影响梨叶片高频不定梢再生因素的研究

本文研究了基因型、基本培养基、植物生长物质、叶片接种方法及光、暗培养条件等因素对梨叶片不定梢再生的影响。基因型对叶片不定梢再生影响最大,安久和丰水的再生率最高,可达100%,其次为鸭梨,为80%,黄金梨的再生率最低,仅为27.3%。不同基因型其获得最高不定梢再生率所需附加的植物生长物质不同。TDZ比BA更有利于鸭梨叶片的高频不定梢再生,而BA更有利于西洋梨和砂梨叶片的不定梢再生。暗培养3周后再转光下培养比直接光培养有利于不定梢的再生,叶片接种方法对再生率无明显影响。     

Genetic control of somatic embryogenesis in alfalfa (Medicago sativa L. cv. Adriana)

Summary In alfalfa (Medicago sativa) regeneration is genotype-specific. In order to study the genetic control of somatic embryogenesis and to constitute a synthetic cultivar characterized by its high regeneration ability, 2 embryogenic plants selected from the cv. Adriana were selfed, intercrossed and also crossed in both directions with 5 non-embryogenic genotypes of the same cultivar.Progenies of all crosses were scored for their regeneration ability and results indicate that somatic embryogenesis is under the control of 2 dominant loci. However some non-embryogenic genotypes prevent regeneration when crossed with embryogenic ones and this characteristic is not under the control of a single dominant gene.When plants chosen for their capacity to regenerate within F₁ and S₁ progenies were freely intercrossed the regeneration efficiency dropped to 2% (1 plant out of 50). This result indicates that if the genetic background of the population is changed the regeneration is greatly affected and therefore some other mechanism could play a role in determining plant regeneration.     

青冈栎林分空间结构对林下更新的影响

林分空间结构会显著影响其林下幼树更新。为明确其影响林下幼树更新的关键因子,以湖南省青羊湖国有林场青冈栎林天然次生林为研究对象,测量幼树株数密度、树高、地径和冠幅,计算混交度、大小比数、开敞度指数以及聚集指数代表林分空间结构,评价林下幼树更新情况,并用灰色关联度分析方法探究两者间的相关关系。结果表明,青羊湖国有林场13块青冈栎林样地林下更新幼树有6种,分属于4科5属,分别是青冈栎、苦槠、多脉青冈、南酸枣、黄檀、马尾松,青冈栎在更新幼树中占明显优势,是优势树种,幼树重要值为63.09%。青冈栎林各林分空间结构指数与林下更新指数均大于0.5,关联度较高,综合所有林分空间结构指数对林下更新指数关联度的均值来看,对林下更新的影响由大到小为大小比数>开敞度>聚集指数>混交度,大小比数对青冈栎林下更新影响最为显著。在促进青冈栎天然次生林林下更新时,应以调整林木大小分化程度为主,综合考虑林木空间分布格局的调整方案。     

我国部分主推小麦品种组织培养再生能力评价

小麦细胞工程育种和基因工程育种存在强烈的基因型特异性, 从目前推广的优良小麦品种中筛选不同外植体再生能力强的基因型, 对于提高小麦生物技术育种效率和加速育成品种的生产应用具有重要意义。本研究以全国大面积推广的24个优良小麦品种和抗白粉病优良品系CB037为材料, 连续2年进行花药培养、幼胚培养和成熟胚培养, 统计愈伤组织诱导率、愈伤组织分化率和植株再生率, 分析、评价这些小麦品种(系) 3种外植体的组织培养再生性能。结果表明, 25个小麦品种(系)花药、幼胚、成熟胚的植株再生率分别为0~41.75%、2.25%~531.92%和3.24%~84.34%, 基因型差异显著; 组织培养再生能力以幼胚最强(119.79%), 成熟胚其次(36.23%), 花药最弱(4.91%)。CB037的3种外植体组织培养再生效率均最高, 轮选987、扬麦16、内麦836、科农199、新春6号、郑麦366、郑麦9023、新冬20、烟农19和川麦42幼胚培养植株再生能力表现较强, 新春6号、京冬8号、石麦4185、科农199和轮选987成熟胚培养植株再生率较高, 石麦4185和邯6172花药培养绿苗诱导率较高。小麦组织培养效率与基因型和外植体类型密切相关, 不同品种同一外植体再生能力差异显著, 同一品种不同外植体再生能力也存在显著差异, 并且3种外植体的组织培养再生能力不存在相关性。本研究筛选到不同外植体再生能力较好的优良小麦基因型, 可进一步用于小麦转基因育种和单倍体育种。     

大豆组织培养再生系统的研究进展

大豆因其愈伤组织难以分化、原生质体再生困难等因素,其再生体系一直不够完善。直到20世纪80年代初期,才成功建立了大豆的体细胞胚胎发生和器官发生再生系统。80年代末期原生质体培养获得突破。介绍了大豆器官发生再生系统、体细胞胚胎再生系统和原生质体再生系统的研究进展;对这3种再生系统进行遗传转化的优缺点作了比较;并提出了关于大豆遗传转化再生系统的几点展望。     

“济麦”系列品种成熟胚再生体系的优化研究

建立成熟的外植体再生体系,搭建有效的转化平台是实现小麦基因工程改良的重要途径。与未成熟胚相比,利用小麦成熟胚作为外植体不仅取材方便,而且不受环境和季节的限制。因此,建立小麦成熟胚再生体系是小麦组织培养发展的重要趋势。本研究利用"济麦"系列5个品种济南17、济麦19、济麦20、济麦21和济麦22,以适于组培的小麦品种Bobwhite作对照,研究了不同浓度2,4-D、玉米素(ZT)和激动素(KT)对小麦成熟胚的愈伤诱导和分化成苗的影响。结果表明,与对照品种Bobwhite相比,"济麦"系列品种具有更高的成熟胚出愈率,品种间差异显著,呈现出基因型效应。在相同2,4-D诱愈浓度条件下,不同品种间分化和成苗率差异极显著。"济麦"系列品种的成苗率均低于对照品种Bobwhite(13.55%),说明进一步优化愈伤的再生成苗能力是未来建立再生体系的关键。相同品种条件下,2,4-D诱愈浓度对出愈、分化和成苗率影响显著,当2,4-D诱愈浓度4mg/L时,除济麦20以外,其它五个品种均能分化成苗,其中济麦22成苗率约10.33%,接近Bobwhite水平。本研究为建立和完善"济麦"系列品种的成熟胚再生和遗传转化体系奠定了基础。     

植物原生质体再生细胞壁研究进展

原生质体作为单细胞且具有全能性,为研究植物细胞壁生物合成提供了独特的视角和模型。原生质体细胞壁再生过程复杂,涉及诸多细胞信号转导通路,应用普通的研究方法难以全面解析整个再生壁过程的分子网络。为充分了解植物原生质体再生壁内在特性,推动植物原生质体再生壁的分子机理进一步完善,笔者从原生质体再生细胞壁培养、细胞壁再生的细胞生物学证据、结合诸多组学技术研究细胞壁再生分子机理3个方面进行系统分析和归纳总结。分析了不同植物原生质体培养的主要影响因素如植物材料限制、培养基及培养方式的差异;归纳了2种常用染料应用于原生质体再生壁可视化研究的进展;总结了转录组、蛋白组及更为精细的核蛋白组及磷蛋白组技术在揭示细胞壁再生分子网络机理方面的应用和意义,并提出了对原生质体细胞壁再生机理深入研究的展望。     

草莓离体再生研究新进展

对草莓离体再生研究中的一些新进展作了介绍,从TDZ的高效利用、AgNO3、活性碳、重金属离子等附加物的作用、光对叶片再生的影响等方面进行了探讨,还对影响外植体器官发生的生理因子如活性氧和抗氧化酶与再生的关系进行了分析。当前存在的问题主要在于积累的技术要点多是经验性的,对再生机理知之较少,有必要对再生机理及遗传转化相关研究进行深入的、综合性的研究。     

“一步法”诱导三角紫叶酢浆草再生体系的形成

本研究以三角紫叶酢浆草叶片为材料,在现有研究的基础上,采用固定配比的6-BA（0.5 mg/L）和NAA（0.5 mg/L）,研究了KT、2,4-D对“一步法”诱导三角紫叶酢浆草再生体系的影响,研究表明MS+ 0.5 mg/L 6-BA + 0.5mg/L NAA + 1.0mg/L KT + 0.1mg/L 2,4-D的激素组合更能促进再生体系的形成,该激素组合可以诱导愈伤组织的形成,并诱导根及不定芽的发生及其增殖,三角紫叶酢浆草再生体系分化效率高,每块1cm2的叶片外植体平均可诱导成苗21.8棵。“一步法”诱导形成三角紫叶酢浆草再生体系方法能有效的降低工作量,而且具有再生苗诱导效率高的特点,有利于规模化繁育三角紫叶酢浆草。     

气象因子对北京地区不同秋眠等级苜蓿秋季刈割后再生高度的影响

为了研究气象因子对不同秋眠等级苜蓿秋后再生性能的影响,采用完全随机区组设计,研究了美国11个标准对照秋眠等级苜蓿品种秋季刈割后再生高度与气象因子的关系。结果表明：北京地区各秋眠等级的苜蓿在秋后刈割两周后再生高度便出现显著差异,各秋眠等级苜蓿植株再生高度的变化与三个气象因子的剩余效应较小,说明其他因子对苜蓿秋季刈割后植株再生高度的影响并不显著,因此可将积温、光照时数及降水量作为研究苜蓿秋眠性的主要影响因子。其影响程度依次为积温＞光照时数＞降水量。积温和光照时数对苜蓿秋季刈割后再生高度的影响最大。积温对再生高度的影响以直接效应为主,两者呈显著的正相关关系;光照时数和降水量均通过积温对再生高度产生正效应,而其对再生高度的直接影响为负效应。     

甘蓝型油菜带节子叶再生体系的建立及遗传转化应用

优良再生体系的建立是农杆菌介导植物遗传转化的基础。本研究建立以甘蓝型油菜含部分子叶节的子叶为外植体的再生体系,该再生体系为:切取5日苗龄甘蓝型油菜含部分子叶节的带柄子叶为外植体,置于添加3 mg/L6-BA的MS基本培养基中培养5天,外植体再生出丛生不定芽,不定芽再生率为100%。基于新建立的带节子叶再生体系,通过农杆菌介导,成功地将用p FGC5941载体构建的Bn TFL1基因干扰载体转入甘蓝型油菜中,从播种到得到生根抗性苗,整个转化周期只需要60~70天,较传统甘蓝型油菜以不带节子叶为外植体100~130天的转化周期大大缩短。     

苹果叶盘法基因转化中抗生素种类和浓度的筛选

以苹果品种嘠拉试管苗为试材，研究了卡那霉素（Km）、潮霉素（Hm）对继代苗生长和离体叶片再生的影响，以确定叶盘法基因转化的选择压和转化体的筛选浓度以确定苹果叶盘法基因转化中的选择压和抗性芽的筛选浓度；同时研究了不同浓度的头孢霉素（Cef）和羧苄青霉素（Carb）对苹果离体叶片再生的影响及对农杆菌EHA105和LBA4404的抑菌效果，以确定苹果叶盘法基因转化中合适的抑菌抗生素种类和浓度；以确定侵菌共培养后合适的抑菌抗生素种类和浓度。。结果证明表明：Km卡那霉素10mg/L、Hm4.0 mg/L完全抑制了叶片不定芽的分化，可作为苹果叶盘法转化时抗性芽的选择压； 在Km 50mg/L、Hm5.0 mg/L达到继代苗的半致死浓度，可作为基因转化后抗性芽苗的筛选浓度，10mg/L时完全抑制供试叶片不定芽的分化。；由于抗性芽再生的选择压与抗性苗筛选浓度相差较小，Hm相对于Km更适合作为苹果基因转化的选择标记；培养基中附加潮霉素的浓度为5mg/l继代苗不再分化生长，作为基因转化后抗性芽的筛选浓度，2.0mg/L时完全抑制供试叶片不定芽的再生，作为基因转化后抗性芽的筛选浓度。头孢青霉素在Cef 300mg/L时就能完全抑制农杆菌生长，对叶片不定芽的再生影响不大；而附加Carb羧苄青霉素则需400mg/L以上时才能抑制农杆菌LBA4404的生长，同时严重抑制了叶片再生。因此，宜选用Cef作为苹果基因转化的抑菌抗生素。     

大白菜再生体系影响因子的方差分析

用生物统计的方法对大白菜不定芽再生的影响因子进行分析。研究了基因型、激素和AgNO3对17种白菜子叶（带1－2mm子叶柄）外植体不定芽再生率的影响。三组方差分析说明，基因型、激素和AgNO3都在大白菜再生体系中起着重要的作用。基因型是大白菜高频再生的限制因素，它决定了相应的最适合的激素组成和AgNO3浓度；而对某一种基因型而言，激素组成和AgNO3浓度是建立高频再生体系的关键因素。     

Chromosomal regions associated with the in vitro culture response of wheat (Triticum aestivum L.) microspores

Generation of doubled haploid plants is a powerful tool in breeding, as homozygous individuals will be obtained directly from hybrids. However, genotype variability in regeneration efficiency of most European wheat (Triticum aestivum L.) varieties has limited its use in wheat. This study intended to identify quantitative trait loci (QTLs) for green plantlet regeneration from wheat microspore cultures. A QTL analysis using DArT markers was conducted based on a bi‐parental F₃ population, derived from a cross between the varieties Svilena and Jensen, which displayed markedly different capacity for plantlet regeneration. Two QTLs on chromosome 1B and 7B explained 53% of the variation in green plantlet regeneration. Furthermore, a collection of 94 European wheat varieties was genotyped and phenotyped. The microspore response level was low among western and northern European wheat varieties, and the positive QTLs found in the bi‐parental population were rare in the variety collection. Identification of the two QTLs enables introduction of high regeneration efficiency into wheat germplasm. Moreover, our results proved that the efficient regeneration observed for one variety could be crossed into modern winter wheat.     

Regeneration and transformation of cassava

A prerequisite for the development of a successful transformation system is the availability of efficient regeneration systems. Up to 1995 the only available regeneration system in cassava was an organized type of somatic embryogenesis. Transformation of these organized somatic embryogenic cultures with particle bombardment or Agrobacterium tumefaciens resulted in chimeric transformed embryos. However, the transformed sector was lost after repeated cycles of secondary somatic embryogenesis. After 1995 a less organized system of somatic embryogenesis was developed, so called friable embryogenic callus (FEC) and a system of adventitious shoot regeneration. The FEC regeneration system was combined successfully with particle bombardment. Selection of transgenic plants was based on either luciferase activity, or resistance to the aminoglycoside paromomycin or the herbicide phosphinothricin. Furthermore, protoplasts of FEC are able to regenerate into plants and can be transformed by electroporation. The adventitious shoot regeneration system was combined successfully with Agrobacterium tumefaciens. For this mature somatic embryos were cocultivated with Agrobacterium and cultured for adventitious shoot development. After selection based on the aminoglycoside geneticin or on hygromycin transgenic plants were formed. This revised version was published online in July 2006 with corrections to the Cover Date.     

THE STUDY ON INVESTMENT EFFECT OF OLD URBAN DISTRICT REGENERATION

The problem of old urban district regeneration has to be solved in process of ur-ban development.According to the practice of urban construction and the current situation of urbandevelopment in our country,solving this problem is very urgent. At present.there is fewer discussionon old urban district regeneration from its economic and social effect; In this paper.by means of an-alyzing old urban district regeneration and its investment effect, a set of criteria evaluating the in-vestment effect of old urban district regeneration is eastablished. For the first tirne, a new method. Set Statistics-AHP (The Analytic Hierarchy Process)-Entropy method is adopted to evaluate the in-vestment effect,and successfully solve two critical problems,individual criterion evaluation and crit-era' s weight.     

Concentration effects of dicamba on shoot regeneration in wheat

Shoot formation and regeneration rates of calli derived from immature embryos of three spring wheat varieties were investigated to evaluate the influence on wheat regeneration of different concentrations of dicamba (0.5, 0.1 and 0.02 mg/l) in the regeneration medium. It was found that dicamba concentrations much lower (0.02 and 0.1 mg/l) than the levels reported previously (1 and 0.5 mg/l) induced significantly more shoots per callus in all three varieties, while the enhancement in regeneration rate was variety dependent. Overall, a dicamba concentration of 0.02 mg/l was more favourable for wheat regeneration.     

Regengeration-Space:Study on Urban Regeneration in Vision of Space

Where to have regeneration and where to give it a priority are the basic questions need to answer in urban regeneration.Based on clarifying the defined method,this article aims at probing into the scope and the priority of regeneration-space.Then,a further survey of it from the angle of time and an analysis of the regeneration-space were carried out combined with the current situation of China.In order to reach a clear understanding of the space program of the city regeneration for a scientific decision for regeneration and raising the efficiency.     

Introgression of in vitro regeneration capability of Lycopersicon pimpinellifolium Mill. into recalcitrant tomato cultivars

The goal of this research was to study the introgression of the high regeneration capacity of Lycopersicon pimpinellifolium Mill line WV-700, in recalcitrant tomato cultivars (L. esculentum Mill cvs. Petomech, Santa Rita and VFN-8) using backcrossing. Hypocotyl explants of in vitro germinated seeds were cultivated in half strength MS medium supplemented with 5mg/l 6-BA to assess their shoot regeneration capacity. The apical shoot of the in vitro germinated seeds were cultured on a separate medium. Apical shoots from genotypes showing high regeneration rates were acclimated in a glasshouse and used as pollen donors for the next backcrossing. After four backcrossings, the material showed a similar mean fruit weight for the cultivated tomato and a high regeneration capacity similar to the wild species. It is shown that L. pimpinellifolium can be used with success as donor parent to introgress in vitro regeneration capacity to recalcitrant tomato cultivars. This revised version was published online in August 2006 with corrections to the Cover Date.     

Genetic analysis of tissue culture traits in barley cv. 'Lenins'

The barley cultivar ‘Lenins’ was found to be a genotype showing high shoot regeneration ability in cultures derived from immature embryos. Five cultivars different from ‘Lenins’ in shoot regeneration ability were reciprocally crossed with ‘Lenins’ and the inheritance of tissue culture traits was investigated. F₂ plants showed continuous distributions in callus growth and percentage of shoot regeneration, suggesting that these traits were controlled by polygenes. The F₂ population, derived from a cross between ‘Lenins’ and ‘6721′, showed a monogenic segregation for the number of regenerated shoots, and the segregation ratio fitted 1:2:1. Tissue culture traits of ‘Lenins’ were controlled by several genes, whereas the number of regenerated shoots related to the efficiency of shoot regeneration is controlled by one major gene.