摘要组织培养是获得植物再生苗的常用手段之一，具有高效且经济的优势。为了适应需求，建立良好的常山胡柚植株再生体系是当务之急。此项研究以常山胡柚（Citrus changshan-huyou Y. B. Chang, sp. Nov）的幼茎、幼叶和幼胚为外植体，进行组织培养，观察并记录了在不同植物激素条件下，离体培养常山胡柚幼茎、幼叶、幼芽存活率、愈伤化的影响及幼胚的生长状况。

实验结果表明：不同浓度植物激素诱导培养胡柚幼叶，在15天左右，少数叶片出现黄化死亡，部分叶片出现卷缩，进一步形成愈伤组织；不同浓度植物激素诱导培养幼茎，20~30天后，部分幼茎黄化死亡，部分幼茎出现膨大情况，但未形成明显的愈伤组织，极小部分在幼茎一端直接形成芽。幼胚培养过程中，萌发过程较为缓慢，且幼胚萌发率不足50%。在较低浓度6-BA（0.25~0.75 mg/L）条件下幼胚可以直接分化出丛生芽；而在较高浓度6-BA条件下，幼胚仅脱分化为愈伤组织。而且，这些愈伤组织转入含有较低浓度6-BA的培养基MS+0.5 mg/L 6-BA上，也能再生出芽。将幼胚形成的丛生芽，转接到MS+0.1~0.2 mg/mL IAA，或者MS+0.1~0.2 mg/mL IAA+0.5 mg/mL 6-BA的培养基上，丛生芽均能生根，在培养20~30天后，根长可以达到2~3 cm，最后形成完整植株。这是常山胡柚快速繁殖的第一项成功试验。71042

Abstract Tissue culturing is an efficient and economical method of plant regeneration. In order to meet requirements, establishing a regeneration system of Citrus Changshan-Huyou is very necessary now. In this study, the young stems, young leaves and immature embryos were chosen as explants. The different concentrations of plant hormone condition in the culture medium were investigated in tissue culture of Citrus Changshan-Huyou.

The results were shown as follows. During young leaves of Citrus cultured on medium with different concentrations of plant hormone, some leaves turned yellow and dead after 15 days or so. Besides, some leaves curled, and turned to callus after culture. Some young stems turned yellow and dead after 20~30 days or so, others swelled on both ends and all did not form a visible callus during culture period. However, several young stem had buds on cut ends. During the immature embryos’ culturing, the germination process is very slow and it showed a low germination rate less than 50%. Buds from the immature embryos were produced on medium with lower concentration of 6-BA (0.25~0.75 mg/L), while calli were induced on the medium with higher concentration 6-BA. When the calli were transferred into MS+0.5 mg/L 6-BA medium, they could also dedifferentiate into buds. Transferred the cluster buds onto medium of MS+0.1~0.2 mg/mL IAA or MS+0.1~0.2 mg/mL IAA+0.5 mg/mL 6-BA, the cluster buds can produce roots, and the roots lengths can reach 2~3 cm after 20~30 days and the complete plants were got. To our knowledge, this is the first report on plant regeneration of Citrus Changshan-Huyou via tissue culture.

毕业论文关键词：常山胡柚； 愈伤化； 丛生芽； 植物激素； 组织培养

Key words:  Citrus changshan-huyou;  Callus;  Buds;  Plant hormone;  Tissue culture

目录

目录 3

引言 4

1 材料与方法 4

1.1  实验材料 4

1.2  培养方法 5

1.1.1 外植体消毒 5

1.2.2 接种 5

1.2.3 幼胚离体培养 5

2  结果与分析 5

    2.1常山胡柚幼叶培养结果