WO2021103168A1 - 一种利用水杨酸促进杉木体胚发生的方法 - Google Patents

一种利用水杨酸促进杉木体胚发生的方法 Download PDF

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WO2021103168A1
WO2021103168A1 PCT/CN2019/125020 CN2019125020W WO2021103168A1 WO 2021103168 A1 WO2021103168 A1 WO 2021103168A1 CN 2019125020 W CN2019125020 W CN 2019125020W WO 2021103168 A1 WO2021103168 A1 WO 2021103168A1
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salicylic acid
fir
chinese fir
somatic
somatic embryogenesis
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PCT/CN2019/125020
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French (fr)
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陈金慧
施季森
陆叶
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南京林业大学
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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
    • A01H4/005—Methods for micropropagation; Vegetative plant propagation using cell or tissue culture techniques
    • A01H4/006—Encapsulated embryos for plant reproduction, e.g. artificial seeds

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  • the invention belongs to the technical field of plant tissue culture, and more specifically, relates to a method for promoting somatic embryogenesis of Chinese fir by using salicylic acid.
  • Chinese fir (Cunninghina lanceolata) belongs to the Chinese fir genus. It is one of the important timber species in my country. It grows fast, has a straight and round stem, is light and soft, easy to process, beautiful in texture, has a special aroma, does not warp or crack, Moth-resistant and corrosion-resistant, it is an excellent raw material for construction, furniture, bridges, shipbuilding, papermaking and wood fiber industries.
  • Chinese fir seedlings are mainly propagated through three methods: seeds, cuttings and traditional tissue culture [Wen Yafeng. Research on the induction and subgeneration of Chinese fir embryonic tissue based on immature zygotic embryos [D].
  • Somatic embryogenesis belongs to asexual reproduction technology.
  • the seedlings produced have little genetic background differences and the seedlings are neat. Compared with cuttings and traditional tissue culture, its scale is larger and the cost is lower.
  • somatic embryogenesis and angiosperm somatic embryogenesis have completely different pathways and mechanisms. It is very difficult for mature seeds and plants developed from seeds to transform vegetative cells into embryogenic cells.
  • some of the nucellar cells are in the undifferentiated stage during the development of the female gametophyte into seeds after fertilization. Under proper culture conditions, the characteristics of fission and polyembryonic can be used to induce the formation of somatic embryos. Gupta et al.
  • the technical problem to be solved by the present invention is to provide a method for promoting Chinese fir somatic embryogenesis by using salicylic acid, which can improve the induction efficiency of Chinese fir somatic embryos.
  • a method of using salicylic acid to promote Chinese fir somatic embryogenesis Take Chinese fir embryonic embryonic stalk mass and add it to the somatic embryo induction medium, and conduct dark culture in an incubator to induce and promote Chinese fir somatic embryogenesis; among them, somatic embryo induction culture Base: DCR is used as the basic medium, supplemented with salicylic acid at a concentration of not more than 8mg/L.
  • the somatic embryo induction medium also includes: 1mg/L VC, 0.45g/L glutamine, 0.5g/L hydrolyzed casein, 2 ⁇ 4g/L Ac, 2.8g/L crystal agar, 0.2g/L L asparagine, 0.2g/L proline, 100 ⁇ 200g/L PEG, 1 ⁇ 5mg/L GA, 2 ⁇ 8mg/L ABA.
  • the concentration of the salicylic acid is 2-8 mg/L.
  • the concentration of the salicylic acid is 6 mg/L.
  • the culture temperature of the dark culture is 23°C, and the culture time is 60-100 days.
  • the concentration of salicylic acid is 6-8 mg/L.
  • the method for obtaining the embryonic stalk mass of Chinese fir is: taking the immature cones of Chinese fir to obtain the seeds of fir, sterilizing the seeds of the fir in an ultra-clean workbench, removing the seed coat under a microscope, cutting the endosperm to take out the seed embryo, Put the seed embryos in a callus induction medium and culture in the dark at a culture temperature of 23°C for 25-30 days to obtain a Chinese fir embryonic stalk mass; wherein the callus induction medium is DCR medium Add 0.5mg/LKT, 0.5mg/L BA, 1mg/L VC, 1 ⁇ 2mg/L 2,4-D, 0.45g/L glutamine, 0.5g/L hydrolyzed casein, 2.5g/L Ac , 2.3g/L crystal seaweed.
  • the advantages of the present invention are: the method of using salicylic acid to promote somatic embryogenesis of Chinese fir provided by the present invention uses DCR as the basic medium with additional concentration of no more than 8mg/L salicylic acid. Acid, take the Chinese fir embryonic embryonic stalk mass and inoculate it on the somatic embryo induction medium, and conduct dark culture in an incubator at a temperature of 23 degrees to induce and promote Chinese fir somatic embryogenesis. Cultivate for 60-100 days.
  • Figure 1 is a statistical diagram of the number of embryogenesis in mature cotyledons of Chinese fir A treated with different salicylic acid concentrations;
  • Figure 2 shows the developmental status of Chinese fir A somatic embryos under different culture times and different concentrations of salicylic acid under stereomicroscope; a to d are embryonic embryonic stalks supplemented with 6 mg/L salicylic acid at 0, 10, 20, and 30 days Group development process, e ⁇ i are respectively the development of embryogenic embryonic stalk group with 0, 2, 4, 6, 8 mg/L salicylic acid added at 45 days, scale a ⁇ d is 2 mm, scale e ⁇ f is 5 mm ;
  • Figure 3 is a statistical chart showing the occurrence of mature cotyledon embryo B under different concentrations of salicylic acid
  • Figure 4 shows the developmental status of Chinese fir somatic embryo B under different culture time and different concentration of salicylic acid under stereo microscope;
  • Figures a, b, c, and d represent 0d, 10d, 20d, and 30d, respectively, with 8mg/L salicylic acid added
  • the process of somatic embryogenesis; e, f, g, h, i represent the occurrence of somatic embryo B of fir under the treatment of 0, 2, 4, 6, 8 mg/L salicylic acid at 45 days, and the scales a ⁇ d are 2mm, and e ⁇ The f scale is 5mm.
  • the raw materials used in the following examples are immature cones randomly collected from Yangkou Forest Farm in Fujian, and marked, and the embryogenic stalk masses induced by immature zygotic embryos (labeled A, B, randomly from two different Chinese fir) is the starting material.
  • Salicylic acid is easily soluble in water. Weigh 1.0g of salicylic acid powder in a beaker and dissolve it with ultrapure water, and dilute to 1L, prepare 1mg/mL salicylic acid mother liquor, and finally transfer it to Store in a brown bottle in a refrigerator at 4°C for later use.
  • Embryogenic stalk mass induction and proliferation medium Chinese fir embryonic stalk mass induction is the process of inducing callus formation from Chinese fir seeds.
  • the medium formula is DCR medium with 0.5mg/L KT, 0.5mg/L BA, 1mg /LVC, 1 ⁇ 2mg/L 2,4-D, 0.45g/L glutamine, 0.5g/L hydrolyzed casein, 2.5g/L Ac, 2.3g/L crystal agar.
  • Somatic embryo induction medium Chinese fir somatic embryo induction is the process of inducing Chinese fir callus to form somatic embryo.
  • the medium formula is DCR basic medium with 1mg/L VC, 0.45g/L glutamine, 0.5g/L hydrolyzed casein, 2 ⁇ 4g/L Ac, 2.8g/L crystal agar, 0.2g/L Asparagine, 0.2g/L proline. 100 ⁇ 200g/L PEG, 1 ⁇ 5mg/L GA, 2 ⁇ 8mg/L ABA.
  • Somatic embryo induction The embryonic embryo stalk masses A and B were added to different concentrations (0, 2, 4, 6, 8 mg/L) of salicylic acid somatic embryo medium, so as not to add exogenous salicylic acid (0 mg/L). /L) The somatic embryo material induced by the culture medium is used as a control. Each dish is inoculated with 6 calli, at least 3 dishes are repeated, cultured in dark, and the culture temperature is 23 degrees.
  • each concentration (0, 2, 4, 6, 8 mg/L) produced cotyledon embryos, and the average number of mature cotyledon embryos per dish of SA with 0 ⁇ 8 mg/L was 6, in order. 8, 10.3, 13.3, 10.3; when the concentration is 6mg/L, the number of somatic embryos is the largest, which is about 2.21 times that of the control group, which is significantly different from the control group; the content of somatic embryos per dish at the other concentrations is higher than that of the control group, but there is no Significant difference; at 100 days, the number of mature cotyledon embryos of each concentration gradually increased and reached the maximum.
  • the average number of mature cotyledon embryos per dish in the control group was 18; the number of somatic embryos at 6 mg/L was about 36.3, which was about
  • the control group is 2.01 times that of the control group, which is significantly different from the control group; the number of cotyledon embryos at other concentrations has no significant difference compared with the control group, indicating that 6mg/L SA contributes to the embryogenic embryonic stalk mass A somatic embryogenesis.
  • each concentration of 0-8mg/LSA has mature cotyledon embryos.
  • the concentration increases, the number of mature cotyledon embryos gradually increases; when the SA concentration is 8mg/L, mature cotyledons The number of embryos is the largest, which is a significant difference from the control group.
  • the cotyledon embryos are all mature and the number of mature cotyledon embryos increases.
  • the SA concentration is 8mg/L, the number of cotyledon embryos is the largest, which is mature per dish on average.
  • the number of cotyledon embryos is 60.3, and the average number of cotyledon embryos per dish in the control group is 30.3, which is 1.99 times that of the control group.
  • the somatic embryogenesis promotion effect of stalk mass B is the best. It can be seen from Figure 4 that at 0d, the embryonic stalk mass is transparent (Figure 4a); at 10d, the embryonic head becomes dense and dense ( Figure 4b); at 20d, the embryonic stalk becomes green, and the hypocotyl becomes longer and changed. Red ( Figure 4c) transforms into columnar embryos; at 30d, columnar embryos are produced, and the stalk continues to elongate and become thinner ( Figure 4d). Compared with the control group, the content of columnar embryos is higher (Figure 4d); at 45d, it is different Concentrations of salicylic acid treatment of hypoembryonic stalk mass B produced cotyledon embryos ( Figure 4e-4i).

Abstract

本发明公开了一种利用水杨酸促进杉木体胚发生的方法，属于植物组培技术领域。本发明所提供的方法具体为：以DCR为基本培养基，附加有浓度不超过8mg/L的水杨酸作为杉木体胚诱导培养基；通过杉木未成熟球果获得未成熟种胚，将其放在在愈伤诱导培养基上培养25～30天获得杉木胚性胚柄团；将所得杉木胚性胚柄团放入杉木体胚诱导培养基中，于培养箱中进行暗培养，诱导促进杉木体胚发生，23℃条件下，培养60～100天，即可得到比对照数目最高可达2倍以上的体胚，大大提高了杉木体胚的诱导效率，具有很好的实用性。

Description

一种利用水杨酸促进杉木体胚发生的方法 技术领域
本发明属于植物组培技术领域，更具体地说，涉及一种利用水杨酸促进杉木体胚发生的方法。
背景技术
杉木(Cunninghina lanceolata)隶属于杉科杉木属，是我国重要用材树种之一，其生长快、干形通直圆满，材质轻软细致，易加工，纹理美观，有特殊香气，不翘不裂，抗蛀耐腐，是建筑、家具、桥梁、造船、造纸及木纤维等工业的优良原材料。目前，主要通过三种途径繁殖杉木种苗：种子、扦插和传统组织培养[温亚锋.基于未成熟合子胚的杉木胚性组织诱导和继代的研究[D].福建农林大学，2011.]，但是良种种子产量、种子播种品质不稳定，第2代以上高级良种供应不足，且种子播种品质差，尤其是种子涩粒多，通常占重量的40％～60％[郑仁华.杉木遗传育种研究进展与对策[J].世界林业研究，2005(03)：63-65.]；而扦插育苗则受到母树穗条的多少、场地空间及生根率等方面的限制，其规模很难满足现代林业生产对杉木种苗的大量需求；此外，传统的组织培养根的诱导较为困难，诱导出的根数量少且弱，在一定程度上影响了苗木质量和造林的成功率。因此建立高效率的杉木未成熟胚的胚胎发生体系，有利于加快杉木速生优苗的产业化发展。
体细胞胚发生属于无性繁殖技术，生产出的种苗遗传背景差异小，苗木整齐。与扦插和传统组织培养相比，其规模更大、成本更低。然而，作为裸子植物的杉木，体胚的发生与被子植物的体细胞胚发生有着完全不同的途径和机理，成熟的种子以及由种子发育而来的植株，营养细胞转变为胚性细胞十分困难。但在针叶树种中，雌配子体受精后发育成种子的过程中有一部分珠心细胞处于未分化的阶段，在适当的培养条件下，可以利用裂生多胚特性，诱导体胚形成。Gupta等(1985，1995)报道过利用这一生物学特性，诱导出花旗松(Pseudotsuga menziesii)和湿地松(Pinuselliottii)的体胚，其大致可以分为四个步骤：胚性胚柄细胞团(ESM：Embryogenesissuspensor mass)的诱导、保持和增殖、体胚的成熟、萌发和植株再生，并证明该技术体系具有达到产业化的前景。
然而，目前虽有采用杉木未成熟胚诱导体胚发生的报道，但是只能在外植体上形成个别体细胞胚，存在诱导率低、数量少的问题，难以在生产上应用。因此建立高效率的杉木未成熟胚的胚胎发生体系，有利于加快杉木速生优苗的产业化发展。已有研究表明水杨酸对不同植物的体胚发生具有不同的效果，其中的机理还待进一步探索，但目前未见将其应用于杉木体胚发生的报导。
发明内容
针对现有技术存在的上述问题，本发明所要解决的技术问题在于提供一种利用水杨酸促进杉木体胚发生的方法，能够提高杉木体胚诱导效率。
为了解决上述技术问题，本发明所采用的技术方案如下：
一种利用水杨酸促进杉木体胚发生的方法：取杉木胚性胚柄团加入体胚诱导培养基中，于培养箱中进行暗培养，诱导促进杉木体胚发生；其中，体胚诱导培养基：以DCR为基本培养基，附加有浓度不超过8mg/L的水杨酸。
所述的体胚诱导培养基还包括：1mg/L VC，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2～4g/L Ac，2.8g/L水晶洋菜，0.2g/L天冬酰胺，0.2g/L脯氨酸，100～200g/L PEG，1～5mg/L GA，2～8mg/L ABA。
所述的水杨酸，其浓度为2～8mg/L。
所述的水杨酸，其浓度为6mg/L。
所述暗培养的培养温度为23℃，培养时间为60～100天。
所述的水杨酸浓度为6-8mg/L。
所述杉木胚性胚柄团的获取方法为：取杉木未成熟球果得到杉木种子，在超净工作台中让杉木种子先灭菌后，在显微镜下去除种皮，切开胚乳取出种胚，将种胚放入愈伤诱导培养基中，暗培养，培养温度为23度，培养25～30天，即得杉木胚性胚柄团；其中，所述的愈伤诱导培养基为DCR培养基中加入0.5mg/LKT，0.5mg/L BA，1mg/L VC，1～2mg/L 2,4-D，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2.5g/L Ac，2.3g/L水晶洋菜。
有益效果：相比于现有技术，本发明的优点为：本发明所提供的利用水杨酸促进杉木体胚发生方法，是以DCR为基本培养基，附加有浓度不超过8mg/L水杨酸，取杉木胚性胚柄团接种到所述的体胚诱导培养基上，于培养箱中进行暗培养，培养温度为23度，诱导促进杉木体胚发生，培养60～100天，即可得到比对照数目最高可达2倍以上的体胚，大大提高了杉木体胚的诱导效率，由此建立了 高效率的杉木未成熟胚的胚胎发生体系，有利于加快杉木速生优苗的产业化发展，具有很好的实用性。
附图说明
图1为不同浓度水杨酸处理下杉木A成熟子叶胚发生数统计图；
图2为体视镜下不同培养时间不同浓度水杨酸处理下杉木A体胚发育状态图；a～d是添加6mg/L水杨酸在0、10、20、30天的胚性胚柄团发育过程，e～i分别是45d时添加0、2、4、6、8mg/L的水杨酸胚性胚柄团发育情况，a～d标尺为2毫米，e～f标尺为5毫米；
图3为不同浓度水杨酸处理下成熟子叶胚B发生数统计图；
图4为体视镜下不同培养时间不同浓度水杨酸处理下杉木体胚B发育状态图；图a、b、c、d分别代表0d、10d、20d、30d加入8mg/L水杨酸的体胚发生过程；e、f、g、h、i分别代表45d时0、2、4、6、8mg/L水杨酸处理下杉木体胚B发生情况，a～d标尺为2mm，e～f标尺为5mm。
具体实施方式
下面结合具体实施例对本发明进一步进行描述。
以下实施例中所使用的原材料为福建洋口林场随机采集的未成熟球果，并做好标记，通过未成熟合子胚诱导的胚性胚柄团(标记为A、B，随机来自不同的两棵杉木)为起始材料。
实施例1
母液配制：水杨酸易溶于水，称取1.0g水杨酸粉末于烧杯中用超纯水溶解，并定容至1L，配置成1mg/mL的水杨酸母液，最后将其转移到4℃冰箱中，用棕瓶保存备用。
胚性胚柄团诱导和增殖培养基：杉木胚性胚柄团诱导是杉木种子诱导形成愈伤的过程，培养基配方为DCR培养基中加入0.5mg/L KT，0.5mg/L BA，1mg/LVC，1～2mg/L 2,4-D，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2.5g/L Ac，2.3g/L水晶洋菜。
体胚诱导培养基：杉木体胚诱导是杉木愈伤诱导形成体胚的过程。培养基配方为DCR基本培养基中加入1mg/L VC，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2～4g/L Ac，2.8g/L水晶洋菜，0.2g/L天冬酰胺，0.2g/L脯氨酸。100～200g/L PEG，1～5mg/L GA，2～8mg/L ABA。
胚性胚柄团诱导与增殖：在超净工作台中让杉木种子先灭菌后，在显微镜下去除种皮，切开胚乳取出种胚，将胚放入愈伤诱导培养基中，胚和培养基要尽可能接触，这样有助于愈伤组织的形成。杉木愈伤继代周期为25～30天，暗培养，培养温度为23度。
体胚诱导：将胚性胚柄团A和B分别加入不同浓度(0、2、4、6、8mg/L)的水杨酸体胚培养基中，以不添加外源水杨酸(0mg/L)培养基诱导的体胚材料为对照，每皿接种6块愈伤，至少3皿重复，暗培养，培养温度为23度。
采用One Way ANOVA、多重比较检验法进行数据统计和分析，分析软件为SPSS 16.0，显著性水平设为α＝0.05，采用GraphPad对SPSS软件分析结果进行绘图。
(1)胚性胚柄团A的体胚发生
如图1可知，60d时，每个浓度(0、2、4、6、8mg/L)均有子叶胚产生，0～8mg/L的SA的平均每皿成熟子叶胚个数依次为6、8、10.3、13.3、10.3；浓度为6mg/L时体胚个数最多，约为对照组的2.21倍，与对照组差异显著；其余浓度每皿的体胚含量都比对照组高，但没有显著差异；在100d时，各个浓度的成熟子叶胚的数量逐渐增多并达到最大值，对照组每皿的成熟子叶胚平均数量为18个；6mg/L的体胚个数约36.3个，约为对照组的2.01倍，与对照组差异显著；其余浓度下子叶胚数量与对照组相比均无显著差异，说明6mg/L的SA有助于胚性胚柄团A体胚发生。由图2可知，在30d时，6mg/L水杨酸处理下已经产生了子叶胚(图2d)，而对照组还处于柱状胚时期(图2d)，说明水杨酸能促进体胚生长；在45d时，0～8mg/L水杨酸处理下均产生了子叶胚。综上所诉，6mg/L水杨酸能促进胚性胚柄团A体胚发生。
(2)胚性胚柄团B的体胚发生
由图3可知，60d时，0～8mg/LSA每个浓度均有成熟子叶胚产生，随着浓度的升高，成熟子叶胚的个数逐渐增加；其中SA浓度在8mg/L时，成熟子叶胚的个数最多，与对照组形成了显著差异；在100d时，子叶胚均发育成熟，且成熟子叶胚个数增加，SA浓度为8mg/L时，子叶胚个数最多，平均每皿成熟子叶胚个数为60.3，对照组平均每皿的子叶胚个数为30.3，是对照组的1.99倍，说明在0～8mg/L的SA浓度中，8mg/L的水杨酸对胚性胚柄团B的体胚发生促进效果最佳。由图4可知，在0d时，胚性胚柄团颜色偏透明(图4a)；10d时，胚头变 的变密致密(图4b)；20d时，胚头变绿，胚轴延长且变红(图4c)向柱状胚转变；30d时，柱状胚产生，胚柄继续伸长且变细(图4d)，与对照组相比，柱状胚含量较多(图4d)；45d时，不同浓度水杨酸处理下胚性胚柄团B均产生了子叶胚(图4e～4i)。

Claims (6)

  1. 一种利用水杨酸促进杉木体胚发生的方法，其特征在于，取杉木胚性胚柄团加入体胚诱导培养基中，于培养箱中进行暗培养，诱导促进杉木体胚发生；其中，体胚诱导培养基：以DCR为基本培养基，附加有浓度不超过8mg/L的水杨酸。
  2. 根据权利要求1所述的利用水杨酸促进杉木体胚发生的方法，其特征在于，所述的体胚诱导培养基还包括：1mg/L VC，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2～4g/L Ac，2.8g/L水晶洋菜，0.2g/L天冬酰胺，0.2g/L脯氨酸，100～200g/L PEG，1～5mg/L GA，2～8mg/L ABA。
  3. 根据权利要求2所述的利用水杨酸促进杉木体胚发生的方法，其特征在于，所述的水杨酸的浓度为2～8mg/L。
  4. 根据权利要求1～3任一所述的利用水杨酸促进杉木体胚发生的方法，其特征在于，所述暗培养的培养温度为23℃，培养时间为60～100天。
  5. 根据权利要求1～3任一所述的利用水杨酸促进杉木体胚发生的方法，其特征在于，所述的水杨酸浓度为6-8mg/L。
  6. 根据权利要求1～3任一所述的利用水杨酸促进杉木体胚发生的方法，其特征在于，所述杉木胚性胚柄团的获取方法为：取杉木未成熟球果得到杉木种子，在超净工作台中让杉木种子先灭菌后，在显微镜下去除种皮，切开胚乳取出种胚，将种胚放入愈伤诱导培养基中，暗培养，培养温度为23度，培养25～30天，即得杉木胚性胚柄团；其中，所述的愈伤诱导培养基为DCR培养基中加入0.5mg/L KT，0.5mg/L BA，1mg/L VC，1～2mg/L 2,4-D，0.45g/L谷氨酰胺，0.5g/L水解酪蛋白，2.5g/L Ac，2.3g/L水晶洋菜。
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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4197318A1 (en) 2021-12-17 2023-06-21 Albert-Ludwigs-Universität Freiburg Phosphodiesterases inhibitors to promote in vitro plant cell reprogramming towards plant embryogenesis or microcallus formation

Families Citing this family (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN115735769B (zh) * 2022-11-25 2023-09-08 南京林业大学 一种通过控制氧化还原稳态来促进杉木体胚发生的方法

Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101130763A (zh) * 2007-08-08 2008-02-27 南京林业大学 银杏愈伤组织或悬浮细胞的快速繁殖方法
CN106538394A (zh) * 2016-12-20 2017-03-29 江西宜信堂医疗科技有限公司 一种红豆杉培养基及其制备方法
CN106688895A (zh) * 2017-03-27 2017-05-24 南京林业大学 一种利用水杨酸进行杂交鹅掌楸体胚发生的方法

Family Cites Families (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101983556B (zh) * 2010-09-07 2011-11-30 南京林业大学 杉木体细胞胚胎发生及植株再生方法

Patent Citations (3)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101130763A (zh) * 2007-08-08 2008-02-27 南京林业大学 银杏愈伤组织或悬浮细胞的快速繁殖方法
CN106538394A (zh) * 2016-12-20 2017-03-29 江西宜信堂医疗科技有限公司 一种红豆杉培养基及其制备方法
CN106688895A (zh) * 2017-03-27 2017-05-24 南京林业大学 一种利用水杨酸进行杂交鹅掌楸体胚发生的方法

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP4197318A1 (en) 2021-12-17 2023-06-21 Albert-Ludwigs-Universität Freiburg Phosphodiesterases inhibitors to promote in vitro plant cell reprogramming towards plant embryogenesis or microcallus formation
WO2023110673A1 (en) 2021-12-17 2023-06-22 Consejo Superior De Investigaciones Cientificas (Csic) Phosphodiesterases inhibitors to promote in vitro plant cell reprogramming towards plant embryogenesis or microcallus formation

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