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。     

海边香豌豆胚性愈伤组织的诱导和体细胞胚发生

将生长１４ｄ的海边香豌豆（Ｌａｔｈｙｒｕｓ ｍａｒｉｔｉｍｕｓ（Ｌ．）Ｂｉｇｅｌ）无菌苗下胚轴切成０．５ｃｍ左右的片段,置于含有１ｍｇ／Ｌ２,４－Ｄ,０．５ｍｇ／Ｌ ＢＡ和０．５％ＮａＣｌ的ＭＳ培养基中,２８ｄ后诱导出胚性愈伤组织。将其转入含有适当浓度２,４－Ｄ的ＭＳ培养基上,又２８ｄ后可得到大量球形胚和心形胚以及极少量鱼雷胚和子叶胚。诱导体细胞胚适合的２,４－Ｄ浓度为０．５ｍｇ／Ｌ。较高浓度的２     

杉木未成熟胚胚性愈伤组织诱导影响因素探析

该研究从基因型、6-BA浓度、外植体接种方式和合子胚发育阶段等方面,分析杉木未成熟胚胚性愈伤组织诱导的影响因素。结果表明:基因型、6-BA浓度、外植体接种方式和合子胚发育阶段均对胚性愈伤组织诱导频率有不同程度影响。6种基因型中,有3种基因型诱导出胚性愈伤组织,其中基因型S18胚性愈伤组织诱导频率最高,为11.7%。6-BA浓度在1.0~1.5 mg·L~(-1)范围内时,基因型S18的胚性组织诱导频率较高。以在去皮种子的一端切开一个小口的接种方式为最优,将合子胚剥出的方式易造成合子胚褐化死亡,将未剥皮的种子切开一个小口后直接接入培养基的方式不利于愈伤组织生成。适合胚性愈伤组织诱导的合子胚发育阶段为受精至胚器官分化阶段,合子胚进入成熟阶段后不利于胚性愈伤组织诱导,合子胚易生长成完整植株。     

兴安落叶松胚性愈伤组织诱导影响因子的研究

以成熟和未成熟合子胚为外植体,研究影响兴安落叶松（Larix gmelinii）胚性愈伤组织诱导的几种主要因子。结果表明兴安落叶松合子胚带胚乳培养有利于胚性愈伤组织的诱导;内蒙沙地种源成熟合子胚的诱导率显著（p<0.05）高于加格达奇山地种源;冷藏处理可以提高成熟合子胚胚性愈伤组织的诱导率;不同发育时期的未成熟合子胚的诱导率存在显著差别（p<0.05）,其中以子叶初期合子胚（7月5日）诱导率最高;2,4-D对胚性愈伤组织诱导的影响较大,且与BA、KT存在一定的协同作用;S培养基比DCR和MS培养基更有利于胚性愈伤组织的诱导;培养基中琼脂含量为4 g·L^-1时,诱导率较高。     

速生湿地松良种胚性愈伤组织诱导与增殖

为使速生湿地松良种快速大规模繁殖,对其胚性愈伤组织进行诱导和增殖优化研究.该文以1代湿地松种子园中10个速生湿地松优良无性系(基因型)的未成熟合子胚为外植体,系统研究基因型、合子胚发育阶段、基本培养基、植物生长调节剂种类和浓度等不同因子对胚性愈伤组织诱导效率的影响,探讨胚性愈伤组织的增殖条件.结果表明:基因型、合子胚发...     

柑桔胚性愈伤组织的诱导、保存和利用

对近３０年来柑桔胚性愈伤组织诱导及其影响因素,保存,利用和存在的问题以及今后的研究方向进行了概述。     

安祖花胚性愈伤组织诱导及植株再生研究

为构建安祖花(Anthurium andreanum)胚性愈伤组织再生体系,以3个盆栽品种幼嫩叶片和叶柄为外植体,分析了基本培养基、植物生长调节剂组合和培养条件等因素的影响。结果表明,安祖花胚性愈伤组织诱导的最佳培养基为改良MS3+1.5 mg L–1 2,4-D+0.5 mg L–1 KT+4%蔗糖+2%葡萄糖+0.25%Phytagel,且胚性愈伤组织诱导能力差异显著,表现为?粉冠军??罗宾奴??冠军?和叶片叶柄,其中?粉冠军?叶片的胚性愈伤组织诱导率可达57.9%。胚状体分化的最佳培养基为1/2改良MSa+2%蔗糖+0.25%Phytagel,其中?粉冠军?叶片诱导的胚状体分化率可达31.6%,且在光、暗下分化率的差异不显著。分化苗移栽后的成活率可达100%。     

ABA、NAA诱导水稻胚性愈伤组织的研究

ABA和NAA联合使用能有效地诱导水稻原生质体再生的愈伤组织向胚性发展。通过液体浅层培养由原生质体得到的愈伤组织,在含ABA和NAA的N_6培养基上培养一段时间,可以诱导原来呈非胚性状态的愈伤组织形成胚性愈伤组织,并在含ZT的N_6分化陪养基上产生绿点。通过对这两种愈伤组织的生化分析,表明二者在游离氨基酸、DNA、RNA、核酸及蛋白质含量等方面,特别是SDS-PAGE谱带存在明显的差异,其细胞的形态与结构也有显著差别,其中经ABA NAA诱导后的愈伤组织其细胞形态与结构特征与来源于种胚的胚性愈伤组织基本类似,所分析的生化指标也大多数相近。结果表明,ABA和NAA联合使用得当,能促进形成胚性愈伤组织。     

水稻愈伤组织内部胚性细胞的形成及发育

发育成胚状体的水稻愈伤组织表面胚性细胞,在继代过程中被非胚性细所包围,渐渐形成内部胚性细胞。共形态结构与表面胚性细胞相同,但周围缺乏表面细胞下面的一层至几层含丰富的淀粉粒的细胞,内部胚性细胞形成团后行然以外形成突起,产生根冠原,逐渐形成根而突出愈伤组织,内部胚性细胞可向四周同时但不同步形成根冠原,未见芽或胚状体从内部胚性细胞产生,推测胚性愈伤组织失支胚胎发生能力及分化能力可能部分与胚性细胞部分裂或分裂不旺盛、,而非胚性细胞分旺盛从而使胚性细胞被包围,。稀释有关。     

ABA,NAA诱导水稻胚性愈伤组织的研究

ＡＢＡ和ＮＡＡ联合使用能有效地诱导水稻原生质体再生的愈伤组织向胚性发展。通过液体浅层培养由原生质体得到的愈伤性发展。通过液体浅层培养由原生质体得到的愈伤组织,在含ＡＢＡ和ＮＡＡ的Ｎ８培养基上培养一段时间,可以诱导原来呈非胚性状态的愈伤组织形成胚性愈伤组织,并在含ＺＴ的Ｎ６分化培养基上产生绿点。通过对这两种愈伤组织的生化分析,表明二者在游离氨基酸、ＤＮＡ、ＲＮＡ、核酸及蛋白质含量等方面,特别是ＳＤＳ     

不同种源花榈木苗期生长及生理特性比较

为了解不同种源花榈木在贵阳的生长特性和差异,该文通过对10个种源地花榈木进行育苗试验,测定其两年生实生苗的苗高、地径、生物量、叶片光合参数、光合色素、硝酸还原酶活性、硝态氮含量和根系活力,并进行差异性分析。结果表明:(1)10个种源花榈木净光合速率、气孔导度、胞间CO₂浓度、蒸腾速率和水分利用效率差异显著(P<0.05),表明不同种源花榈木光合特性及光能利用效率具有较大差异,浙江杭州和浙江永康花榈木是具有较高光合生长潜力的种源。(2)种源间的叶绿素含量、硝酸还原酶、硝态氮、根系活力存在显著差异,福建建瓯种源的叶绿素a、叶绿素b含量和叶绿素总量最高,能够将光合原初反应过程中积蓄的光能进行高效地传递,促进碳的同化; 贵州花溪种源硝酸还原酶活性最大,硝态氮含量最高,对氮元素的利用能力较强,能够促进植物蛋白质、氨基酸和叶绿素等的合成; 贵州望谟种源根系活力最大,吸收养分的能力强。(3)各种源间苗高、地径和生物量的分配存在显著差异,浙江杭州种源的植株枝叶繁茂、根系发达,生长表现好,安徽黄山种源的植株矮小,生长表现较差; 浙江杭州种源将生物量更多分配在根和叶,提高其根系吸收养分和叶片获取光能的能力,安徽黄山种源总体生物量积累最少,长势最差。(4)通过主成分分析法对各种源的花榈木适应性进行综合评价,结果显示浙江杭州种源>贵州黎平种源>浙江永康种源>贵州望谟种源>福建建瓯种源>贵州凯里种源>贵州石阡种源>贵州花溪种源>贵州平塘种源>安徽黄山种源。综上结果表明,浙江杭州、贵州黎平和浙江永康种源花榈木对贵阳地区立地环境具有较强的适应能力和生长潜力。     

Influence of some exogenous amino acids on the production of maize embryogenic callus and on endogenous amino acid content

The effects of four exogenous amino acids (proline, glycine, asparagine and serine) on the production of maize embryogenic callus and on its endogenous amino acid content have been investigated. For this purpose, an established embryogenic line of Type 1 callus from the inbred W64Ao2 has been used. From the results it may be concluded that a concentration of proline exceeding 6 mM is negative for the production of embryogenic callus. When proline is eliminated from the medium, other amino acids tested in certain concentrations yield a percentage of embryogenic callus production that exceeds or equals that of proline. The endogenous free proline content in embryogenic callus is significantly higher than that in non-embryogenic callus regardless of proline presence in the medium. The only exception are the glycine-containing media, in which endogenous free alanine of embryogenic callus increases at the expense of endogenous free proline. This study suggest a positive role of endogenous free proline or alanine accumulation in the embryogenic callus production which might be related to an adaptation to the metabolic changes produced by in vitro culture and embryogenesis induction. Furthermore, these results indicate that treatments with amino acids that are different from proline can be used to improve the efficiency of embryogenic callus production from well established maize callus cultures.Abbreviations Ala alanine - Asn asparagine - 2,4-d 2,4-dichlorophenoxyacetic acid - EC embryogenic callus - nEC non-embryogenic callus - Gaba gamma-aminobutyric acid - Glu glutamic acid - Gly glycine - Pro proline - Ser serine     

AgNO3对橡胶树花药愈伤组织形态及体胚发生的影响

橡胶树的花药愈伤组织在长期继代过程中,胚性易下降甚至丧失;而AgNO3作为乙烯活性抑制剂,被广泛应用于植物组织培养中.该研究以继代培养4 a以上的热研7-33-97花药愈伤组织为材料,在继代培养基中添加2.5 mg·L-1 AgNO3预培养35 d后,观察预培养前后愈伤组织表形及其细胞形态的变化,并设计不同浓度AgNO3及不同处理时间对其进行体胚诱导,90 d后分别统计胚状体总数和正常胚数.结果表明:浅黄色质地柔软的愈伤组织在含AgNO3的培养基上预培养后能转变成鲜黄色易碎愈伤组织,在倒置显微镜下前者大多表现为不规则多边形,细胞内含物较稀薄;而后者则呈圆形或椭圆形,细胞内含物丰富,属于典型的胚性细胞.在体胚诱导的第1个月添加5 mg·L-1 AgNO3能显著促进体胚的发生,AgNO3浓度升至10 mg·L-1时体胚发生受到抑制,且畸形胚的形成率显著增加;在含5 mg·L-1 AgNO3的培养基中培养2个月以上,体胚发育明显受阻,大部分形成畸形胚.该研究结果在一定程度上恢复了橡胶树长期继代花药愈伤组织的胚性能力,并提高了其体胚发生频率,为橡胶树花药胚性愈伤组织长期继代培养过程中胚性的保持提供了参考.     

Differential gene expression in embryogenic and non-embryogenic clusters from cell suspension cultures ofCoffea arabica

Somatic embryogenesis (SE) is a very useful system for studying the differentiation process in plants and involves gene regulation at several levels. During SE induction in Coffea arabica cv. Catura Rojo two types of cell clusters, embryogenic (EC) and non-embryogenic (NEC), were observed. The goal of this work was to compare the most relevant characteristics between EC and NEC for a better understanding of the mechanism driving SE. Morphohistological observations indicated a correlation between the morphological features of clusters and their embryogenic competence. On the other hand, no variation at the DNA level, studied by AFLP, were found to explain the disparity in embryogenic competence of clusters, but gene expression, observed by RNA differential display, and SDS-PAGE showed differences that can explain that disparity. Our results lead us to propose that differential gene expression can modulate the embryogenic capacity of coffee cells and that the number of genes turned off in somatic cells to allow for the change from a somatic to an embryogenic state, is higher than those genes that are turned on.     

Endogenous hormone concentrations and embryogenic callus development in wheat

Immature zygotic embryos of two wheat (Triticum aestivum L.) genotypes, known for their different ability to generate embryogenic callus, were used as initial explants to establish callus cultures. Embryogenic and non-embryogenic calluses were obtained from the competent genotype (`Combi'), while only non-embryogenic callus was produced by the incompetent one (`Devon'). The morphogenetic competence of each callus type was evaluated by transferring some segments to regeneration conditions. The endogenous hormone concentrations (free indole-3-acetic acid [IAA], abscisic acid [ABA], gibberellins ₁, ₃ and ₂₀ [GAs], zeatin/zeatin riboside [Z/ZR] and N ⁶[²-isopentenyl] adenine/ N ⁶[²-isopentenyl] adenosine; [iP/iPA]) of the initial explants were determined by means of radio-immunoassay and showed that the only difference was the higher concentration of ABA found in the embryos of the most competent genotype; whose embryos showed a reduced rate of precocious germination. When analysing the endogenous hormone concentrations in the various callus types generated in each genotype, it was found that only differences in the free IAA concentrations were associated with variations in the morphogenic properties of the calluses. Higher concentrations of endogenous free IAA were typical of embryogenic callus cultures. It was also observed that a loss in the embryogenic competence of the calluses, due to a prolonged time of culture, occurred concomitantly with a reduction in free IAA concentrations, practically to the concentrations found in the non-embryogenic calluses.     

Modification of the embryogenic response of Coffea arabica by the nitrogen source

Somatic embryogenesis was induced in coffee from in vitro cultured plants as starting material and the faster response obtained allowed lines from selected plants to be generated more quickly. In contrast to other systems, where embryos take 2 or 3 months to develop, globular embryos were obtained after 3 weeks. The optimum nitrogen concentrations for embryogenesis were between 3.75 and 15 mM nitrogen with a nitrate/ammonium molar ratio of 2:1 or 1:2.     

杉木和米槠凋落叶DOM对土壤碳矿化的影响

DOM(Dissolved organic matter)是土壤微生物呼吸的重要底物,凋落物淋溶的DOM对土壤碳矿化具有重要影响。选择中亚热带地区具有代表性的杉木(Cunninghamia lanceolata)和米槠(Castanopsis carlesii)凋落叶作为研究对象,通过两个月的短期室内培养,把不同凋落叶浸提出的DOM添加到培养瓶中,定期测定土壤碳矿化速率,计算土壤碳累积矿化量,探讨两种等浓度等量DOM添加对土壤碳矿化的影响,并分析DOM化学性质在土壤碳矿化过程中的重要性。结果表明:米槠凋落叶浸提得到的DOC(Dissolved organic carbon)和DON(Dissolved organic nitrogen)浓度均显著高于杉木凋落叶的(P0.05),而杉木凋落叶浸提得到的DOM的UV吸收值(SUVA_(254))和HIX(Humification index)均显著低于米槠凋落叶的(P0.01)。添加等浓度等量杉木和米槠凋落叶DOM到土壤中均显著增加了土壤碳矿化速率,在第1天内分别比对照高198%和168%,3d后下降到61.8%和44.1%,14d后基本处于平稳状态,表明外源有机物添加对土壤碳矿化的前期影响较大。培养过程中,添加杉木和米槠凋落叶DOM的土壤碳矿化累积量均能采用双因素指数模型进行拟合(r~2=0.99),但添加两者凋落叶DOM后土壤碳矿化累积量没有显著差异。     

不同环境因子及破壳处理对锥栗种子萌发特性的影响

锥栗(Castanea henryi)是我国南方重要的木本粮食树种和特色果树资源,具有很高的食用及药用价值。为探明最适宜锥栗种子萌发的条件,该研究以当年采收的锥栗种子为材料,探讨了温度、光照、基质、基质含水量以及破壳处理等对锥栗种子萌发的影响,为锥栗种苗繁育提供理论基础。结果表明:(1)锥栗种子萌发的最适温度是25℃,其最终萌发率为92.67%,在其他温度条件下,锥栗种子的萌发率有所下降,但与25℃条件下的萌发率无显著差异。(2)黑暗条件可缩短锥栗种子发芽时间,明显提高种子的萌发率和发芽势,但黑暗条件下的种子胚根长度显著小于周期性光照条件下的胚根长度。(3)锥栗种子在泥炭土中的萌发率和发芽势均高于在河沙中的萌发率和发芽势,且泥炭土中种子胚根长度显著长于河沙中的种子胚根长度。(4)基质含水量对锥栗种子萌发率和发芽势无显著性影响。(5)种皮的机械阻碍是限制锥栗种子萌发的因素之一,破壳处理后,锥栗种子的发芽势和胚根长度与未破壳相比差异显著,实际生产中剪破种壳能加快种子的萌发进程。