喜马拉雅红豆杉愈伤组织的诱导

采用喜马拉雅红豆杉的幼叶、幼嫩茎段、幼根为材料,比较不同外植体、基本培养基、植物生长调节剂的配比、抗氧化剂以及培养条件对愈伤组织诱导的影响。结果表明,幼叶做为外植体时,其诱导率高于幼嫩茎段和幼根,愈伤组织量大;MS、1/2MS、B5和WPM基本培养基上均能诱导出愈伤组织,其中MS基本培养基上诱导效果最好,显著高于其他的基本培养基;MS培养基上添加1 mg/L 6-BA和0.5 mg/L 2,4-D配合使用时,诱导率达到93.6%,愈伤组织生长也最旺盛;基本培养基MS中添加1 mg/L Vc时褐变现象得到极大抑制,愈伤组织生长较好;暗培养有利于愈伤组织的诱导,且诱导出的愈伤组织质量好。     

红豆杉愈伤组织诱导试验初探

以1年生红豆杉的茎段、芽尖及种子作为外植体,以MS和B5培养基添加2,4-D、NAA、KT进行愈伤组织诱导试验。结果表明:最适宜红豆杉愈伤组织诱导的条件为以茎段为外植体,消毒时间6 min;B5 NAA1.0 mg/L KT0.2 mg/L GA0.75 mg/L培养基;25℃左右培养;接种后外植体暗处理1～2周效果更佳,诱导率高,且诱导出的愈伤组织质量好。     

珙桐组织培养研究

分别以MS、N6和B5为基本培养基,添加NAA与6-BA不同浓度组合于培养基中,以珙桐的叶片、茎、芽为外植体,诱导愈伤组织。结果表明:N6培养基的诱导愈伤效果略好于MS培养基和B5培养基,但效果不显著。同时使用NAA和6-BA时,以NAA2.0 mg/L加6-BA1.0 mg/L时效果较好。叶片、茎、芽3种外植体以芽诱导愈伤组织效果最明显。     

濒危植物峨眉拟单性木兰茎段腋芽及愈伤组织诱导

以峨眉拟单性木兰带芽茎段为外植体,MS、WPM、B5为基本培养基,添加不同浓度的细胞分裂素(6-BA)和生长素(IBA、NAA、2,4-D),以不同浓度组合进行腋芽和愈伤组织的诱导.结果表明,基本培养基的类型是腋芽诱导的主要影响因素,以高盐浓度的培养基B5效果较好,MS次之,细胞分裂素的浓度与生长素的比值为15∶1时较适合腋芽的诱导,比值调整为5∶1时有利于茎的伸长生长;愈伤组织的诱导中6-BA起主导作用,其次是2,4-D,再者是基本培养基和NAA,其中2,4-D浓度过高不利于愈伤组织的诱导.     

植物生长调节剂对黑果枸杞愈伤组织诱导、增殖及分化的影响

为建立黑果枸杞高频再生体系及进行细胞突变体筛选、遗传转化奠定基础,以黑果枸杞叶片、茎段和腋芽为外植体,以MS为基本培养基,附加不同浓度的植物生长调节剂进行愈伤组织的诱导、增殖及不定芽分化研究。结果表明:诱导愈伤组织的最佳培养基为MS+2,4-D 2.0mg/L+6-BA 0.5mg/L;在愈伤组织的继代培养过程中,高浓度的2,4-D有利于愈伤组织的诱导,但对愈伤组织的生长有一定的抑制作用,最佳培养基为MS+2,4-D 1.0mg/L+6-BA 0.5mg/L;在愈伤组织的再分化过程中,6-BA对再生芽的分化效果最好,出芽率高,且多诱导出丛芽,最佳培养基为MS+6-BA 2.0mg/L+NAA 0.10mg/L。试验表明:黑果枸杞叶片、茎段和腋芽均可诱导愈伤组织,茎段诱导率最高;筛选出了茎段愈伤组织诱导、增殖及再生芽分化的适宜培养基,为其细胞工程育种及规模化生产提供支撑。     

野葛愈伤组织诱导与不定芽分化

采用MS为基本培养基，附加NAA、6-BA和IAA3种激素．诱导野葛不同外植体愈伤组织形成；再将愈伤组织接种在不同浓度6-BA的MS培养基上．分别在光照和黑暗条件下进行不定芽的诱导。结果表明．野葛幼叶、茎段和顶芽在适宜激素的诱导下均能形成愈伤组织，但不同激素组合其愈伤组织诱导率不同。幼叶愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA1．0mg/L．其诱导率为75％．茎段愈伤组织诱导的最适培养基为MS NAA1．0mg／L 6-BA3．0mg／L IAA0．2mg/L，其诱导率为70％。不同外植体形成的愈伤组织其不定芽的分化诱导率不同．茎段愈伤组织在不同浓度6-BA下均能诱导出不定芽；光照有利于芽的分化．在光培养条件下，茎段愈伤组织不定芽平均诱导率为66．7％。     

丝棉木松散型胚性愈伤组织的诱导与增殖

通过建立丝棉木松散型胚性愈伤组织培养体系,为丝棉木组培快繁、体细胞离体诱变育种及遗传转化等研究奠定基础。本试验以丝棉木幼嫩茎段为外植体,研究植物生长调节剂不同种类和浓度对丝棉木松散型胚性愈伤组织诱导与增殖的影响。结果显示,丝棉木茎段在MS+1.0 mg/L 2,4-D培养基上可诱导出松散型愈伤组织,愈伤组织诱导率达100%;将松散型愈伤组织转接到MS+0.5 mg/L BA+0.5 mg/L 2,4-D的培养基上可获得胚性愈伤组织;将胚性愈伤组织转接到MS+0.25 mg/L BA+0.5 mg/L 2,4-D的培养基上,每15 d继代培养一次,继代时选择结构松散、质地硬脆的愈伤组织,继代培养2～3次后可获得均质的丝棉木松散型胚性愈伤组织。本研究表明,丝棉木茎段理想的松散型愈伤组织诱导培养基为MS+1.0 mg/L 2,4-D;最适松散型愈伤组织诱导培养基为MS+0.5 mg/L BA+0.5 mg/L 2,4-D;最适胚性愈伤组织增殖培养基为MS+0.25 mg/L BA+0.5 mg/L 2,4-D。     

针叶十大功劳愈伤组织诱导研究

针叶十大功劳Mahonia fortunei生长期长、市场需求量大、野生资源日益匮乏的现状,首次以针叶十大功劳Folium Mahoniae茎、叶为外植体诱导愈伤组织生长.实验结果表明选用B5及MS培养基,温度24±1℃,暗培养条件下均可诱导出愈伤组织,但B5培养基更适宜于针对十大功劳愈伤组织诱导.培养基中添加植物激素2,4-D0.5 mg/L、KT0.5 mg/L时,愈伤组织诱导率最大,且外植体污染率最低.相同培养条件下,选用茎为外植体较选用叶片为外植体可获得更高的愈伤组织诱导率.培养基中添加3%蔗糖有利于愈伤组织的诱导,但添加NH4NO3则可明显降低愈伤组织诱导率.研究结果对开展十大功劳的人工繁殖研究具有一定的参考价值.     

芍药茎段愈伤组织诱导的研究

以芍药品种‘粉玉奴’茎段为实验材料,1/2 MS培养基为基本培养基,探究不同浓度的6-BA以及不同光质对芍药茎段愈伤组织诱导的影响。试验结果表明,6-BA浓度为0.5 mg/L时芍药茎段愈伤组织诱导情况最好;最适宜芍药茎段愈伤诱导的培养基为:1/2MS+Ca~(2+)(WPM)+PVP 1.0 g/L+NAA 1 mg/L+LH 0.5g/L+蔗糖30 g/L+琼脂7 g/L+6-BA 0.5 mg/L;不同的光质条件下,红光更有利于芍药进行愈伤组织诱导。     

广藿香愈伤组织的诱导和增殖

以广藿香的幼叶、茎段为外植体,接种于附加不同浓度的2,4-D、NAA、6-BA、KT、TDZ及其组合的MS固体培养基上进行愈伤组织的诱导,结果表明:单独使用5种激素在一定范围内均能诱导出愈伤组织;最佳组合培养基为MS+6-BA 1.0 mg.L-1+2,4-D 0.1 mg.L-1。进一步研究发现,茎段和叶片诱导的愈伤组织的生长曲线都呈“S”型,且生长周期均为18 d。     

虎杖愈伤组织的诱导及其白藜芦醇形成初探

将虎杖P olygonum cusp id a tum不同外植体经不同消毒时间处理后,接种在添加不同激素种类和水平的相同基本培养基上或相同激素种类和水平基本培养基上进行诱导实验,同时对根和根茎芽、叶、韧皮诱导的愈伤组织进行白藜芦醇含量的测定.结果表明:基本培养基以M S较好,外植体叶对激素种类较为敏感,其中适当浓度的NAA诱导愈伤组织比2,4-D的效果好,KT比BA好,添加KT的培养基上诱导愈伤组织比较紧密,有利于分化,在M S+NAA 2 m g/L+KT 0.1 m g/L培养基上诱导愈伤组织较好,根茎芽的诱导率最高,为73%;愈伤组织的生长趋势从接种的第3天开始生长,到21 d时生长达到最高峰,干质量为0.461 2 g,以后生长速度减慢;对不同材料诱导的愈伤组织进行白藜芦醇含量的测定,其中根茎部芽的诱导的愈伤组织中白藜芦醇含量最高,其次是叶和根,最低的为韧皮.     

Establishment of cell suspension line of Populus tomentosa Carr

Leaves of fine Populus tomentosa genotype TC152 were used as explants to establish cell suspension lines. The effects of plant growth regulators on callus induction and establishment of cell suspension lines were studied. The callus induction rate was the highest on a MS solid medium supplemented with 1.0 mg·L-1 2,4-D. A cell suspension line could be obtained by inoculating calli which were not subcultured into a MS liquid medium supplemented with 1.5 mg·L-1 2,4-D. The best subculture medium was MS + 0.8 mg...     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ＇Moldova＇

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Somatic embryogenesis and histological analysis from zygotic embryos in Vitis vinifera L. ''Moldova''

We examined the somatic embryogenesis from and histological studies of zygotic embryos of seeds in European Grape 'Moldova' (Vitis vinifera L. 'Moldova'). Primary calli were initiated on Nitsch and Nitsch (NN) medium supplemented with 1.0mg'L-1 2,4-D and 0.5 mg'L-1 6-BA. Embryogenic calli were produced upon transfer to a NN medium with 0.5 mg-L-1 6-BA and 2 mg.L-1 NAA and somatic embryos were obtained on a half strength MS medium without plant growth regulators. During the somatic embryo germination, an addition of 1.0 mg.L-1 6-BA in the medium could accelerate somatic embryos to develop into normal plants and increase the conversion rate from 0 to 43.3%. Histological studies of embryogenic calli and somatic embryos demonstrated dy-namic changes of proteins and starch grains. The developmental processes of somatic embryos were similar to those of zygotic em-bryos, including typical epiderma, cotyledon primordium and vascular tissue.     

Efficient embryogenic callus induction and plant regeneration from embryonic axis explants inQuercus acutissima

Embryogenic callus ofQuercus acutissima was successfully induced from embryogenic cultures, and plants were regenerated from the callus. The development of the techniques involved will allow mass propagation and gene transformation in this species. Embryogenic cultures were formed from embryonic axis explants (i.e., embryos without cotyledons) excised from immature embryos, after culture on Murashige and Skoog (MS) medium containing indolebutyric acid and benzyladenine. Attempts to induce embryogenic cultures from cotyledon explants were unsuccessful. Embryogenic calli were induced at high frequency from embryogenic cultures on MS medium containing 2,4-dichlorophenoxyacetic acid. However, benzyladenine inhibited embryogenic callus formation. Somatic embryo development from embryogenic calli occurred on MS medium in all of the seven cell lines tested. Germination of somatic embryos was induced on half strength MS medium without plant growth regulators. Finally, acclimated plants growing in soil were obtained.     

Somatic embryogenesis and plant regeneration inAcanthopanax sciadophylloides

Somatic embryos ofAcanthopanax sciadophylloides Franch. et Sav. were differentiated from both zygotic and somatic embryos and calli, and plants were regenerated from these somatic embryos. A zygotic embryo, enclosed within a small portion of the endosperm, was incubated on Murashige and Skoog (MS) media supplemented with various combinations (range 0–10.0 mg/l) of 6-benzylaminopurine (BAP) and 2,4-dichlorophenoxyacetic acid (2,4-D). After 4 months, swelling of the zygotic embryos and callus formation was observed. When the swollen embryos were transferred to MS medium supplemented with 0.5 mg/l of 2,4-D, somatic embryos were formed in one to two months. After subculture on the same medium, new embryos were differentiated from various parts of the older somatic embryos. The calli were cultured on medium supplemented with 2.0 mg/l of 2,4-D and BAP for three weeks. Proliferated calli were transferred to medium supplemented with 1.0 mg/l of 2,4-D and BAP. Somatic embryos were differentiated from the calli within one to two months. Somatic embryos were germinated on half-strength MS medium without plant growth regulators and the plantlets were grown in soil. A part of this paper was presented at the 106th Annual Meeting of the Japanese Forestry Society (1995) & First Asia-Pacific Symposium on Forest Tree Genetic Improvement (Beijing).     

盾叶薯蓣愈伤组织培养及其高产系的筛选

以盾叶薯蓣的块茎、茎、叶和茎尖为材料,对其进行了愈伤组织培养,并根据愈伤组织个体间生长速度和皂甙含量差异,进行了高产系的筛选．结果表明：盾叶薯蓣不同外植体（块茎、茎、叶、茎尖）均能诱导出愈伤组织,其中以茎尖形成愈伤组织最快,皂甙元含量最高;不同培养基、ｐＨ 值、接种量、温度、激素等因子对愈伤组织的形成、生长及皂甙元含量有很大的影响;改良 Ｍ Ｓ培养基、２,４－Ｄ 浓度为２．０～４．０ ｍ ｇ／Ｌ、ＢＡ 为 ０．５ ｍ ｇ／Ｌ 的激素配比使愈伤组织的生长量、产物含量最高;对无性系进行筛选和 ＨＰＬＣ 检测,以 １２号无性系生长速率和皂甙元含量最高,其皂甙含量达 ３．７％ ,生长量比普通系高２１．５％ ．     

Somatic embryogenesis from cell suspension cultures of aspen clone

Suspension cultures initiated from callus derived from petiole explants of aspen hybrid (Populus tremuloides × P. tremula) produced somatic embryos. Callus was induced on a MS medium supplemented with 5 mg·L–1 2,4-D and 0.05 mg·L–1 zeatin under light conditions. Embryogenic calli were obtained when a subsequent subculture of calli was suspended in the same basal me-dium with 10 mg·L–1 2,4-D. The highest number of globular embryos were induced from embryogenic calli by cell suspension cul-ture in a MS liquid medium supplemented with 10 mg·L–1 2,4-D. Genotype and 2,4-D concentration were vital to the induction of embryogenic calli producing competent cells. Embryogenic calli for each genotype were heterogeneous. Green calli with gel-like consistency could yield more competent cells than light yellow embryogenic calli. However, some globular embryos broke into slices and some developed abnormally after one month of culture under the same or other hormonal conditions.     

In vitro plant regeneration of Eclipta alba and increased production of coumestans

Eclipta alba plants were regenerated from totipotent calli of leaf, stem and root cultured on Murashige and Skoog’s medium supplemented with various growth regulators. The production of the coumestans wedelolactone and demethylwedelolactone was estimated by UV method.