大豆体细胞胚胎发生影响因素的研究

研究了影响大豆幼胚培养体细胞胚胎发生频率的9个因素。诱导体细胞胚胎发生的适宜幼胚长度为4mm;随着供俸植株发育阶段的提高诱导频率下降;最适基本培养基为 MS 培养基;蔗糖浓度从1.5%提高到9%,诱导频率逐渐下降;过高的维生素 B_1浓度对胚胎发生不利;2,4-D 的诱导效果优于 NAA,适宜的2,4-D 浓度为20ppm;光、暗处理与生长素种类和浓度之间存在交互作用;接种方式对诱导频率影响很大,体细胞胚只在下表皮与培养基接触的幼子叶的上表皮上产生,当上表皮与培养基接触时,两个表皮都不能产生体细胞胚;被试的所有基因型都能被诱导胚胎发生,不同基因型的诱导频率存在差异。     

大豆体细胞胚胎发生影响因素的研究

研究了影响大豆幼胚培养体细胞胚胎发生频率的9个因素。诱导体细胞胚胎发生的适宜幼胚长度为4mm;随着供体植株发育阶段的提高诱导频率下降;最适基本培养基为MS培养基; 蔗糖浓度从1.5％提高到9％,诱导频率逐渐下降;过高的维生素B1浓度对胚胎发生不利;2,4 — D的诱导效果优于NAA,适宜的2,4—D浓度为20ppm; 光、暗处理与生长素种类和浓度之间存在交互作用,接种方式对诱导频率影响很大,体细胞胚只在下表皮与培养基接触的幼子叶的上表皮上产生,当上表皮与培养基接触时,两个表皮都不能产生体细胞胚;被试的所有基因型都能被诱导胚胎发生,不同基因型的诱导频率存在差异。     

影响籼稻体细胞胚胎发生几个因素的研究

以 IR36、IR50、IR52及 IR54等品种的幼穗及成熟种子为材料,研究了蔗糖浓度、2,4-D、NAA、激动素及脱落酸对体细胞胚胎发生、结构的保持及植株分化的影响。6%蔗糖有利于胚性愈伤组织的诱导;3%的有利于胚性结构的保持及植株分化。当培养基中不含2,4-D,而含激动素与 NAA 时,幼穗直接出芽;当不含激动素而含2,4-D与 NAA 时,外植体产生非胚性愈伤组织;当不含 NAA 而含2,4-D 与激动素时,外植体产生胚性愈伤组织。认为,2,4-D与激动素是籼稻体细胞胚胎发生的基本因素,而 NAA 的作用是不明显的。不同外植体(幼穗与成熟种子)的体细胞胚胎发生,对2,4-D 与激动素的反应略有不同,幼穗更为敏感。在继代培养基中,加入低浓度的脱落酸有利于胚性结构的保持。随着继代世代的延续,分化培养中愈伤组织所表现出的绿色生长点状物不能发育成完整植株。     

向日葵的幼胚培养

实验所用的基本培基中以B_5培养基对向日葵幼胚培养为最适。激素萘乙酸主要促进幼胚的胚性生长。吲哚乙酸(IAA)与激动素(KT)的配合使用,对幼胚的分化起明显的调节作用。天然提取物则明显地促进幼胚生长、其中荸荠汁的培养基可使向日葵球形胚进行正常发育。 用两种改良的B_5培养基做胚胎培养系统,使向日葵种间杂交胚生长发育。先将杂交后的心形胚培养在胚胎生长培养基上,进行充分的胚性生长。然后把胚转移到萌发培养基,使其萌发形成幼苗,最后移入土壤直至开花。     

火炬松原生质体的体细胞胚胎发生

研究了基本培养基、原生质体密度和ABA浓度对火炬松（PinustaedaL．）悬浮细胞原生质体体细胞胚胎发生的影响。结果表明,DCR基本培养基最有利于原生质体的体细胞胚胎发生。体细胞胚胎发生所需的最适原生质体密度和ABA浓度分别是7×104个／mL和4mg／L。显微观察表明,来自原生质体的胚性胚柄细胞团（ESM：embryogenicsusPensormass）,经早期原胚（ESP：earlystageProembryos）阶段形成了后期原胚（LSP：latestageProembryos）。这一结果为火炬松的原生质体培养再生植株奠定了基础。     

蔗糖与激素对小麦幼穗体细胞无性系形成及生长特性的影响研究

通过在诱导培养基中添加不同浓度的蔗糖和激素,研究了蔗糖与激素对小麦幼穗体细胞无性系形成及生长特性的影响。结果表明;较高浓度的2,4-D有利于小麦幼穗愈伤组织的形成与生长,但低浓度的2,4-D更有利于胚性愈伤组织的形成,低浓度的KT能显著促进小麦幼穗愈伤组织的生长和胚性愈伤组织的形成,而高浓度的KT不利于小麦幼穗体细胞无性系的形成和生长;蔗糖有利于小麦幼穗胚性愈伤组织的形成,在2.5%-5.5%浓度范围内,随蔗糖浓度的提高,胚性愈伤组织的形成率也随之提高。     

白杄体细胞胚胎发生及其植株再生

白杄(Picea meyeri Rehd.et Wils.)的成熟胚在4～6℃低温下保存1个月后,接种于改良LP 2mg/L 2,4-D 1mg/L 6-BA的培养基上,黑暗条件下培养1个月便可产生白色半透明的胚性愈伤组织。整体染色封片观察表明,胚性愈伤组织由很多很长的胚柄细胞及其顶端的胚细胞团组成,这种愈伤组织培养物称为胚性胚柄团。胚性胚柄团在MS 1mg/L 2,4-D 1mg/L KT的继代培养基上黑暗条件下可保持旺盛的增殖能力和分化潜力。当胚性胚柄团转到MS 5mg/L ABA 5mg/L AgNO_3的分化培养基上,1个月后可产生大量正常的体细胞胚。体细胞胚成熟以后转到含0.5％活性炭的无激素1/2MS基本培养基上约40d后可长出1.5～2.5cm的根,约60d后可长出真叶。光、ABA、蔗糖及AgNO_3浓度是影响体细胞胚发生的主要因素。     

珍稀濒危植物香果树胚性细胞悬浮系的建立和植株再生

以香果树(Emmenopterys henryi)未成熟种子为试材, 探讨不同的接种量、基本培养基、糖浓度及植物生长调节物质等对体细胞胚胎生长的影响, 建立稳定的香果树胚性细胞悬浮培养与植株再生体系。结果表明: 悬浮培养条件下, 最适接种量为2%(鲜重); 较适合的基本培养基为MS; 蔗糖浓度为1%时容易使球形胚聚合体愈伤化, 浓度为3%和6%适合球形胚聚合体增殖, 浓度为9%则容易使球形胚聚合体褐化; 添加0.5 mg.L-16-BA 和0.5 mg.L-1 NAA的MS液体培养基, 当初始蔗糖浓度为3%, 然后逐步提高到6%则有利于香果树各个发育阶段的同步化; 子叶胚转到不含任何植物生长调节物质的MS固体培养基中可以长成正常植株。     

何首乌体细胞胚的诱导及植株再生研究

以何首乌茎尖、茎段为外植体,经体细胞胚发生途径,进行胚性愈伤组织诱导、体细胞胚的诱导、植株再生的研究.并采用临时压片法对体细胞胚的发育过程进行观察.结果表明愈伤组织诱导最适培养基为Ms＋6-BA 2.0 mg/L＋NAA 0.5 mg/L,体细胞胚诱导最适培养基为MS＋6-BA 1.0 mg/L＋NAA 0.2 mg/L.将产生的体细胞胚首先接种于MS基本培养基使其充分发育后转入MS＋6-BA 2.0 mg/L培养基中诱导出芽,出芽率高于直接采用Ms＋6-BA 2.0 mg/L培养基诱导.体细胞胚的发育过程是首先在愈伤组织表面形成许多瘤状突起即胚性细胞团,胚性细胞团继续发育成球形胚、盾形胚,球形胚、盾形胚成熟后发育成植株.     

培养条件对平贝母愈伤组织分化的影响

以平贝母无菌苗幼茎切段诱导产生的愈伤组织为试材,研究了基本培养基、蔗糖浓度、继代时间及继代周期时间对愈伤组织分化的影响。结果表明,ＭＳ基本培养基有利于不定芽的诱导,Ｎ６培养基有利于体细胞胚的诱导。在ＭＮ培养基上,随着蔗糖浓度的升高,不定芽发生能力下降,体细胞胚发生率升高。继代时间越长,不定芽及体细胞胚的发生能力均下降。体细胞胚胎发生的最佳继代周期时间是２１天,不定芽发生的最佳继代周期时间是２８天。     

Somatic embryogenesis from chickpea (Cicer arietinum L.) inmature cotyledons: The effect of zeatin, gibberellic acid and indole-3-butyric acid

Studies were conduced to test the effects of various cytokinins on somatic embryogenesis from chickpea (Cicer arietinum L.) immature cotyledons. Zeatin (13.7 μmol) added, to B5 basal medium, supplemented with 1.5 % sucrose and 0.2 μmol indole-3-acetic acid, was the most effective cytokinin. Lobular structures obtained from cotyledons cultures were transferred to B5 basal medium supplemented with gibberellic acid and indole-3-butyric acid at different concentrations. The most effective treatment was B5 medium containing 14.4 μmol gibberellic acid plus 1.0 μmol indole-3-butyric acid in which 42.8 % of lobular structures cultured formed normal somatic embryos. High conversion of embryos into plantlets (61.0–65.2 % embryos regenerated plants) was observed when germinated embryos were placed on plant development medium.     

影响大豆体细胞胚诱导因素的研究

体细胞胚的诱导是大豆体外再生的关键。基因型,诱导光周期,外植体的英位,蔗糖浓度等因素,可导致诱导频率及正常胚比例不同,影响植株再生。本研究选用黑龙江省主栽大豆基因型的未成熟子叶,在含高浓度生长素的MSB培养基上诱导体细胞胚产生。合丰25和东农7819为优选基因型,生育前期下部英位大小为2－4mm未成熟子叶体细胞胚发生效果最好;四种光周期下体细胞胚诱导频率相近,但连续弱光了正常胚比例高;NAA诱导优于2－4,D;10mg/1NAA与1．5％蔗糖配比组合最佳。     

Enhancement of somatic embryogenesis in camphor tree (Cinnamomum camphora L.): osmotic stress and other factors affecting somatic embryo formation on hormone-free medium

The aim of this study was to improve the direct somatic embryogenesis and initiate embryogenic callus formation in camphor tree (Cinnamomum camphora L.) on hormone-free medium. The influence of osmotic stress pretreatment of immature zygotic embryos (0.5 and 1.0 M solution of sucrose for 12, 24, 48, 72, 96, 120, and 144 h at 4 or 25°C) before cultured on hormone-free medium, on embryogenesis efficiency was assessed. The embryogenesis frequency was improved from 16.29 to 93.27%, while the average number of somatic embryos per explant increased from 3 to 12.57. Activated charcoal (AC), medium renewal, basal medium, light conditions and sucrose concentration in culture medium were also evaluated for their effect on somatic embryogenesis. AC addition and 10-day medium renewal did not increase embryogenesis efficiency significantly, and Murashige and Skoog (MS) medium proved to be more beneficial for somatic embryo formation than others. No differences were found between embryogenesis frequencies when cultured in darkness or under light, but culturing under light yielded more embryos. After the sucrose solution pretreatment, high level concentration of sucrose in induction medium was not needed for somatic embryogenesis, for it had a negative effect on somatic embryo formation when the concentration of sucrose was higher than 50 g l⁻¹. The derived embryogenic lines were maintained via repetitive embryogenesis on hormone-free medium. Low ratio formation of embryogenic callus was observed on the surface of somatic embryos both on induction and proliferation medium. Plantlets derived from somatic embryos grew vigorously with normal appearance similar to germinated zygotic embryos.     

青扦胚性细胞悬浮培养中影响体细胞胚发生因素的研究

试验以青扦（Ｐｉｃｅａｗｉｌｓｏｎｉ）的胚性愈伤组织为材料,以改良５９基本成分附加２４－Ｄ１ｍｇ／Ｌ及ＫＴ１ｍｇ／Ｌ为培养介质,比较了液体悬浮与半固体二种培养方式对胚性愈伤组织增殖和体细胞发生的影响,研究了液体悬浮培养过程中影响体细胞胚发生的因素。结果表明：液体悬浮培养好于半固体培养,它的胚性愈伤组织的生长率为２６８％,是半固体培养的１２４倍;体细胞胚的分化率为９３％,是半固体培养的２２倍;悬浮培养较佳的培养条件为：初始细胞密度为２％（鲜重）,蔗糖浓度为２０ｇ／Ｌ,摇床转速为１００ｒ／ｍｉｎ,ｐＨ为５８。经过两个月悬浮培养,将培养物转至１／２改良５９附加ＡＢＡ１ｍｇ／Ｌ的分化培养基上,３个月后每ｇ培养物上可获得２８５个正常的子叶期体细胞胚。     

Patterns of somatic embryo formation in Siberian larch: embryological aspects

Somatic embryogenesis was induced in Siberian larch by in vitro culturing zygotic embryos at different developmental stages. Cultures were grown in modified Murashige and Skoog medium supplemented with hormones 2,4-dichlorophenoxyacetic acid (2 mg/l) and 6-benzylaminopurine (0.5-1 mg/l). The success of somatic embryogenesis in this species depended on the tree genotype and developmental stage of embryos used for culturing. Somatic embryogenesis from immature zygotic embryos at the stage of cotyledon initiation was most active. After 5-10 days, such embryos formed the embryogenic tissue including two cell types--elongated highly vacuolated embryonic tubes and small embryonic cells. Somatic embryos were isolated from proliferating embryogenic tissues after 2 months of culture.     

Patterns of somatic embryo formation in Siberian larch: Embryological aspects

Somatic embryogenesis was induced in Siberian larch by in vitro culturing zygotic embryos at different developmental stages. Cultures were grown in modified Murashige and Skoog medium supplemented with hormones 2,4-dichlorophenoxyacetic acid (2 mg/l) and 6-benzylaminopurine (0.5–1 mg/l). The success of somatic embryogenesis in this species depended on the tree genotype and developmental stage of embryos used for culturing. Somatic embryogenesis from immature zygotic embryos at the stage of cotyledon initiation was most active. After 5–10 days, such embryos formed the embryogenic tissue including two cell types—elongated highly vacuolated embryonic tubes and small embryonic cells. Somatic embryos were isolated from proliferating embryogenic tissues after 2 months of culture.     

Regeneration of soybean (<Emphasis Type="Italic">Glycine max</Emphasis> L. Merrill) through direct somatic embryogenesis from the immature embryonic shoot tip

We describe here a simple and efficient system of soybean (Glycine max L. Merrill) regeneration through direct somatic embryogenesis by using immature embryonic shoot tips (IEST) as explants. The cultivar Kaohsiung 10 (cv. K10) used in this study did not show embryogenic response either from mature seed-derived explants (cotyledon, embryonic tip, leaf, shoot and root) or immature cotyledons. However, it showed a high percentage (55.8%) of somatic embryo (SEm) formation from the IEST excised 2–3 wk after flowering, thus indicating the crucial roles of type and age of explants. The IEST put forth primary SEm after 2 mo of culturing on Murashige and Skoog (MS) medium supplemented with 6% sucrose, 164.8 μM 2,4-dichlorophenoxyacetic acid (2,4-D), 5 mM asparagine and 684 μM glutamine. Subsequently, secondary SEm were developed 1 mo after culturing on MS medium containing 123.6 μM 2,4-D and 3% sucrose. Cotyledonary embryos were induced on MS medium supplemented with 0.5% activated charcoal after 1 mo. The embryos were desiccated for 72–96 h on sterile Petri dishes and regenerated on hormone-free MS medium. Plantlets with well-developed shoots and roots were obtained within 5–6 mo of culturing of IEST. The SEm-derived plants were morphologically normal and fertile. Various parameters thought to be responsible for efficient regeneration of soybean through somatic embryogenesis are discussed. To our knowledge, this is the first report to employ IEST as explants for successful direct somatic embryogenesis in soybean.     

华山松胚性愈伤组织诱导与幼胚离体培养

探索华山松(Pinus armandii)体细胞胚胎发生技术对其实施规模化无性繁殖和开展遗传转化具有重要意义。本文以1/2LM为基本培养基, 通过激素调节等措施对华山松的胚性愈伤组织诱导和幼胚的离体培养技术进行了初步研究。研究结果: 胚性愈伤组织诱导率最高可达52.71%, 但愈伤组织继代培养后没有体细胞胚胎的分化; 首次从其子叶期的幼胚中直接诱导出具有根和茎的完整植株, 诱导率达92%以上。文章确认了采集的幼胚发育状态对胚性愈伤组织的诱导有重要影响, 并对诱导的培养条件等进行了探讨。     

华山松胚性愈伤组织诱导与幼胚离体培养

探索华山松（Pinus armandii）体细胞胚胎发生技术对其实施规模化无性繁殖和开展遗传转化具有重要意义。本文以1／2LM为基本培养基,通过激素调节等措施对华山松的胚性愈伤组织诱导和幼胚的离体培养技术进行了初步研究。研究结果：胚性愈伤组织诱导率最高可达52．71％,但愈伤组织继代培养后没有体细胞胚胎的分化;首次从其子叶期的幼胚中直接诱导出具有根和茎的完整植株,诱导率达92％以上。文章确认了采集的幼胚发育状态对胚性愈伤组织的诱导有重要影响,并对诱导的培养条件等进行了探讨。