【作者】 王惠珍；

【导师】 王小菁；

【作者基本信息】 华南师范大学 ， 植物学， 2002， 硕士

【摘要】 本文研究了穗冠花Celosia cristata（phumosa）愈伤组织诱导和影响愈伤组织生长及花色素苷积累的培养条件。比较了来自不同苗龄的不同外植体愈伤组织诱导率、愈伤组织生长量及花色素苷含量。筛选并建立了生长快速稳定的产花色素苷的愈伤组织体系JSA10和JSA25。并以JSA10为试验材料，就影响愈伤组织生长和花色素苷积累的条件进行了筛选。主要实验结果如下： 穗冠花种子播种于1／2MS培养基上萌发，无菌苗健壮，对诱导愈伤和花色素苷的积累都有利。以20d或30d龄的无菌苗下胚轴作外植体，效果好，愈伤组织生长量大，花色素苷的含量也高。MS+2，4-D 0.1 mg／L+BA 0.5 mg／L培养基适合愈伤组织诱导和继代培养。 经继代培养筛选分离出JSA10洋红和JSA25橙红两种愈伤组织系列。JSA10愈伤组织生长能力和积累花色素苷能力均比JSA25强。JSA10和JSA25的愈伤组织生长曲线均呈“S”形，JSA10在第8d进入对数生长期，8d～12d是愈伤组织的快速生长期，14d～18d是愈伤组织生长缓慢期，20d以后是愈伤组织衰老期。花色素苷的积累在愈伤组织生长缓慢期（14d～18d）达到高峰。JSA10愈伤组织生长好，花色素苷含量稳定，花色素苷产量高峰期相对较长。JSA25与JSA10相比，生长周期较短，进入对数生长期提前2d，增长高峰期短，较快进入生长缓慢期，花色素苷的积累高峰也是在生长缓慢期。 以JSA10继代培养和进行各种因素的影响实验，发现，2，4-D_0.2+BA_1～2；2，4-D_0.2+KT₅；NAA₂+BA₁为适合愈伤组织生长和花色素苷积累的几种组合。其中以2，4-D_0.2+BA₂为最好，愈伤l 组织鲜重增长最快，花色素苦的产量达到最高。 在试验的 10种基本培养基中，MS、BS、LS三种培养基适合 穗冠花愈伤组织生长和花色素昔的积累，其中LS最好。 在 7种所试验的碳源中，以蔗糖效果最好，30～40g／L的浓 度对愈伤组织生长和花色素苦的积累较为有利。浓度高至 90 g／L 时花色素苦的含量最高，但不利于愈伤组织的生长。 培养基中的总含氮量以 30 m mol几为宜，浓度大时促进花色 素芳的积累，但对愈伤组织的生长不利。以 NH／作唯一氮源时， 会对愈伤组织的生长起抑制作用。 光照对愈伤组织的生长和花色素苦的积累有明显的促进作 用。蓝光和红光都促进愈伤组织的生长；而对花色素昔的积累， 蓝光有效。更多还原

【Abstract】 Culture conditions for callus induction and anthocyanin accumulation of Celosia cristata ’phumosa’ were studied in this paper. The induction rate, callus growth and anthocyanin accumulation of different explants isolated from the seedings in different age were compared for the selecting of rapid-growing and steadily-anthocyanin-accumulating lines. Two callus lines JSA10 and JSA25 were established and the former was used to investigate the factors regulating callus growth and pigmentation. The main results are as follows:Seeds of Celosia cristata ’phumosa’ were germinated on 1/2 MS medium and the seedings with different age were harvested. The explants of for root, hypocotyl and cotyledon were cultured. Among them the hypocotyls from 20d or 30d seedings was the best for callus forming and anthocyanin production. The proper medium was MS supplementing with 0.1mg/L 2,4-D and 0.5mg/L BA in callus induction and subculture.JSA10 (magenta colored line) and JSA25 (orange colored line) were selected from subculture. Both of them showed similar "S-type" growth curve, of which JSA10 was better in callus growth and pigmentation, and the period of anthocyanin production was longer. The beginning of exponential growth phase for JSA10 was 8th day, with a rapid growth phase from 8th to 12th day followed by a slower growth phase from 14th to 18th day, then entered a senescence phase after 20th day. The anthocyanin accumulation reached its highest level when the growth was slower (14th to 18th day). In contrast with JSA10, JSA25 showed a shorter growth cycle and entered exponential growth phase 2 days earlier and had a short rapid growth phase. Its peak of anthocyanin accumulation was matched the slower growth phase.JSA10 was subcultured and the effect of plant growth regulators on callus growth and anthocyanin production was examined. 2,4-Do.2+BA1-2, 2,4-D0.2+KT5, NAA2+BA1 were suitable and the best combination was 2,4-Do.2+BA2. Effects of different basic media, carbon sucrose and total nigrogen concentration on callus growth and anthocyanin production of JSA10 were compared, of which, MS, B5 and LS were optimal for callus growth and anthocyanin production concentrations of 30-40g/L was fitness both for callus growth and for pigmentation. Sucrose of 90g/L in the medium could produce more anthocynin but inhibited callus growth. When the medium containing 30mmol/L nitrogen callus growth and pigmenting appeared better. Higher concentration of nitrogen increased more anthocyanin but inhibited callus growth. Growth suppression was observed with NH/ was only nitrogen source added in the medium.Stronger illumination was necessary in callus growth and anthocyanin production of Celosia cristata ’phumosa’, and effects of light quality were obvious, blue and red lights enhanced callus growth and only blue light showed significant effect on anthocyanin production.更多还原