长江下游甘蓝型油菜小孢子的培养体系

自从德国学者首次报道甘蓝型油菜小孢子培养技术成功以来[1],该项技术已被迅速采用.小孢子培养能够得到基因型纯合的甘蓝型油菜株系(双单倍体,Double haploid),利用该技术可以迅速纯合保持系和恢复系[2],避免性状分离;也可以对远缘杂交后代通过加倍来稳定外缘染色体[3];还可以结合诱变技术进行油菜种质资源创新,目前已经获得品质和抗性性状非常优良的株系[4,5].     

Tissue Culture Response from Seedling Explants of Commercial Barley Cultivars Grown in Bulgaria

Abstract

Only a few studies have been conducted in which regenerability of barley has been examined. In the current study, 17 barley genotypes (nine two-row malting type: Aster, Emon, Ruen, Jubiley, PV101, Körten, Krassi 2, Perun and Igri, and eight six-row feed types: Karnobat, Hemus, Jerun, Veslets, Aheloi 2, Diana, Panagon, and Izrgev) were evaluated for tissue-culture response from seedlings during a three-year period. Regenerable calli were obtained from all tested genotypes. Although there was much variation in regeneration capacity among the cultivars, plants were obtained from all cultivars. The majority of green plants grown to maturity were fertile and normal in appearance. The frequency of embryogénie callus induction and regenerability was influenced by genotype and growth conditions of the donor plants. Genotype was the most important determinant of the in vitro response. The best in vitro performance, based on ability to form regenerable calli and to regenerate plants was observed for Ruen, Aster, and Emon.     

不结球白菜游离小孢子培养及再生植株的倍性鉴定

对影响不结球白菜小孢子培养和植株再生的几个因素及再生植株的倍性进行分析.结果表明:基因型是影响小孢子胚诱导率的最重要因子,正反交材料的胚诱导率无显著差异;供体植株开花后6～10 d培养可获得高的胚诱导率;琼脂在8～12 g·L-1范围内,随着质量浓度的提高可显著提高小孢子胚再生率.结合流式细胞仪和植株形态鉴定,不结球白菜小孢子再生植株自然加倍率总体水平较高,有的高达90%,但不同基因型间有明显差异.     

光敏核不育水稻农垦58S细胞学研究

光敏核不育水稻农垦58S小孢子败育发生在单核晚期，败育在很短时间内完成。败育前，小孢子大量液泡化，最后只乘空腔。败育小孢子不能正常形成花粉外壁。花药绒毡层的异常，体现在解体延迟，内含物丰富，富含嗜锇颗粒，乌氏体及孢粉素不参与花粉壁的形成，绒毡层异常是农垦58S败育的重要原因。     

华山矾大小孢子的发育和雌雄配子体的形成

研究了华山矾大、小孢子发育和花粉壁的形成以及大孢子的发生和雌配子体的发育过程。华山矾的花药四室。绒毡层腺质型 ,发育后期出现双核至多核。小孢子四分体四面体型 ,胞质分裂连续型。成熟花粉具 2细胞。子房 2室 ,中轴胎座。胚珠倒生 ,单珠被 ,薄珠心 ,胚囊发育为蓼型。该研究为山矾科的胚胎学提供了新资料。     

木薯体细胞胚发生过程中抗氧化酶活性变化

以木薯成熟体胚的子叶胚为外植物诱导的体细胞胚为材料,研究其体细胞胚发生过程中抗氧化酶活性(SOD,POD,CAT活性)和过氧化物酶同工酶酶谱的变化。结果表明,木薯体细胞胚发生过程中,SOD活性均在胚性细胞向球形胚转化时下降,鱼雷形胚和子叶形胚发育时升高;而POD和CAT活性呈相反趋势。初步认为,SOD酶活性降低可作为木薯胚性细胞初期分化和胚胎早期发育的一个判断指标。在木薯胚性细胞发生到分化和成熟过程中,过氧化物酶同工酶谱带由5条上升为9条,其中有5种过氧化物酶同工酶稳定出现,这可能是维持细胞基础代谢的一些基因表达的产物。     

Functional genomics of microspore embryogenesis

Isolated plant microspores, when stressed and cultured in vitro, can be diverted from their normal gametophytic pathway towards sporophytic development, with the formation of haploid embryos and ultimately doubled-haploid plants. This process is called androgenesis or microspore embryogenesis, and is widely used in plant breeding programmes to generate homozygous lines for breeding purposes. Protocols for the induction of microspore embryogenesis and the subsequent regeneration of doubled haploid (DH) plants have been successfully developed for more than 200 species. These practical advances stand in stark contrast to our knowledge of the underlying molecular genetic mechanism controlling this process. The majority of information regarding the genetic and molecular control of the developmental switch from gametophytic to sporophytic development has been garnered from four intensely studied (crop) plants comprising two dicotyledonous species, rapeseed (Brassica napus) and tobacco (Nicotiana tabacum), and two monocotyledonous species, wheat (Triticum aestivum) and barley (Hordeum vulgare). In these species the efficiency of microspore embryogenesis is very high and reproducible, making them suitable models for molecular studies. In the past, molecular studies on microspore embryogenesis have focussed mainly on the identification of genes that are differentially expressed during this developmental transition and/or early in embryo development, and have identified a number of genes whose expression marks or predicts the developmental fate of stressed microspores. More recently, functional genomics approaches have been used to obtain a broad overview of the molecular processes that take place during the establishment of microspore embryogenesis. In this review we summarise accumulated molecular data obtained in rapeseed, tobacco, wheat and barley on embryogenic induction of microspores and define common aspects involved in the androgenic switch.     

不结球白菜小孢子胚胎发生过程及发育途径研究

采用FDA荧光染色法对小孢子游离培养的发育过程进行了观察。结果表明:不结球白菜花粉母细胞减数分裂释放出的小孢子发育到单核靠边期及稍后时期染色最深、活力最强。多数游离小孢子培养经历的发育过程与合子胚相似,并且胚胎发育迅速,6～9 d产生大量球型胚,13 d开始出现子叶型胚胎。小孢子胚胎发育途径既有对称分裂并产生胚胎的途径(B途径),也有第1次不对称有丝分裂的方式;第1次不对称分裂后,既有营养细胞单独发育成胚的现象(A途径),也有营养细胞和生殖细胞共同发育成胚的现象(E途径),但未发现生殖细胞单独发育成胚的现象。     

棉花优良品种中棉所27直接胚胎发生与植株再生

选用我国难培养基因型优良棉花品种中棉所27进行组织培养,研究了该品种体细胞胚胎发生与植株再生能力。该品种体细胞胚胎发生有2种情况:一是由外植体直接诱导获得胚性愈伤组织并形成不同发育时期的胚状体;二是先诱导获得非胚性愈伤组织,再经多次继代培养,获得胚胎发生。不同激素及其组合和浓度配比是影响中棉所27直接诱导获得胚性愈伤组织及体细胞胚胎发生的直接因素。另外,饥饿培养及继代周期对胚胎发生也有重要影响。0.1mg/LKT和0.1mg/LIAA组合、0.1mg/LKT和0.1mg/LZT组合、0.1mg/LZT单独使用,在饥饿条件下培养,均能直接高效诱导获得大量胚性愈伤组织,并有短期内形成不同发育时期的胚状体。胚状体在不含任何激素的MS B5培养基上培养有利于植株再生,并降低畸形苗形成率。另外,还发现不同培养基对中棉所27胚性愈伤组织增殖、体细胞胚形成和萌发的影响不同。     

转MsLEA4-4基因拟南芥株系的耐盐性分析

为了深入研究紫花苜蓿（Medicago sativa）MsLEA4-4基因的抗逆性,本试验对前期获得的转MsLEA4-4基因的拟南芥（Arabidopsis thaliana）株系进行不同浓度盐胁迫处理,以验证MsLEA4-4基因在拟南芥植株体内的耐盐性。试验结果表明：盐胁迫后转基因株系的发芽率达到55%,比对照植株（Control plant,CK）高81%;转基因株系的鲜重、叶绿素含量均高于CK（P<0.05）;盐胁迫后转基因株系的存活率比CK提高75.0%~81.3%;盐胁迫明显抑制根系生长,但转基因植株的侧根数量比CK多4~5根（P<0.05）;盐胁迫后转基因株系体内的可溶性糖含量（P<0.05）和超氧化物歧化酶（P<0.01）,过氧化氢酶（P<0.01）,过氧化物酶（P<0.05）活性均高于CK,脯氨酸（P<0.01）和丙二醛（P<0.05）含量均低于CK。综上可得,MsLEA4-4基因参与了拟南芥体内的耐盐性调控,提高了转基因拟南芥对盐胁迫的耐受性。本研究为进一步探索MsLEA4-4在转基因紫花苜蓿盐胁迫中的功能及培育耐盐性紫花苜蓿品种提供依据。