银叶杜鹃组织培养技术体系研究
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摘要
本研究为探索银叶杜鹃(Rhododendron argyrophyllum)组织培养快速繁殖的有效途径，以银叶杜鹃3年生苗茎尖、无菌苗叶片、茎尖、茎段、根段为外植体，着重研究其培养的最佳培养基配方及培养程序，以及培养过程中防止污染和减轻褐变的措施，建立银叶杜鹃快速繁殖技术体系。
结果表明：
1)银叶杜鹃组织培养选取3月下旬到4月上旬刚长出的未展叶的幼嫩芽，切取2mm左右长的茎尖，为最理想的外植体。
2)在启动培养初期，即接种后的3天内每天在三角瓶内将茎尖转移位置，前4周每周转接一次，以后每4周转接一次，可有效减轻外植体的褐变。
3)启动培养：启动培养基为，Anderson(培养基)+6-BA0.1mg／L+NAA0.01mg／L，暗培养1周后转入光照培养，4周时启动率可达60.7％，AC(活性炭)对外植体褐变有一定的防止作用，加入0.1g／L的活性炭对启动培养效果较好。
4)丛生芽增殖：最适培养基为Anderson+6-BA1.0mg／L，丛生芽增殖最快，90天时增殖系数可达4.2。
5)适合试管苗伸长的培养基配方最佳组合为：Anderson +6-BA2.0mg／+NAA0.01 mg／L+AC1.0g／L。
6)适合银叶杜鹃生根的培养基配方组合为：Anderson+AC1.0g／L。
7)适合银叶杜鹃叶片愈伤组织诱导的培养基配方组合为：Anderson+6-BA1.0mg／L+2,4-D0.1 mg／L，暗培养2周后转入光照下培养。
8)银叶杜鹃无菌苗根段愈伤组织诱导的最佳组合为：Anderson+6-BA2.0mg／L+2,4-D0.1mg／L，暗培养3周后转入光照下培养。
9)银叶杜鹃无菌苗茎段丛生芽诱导的培养基配方组合为：Anderson+6-BA2.0mg／L，暗培养1周后转入光照下培养。
10)炼苗移栽基质以林下腐质土效果最好，移栽存活率可达85％。
The shoottips and stems of Rhododendron argyrophyllum were used as explants to establish, a rapid technique system of micropropagation. Root and leaf of Rhododendron argyrophyllum were used to find out the best medium compoisition of callus induction, hi the experiments , effective measures were found out to prevent the browning and contamination of shoot explants .The results were as follows:
1) Suitableculture conditions were:Day and night 25℃±1℃,14 hours illumination every day, and about 2000 lux illumination intensity and medium pH 5.3~5.6.
2) The best technique for initiation culture was Anderson+6-BA0.1mg/L + NAA0.01mg/L+,for a week of dark treatment. After 4 weeks culture, the rate of initiation could reach 60.7%.
3) For multiplication of shoot clumps, Anderson+6-BA1.0 mg/L was the best medium. After 12 weeks culture, the multiplication rate of shoot clumps could reach 4.2.
4) For elongation of the shoots, Anderson +6-BA2.0 mg/+NAA0.01 mg/L+AC1.0/Lwas the best medium. After 90 days culture, almost all the shoots' the length could reach 2cm.
5) For the rooting of shoots, Anderson +AClg/L was the best medium.
6) The best medium composition for callus induction from sterile seedling's leaf explants was Anderson +6-BA1.0 mg/L+2,4-D0.1 mg/L,for 2 weeks of dark treatment.
7) The best medium composition for callus induction from sterile seedling's root explant was Anderson+6-BA2.0mg/L+2,4- D0.1mg/L, for 3 weeks of dark treatment.
8)The best medium composition for shoots induction from sterile seeding's stalk was Anderson +6-BA2.0mg/L, for a week of dark treatment.
9) Activated charcoal was effective for preventing the explants from browning to a certain extent, 0.5g/L was the best initiation concentration.
10) For transplanting of plantlets, the soil full of rotten leaves in the groves was the best material.the survival rate of plantlets being 85%.
结果表明：
1)银叶杜鹃组织培养选取3月下旬到4月上旬刚长出的未展叶的幼嫩芽，切取2mm左右长的茎尖，为最理想的外植体。
2)在启动培养初期，即接种后的3天内每天在三角瓶内将茎尖转移位置，前4周每周转接一次，以后每4周转接一次，可有效减轻外植体的褐变。
3)启动培养：启动培养基为，Anderson(培养基)+6-BA0.1mg／L+NAA0.01mg／L，暗培养1周后转入光照培养，4周时启动率可达60.7％，AC(活性炭)对外植体褐变有一定的防止作用，加入0.1g／L的活性炭对启动培养效果较好。
4)丛生芽增殖：最适培养基为Anderson+6-BA1.0mg／L，丛生芽增殖最快，90天时增殖系数可达4.2。
5)适合试管苗伸长的培养基配方最佳组合为：Anderson +6-BA2.0mg／+NAA0.01 mg／L+AC1.0g／L。
6)适合银叶杜鹃生根的培养基配方组合为：Anderson+AC1.0g／L。
7)适合银叶杜鹃叶片愈伤组织诱导的培养基配方组合为：Anderson+6-BA1.0mg／L+2,4-D0.1 mg／L，暗培养2周后转入光照下培养。
8)银叶杜鹃无菌苗根段愈伤组织诱导的最佳组合为：Anderson+6-BA2.0mg／L+2,4-D0.1mg／L，暗培养3周后转入光照下培养。
9)银叶杜鹃无菌苗茎段丛生芽诱导的培养基配方组合为：Anderson+6-BA2.0mg／L，暗培养1周后转入光照下培养。
10)炼苗移栽基质以林下腐质土效果最好，移栽存活率可达85％。
The shoottips and stems of Rhododendron argyrophyllum were used as explants to establish, a rapid technique system of micropropagation. Root and leaf of Rhododendron argyrophyllum were used to find out the best medium compoisition of callus induction, hi the experiments , effective measures were found out to prevent the browning and contamination of shoot explants .The results were as follows:
1) Suitableculture conditions were:Day and night 25℃±1℃,14 hours illumination every day, and about 2000 lux illumination intensity and medium pH 5.3~5.6.
2) The best technique for initiation culture was Anderson+6-BA0.1mg/L + NAA0.01mg/L+,for a week of dark treatment. After 4 weeks culture, the rate of initiation could reach 60.7%.
3) For multiplication of shoot clumps, Anderson+6-BA1.0 mg/L was the best medium. After 12 weeks culture, the multiplication rate of shoot clumps could reach 4.2.
4) For elongation of the shoots, Anderson +6-BA2.0 mg/+NAA0.01 mg/L+AC1.0/Lwas the best medium. After 90 days culture, almost all the shoots' the length could reach 2cm.
5) For the rooting of shoots, Anderson +AClg/L was the best medium.
6) The best medium composition for callus induction from sterile seedling's leaf explants was Anderson +6-BA1.0 mg/L+2,4-D0.1 mg/L,for 2 weeks of dark treatment.
7) The best medium composition for callus induction from sterile seedling's root explant was Anderson+6-BA2.0mg/L+2,4- D0.1mg/L, for 3 weeks of dark treatment.
8)The best medium composition for shoots induction from sterile seeding's stalk was Anderson +6-BA2.0mg/L, for a week of dark treatment.
9) Activated charcoal was effective for preventing the explants from browning to a certain extent, 0.5g/L was the best initiation concentration.
10) For transplanting of plantlets, the soil full of rotten leaves in the groves was the best material.the survival rate of plantlets being 85%.
引文
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