基于不同激素组合的紫丁香成熟种子组培诱导研究

以紫丁香成熟种子为外植体,采用不同激素组合培养基进行组培诱导试验的结果表明:采用中间切割方式的种子萌发率较高,最高可达59.72%;NAA与BA组合诱导丁香种子效果较好,NAA0.1 mg/L+BA1.0 mg/L的诱导率可达88.89%,MS+BA3.0 mg/L为顶芽和带芽茎段的最适增殖继代培养基;茎芽在试管内、外生根时间分别为20天和4-6周,生根率均为100%,紫丁香成熟种子组织培养试验取得成功。     

珍稀竹种巨龙竹组织培养研究

对巨龙竹幼年竹和成年竹材料采集、处理、试管引入、丛芽诱导、丛芽增殖和生根、再生植株炼苗等进行系统研究,采用正交设计筛选出各阶段的最佳培养基.结果表明:丛芽增殖倍数与BA用量成正比,但随着BA用量加大或长期在高浓度BA上培养易产生花芽,说明BA有使丛芽细胞从营养状态向生殖状态转变的作用;KT在丛芽高生长方面起主导作用,能使丛芽节间伸长,使细胞从生殖状态向营养状态转变;椰乳有改善丛芽或苗生长的作用.在诱导丛芽生根过程中,IBA起主导作用,配合少量NAA使用可得到生根良好、移栽成活率较高的再生小植株.巨龙竹成年竹组织培养与外植体采集部位、采集时间、采集地等相关.     

In vitro regeneration of plantlets from mature embryos of Pinus ayacahuite

A plantlet regeneration protocol was developed for Pinus ayacahuite var. ayacahuite (Ehrenb.). Embryos from mature seeds from ten provenances were cultured in a 16-h photoperiod for 3 days on a medium containing 30 mM sucrose and 0.7% agar. Cotyledons from these embryos were subcultured onto MCM medium (Bornman 1983) supplemented with 50 micro M N(6)-benzyladenine and 90 mM sucrose for 2 weeks. Bud development and shoot elongation were maximized by subculturing the explants on half strength AE medium (von Arnold and Ericksson 1981), supplemented with 60 mM sucrose and 0.05% activated charcoal every 30 days. Seed source had a significant effect on the responses of the embryos to the bud induction protocol. For the provenance with the best response to bud induction, about 79% of the cultured cotyledons formed buds, and each cotyledon formed a mean of 9.1 buds, so that about 70 shoots could be induced from each seed. The best rooting response (40% rooting) was obtained by treating the shoots for 8 h with 100 micro M naphthalene acetic acid.     

Rapid clonal multiplication of a woody tree, <Emphasis Type="Italic">Vitex negundo</Emphasis> L. through axillary shoots proliferation

An efficient and improved shoot regeneration technique for the micropropagation of Vitex negundo, an aromatic and medicinal shrub through in vitro culture of nodal segments with axillary buds, is described. Thidiazuron (TDZ) used at 1.0 μM was the most effective in inducing bud break and growth, and also in initiating multiple shoot proliferation at the rate of 25 microshoots per nodal explant with axillary buds, after 4 weeks of culture. By repeated subculturing of nodal explants, a high-frequency multiplication rate was established. Optimum shoot multiplication and elongation was achieved when TDZ exposed explants were subcultured on Murashige and Skoog (MS) media containing a combination of 1.0 μM 6-benzyladenine (BA) and 0.5 μM α-naphthalene acetic acid (NAA). Efficient rooting was achieved directly in soilrite when basal portion of the shoots were treated with 500 μM indole-3-butyric acid for 10 min which was the most effective in inducing roots, as 97% of the microshoots produced roots. Plantlets went through a hardening phase in a controlled plant growth chamber, prior to ex-vitro transfer. Micropropagated plants grew well, attained maturity and flowered. No phenotypical differences for morphogenesis were observed among the regenerants.     

西蒙得木组织培养繁殖技术的研究

通过对西蒙得木〔Simmondsia chinensis(Link)〕组织培养大批量繁殖技术的研究,探索基本培养基、生长调节物质及蔗糖含量等对嫩梢增殖及生根的影响,结果表明:采用春季生长的1年生实生苗以及6年生高产的优株茎节作外植体,培养于含有ZT 1～3 mg/L或BA 1～3mg/L和NAA 0.01～0.1 mg/L的MS培养基中,经30～40天,几乎所有腋芽均能长成嫩梢,切取长达4～6 cm的嫩梢,培养于含有NAA的1/2MS培养基中,经1周暗培养,1个月后生根率达93％,小植株移栽于含砂土的基质中,保持湿润,获得成功。用幼苗及优株作外植体的培养效果无显著差异,说明此法不受遗传型的限制。     

金冠白蜡快繁体系建立的研究

以金冠白蜡带腋芽的茎段作为外植体,进行组织培养快繁体系的建立。结果表明:使用消毒剂0.1%HgCl2+吐温消毒3min为最佳外植体消毒处理,存活率可达88.3%;启动培养基采用MS+6-BA 1.0mg/L+IBA 0.2mg/L为宜;继代增殖培养选择MS+6-BA 2.0mg/L+KT 1.0mg/L+IBA 0.5mg/L的培养基,其增殖系数为4.617;生根培养以1/4MS+IBA 1.0mg/L的培养基进行培养,生根率达75%。     

孔雀草杂交F1代的植株再生

以孔雀草（Tagetes patula）子叶、下胚轴和叶片为外植体,通过器官直接发生途径诱导形成不定芽,探讨植物生长调节剂组合、AgNO3浓度、蔗糖浓度和外植体类型等因素对植株再生的影响,建立了再生体系。结果表明：MS＋6-BA 1.0 · L^- 1＋NAA 0.5 · L^- 1＋AgNO31.0 · L^- 1＋蔗糖40 g·L^-1培养基最适合不定芽的分化和增殖,子叶不定芽分化率达90%以上,平均每外植体分化不定芽数达5.3个。不定芽较适生根培养基为1/2MS＋IAA 0.2 · L^- 1＋NAA 0.1 · L^- 1,生根率达到90%。     

In vitro regeneration of Pacific silver fir (Abies amabilis) plantlets and histological analysis of shoot formation

A protocol is described for the in vitro propagation of Abies amabilis (Dougl.) Forbes. Over 60% of the cotyledonary explants from 5-day-old cotyledons formed shoots when cultured for 7 days on Schenk and Hildebrandt's (SH) medium containing 10 micro M N(6)-benzyladenine (BA) followed by another 7 days on SH medium containing 10 micro M each of BA and zeatin. Shoot multiplication was unsuccessful. Seventeen percent rooting was obtained after pulsing for 4 h in 1 mM indole-3-butyric acid and planting pulsed shoots in 1/1 (v/v) perlite/vermiculite. Cell clusters (promeristemoids) of five to seven cells were observed on the cotyledonary explants after 7 days of culture in the presence of cytokinin. These cells developed further into meristematic domes and apical meristems. In the absence of cytokinin, stomata and resin canals reached maturity, whereas cells within the cortex became vacuolated and developed into palisade and spongy mesophyll.     

Plant regeneration in <Emphasis Type="Italic">Actinidia polygama</Emphasis> Miq. by leaf,stem, and petiole culture with zeatin,and from stem-derived calli on low-sucrose medium

We examined the effect of zeatin on the formation of shoot buds from explants and callus tissues derived from stem segments of Actinidia polygama Miq. (matatabi or silver vine). Stem and petiole segments cultured on combined broad-leaved tree medium and woody plant medium (BW medium) supplemented with zeatin for 40 days formed many shoot buds. Callus tissues derived from the stem segments and subcultured on BW medium supplemented with 6-benzyladenine and 1-naphthaleneacetic acid formed shoot buds when the medium contained 13.7µM zeatin. BW medium containing low concentrations of sucrose strongly induced the formation of shoot buds from the callus.     

菊花脱除花叶病毒组培技术研究

以患花叶病菊花0.5~0.8 mm幼嫩茎尖为外植体,愈伤组织的诱导在MS+BA2.0mg/L+NAA0.1 mg/L培养基上效果最好,诱导率达96.0%;愈伤组织分化出芽以MS+BA3.0 mg/L+NAA0.1 mg/L效果最佳,分化率达60.0%,每块愈伤组织平均分化出4个芽;幼苗在1/2MS+NAA0.1 mg/L培养基中生根率达92.0%,株平均生根5.1条。组培苗经症状观察和汁液传染实验试验检测,以后种方法结果为依据,获得菊花脱毒苗19株,脱毒率为63.3%。     

荻植株再生体系建立

以成熟种子为外植体建立了荻植株再生体系。荻成熟种子经20％“84”消毒液处理20min,接种于MS＋4％蔗糖（pH5．5）培养基中培养5d,萌发率为92％,褐化率为8％,污染率为0。萌发种子继代转入愈伤诱导培养基MS＋lmg／16-BA＋1mg／12,4-D＋0．5mg／lNAA＋4％蔗糖（pH5．5）,光照条件下愈伤组织诱导率为86％,黑暗条件下愈伤组织诱导率64％。荻愈伤组织在Ms＋1IYlg,L6-BA＋0．1mg／L2,4-D＋0．5mg／LNAA＋4％蔗糖（pH5．5）进行不定芽诱导,不定芽发生率为91％。无根丛生芽继代转入不定根诱导培养1／2MS＋0．25mg／LIBA＋0．25mg／LNAA＋4％蔗糖（pH5．5）,5天可见不定根形成,15天后不定根发生率为92％。研究还发现,荻种子萌发后可直接继代转入不定芽分化培养基形成健壮不定芽,不定芽诱导率为95％。     

Caulogenic callus induction and adventitious bud formation from embryos of long-term stored seeds ofPicea jezoensis

Caulogenic calli with a high differentiation potency were induced from mature embryos ofPicea jezoensis seeds stored over a long time, for 29 years, resulting in the active formation of adventitious buds. Embryos began to induce calli within 3 weeks of cultivating on LP medium containing 3 μM BAP and 1 μM 2,4-D. Then, the calli proliferated and transformed into caulogenic calli with bud primordia in 8 weeks. The caulogenic calli increased actively with the addition of 500 mg/l ofl-glutamine in the medium. Furthermore, caulogenic calli, induced on LP medium containingl-glutamine, resulted in the formation of adventitious buds, which elongated after transferring the calli into LP medium with 0.1 μM BAP, but withoutl-glutamine. It appears that the number of adventitious buds and the process of shoot elongation are influenced by the kind of nitrogen contained in the medium for callus induction. A part of this study was presented at the 107th Annual Meeting of the Japanese Forestry Society (1996).     

细胞分裂素和硝酸铵对欧洲云杉不定芽伸长的影响

报道细胞分裂素和硝酸铵对欧洲云杉不定芽伸长过程的影响.采用5年生的欧洲云杉1个无性系不定芽丛来研究玉米素和硝酸铵对不定芽生长的影响.试验3种诱导类型(玉米素浓度分别为6.84、13.68、20.52 μmol·L-1,激动素浓度0.69 μmol·L-1,培养4周),玉米素最高浓度处理诱导的伸长芽数量最多,在4个无生长调节剂继代培养过程中,加入了1个硝酸铵浓度逐渐增加的继代培养过程.经不含生长调节剂的低浓度的硝酸铵(2.5 mmol·L-1)培养基的继代培养后转入含有细胞分裂素的诱导培养基,下一个继代培养基补充的硝酸铵浓度分别为2.5、7.5、12.5、17.5 mmol·L-1,8周(2个继代培养)后转入含有硝酸铵浓度为2.5 mmol·L-1的培养基培养.硝酸铵浓度为7.5 mmol·L-1的处理可形成数量最多的不定芽,这对于采用不同浓度的玉米素进行诱导的处理结果是一致的,但随着硝酸铵的浓度增至12.5 mmol·L-1和17.5 mmol·L-1时则不定芽的数量减少.欧洲云杉的胚轴外植体经1次或多次诱导后,细胞分裂素(BA、激动素、玉米素、2iP)促进了不定芽的分化和发育,其中BA由于可诱导产生大量的不定芽而成为最有效的细胞分裂素.玉米素虽然诱导不定芽相对弱一些,却在BA多次诱导后可促进芽的伸长.这对获得具有形成不定芽能力并可扩繁的芽丛非常重要.     

外植体及培养基对欧洲白桦器官再生的影响

白桦再生不定芽以通过愈伤组织再生不定芽的间接方式为主,无菌试管苗的茎段、根、叶片均可作外植体诱导不定芽再生。不同的基因型其不定芽再生的难易程度不一样。无性系2/86比G1、4N容易诱导愈伤组织进而诱导不定芽发生。诱导无性系2/86茎段不定芽分化的适宜基本培养基为WPM,根段为MCM,叶片为MS。不同外植体不定芽再生能力也不同,叶柄和茎段最容易诱导愈伤组织和不定芽分化,根较难诱导。     

光皮树的组织培养研究初探

以光皮树种子和嫩枝条为材料,探讨不同诱导途径对培养的影响:①分别以茎段、顶芽、叶片、带腋芽的茎段及无菌发芽的种子苗上的胚轴和子叶为外植体离体诱导出愈伤组织,然后再进行分化;②以带腋芽的茎段为外植体进行腋芽增殖.结果表明:70％酒精与0.1％升汞配合使用,消毒效果较好.其中,茎段消毒采用70％酒精浸泡30 s,0.1％升...     

Micropropagation of red sanders (Pterocarpus santalinus L.) using mature nodal explants

In vitro clonal multiplication of Pterocarpus santalinus L. was achieved using mature nodal explants of a 10-year-old elite quality tree. Combinations of serial transfer technique and incorporation of antioxidants (250 mg/l L-ascorbic acid and 50 mg/l citric acid) into the culture medium helped to minimize medium browning and improve explant survival during shoot sprouting. About 70% of explants were sprouted on Murashige and Skoog (MS) liquid medium containing 4.4 μM 6-benzyladenine (BA). The explant harvest period also influenced the bud break and shoot sprouting in nodal explants. The combination of 4.4 μM BA and 2.2 μM thidiazuron (TDZ) was found to be the most suitable growth regulator for obtaining the highest percentage of nodal segment sprouting (74%–75%), the number of secondary shoots per primary shoot (two or three), the shoot length (5–6 cm), the number of new nodal segments generated per active explant (four or five), and the multiplication coefficient (3.5) within 6 weeks. Repeated subculturing of nodal explants obtained from shoot cultures enabled continuous production of healthy axillary shoots. At the end of the sixth passage, about 90% of nodal explants produced five or six healthy green shoots, each being about 6.6 cm long with six or seven nodes. Multiplication coefficient was also increased from the first subculture (5.4) to the sixth subculture (8.3). The best rooting response was achieved on solidified half-strength MS medium supplemented with 4.9 μM indole-3-butyric acid (IBA). About 70% of the micropropagated plantlets were established successfully in 20-cm pots containing a mixture of soil and farmyard manure (4 : 1 ratio) and formed new leaflets.     

A regeneration system using cotyledons and cotyledonary node explants of Toona ciliata

We used the cotyledons and cotyledonary nodes of Toona ciliata(Chinese mahogany) as explants to examine callus and adventitious shoot induction when exposed to different ratios of hormones. We also investigated the effects of seedling age, inoculation method, and genotype on the efficient regeneration of T. ciliata. The results showed that different genotypes exhibited significantly different callus induction efficiency. The cotyledons and cotyledonary nodes of 20-day seedlings inoculated onto MS medium with 0.5 mg/L 6-benzylaminopurine(6-BA), 0.5 mg/L kinetin(KT) and 0.05 mg/L 1-naphthylacetic acid(NAA) achieved a greater regeneration rate than did other concentrations of cytokinin and auxin. The numbers of shoots per cotyledon and cotyledonary node explant were 7.33 and 6.67. The optimal inoculation method for cotyledons was that the distal end of the explants was placed in contact with the medium. The optimal adventitious shoot differentiation medium for cotyledon explants was MS medium containing 0.3 mg/L 6-BA and 0.2 mg/L NAA, producing a 3.4 cm height of shoot on average. This study established an efficient regeneration system for T. ciliata with cotyledons and cotyledonary nodes as explants.     

茛艻花组织培养技术

以茛艻花顶芽为外植体进行离体培养,成功建立了快速繁殖技术体系,结果表明,不同激素及其浓度对其增殖及根的形成影响显著。各个阶段适宜培养基:(1)增殖培养基为MS+BA 3.0 mg/L+NAA 1.0 mg/L,培养30天,增殖率稳定为4.65;(2)壮苗培养基为MS+6-BA 0.2 mg/L+IBA 0.2 mg/L;(3)生根培养基为1/2MS+NAA4mg/L+蔗糖20 g/L时,生根率达95%。     

不同部位银杏茎段培养及位置效应的研究

以一个月月龄银杏(Ginkgo biloba)无菌幼苗各部位茎段为外植体,研究其顶芽、带腋芽的中段、带子叶的下段腋芽萌发和不定芽的分化情况。结果表明:①银杏顶芽段(上段)在以改良MS、N6、DCR为基本培养基,附加0.1mg/LNAA和0.5mg/L6-BA的培养基上顶芽生长较其他培养基好,抽出的芽苗长得高,最高达2.98cm,未见有不定芽的分化;②中段外植体在四种基本培养基(MS、DCR、N6、改良MS)中附加0.1mg/LNAA和0.5mg/L6-BA和0.25%AC时,都能诱导出腋芽和不定芽,4种培养基之间芽诱导率无明显差异,以N6培养基的诱导效果最好,平均每个外植体产生1.56个芽;③带有子叶下段的外植体培养后,能诱导出2～3个子叶腋芽和新的不定芽,在改良MS+0.1mg/LNAA+0.5mg/L6-BA、N6+0.1mg/LNAA+0.5mg/L6-BA培养基上两个腋芽的诱导率均达80%以上。表明,不同部位的银杏茎段在芽的诱导上存在位置效应,可利用不同部位进行银杏的定向快速繁殖。