地锦体细胞胚胎发生研究

以地锦幼胚为外植体,接种在附加6 BA2mg·L-1及2,4 D0～4mg·L-1的改良B5培养基中诱导愈伤组织,愈伤诱发率0～100% 对诱导出的愈伤组织采用6 BA和2,4 D不同浓度组合的培养基进行继代,对能稳定继代的愈伤组织,以改良B5为基本培养基进行分化培养,采用L16(44)正交设计,研究6 BA、NAA、AC及LH四因素不同浓度组合对体细胞胚胎发生的影响。结果表明:适宜地锦幼胚体细胞胚胎发生的培养基为:愈伤组织诱导:改良B5 6 BA2 0mg·L-1 2,4 D2 0mg·L-1;愈伤组织继代:改良B5 6 BA2 0mg·L-1 2,4 D1 0mg·L-1;胚状体分化及萌发:改良B5基本培养基。胚状体诱发率33%,成苗率53%,移栽成活率85%。胚状体为外起源发生方式,经球形胚、心形胚、鱼雷形胚、子叶形胚等阶段发育成熟。     

重瓣非洲菊的组培技术研究

取重瓣非洲菊幼小花托 ,以MS为基本培养基 ,附加不同生长调节剂及浓度水平诱导愈伤组织形成不定芽进行组培研究。结果表明 :重瓣非洲菊幼小花托在MS +BA 5 0mg·L-1+NAA 0 5mg·L-1培养基上能很好地诱导形成愈伤组织并分化出不定芽 ,诱导率 91 8% ;MS +KT 5 0mg·L-1+IAA 0 5mg·L-1培养基对不定芽增殖效果较好 ,增殖倍数达4 4 4 ;生根培养基为 1/ 2MS +NAA 1 0mg·L-1,生根率达 98%。选择河泥 +细沙 (2∶1)混合作移栽基质 ,成活率达 90 %以上。     

日本落叶松体细胞胚胎发生的研究

日本落叶松日永 10、日永 6 9和日草 34三个优株 ,将其未成熟种子和成熟种子的胚及成熟种子萌发的胚根、胚轴和绿色愈伤组织 5种外植体进行体细胞胚胎发生的试验 ,其中只有未成熟胚诱导出胚性愈伤组织 ,筛选出1)胚性愈伤组织诱导培养基 :S培养基 6_BA 0 5mg·L- 1 KT 0 5mg·L- 1 2 ,4_D 1 0mg·L- 1 ;2 )胚性愈伤组织保持与增殖培养基 :S培养基 6_BA 0 2 5mg·L- 1 KT 0 2 5mg·L- 1 2 ,4_D 0 1mg·L- 1 ;3)成熟体细胞胚诱导培养基 :S培养基 ABA 15mg·L- 1 PEG4 0 0 0 10 0g·L- 1 。以上 3种培养基均需附加蔗糖 30g·L- 1 ,谷氨酰胺 4 5 0mg·L- 1 ,水解酪蛋白 5 0 0mg·L- 1 ,凝胶 (Sigma#) 3g·L- 1 ,pH5 8。 4 )体细胞胚萌发培养基 :S培养基 ,不加任何激素 ,附加蔗糖 2 5g·L- 1 ,凝胶 (Sigma#) 3g·L- 1 ,pH5 8。     

大果榉未成熟胚愈伤组织的诱导和植株再生

将大果榉的未成熟胚培养在不同植物激素种类和浓度的术本植物培养基(WPM)上产生愈伤组织,然后将愈伤组织接种在芽的诱导培养基上形成不定芽,不定芽在生根培养基上生根形成完整植株．结果表明:大果榉愈伤组织诱导的适合培养基为WPM附加BA 1．0 mg·L-1+NAA 1．0 mg·L-1,诱导事为59．1％;不定芽的分化培养基为在WPM+NAA 0．5 mg·L-1+6．BA 2．0 mg·L-1,不定芽的诱导率为71．1％,在WPM+BA2．5 mg·L-1+NAA1．0 mg·L-1的培养基种芽分化的平均数未为5．21±1．02;生根培养基以WPM+0．1 mg·L-1 IBA的效果量好,生根率可达63．33％,平均最多生根条数为．再生植株在温室适应培养的存活率为92％．     

油茶离体培养诱导再生植株的研究

采用普通油茶Camelliaoleifera优良无性系湘林4号为实验材料,研究了离体条件下带芽的茎段、幼胚、子叶的愈伤组织的形成和植株再生技术。茎段离体培养以MS为基本培养基,附加6-BA3.0mg·L-1+NAA1.0mg·L-1对芽进行诱导,诱导率达到83.33%;继续在此培养基上继代得到丛生芽,增殖比例为1∶20。对于幼胚和子叶,附加2,4-D2.0mg·L-1+KT1.0mg·L-1诱导愈伤组织得到很好的效果,将愈伤组织转到附加6-BA3.0mg·L-1+NAA0.05mg·L-1和6-BA2.5mg·L-1+IAA1.5mg·L-1的培养基上诱导出不定芽,诱导率达到90%以上。芽苗增殖以MS+6-BA2.5mg·L-1+IAA1.5mg·L-1的培养基为好。将无菌苗在MS培养基(附加NAA7mg·L-1)上诱导生根,生根率达93%。     

刺槐未成熟合子胚的体细胞胚胎发生和植株再生

以刺槐不同胚龄的未成熟合子胚为外植体,采用混合正交试验设计,研究幼胚胚龄和不同外源激素种类及质量浓度对刺槐胚性愈伤组织的诱导和体细胞胚分化、萌发的影响.结果表明:开花后8周(55天左右)是胚性愈伤组织和体胚诱导的最佳外植体取材时期;MS+2,4-D 5.0 mg·L-1 +BA0.5 mg·L-1是诱导胚性愈伤组织的最佳培养基,出愈率最高为95.42％±0.02％;MS +NAA0.5 mg·L-1 +BA0.5 mg·L-1+谷氨酰胺250 mg·L-1+水解酪蛋白500 mg·L-1是体细胞胚诱导和分化的最佳培养基,直接体细胞胚发生率最高为92.40％±0.12％,通过愈伤组织诱导体细胞胚发生的频率最高为90.73％ ±0.49％.一旦形成球形胚,将培养物转接到不含任何生长调节剂的MS培养基上,体细胞胚经成熟萌发可进一步形成完整小植株.     

斑马水塔花的组织培养和快速繁殖

试验筛选出斑马水塔花的诱导愈伤组织培养基MS+6-BA1.0mg·L-1+NAA0.2mg·L-1，愈伤组织诱导率达96.7%；继代增殖培养基MS+6-BA2.0mg·L-1+NAA0.2mg·L-1，增殖倍数达8～9倍；生根培养基MS+KT 0.5mg·L-1+NAA0.5mg·L-1，生根率达100%。幼苗经炼苗后，移栽到经菌肥处理的甘蔗渣∶泥炭土∶珍珠岩＝6∶3∶1的基质上，成活率达95%以上。     

乐东拟单性木兰茎段愈伤组织诱导与褐变控制的研究

利用乐东拟单性木兰成龄树嫩枝茎段作外植体,研究了控制褐变和组织培养的技术。试验结果表明,抗氧化剂控制褐变的效果比吸附剂好,以抗坏血酸最好,其次是巯乙醇和二硫苏糖醇。在培养基中分别加入抗坏血酸(10mgL-1)、巯茎乙醇(1mL·L-1)和二硫苏糖醇(1 5mg·L-1)使外植体的褐变率分别比对照降低67%、53%和33%。在接种前,将外植体在4℃下冷处理2h,能使外植体的褐变率从对照的35%降低为25%。1 2MS和WPM均是较合适的基本培养基。外植体在BA1 5mg·L-1+NAA0 2mg·L-1+2,4 D1 0mg·L-1的激素组合下,能诱导出较多的愈伤组织;在WPM+BA1 5mg·L-1+KT0 5mg·L-1+NAA0 2mg·L-1+2,4 D0 2mg·L-1的培养基上,能产生较多的不定芽和愈伤组织;WPM+BA1 5mg·L-1+KT0 5mg·L-1+NAA0 2mg·L-1+IAA1mg·L-1的培养基适合于芽增殖;在1 2MS+NAA0 3mg·L-1+IBA0 2mg·L-1的培养基上,添加15g·L-1活性炭有利于生根。     

红阳猕猴桃茎段愈伤组织诱导成苗技术

以红阳猕猴桃为试材 ,通过田间植株茎段愈伤组织诱导、分化培养 ,研究其成苗技术 ,试验发现 ,MS +BA 1 0mg·L-1+NAA 0 1mg·L-1可以获得 88%的愈伤组织诱导率 ,而且所得到的愈伤组织易于分化成苗。在MS +BA 2 0mg·L-1+NAA 0 1mg·L-1中 ,芽分化率达到 5 6 6 %。 1/ 2MS +NAA 0 2mg·L-1为生根培养基     

橙色红千层愈伤组织诱导技术的研究

采用正交设计的方法研究了植物生长调节剂配比、蔗糖质量浓度等对橙色红千层组织培养中愈伤组织形成的影响。结果表明:MS+6 BA5 0mg·L-1+KT4 0mg·L-1+IBA0 5mg·L-1+蔗糖30g·L-1是最适培养基。     

Somatic Embryogenesis and Plantlet Regeneration in Callus Cultures Derived from Mature Zygotic Embryos of Slash Pine

1TheworkwassupportedbyChineseNationalHighTechnologyDevelopmentProgram"863project"IntroductionSomaticembryogenesishasagreatpotentiaIforrapidProPaationofsuPCriorconifcrspccics.ThefirstreportonsomaticembryogenesisandplantlctrcgencrationinconifersPeCieswasach…     

Somatic embryo production and plant regeneration of Japanese black pine (Pinus thunbergii)

Somatic embryogenesis in Pinus thunbergii was initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of somatic embryos were obtained on maturation media containing maltose, activated charcoal, abscisic acid, and polyethylene glycol as osmotic agent. The best result among the cell lines tested was achieved with the cell line T-205-3. More than 900 somatic embryos per petri dish, on average, were obtained after about 8 weeks of culture on maturation medium. Sixty percent of somatic embryos tested germinated after transfer to plant growth regulator-free medium and then 85% of them converted into plantlets.     

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.     

In vitro somatic embryogenesis in callus cultures of Azadirachta indica A. Juss.—a multipurpose tree

Plant regeneration via somatic embryogenesis was achieved in embryogenic callus cultures derived from immature zygotic embryos 40 days after anthesis of Azadirachta indica A. Juss. on semisolid basal Murashige and Skoog (MS) salts and vitamins supplemented with 1.11 µM 6-benzylaminopurine (BA) and 4.52–6.78 µM 2,4-dichlorophenoxyacetic acid (2,4-D). The globular-stage embryos were induced when the callus was transferred to medium with 1.11 µM BA and 0.45 µM 2,4-D. The highest average number of somatic embryos per 200 mg of callus was 152.8 after 8 weeks of culture on the medium. Maturation and germination of the somatic embryos were achieved on half-strength MS salts and vitamins supplemented 0.38–0.94 µM abscisic acid (ABA) and 2% (w/v) sucrose. The maximum percentage (64.2%) of germination was obtained with 0.94 µM ABA within 2 weeks of culture. Somatic embryo-derived plantlets were acclimatized in a greenhouse and subsequently showed normal growth. This efficient protocol will be helpful for propagating elite clones on a mass scale and will also be useful for genetic transformation study.     

火炬松胚性愈伤组织诱导和植株再生的研究

火炬松胚性愈伤组织诱导和植株再生的研究唐巍欧阳藩（中国科学院化工冶金研究所生化工程国家重点实验室北京１０００８０）郭仲琛（中国科学院植物研究所北京１０００９３关键词火炬松,体细胞胚胎发生,植株再生本文于１９９６年１０月２８日收到。国家“８６３”资...     

Efficient plant regeneration of Hinoki cypress (Chamaecyparis obtusa) via somatic embryogenesis

This report describes the efficient plant regeneration of Chamaecyparis obtusa Sieb et Zucc. via somatic embryogenesis. Embryogenic cultures were initiated from megagametophytes containing immature zygotic embryos. Embryogenic cultures were maintained and proliferated by 2–3-week interval subcultures in medium supplemented with 2,4-dichlorophenoxyacetic acid and 6-benzylaminopurine. High maturation frequencies of cotyledonary embryos were obtained on maturation medium containing maltose, polyethylene glycol, activated charcoal, and abscisic acid. Somatic embryos germinated readily after transfer to plant growth regulator-free medium. Growth of regenerated emblings has been monitored in a greenhouse.     

Organogenesis and embryogenesis in mature zygotic embryos of Picea sitchensis

Differentiation of adventitious buds and somatic embryos from mature zygotic embryos of Picea sitchensis (Bong.) Carr. is described. Adventitious buds were formed on embryos pulse-treated with 250 microM benzyladenine for 2 h and cultured on medium lacking growth regulators. Buds were initiated at different frequencies and sites depending on when the BA-pulsed embryos were transferred to fresh culture medium. Embryogenic callus was formed when the zygotic embryos were cultured on medium containing 10 microM 2,4-dichlorophenoxyacetic acid and 5 microM benzyladenine. Although 50% of the embryos gave rise to embryogenic callus, only 20% of the callus continued to proliferate after subculture. Proliferation of new somatic embryos occurred from both the embryonic region and the suspensor region on previously formed somatic embryos. The pattern for development of adventitious buds and somatic embryos in Picea sitchensis is compared to that in Picea abies under similar culture conditions.     

火炬松体细胞胚胎发生和干化体细胞的氧化物酶活性(英文)

培养于附加2,4-D、BA和KT的愈伤组织诱导培养基上的火炬松成熟合子胚在培养3-9周后形成白色、半透明、有光泽的粘性愈伤组织。这类愈伤组织形成于成熟合子胚的子叶,但当用NAA或者IBA代替愈伤组织诱导培养基中的2,4-D时,它的诱导频率明显降低。这种粘性愈伤组织在分化培养基上形成体细胞胚。体细胞胚经过去50μm ABA和8.5%PEG600处理后成为耐干化胚。扫描电镜观察表明,萌发处理36小时后,耐干化胚恢复到干化处理之前的状态且大小和形态正常,而不耐干化胚不能恢复到干化处理之前的状态且表面撕裂。过氧化物酶活性的分析结果表明,耐干化胚有更高的过氧化物酶活性。耐干化胚的高过氧化物酶活性可能与催化H2O2的分解和保护体细胞胚免受氧化的伤害有关。     

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).