茉莉离体快繁体系的建立
摘要
茉莉[Jasminum sambac (L.) Aiton]是木樨科茉莉属常绿小灌木,原产波斯湾一带,是我国香料工业的主要原料之一,主要用来窨制茶叶、提取精油等。其花色洁白,香气宜人,近年来被广泛应用于家庭摆设和庭园观赏,深受国人喜欢。茉莉作为一种具有极高的观赏和经济价值的花卉,其生产、开发和综合利用前景广阔。本文以茉莉萌动腋芽为外植体,研究了基本培养基、生长素、细胞分裂素等因素对其试管苗启动、增殖、生根等的影响,初步建立了茉莉的离体快繁体系。主要结果如下：
启动培养：研究了不同灭菌时间、培养基种类(MS、M1、WPM)、不同细胞分裂素(TDZ、6-BA、KT)与生长素(NAA、IBA)组合以及不同光温条件对茉莉腋芽启动培养的影响。结果表明：0.1%升汞处理8 min灭菌效果最好。WPM+6-BA2.0mg/L+IBA 0.1 mg/L是最佳的启动培养基,诱导率为96.75%、平均芽数为1.97、平均芽长为3.80 cm、增殖系数为2.79。最适合培养条件为温度35℃,光照强度6000 lx。试验也发现枝条基部外植体诱导获得的试管苗生长状况最好,5-9月份均适宜取材。
增殖培养：研究了不同基本培养基、碳源、温度及不同生长素、细胞分裂素、GA3浓度对茉莉试管苗增殖培养的影响。结果表明：WPM+3%蔗糖为最佳基本培养基。35℃下6-BA 1.0 mg/L+IBA 0.2 mg/L组合对试管苗的增殖效果最好,增殖系数为5.31,平均芽数为1.32、平均芽长为4.87 cm,生长量为6.43 cm。GA3和CH对试管苗的增殖培养有抑制作用。0.2 mmol/L铁盐和镁盐可以提高试管苗叶绿素含量,降低试管苗黄化现象。
生根培养：以茉莉继代试管苗为试材,研究了不同培养基(MS、1/2MS、WPM和1/2WPM)、不同生长素(IBA和NAA)、生根方式(一步生根和两步生根)、不同继代次数(1-6次继代培养)以及培养基支持物(琼脂、珍珠岩、石英砂和蛭石)对茉莉生根的影响。结果表明：最佳生根培养基为1/2WPM,最佳生根培养方式为1/2WPM+NAA 1.0 mg/L处理7d后转入空白培养基进行两步法生根,生根率为98.41%；经1-4次继代的试管苗生根效果好于5或6次继代的试管苗；以蛭石为培养基支持物诱导生根效果最好,生根率达到100%,移栽成活率97.14%,根系活力、POD和SOD活性也均高于其他培养基支持物。
Jasmine [Jasminum sambac (L.)Aiton], a evergreen undershrub belonging to Oleaceae family, originated India. Jasmine, one kind of main perfume material, was used at scenting tea, extracting essential oil. Because of the white flower and pleasant fragrance, Jasmine was called the most fragrant flower in world, so it has been widely applied for ornamental at family and courtyard recent years and was deeply liked by people. It is obvious that Jasmine has much more ornamental and economic value. Jasmine has broad prospects on production, development and application. In this article, we discussed the effects basic mediums, auxins, CTks, and temperature on the proliferation in vitro of Jasmine.
1. Adventitious buds inducted culture
The effects of different sterilization time, basic medium (MS、M1、WPM), different concentrations of cytokinins(TDZ,6-BA、KT) and auxins (NAA、IB A) on induction culture of Jasmine were studied. Meanwhile, effects of light intensity and temperature on induction of Jasmine were investigated. The best effect of sterilization was 70% alcohol for 30 seconds,0.1% HgCl2 10 minutes for explant. Results showed that the highest bud induction rate was obtained at the WPM medium supplemented with 2.0 mg/L 6-BA and 0.1 mg/L IBA. Induction rate reached 96.75%, the average number of shoot was 1.97, the shoot was as long as 3.8 cm on average, and proliferation coefficient was up to 2.79. The shoot grew well under light intensity of 6000 lx at 35℃. Shoot cultured from the basal explant had a best stage. Explant grew on May to September was availably for experiment.
2. Proliferation culture
The effects of different basic mediums, carbon sources, on proliferation of Jasmine at different temperature were investigated. Moreover, effects of different concentrations of NAA/IBA and 6-BA/KT, GA3, CH on proliferation were investigated. Results showed that the best basic medium was the WPM medium supplemented with 3% sucrose. The best proliferation with a shoot proliferation coefficient of 5.31 was observed at the WPM medium supplemented with 1.0 mg/L 6-BA and 0.2 mg/L IBA at 35℃. Under the same condition, the average number of shoot was 1.32 and the average length of shoot reached 4.87 cm and the growth was as high as 6.43. However, GA3 and CH played negative role on proliferation. Supplemented with 0.2 mmol/L ferric salt and magnesium salt, Chlorophyll content of shoot was increased. Meanwhile, etiolation symptom of leaf was reduced.
3. Rooting culture
The effects of different medium (MS、1/2MS、WPM and 1/2WPM),auxin (IBA and NAA), rooting culture mode (one-step rooting and two-step rooting), plantlets subcultured in vitro for one to six times and supporting materials (agar, perlite, quartz sand and vermiculite) on Jasminum rooting were studied. The results showed that 1/2WPM optimized rooting for Jasmine. Two-step rooting was the optimal rooting culture mode, with 98.41% of rooting rate. Plantlet was pre-cultured on the rooting medium of 1/2WPM+NAA 1.0 mg/L for 7 to 10 days, then transferring pre-cultured shoots to 1/2WPM without supplement of any plant growth regulators. Rooting ability of the shoots subcultured in vitro 1 to 4 times was well. Besides, the effects of agar, perlite, quartz sand and vermiculite as supporting materials were studied. Vermiculite was the best supporting material; rooting rate was up to 100%; the survive rate of transplant seedlings reached 97.14%; root vigor, POD activity and SOD activity were higher than the other mediums, respectively.
启动培养：研究了不同灭菌时间、培养基种类(MS、M1、WPM)、不同细胞分裂素(TDZ、6-BA、KT)与生长素(NAA、IBA)组合以及不同光温条件对茉莉腋芽启动培养的影响。结果表明：0.1%升汞处理8 min灭菌效果最好。WPM+6-BA2.0mg/L+IBA 0.1 mg/L是最佳的启动培养基,诱导率为96.75%、平均芽数为1.97、平均芽长为3.80 cm、增殖系数为2.79。最适合培养条件为温度35℃,光照强度6000 lx。试验也发现枝条基部外植体诱导获得的试管苗生长状况最好,5-9月份均适宜取材。
增殖培养：研究了不同基本培养基、碳源、温度及不同生长素、细胞分裂素、GA3浓度对茉莉试管苗增殖培养的影响。结果表明：WPM+3%蔗糖为最佳基本培养基。35℃下6-BA 1.0 mg/L+IBA 0.2 mg/L组合对试管苗的增殖效果最好,增殖系数为5.31,平均芽数为1.32、平均芽长为4.87 cm,生长量为6.43 cm。GA3和CH对试管苗的增殖培养有抑制作用。0.2 mmol/L铁盐和镁盐可以提高试管苗叶绿素含量,降低试管苗黄化现象。
生根培养：以茉莉继代试管苗为试材,研究了不同培养基(MS、1/2MS、WPM和1/2WPM)、不同生长素(IBA和NAA)、生根方式(一步生根和两步生根)、不同继代次数(1-6次继代培养)以及培养基支持物(琼脂、珍珠岩、石英砂和蛭石)对茉莉生根的影响。结果表明：最佳生根培养基为1/2WPM,最佳生根培养方式为1/2WPM+NAA 1.0 mg/L处理7d后转入空白培养基进行两步法生根,生根率为98.41%；经1-4次继代的试管苗生根效果好于5或6次继代的试管苗；以蛭石为培养基支持物诱导生根效果最好,生根率达到100%,移栽成活率97.14%,根系活力、POD和SOD活性也均高于其他培养基支持物。
Jasmine [Jasminum sambac (L.)Aiton], a evergreen undershrub belonging to Oleaceae family, originated India. Jasmine, one kind of main perfume material, was used at scenting tea, extracting essential oil. Because of the white flower and pleasant fragrance, Jasmine was called the most fragrant flower in world, so it has been widely applied for ornamental at family and courtyard recent years and was deeply liked by people. It is obvious that Jasmine has much more ornamental and economic value. Jasmine has broad prospects on production, development and application. In this article, we discussed the effects basic mediums, auxins, CTks, and temperature on the proliferation in vitro of Jasmine.
1. Adventitious buds inducted culture
The effects of different sterilization time, basic medium (MS、M1、WPM), different concentrations of cytokinins(TDZ,6-BA、KT) and auxins (NAA、IB A) on induction culture of Jasmine were studied. Meanwhile, effects of light intensity and temperature on induction of Jasmine were investigated. The best effect of sterilization was 70% alcohol for 30 seconds,0.1% HgCl2 10 minutes for explant. Results showed that the highest bud induction rate was obtained at the WPM medium supplemented with 2.0 mg/L 6-BA and 0.1 mg/L IBA. Induction rate reached 96.75%, the average number of shoot was 1.97, the shoot was as long as 3.8 cm on average, and proliferation coefficient was up to 2.79. The shoot grew well under light intensity of 6000 lx at 35℃. Shoot cultured from the basal explant had a best stage. Explant grew on May to September was availably for experiment.
2. Proliferation culture
The effects of different basic mediums, carbon sources, on proliferation of Jasmine at different temperature were investigated. Moreover, effects of different concentrations of NAA/IBA and 6-BA/KT, GA3, CH on proliferation were investigated. Results showed that the best basic medium was the WPM medium supplemented with 3% sucrose. The best proliferation with a shoot proliferation coefficient of 5.31 was observed at the WPM medium supplemented with 1.0 mg/L 6-BA and 0.2 mg/L IBA at 35℃. Under the same condition, the average number of shoot was 1.32 and the average length of shoot reached 4.87 cm and the growth was as high as 6.43. However, GA3 and CH played negative role on proliferation. Supplemented with 0.2 mmol/L ferric salt and magnesium salt, Chlorophyll content of shoot was increased. Meanwhile, etiolation symptom of leaf was reduced.
3. Rooting culture
The effects of different medium (MS、1/2MS、WPM and 1/2WPM),auxin (IBA and NAA), rooting culture mode (one-step rooting and two-step rooting), plantlets subcultured in vitro for one to six times and supporting materials (agar, perlite, quartz sand and vermiculite) on Jasminum rooting were studied. The results showed that 1/2WPM optimized rooting for Jasmine. Two-step rooting was the optimal rooting culture mode, with 98.41% of rooting rate. Plantlet was pre-cultured on the rooting medium of 1/2WPM+NAA 1.0 mg/L for 7 to 10 days, then transferring pre-cultured shoots to 1/2WPM without supplement of any plant growth regulators. Rooting ability of the shoots subcultured in vitro 1 to 4 times was well. Besides, the effects of agar, perlite, quartz sand and vermiculite as supporting materials were studied. Vermiculite was the best supporting material; rooting rate was up to 100%; the survive rate of transplant seedlings reached 97.14%; root vigor, POD activity and SOD activity were higher than the other mediums, respectively.
引文
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