不同世代小麦花药培养的遗传效应分析

实验用4个小麦杂交组合的6个不同世代群体为材料分别进行花药培养,对各材料出愈率、绿苗率、白苗率和白苗/绿苗比4个性状的世代平均值进行多元回归分析。结果表明:各组合出愈率存在明显的显性效应和显性×显性互作;绿苗率以加性效应和显性效应为主;白苗率则主要受上位性效应的影响.从而提出在小麦花药培养中注意利用杂种优势可以提高出愈率和绿苗率;选择一般配合力高的亲本杂交可以提高绿苗率,但对白苗率无多大影响。本文对同一性状不同组合、同一组合不同性状的遗传构成以及遗传因素对花粉白苗的影响等问题进行了讨论。     

基因型和环境条件对小麦花药培养效果的影响

为进一步提高小麦花培育种效率,明确花药培养力的遗传控制基础,以11个小麦品种及其组配的20个F₁杂种为材料,探讨了基因型、培养基和环境条件对愈伤组织诱导率的影响。在W14D、W14gD、W14GD培养基上,Alondra、Verry、石4185、新春9号和百农3217的花药易被诱导产生愈伤组织,诱导率为25.3%~51.9%,其中石4185是目前公认的花培育种优良亲本,新春9号为新发现的优良花培基因型。以宁春4号配制的部分F₁杂种的愈伤组织诱导率较高,大多数组合高于10.0%,表明宁春4号与供试品种间具有较高的花药培养配合力。小麦花培育种技术要求亲本之一具有较高的花药愈伤组织诱导率或较高的花药培养配合力。小麦花药培养力的遗传控制复杂,表现为数量性状遗传,亲本花药培养力很高,其F₁组合花药培养力不一定很高,这与双亲配合力有关。小麦花药培养中,供体植株生长和愈伤组织诱导的适宜条件为较长的营养生长期、适宜的前期(分蘖期)温度和较高的中期(拔节后期)温度。在添加低浓度生长素和葡萄糖的液体培养基中发现小麦花药直接成苗现象,2,4-D诱导花药直接成苗效果优于Dicamba。随着年度间气候升高的影响,相同基因型花药愈伤组织诱导率呈现增加趋势。     

Addition of Colchicine to Wheat Anther Culture Media to Increase Doubled Haploid Plant Production

Chromosome doubling is critical for obtaining doubled-haploid plants from wheat (Triticum aestivum L.) anther culture. The most common doubling method applies colchicine to the plant. However, colchicine is phytotoxic and can induce a high frequency of plant death. In this experiment, anthers from two wheat genotypes (“Pavon 76” and ‘Centurk’) were placed on nine embryoid initiation media having three sugar sources (maltose, sucrose, and maltose + glucose) with three colchicine concentrations (0.0, 0.1, and 0.2 g · l^-1). Wheat starch was used as a gelling agent. After three days, the anthers were washed and moved to fresh media without colchicine. Increasing the colchicine concentration decreased the number of embryoids produced from 77.4 embryoids/100 anthers to 29.9 embryoids/100 anthers, but did not significantly affect the frequency of plant regeneration (0.49 green plants/embryoid to 0.40 green plants/embryoid), and increased the frequency of doubled-haploid plants (19.0 doubled-haploid plants/100 green plants to 72.3 doubled-haploid plants/100 green plants). Considering the total number of doubled-haploid plants produced, low levels of colchicine added to the initiation media were very effective.     

Aneuploid Analysis of Anther Culture Response in Wheat

Genetic stocks of Triticum aestivum including the disomic, 8 ditelosomic and 3 nullisomic-tetrasomic ‘Chinese Spring’ wheat lines were employed to ascertain the chromosomal arm locations of the genes acting on the production of embryos from micro-spores and on the regeneration capacity (green and albina) of the microspore-derived embryos. All these aneuploid lines differed significantly from the parental line ‘Chinese Spring’ for embryo production. Our results confirmed or in most cases established that genes affecting embryo production are located on several chromosomal arms: IBS, 1BL, 3AS, 3AL, 5AS, SAL, 5BS, 5BL, 7DS, 7DL. Whereas most of the chromosomal arms stimulate the production of embryos from the microspores, IBS and 1BL reduce it. The results of plant production from microspore-derived embryos suggest that the genes increasing regeneration ability are located on CS5A chromosome and are likely associated to a gene increasing green plant frequency. On the contrary, the 1BL arm increases the albina frequency.     

小麦花药培养的基因型效应及优良基因型筛选

为进一步提高小麦花药培养的效率,从而加快花培育种的进程,对140份不同基因型的小麦亲本及其杂种F1和F3代进行花药培养,并对各世代不同基因型小麦的花药培养力进行研究。结果表明：小麦不同基因型及亲本的花药愈伤组织诱导率在2.89%~41.25%之间,平均为3.55%,绿苗分化率在0%~70.00%之间,平均为33.05%;114份不同基因型小麦F1代杂种材料中,有107份材料能够诱导出愈伤组织,愈伤组织诱导率最低为0%,最高为76.25%,平均为11.59%,绿苗分化率在0%~160.00%之间,平均为29.20%;F3代材料均能诱导出愈伤组织,愈伤诱导率在4.58%~80.83%之间,平均为16.39%,绿苗分化率在0%~71.43%之间,平均为34.99%。因此,不同基因型小麦材料的愈伤组织诱导率、绿苗分化率和绿苗产率在各材料间差异很大。依据高培养力组合亲本组配的规律,筛选到可作为桥梁亲本的材料（品种）有：‘宁春4号’、Jinghong No.2、‘鉴63’、Chuanmai 18、Synthderiv.29589、S111、D-27、‘晋匽746-9’和K6-39等。     

The Effect of Gelling Agents on Wheat Anther and Immature Embryo Culture

This study was conducted to determine whether inexpensive starches and various agars could replace Ficoll, which is very expensive, and Bacto-agar, which may be inhibitory, in wheat (Triticum aestivum L.) anther and immature embryo culture. Anthers from Pavon 76’ had significantly greater embryoid-initiation frequencies (number of embryoids per anther) on PI media with corn (0.67) and wheat starch (1.50) than with Ficoll (0.47). Anthers on 85D12 media with Ficoll, corn starch and wheat starch had similar embryoid-initiation frequencies (0.51, 0.50, and 0.52, respectively). The embryoid-regeneration frequencies (number of regenerating embryoids per embryoid plated) on 85D12 media containing corn (0.32) and wheat (0.36) starch were significantly higher than on 85D12 with Bacto-agar (0.14). Embryoids on MS media with Bacto-agar, corn starch, and wheat starch had similar embryoid-regenerating frequencies (0.15, 0.20, 0.22, respectively), which were greater than with some of the other agars tested (0.05 to 0.17). When wheat starch replaced Bacto-agar in immature embryo cultures, the frequency of responding embryos was unchanged, but the frequency of green plants regenerated increased an average of fourfold for two cultivars.     

The Effect of Sugars and Growth Regulators on Embryoid Formation and Plant Regeneration from Barley Anther Culture

The influence of carbon source and growth regulator composition in induction medium on anther culture response was investigated using spring barley genotypes. Anthers were cultured on BAC3, Ficoll-containing medium, supplemented with one of the following carbohydrates: sucrose, maltose, cellobiosc and melibiose (6 % w/v). The use of either maltose or cellobiose resulted in a significantly higher anther response, calli and/or embryoid production and green plant regeneration compared to the incubation of anthers on a medium containing sucrose. Contrary to these results, the replacement of sucrose by melibiose in BAO medium, drastically reduced the efficiency of anther culture. As an average for the three genotypes tested, the frequency of green plants per 100 anthers plated was 9- to 22-fold higher on medium supplemented with sucrose or cellobiose than on medium containing melibiose as a sole carbohydrate. Among the growth regulators tested, the combination of auxin NAA (2 mg/l) and cytokinin BAP (1 mg/1) performed much better than the employment of auxin 2,4-D combined either with zeatin riboside or BAP as cytokinins. The beneficial effect of medium supplemented with NAA and BAP was associated with better embryoid formation compared to the other growth regulator combinations tested. The hormone-free combination gave a similar anther response and production of calli as any medium supplemented with growth regulators, but the regeneration capacity of calli produced on hormone-free medium was much lower, resulting in the drastic reduction of the number of both total and green regenerants.     

根据花培育种特点谈小麦花培育种的亲本选配

分析了花培育种的特点,并据此提出花培育种的亲本选配的原则,即在参考常规杂交育种亲本选配的一般原则的基础上,选择农艺性状优良品种作为中心亲本,双亲的差异要尽可能的少,不必过于考虑桥梁亲本的利用。     

小麦花培材料的筛选和利用

花培育种是一条快速有效的育种途径，采用两种培养基对20个小麦组合材料进行花培分析，结果表明：小麦花培愈伤组织诱导培养基K优于C17，绿苗产率是C17的3.5倍多。不同基因型材料花药培养差异较大，筛选出一批不同类型的高培养力基因型材料，为有目的组配杂交组合提供依据，提高花培育种效率。     

培养基对小麦单倍体胚成苗率的影响

为筛选出适宜小麦单倍体幼胚生长的培养基,提高小麦单倍体胚成苗率,将小麦×玉米远缘杂交得到的小麦单倍体幼胚接种在8种培养基上,每种培养基上接种2个材料,平均值用于数据分析。8种培养基中幼苗平均产生率分别为：46.07%、45.90%、34.11%、41.15%、37.99%、36.91%、37.70%和52.51%。结果表明1/2MS +1.0 mg/L KT+3.0 mg/L多效唑+100 mg/L肌醇+5%蔗糖+0.6%琼脂,pH 5.8培养的幼胚成苗率最高,白化苗率低,且单倍体苗的长势较好,有利于培育壮苗。以1/2MS为基本培养基,加入微量的多效唑、KT、肌醇以及较高浓度的蔗糖可以有效促进幼胚生长。     

多效唑在小麦花药培养中应用的研究

在小麦花粉愈伤诱导及分化培养基中,加入一种三唑类化介物---多效唑(MET),对出愈率和绿苗分化率影响不明显,但当其浓度高于2mg/L时,会出现负效应。在小麦壮苗培养基中附加MET3 mg/L,萘乙酸(NAA)0.5mg/L,蔗糖8％,不仅可改善春性小麦花粉植株的生长状态,还可提高花粉苗染色体加倍率。若与将愈伤转入分化培养基以前先在含秋水仙碱250mg/L的MS培养基中处理1天这一方法相配合使用,将会出现更好的加倍效果。冬性小麦花粉植株的壮苗培养基中添加MET,可有效地防止花粉苗在越夏过程中老化,增加分蘖数,从而大幅度提高移栽成活率,但同时花粉植株的染色体加倍率有所下降。     

小麦花药愈伤组织耐盐变异体的筛选

在培养基中加入一定浓度NaCl,对小麦F_1花药愈伤组织的耐盐变异体,进行细胞水平上的筛选.采用直接筛选法及逐渐增加盐浓度筛选法,结果表明,后者是提高耐盐变异体频率的有效措施.C_(17)+0.1%NaCl(转移愈伤组织)C_(17)+0.3%NaCl及C_(17)+0.1%NaCl(转移愈伤组织)C_(17)+0.5%NaCl处理的绿苗分化率,较直接筛选法分别提高5.06及15.0倍.在较高(加0.3%NaCl)盐含量的培养基上分化的花粉植株及其后代(H_2、H_3)表现耐盐性较强.获得了在0.4%含盐量土壤上种植能正常或基本正常生长的高耐盐株系3个.     

The Effect of the "D" Genome from Synthetic Wheat Lines in Anther Culture Responses

Synthetic lines of wheat were compared in order to determine the effect of the “D” genome on embryo induction and plant regeneration from anther culture. The experiments were performed using 6 genotypes of wheat in a randomized block design with 4 replications. In all cases except one, addition of the “D” genome from T. tauschii increased the efficiency of embryo formation and green plantlet production. This positive effect of the “D” genome does not exclude the existence of possible interactions between the three genomes (A, B and D) and/or the effect of each genome separately.     

小麦花药培养的基因型差异与杂交组合配制

选用216个不同基因型F1进行花药培养，研究分析不同基因型的花药培养力反应规律，结果表明，不同基因型的花药培养力差异很大，高培养力及低培养基因型各占1／4左右，而中培养力基因型占1／2左右，不花培育种中的桥梁亲本选择问题进行了分析，筛选出具有高培养力和高度，优质基因的花培桥梁亲本。     

Studies of the Genetic Relationship between Anther Culture and Somatic Tissue Culture Abilities in Wheat

A population of thirty-eight doubled haploid lines, developed from the F₁ between two wheat parents differing in anther culture and somatic tissue culture responses, ‘was used to examine the genetical control of responses to these in vitro systems. During anther culture genetic variation between lines was exhibited for frequencies of callus induction., embryo production and embryo regeneration rates. In addition the relative frequencies of green and albino plants was shown to be genotype dependent. However, there was no correlation, between the frequencies of embryo production and the regeneration rate of those embryos suggesting an independent genetic control of these two components. Transgressive segregation for performance was observed for all components indicating that at least two genes are involved in the response of each, and lines for improved performance, combining high ernoryo production rates and good regeneration capacity were identified. No genetic variation for frequencies of callus induction from immature embryos was observed in this cross. However, genetic variation for the regeneration frequencies of plants was observed. Lines with an improved tissue culture response over the two parents were identified. There was no correlation between the performance of lines in anther culture and somatic tissue culture, indicating separate genetical control, and lines with alternative levels of response to the two systems were identified.     

小麦花药培养的基因型差异与亲本选配分析

对104份不同基因型材料进行花药培养,结果表明：F1代愈伤组织诱导率为13.28%,高于F3代的6.02%; 绿苗产率F1代为2.88%,F3代为1.10%,F1相当于F3的2.6倍。同样的培养条件下,不同基因型材料间花药培养力差异很大,愈伤诱导率在 0～111.43% 之间、绿苗产率在 0～49.29% 之间;愈伤诱导率、绿苗分化率与绿苗产率三者之间成正相关关系。同时筛选出了一批如宁春4号等具有高培养力、高产和优质基因的花培桥梁亲本,为有目的配制杂交组合提供依据。     

Tal小麦花药培养出愈和分化的研究

采用Ｔａｌ小麦轮回选择集团４个不同的可育株和Ｔａｌ、矮败小麦多父本复合杂交３个世代的可育株，分别进行花药培养。结果表明：１．Ｔａｌ小麦高世代具有较高的出愈率和绿苗分化率，但与常规品种间杂交一样，出愈率和绿苗分化率有随世代提高而降低的趋势。２．不同交配方式花培效果有明显差异，其中以轮选群体的出愈率最高，其次是多父本复合杂交后代；回交转育稳定的Ｔａｌ小麦的单交出愈率最低。３．基因型对出愈率、绿苗分化率     

培养基和基因型对粳灿稻杂交种花培的影响

采用三种培养基对不同的粳籼稻杂交种F_1进行花药培养。不同的培养基表现出不同的花培效果,基因型之间的培养力差异显著。培养基和基因型之间存在着一定的互作效应。选择培养力高的基因型和与之相适应的培养基是粳籼稻杂交种花培育种的重要因素。     

基因型在棉花花药培养中的效应

棉花花药培养愈伤组织的诱导是植株再生的第一步，基因型在愈伤组织诱导上的差别必将影响着植株再生。本试验中，亚洲棉愈伤组织诱导率最高，陆地棉次之，海岛棉较差，野生种最差。在花药愈伤组织诱导上，种间杂种没显示出优势，但陆地棉品种间杂交则表现出优势。不同基因型诱导产生的花药愈伤组织的颜色、质地、生长量和生活力不同，亚洲棉生长量最大，但活力较差。     

小麦花培材料适宜接种密度研究

为了使小麦花培育种顺利开展,提高优质小麦花培育种的成功率,摸索了各试验材料适宜的接种密度。采用8种不同优质麦材料（品种、组合）在相同体积培养基里进行接种试验。结果表明,各试验材料适宜的接种密度不尽相同,其中‘石4185’ 、‘周麦16号’ 和‘F129号’最佳的接种密度分别为100~120枚/瓶,90~115枚/瓶,74~115枚/瓶。各试验材料愈伤组织诱导率从高到低依次为‘石4185’、‘周麦16号’、‘F129号’、‘邯6172’、‘F1166号’、‘F1167号’、‘F123号’、‘藁优8901’。研究表明,除了‘邯6172’,‘石4185’、‘周麦16号’和‘F129号’这3个材料与其他材料的愈伤组织诱导率有显著差异可以用作花培研究而其他材料则不建议做深入的花培研究。