Donor Plant Growth Conditions and Regeneration of Fertile Plants from Somatic Embryos Induced on Immature Zygotic Embryos of Sunflower (Helianthus annuus L.)

Immature zygotic embryos of the sunflower inbred line ‘Ha 300’, cultivated on a modified MS medium containing 6-benzyladenine and a high amount of sucrose, regenerated fertile plants via direct somatic embryogenesis. Plant regeneration from immature sunflower embryos is generally characterized by a relatively high experimental variability resulting from the interactions of multiple factors. We present here a study of some of the factors acting on the donor plants and their influence on the capacity to regenerate plants. Repeated experiments during a 2-year period with greenhouse-grown as well as field-grown plants led to the following conclusions: (i) The use of pesticides, unavoidable in the greenhouse, is compatible with routine regeneration of fertile plants, (ii) The plant growth retardants tested were useful for the production of healthy plants in the greenhouse and had no effect on the regeneration capacity.     

Detection and Elimination of Viruses in Callus, Somatic Embryos and Regenerated Plantlets of Grapevine

The distribution of some grapevine viruses in flower explants, embryogenic and non-embryogenic calli, single somatic embryos and plants regenerated from embryogenic cultures was investigated by RT-PCR and ELISA. Immature anthers and ovaries of the cultivars Grignolino infected by GRSPaV, GLRaV-1 and GVA, Müller-Thurgau infected by GRSPaV and GLRaV-3 and Bosco infected by GRSPaV were cultivated on media inducing indirect somatic embryogenesis. Viruses were detected both in anthers and ovaries. Four months after culture initiation 65.6% of tested calli were infected by at least one virus; high percentages of virus infection were found in calli originating from ovaries. No virus was detected in calli tested 8 months after culture initiation, as well as in single somatic embryos or in embryo-derived plantlets. Somatic embryogenesis confirmed its effectiveness in eliminating phloem-limited grapevine viruses. Regeneration of RT-PCR negative plantlets occurred even when at least a sector of the callus was still infected: the mechanism whereby somatic embryos are freed of some viruses could be related to the rapid proliferation of embryogenic cells within the callus or to the origin of the embryogenic callus from virus-free cells within the original explant.     

银杏幼胚离体培养再生植株的研究

 以佛手银杏幼胚为外植体, 研究了不同大小幼胚在不同培养条件下, 愈伤组织和胚状体的诱导发育情况。结果表明: 大于3 mm的幼胚, 在MK +NAA 1.0 mg·L ^{- 1} +BA 1.0 mg·L ^{- 1}培养基上胚状体诱导率最高, 达到53.6% , 最多的一块愈伤组织形成多达38个胚状体。暗培养不利于胚状体的发生。在MK培养基上添加10%椰汁对胚状体的生长发育有很好的促进作用, 有34.5%生长成苗。     

四川‘竹根姜’胚性细胞悬浮系的建立与植株再生

 选用四川省地方品种‘竹根姜’的茎尖组织建立了体细胞胚性细胞悬浮系, 并通过悬浮培养获得了姜再生植株。结果表明悬浮细胞干质量受pH影响。接种量和AgNO₃ 对悬浮细胞的生长都有影响:最佳接种量为1.0% , 最适AgNO₃浓度为6.0 mg·L ^{- 1}。胚性细胞悬浮系增殖后, 接种到前期筛选的成熟分化培养基上获得了再生植株。     

球根海棠离体培养的形态发生研究

 球根海棠外植体可在同一条件下同时诱导不定芽和胚状体, 经间接途径和直接途径的体细胞胚胎发生频率均很高, 体细胞胚为单细胞起源或多细胞起源。以再生嫩叶为继代中间体, 在较高温度(28 ℃) 下诱导, 可建立高效扩繁体系。纵切芽丛继代形成大量愈伤组织, 愈伤组织继代半年后几乎完全丧失分化能力, 发生激素驯化作用, 而且愈伤组织细胞染色体数目已发生广泛变异。     

德国核桃‘No.120''幼胚胚轴与子叶体细胞胚胎发生及其植株再生

核桃 (Juglansregia)‘No . 12 0’品系授粉 7～ 8周的幼胚胚轴与子叶在含不同植物生长调节剂组合的诱导培养基上培养 4周后转移至无植物生长调节剂的继代培养基上培养 ,不同发育时期的体细胞胚直接形成于胚轴与子叶表面。子叶与胚轴分别在KT 2mg/L BA1mg/L IBA 0 .0 1mg/L与 2 ,4 D 2mg/L KT 1mg/L的组合的诱导下体细胞胚胎发生频率最高。体细胞胚经 3～ 5℃低温处理 2个月 ,31.6%的体细胞胚能发育成正常生长的植株     

Potential,constraints and applications of in vitro methods in improving grain legumes

Grain legumes, the important constituents of sustainability‐based cropping systems and energy‐limited vegetarian diets have long been the subject of scientific research. Tremendous technological strides were made in the so‐called orphan crops, in terms of both varietal improvement and generation of basic information. Despite recalcitrancy and high genotype dependency, in vitro culture techniques such as organogenesis, in vitro mutagenesis, embryo rescue and in vitro gene transfer have been deployed for improvement of several grain legumes and these played an important role in introgression of desirable genes from related and distant species and creation of additional genetic variability. Stable and reproducible regeneration protocols resulted in the development of genetically modified chickpea, pigeon pea, cowpea, mungbean, etc., while embryo rescue was deployed successfully for recovery of interspecific recombinants, a few of them exploited for the development of commercial cultivars. Nevertheless, doubled haploidy witnessed limited success and protoplast regeneration and in vitro mutagenesis remained of academic interest. The present review focuses on the progress, achievements, constraints and perspectives of using in vitro technology in grain legume improvement.     

基于体细胞胚发生的细叶百合和兰州百合多倍体诱导及鉴定

以细叶百合（Lilium pumilum DC. Fisch.）和兰州百合（Lilium davidii var. unicolor）的鳞片和胚性愈伤组织作为诱导材料,用不同浓度秋水仙素以浸泡和混培两种处理方式进行多倍体诱导。以形态学观测、根尖染色体计数、气孔观测对多倍体植株进行倍性鉴定,分别获得了19株细叶百合四倍体植株和6株兰州百合四倍体植株,最佳处理方式为0.1%秋水仙素浸泡胚性愈伤组织24 h。使用ISSR对经体细胞胚发生的二倍体和多倍体再生植株进行遗传分析,发现两种百合二倍体植株遗传较稳定,而多倍体植株均发生遗传变异,其中细叶百合和兰州百合遗传变异率分别为15.48%和9.75%。     

Sequence analysis and expression pattern of AmLEA14 encoding a late embryogenesis abundant protein in Ammopiptanthus mongolicus

从沙冬青叶片EST文库中获得了胚胎晚期发生丰富蛋白（LEA）基因cDNA全长序列,命名为AmLEA14。序列分析表明该基因全长579bp,含有一个459bp编码152个氨基酸的开放阅读框,推测编码的蛋白质分子质量为16.6ku。二级结构预测显示此编码蛋白属于第4类LEA蛋白。系统发生分析表明,此编码蛋白与大豆LEA14蛋白（P46519.1）的亲缘关系最近。荧光定量PCR结果显示,AmLEA14的表达量在低温、干旱、盐胁迫条件下升高,但是主要在低温胁迫后期富集。推测AmLEA14基因可能在沙冬青抵御低温、干旱、盐胁迫中发挥作用,主要是参与沙冬青的低温防御机制。