Beneficial effects of activated charcoal on bulblet production in tissue cultures of Muscari armeniacum

Production of bulblets of Muscari armeniacum through tissue culture is enhanced when 1 g/l activated charcoal is added to a modified Murashige and Skoog (MS) medium. Bulblet regeneration is direct from bulb scale explants with no intermediate callus growth. Bulblets can be transferred successfully to a greenhouse environment directly from aseptic culture.     

Efficient <Emphasis Type="Italic">in vitro</Emphasis> bulblet regeneration from immature embryos of endangered <Emphasis Type="Italic">Sternbergia fischeriana</Emphasis>

Sternbergia fischeriana is an endangered geophyte and therefore in vitro micropropagation of this plant will have great importance for germplasm conservation and commercial production. Bulb scale and immature embryo explants of S. fischeriana were cultured on different nutrient media supplemented with various concentrations of plant growth regulators. Immature embryos produced higher number of bulblets than bulb scales. Large numbers of bulblets were regenerated (over 80 bulblets/explants) from immature embryos on Murashige and Skoog (MS) medium supplemented with 4 mg l^–1 6-benzylaminopurine (BA) and 0.25 mg l^–1 -naphthaleneacetic (NAA) or 2 mg l^–12,4-dichlorophenoxyacetic acid (2,4-D) after 14 months of culture initiation. Regenerated bulblets were kept at 5 °C for 5 weeks and then transplanted to a potting mixture.     

太仓大蒜根尖离体培养直接诱导不定芽及其试管鳞茎的形成

用正交设计法研究6-BA和NAA及根尖长度对太仓大蒜根尖不定芽直接诱导的影响,结果表明,三者都有极显著影响,6-BA影响最大,NAA次之,根尖长度最小;筛选出的最佳生长调节物质组合为6-BA 3～5mg·L-1 NAA1.0mg·L-1,切取的最佳根尖长度为1.2mm.并用得到的试管苗成功诱导形成了试管鳞茎,质量在60～320mg之间,破除休眠后,播种在蛭石和珍珠岩(3:1)混合的基质上,出苗率为91.3%,长势良好.     

Adventitious shoot regeneration from leaf explants and stem nodes of Lilium

A method for the regeneration of lily plantlets (Lilium spp.) through different morphogenic pathways is described. Plant regeneration was obtained from in vitro cultured leaves of four lily hybrids, cultured on Murashige and Skoog's basal medium supplemented with cytokinins (TDZ and BA) and auxins (NAA and IBA) at different concentrations. Direct shoot regeneration occurred with all tested media for the Asiatic lilies `Elite' and `Pollyanna' and also for the Oriental hybrid `Star Gazer'. Callus developed on TDZ-enriched medium from leaf segments of L. longiflorum cv. `Snow Queen' regenerated by direct organogenesis. This occurred on a medium with auxin/ cytokinin balance which was lower than other genotypes. There were fewer problems of sterilization with leaves from sprouted bulbs than in vitro scale culture. This suggests that the leaf-segments obtained in this way could be an alternative to the scales as a source of material for propagation. A protocol for micropropagation based on bulblets from in vitro shoot-tip-derived stem nodes was also used. The development of pseudo-bulbets is particularly advantageous since it allows for structures characterised by absent or low dormancy. Regenerated shoots have been rooted and successfully acclimatized to greenhouse conditions where they flowered after the second year giving plants with true-to-type shape and colour.     

兰州百合组培鳞茎发育研究

该研究以兰州百合商品种球鳞片为外植体材料,通过组织培养诱导丛生芽萌发及高频增殖,再以丛生芽为材料诱导其发育形成小鳞茎,调节培养基对小鳞茎进行膨大发育培养,最终形成促进兰州百合组培鳞茎膨大发育的“三步”组培培养技术路线;对发育过程中形成的丛生芽、小鳞茎及膨大鳞茎进行淀粉含量测定与生长特征参数统计,分析各步培养对鳞茎形成发育过程中淀粉含量与形态变化的影响。结果表明：所建立的“三步”培养方案培育出的组培鳞茎直径、重量与鳞片数分别为1.66 cm、2.48 g和26.33片,有效地促进了鳞茎的膨大,并能诱导鳞茎主茎杆的形成发育;在培养进程中其淀粉含量呈现逐步增加的趋势,这表明与鳞茎膨大发育正相关,同时鳞茎大小、重量及鳞片数三者也表现为正相关性;当鳞茎所含鳞片数在26片以上时,其生长点易发育形成主茎杆。该文研究了兰州百合组培鳞茎的形成与膨大发育技术,所研发的“三步”培养组成的鳞茎膨大发育组培技术有效地促进了鳞茎的膨大发育,而膨大发育的鳞茎能有效地缩短田间生长周期,从时间上提高百合生产量,同时为实现兰州百合膨大的鳞茎种球规模化生产提供技术支撑。     

In vitro propagation of red squill,Urginea maritima Baker

Red squill (Urginea maritima Baker), a plant in the Liliaceae of potential use as a rodenticide, was successfully propagated in vitro. Bulblets were induced from red squill bulb-scales cultured in the dark on a medium containing Murashige and Skoog [6] salts supplemented with a combination of 0.5 or 1.6 M naphthaleneacetic acid and 0.4 or 1.3 M 6-benzylaminopurine. Bulblets induced in vitro were rooted in medium containing 0.5 or 1.6 M naphthaleneacetic acid and planted in vermiculite. Bulb-scale explants could be returned to fresh medium regenerate new bulblets, which were easily rooted. Alternatively, bulblets could be subcultured to regenerate adventitious shoots in medium containing 4.4 or 13.2 M 6-benzylaminopurine. Shoots regenerated in this manner proved difficult to root by a number of treatments tested. Regenerated bulblets were chilled at 5C for 3 to 4 weeks to induce leaf emergence prior to transfer to the greenhouse.     

百合鳞茎发育过程中淀粉合成相关酶基因的克隆及表达分析

该研究通过同源克隆技术克隆腺苷二磷酸葡萄糖焦磷酸化酶(AGPase)、颗粒结合淀粉合酶(GBSS)和可溶性淀粉合酶(SSS) 3类百合淀粉合成关键酶基因,分析这三类淀粉合成关键酶基因的表达变化,测定百合鳞茎膨大发育中淀粉含量变化。结果表明:(1) AGPase具有GlgC家族蛋白PLN02241蛋白结构特征及cl11394家族蛋白ADP_Glucose_PP与NTP_transferase结构域,获登录号KP751443; GBSS与SSS具有cl10013家族蛋白Glyco_transf_5,GT1_Glycogen_synthase_DULL1_like结构域,获登录号分别为KP751444、KP751445。(2)百合鳞茎形成与膨大发育过程中,淀粉含量呈现递增趋势,鳞茎盘开始分化茎杆时其淀粉含量最高,达到44.52%。鳞茎与叶片部位的三个淀粉合成相关酶基因表达量均逐渐增加;在鳞茎膨大后茎杆分化阶段,三个淀粉合成相关酶基因表达量达到最高,AGPase、GBSS、SSS在鳞片中的表达量分别为10.79,6.92和5.12,叶片中的表达量分别为6.79,5.22和4.41,鳞片中的表达量大幅度高于叶片;淀粉合成相关酶基因的表达量变化与淀粉含量、鳞茎的膨大发育成正相关。这为鳞茎的繁殖生产提供了可通过调节淀粉合成关键酶基因表达促进百合鳞茎膨大发育的思路。     

温度对兰州百合鳞片埋培繁殖的影响

为了筛选出兰州百合鳞片埋培繁殖的最适温度和鳞片层次,解决兰州百合种源不足、繁育周期长的问题,该文以兰州百合鳞片为材料,采用温度(20、25、30℃)和鳞片层次(外层、中层、内层)二因素完全随机区组设计,研究了二因素对兰州百合鳞片埋培繁殖效果的影响。通过对鳞片疑似发病率、分化率及小鳞茎分化数进行统计与分析,结果发现不同温度处理及各鳞片层次对鳞片疑似发病率、分化率及小鳞茎分化数的影响存在显著或极显著差异。结果表明:(1)温度越高,鳞片的疑似发病率越低,在埋培2周时,20℃处理下疑似发病率最高(38.67%),30℃处理下最低(10%);各层次鳞片的疑似发病率由高到低依次为外层中层内层。(2)在25、30℃处理下,小鳞茎分化率最高,埋培结束(6周)时分别为91.33%、90.89%;中层及内层鳞片小鳞茎分化率极显著高于外层鳞片。(3) 30℃处理下鳞片形成小鳞茎数最多,在埋培6周时达到每片2.00粒;同时中层及内层鳞片小鳞茎分化数显著高于外层鳞片。综上结果表明,兰州百合鳞片埋培繁殖以选用中层(3~4层)、内层(5~7层)鳞片在25~30℃条件下繁殖效果最好。     

Manipulation of the morphogenetic pathways of Lilium longiflorum transverse thin cell layer explants by auxin and cytokinin

Summary Excised tissues from transverse young stem sections of Lilium longiflorum were cultured on Murashige and Skoog medium supplemented with growth regulators at various concentrations. After 45 d in culture, the presence of α-naphthaleneacetic acid (NAA) in the culture medium at 5.4 μM resulted in bulblet formation while 2,4-dichlorophenoxyacetic acid (2,4-D) at 2.2 μM resulted in root formation. The presence of IBA (indole-3-butyric acid) in the culture medium at 1.0 μM resulted in shoot formation while plantlet formation occurred when IBA was added at a concentration of 2.0 μM. When 1-phenyl-3-(1,2,3-thiadiazol-5-yl)-urea (TDZ) was added to the culture medium at 1.1 μM, protocorm-like bodies (PLBs) formed, while 2.2 μM resulted in shoot formation (on abaxial and adaxial surfaces). The presence of NAA and TDZ in the culture medium at 5.4 μM and 0.4, 1.1 or 2.2 μM, respectively, resulted in somatic embryo formation while NAA- and 6-benzylaminopurine-(BA) containing culture medium formed callus or bulblets. The establishment of different regeneration systems when explants are exposed to various growth regulators demonstrates that the choice of growth regulator combinations and concentrations are of significance in determining the morphogenetic response and plant regeneration capacity.