has low cost and protocorm-like bulbs.
  • the induction rate is high, the induction time is short, and the growth rate is fast.
  • the method for vegetative propagation of Phalaenopsis provided by the present invention is that the young peduncle is an explant and the phalaenopsis induction medium is used to rapidly propagate the phalaenopsis.
  • the peduncle internode is the waste in the traditional tissue culture production. This method can not only make full use of the peduncle material, but also turn waste into treasure, saving cost, and Flowers that are in the lower part of the peduncle can still be used for cross-breeding without damaging the female parent. detailed description
  • the present invention provides a Phalaenopsis Inducing Medium and a Phalaenopsis vegetative propagation method, and the present invention will be further described in detail below in order to clarify the purpose, technical solutions and effects of the present invention. It is understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
  • the present invention provides a Phalaenopsis induction medium for inducing rapid propagation of Phalaenopsis in the internodes of the Phalaenopsis orchid.
  • the Phalaenopsis induction medium is optimized by adding a plant growth regulator and an organic additive based on a standard N6 medium.
  • the plant growth regulator is 6-nonylaminoadenine BA and gibberellin GA 3 .
  • 6-nonylaminoadenine BA to the Phalaenopsis induction medium significantly promoted the induction and proliferation of protocorms in a certain range (0-5 mg/L, and the content is non-zero).
  • the dominant factor of the protocorm-like stems induced by the peduncle can increase the induction rate and the late proliferation coefficient.
  • Gibberellin GA 3 is a kind of phytohormone, which is mainly used to promote the growth of stems and leaves of plants, break the dormancy, promote heading, increase the number of male flowers in melons, induce parthenocarpy, and is rarely used in the induction of pedicels.
  • the gibberellin GA 3 was creatively added to the Phalaenopsis induction medium, which significantly shortened the induction time and induced protocorm-like bulbs 10 to 15 days earlier than the no-addition treatment.
  • the organic additive may be tomato juice or coconut juice, and in the solution of the present invention, preferably tomato juice.
  • tomato juice is a natural organic additive that prevents the use of high concentrations of growth regulators and causes protocorm to mutate in proliferation.
  • tomatoes are widely available all year round, everywhere, and cheap, reducing the cost of tissue culture.
  • the Phalaenopsis induction medium is prepared by mixing 6-nonylaminoadenine BA, gibberellin GA 3 and an organic additive based on a standard N6 medium; wherein, 6-fluorenylamino The concentration of adenine BA is 0 ⁇ 10g/L, the concentration of gibberellin GA 3 is 0 ⁇ 1.5 mg/L, the concentration of organic additive is 0 ⁇ 300 g/L; and 6- ⁇ aminoadenine BA and red The content of the GA 3 is non-zero.
  • the concentration of 6-mercaptoaminoadenine BA is 3 ⁇ 5 mg/L
  • the concentration of gibberellin GA 3 is l ⁇ 1.2 mg/L
  • the concentration of tomato juice is 150 ⁇ 180 g/L.
  • the concentration of 6-mercaptoaminoadenine BA is 5 mg/L
  • the concentration of gibberellin GA 3 is 1.2 mg/L
  • the concentration of tomato juice is 150 g/L.
  • the Phalaenopsis induction medium in this composition range has the highest induction rate, the shortest induction time, and the growth of protocorm-like bulbs is the best.
  • the phalaenopsis induction medium may further contain agar, a carbon source and activated carbon; wherein the concentration of the agar is 7 to 9 g/L, the concentration of the carbon source is 20 to 30 g/L, and the concentration of the activated carbon It is 1 ⁇ 3 g/L.
  • the carbon source may be white sugar or sucrose.
  • white sugar is preferably used as a carbon source, because in the Phalaenopsis induction medium, white sugar is used as a carbon source in the induction rate and the induction time. Reduce costs without being affected.
  • Agar can provide nutrients, moisture, and ventilating effect. The agar replaces the carrageenan in the traditional medium, and the preparation of the medium is reduced under the premise that the protocorm-inducing rate and the induction time are not affected. cost.
  • the moth orchid induction medium provided by the present invention can be used in addition to various nutrient mother liquors in distilled water, and the rest can use tap water instead of distilled water, thereby indirectly simplifying the tissue culture procedure and cost.
  • the invention also provides a method for vegetative propagation of Phalaenopsis, comprising the following steps:
  • the method for asexual reproduction of Phalaenopsis adopts the young peduncle as an explant, which not only fully utilizes the peduncle material, but also does not damage the female parent, and the flower at the lower part of the peduncle can also be used for hybridization;
  • the above-mentioned phalaenopsis induction medium is used as an induction medium, which has the advantages of low cost, high protocorm-inducing rate, short induction time, and fast growth rate.
  • the base mother liquor of the Phalaenopsis induction medium was prepared according to the composition and concentration of the standard N6 medium, and was dispensed into nine triangular flasks and labeled separately.
  • the No. 1 to No. 8 flasks were experimentally added to the Phalaenopsis induction medium supplemented with plant growth regulators and organic additives, and the No. 9 flask was a medium control group in which only agar, white granulated sugar and activated carbon were added.
  • Example 2 Inoculation and culture of peduncle internode Inoculation in a sterile ultra-clean workbench, using a sterilized scalpel to cut the incision in contact with the disinfectant at both ends of the peduncle, taking the young part between the two pedicel sections, and cutting into 2 ⁇ 3mm sections, quickly inoculation
  • the stopper was sealed with a sterilized stopper.
  • the culture flask After inoculation, the culture flask is placed in the artificial culture chamber.
  • the environmental parameters of the culture flask are as follows: Culture conditions: temperature 25 ⁇ 1 °C:, light intensity 1500 ⁇ 18001x, light and dark alternate 12h/12h, culture for 50 days, The induction rate, induction time and growth of the protocorm-like bulbs in the culture flask were recorded (see Table 1 for experimental data).
  • the induction rate effect and growth condition of the induction medium of the present invention are better, especially when a certain amount (3 ⁇ 5 mg/L) of 6- ⁇ is added.
  • the amino adenine BA Induction medium No. 1, No. 2, No. 5, No. 7, No. 8
  • the induction rate not only did not decrease, but also increased; in the control group, no 6-mercaptoaminoadenine BA was added.
  • the peduncle did not germinate during the internodes, and no protocorm-like bulbs were induced.
  • 6-aminoamino adenine BA can effectively increase the induction rate of protocorm-like bulbs, which is the key to the success of pedicel internodes.
  • Adding tomato juice has a certain promoting effect on the growth and development of protocorm-like bulbs, but the effect on the induction rate The noise is not obvious.
  • gibberellin GA 3 can significantly shorten the induction time without affecting the induction rate, 10 to 15 days earlier than the non-addition.

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  • Life Sciences & Earth Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Biotechnology (AREA)
  • Developmental Biology & Embryology (AREA)
  • Cell Biology (AREA)
  • Botany (AREA)
  • Environmental Sciences (AREA)
  • Breeding Of Plants And Reproduction By Means Of Culturing (AREA)
  • Cultivation Of Plants (AREA)

Abstract

L'invention porte sur un milieu de croissance par induction de Phalaenopsis et sur un procédé de reproduction asexuée de Phalaenopsis. Le milieu de croissance par induction de Phalaenopsis est préparé par mélange de milieu de croissance N6 standard servant de liqueur mère de base avec de la 6-benzylaminopurine (BA), de la gibbérelline A3 (GA3) et un additif organique ajouté. Le milieu de croissance par induction de Phalaenopsis peut être utilisé pour l'induction d'une partie d'entrenœud de pédoncule pour reproduire rapidement Phalaenopsis et a une vitesse élevée d'induction de corps de type protocorme et un court temps d'induction. La présente invention porte également sur un procédé de reproduction asexuée de Phalaenopsis, qui utilise un entrenœud de pédoncule nouvellement bourgeonné comme explant et emploie le milieu de croissance par induction de Phalaenopsis. Le procédé non seulement permet l'utilisation totale d'un matériau de pédoncule, mais de plus ne provoque pas d'endommagement sur une plante mère, une fleur située au niveau des parties inférieures du pédoncule pouvant être encore utilisée pour l'hybridation.
PCT/CN2014/082190 2013-12-27 2014-07-15 Milieu de croissance par induction de phalaenopsis et procédé de reproduction asexuée de phalaenopsis WO2015096463A1 (fr)

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CN201310746547.1A CN103766218B (zh) 2013-12-27 2013-12-27 一种蝴蝶兰诱导培养基及蝴蝶兰无性繁殖方法
CN201310746547.1 2013-12-27

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Cited By (4)

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CN109618919A (zh) * 2019-02-14 2019-04-16 芜湖东源新农村开发股份有限公司 一种提高展状樱桃蝴蝶兰开花率的栽培方法
CN110999792A (zh) * 2019-12-30 2020-04-14 临沧市云瑞堂生物科技有限公司 一种金线莲多芽培养方法
CN113229147A (zh) * 2021-06-07 2021-08-10 芜湖东源新农村开发股份有限公司 利用极限光照培育蝴蝶兰苗分化芽的方法
CN115104535A (zh) * 2022-07-20 2022-09-27 浙江农林大学 一种以叶片为外植体的蝴蝶兰再生为完整植株的方法

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CN104604687B (zh) * 2015-01-29 2017-07-18 赤峰市农牧科学研究院 利用切芽后的花梗茎段诱导丛生芽快速繁殖蝴蝶兰的方法
CN106489461A (zh) * 2015-09-07 2017-03-15 湖南瑞利农业发展有限公司 一种蝴蝶兰花梗苗的培育方法
CN106069778A (zh) * 2016-07-31 2016-11-09 张玉薇 春兰的组织培养方法
CN106234218A (zh) * 2016-07-31 2016-12-21 张玉薇 春兰组织培养用培养基

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN109618919A (zh) * 2019-02-14 2019-04-16 芜湖东源新农村开发股份有限公司 一种提高展状樱桃蝴蝶兰开花率的栽培方法
CN110999792A (zh) * 2019-12-30 2020-04-14 临沧市云瑞堂生物科技有限公司 一种金线莲多芽培养方法
CN113229147A (zh) * 2021-06-07 2021-08-10 芜湖东源新农村开发股份有限公司 利用极限光照培育蝴蝶兰苗分化芽的方法
CN115104535A (zh) * 2022-07-20 2022-09-27 浙江农林大学 一种以叶片为外植体的蝴蝶兰再生为完整植株的方法
CN115104535B (zh) * 2022-07-20 2023-09-19 浙江农林大学 一种以叶片为外植体的蝴蝶兰再生为完整植株的方法

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