香石竹的离体培养与试管开花研究
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摘要
利用组织培养的方法,研究香石竹(Dianthus caryophyllus L.)的离体培养、试管开花以及花芽诱导过程中其内部生理生化的变化,建立香石竹的快速繁殖体系,缩短营养生长向生殖生长转化的时间,探讨其花芽分化规律在理论上和应用上都具有重要的意义。
本文研究了不同浓度的6-BA、NAA,不同的琼脂浓度以及不同的封口材料对香石竹叶片和带有腋芽的茎段诱导的影响。研究发现,6-BA和NAA对香石竹叶片和腋芽的诱导率具有显著性差异。6-BA和NAA不同浓度的配比组合对叶片和腋芽诱导的影响也有很大程度的差别,当6-BA和NAA浓度分别为2.0mg.L~(-1)和0.5mg.L~(-1)的组合时,芽的诱导率最高达到11.2%和87.9%。培养基中不同的琼脂浓度对芽的诱导率形成显著性影响,当琼脂浓度为7g.L~(-1)时,芽的长势最好且发育出许多叶子。不同的封口材料对香石竹叶片和腋芽诱导效果显著,使用透气封口膜芽生长速度快,利于植株枝条和叶片的旺盛生长和形成。通过筛选,香石竹的腋芽在不同条件下的发芽率均要比叶片的发芽率高。
本文重点研究了生根、愈伤组织、N/P含量、蔗糖浓度、6-BA,NAA等对香石竹组培苗花芽发生和花发育的影响。研究发现,根对花芽形成有显著影响,生根明显抑制花芽诱导。是否分化愈伤组织对香石竹花芽诱导的影响不大。调整了MS培养基中的氮和磷含量,明显提高了花芽的诱导率,其中1N和5P的配比下花芽诱导率最高。在一定范围内培养基中蔗糖含量与花芽诱导率成正相关,但是过高的含量会产生玻璃化苗。6-BA(0.5mg.L~(-1))和NAA(0.1mg.L~(-1))配比时,花芽诱导率最高,达到72.3%。通过筛选,诱导香石竹花芽形成效果最好的培养方法是:在改良的MS基本培养基(1倍氮,5倍磷),蔗糖浓度提高到35g.L~(-1),并加入适宜浓度的6-BA(0.5mg.L~(-1))和NAA(0.1mg.L~(-1))可诱导香石竹试管苗在60天内开花。2个月内统计花芽诱导率为70%左右。
以诱导花芽形成效果最佳的最佳培养基作为处理组,同时设一对照组:MS,附加蔗糖30 g.L~(-1),琼脂浓度6.5g.L~(-1),pH5.8。研究香石竹花芽诱导过程中植株内部生理生化的变化和相关性。花芽诱导率高的处理,其可溶性蛋白质、可溶性糖、内源激素含量在整体上都明显高于花芽诱导率低的处理。
In vitro,in vitro flowering and internal variation of physiological and biochemical during flower bud differentiation for Dianthus caryophyllus L.through tissue culture, can build up Dianthus caryophyllus L.system of intermediate propagation,decrease the juvenile phases from a vegetative growth to a reproductive growth stage,inquiry into flower bud differentiation regulation is significance not only in theory but also in application.
The objective of this research is to study the effects of differ culture medium, 6-BA,NAA,differ Agar and closure film during the induction of lateral bud of Dianthus caryophyllus L..The results of our research show that the contrast of differ 6-BA and NAA concentration is very remarkable on the induction rate of lateral bud and leaf,also the effects of differ 6-BA and NAA concentration go together with ratio combination are very difference.When 6-BA and NAA are combined in level of 2.0 mg.L~(-1)and 0.5 mg.L~(-1),the induction rate of lateral bud is highest at 87.9%and 11.2%. The effect of differ Agar of culture medium is very remarkable on the induction of lateral bud and leaf of Dianthus caryophyllus L.As its Agar is 7g.L~(-1),the induction rate is highest,bud grow best and grow many leafs.The effect of differ closure film of culture medium is very remarkable on the induction of lateral bud and leaf of Dianthus caryophyllus L.Through using closure film,the effect on the induction of lateral bud of Dianthus caryophyllus L.also is significance,results shows that closure film can make bud grow rapidly and become a plant.
The mostly objective of this research is to study the effects rooting,callus,N/P, sucrose,6-BA and NAA concentrations on flowering in vitro Dianthus caryophyllus L.. The results of our research show that the plant takes root to repress flower bud inducement obviously,it is not very impact of floral bud induction whether differentiate callus or not.The results showed that the floral bud induction rate was obviously promoted when we adjusted the N of ammonium nitrate and P content,one of N/P ratio is 1N and 5P(phosphor concentration)can make highest rate of floral bud induction.N and P ratio of contention influence the floral color depth,and the normal growth of plant and leaf.When 6-BA and NAA are combined in level of 0.5mg.L~(-1)and 0.1 mg.L~(-1),the induction rate of flowering is highest at 72.3%.The optimal way for inducing floral buds is to preculture the plantets on new MS medium added 35g.L~(-1) sucrose,7g.L~(-1)agar,0.5mg.L~(-1)6-BA and 0.1mg.L~(-1)NAA.Plantlets,which were new stem growth from subcultured plantlets,were induced to from floral buds in 60 days. The rate of flower induction was about seventy percent.
The optimal medium for inducing floral buds is new MS medium added 35g.L~(-1) sucrose,7g.L~(-1)agar,0.5mg.L~(-1)6-BA and 0.1m NAA.The inefficient medium for inducing floral buds is MS medium added 30g.L~(-1)sucrose,6.5g.L~(-1)agar.This paper still study on the variation and correlations of physiological and biochemical of Dianthus caryophyllus L.during the flower bud induction.The results showed that the contention of soluble proteins,soluble sugar and incretion were higher in control group which floral bud induction is high than in treatment group which floral bud induction is low.
本文研究了不同浓度的6-BA、NAA,不同的琼脂浓度以及不同的封口材料对香石竹叶片和带有腋芽的茎段诱导的影响。研究发现,6-BA和NAA对香石竹叶片和腋芽的诱导率具有显著性差异。6-BA和NAA不同浓度的配比组合对叶片和腋芽诱导的影响也有很大程度的差别,当6-BA和NAA浓度分别为2.0mg.L~(-1)和0.5mg.L~(-1)的组合时,芽的诱导率最高达到11.2%和87.9%。培养基中不同的琼脂浓度对芽的诱导率形成显著性影响,当琼脂浓度为7g.L~(-1)时,芽的长势最好且发育出许多叶子。不同的封口材料对香石竹叶片和腋芽诱导效果显著,使用透气封口膜芽生长速度快,利于植株枝条和叶片的旺盛生长和形成。通过筛选,香石竹的腋芽在不同条件下的发芽率均要比叶片的发芽率高。
本文重点研究了生根、愈伤组织、N/P含量、蔗糖浓度、6-BA,NAA等对香石竹组培苗花芽发生和花发育的影响。研究发现,根对花芽形成有显著影响,生根明显抑制花芽诱导。是否分化愈伤组织对香石竹花芽诱导的影响不大。调整了MS培养基中的氮和磷含量,明显提高了花芽的诱导率,其中1N和5P的配比下花芽诱导率最高。在一定范围内培养基中蔗糖含量与花芽诱导率成正相关,但是过高的含量会产生玻璃化苗。6-BA(0.5mg.L~(-1))和NAA(0.1mg.L~(-1))配比时,花芽诱导率最高,达到72.3%。通过筛选,诱导香石竹花芽形成效果最好的培养方法是:在改良的MS基本培养基(1倍氮,5倍磷),蔗糖浓度提高到35g.L~(-1),并加入适宜浓度的6-BA(0.5mg.L~(-1))和NAA(0.1mg.L~(-1))可诱导香石竹试管苗在60天内开花。2个月内统计花芽诱导率为70%左右。
以诱导花芽形成效果最佳的最佳培养基作为处理组,同时设一对照组:MS,附加蔗糖30 g.L~(-1),琼脂浓度6.5g.L~(-1),pH5.8。研究香石竹花芽诱导过程中植株内部生理生化的变化和相关性。花芽诱导率高的处理,其可溶性蛋白质、可溶性糖、内源激素含量在整体上都明显高于花芽诱导率低的处理。
In vitro,in vitro flowering and internal variation of physiological and biochemical during flower bud differentiation for Dianthus caryophyllus L.through tissue culture, can build up Dianthus caryophyllus L.system of intermediate propagation,decrease the juvenile phases from a vegetative growth to a reproductive growth stage,inquiry into flower bud differentiation regulation is significance not only in theory but also in application.
The objective of this research is to study the effects of differ culture medium, 6-BA,NAA,differ Agar and closure film during the induction of lateral bud of Dianthus caryophyllus L..The results of our research show that the contrast of differ 6-BA and NAA concentration is very remarkable on the induction rate of lateral bud and leaf,also the effects of differ 6-BA and NAA concentration go together with ratio combination are very difference.When 6-BA and NAA are combined in level of 2.0 mg.L~(-1)and 0.5 mg.L~(-1),the induction rate of lateral bud is highest at 87.9%and 11.2%. The effect of differ Agar of culture medium is very remarkable on the induction of lateral bud and leaf of Dianthus caryophyllus L.As its Agar is 7g.L~(-1),the induction rate is highest,bud grow best and grow many leafs.The effect of differ closure film of culture medium is very remarkable on the induction of lateral bud and leaf of Dianthus caryophyllus L.Through using closure film,the effect on the induction of lateral bud of Dianthus caryophyllus L.also is significance,results shows that closure film can make bud grow rapidly and become a plant.
The mostly objective of this research is to study the effects rooting,callus,N/P, sucrose,6-BA and NAA concentrations on flowering in vitro Dianthus caryophyllus L.. The results of our research show that the plant takes root to repress flower bud inducement obviously,it is not very impact of floral bud induction whether differentiate callus or not.The results showed that the floral bud induction rate was obviously promoted when we adjusted the N of ammonium nitrate and P content,one of N/P ratio is 1N and 5P(phosphor concentration)can make highest rate of floral bud induction.N and P ratio of contention influence the floral color depth,and the normal growth of plant and leaf.When 6-BA and NAA are combined in level of 0.5mg.L~(-1)and 0.1 mg.L~(-1),the induction rate of flowering is highest at 72.3%.The optimal way for inducing floral buds is to preculture the plantets on new MS medium added 35g.L~(-1) sucrose,7g.L~(-1)agar,0.5mg.L~(-1)6-BA and 0.1mg.L~(-1)NAA.Plantlets,which were new stem growth from subcultured plantlets,were induced to from floral buds in 60 days. The rate of flower induction was about seventy percent.
The optimal medium for inducing floral buds is new MS medium added 35g.L~(-1) sucrose,7g.L~(-1)agar,0.5mg.L~(-1)6-BA and 0.1m NAA.The inefficient medium for inducing floral buds is MS medium added 30g.L~(-1)sucrose,6.5g.L~(-1)agar.This paper still study on the variation and correlations of physiological and biochemical of Dianthus caryophyllus L.during the flower bud induction.The results showed that the contention of soluble proteins,soluble sugar and incretion were higher in control group which floral bud induction is high than in treatment group which floral bud induction is low.
引文
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