Haploid Wheat Induction by Crosses Between Different Ploidy Wheat and Different Populations of Maize

Haploid embryo and plant producing frequencies were studied by crossesing diploid, tetraploid and hexaploid wheat with landraces,hybrids and inbred lines of maize. It showed significant differences among the wheat and maize populations. The tetraploid and hexaploid wheat were better than the diploid. High frequencies were obtained by using tetraploid wheat of Triticum turgidum cv. TG14 and maize landraces cv. Xiaoyumi and Xiao Huangmaya. The highest haploid plant producing frequency (6.95 %) was obtained in the TG14 × Xiaoyumi.     

Fertilization and Embryo Development in Hybridization Between Wheat and Leymus

Observations were made of the fertilization and embryo development in intergeneric cross between Triticum aestivum L. cv. Chinese Spring and Leyrnus secalinus Tzrel. The pollen germination of Leymus secalinus appeared normal on the stigma of Triticum aestivum and the pollen tubes grew into the style and entered the embryo sacs. Double and simple fertilization were observed in the pollinated florets. Of the 319 ovaries examed 62 (19.44%) had double fertilization and had embryo and endosperm, but endosperm development was slower than that of the embryos, 49 (15.36%) had only embryo and 7 (2.19%) had only endosperm. The total percentage of fertilization was as high as 36.99%. However, only I seed was obtained from 150 wheat ftorets pollinated with Leymus secalinus. This was obviously due to the absence or poor development of the endosperm. It may be suggested that the potential of increasing the frequency of hybrid plant obtainment was great in the cross between wheat and leymus, if embryo culture technique is employed at the early stage of hybrid embryo development.     

The Production of Haploid Wheat Plants via Wheat X Maize Hybridization

The intergeneric cross between wheat and maize are characterized by a high frequency of formation of hybrid embryos, but maize chromosomes are rapidly eliminated in the first few cell division cycles to produce haploid wheat embryos. If left the embryos on the plants they will soon abort, as a result of the absence or poor development of the endosperm. Embryo rescue techniques should not enable these embryos to grow to plants because of the embryos were extremely young for embryo culture. Viable embryos were obtained at much higher frequenc, if spikes containing cross-pollinated florets were dipped in 100 ppm 2,4-D solution 4 hours after pollination. Of the 382 florets treated, 64(16.8%) embryos were obtained 10 days after treatment, and 47 plants recovered on the culture medium. In control (2,4-D not applied) only 1 (0.96%) embryo and 1 plant was obtained from 104 florets. This simplified technique should enable haploid wheat plants production through wheat - maize to apply to practical breeding.     

小麦×高粱的受精率和小麦单倍体的诱导(简报)

远缘杂交无论在植物遗传学理论的建立还是在新品种的培育上都是一项十分有用的技术,特别在小麦上成绩更为突出。近来发现小麦不但和其亲缘属(如山羊草属、偃麦草属,赖草属等)植物可以杂交,而且和亲缘关系较远的植物种属如玉米、珍珠栗、大刍草、摩擦禾等杂交也有很高的成胚率,并且通过杂种胚发育过程中父本染色体的消失,可以获得小麦单倍体。Laurie和Bennett以高粱属的     

小麦Kr基因在小麦与玉米或鸭茅状摩擦禾杂交中的失活

用３７个小麦（Ｔｒｉｔｉｃｕｍａｅｓｔｉｖｕｍ）品种（系）为母本,分别与黑麦（Ｓｅｃａｌｅｃｅｒｅａｌｅ）、球茎大麦（Ｈｏｒｄｅｕｍｂｕｌｂｏｓｕｍ）、玉米（Ｚｅａｍａｙｓ）和鸭茅状摩擦禾（Ｔｒｉｐｓａｃｕｍｄａｃｔｙｌｏｉｄｅｓ）杂交,比较其亲和性,小麦和玉米或鸭茅状摩擦禾杂交比小麦与黑麦或球茎大麦杂交的亲和性显著提高。携带着显性Ｋｒ１和Ｋｒ２基因的小麦品种Ｈｏｐｅ与黑麦杂交,不能形成胚,而与玉米及鸭茅状摩擦禾杂交时,成胚率分别达１６．００％和３２．５０％。表明控制小麦与黑麦及球茎大麦杂交亲和性的Ｋｒ基因系统在小麦与玉米及小麦与鸭茅状摩擦禾属间杂交中失活。讨论了还存在有其它控制小麦属间杂交亲和性的遗传调控系统的可能性。     

The effect of the crossability loci Kr1 and Kr2 on fertilization frequency in hexaploid wheat x maize crosses

Summary Dominant alleles of the Kr1 and Kr2 genes reduce the crossability of hexaploid wheat with many alien species, including rye and Hordeum bulbosum, with Kr1 having the greater effect. However, a cytological study of wheat ovaries fixed 48 h after pollination showed that the wheat genotypes Highbury (kr1, Kr2) and Chinese Spring (Hope 5B) (kr1, kr2) were crossable with Seneca 60 maize, fertilization occurring in 14.4 and 30.7% of embryo sacs respectively. The latter figure was similar to the 29.7% fertilization found in Chinese Spring (kr1, kr2). Most embryo sacs in which fertilization occurred contained an embryo but lacked an endosperm and where an endosperm was formed it was usually highly aberrant. All three wheat x maize combinations were karyotypically unstable and rapidly eliminated maize chromosomes to produce haploid wheat embryos.     

不同倍性小麦和玉米不同群体杂交诱导小麦单倍体的研究

自从Ｚｅｎｋｔｅｌｅｒ等［１］首先报道了小麦×玉米受精现象以来,许多学者对六倍体普通小麦与玉米杂交进行了广泛的研究,获得单倍体的普通小麦,并筛选到一些杂交亲和性较高的亲本材料［２］。但四倍体小麦与玉米杂交研究报道较少。ＯＤｏｎｏｕｇｈｕｅ等用四倍体小麦与玉米杂交获得单倍体胚［３］。随后,Ａｍｒａｎｉ等［４］和孙敬三等［５］利用这一方法相继获得四倍体小麦的单倍体苗。本文报道了不同倍性的小麦基因型与玉米不同群体杂交对诱导小麦单倍体的影响１　材料和方法１．１　亲本材料用作母本的二倍体小麦有一粒小麦（…     

Induction of Haploid Durum Wheat Plants Through Pollination of Maize Pollen

When tetraploid wheat (Triticum durum Desf. ) variety DR147 was crossed with maize (Zea mays L. ) variety suppersweet ss 7700, pollen readily germinated on the stigma and one or more pollen tubes reached the embryo sac in 83.4% of wheat florets. The frequency of fertilization and embryo formation was 44.5% and 42. 6% respectively. The hybrids were karyotypically unstable and the maize chromosomes were eliminated early in the development. Thus haploid wheat embryos were form. Although the double fertilization frequency of durum wheat X maize was high (32.7%) to form embryos and endosperms, yet the endosperms were highly abnormal. It was very difficult to produce viable mature seeds from the mother durum wheat plants. The survival of hybrid embryos produced by durum wheat X maize could be improved or prolonged by treatment with 100 ppm 2, 4-D (either by dipping inflorescences in solution or injecting 0.3 to 0.5 mL 2, 4-D solution into the uppermost internodes of the wheat stem). 9 to 13 days after pollination, caryopsis were excised from the pollinated spikes and surface sterilized for peeling of the embryos in different developing stages. The embryos were plated on MS solid medium containing 3% sucrose, 200 mg/L casein hydrolysate for embryo rescue. The experimental results revealed that the well developed embryos (larger than 0. 5 mm with scutellum structure) were easy to produce calli by callus induction or produce haploid wheat plants by embryo rescue, whereas the poorly developed embryos (globular, pear or torpedo-shaped embryos smaller than 0.3 mm) responsed very poorly. The germination frequencies of well and poorly developed embryos were 83.3 % and 12.5 %, respectively. Chromosome counts of root tip cells of the rescued plants proved their haploid nature (2n= 2x= 14).     

由小麦×玉米获得的普通小麦加倍单倍体后代的RFLP变异

用小麦（Ｔｒｉｔｉｃｕｍ　ａｅｓｔｉｖｕｍＬ．）的ｒＤＮＡ克隆ｐＴａ７１和与小麦基因组有部分同源性的玉米（ＺｅａｍａｙｓＬ．）ＤＮＡ克隆作探针,对由小麦ｘ玉米获得的普通小麦加倍单倍体（ＤＨ）后代群体进行ＲＦＬＰ分析。结果发现,不但用ｐＴａ７１在这些ＤＨ后代中检测到ｒＤＮＡ所发生的明显的减少和扩增及非转录间隔区的限制性片段长度的变化,而且用与小麦基因组部分同源的玉米克隆ＭＲ１３和ＭＲ５０在一些ＤＨ后代中检测到缺失变异,特别是用ＭＲ１３在普通小麦ＤＨ系的１８号株的基因组中检测到大幅度的限制性片段长度的变化,即原来的４．３ｋｂ的强信号带消失,取而代之的是４０．０ｋｂ、２．５ｋｂ和２．０ｋｂ三条杂交带。这可能与小麦基因组ＤＮＡ较大的重排事件有关,也可能是由外源的玉米ＤＮＡ插入造成的。     

The Importance of Light Intensity for Pollen Tube Growth and Embryo Survival in Wheat x Maize Crosses

The success of Triticum aestivumxZea mays crosses, used to producewheat doubled haploids, is influenced by light intensity. Toexamine the basis for this response, pollen tube growth, embryosurvival and indicators of photosynthetic rate were measuredin two wheat cultivars (‘Karamu’ and ‘Kotuku’)crossed with maize at two irradiance levels (250 or 750 µmolm^-2s^-1, PAR). Pollen tube growth was significantly affectedby light intensity in ‘Karamu’ plants but not in‘Kotuku’ plants, despite both cultivars being pollinatedby the same maize source. The percentage of pollen tubes reachingthe cavity between the ovarian wall and integuments, or in themicropyle of ‘Karamu’ plants at high light intensity(65%) was nearly three-times greater than that at low lightintensity (22%). Thus, either low light intensity can affectthe maternal wheat plant in a way that inhibits pollen tubegrowth and/or high light intensity may promote pollen tube growthin ‘Karamu’ plants. Significant differences in ratesof electron transport in plants grown at the two light intensitiesindicated that the rate of photosynthesis may also have an effecton pollen tube growth. These results have importance for improvingthe efficiency of wheat x maize crosses and other wide cerealcrosses. Copyright 2001 Annals of Botany Company Intergeneric hybridization, light intensity, pollen tube growth, embryo survival, Triticum aestivum, wheat,Zea mays , maize     

RFLP Variations of Common Wheat Doubled Haploid Progenies from Wheat×Maize Crosses

Wheat ( Triticum aestivum L. ) and maize ( Zea mays L. ) crosses (the chromosome elimination system) can be used to produce frequently a large number of doubled haploid (DH) wheat lines by embryo rescue and doubling treatment. The resulting DH lines are genetically homogeneous. Significant RFLP variations were detected in common wheat DH progenies from wheat and maize crosses by using wheat rDNA clone pta71 and two maize DNA clones (MR13 and MRSO) homologous to wheat genome as probes. The results revealed that the copy number and restriction fragment length of rDNA in some wheat DH progenies was changed, and also that deletion was detected in several DH plants when probed with MR13 and MR5O. In particular, the RFLP pattern of DH line No. 18 was greatly changed using MR13 as a probe. In this line, three new bands, 40.0 kb, 2.5 kb and 2.0 kb emerged while a 4.3 kb intense band from the parental common wheat genome disappeared. This change may be related to a quite large DNA rearrangement within the wheat genomic DNA or an insertion by alien maize DNA fragment.     

莜麦与玉米的远缘杂交

莜麦（Ａｖｅｎａ ｎｕｄａ Ｌ．）和玉米（Ｚｅａ ｍ ａｙｓＬ．）杂交有一定的受精率和成胚率。人工传粉后玉米花粉能很快在莜麦柱头上萌发并长入莜麦花柱中,在６８％ 的莜麦花柱中可观察到不只一条玉米花粉管。对授粉后的１６３ 个莜麦子房进行制片观察,发现受精率可达１１．０４％ ,其中只形成胚而无胚乳的卵细胞单受精占３．０７％ ,只形成胚乳而无胚的中央细胞单受精占１．８４％ ,既形成胚又形成胚乳的双受精占６．１３％ 。从授过玉米花粉的大约２２００ 朵莜麦小花中收获１８ 粒种子,对１２ 株实生苗根尖染色体计数表明,其中９ 株为单倍体（２ｎ＝ ２１）,３ 株为二倍体（２ｎ＝ ４２）。结果证明,通过莜麦和玉米杂交后玉米染色体的消除可以获得莜麦单倍体     

Naked Oat×Maize Hybridization

There is a certain frequency of fertilization and embryo productivity in naked oat (Avena nuda L. ) × maize (Zea mays L. ) crosses. The maize pollen readily germinated on the naked oat stigma and more than one pollen tubes grew into the style in about 68% of florets. In a sample of 163 florets fixed after pollination, 5 (3.07%) had only an embryo, 3 (1.84%) had only an endosperm and 10 (6.13%) had both. Overall, 9 haploid and 3 diploid naked oat plants were obtained from 12 seeds which formed following application of maize pollen to about 2200 emasculated naked oat florets. Preliminary studies indicated that elimination of the maize chromosomes occurred early in the embryo and endosperm development. This method gives a new approach for obtaining haplo!d naked oat.     

不同倍性香石竹杂交受精过程及胚胎发育研究

采用石蜡切片法对以四倍体香石竹品种‘紫蝴蝶’(2n=4x=60)为母本,单瓣中间材料‘NH6’(2n=2x=30)为父本杂交后受精过程及胚胎发育进行研究。结果表明:(1)授粉后17h,花粉管进入助细胞并释放内容物,精核进入极核细胞内,与二极核细胞融合形成初生胚乳核;授粉后1d,精核向卵核方向移动,贴伏于卵核核膜上;授粉后2d,形成合子及游离的胚乳核;随后,胚发育经过原胚、球形胚、心形胚、鱼雷形胚阶段。(2)杂交障碍发生在受精过程及胚胎发育的各个时期,表现为:精子与卵细胞不相融合或精子与二极核不相融合、合子未分裂或初生胚乳核未分裂及胚胎的败育。(3)胚败育虽能发生在原胚、球形胚、棒状形胚、三角形胚、心形胚、鱼雷形胚及子叶形胚阶段,但主要发生在球形胚阶段。     

High frequncies of fertilization and embryo formation in hexaploid wheat x Tripsacum dactyloides crosses

The Hexaploid wheat variety Fukuho was crossed with Tripsacum dactyloides (2n=4x=72). The total fertilization frequencies for the egg cell, polar nuclei, and both, were 58.3%, 26.8% and 58.9% of the 168 ovaries examined. However, the fertilization frequency of single polar nuclei was much lower at only 0.6%. The total frequency of fertilization was higher than that in wheat x maize crosses. A total of 49 hexaploid wheat varieties, including Hope carrying the dominant genes Kr1 and Kr2, were crossed with T. dactyloides, and most gave embryos. The embryoformation frequencies ranged from 0.5% to 59.0%. A higher frequency of 32.0% embryo formation was obtained following pollination of the variety Hope. In comparison with embryo formation in wheat x maize crosses the difference of embryo-formation frequencies between the two crosses was significant. The results of high frequencies of fertilization and embryo formation in wheat x T. dactyloides crosses indicated that the Kr genes are as inactive in wheat x T. dactyloides, as they are in wheat x maize crosses, and also that the efficiency of fertilization and embryo formation is higher in wheat x T. dactyloides than in what x maize crosses. The potential of wheat x T. dactyloides crosses for wheat haploid production and wheat improvement is discussed.     

通过和玉米杂交诱导硬粒小麦单倍体

硬粒小麦（Ｔｒｉｔｉｃｕｍ ｄｕｒｕｍ Ｄｅｓｆ．）ＤＲ１４７授以超甜玉米（Ｚｅａ ｍ ａｙｓ Ｌ．） ｓｓ ７７００的花粉后,在８３．４％ 的柱头上观察到花粉萌发及花粉管长入胚囊,有９．９％ 的子房发生了卵细胞的单受精,１．９％ 的子房发生了中央细胞的单受精,３２．７％ 的子房发生了双受精。尽管双受精后可同时形成胚和胚乳,但胚乳往往发育迟缓,甚至败育。硬粒小麦×玉米形成的杂合子核型高度不稳定,在最初的几次细胞分裂中,来自父本玉米的染色体逐步被排除,最后形成硬粒小麦单倍体胚。在授以玉米花粉４ ｈ 后用１００ ｐｐｍ ２,４－Ｄ溶液浸蘸硬粒小麦穗部,可以有效地促进幼胚在缺乏胚乳或胚乳败育情况下的生长和发育。授粉９—１３ ｄ 后由５３３个硬粒小麦子房解剖出２５个胚,得胚率为４．７％ 。通过幼胚拯救获得１１棵正常植株,植株获得率为２．１％ 。根尖细胞染色体计数表明,它们为单倍体（２ｎ＝ ２ｘ＝ １４）。     

Pseudogamous Apomixis in Maize and Sorghum in Diploid-Tetraploid Crosses

Apomictic seed development is a complex process including formation of unreduced embryo sac, parthenogenetic embryo development from the egg cell, and endosperm formation either autonomously, or due to fertilization of polar nuclei by the sperm (under pseudogamous form of apomixis). In the latter case, an obstacle to the normal endosperm development is disturbance of maternal (m) -to-paternal (p) genomic ratio 2m: 1p that occurs in the cases of pollination of unreduced embryo sac with haploid sperms. Usage of tetraploid pollinators can overcome this problem because in such crosses maternal-to-paternal genomic ratio is 4m: 2p that provides formation of kernels with plump endosperm. Using tetraploid lines as pollen parents we observed formation of plump kernels on the ears and panicles of diploid maize and sorghum accessions. These kernels had hybrid endosperm and diploid maternaltype embryo or hybrid embryo with different ploidy level (2n, 3n, 4n). The frequencies of plump kernels on the ear ranged from 0.2-0.3% to 5.7-6.2% counting from the number of ovaries. Maternal-type plants were found in two maize lines, their frequency varying from 10.7 to 37.5% of the progeny plants. In CMS-lines of sorghum pollinated with tetraploid sorghum accessions, the frequency of plump kernels ranged from 0.6 to 14.0% counting from the number of ovaries; the frequency of maternal-type plants varied from 33.0 up to 96.1%. The hybrid nature of endosperm of the kernels that gave rise to maternal-type plants has been proved by marker gene expression and by SDS-electrophoresis of endosperm proteins. These data testify to variable modes of seed formation under diploid × tetraploid crosses in maize and sorghum both by amphi- and by apomixis. Therefore, usage of tetraploid pollinators might be a promising approach for isolation of apomixis in maize and sorghum accessions.     

小麦x长穗偃麦草的受精和早期胚胎发育

用石蜡切片法,对小麦(Triticum aestivum)和长穗偃麦草(Elytrigia elongata)杂交的受精和早期胚胎发育进行了观察。结果表明,长穗偃麦草花粉在小麦柱头上萌发良好,花粉管可顺利长人花柱和胚囊。 观察的170个小麦子房中,17.65％发生了双受精,产生了胚和胚乳;9.41％发生了单卵受精,只产生胚而无胚乳;4.71％。发生了单极核受精,只产生胚乳而无胚;总受精率为31.77％;成胚率为27.06％。由于胚乳的缺乏或发育异常及败育,最终难以获得有生活力的种子。为小麦与长穗偃麦草远缘杂交提供了细胞胚胎学证据。     

禾本科植物染色体消除型远缘杂交的研究进展

植物远缘杂交是作物育种中广泛应用的技术。除了核型稳定的种间杂交可以获得杂种以外,还可以利用核型不稳定的种间杂交后父本染色体消除的现象,通过胚培养和染色体加倍处理获得加倍单倍体（ＤＨ）植株。然而从小麦×玉米杂交获得的ＤＨ后代与其理论上应完全同质的遗传表现却不相符,总有２～５％的ＤＨ植株发生了形态学变异。最近的研究证明,通过小麦×玉米的受精作用,一些玉米特异ＤＮＡ可以被转移到小麦ＤＨ后代的基因组中。     

玉米染色体G—带带型的研究

本文对3个玉米自交系,及其中两个自交系的杂交F_1有丝分裂早中期染色体的G-带带型进行了比较研究。所有的供试材料G-显带的染色体上都具有两种类型的带纹,我们称A型带和B型带。A型带为沿染色体长轴分布,较细的,密切邻近的多重带纹。不同自交系的A型带带型基本相同,杂交F_1的A型带无明显的异型性。非同源染色体间带型各不相同,某些染色体具有易于识别,特征性较强的A型带标记。B型带一般为深染色的大带,位于染色体的近端区。同一自交系每两个同源染色体的B型带可以配对,不同自交系B型带带型互有不同。杂交F_1某些染色体上的B型带带型异型性明显。具异型性的染色体对中一成员的带型与一个亲本相似,另一成员与另一亲本相似。比较对同一细胞先后作G-和C-显带处理的结果表明,B型带和C-带是相同的。