WO2020107888A1 - 一种五小叶槭组培苗瓶外生根方法 - Google Patents

一种五小叶槭组培苗瓶外生根方法 Download PDF

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WO2020107888A1
WO2020107888A1 PCT/CN2019/093768 CN2019093768W WO2020107888A1 WO 2020107888 A1 WO2020107888 A1 WO 2020107888A1 CN 2019093768 W CN2019093768 W CN 2019093768W WO 2020107888 A1 WO2020107888 A1 WO 2020107888A1
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tissue culture
rooting
culture seedlings
seedlings
acer
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PCT/CN2019/093768
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French (fr)
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李飞鸿
谢松林
马建华
丁龙梅
朱晓菲
何程相
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四川七彩林科股份有限公司
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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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  • the invention belongs to the technical field of plant propagation, and in particular relates to a method for rooting out of a five-leaf maple tissue culture seedling.
  • maple plants There are about 200 species of maple plants in the world, mainly distributed in the northern temperate zone. The main producing areas are China and Japan. There are more than 150 species in China. They are widely distributed in the north and south provinces, but the distribution center is in the middle or west. Some maple trees are good for wood. Some trees are good nectar plants, and some can be used as shade trees and ornamental trees. The five-leaf maple is a rare ornamental tree. Up to now, there are only over 500 plants in the wild known as Acer palmatum, which belongs to the extremely small population and has been included in the "critically dangerous" grade in the "Chinese Biodiversity Red List ⁇ Higher Plant Volume". In February 2016, the Sichuan Provincial People's Government announced a list of 18 key protected wild plants in Sichuan province, and five small leaf maples were included.
  • Acer Pentaphyllum is a deciduous tree of the genus Acer of the Aceraceae family. It can grow up to 10 meters high, but most often grow up to four or five meters high and grow in river valleys with an altitude of 2200-3000m.
  • the five-leaf maple is the only species in the five-leaf maple family.
  • the five-leaf maple leaves change from green to yellow first, and finally to red. Its unique claw-like leaf shape and the colorful and changeable colors in autumn make it the object of attention of botanists and horticulturalists around the world. It is "the two largest ornamental maple trees in the world juxtaposed with red maple".
  • a cultivation method for the rapid propagation of five-leaf maple branches with the application number 201711422694.8 includes the following steps: Step one, collecting five years old and young shoots with a length of 4-6 cm The shoots with buds of Acer lobata are explant materials; in step two, the leaves with buds are cut off, cut into stalks with buds, and the stalks with buds are disinfected; in step three, the stalks with buds are cultured in medium, Medium MS+NAA0.05mg/L+6-BA0.05mg/L+GA3 2.0mg/L+AC 1.0g/L; Step four, strong seedling culture, the medium is MS+GA31.0mg/L+NAA0.
  • Step five rooting culture, the medium is White+IBA1.2mg/L+6-BA1.0mg/L+AC1.0g/L;
  • step Sixth 7 ⁇ 15d after rooting culture, move the tissue culture pot containing the five-leaf maple seedlings to outdoor shade shed or greenhouse for closed pot refining for 15 ⁇ 20d, the shading degree is 50% ⁇ 70%;
  • After finishing the seedlings open the tissue culture pot lid and open the seedlings under natural light for 3-7 days; after the seedlings are finished, use tweezers to remove the five-leaf maple seedlings from the tissue culture pot and wash the roots of the five-leaf maple seedlings
  • Step 7 Transplant directly, or use 0.1%-0.3% potassium permanganate solution or 500 times polydactyl solution to clean the five-leaf maple seedlings;
  • Step 8 Fertilize regularly after transplanting, conduct water and fertilizer management, and cultivate five Small leaf maple
  • the purpose of the present invention is to overcome the above-mentioned shortcomings of the prior art, and to provide a method for rooting outside the bottle of tissue culture seedlings of A. pentaphyllum with short seedling formation time and high rooting rate.
  • the present invention adopts the following technical solutions:
  • a method for rooting out of a bottle of tissue culture seedlings of Acer palmatum including the following steps:
  • Step 1 Prepare the tissue culture seedlings: Select the tissue culture seedlings of Acer pentaphyllum with a growth of 40 days and a height of 2 to 4 cm that are in good condition as the rooting material of the bottle, and cut the middle and lower leaves of the tissue culture seedlings;
  • Step 2 Rooting induction of tissue culture seedlings: Inoculate the tissue culture seedlings prepared in Step 1 in an induction medium for 2 to 10 days of induction culture.
  • the induction medium is 1/2MS+5-15mg/L IBA+25g/L Sucrose + 5g/L agar;
  • Step 3 Prepare the substrate: After sterilizing the mixed substrate of vermiculite and perlite by 800 times of carbendazim, put it into the bottom vent hole tray, the amount of the loaded substrate is 3/4 of the height of the hole tray, then pour water, then Watering by immersion pot every 2-3 weeks;
  • Step 4 Transplanting: Take the tissue cultured seedlings from the induction culture of Step 2, wash the base, insert it into the substrate prepared in Step 3 and compact it, one plant per hole, insert depth about 1cm, spray water on the leaf surface once, cover the hole The disc cover is sprayed with water twice within the first 1 to 5d, once a week for the 5th to 20d, and uncovered on the 25th for normal maintenance.
  • the induction medium is 1/2MS+10mg/L IBA+25g/L sucrose+5g/L agar.
  • the mass ratio of vermiculite and perlite is 1 to 3:1.
  • the mass ratio of vermiculite and perlite is 3:1.
  • step 2 is 2d.
  • tissue culture seedlings transplanted in step 4 are placed in an environment with a temperature of 22-27°C and a humidity of 80% or more.
  • the high concentration of inorganic salts in the MS medium can meet the requirements of rapid growth and accelerate the growth of callus and culture. In most plants, it reduces a large number of elements in the medium Concentration can improve the rooting ability of test-tube seedlings.
  • the 1/2MS basic medium described in this study is the MS basic medium with a large number of elements halved, other components unchanged, with low inorganic salt concentration, meet the needs of growth and help rooting.
  • the IBA is indobutyric acid, a plant growth hormone, mainly used for cutting rooting, can induce the formation of root protoplasts, promote cell differentiation and division, is conducive to the formation of new roots and the differentiation of the vascular bundle system, promotes the insertion The formation of adventitious roots.
  • the rooting of the tissue culture seedlings of Acer pentaphyllum according to the present invention is affected by the concentration of exogenous hormones, rooting induction time and physiological state of tissue culture seedlings.
  • the present invention uses IBA as an exogenous hormone to indirectly induce adventitious roots by changing the content of endogenous hormones Occurrence and differentiation.
  • the tissue culture seedlings of Acer pentaphyllum after multiplication and propagation are inoculated into the induction medium for a period of time, and then transplanted into the substrate to induce rooting and domestication, so that the tissue culture seedlings gradually adapt to the external environment and improve the rooting rate and migration Survival rate after planting.
  • the transplanting and rooting process in the present invention is an acclimation process, which shortens the seedling formation time.
  • the present invention eliminates the root washing process in the existing tissue culture seedling transplanting method after rooting, and avoids the damage caused by root washing and pollution caused by inadequate cleaning; the method and process of rooting outside the bottle of the five-leaf maple tissue culture seedling of the present invention It is simple, easy to operate, and has a high rooting rate, which improves the survival rate of Acer palmatum.
  • FIG. 1 is a tissue culture seedling of a method of rooting out of a bottle of a five-leaf maple tissue culture seedling of the present invention
  • FIG. 2 is a rooting induction process of tissue culture seedlings of a method of rooting out of a bottle of tissue culture seedlings of Acer pentaphyllum according to the present invention
  • Fig. 3 is a rooting seedling after transplanting for 40 days in a method of rooting out of a bottle of tissue culture seedlings of Acer pentaphyllum according to the present invention.
  • a method for rooting out of a bottle of tissue culture seedlings of Acer palmatum including the following steps:
  • Step 1 Prepare the tissue culture seedlings: Select the tissue culture seedlings of Acer pentaphyllum with a growth of 40 days and a height of 2 to 4 cm that are in good condition as the rooting material of the bottle, and cut the middle and lower leaves of the tissue culture seedlings;
  • Step 2 Rooting induction of tissue culture seedlings: Inoculate the tissue culture seedlings prepared in Step 1 in an induction medium for 2 days of induction culture.
  • the induction medium is 1/2MS+10mg/L IBA+25g/L sucrose+5g/ L agar
  • Step 3 Prepare the matrix: mix vermiculite and perlite at a mass ratio of 3:1, sterilize by 800 times carbendazim, and put it into the bottom vent hole tray.
  • the amount of the matrix is 3/h of the height of the tray 4. Then pour through the water, use the immersion pot method for watering once on the 40th day, and use the immersion pot method for watering once on the 60th day;
  • Step 4 Transplanting: Take the tissue cultured seedlings from the induction culture of Step 2, wash the base, insert it into the substrate prepared in Step 3 and compact it, one plant per hole, insert depth about 1cm, spray water on the leaf surface once, cover the hole Dish cover, transplanted tissue culture seedlings are placed in an environment with a temperature of 22 to 27°C and a humidity of more than 80%. Spray water twice within the first 1 to 5d, spray water once a week from the 5th to 20d, and remove the cover on the 25th , Normal maintenance.
  • Example 2 The effect of different hormone concentration inducing media on rooting of tissue culture seedlings of Acer palmatum
  • Example 1 For the method of rooting outside the bottle of the tissue culture seedlings of Acer palmatum, see Example 1, set the concentration of the hormone IBA in the induction medium in Step 2 to 0mg/L, 5mg/L, 10mg/L, 15mg/L, and induce culture for 4 days The remaining steps are the same. Step 4 Rooting rate, indefinite root number and root length were counted on the 40th day after transplanting. The results are shown in Table 1.
  • Rooting rate (%) (number of roots of tissue culture seedlings / total number of tissue culture seedlings) ⁇ 100%;
  • Root length record the maximum indefinite root length of each tissue culture seedling as the root length of the tissue culture seedling
  • Table 1 The effect of different hormone concentration inducing media on rooting of A. pentaphyllum tissue culture seedlings
  • Example 2 For the method of rooting out of the bottle of tissue culture seedlings of A. pentaphyllum, see Example 1. Set the induction culture days in step 2 to 0d, 2d, 4d, 6d, 8d, and 10d, and the rest of the steps are the same. Step 4 The rooting rate, indefinite root number and root length were counted on the 40th day after transplanting. The results are shown in Table 2.
  • Rooting rate% Indefinite number (article) Root length cm 0d(CK) 53.33bc 2.26a 3.36a 2d 95.83a 3.33a 4.82a 4d 91.67a 3a 4.37a 6d 66.67b 2.75a 3.87a 8d 41.67c 2.5a 4.7a 10d 58.33bc 3a 3.5a
  • Step 4 Rooting rate, indefinite root number and root length were counted on the 40th day after transplanting. The results are shown in Table 3.
  • the substrate is made of vermiculite or perlite alone, and the rooting effect of A. pentaphyllum is not good.
  • the substrate with the mass ratio of vermiculite and perlite of 3:1 is the best, and the rooting rate reaches 95.83%
  • the number of adventitious roots is 3, and the root length is 4.82 cm, but the rooting situation is not significantly different from that of the five-leaf maple with a substrate with a mass ratio of 1:1 of vermiculite and perlite.
  • Example 5 Effect of tissue culture seedlings of Acer pentaphyllum at different heights on rooting of tissue culture seedlings of Acer pentaphyllum
  • Example 1 For the method of rooting outside the bottle of the tissue culture seedlings of Acer pentaphyllum, see Example 1.
  • the rooting material outside the bottle is 1.5cm, 2.5cm, 3.5cm, 4.5cm.
  • Step 4 The rooting rate, indefinite root number and root length were counted on the 40th day after transplanting. The results are shown in Table 4.

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Abstract

本发明提供了一种五小叶槭组培苗瓶外生根方法，包括以下步骤：选取增殖40d、高2～4cm生长状况良好的五小叶槭组培苗作为瓶外生根材料；将组培苗接种于1/2MS+10mg/L IBA+25g/L蔗糖+5g/L琼脂的诱导培养基中，诱导培养2～10d；将蛭石和珍珠岩混合并经多菌灵消毒后，装入底部透气穴盘中；取诱导培养后的组培苗，清洗基部，插入基质中压紧，每穴一株，插入深度约1cm，叶面喷水，盖上穴盘盖，定期喷水，第25d揭盖，正常养护。

Description

一种五小叶槭组培苗瓶外生根方法 技术领域
本发明属于植物繁殖技术领域，具体涉及一种五小叶槭组培苗瓶外生根方法。
背景技术
全球槭树科植物约200种，主要分布于北温带，主产地为我国和日本，我国有150种以上，广布于南北各省，但分布中心为中部或西部，有些槭树是良好的木材用树，有些是很好的蜜源植物，有些可作为荫蔽树和观赏树。五小叶槭就是稀罕的观赏树。至目前为止，已知五小叶槭野外现仅存500余株，五小叶槭属于极小种群，在《中国生物多样性红色名录·高等植物卷》里已列入“极危”等级。2016年2月，四川省人民政府公布了18种四川省重点保护野生植物名录，五小叶槭被列入其中。
五小叶槭(Acer Pentaphyllum)，是槭树科槭属落叶乔木，可长至10米高，但多长至四五米高，生长在海拔2200-3000m的河谷地带。在槭属中，五小叶槭是五小叶槭系唯一的种。秋天，五小叶槭叶片先是由绿变黄，最后变为红色，其独特的鸡爪状叶形和秋季绚丽多变的色彩，使它成为全球植物学家和园艺学家关注的对象，有人称它为“与红枫并列的世界两大观赏枫树”。
目前，五小叶槭的繁殖技术主要是播种繁殖，但是种子发芽缓慢且发芽率低。目前也有研究五小叶槭枝条快速繁殖方法，如申请号为201711422694.8的一种五小叶槭枝条快速繁殖的培养方法，包括以下步骤：步骤一，采集3年生以下，嫩梢长度为4～6cm的五小叶槭带芽枝条为外植体材料；步骤二，带芽枝条剪去叶片，切割成带芽茎段，对带芽茎段进行消毒；步骤三，带芽茎段置于培养基中培养，培养基MS+NAA0.05mg/L+6-BA0.05mg/L+GA3 2.0mg/L+AC 1.0g/L；步骤四，壮苗培养，培养基为MS+GA31.0mg/L+NAA0.05mg/L+6-BA0.05mg/L+AC 1.0g/L；步骤五，生根培养，培养基为White+IBA1.2mg/L+6-BA1.0mg/L+AC 1.0g/L；步骤六，生根培养后7～15d，将装有五小叶槭苗的组培罐移至室外遮阴棚或温室中进行闭罐炼苗15～20d，遮阴度为50％～70％；闭罐炼苗结束后，再打开组培罐盖，在自然光下进行开罐炼苗3～7d；炼苗结束后，用镊子将五小叶槭苗从组培罐中移出，洗净五小叶槭苗根部；步骤七，直接移栽，或使用0.1％～0.3％的高锰酸钾溶液或500倍多灵菌溶液清洗五小叶槭苗；步骤八，移栽后定期施肥，进行水肥管理，培育得到五小叶槭树苗。但是该方法需要壮苗培养、生根培养等过程，过程复杂，且繁殖周期长，并且步骤六炼苗后需要洗净五小叶槭苗根部，清洗过程会对苗造成一定 损伤，如若清洗不到位甚至容易污染死亡。因此，急需一种生根率高、成活率高且成苗时间短的五小叶槭繁殖方法。
发明内容
本发明目的在于克服上述现有技术的不足，提供一种成苗时间短、高生根率的五小叶槭组培苗瓶外生根方法。
为实现上述目的，本发明采用以下技术方案：
一种五小叶槭组培苗瓶外生根方法，包括以下步骤：
步骤一：准备组培苗：选取增殖40d、高2～4cm生长状况良好的五小叶槭组培苗作为瓶外生根材料，将组培苗中下部叶片剪去；
步骤二：组培苗生根诱导：将步骤一准备的组培苗接种于诱导培养基中，诱导培养2～10d，所述诱导培养基为1/2MS+5～15mg/L IBA+25g/L蔗糖+5g/L琼脂；
步骤三：准备基质：将蛭石和珍珠岩混合基质经800倍的多菌灵消毒后，装入底部透气穴盘中，装入基质的量为穴盘高度的3/4，然后浇透水，之后每隔2-3周采用浸盆法浇水1次；
步骤四：移栽：取步骤二的诱导培养后的组培苗，清洗基部，插入步骤三准备的基质中压紧，每穴一株，插入深度约1cm，叶面喷水一次，盖上穴盘盖，第1～5d内喷水2次，第5～20d一周喷水一次，第25d揭盖，正常养护。
进一步地，所述诱导培养基为1/2MS+10mg/L IBA+25g/L蔗糖+5g/L琼脂。
进一步地，所述蛭石和珍珠岩的质量比为1～3:1。
进一步地，所述蛭石和珍珠岩的质量比为3:1。
进一步地，步骤二所述诱导培养为2d。
进一步地，步骤四中移栽的组培苗放置在温度为22～27℃，湿度为80％以上的环境中。
所述MS培养基的无机盐浓度高，尤其是铵盐和硝酸盐含量大，能满足迅速增长，加快愈伤组织和培养物生长的作用，在大多数植物中，降低培养基中的大量元素浓度，可提高试管苗的生根能力，本研究所述的1/2MS基本培养基为MS基本培养基大量元素减半，其他成分不变，具有无机盐浓度低，满足生长需要，有利于生根。
所述IBA为吲哚丁酸，是一种植物生长激素，主要用于插条生根，可诱导根原体的形成，促进细胞分化和分裂，有利于新根生成和维管束系统的分化，促进插条不定根的形成。
本发明的五小叶槭组培苗生根情况受外源激素浓度、生根诱导时间和组培苗生理状 态等影响，本发明采用IBA作为外源激素，通过改变内源激素的含量来间接诱导不定根的发生和分化。高浓度外源激素促进组培苗生根和根系生长，但过高的浓度则抑制组培苗生根和根系生长；激素浓度低、生根诱导时间过短，则组培苗吸入少量外源激素，移栽后成活率低，而生根诱导时间过长，组培苗中外源激素含量过高，对组培苗的生根则起抑制作用；此外，同一植物不同生长状态对激素浓度的响应也存在差异。
相比现有技术，本发明的有益效果在于：
本发明将增殖扩繁后的五小叶槭组培苗，接种到诱导培养基一段时间后，移栽至基质中，进行诱导生根和驯化，使组培苗逐渐适应外界环境，提高生根率和移栽后的成活率。同时本发明中的移栽生根过程即为驯化过程，缩短了成苗时间。本发明省去现有的组培苗生根后移栽方法中的洗根过程，避免了洗根造成的损伤和清洗不到位造成污染；本发明的五小叶槭组培苗瓶外生根方法，过程简单，易于操作，且生根率高，提高了五小叶槭的成活率。
附图说明
图1是本发明一种五小叶槭组培苗瓶外生根方法的组培苗；
图2是本发明一种五小叶槭组培苗瓶外生根方法的组培苗生根诱导过程；
图3是本发明一种五小叶槭组培苗瓶外生根方法的移栽40d后的生根苗。
具体实施方式
以下实施例用于说明本发明，但不用来限定本发明的保护范围。若未特别指明，实施例中所用技术手段为本领域技术人员所熟知的常规手段。
实施例1
一种五小叶槭组培苗瓶外生根方法，包括以下步骤：
步骤一：准备组培苗：选取增殖40d、高2～4cm生长状况良好的五小叶槭组培苗作为瓶外生根材料，将组培苗中下部叶片剪去；
步骤二：组培苗生根诱导：将步骤一准备的组培苗接种于诱导培养基中，诱导培养2d，所述诱导培养基为1/2MS+10mg/L IBA+25g/L蔗糖+5g/L琼脂；
步骤三：准备基质：将蛭石和珍珠岩按质量比为3:1混合后经800倍的多菌灵消毒后，装入底部透气穴盘中，装入基质的量为穴盘高度的3/4，然后浇透水，第40d采用浸盆法浇水一次，第60d采用浸盆法浇水一次；
步骤四：移栽：取步骤二的诱导培养后的组培苗，清洗基部，插入步骤三准备的基质中压紧，每穴一株，插入深度约1cm，叶面喷水一次，盖上穴盘盖，移栽的组培苗放置在温度为22～27℃，湿度为80％以上的环境中，第1～5d内喷水2次，第5～20d一周喷水一次，第 25d揭盖，正常养护。
实施例2组培苗生根诱导时不同激素浓度的诱导培养基对五小叶槭组培苗生根的影响
五小叶槭组培苗瓶外生根方法参见实施例1，将步骤二中诱导培养基中的激素IBA的浓度设置为0mg/L、5mg/L、10mg/L、15mg/L，诱导培养4d，其余步骤相同。步骤四移栽后第40d统计生根率、不定根数和根长，结果见表1。
生根率(％)＝(组培苗生根株数/组培苗总株数)×100％；
根长：将每株组培苗最大不定根长度记为该株组培苗的根长；
表1不同激素浓度的诱导培养基对五小叶槭组培苗生根的影响
Figure PCTCN2019093768-appb-000001
由表1可知：试验以使用1/2MS+10mg/L IBA+25g/L蔗糖+5g/L琼脂的诱导培养基，诱导培养4d的五小叶槭生根效果最好，生根率达到91.67％，不定根数达3条，根长为4.37cm。
实施例3组培苗生根诱导的不同诱导天数对五小叶槭组培苗生根的影响
五小叶槭组培苗瓶外生根方法参见实施例1，将步骤二中诱导培养天数设置为0d、2d、4d、6d、8d、10d，其余步骤相同。步骤四移栽后第40d统计生根率、不定根数和根长，结果见表2。
表2不同诱导天数对五小叶槭组培苗生根的影响
诱导天数 生根率％ 不定根数(条) 根长cm
0d(CK) 53.33bc 2.26a 3.36a
2d 95.83a 3.33a 4.82a
4d 91.67a 3a 4.37a
6d 66.67b 2.75a 3.87a
8d 41.67c 2.5a 4.7a
10d 58.33bc 3a 3.5a
由表2可知：以生根诱导处理2d的五小叶槭效果最好，生根率达到95.83％，不定根数达3.33条，根长为4.82cm。
实施例4不同基质对五小叶槭组培苗生根的影响
五小叶槭组培苗瓶外生根方法参见实施例1，将步骤三的基质即蛭石和/或珍珠岩按质量比为 0:1，1:0，1:1，3:1设置四组，其余步骤相同。步骤四移栽后第40d统计生根率、不定根数和根长，结果见表3。
表3不同基质对五小叶槭组培苗生根的影响
Figure PCTCN2019093768-appb-000002
由表3可知：基质采用单独的蛭石或珍珠岩，五小叶槭生根效果不好，试验以蛭石和珍珠岩质量比为3:1的基质的五小叶槭效果最好，生根率达到95.83％，不定根数达3条，根长为4.82cm，但与以蛭石和珍珠岩质量比为1:1的基质的五小叶槭生根情况差异不显著。
实施例5不同高度的五小叶槭组培苗对五小叶槭组培苗生根的影响
五小叶槭组培苗瓶外生根方法参见实施例1，瓶外生根材料选取高为1.5cm，2.5cm，3.5cm，4.5cm生长状况良好的五小叶槭组培苗，其余步骤相同。步骤四移栽后第40d统计生根率、不定根数和根长，结果见表4。
表4不同高度的组培苗对五小叶槭组培苗生根的影响
Figure PCTCN2019093768-appb-000003
由表4可知：高为1.5cm的五小叶槭生根率达95.83％，但不定根数仅为1.85条。因此，试验以高为3.5cm的五小叶槭效果最好，生根率达到91.67％，不定根数达3.06条，根长为4.07cm。
以上所述之实施例，只是本发明的较佳实施例而已，仅仅用以解释本发明，并非限制本发明实施范围，对于本技术领域的技术人员来说，当然可根据本说明书中所公开的技术内容，通过置换或改变的方式轻易做出其它的实施方式，故凡在本发明的原理及工艺条件所做的变化和改进等，均应包括于本发明申请专利范围内。

Claims (6)

  1. 一种五小叶槭组培苗瓶外生根方法，其特征在于，包括以下步骤：
    步骤一：准备组培苗：选取增殖40d、高2～4cm生长状况良好的五小叶槭组培苗作为瓶外生根材料，将组培苗中下部叶片剪去；
    步骤二：组培苗生根诱导：将步骤一准备的组培苗接种于诱导培养基中，诱导培养2～10d，所述诱导培养基为1/2MS+5～15mg/L IBA+25g/L蔗糖+5g/L琼脂；
    步骤三：准备基质：将蛭石和珍珠岩混合基质经800倍的多菌灵消毒后，装入底部透气穴盘中，装入基质的量为穴盘高度的3/4，然后浇透水，之后每隔2～3周采用浸盆法浇水1次；
    步骤四：移栽：取步骤二的诱导培养后的组培苗，清洗基部，插入步骤三准备的基质中压紧，每穴一株，插入深度约1cm，叶面喷水一次，盖上穴盘盖，第1～5d内喷水2次，第5～20d一周喷水一次，第25d揭盖，正常养护。
  2. 根据权利要求1所述的一种五小叶槭组培苗瓶外生根方法，其特征在于，所述诱导培养基为1/2MS+10mg/L IBA+25g/L蔗糖+5g/L琼脂。
  3. 根据权利要求1所述的一种五小叶槭组培苗瓶外生根方法，其特征在于，所述蛭石和珍珠岩的质量比为1～3:1。
  4. 根据权利要求1所述的一种五小叶槭组培苗瓶外生根方法，其特征在于，所述蛭石和珍珠岩的质量比为3:1。
  5. 根据权利要求1所述的一种五小叶槭组培苗瓶外生根方法，其特征在于，步骤二所述诱导培养为2d。
  6. 根据权利要求1所述的一种五小叶槭组培苗瓶外生根方法，其特征在于，步骤四中移栽的组培苗放置在温度为22～27℃、湿度为80％以上的环境中。
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