High-performance Regeneration of Pyrethrum
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摘要:
目的 筛选出适宜白花除虫菊外植体的消毒方法、外植体诱导、继代培养及生根培养阶段适宜的培养基配方，建立白花除虫菊高效再生体系。 方法 以白花除虫菊尚未张开的花蕾作为再生体系的外植体材料，花蕾经过灭菌消毒以后，采用MS固体培养基，比较不同种类植物生长调节剂浓度配比对白花除虫菊花蕾诱导出芽、增殖及生根等关键环节的影响。 结果 白花除虫菊花蕾外植体最适宜的消毒条件为75%酒精处理30 s后用0.10%氯化汞溶液消毒10 min，之后用15%次氯酸溶液处理15 min； 白花除虫菊花蕾诱导芽的最适培养基为MS+2.0 mg·L−16-BA+0.5 mg·L−1TDZ + 0.2 mg·L−1IBA；白花除虫菊芽增殖的最适培养基为MS+ 1.0 mg·L−16-BA+ 0.1 mg·L−1TDZ+0.1 mg·L−1IBA；白花除虫菊生根的最适培养基为MS+ 0.1 mg·L−1IAA+ 0.1 mg·L−1IBA；炼苗移栽的最适基质为泥炭 ∶ 珍珠岩=6 ∶ 1，成活率达90%以上，移栽效果良好。 结论 基本建立了白花除虫菊高效再生体系，可为繁育白花除虫菊种苗提供技术支撑。 Abstract:Objective Appropriate methods for efficient tissue disinfection, explant induction, and culture medium formulation were established for a high-performance program to propagate pyrethrum. Method Unopened flower buds of white flower pyrethrum were sterilized and used as explants on an MS solid medium for the experiment. Effects of various plant growth regulators on bud induction, proliferation, and rooting of the seedlings were monitored. Result The optimum conditions for disinfecting the explants were found to be a treatment of 75% alcohol for 30 s followed by one of 0.10% mercuric chloride solution for 10 min and another of 15% hypochlorite for 15 min. For bud induction, the choice medium was formulated with MS + 2.0 mg·L−1 6-BA + 0.5 mg·L−1 TDZ+ 0.2 mg·L−1 IBA; for bud proliferation, MS + 1.0 mg·L−1 6-BA + 0.1 mg·L−1TDZ + 0.1 mg·L−1 IBA; for rooting, MS + 0.1 mg·L−1 IAA + 0.1 mg·L−1 IBA; and for transplanting seedlings, peat:perlite at 6:1. A survival rate greater than 90% as well as adequate transplanting was achieved. Conclusion The newly developed in vitro regeneration system materially lessened the pressure of the recently encountered white flower pyrethrum germplasm degradation. In addition, a supply of high-quality seedlings could be assured with the proposed propagation program. -
Key words:
- White flower pyrethrum /
- adventitious bud /
- regeneration system
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表 1 除虫菊小花蕾消毒试验结果
Table 1. Disinfection of pyrethrum small buds
处理
Treatment0.1%氯化汞处理
0.1% mercuric chloride treatment/min15%次氯酸钠溶液处理
15% sodium hypochlorite treatment/min污染率
Pollution rate/%褐化率
Brownning rate/%诱导率
Induction rate/%A1 10 10 50.00±2.72 a 16.67±2.72 d 22.22±4.16 d A2 10 15 31.11±4.16 c 8.89±1.57 e 64.44±1.57 a A3 10 20 47.78±1.57 b 36.67±2.72 a 32.22±1.57 c A4 15 10 46.67±2.72 b 30.00±2.72 c 38.89±4.16 c A5 15 15 28.89±1.57 d 27.78±1.57 c 50.00±2.72 b A6 15 20 24.44±1.57 d 33.33±2.72 b 58.89±4.16 a 同列数据后不同小写字母表示差异达0.05显著水平。表2～5同。
Data with different lowercase letters on the same column indicate significant difference at 0.05 level. Same for Table 2-5.
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表 2 除虫菊花蕾诱导培养基筛选
Table 2. Medium selection for pyrethrum bud induction
处理
Treatment植物生长调节剂
Plant growth regulator/ (mg·L−1)诱导率
Induction rate/%生长状态
Growth state6-BA TDZ IBA B1 1.0 0.1 0.2 65.56±1.57 d 诱导出不定芽，生长一般 B2 1.0 0.3 0.2 71.11±1.57 d 诱导出不定芽，较好 B3 1.0 0.5 0.2 80.00±2.72 b 诱导出不定芽，一般 B4 2.0 0.1 0.2 76.67±2.72 c 诱导出不定芽，一般 B5 2.0 0.3 0.2 87.78±4.16 a 诱导出不定芽，较好 B6 2.0 0.5 0.2 93.33±2.72 a 诱导出不定芽，生长健壮
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表 3 除虫菊不定芽增殖培养基筛选
Table 3. Medium selection for pyrethrum adventitious bud proliferation
处理
Treatment植物生长调节剂
Plant growth regulator/ (mg·L−1)增殖倍数
Proliferation rate生长状态
Growth state6-BA TDZ IBA C1 1 0.1 0.1 3.08±0.02 a 丛生芽生长健壮 C2 1 0.3 0.1 2.70±0.03 b 丛生芽生长较好 C3 1.5 0.1 0.1 2.50±0.03 d 丛生芽生长较好 C4 1.5 0.3 0.1 2.44±0.04 d 丛生芽生长一般 C5 2 0.1 0.1 2.54±0.04 c 丛生芽生长一般 C6 2 0.3 0.1 2.26±0.07 e 丛生芽生长较好，但部分有玻璃化
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表 4 除虫菊花蕾生根培养基的筛选结果
Table 4. Medium selection for pyrethrum seedling rooting
处理
Treatment植物生长调节剂
Plant growth regulator/ (mg·L−1)生根率
Rate of rooting/%生根情况
Rooting stateIAA IBA D1 0.1 0.1 98.89±1.57 a 根系生长状况良好 D2 0.1 0.2 86.67±2.72 b 根系生长状况好 D3 0.2 0.1 82.22±4.16 b 根系生长状况好 D4 0.2 0.2 80.00±2.72 b 根系生长状况一般
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表 5 除虫菊不同体积配比基质筛选结果
Table 5. Selected culture medium for pyrethrum propagation
处理
Treatment泥炭与珍珠岩体积配比
Proportions of peat soil and perlite成活率
Rate of survival/%株高
Plant height/cm叶数
Leaf numberE1 1∶1 75.56±6.85 b 5.77±0.29 b 5.67±0.47 b E2 2∶1 73.33±2.72 b 5.83±0.12 b 6.33±1.25 b E3 4∶1 85.56±6.85 b 6.20±0.16 b 7.00±1.41 b E4 6∶1 96.67±2.72 a 6.43±0.21 a 7.33±1.25 a
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