蚬壳花椒组培快繁及其影响因子的研究
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摘要
蚬壳花椒(Zanthoxylum dissitum Hemsl)是一种药用植物,含有多种生物碱成份,是“妇科千金片”主要成分之一,具有十分重要的开发利用价值。
以蚬壳花椒当年生种子为材料,采用单因素、双因素和正交试验设计,研究了蚬壳花椒种子不同处理与灭菌时间对离体胚萌发率的影响,分析了不同基本培养基、不同种类激素及其配比、生物素浓度对蚬壳花椒组培苗的增殖及生长的影响,探讨了不同基本培养基、激素对组培苗生根的影响,进行了基质、容器等对移栽、扦插成活率影响的试验。研究结果表明：
(1)以蚬壳花椒种胚为外植体,70%酒精消毒15s,再用0.1%氯化汞进行表面消毒8min,最佳诱导培养基为MS+6-BA1mg/L+NAA0.4mg/L,诱导率可达86.67%。
(2)初代培养MS+ZT2.0mg/L+NAA0.2mg/L,培养条件为光强控制在3000-40001x,温度为25-30℃,pH5.8-6.0,蔗糖浓度确定为30g/L。
(3)继代培养中基本培养基为MS,激素为6-BA与NAA,使用6-BA的浓度不宜超过2.0mg/L,NAA的浓度范围在1.0mg/L,适宜继代培养基配方为MS+6-BA0.5mg/L+NAA 0.05mg/L+生物素1.0mg/L,增殖系数可达3.9。
(4)生根培养中,以1/3MS为基本培养基,生根培养的激素为IBA0.5mg/L+NAA0.05mg/L,培养温度29℃左右有利于组培苗根的诱导,生根率可达78%。
(5)移栽、扦插培养过程中,根据蚬壳花椒原始生境土壤条件分析的数据配置移栽、扦插基质。生根组培苗在1000倍液的杀菌剂中浸泡2min后,移栽到基质为黄心土+糠壳灰+细沙(1：1：1)的一次性杯子中,组培苗移栽成活率达80%。未生根组培苗经1000倍液的杀菌剂中浸泡2min后,IBA浸泡0.5h后,扦插到珍珠岩+泥炭土(3：1)基质中,扦插成活率49%。
通过系统研究确定蚬壳花椒组培快繁技术规程为：
材料处理→接种及诱导培养→建立无性繁殖体系→无菌苗增殖→生根壮苗培养→组培苗移栽、扦插。
Zanthoxylum dissitum Hemsl is a medical plant which contained various of alkaloid that had significant value in use.
In this thesis, we did experiments with the seeds of current year after certain treatment, adopted single-factor, double factors and orthogonal design methods, and studied the effects of the different treatments and sterilization time to the germinate ratio. We analyzed the effects of the proliferation、growth、rooting transplant and cutting caused by the different basal mediums、kinds and concentrations of hormones、substrates and so on. From all the experiments,the main results in this research were as following:
(1)Embryo of seeds was the optimal explants which was soaked in 70% alcohol 15s, then transferred into 0.1% HgCl2 8min to sterilize, Embryo was rarely polluted. MS+6-BA 1mg/L+NAA 0.4mg/L was suitable for embryo. The efficiency of callus induction reached 86.67%.
(2)The primary culture was MS+ZT2.0mg/L+NAA0.2mg/L, light was 3000～40001x, temperature was 25-30℃, pH was 5.8～6.0, concertration of sucrose was 30g/L.
(3)The medium and hormones of proliferation was MS+6-BA0.5mg/L+NAA 0.05mg/L+auxin1.0mg/L. The number of proliferation reached 3.9.
(4)Appropriate high temperature was benefited to rooting and 29℃was the optimal temperature for rooting. Roots ware induced on the media of 1/3MS +IBA0.5 mg/L+NAA0.05 mg/L.The percent of rooting reached 78%.
(5)The optimum transplant substance was soil+ plant ash+sand (1:1:1) and the ratio of survival reached 80%,The suitable cutting substance was pearlite+ soil (3:1)and the percent of survival reached 49%.
The technical rules of propagate Zanthoxylum dissitum Hemsl by tissue culture:
treating material→inoculating and induce culture→multiplying axenic plants→culturing strong plants with roots→transplanting and cutting.
以蚬壳花椒当年生种子为材料,采用单因素、双因素和正交试验设计,研究了蚬壳花椒种子不同处理与灭菌时间对离体胚萌发率的影响,分析了不同基本培养基、不同种类激素及其配比、生物素浓度对蚬壳花椒组培苗的增殖及生长的影响,探讨了不同基本培养基、激素对组培苗生根的影响,进行了基质、容器等对移栽、扦插成活率影响的试验。研究结果表明：
(1)以蚬壳花椒种胚为外植体,70%酒精消毒15s,再用0.1%氯化汞进行表面消毒8min,最佳诱导培养基为MS+6-BA1mg/L+NAA0.4mg/L,诱导率可达86.67%。
(2)初代培养MS+ZT2.0mg/L+NAA0.2mg/L,培养条件为光强控制在3000-40001x,温度为25-30℃,pH5.8-6.0,蔗糖浓度确定为30g/L。
(3)继代培养中基本培养基为MS,激素为6-BA与NAA,使用6-BA的浓度不宜超过2.0mg/L,NAA的浓度范围在1.0mg/L,适宜继代培养基配方为MS+6-BA0.5mg/L+NAA 0.05mg/L+生物素1.0mg/L,增殖系数可达3.9。
(4)生根培养中,以1/3MS为基本培养基,生根培养的激素为IBA0.5mg/L+NAA0.05mg/L,培养温度29℃左右有利于组培苗根的诱导,生根率可达78%。
(5)移栽、扦插培养过程中,根据蚬壳花椒原始生境土壤条件分析的数据配置移栽、扦插基质。生根组培苗在1000倍液的杀菌剂中浸泡2min后,移栽到基质为黄心土+糠壳灰+细沙(1：1：1)的一次性杯子中,组培苗移栽成活率达80%。未生根组培苗经1000倍液的杀菌剂中浸泡2min后,IBA浸泡0.5h后,扦插到珍珠岩+泥炭土(3：1)基质中,扦插成活率49%。
通过系统研究确定蚬壳花椒组培快繁技术规程为：
材料处理→接种及诱导培养→建立无性繁殖体系→无菌苗增殖→生根壮苗培养→组培苗移栽、扦插。
Zanthoxylum dissitum Hemsl is a medical plant which contained various of alkaloid that had significant value in use.
In this thesis, we did experiments with the seeds of current year after certain treatment, adopted single-factor, double factors and orthogonal design methods, and studied the effects of the different treatments and sterilization time to the germinate ratio. We analyzed the effects of the proliferation、growth、rooting transplant and cutting caused by the different basal mediums、kinds and concentrations of hormones、substrates and so on. From all the experiments,the main results in this research were as following:
(1)Embryo of seeds was the optimal explants which was soaked in 70% alcohol 15s, then transferred into 0.1% HgCl2 8min to sterilize, Embryo was rarely polluted. MS+6-BA 1mg/L+NAA 0.4mg/L was suitable for embryo. The efficiency of callus induction reached 86.67%.
(2)The primary culture was MS+ZT2.0mg/L+NAA0.2mg/L, light was 3000～40001x, temperature was 25-30℃, pH was 5.8～6.0, concertration of sucrose was 30g/L.
(3)The medium and hormones of proliferation was MS+6-BA0.5mg/L+NAA 0.05mg/L+auxin1.0mg/L. The number of proliferation reached 3.9.
(4)Appropriate high temperature was benefited to rooting and 29℃was the optimal temperature for rooting. Roots ware induced on the media of 1/3MS +IBA0.5 mg/L+NAA0.05 mg/L.The percent of rooting reached 78%.
(5)The optimum transplant substance was soil+ plant ash+sand (1:1:1) and the ratio of survival reached 80%,The suitable cutting substance was pearlite+ soil (3:1)and the percent of survival reached 49%.
The technical rules of propagate Zanthoxylum dissitum Hemsl by tissue culture:
treating material→inoculating and induce culture→multiplying axenic plants→culturing strong plants with roots→transplanting and cutting.
引文
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