小麦不同杂种世代花药脱分化特性研究

对5个小麦不同组合基因型的F1，F2，F3花药进行培养，结果表明：不同组合基因型在同一杂种世代的花药愈伤组织诱导率、愈伤组织鲜重和绿苗分化率等方面存在较大差异，而同一组合基因型在不同杂种世代的花药愈伤组织诱导率、愈伤组织鲜重和绿苗分化率等方面差异较小。花药愈伤组织诱导率与绿苗分化率具有显著的相关性。以石4185为亲本配制的两个组合基因型，在F1，F2，F3都表现出较高的花药愈伤组织诱导率和绿苗分化率，说明石4185是理想的花培育种桥梁亲本。     

小麦花培育种技术研究

对5个小麦不同组合基因型的F1,F2,F3花药进行培养,结果表明:不同组合基因型在同一杂种世代的花药愈伤组织诱导率、愈伤组织鲜重和绿苗分化率等方面存在较大差异,而同一组合基因型在不同杂种世代的花药愈伤组织诱导率、愈伤组织鲜重和绿苗分化率等方面差异较小。花药愈伤组织诱导率与绿苗分化率具有显著的相关性(r2=0.903)。以石4185为亲本配制的两个组合基因型,在F1,F2,F3都表现出较高的花药愈伤组织诱导率和绿苗分化率,说明石4185是理想的花培育种桥梁亲本。     

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。     

提高冬小麦花药愈伤组织诱导效率的研究

为了进一步提高小麦花药愈伤组织的诱导效率,以12个黄淮麦区冬小麦杂交F1代为供试材料,比较了不同诱导培养基、培养方式和花药接种密度与共培养子房数等因素对花药愈伤组织诱导及分化的影响。结果表明:CHB培养基诱导效果最好,C17次之,而N6和W14效果较差,不同基因型对不同培养基的反应不同,但大多数材料在CHB和C17培养基上反应较好;培养基附加较低浓度的2,4-D(0.5 mg/L)时有利于形成胚状体并在诱导培养基上一步成苗;液体培养时愈伤组织诱导率最高但对愈伤的进一步分化不利,固体和双层培养时的诱导率最低,而半固体培养时不仅有利于愈伤组织诱导而且对其分化较好,是小麦花药培养的较好方式;花药与子房共培养可以显著提高愈伤组织诱导率,花药接种密度较低(2～2.4枚/mL)时较多的子房(20个)可以起到显著的促进作用,花药接种在高密度(6～7.2枚/mL)或中密度时(4～4.8枚/mL)10个或10个以上的子房诱导效果较好。     

不同基因型水稻花药培养效果的比较研究

为了筛选到花药培养效果好的水稻材料,选取常规籼稻、籼粳杂交稻、籼型杂交稻和粳型杂交稻 4 种不同类型的水稻材料进行花药培养,比较它们在花药培养过程中的愈伤组织诱导率和花药培养效果,建立高效花药培养再生体系。结果表明,禾香 1 号和福稻 99 愈伤组织诱导率较高,分别为 3.01% 和 1.55%,可作为水稻花培育种的优良亲本。以玉晶 91 为亲本配制杂交组合并进行花药培养,结果表明玉晶 91 与某些亲本间具有较好的花药培养配合力和杂种优势,在水稻花培育种中具有一定的潜在利用价值。粳型材料中,甬优 1526、组合圣稻 18-15// 春江 041/ 武运粳 24、中浙粳 18/ 鄂香 2 号和香 1 号 /17J42 均不适合作为水稻花药培养育种的材料。甬优 2640 的花药培养愈伤组织诱导率和绿苗分化率均较高,可以在水稻花培育种和DH 群体创建中加以利用。这对创制新的种质资源、构建 DH 群体和培育新品种具有十分重要的意义。     

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。     

基因型对陆地棉花药离体培养反应的影响

对２７种棉花基因型的花药在离体培养中的反应进行了比较,结果表明：所有的基因型在合适的培养条件下均能高频率地形成愈伤组织,但不同基因型间愈伤组织诱导率和诱导量不同;再生植株的花药比原始亲本的花药易诱导出愈伤组织;品种间杂种表现出部分优势,但不明显。本试验中,仅鲁棉６号和Ｓｉｏｋｒａ１－３两个品种诱导获得了胚性愈伤组织、胚状体和再生植株,且二者之间也存在着差别。从基因型在棉花花药培养中的效应、外界调控对棉花不同基因型花药培养效果的影响、棉花花药培养与体细胞培养基因型差异的比较等三个方面进行了讨论,提出了棉花花药培养愈伤组织诱导和植株再生是受两套遗传基因控制的遗传性状,并提出了棉花组织培养植株再生的阈值问题     

小麦花药培养中亲本配合力研究简报

利用双列杂交ＮＣⅡ设计配制了１８个组合进行药培养，研究小麦亲本组合对愈伤组织诱导和绿苗分化的配合力。试验表明，一般配合力高是花培选配亲本的基础，本试验中，愈伤组织，绿苗诱导率和一般配合力均高的亲本是“日本、号”和“８４－２０”。     

甘露醇预处理对不同基因型加工番茄花药愈伤诱导率的影响

采用6个加工番茄F_1为试验材料,研究不同浓度和不同时间甘露醇预处理对形成加工番茄花药愈伤组织的影响。结果表明:利用甘露醇预处理加工番茄花药时,有3个杂交组合在0.2 mol/L甘露醇培养基上诱导4 d时,花药愈伤组织诱导率极显著高于未进行甘露醇预处理的;有2个杂交组合在0.4 mol/L甘露醇培养基上诱导4 d时,花药愈伤组织诱导率极显著高于对照;而杂交组合20040805×P1在0.2 mol/L或0.4 mol/L甘露醇培养基上经过4 d的预处理,花药愈伤组织诱导率均极显著高于对照。说明不同基因型加工番茄花药培养利用甘露醇预处理时,需要采用不同的浓度和处理时间。     

BN对小麦花粉愈伤组织诱导频率的影响

BN对小麦花粉愈伤组织诱导频率的影响结果表明，在W14＋2.0mg/l 2,4-D KT0.5mg/l 11%蔗糖诱导培养基中添加BN可以明显缩短出愈时间，提高花药愈伤组织诱导率。BN在低浓度1.0-1.5μg/l时对愈伤组织诱导起促进作用，提高花药愈伤组织诱导率1．7－4．2个百分点；而当浓度升至10．1μg/l时，则有抑制愈伤组织形成的趋势。     

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。     

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。     

Relative Efficiency of Anther Culture and Chromosome Elimination Techniques for Haploid Induction in Triticale × Wheat and Triticale × Triticale Hybrids

Summary The study was undertaken to evaluate the relative efficiency of anther culture and chromosome elimination (by crosses with maize) techniques of haploid induction in intergenotypic triticale and triticale × wheat hybrids. For this, 15 triticale × wheat and 8 triticale × triticale F₁ hybrids were subjected to anther culture and were also simultaneously crossed with the `Madgran Local' genotype of maize (Zea mays L.) to induce haploids through the chromosome elimination technique. The haploid embryo formation frequency through the chromosome elimination technique was significantly higher in both, triticale × wheat (20.4%) and triticale × triticale (17.0%) F₁ genotypes, as compared to the calli induction frequencies through anther culture (1.6 and 1.4%, respectively). Further, four triticale × wheat and three triticale × triticale F₁ genotypes failed to respond to anther culture, whereas, all the F₁ genotypes formed sufficient number of haploid embryos through the chromosome elimination technique with no recovery of albino plantlets. The haploid plantlet regeneration frequencies were also significantly higher through the latter technique in both triticale × wheat (42.7%) and triticale × triticale (49.4%) F₁s as compared to anther culture (8.2 and 4.0%, respectively), where the efficiency was drastically reduced by several constraints like, high genotypic specificity, low regeneration frequency and albinism. The overall success rates of obtaining doubled haploids per 100 pollinated florets/anthers cultured were also significantly higher through the chromosome elimination technique (1.1% in triticale × wheat and 1.5% in triticale × triticale hybrids), proving it to be a highly efficient and economically more viable technique of haploid induction as compared to anther culture, where the success rates were only 0.2% and 0.1%, respectively.     

Die Nutzung der Antherenkulturmethode im Zuchtprozeß von Winterweizen

Anther culture in the breeding process of winter wheat. III. Ability of winter wheat F₁ populations with the two heterozygous 1AL–IAS/1AL–IRS and 1BL–1BS/1BL–IRS chromosome pairs Application of anther culture to four F₁ hybrids between the IBL–IRS (‘Amigo’) and several 1BL–IRS wheat-rye translocation forms yielded 129 green pollen plants in an average embryo induction frequency of 17.6 %. A total of 2632 anthers was inoculated. 25 % and 42 % of the regenerated plants were haploid and spontaneously doubled haploid, and 33 % had abnormal chromosomal structure. After chromosome doubling treatment 87% of all pollen plants set seeds. By means of multiple peroxidases and Giemsa C-banding patterns, the anther culture progeny could be further classified into 16 plants without the short arm of IR-chromosome of rye, 21 IAL–IRS and 50 1BL–IRS translocation lines and into 16 IAL–IRS, IBL–IRS double translocation lines according to the four possible characteristic types of F₂ gametes of the tested F₁ hybrids. Advantages of the haploid technique for the selection of desirable traits and the meaning of the IRS genes in wheat are discussed.     

Production of Doubled Haploids by Anther Culture and Wheat X Maize Method in a Wheat Breeding Programme

Utilization of the doubled haploid method of breeding usually shortens the time to cultivar release, and methods of haploid production need evaluation in a breeding programme. Thirty-eight different three-way crosses were tested for anther culture response. On average 5.8 percent of the anthers cultured produced calli. Three crosses were found recalcitrant for callus induction. Overall, the anther culture method produced 0.6 plantlet per 100 anthers cultured. Five crosses with an average of 5.8 and 2.8 percent of anthers producing calli and plantlets, respectively, were compared using anther culture and wheat × maize crosses. Non-responsive genotypes for callus induction and plantlet formation in the anther culture method proved to be good parental material in wheat × maize crosses. The average percentages of embryo formation and plantlet production in wheat × maize crosses were 10.3 and 4.7, respectively. Anther-derived plants were cytologically unstable, whereas all the plants regenerated from wheat × maize crosses were haploids (n = 21 chromosomes). The chromosome numbers of the polyhaploids were doubled with a colchicine treatment. Improvement of the two haploid production methods to facilitate their efficient use in a breeding programme is discussed.     

Die Nutzung der Antherenkulturmethode im Zuchtprozeß von Winterweizen

Anther culture in the breeding process of winter wheat. I. Ability of 1B—-1R wheat-rye translocation forms for androgenesis 45 winter wheat varieties or F₁ hybrids, F₂ populations and lines with 1B—1R wheat-rye translocation were tested for their anther culture ability. A total of 48058 anthers was cultured on Potato-2 medium. When averaged over all genotypes and the two experimental years frequencies of embryoid formation of 5.4—6.8 per 100 anthers were observed. Plant regeneration efficiency from embryoids ranged from 5.3—9.1 % or a mean of 4—5 green plants per 1000 anthers plated. The results confirmed the preferential regeneration frequency of gametes with the 1BL—1RS chromosome compared to the gametes with the 1BL—IBS chromosome. Multiple peroxidase were used as marker. The effect of cold pretreatment or of media on the androgenetic response and productivity was not important. On the contrary the variability between the anther response from single ears of the same genotype was noticeable. Examples are presented for the transferability of the androgenetic ability to breeding material. Most green plants obtained were haploid or spontaneous doubled haploid. By cloning it was guaranteed, that progenies were obtained from most of the haploids after colchicine treatment.     

Effect of parental genotypes and colchicine treatment on the androgenic response of wheat F1 hybrids

The effect of the parental genotypes and colchicine treatment on the androgenic response of wheat (Triticum aestivum L.) F1 hybrids was studied. For this, anthers from three F1 hybrids and their parents were cultured on W14 initiation medium and W14 supplemented with 0.03% colchicine. The number of responding anthers, microspore‐derived structures/100 anthers, green plants/embryos cultured, green plants/100 anthers and albino plants/100 anthers were recorded. It was observed that embryo formation and plant regeneration ability were genetically controlled and genotype dependent. In both treatments the variety Kavkaz had a significantly higher percentage of responding anthers, microspore‐derived structures and green plants/100 anthers than the other genotypes. On the other hand, the variety Myconos also demonstrated high microspore‐derived structure production and green plant regeneration when treated with colchicine. The good response observed in these two varieties indicates the importance of colchicine treatment only for certain genotypes. Green plant production capacity of the hybrids was intermediate to that of the parental varieties. As one parent with a high or even an intermediate response to anther culture could lead to the production of sufficient (for breeding purposes) green plants from the F₁ hybrids, it was concluded that screening the inbred lines for the response to anther culture with and without colchicine treatment could contribute to utilization of breeding material with a low response to anther culture via the proper hybrid combinations.     

Anther culture of recalcitrant indica × Basmati rice hybrids

Fertile, green, di-haploid plants were obtained at high frequencies from several indica × Basmati rice F₁ hybrids and/or F₂ plant populations using an improved anther culture procedure. Anthers from cold-pretreated (10 ^°C for 10 d) panicles of six indica (HKR120, HKR86-3, HKR86-217, PR106, Gobind andCH2 double dwarf) and two Basmati rice (Basmati 370,Taraori Basmati) varieties and 14 heterotic indica ×Basmati F₁/F₂ hybrids were cultured in modified agarose-solidified N6M, Heh5M and RZM media. Best callus induction frequencies (2.6–78%) were obtained in RZM medium containing 4% (w/v) maltose,2,4-D, NAA and kinetin. F₂ plants compared to F₁ hybrids and parental rice varieties, were more responsive to anther culture. Androgenesis frequencies of 31–78% were obtained for indica × Basmati F₂ plants in RZM medium in just 30 d which are comparable to or higher than that reported for japonica rice varieties and hybrids involving japonica rice parent(s). Agarose (1.0% w/v)-solidified MS medium containing 3.0% maltose, kinetin, BAP, and NAA, induced green shoot regeneration in 0–51% of the anther-derived callide pending upon the genotype. High plant regeneration frequencies (67–337 green plants per 1000 anthers)were obtained from anther calli of several F₁hybrids (Gobind × Basmati 370 and HKR120 ×Taraori Basmati) and F₂ plants (Gobind × Basmati370, Gobind × Taraori Basmati, HKR86-3 × TaraoriBasmati). A sample of 498 plants obtained from the above hybrids, were transferred to pots with>90% survival; 8–78% of these plants had >5%spikelet fertility and were diploid. In addition,18% of the haploid plants could be diploidized by submerging in 0.1% colchicine solution for 16–18 h. The improved anther culture procedure reported here, resulted in several fold increase in the recovery of green plants from recalcitrant indica × Basmati rice hybrids compared to previous published procedures. The study may accelerate the introgression of desirable genes from indica into Basmati rice using anther culture as a breeding tool. This revised version was published online in July 2006 with corrections to the Cover Date.