芍药花药愈伤组织诱导及体细胞胚发生

以芍药(Paeonia lactiflora)品种粉玉奴花药为外植体,研究不同浓度2,4-D对愈伤组织诱导、体胚发生及植株再生的影响,采用组织细胞学方法观察愈伤组织以及体细胞胚发育过程,采用根尖染色体法鉴定再生植株倍性。结果表明,芍药花药愈伤组织诱导的最适培养基为MS+1 mg·L–1 2,4-D+1 mg·L–1 N...     

大蕉(Musa paradisiaca ABB Linn.)花序愈伤组织的诱导及其体细胞胚发生

大蕉未成熟雄花接种到胚性愈伤组织诱导培养基中,4～5个月后可诱导出胚性愈伤组织,并可在继代培养基上增殖.胚性愈伤组织转移到体细胞胚诱导培养基中可诱导出体细胞胚.体细胞胚在成熟培养基上培养2个月后转移到含有0.2mg·L-1 6-BA的分化培养基上可以萌发,进而形成再生植株.组织学切片证明所诱导的愈伤组织是胚性组织,其所产生的体胚具有典型的单子叶植物体细胞胚的组织结构.     

芍药愈伤组织中体细胞胚发育过程的组织细胞学观察

以芍药（Paeonia lactiflora Pall.）3个芍药品种的茎段、叶片、叶柄为外植体,诱导体细胞胚发生,并采用石蜡切片法对该发育过程进行组织细胞学观察。结果表明：‘Going Bananas’的茎段愈伤诱导率达100%,增殖率在4.0以上,表现最佳;非胚性愈伤组织最佳诱导、增殖培养基为WPM+IAA 1.0mg·L^-1+6-BA 1.0mg·L^-1+NAA 0.5mg·L^-1+TDZ 0.5mg·L^-1+CH 0.625g·L^-1;愈伤组织转入到1/2 MS（Ca²⁺加倍）+2,4-D 2.0mg·L^-1+ABA 0.5mg·L^-1或ZT 1.0mg·L^-1的培养基中,连续暗培养90后得到胚性愈伤;之后转入到成熟培养基1/2MS（Ca²⁺加倍）+6-BA 1.0mg·L^-1+NAA 0.2mg·L^-1中,见光培养60d,逐渐发育至球形胚和心形胚。在石蜡切片观察中,芍药的体细胞胚起源方式包括外起源和内起源两个途径。在外起源方式中包括单个表层细胞的外起源和多个表层下细胞的共同起源。3种方式的区别主要在于起始的位置和起始细胞数量,后期均形成原胚结构。胚性细胞的分裂方式为对称分裂,未发现不对称分裂细胞的存在。     

亚麻品种'双亚5号'的胚性愈伤组织诱导和体细胞胚胎发生

亚麻品种'双亚5号'的种子在MS 1.0 mg·L-1 2,4-D 30 g·L-1蔗糖培养基上可诱导出愈伤组织,将其转入MS 0.5mg·L-1KT 0.5mg·L-1NAA培养基上培养10周后诱导出大量胚性愈伤组织,结构较致密,浅黄色,表面有成团的紧密粘附在一起的小颗粒形态.但其继代周期不宜太长,继代次数不宜多,否则易回到非胚性化状态.胚性愈伤组织转A.MS 1.5 mg·L-1(T 1.0 mg·L-1 2,4-D分化培养基上培养40 d后可获得大量的球形胚状体.少量球形体胚可萌发形成正常的子叶胚初期形态,较多的球形体胚形成次生体胚或仅有单极性的畸形胚状体.组织解剖学观察表明,诱导出的是亚麻胚性愈伤组织和胚状体.     

Somatic Embryogenesis and Plant Regeneration from Leaf Callus of Hordeum vulgare

Explants obtained from the basal portion of leaves of Hordeumvulgare (cv. Karan 92) gave rise to callus when cultured onMurashige and Skoog (MS) basal medium supplemented with 2, 4-dichlorophenoxyaceticacid (2, 4-D). Initially, the callus was friable, shiny-whiteand watery but subsequently some compact, nodular callus appeared.The latter were cultured on MS medium containing 0.05 mg l^–12, 4-D and 0.1 mg l^–1 N6-furfurylaminopurine (kinetin),when plantlets were generated. Histological studies showed thatplantlet regeneration occurred by the formation of somatic embryos.The regenerated plants had the normal diploid chromosome number(2n = 14). Hordeum vulgare, barley, somatic embryogenesis, tissue culture, plant regeneration     

Plant Regeneration Through Somatic Embryogenesis in Leaf Derived Callus of Plumbago Rosea

Regeneration of Plumbago rosea L., a rare medicinal plant, via somatic embryogenesis in callus cultures derived from leaf explants was described. Optimum callus formation was achieved on semi-solid Murashige and Skoog (MS) medium supplemented with 0.25 mg dm^–3 kinetin and 2.0 mg dm^–3 1-naphthaleneacetic acid (NAA). Somatic embryogenesis was achieved upon transferring the 4-week-old callus to a medium containing 1.0 mg dm^–3 kinetic (Kn), 0.5 mg dm^–3 gibberellic acid (GA₃) and 0.1 mg dm^–3 NAA. Embryo maturation and germination was achieved on the half-strength MS basal salts supplemented with 0.01 – 0.25 mg dm^–3 Kn and 2 % (m/v) saccharose. An average of 50 – 60 plantlets were obtained from 150 mg of embryogenic callus within 4 week of subculture. Out of the 50 plantlets about 28 survived in the greenhouse.     

Label-Free Quantitative Proteomics of Embryogenic and Non-Embryogenic Callus during Sugarcane Somatic Embryogenesis

The development of somatic cells in to embryogenic cells occurs in several stages and ends in somatic embryo formation, though most of these biochemical and molecular changes have yet to be elucidated. Somatic embryogenesis coupled with genetic transformation could be a biotechnological tool to improve potential crop yields potential in sugarcane cultivars. The objective of this study was to observe somatic embryo development and to identify differentially expressed proteins in embryogenic (E) and non-embryogenic (NE) callus during maturation treatment. E and NE callus were cultured on maturation culture medium supplemented with different concentrations (0.0, 0.75, 1.5 and 2.0 g L^-1) of activated charcoal (AC). Somatic embryo formation and differential protein expression were evaluated at days 0 and 21 using shotgun proteomic analyses. Treatment with 1.5 g L^-1 AC resulted in higher somatic embryo maturation rates (158 somatic embryos in 14 days) in E callus but has no effect in NE callus. A total of 752 co-expressed proteins were identified through the SUCEST (The Sugarcane EST Project), including many housekeeping proteins. E callus showed 65 exclusive proteins on day 0, including dehydrogenase, desiccation-related protein, callose synthase 1 and nitric oxide synthase. After 21 days on maturation treatment, 14 exclusive proteins were identified in E callus, including catalase and secreted protein. NE callus showed 23 exclusive proteins on day 0 and 10 exclusive proteins after 21 days on maturation treatment, including many proteins related to protein degradation. The induction of maturation leads to somatic embryo development, which likely depends on the expression of specific proteins throughout the process, as seen in E callus under maturation treatment. On the other hand, some exclusive proteins can also specifically prevent of somatic embryos development, as seen in the NE callus.     

Requirement of Ethylene for Growth of Callus and Somatic Embryogenesis in Medicago sativa L.

The role of ethylene in the growth of callus and somatic embryogenesisin Medicago sativa was examined. The application of 2,5-norbornadiene,a competitive inhibitor of ethylene action, during a 10 d inductionperiod to medium containing 2,4-D and kinetin inhibited thegrowth of callus but did not affect somatic embryogenesis, nordid it affect ethylene production during the induction stage.The exposure of tissue, incubated on differentiation medium,without hormones, to an atmosphere of 2,5-norbornadiene, inhibitedboth growth and embryo maturation and stimulated pigmentation.The inhibition of embryo maturation was observed even in thepresence of norbornadiene at a concentration which did not affectgrowth of tissue. It is suggested that the action of endogenous ethylene is necessaryfor the growth of the callus and embryo maturation. Key words: Medicago sativa, ethylene, callus growth, somatic embryogenesis     

逆境处理和DNA甲基化影响柑橘体细胞胚发生

对 1 5种柑橘胚性愈伤组织进行体细胞胚诱导 ,发现逆境处理有利于体细胞胚发生 ,并可以恢复部分品种的体细胞胚发生能力。对具有和失去体细胞胚发生能力的两种纽荷尔脐橙 (CitrussinensisOsb .)愈伤组织进行随机扩增多态性DNA (RAPD)分析没有检测到带型的差异 ,而对它们的甲基化敏感扩增多态性 (MSAP)进行分析则发现两种愈伤组织间具有明显的DNA甲基化差异 ,具体细胞胚发生能力的愈伤组织的甲基化水平较失去体细胞胚发生能力的低     

逆境处理和DNA甲基化影响柑橘体细胞胚发生

对15种柑橘胚性愈伤组织进行体细胞胚诱导,发现逆境处理有利于体细胞胚发生,并可以恢复部分品种的体细胞胚发生能力.对具有和失去体细胞胚发生能力的两种纽荷尔脐橙( Citrus sinensis Osb.)愈伤组织进行随机扩增多态性DNA (RAPD) 分析没有检测到带型的差异,而对它们的甲基化敏感扩增多态性 (MSAP) 进行分析则发现两种愈伤组织间具有明显的DNA甲基化差异,具体细胞胚发生能力的愈伤组织的甲基化水平较失去体细胞胚发生能力的低.     

暴马丁香成熟胚愈伤组织和体胚诱导及其生理状态解析

以暴马丁香成熟胚为外植体进行愈伤组织和体胚发生诱导,通过调节诱导培养基中植物生长调节剂的种类和浓度,分析其对愈伤组织诱导和体胚诱导的影响,同时对培养过程中的外植体进行形态发生观察和生理状态分析。结果表明:①暴马丁香成熟胚外植体培养30 d可见直接体胚发生、60 d可见子叶型体胚;②BA在愈伤组织诱导过程中起到了主导作用,在0.5 mg·mL^-1BA和5~6 mg·mL^-1NAA组合下,愈伤组织诱导率可达100%;在0.5 mg·mL^-1BA和5 mg·mL^-1NAA组合体胚诱导率可达8%;③多酚含量在愈伤组织形成初期急剧上升且在培养过程中保持较高水平,子叶型胚期PAL和POD活性升高、MDA和SOD活性略下降。     

Effect of MGBG on Growth of the Callus Tissue,Induction of Somatic Embryogenesis of Alfalfa and Their Ethylene Biosynthesis

MGBG [methylglyoxal bis(guanylhydrazone)], when added to the initial of callus subculture, promoted ethylene production in callus grawn in Bsh medium, and enhanced the ACC (1-aminocyclopropane-l-carboxylic acid) level and the ACC synthase activity both in the callus grown in Bsh medimn, and in the suspension cultures in Bsg liquid medium for induction of somatic embryogenesis. However, MGBG reduced the malonyl-ACC (MACC) level of the suspension tissues. The treatments of MGBG caused growth of the cultured tissues and induction efficiency of somatic embryogenesis to reduce. Moreover, the effects of MGBG was enhanced when it was added at induction medium of somatic embryogenesis. It could be concluded that this inhibitory effect of MGBG could be due to its promotion of ethylene, ACC levels and ACC synthase activity which has shown to inhibit somatic embryogenesis. The reduction of MACC level could be also involved in the MGBG effect on induction of the embryogenesis.     

Callus Culture of Sainfoin (Onobrychis viciifolia) and Plant Regeneration through Somatic Embryogenesis

Callus of sainfoin (Onobrychis viciifolia Scop.) was initiatedfrom stem and root explants which were obtained from seedlingsgrowing in vitro, on Linsmaier Skoog (LS) medium supplementedwith 1 mg l^–1 2, 4-D and 1 mg l^–1 BA or only 1 mgl^–1 BA, and the Vacin and Went medium without hormones.Somatic embryos were formed on LS medium containing 1 m l^–1BA. Embryos developed into complete plants on filter paper saturatedwith hormone-free LS medium. Onobrychis viciifolia, somatic embryogenesis, callus culture, plant regeneration     

Somatic Embryogenesis and Plantlet Regeneration from Callus of Mature Zygotic Embryos of Masson Pine

Embryogenic cultures were initiated from mature zygotic embryos of masson pine (Pinus massoniana Lamb. ) on DCR medium supplemented with 2, 4-D 10 mg/L, KT and BA each at 4 mg/L. Pale and translucent calli with early stage proembryos were maintained and multiplicated on DCR medium supplemented with 2, 4-D 1.0 mg/L KT and BA each at 0.4 mg/L. Robust late-stage proembryos were obtained when the calli were cultured on DCR medium containing 9000 mg/L myo-inositol. Abscisic acid and activated charcoal promoted the formation of cotyledonary embryos at the highest frequency of 35.1 %. Mature somatic embryos could germinate and develop further into plantlets when they were isolated and cultured on a hormone-free DCR medium.     

胡桃楸胚性愈伤组织诱导与体细胞胚胎发生

胡桃楸是东北东部山地阔叶红松林的重要组成树种。因其被大量采伐,资源日趋枯竭。体细胞胚胎发生是快速繁殖和人工种子研制的基础,对遗传改良有重要意义。为探讨不同外植体、植物生长调节物质种类及配比对胡桃楸培养物的影响,建立了胡桃楸体胚发生及再生植株体系。结果表明:合子胚为外植体时最易形成胚性愈伤组织,外植体最佳取材时期为5～6月。胡桃楸胚性愈伤组织最适诱导为MS+1.0mg·mL-12,4-D+0.5mg·mL-16-BA;体细胞胚的诱导、发育和分化的适宜的培养基为附加蔗糖60g.L-1、水解酪蛋白700mg·mL-1时不添加任何生长调节物质的MS培养基。     

落叶松体细胞胚胎发生诱导多倍体的研究

通过检测细胞系增值率及细胞分裂指数确定细胞系分裂最旺盛时期,利用秋水仙素和氨磺灵诱导落叶松多倍体体细胞胚胎的发生,揭示了两种抗微管物质对落叶松体细胞胚胎发生的影响.秋水仙素在浓度为500 mg·L-1,浸泡处理36 h时能使多倍体体细胞胚胎发生的比例达到85.2%,体细胞胚发生率达到351.1个/g;氨磺灵在1～5 mg·L-1的浓度下处理后的细胞系几乎失去体细胞胚分化能力,无法大量获得多倍体体细胞胚.通过细胞压片及去壁-低渗涂片对比观察发现氨磺灵诱导后的细胞系除了染色体数目加倍外还有部分细胞发生程序性死亡.该研究初步证明了秋水仙素较适合落叶松多倍体体细胞胚的诱导,氨磺灵对落叶松毒害明显,不适合其多倍体的诱导.