【作者】 叶建伟；

【导师】 尹增芳；

【作者基本信息】 南京林业大学 ， 植物学， 2007， 硕士

【摘要】 本论文以宣木瓜的种子为材料,进行了组织培养和耐盐性两个方面的实验。1.组织培养方面的主要结论如下:1.1不定芽快速繁殖、诱导生根:与KT和TDZ相比,BA对不定芽增殖具有更好的效果。NAA与IBA两种生长调节物质对不定芽增殖的影响没有明显的差异。木瓜不定芽在BA0.5 mg/l +NAA0.2 mg/l培养基中,增殖系数最高。降低MS大量元素对不定芽增殖具有负作用。在1/2MS+IBA0.2组合中,不定芽的根诱导率较高、根较长。1.2愈伤组织的诱导和继代培养:1.2.1愈伤组织的诱导①在子叶诱导实验中,2,4-D能比NAA更高效的诱导出愈伤组织,愈伤生长量大,而且经过继代培养后仍能保持较好的长势。2,4-D与KT组合的效果好于2,4-D与BA的组合。筛选试验表明:MS+2,4-D1mg/l+KT0.1mg/l对子叶愈伤诱导及其生长效果较好。②胚轴愈伤组织诱导率达到87%,而子叶较低为69%。虽然NAA诱导的效果较好,但愈伤组织生长速度较慢;而2,4-D则能高效地诱导出愈伤,但所诱导的愈伤容易出现玻璃化现象。提高NAA,2,4-D和BA的浓度,愈伤呈现水浸状的比例和程度都会相应的提高。所设计的组合中,2,4-D 0.1mg/l+BA 0.3 mg/l对胚轴愈伤的诱导效果较好。③暗培养能提高愈伤组织的生长量.随着黑暗培养时间的延长,愈伤组织松散程度也不断上升,子叶颜色由浓绿色逐步向淡黄色过渡。1.2.2愈伤组织的继代培养①在固体培养基中,子叶愈伤组织继代培养可以根据目的不同选取不同的继代培养基。长期继代培养,将愈伤组织继代培养在NAA1.0mg/l+KT0.5mg/l的培养基或NAA0.5 mg/l与TDZ 2mg/l的组合中,可得到较好的效果。如需短期内获得较多的愈伤组织,可将愈伤组织在2,4-D1.0mg/l+KT0.1mg/l中培养。②子叶愈伤在液体培养条件下,生长量较低,效果较差。③胚轴愈伤在NAA1mg/l+KT 0.3mg/l培养基中的生长量最大。④维生素C对愈伤组织降低褐化的作用较小,但能促进愈伤组织变绿。1.3不定芽的诱导发生:子叶诱导不定芽的培养基中需要添加一定量的TDZ和NAA。不定芽有玻璃化现象,在TDZ 0.5 mg/l + NAA 1.0mg/l的培养基中得到的不定芽发育较好。2.不同NaCl浓度对种子和愈伤组织的生长的影响,主要结论如下:2.1愈伤组织:当NaCl浓度较低时,愈伤组织的生长量未受到较大影响、MDA含量较低,POD、SOD活性提高,细胞内可溶性糖、脯氨酸含量升高。而在高浓度NaCl的胁迫下,愈伤组织的生长具有明显的抑制作用、MDA含量升高,而POD、SOD活性开始降低、可溶性糖含量也降低,但是脯氨酸含量一直升高。2.2幼苗随着NaCl浓度的升高,种子的萌发率和幼苗根系活力呈递减趋势;除了在0.5‰NaCl处理的幼苗中,其叶绿素含量高于对照,其余处理均低于对照;幼苗的SOD、POD活性变化趋势较一致,先升高后降低,在1‰NaCl浓度具有最好的活性。MDA含量随NaCl处理浓度的升高而升高。更多还原

【Abstract】 Take the seeds of Chaenomeles speciosa Nakai as plant material; we carried out researches about tissue culture and salt tolerant.1. The most important conclusion about tissue culture, as follows:1.1 Multiply and root of adventitious:Compared KT and TDZ, BA has a better effect on multiply adventitious buds of Chaenomeles speciosa Nakai. The effect of NAA and IBA on multiply adventitious buds is similar. The explants cultured on medium supplemented with BA0.5 mg/l +NAA0.2 mg/l have the highest multiply ability. Reducing the content of MS macro element has a negative effect on multiply adventitious buds. The adventitious buds cultured on medium contained 1/2MS+IBA0.2 mg/l have highest induction root rate, and inducted roots is longer than other combination medium.1.2 Induction and subculture of callus tissue:1.2.1 Callus induction①In the experiment of cotyledons, 2,4-D has a better effect on inducing and subculture callus tissue than that of NAA. A satisfactory result of callus tissue induction and callus tissue growth was achieved on MS medium supplemented with 1mg/l 2,4-D and 0.1mg/l KT.②Callus tissue induction rate of hypocotyls was 87%, and it was higher than that of cotyledons which was 69%. The proportion and the degree of callus vitrifaction would scaled up as increasing the concentration of auxin and BA. Best callus of hypocotyls were obtained on MS medium contained 0.1mg/l 2,4-D and 0.3mg/l BA .③Dark culture incentives callus tissue growth. Longer the days of dark culture, the looser of callus tissue. The colors of cotyledons transferred from dark green to light yellow with increasing dark culture.1.2.2 Subculture of callus tissue①On solid medium, the subculture of cotyledons has different culture methods. If the subculture with long time, callus tissue was cultured on medium supplemented with NAA1.0mg/l+KT0.5mg/l or NAA0.5 mg/l + TDZ 2mg/l, which can obtain better result. On the other hand, callus tissue can growth quickly on medium contained 2,4-D 0.1 mg/l +BA 0.3mg/l and we can get more callus tissue in a short time.②Cultured in liquid medium, callus tissue of cotyledons grew slowly.③Callus tissue of hypocotyls growth most rapidly when cultured on medium contained NAA1mg/l+KT0.3mg/l.④The effect of Vc on reducing browning of callus tissue of Chaenomeles speciosa Nakai was less, but promoted colors of callus tissue turn to green.1.3 Induction adventitious buds Induction adventitious buds from cotyledons, it was essential that added some TDZ and NAA into induction medium. Some adventitious buds were vitrifaction, and the adventitious buds were growth normally when induced from the mediumsupplemented with TDZ 0.5 mg/l + NAA 1.0mg/l.2. The effect of different concentration NaCl on the growth of seeds and callus tissue, the main conclusion as follows:2.1 Callus tissueIn low concentration of NaCl, the growth of callus tissue kept normally level、the content of MDA was lower than check, and the activity of POD and SOD higher than check, the content of soluble sugar and Pro was higher than check. When cultured on a high NaCl concentration, the growth of callus tissue reduced rapidly, the content of MDA was increased, and the activity of POD and SOD reduced; the content of soluble sugar also reduced, but the content of Pro was still increased.2.2 SeedingsWith the increasing of NaCl concentration, the germination of seeds and activity of root system was reduced; except the seeds growth in 0.5‰NaCl, the content of seeds leave’s chlorophyll in the others treatment was lower than blank. Firstly the activity of plantlets’SOD and POD increased, subsequently decreased, and highest enzyme activity was obtain when the seeds cultured on medium contained 1‰NaCl. The content of MDA was scaled up with the increasing of NaCl concentration.更多还原