玉米单倍体加倍(DH)系籽粒性状表现的研究

玉米单倍体育种是快速选育纯合自交系的方法.研究利用单倍体诱导系,对来自BSSS杂种优势群的两个自交系BX08和BX19的杂交F1进行诱导获得单倍体,又经加倍获得33个加倍纯合系(DH),分析单倍体加倍纯合系与两个亲本及F1杂交种的果穗及籽粒性状表现.结果表明,单倍体加倍系的籽粒容积和质量比亲本籽粒都有所提高,子代的农艺性状获得显著改善,说明发生单倍体不是基因随机分离的结果,单倍体育种存在选择压力,有利于优良基因的累积,淘汰不良基因.     

东北早熟玉米育种方法改良的目标、必要性及主要内容

1改良目标围绕东北玉米种业发展对种质改良及技术需求,重点明确玉米产量及抗性性状基因的调控机制,创新高配合力多抗玉米自交系设计技术;规模化开发育种性状的功能分子标记,实现分子标记辅助育种技术实用化;开发高诱导率的单倍体诱导系和加倍技术,实现双单倍体（DR）育种技术的规模化应用;针对东北极早熟玉米区育种性状改良需求,引进多样性种质,通过表型和基因型评价,发掘优异基因资源,创建有效的种质改良与创新平台。     

同一基础材料的玉米双单倍体(DH)系配合力的分析

利用高频玉米单倍体诱导系,对选系材料M35/F35的F1代进行诱导获得大量单倍体籽粒,经自然加倍获得一批纯合双单倍体(DH)系,用测验系京24组配杂交组合分析DH系的配合力表现。结果表明,来源于同一基础材料的不同DH系之间,配合力差异较大,部分DH系的配合力比亲本增加,说明采用单倍体育种可以选育到高配合力的优良玉米自交系,组配出优良品种,从而加快育种进程,应用前景广阔。     

不同来源的糯玉米单倍体诱导研究

以6个不同来源糯玉米基础材料(人工合成群体、地方品种、单交种)为母本,引进的3个单倍体诱导系为父本进行杂交诱导,对糯玉米单倍体诱导率及化学加倍效率进行研究。结果表明,单倍体诱导率受母本基础材料和父本诱导系影响均较大,而散粉株率、结实株率则主要受母本基础材料的影响。从基础材料的来源看,单倍体诱导率从高到低依次是人工合成群体、地方品种、单交种,散粉株率和结实株率从高到低依次是地方品种、人工合成群体、单交种。从诱导系的单倍体诱导率来看,YD 2的诱导率为5.3%,显著高于YD 1和YD 3,在糯玉米单倍体诱导育种中可加大对YD 2的利用。     

玉米DH系的表现及其同工酶电泳分析

以孤雌生殖诱导系高诱1号诱导的单倍体植株为材料,由单倍体加倍后获得的DH系性状上表现高度一致,其中一些DH系的长势和果穗性状达到了育种上的要求.DH系的过氧化物酶和酯酶同工酶电泳表明,部分DH系具有母本谱带,部分具有父本谱带,由此说明,孤雌生殖诱导系诱导单倍体具有一定的随机性.     

孤雌生殖诱导玉米单倍体育种技术研究

玉米单倍体育种技术可以有效地缩短育种进程,快速获得大量纯合系。孤雌生殖诱导是获得玉米单倍体的主要方法之一,该项技术在国外各大育种公司已被广泛使用,但在国内仍处于研究探索阶段。本文从诱导系的选择与繁殖、基础材料的组配、杂交诱导、单倍体籽粒的挑选、单倍体植株的种植、加倍以及双单系的繁殖等方面综述了孤雌生殖诱导系（简称诱导系）产生玉米单倍体育种技术的基本流程,同时展望了单倍体育种的应用前景。     

玉米单倍体DH系特点及应用研究进展

利用单倍体诱导系选育技术能显著加快育种进程,高效单倍体诱导材料的培育及单倍体DH系应用是玉米单倍体育种的重要基础.本文就玉米单倍体DH系的特性及应用加以阐述,并对今后玉米单倍体DH系研究进行了展望.     

同一来源玉米DH系的性状分离分析

利用单倍体诱导系对遗传背景相近的玉米自交系MC1和MC2所组配的F1进行杂交诱导获得单倍体,经过自然加倍得181份纯系(DH系),分析DH系农艺性状的分离情况。结果表明:181个DH系株高、穗位高、穗长、秃尖长、穗行数、百粒重和单穗粒重等数量性状的次数分布符合正态分布;所得DH系各数量性状的变异类型丰富,且在产量性状上存在优于两亲本的分离;两亲本的21对SSR核心引物分子标记带谱在DH系中的分布都近似于1∶1比率,未出现偏分离。因此,通过遗传诱导的方法所得DH系类型多样,可以满足育种上的需要。单倍体育种是一种选育优良玉米纯系的有效方法。     

CENH3介导的单倍体诱导技术研究进展

传统育种方法选育纯合自交系需要7~9个世代,含单个亲本染色体的单倍体经过染色体加倍一个世代就可以获得双单倍体(doubled haploid,DH),加快育种进程。许多单倍体获得和加倍的方法受物种或基因型限制,不能广泛应用。在拟南芥中通过遗传工程操控着丝粒特异组蛋白CENH3(centromere-specific histone 3 variant)可产生单倍体。CENH3是细胞分裂时染色体分离所必需的,其决定着丝点的定位。序列替换或点突变的CENH3以及异源CENH3可补偿cenh3的突变,形成单倍体诱导系,同野生型植株杂交诱导单倍体的产生。本研究论述了CENH3介导的单倍体诱导技术的研究进展。CENH3在植物中广泛存在,CENH3介导染色体消失诱导单倍体技术具有广阔的应用前景。     

优良玉米单倍体诱导系丹诱3号的选育与应用

玉米单倍体诱导系丹诱3号是丹东农业科学院以Stock 6×T 68为基础材料,再复合杂交W 23ig和高诱1号,利用单倍体技术选育的优良诱导系,整合优势诱导材料,实现优良基因的累加聚合。丹诱3号标记清晰,对环境反应钝化,在不同生态区域保持较高诱导率,特别是对含有旅大红骨种质资源的材料诱导率较高。截至目前,利用其诱导加倍选系组配的玉米杂交种有5个通过审定,具有良好的应用前景。     

Properties of maternal haploid maize plants and potential application to maize breeding

Summary Presented are the results of a two-year study of haploid maize plants in the field. The haploids were produced with the aid of inducer line ZMS. In total, 604 and 1030 haploids were obtained and studied in the first and second years, respectively. Tassels of haploid plants were found to be almost completley sterile. Fertility of ears was studied by pollinating them with the pollen from diploid inbred lines, the cross resulting in almost all of the haploid ears carrying kernels. On average 27.4 kernels per ear of haploid plant were obtained in the first year of study and 26.3 in the second. These gave rise to normal diploid plants. This property allows genotypes selected at the level of haploid plants to be involved in breeding process. Unusual plants were found among haploids, phenotypically resembling homozygous lines. It was assumed that the plants had resulted from spontaneous chromosome doubling in haploids. The results of comparative studies of progenies of unusual plants and inbred lines derived from the same synthetic population are presented.     

利用高油分的花粉直感效应鉴别玉米单倍体

通过对农大高诱1号衍生材料的诱导率和含油量的分析,筛选出了一个诱导率与含油量均较高的高诱株系.该系油分具有显著的花粉直感效应,父本效应值为0.38,以其为父本与普通玉米杂交,杂交当代籽粒含油量显著高于自交籽粒和单倍体籽粒,花粉直感的增油率在30%以上.利用这一区别所发展的单倍体油分筛选法准确率超过90%,明显高于标记     

小麦与玉米杂交产生小麦单倍体与双单倍体的稳定性

小麦与玉米杂交是诱导小麦单倍体最有效的途径之一, 但单倍体和双单倍体产生频率不稳定影响了该技术的应用。选用13个小麦杂种F₁代单交组合与玉米杂交, 研究了不同小麦生长环境、生长素处理、培养基和壮苗处理对单倍体及双单倍体产生频率的影响。小麦生长在大田, 去雄后割穗培养与玉米杂交平均得胚率为23.9%, 每个杂交穗平均得胚数6.8个, 均是返青后从大田移回冷温室盆栽的3倍以上;不同小麦杂交组合间胚产生频率存在明显差异。生长素Dicamba蘸穗处理平均得胚率是21.5%, 与2,4-D处理得胚率(21.1%)无显著差异, 但不同杂交组合间差异显著。B5培养基幼胚萌发率为70.9%~88.3%, 平均82.0%;1/2 MS培养基胚萌发率为70.0%~86.0%, 平均76.6%;两种培养基平均胚萌发率无显著差异。试管苗经壮苗培养基壮苗处理与试管苗经移栽壮苗处理后加倍效率分别是67.6%和8.6%。移栽壮苗处理的苗分蘖少, 生长较弱, 加倍处理后存活率低和加倍率低是其单倍体加倍效率低的原因。     

玉米高频率孤雌生殖单倍体诱导系的选育与鉴定

本试验从单倍体诱导系Stock6与高油玉米群体BHO的杂交后代中经过不断的测交和选择, 育成了我国第一个孤雌生殖单倍体诱导系农大高诱1号。 在16个测交组合中高诱一号对411自交系的单倍体诱导率最高(9.2%); 而对H4自交系的单倍体诱导率最低(1.92%); 其平均单倍体诱导率为5.34%。 以黄绿苗自交系Syn695yg为测验种, 高诱1号的     

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。     

Genetic Variability for Haploid Production in Crosses Between Tetraploid and Hexaploid Wheats with Maize

Crosses were made between 14 wheat genotypes (11 tetraploid, 3 hexaploid) and a single Fl hybrid of maize that was used as the male parent. The experimental design consisted of randomized blocks with three replications. Plants were grown under controlled greenhouse conditions (day length 16 h and temperature 25 °C/15 °C, day/night). To enhance embryo survival, 2, 4-D treatment (10 mg/1) was applied to spikes 24 h after pollination with maize. Embryos were recovered from all tetraploid and hexaploid wheats at a rate of 2.09 to 26.76 per 100 pollinated florets. Haploid and doubled haploid plants were obtained from all hexaploid genotypes (T. aestivum) and from 5 of 11 tetraploid genotypes (T. turgidum var.). The most important point of these experiments was the ability to produce haploid plants from tetraploid wheat for two reasons: firstly, anther culture cannot be applied in tetraploid wheat (T. turgidum var.) due to the inefficiency of embryo formation and the high proportion of albino plants. Secondly, to date, crosses between tetraploid wheat and maize have resulted in embryo formation, but not in haploid plants.     

Improved Techniques for Haploid Production in Wheat using Chromosome Elimination

Wheat × maize and wheat × pearl millet crosses have proved efficient for haploid production using various genotypes of wheat; 22 and 27 % of florets produced embryos. In favourable conditions 6—9 haploid plants per spike were produced. The following simplifications or improvements in technique are recommended: 1. Only a single treatment with an aqueous solution of dicamba or 2,4-D (50–100 ppm) for embryo stimulation in vivo; 2. Application by spraying or dipping the spikes; 3. Application time two to four days after pollination; 4. Embryo rescue 15 to 18 days after pollination; 5. Crosses without emasculation are possible if pollination occurs 1–2 days before anthesis. More than 450 haploids and some doubled haploid (DH) lines (after colchicine treatment in vitro) were produced using these methods. No hybrid plants, chromosome additions or substitutions were found.     

Molecular identification of Pm12-carrying introgression lines in wheat using genomic and EST-SSR markers

The traditional process of obtaining maize hybrids involves the generation of inbred lines through successive generations of selfing and subsequent testcrosses in order to identify the best combining ability by allelic complementation. A fast alternative to obtain inbred lines is to induce the formation of haploids followed by chromosome doubling. However, even with the aid of haploid-inducing genetic sources, this strategy has not been widely used in maize breeding programs, partly due to difficulties inherent to haploid generation and identification. In order to evaluate the possibility of using dihaploids to generate homozygous maize tropical lines, we used the androgenetic haploid inducer line W23 as a female parent in crosses with the tropical single-cross hybrid BRS1010. Within the progeny of these crosses, 462 seeds were phenotypically selected as putative haploids by the purple-colored endosperm and colorless embryo conditioned by the R1-nj gene. Among these, only four individuals were confirmed as being haploids using SSR markers, chromosome counting and flow cytometry, showing that the phenotypic marker was not efficient in detecting haploids in the tropical maize genotype used. All four haploids as well as some diploid plants presented reduced size, corroborating the difficulties for haploid identification by phenotypic evaluation. Genetic diversity analysis revealed by SSR markers divided the haploids in two groups represented by flint and dent maize inbred lines, which could be helpful in identifying complementary dihaploid lines. The present article demonstrates that a combination of haploid production and SSR fingerprinting is a feasible strategy for maize hybrid development in tropical germplasm.     

玉米单倍体雌雄育性的自然恢复以及染色体的化学加倍

本试验利用孤雌生殖诱导系所创造的单倍体为材料, 对单倍体雌雄育性的自动恢复 现 象以及染色体的化学加倍方法进行了初步研究。 结果表明, 平均有93.2%的单倍体雌穗 是自 然可育的, 每穗结实粒数平均为25粒。 相比之下, 雄穗的自然育性大大偏低, 但从不同 基因 型中诱导出来的单倍体的雄穗自然育性存在巨大差异, 其中     

Efficient Production of Doubled Haploid Plants through Chromosome Doubling of Isolated Microspores in Brassica napus

Three methods of chromosome doubling to produce doubled haploid plants from microspore cultures of Brassica napus were compared: colchicine treatment of microspore-derived plants, microspore-derived embryos, and isolated microspores. In the whole plant treatment, 53% of the treated plants set seed, but the treatment delayed plant growth and reduced seed set. When microspore-derived embryos were treated with colchicine, the doubling frequency was 32% (compared to 15% for spontaneous doubling). Direct colchicine treatment of isolated microspores resulted in a doubling efficiency of 70 % of the whole plants. This treatment also stimulated embryogenesis in microspore culture, leading to increased plant regeneration. Thus, direct chromosome doubling of isolated microspores is efficient and more than 10 000 doubled haploid plants have been produced in this manner in the past three years in order to accelerate the plant-breeding process.