基因型对春麦花粉植株诱导率影响的研究

花药培养中,不同基因型供体对于花粉植株的影响很大.几年来的研究结果表明,用花培品系做亲本配置杂交组合;不同世代的基因型,尤其是F_1代花药供体植株;亲本基因型在提高花粉愈伤组织出愈率、花粉绿苗分化及花粉绿苗率方面均有明显的效果.     

提高光敏感雄性不育水稻花粉单倍体育种效率及不育花粉植株育性转换特点的研究

比较了(光敏s/正常品种)F_1及F_2为供体亲本,对在花药培养时所获得的花粉植株中不育个体/全部花粉植株之比例的影响。结果表明,以F_1为供体亲本,在所获得的二倍体花粉植株(A_1)中,不育株(长日下)约占20%左右;而从F_2分离的不育株为供体亲本,相应的比例为90%左右。对获得不育的花粉植株而言,供体亲本经过F_2的选择,在花粉一代中可以提高育种效率3—4倍。指出,以培育光敏感雄性不育系为目的的花药培养,与一般育种之花药培养采用杂种F_1为供体亲本不同,不仅应对杂种F_2代在长日照条件下进行不育株的选择,而且应在短日照下对这种不育株作育性转换的双重选择。以这种个体作为花药培养的供体亲本,可以大大提高育种效率。 在长日照下表现不育的花粉植株的育性转换具多样性。来自同一组合的不育花粉植株在晚造(短日照)条件下,其花粉有的染色,频率高且稳定;有的虽然可变为染色,但频率不高或不稳定或二者兼有;有些却一直不为Ⅰ-KⅠ染色,或即使染色频率也在10%以下。这一结果与收集全国各地15个光敏核不育系在本昕同期种值条件下的反应十分吻合。这说明通过花药培养,从特定的组合培育出所需要的光敏/光温互作或温敏型的核不育系的可能性是存在的。     

离体再生能力的遗传调控和操作

许多农艺精典作物基因型难以离体再生,这样限制了利用如离体筛选或基因转移等组织培养技术进行育种来改良这些作物的能力。然而,一些现有研究表明植物的离体再生能力是受遗传调控的,可以通过操作使其表达该种特性。对一种春大麦(Hordeum spontaneum)品系间杂交产生的 F1和 F2代植株进行遗传分析,发现一种高花药培养反应性和两种较低反应性品系,慕尼黑科技大学的科学家 M.R.Islam 及其同事报道,这种花药培养反应性特性是可遗传的并以显性基因的方式起作用。研究还表明从花药培养物再生绿色植株的遗     

粳稻花药培养基因型差异的研究

对7个粳稻(Oryza sativa L. sub sp. kcng)品种按双列杂交方式配成的21个杂交组合进行花药培养,测定它们的愈伤组织诱导率和绿苗分化率。统计分析表明,基因型变异方差、一般配合力方差及特殊配合力方差均达到极显著水平。对亲本的一般配合力效应和组合的特殊配合力效应也进行了估算。研究表明,粳稻花药培养力是可遗传的,用高培养力的材料做亲本以提高杂种花粉植株的产量是有效的。此外还观察到供体植株在小孢子单核晚期前10天左右遇到特殊低温会使愈伤组织诱导率下降。     

水稻花粉植株的遗传学研究

对自1972年以来,通过花药培养所获得的近千株花粉植株的倍数性和部分来源于杂种F_1的花粉植株的纯合性及性状遗传规律进行了研究。结果观察到60％的花粉植株为能正常结实的二倍体。90％来自F_1的花粉植株的后代是整齐一致没有分离的,也即在遗传上是纯合的。其余10％来自F_1的花粉植株的后代或在性状上、或在能育性上、或同时在性状与能育性上发生分离。但性状分离一般都较简单,与F_2的分离有明显区别。对来源于同一杂交组合F_1的49个花粉植株的四个性状的重组与分离的规律,以同组合F_2植株为对照,进行了研究。结果两者表现基本相似。说明杂种F_1的各种小孢子发育为花粉植株的机率是相同的,没有发现在培养过程中不同基因型的花粉间有明显的竞争。 通过花粉单倍体育种已培育出二个粳稻新品种在生产上繁殖推广。     

植物提取物对小麦花粉愈伤组织器官分化的研究

小麦单倍体工作中,由于材料遗传差异大,绿苗分化率悬殊,有的根本不分化。为让较多组合得到绿苗,为小麦花培育种提供较多选育机会,我们试用小麦、玉米不同部位、不同浓度提取物作简化分化培养基,以(向阳4号×759-1)F_1为试材,诱导根芽分化形成完整植株方面取得初步结果。1、玉米、小麦不同提取物对愈组分化影响     

番茄花药培养研究进展及展望

在番茄育种中,利用花药培养产生单倍体植株,可以比常规育种方案较短时期固定和分析杂种优势的新遗传组合,建立纯合番茄品系,本文综述了国内外番茄花药培养研究概况及进展,并提供了展望。     

抗根肿病大白菜的花药培养

大白菜是中国北方的重要蔬菜之一,根肿病是危害大白菜生产世界性病害,利用花药培养可以大大加速抗根肿病大白菜杂交育种工作的进程。对33份抗根肿病大白菜品种(品系)进行花药培养,有24个品种诱导出胚,品种诱导率为72.7%,以东方皇冠×C11的诱导率最高,为1.86胚/蕾。对大部分基因型来说,在培养基中添加0.4 g/L谷氨酰胺的诱导效果最好。研究还发现生长在温室中的供体植株更容易诱导出胚状体。变绿的子叶型胚转到B5+0.2 mg/L BA+0.1 mg/L NAA+0.1%活性炭的胚分化培养基上可诱导生芽。     

提高光敏感雄性不育水稻花粉单倍体育种效率及不育花粉植株育性转换特点的研

比较了（光敏 s/正常品种）F1及F2为供体亲本,对在花药培养时所获得的花粉植株中不育个体／全部花粉植株之比例的影响,结果表明,以F1为供体亲本,在所获得的二倍体花粉植株（A1）中,不育株（长日下）约占20％左右;而从F2分离的不育株为供体亲本,相应的比例为90％左右,对获得不育的花粉植株而言,供体亲本经过F2的选择,在花粉一代中可以提高育种效率3－4倍,指出,以培育光敏感雄性不育系为目的的花药培养,与一般育种之花药培养采用杂种F1为供体亲本不同,不仅应对杂种F2代在长日照条件下不育株的选择,而且应在短日照下对这种不育株作育性转换的双重选择,以这种个体作为花药培养的供体亲本,可以大大提高育种效率。在长日照下表现不育的花粉植株的育性转换具仍多样性,来自同一组合的不育花粉植株在晚造（短日照）条件下,其花粉有的染色,频率高且稳定;有的虽然可变 染色,但频率不高或不稳定或二者兼有;有些却一直不I－KI染色,或即使染色频率也在10％以下,这一结果与收集全国各地15个光敏核不育系在本所同期种植条件下的反应十分吻合,这说明通过花药培养,从特定的组合培育出所需要的光敏／光温互作或温敏型的核不育系的可能性是存在的。     

第三次小麦花药培养经验交流会召开

1984年4月10日至15日,在洛阳市召开了全国小麦花药培养第三次经验交流会,并对洛阳市农科所新选育的小麦品种“豫花一号”进行了鉴定。到会的有全国15个省市34个单位的63名代表。收到论文20余篇,会上作学术报告19篇。内容包括改进培养技术;供体植株的基因型对花药培养的影响及遗传规律;利用单倍体植株研究染色体工程创造新的遗传原始材     

Wheat anther culture: effect of genotype and environmental conditions

Twenty-two cultivars and lines of winter and spring wheat (Triticum aestivum L.) were studied, most for the first time, for their anther culture response. The response was genotype dependent. Plants grown in the field gave higher callus induction frequency than those grown in the greenhouse and the controlled environment chamber. Donor plants grown in a season of low drought stress as compared to a season of severe drought stress resulted in a higher frequency of callus induction. Spherical microcalli were observed in two wheat genotypes in some of only those anthers that were placed with only one loculus in contact with the medium. Wheat lines that were more responsive to anther culture were identified.     

不同组织培养途径对小麦再生能力的研究

从现在推广的小麦优良品种和有苗头的新品系中选用10个小麦基因型品种进行组织培养,从愈伤组织诱导率、绿苗分化率等方面比较了幼穗培养、花药培养、幼胚培养三种培养方式的培养效果。结果表明,幼胚培养效果最好,基因型间差异小,都能获得足够数量的再生植株。幼穗的培养效果最差,愈伤组织分化生根和绿芽十分容易,但分化成完整植株则较为困难。花药培养在基因型间差异非常明显而且有较多白化苗。此外,本研究还分析了影响小麦再生能力的因素,建立了一套高效、可靠的小麦组培再生系统,为小麦的转基因技术提供优良的受体材料。     

Anther culture as an effective tool in winter wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) breeding

The aim of this study was to determine the effect of genotype and induction medium in anther culture of wheat (Triticum aestivum L.). Ten F1 winter wheat genotypes were tested in anther culture (AC) to compare the two most frequently applied induction media (W14mf and P4mf). Androgenesis was induced during the treatment of each tested genotypes and green plants were produced from them using both media. Based on statistical analysis, the genotypes significantly influenced (at the 0.001 probability level) the efficiency of AC (embryo-like structures (ELS), albinos, green plantlets and transplanted plantlets) and the media also had a significant effect on the number of ELS and albino plantlets. Both media can be used for AC in wheat doubled haploid (DH) plant production. The production of ELS and green plantlets was higher in P4mf medium (48.84 ELS/100 anthers, 4.82 green plantlets/100 anthers) than in W14mf medium (28.14 ELS/100 anthers, 4.59 green plantlets/100 anthers). However, the green plant regeneration efficiency of the microspore-derived structures was 16.9% when using W14mf medium, while this value was 9.6% in the case of ELS induced with P4mf medium. The application of W14mf medium thus proved to be time- and labour-saving medium in the large-scale production of DH wheat plants. In our experiments, 267 DH plants were produced for our winter wheat breeding program. The spontaneous rediploidization rate was 32.72%.     

Genetic analysis of anther culture response in wheat using aneuploid,chromosome substitution and translocation lines

Summary Marked effects of genotype on wheat anther culture response have been observed. Genetic factors have been recognised to be one of the major contributors to in vitro responses of cultured wheat tissues. In wheat anther culture, embryo induction, plant regeneration and albina/green ratio have been determined to be heritable traits. Using Chinese Spring (CS) monosomic 1D, single chromosome substitution lines of chromosome 5B or chromosome arm 5BL from Chinese Spring into six varieties, and F₁ hybrids heterozygous for the 1B chromosome structure (1BL-1BS/1BL-1RS), the anther culture response was studied: genes on CS1D chromosome and 5BL chromosome arm increases the embryo frequency; gene(s) involved in regeneration ability are located on the 1RS chromosome arm; a gene increasing albina frequency is located on Chinese Spring 5B chromosome. Our results support the fact that without gametic selection, a differential development occurred from the particular classes of microspores carrying genes for higher regeneration ability. Moreover, in some crosses, a few genes with major effects were involved in determination of anther culture response.     

Effect of induction medium pH and maltose concentration on in vitro androgenesis of hexaploid winter triticale and wheat

Low levels of in vitro androgenesis limit the utility of anther culture as a routine tool for the improvement of triticale. The objectives of this research were to determine the effect of induction medium modifications on the embryoid induction (EI) and green plant regeneration (GPR) of three hexaploid winter triticales and a hexaploid winter wheat. Medium modifications were factorial combinations of gelling agents, pH and maltose concentration. There was a significant difference in the response of wheat and triticales. An induction medium pH of 4.8 vs. 5.8 led to higher EI percentages in all cases and in Ficoll containing medium to a significantly higher GPR in the wheat, but it had no effect on the response of the triticales. Maltose at 0.26 M vs. 0.21 M increased the EI of both the triticales and the wheat but led to higher GPR only in the case of wheat. Averaged over genotypes, induction media with agar and Ficoll 400 were superior to the liquid form. In terms of GPR the wheat gave the highest response: 9.1%. The GPR percentages for the triticales ranged from 1.4 to 7.1. Genotype specificity must be overcome if anther culture is to be routinely used in diverse arrays of triticale germplasm.     

Analysis of quantitative trait loci which contribute to anther culturability in rice (Oryza sativa L.)

Anther culturability of rice is significantly different between indica and japonica varieties. A doubled haploid (DH) population was established via anther culture of an indica/japonica hybrid on SK₃ medium, which had been shown particularly suitable for anther culture of indica/japonica hybrids. For analyzing the quantitative trait loci (QTLs) responsible for anther culturability, anthers of the DH lines were again cultured with SK₃ medium and parameters for four traits representing the anther culturability were surveyed and analyzed with the molecular map constructed from the same DH population. The parameters for four major traits were as follows: callus induction frequency (CI), green plantlet differentiation frequency (GPD), albino plantlet differentiation frequency (APD), and green plantlet yield frequency (GPY). All four traits displayed continuous distributions among the DH lines. The correlation coefficients between these traits were also tested and showed that there was no relationship between callus induction and green plantlet differentiation frequencies, but both showed strong positive correlation with the frequency of green plantlet yield. For callus induction frequency, five QTLs were identified on chromosomes 6, 7, 8, 10 and 12. Two QTLs for green plantlet differentiation frequency were located on chromosomes 1 and 9. There was a major QTL for albino plantlet differentiation frequency on chromosome 9. No independent QTL was found for green plantlet yield frequency. The results may be useful in the selection of parents with high response to anther culture for rice haploid breeding and in the establishment of permanent DH populations for molecular mapping.     

The genetic and molecular characterization of pollen-derived plant lines from octoploid triticale x wheat hybrids

Six doubled-haploid (DH) lines, derived by anther culture from octoploid triticale x wheat hybrids, were characterized using cytological, biochemical and molecular techniques. Lines varied in their wheat and rye genome composition, and were either wheat-rye chromosome multiple addition lines or had spontaneous substitutions and/or wheat-rye translocations. Most of the lines contained a pair of 4R chromosomes, whereas 1R or 7R were present in others. The results are similar to those previously obtained with hexaploid triticale x wheat crosses and indicate that it is possible to produce alien (wheat/rye) addition, substitution, and translocation lines directly from the anther culture of intergeneric hybrids.     

Effect of genome, induction medium and temperature pretreatment on green plant production in durum ( Triticum turgidum var. durum ) x bread wheat ( Triticum aestivum L. em Thell) crosses

The recalcitrancy of durum wheat (Triticum turgidum var. durum) to anther culture, was attempted to be overcome by transferring the responsible genes form bread wheat B-genome to the respective on durum wheat, determining an appropriate induction medium and clarifying the necessity of cold pretreatment. For this, three durum wheat cultivars were crossed to two bread wheat (Triticum aestivum L. em Thell) cultivars. The resulting F₁ plants and their original cultivars were grown in the field and anthers at the appropriate microspore stage were cultured on potato-2 and W₁₄ media with and without low temperature pretreatment. No green plants were produced from the parental durum wheat cultivars. In contrast, green plants were produced from the F₁ plants. The best results in three of the four F₁ hybrids were recorded when potato-2 was used as induction medium. A more variable response of the examined genotypes was noticed with respect to temperature pretreatment. Regarding green plant production, a negative effect of cold pretreatment was observed in two of the F₁ hybrids when they were cultured on potato-2. Chromosome counts on root tips from the resulting green plants revealed that they all carried D-genome chromosomes. The last observation could suggest that D-genome chromosomes are necessary for anther culture response in wheat. Yet, the production of one green plant with 15 chromosomes may indicate that the development of extracted durum genotypes from bread wheat genotypes with good response to in vitro anther culture might be possible. Further work however, is needed for this to be verified.     

Induction of in vitro androgenesis in anther and isolated microspore culture of different spelt wheat (<Emphasis Type="Italic">Triticum spelta</Emphasis> L.) genotypes

This is the first report on isolated microspore culture—derived spelt wheat. The efficiency of anther- and isolated microspore was compared using four genotypes (‘Franckenkorn’, ‘GK Fehér’, ‘Mv Martongold’, ‘Oberkulmer Rotkorn’). In anther culture, genotype dependency was observed, and cold pre-treatment enhanced the efficiency of the method. In isolated microspore culture, the ovary co-culture supported the development of embryo-like structures. The presence of growth regulators (0.5 mg/l 2,4-D and 0.5 mg/l kinetin) were not essential for the induction of androgenesis, but these increased the production of embryo-like structures, green and albino plantlets. The low plant regeneration rate and high number of albinos hinder the practical application of isolated microspore culture while anther culture was efficient for in vitro green plantlets production in spelt wheat. The mean of green plantlets production was 41.45/100 anthers (from 20.93 to 83.07 depending on genotype). The phenomenon of albinism was mitigated in anther culture (3.48 albinos/100 anthers). Altogether, 1720 anther culture—derived green plantlets were produced from the four genotypes.     

Screening wheat genotypes for high callus induction and regeneration capability from anther and immature embryo cultures

Plant regeneration via tissue culture varies with the genotype and is an important factor in establishing cell selection and genetic transformation systems. To select genotypes – especially Japanese ones – with a high regeneration capability, we screened 107 wheat genotypes (78 domestic, 29 foreign) for callus induction and regeneration capability from anther and immature embryo cultures. For anther culture, 83 of 107 genotypes tested induced calli and 45 regenerated plants. Only 9 genotypes, however, produced green plants, 25 produced only albino plants, and 11 produced both green and albino plants. Glennson 81 was the highest in callus induction, followed by Orofen, Danchi–komugi and Chris. The genotypes with a relatively high regeneration capability were Framala 80 at 24% and Glennson 81 at 19%, these two genotypes produced only green plants. For immature embryo culture, 97 genotypes showed a 90% callus induction rate and 74 genotypes regenerated plants. Very few genotypes produced albino plants. The genotypes with a high regeneration capability were Genaro 81 at 90%, Chinese Spring at 80%, and Norin 75 at 75%. This revised version was published online in June 2006 with corrections to the Cover Date.