杨树三倍体选育研究进展

杨树三倍体具有速生、纤维长等特点,是速生短周期纸浆材的首选树种。本文从发现杨树天然三倍体,人工培育杨树三倍体,特别是在采用杂交、筛选天然2n花粉、人工诱导2n配子方法培育三倍体等方面,对杨树三倍体的育种研究现状进行了综述,指出了杨树三倍体育种的发展方向及意义。     

杨树抗冻性的研究现状

杨树是世界上广泛栽培的重要造林树种之一,我国是世界上杨树资源丰富的国家,杨树具有速生丰产、实用性强、无性繁殖能力强,且基因组较小等特点,现已成为研究林木生理和基因工程研究的模式植物。本文概述了国内外有关杨树抗冻性的生理生化性质及其指标、天然杂种的选择以及人工杂交选育种等方面研究现状,并展望了杨树基因工程在杨树抗冻性遗传改良中的应用前景,此外,对信后如何开展杨树抗冻改良育种的研究提出了一些设想。     

杨树抗冻性的研究现状

杨树是世界上广泛栽培的重要造林树种之一,我国是世界上杨树资源丰富的国家。杨树具有速生丰产、实用性强、无性繁殖能力强,且基因组较小等特点,现已成为研究林木生理和基因工程研究的模式植物。本文概述了国内外有关杨树抗冻性的生理生化性质及其指标、天然杂种的选择以及人工杂交选育种等方面研究现状,并展望了杨树基因工程在杨树抗冻性遗传改良中的应用前景。此外,对今后如何开展杨树抗冻改良育种的研究提出了一些设想。     

三倍体杨树核型分析

对4种三倍体杨树核型进行了比较分析,三倍体毛白杨核型为：2n=3x=57=1M 31m 10sm 15st;三倍体I-214为：2n=3x=57=2M 33m(2SAT) 10sm(1SAT) 10st(3SAT) 2t;三倍体武黑1号为：2n=3x=57=2M 38m(4SAT) 14sm(2SAT) 3st;三倍体廊坊3号为：2n=3x=57=35m 10sm(1SAT) 10st(2SAT) 2t(1SAT)。结果显示三倍体毛白杨核型与三倍体黑杨核型有显著不同,表明它们的基因组结构不同,起源各异。     

鱼类人工多倍体育种的研究进展

鱼类多倍体育种是鱼类育种最热的领域之一。本文概述了鱼类人工多倍体育种研究的进展情况,包括鱼类人工多倍体育种的方法和人工多倍体鱼类可育性等方面。鱼类人工多倍体育种的方法主要有物理、化学和生物学方法,三倍体鱼预期是不育的,四倍体是可育的,但目前两性可育的四倍体的报道还很少。本文还对人工多倍体鱼的生物学意义作了简单的阐述。本文旨在为对今后进一步开展鱼类人工多倍体育种研究提供参考。     

麦类作物多胚的研究

对高原602双胚苗的细胞学进行了研究,有3种类型2n-2n、2n-3n和2n-n,出现频率分别为0．16％、0．02％和0．02％。在2n-2n型里,又发现了形态上的差异,即2n（顶芒）-2n（长芒）。对单倍体育种和三倍体的起源进行了讨论。     

人工三倍体新桑品种嘉陵20号的AFLP分析

利用AFLP分子标记技术, 从DNA水平对人工三倍体新桑品种嘉陵20 号(2n=3x=42) 及其亲本以及与嘉陵20号同亲本的2 个三倍体(2n=3x=42) 桑树材料与10 个二倍体(2n=2x=28) 桑树材料进行了遗传背景分析。获得了它们的指纹图谱、遗传距离及遗传相似系数, 并绘制出了它们的UPGMA聚类图。研究结果发现: 嘉陵20 号的母本7920 (2x;♀) 与父本西庆四号(4x; ♂) 间的相似系数为0 6612。这为今后更具强大杂交优势和多倍体优势的新型人工三倍体新桑品种选育时杂交亲本的选择与组配提供了较有用的参考依据。     

乌鳢三倍体诱导及其生长

采用热休克抑制第二极体排放的方法诱导乌鳢(Channa argus)三倍体,以探索人工诱导乌鳢三倍体的理想热休克条件。采用DNA含量测定法和红细胞核大小鉴定法对获得的鱼苗倍性进行鉴定,同时对普通二倍体乌鳢群体与三倍体群体的生长差异进行了比较研究。结果表明,(1)热休克法适宜诱导条件为,(28±0.5)℃培育水温授精后4 min,在水温42℃条件下持续处理3 min,三倍体诱导率最高,达到87.69%;(2)三倍体与二倍体DNA含量差异极显著(P0.01),其比值为1.50︰1.00;(3)三倍体和二倍体在红细胞核长径、红细胞核体积、红细胞核面积等6项指标上存在极显著差异(P0.01);与二倍体相比,三倍体红细胞体积和核体积分别是其1.69倍和1.60倍;(4)在4月龄与8月龄,三倍体的体长和体重比二倍体稍高,但两者差异不显著(P0.05)。本实验结果为进一步开展乌鳢倍性育种奠定了基础。     

三倍体品种的选育和利用

概述了三倍体生物的两大基本特征(细胞巨大性和不育性)及其产生原因,对有很好经济和社会效益的三倍体杨树、甜菜、无籽西瓜和某些海洋生物选育利用作了简要介绍、展示了三倍体品种选育利用的美好前景。     

橡胶树新品种云研77-2和云研77-4的细胞学鉴定及育种过程

对巴西橡胶树(Hevea brasiliensis)新品种云研77-2和云研77-4及其亲本PR107和GT1的染色体进行了细胞学观察,并介绍了它们的育种历程.结果表明:PR107和GT1的染色体数为2n=2x=36,云研77-2和云研77-4的染色体数目为2n=3x=54,均为三倍体;利用特殊父母本建立杂交授粉园获取种子,设定合理的筛选标准在逆境中进行实生筛选是巴西橡胶树新品种选育的有效途径.     

2n egg formation in Populus × euramericana (Dode) Guinier

Three synthetic triploid seedlings were derived from a cross between maternal parent Populus × euramericana (Dode) Guinier and Populus nigra var. italica (Moench.) Koehne via embryo rescue by treating female floral buds with colchicine at certain developmental stages before and after pollination, and identified by flow cytometric analyses. A total of 21 microsatellite primers were used to infer the ploidy of gametes forming the hybrids and the underlying meiotic mechanism. The results suggest that 2n eggs originated from the maternal poplar, contributing the extra haploid genome in the triploids. In addition, first division restitution (FDR) 2n eggs were produced in the maternal tree according to the marker genotypes of these triploids. Cytological analyses of megasporogenesis revealed that meiosis of the maternal parent was asynchronous. This cytological evidence suggests that 2n eggs result from FDR after treatment with colchicine, as a portion of megaspore mother cells (MMCs) were within the first division at the post-pollination stage. These results aid in understanding the mechanism of 2n egg formation in poplar and make the utilization of 2n eggs in polyploid poplar breeding more effectively.     

Chromosome-level genome assembly of a triploid poplar Populus alba ‘Berolinensis’

Many recent studies have provided significant insights into polyploid breeding, but limited research has been carried out on trees. The genomic information needed to understand growth and response to abiotic stress in polyploidy trees is largely unknown, but has become critical due to the threats to forests imposed by climate change. Populus alba ‘Berolinensis,’ also known “Yinzhong poplar,” is a triploid poplar from northeast China. This hybrid triploid poplar is widely used as a landscape ornamental and in urban forestry owing to its adaptation to adverse environments and faster growth than its parental diploid. It is an artificially synthesized male allotriploid hybrid, with three haploid genomes of P. alba ‘Berolinensis’ originating from different poplar species, so it is attractive for studying polyploidy genomic mechanisms in heterosis. In this study, we focused on the allelic genomic interactions in P. alba ‘Berolinensis,’ and generated a high-quality chromosome-level genome assembly consisting of 19 allelic chromosomes. Its three haploid chromosome sets are polymorphic with an average of 25.73 nucleotide polymorphism sites per kilobase. We found that some stress-related genes such as RD22 and LEA7 exhibited sequence differences between different haploid genomes. The genome assembly has been deposited in our polyploid genome online analysis website TreeGenomes ( https://www.treegenomes.com ). These polyploid genome-related resources will provide a critical foundation for the molecular breeding of P. alba ‘Berolinensis’ and help us uncover the allopolyploidization effects of heterosis and abiotic stress resistance and traits of polyploidy species in the future.     

Genetic identification of 91 poplar cultivars based on SSR markers

Accurate genetic identification and relationship analysis of poplar cultivars is necessary to establish commercial poplar plantations and select suitable breeding strategies. In this study, 91 poplar cultivars belonging to four sections (Aigeiros, Tacamahaca, Populus and Turanga) and inter/intra-sectional hybrids were genotyped using 18 polymorphic simple sequence repeat (SSR) markers. In total, 222 alleles were amplified with an average of 12.3 alleles per marker. The mean polymorphic information content and power of discrimination were 0.706 and 0.813, respectively. Five SSR markers (ORPM_103, ORPM_247, GCPM_1048, GCPM_1255 and LG_X_19) constituted a core fingerprint and were sufficient to identify all the tested cultivars. With some notable exceptions, cultivars of the same species generally clustered together in cluster (UPGMA) and ordination (PCO) analyses. Flow cytometry indicated that 11 poplar cultivars were triploid. Among these, seven had three alleles at some loci, suggesting that SSR markers could indicate the ploidy level to some extent. This study provides useful genetic information for the identification and protection of poplar cultivars in China and offers a guideline for the selection of poplar crossing parents based on ploidy level and genetic relationships.     

三倍体白杨杂种无性系指纹图谱的构建

目的:构建三倍体白杨杂种无性系指纹图谱,鉴定三倍体白杨杂种无性系。方法:分离纯化三倍体白杨杂种DNA模板,采用扩增片段长度多态性(AFLP)分子标记技术构建三倍体白杨杂种无性系指纹图谱。结果:从64对引物组合中筛选出M-CTA/E-CAG、M-CAC/E-CCA、M-ACT/E-CTC和M-CTT/E-CTG等4对多态性较高的引物组合,并应用该引物组合对21个三倍体白杨杂种无性系进行了AFLP分析,构建了21个三倍体白杨杂种无性系指纹图谱。结论:构建无性系指纹图谱是鉴别三倍体白杨杂种无性系的有效方法,能够有效鉴别21个三倍体白杨杂种无性系。本研究为品种鉴定及新品种权保护奠定了基础。     

Multilocus half-tetrad analysis and centromere mapping in citrus: evidence of SDR mechanism for 2n megagametophyte production and partial chiasma interference in mandarin cv 'Fortune'

The genetic structure of 2n gametes and, particularly, the parental heterozygosity restitution at each locus depends on the meiotic process by which they originated, with first-division restitution and second-division restitution (SDR) being the two major mechanisms. The origin of 2n gametes in citrus is still controversial, although sexual polyploidisation is widely used for triploid seedless cultivar development. In this study, we report the analysis of 2n gametes of mandarin cv 'Fortune' by genotyping 171 triploid hybrids with 35 simple sequence repeat markers. The microsatellite DNA allele counting-peak ratios method for allele-dosage evaluation proved highly efficient in segregating triploid progenies and allowed half-tetrad analysis (HTA) by inferring the 2n gamete allelic configuration. All 2n gametes arose from the female genitor. The observed maternal heterozygosity restitution varied between 10 and 82%, depending on the locus, thus SDR appears to be the mechanism underlying 2n gamete production in mandarin cv 'Fortune'. A new method to locate the centromere, based on the best fit between observed heterozygosity restitution within a linkage group and theoretical functions under either partial or no chiasmata interference hypotheses was successfully applied to linkage group II. The maximum value of heterozygosity restitution and the pattern of restitution along this linkage group would suggest there is partial chiasma interference. The implications of such a restitution mechanism for citrus breeding are discussed.     

Comparison of PCR-based molecular marker analyses of Musa breeding populations

Progress in the breeding of plantain and banana has been restricted by the complex genetic structure and behaviour of cultivated polyploid Musa. Genetic improvement has been hindered due to the large amount of space required for growth and maintenance of plant populations, in addition to the long growth cycle and the low levels of fertility and seed viability characteristic of cultivated genotypes. Molecular marker assisted breeding has the potential to dramatically enhance the pace and efficiency of genetic improvement in Musa. This study was conducted to compare different PCR-based marker systems (RAPD, VNTR and AFLP) for the analysis of breeding populations generated from two diverse Musa breeding schemes. All three assays detected a high level of polymorphism between parental genotypes and within progeny populations. As expected, AFLP assays had by far the highest multiplex ratio while VNTR analysis detected the highest levels of polymorphism. AFLP analysis of a full-sib tetraploid hybrid population confirmed previous reports based on VNTR analysis, of a high frequency of recombination during 2n (3x) gamete formation by a triploid plantain landrace. In addition, both VNTR and RAPD analyses of a full-sib triploid hybrid population suggested a high frequency of homoeologous recombination during n (2x) gamete formation by tetraploid hybrids. In general, there was a poor correlation between estimates of genetic similarity based on different types of marker. The implications of these findings for the molecular breeding of Musa crops are discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

Variation in pollen formation and its cytological mechanism in an allotriploid white poplar

Variation in pollen formation and its cytological mechanism in an allotriploid white poplar were investigated by the squashed technique and indirect immunofluorescence. Besides 0.5% stuck pollen grains, this allotriploid produced regularly spherical pollen grains. It was estimated that 90.3% of pollen grains were viable. Diameters of the viable spherical pollen grains ranged from 23.2 to 72.9 μm, with a bimodal frequency distribution. Numerous meiotic abnormalities were found, including highly irregular chromosome pairing, lagging chromosomes and chromosome bridges, micronuclei, and multiple spindles, which indicate highly genetic imbalance of this allotriploid. Some micronuclei triggered minispindle formation in metaphase II and participated in cytokinesis to form microcytes in sporads. Abnormal orientation of metaphase II spindles contributed to production of dyads and triads, which produced unreduced microspores. However, parallel orientation of spindles was not necessary for dyad formation, because an organelle band positioned in the equatorial region prevented the spindles from coalescing. Some microsporocytes exhibited a complete or partial absence of cytokinesis, which resulted in the formation of monads and the increased frequency of dyads and triads. The perspective of this triploid in the polyploid breeding program of white poplar is discussed.     

禾本科三倍体: 形成、鉴定与利用

禾本科三倍体的形成途径包括2n配子融合、倍性间杂交、多精受精和胚乳培养。其中, 2n配子融合和倍性间杂交分别为自然界和人工合成三倍体的主要途径。该文介绍了形态学观测、染色体分析、流式细胞术和分子标记等倍性鉴定方法在禾本科三倍体中的应用及其优缺点。目前, 三倍体在禾谷类作物中无直接应用价值, 但可作为通往多倍体、非整倍体和转移异源基因的遗传桥梁。多年生禾本科三倍体(特别是异源三倍体)在饲草或能源作物中已得到广泛应用, 在该类型禾本科作物中均可直接尝试三倍体育种。多倍体的三倍体育种和无融合生殖三倍体育种可作为未来禾本科三倍体的研究方向。三倍性胚乳培养可以一步合成三倍体, 多精受精可以实现遗传上3个不同基因组的一步融合, 在三倍体研究中应予以重视。鉴于2n配子融合、多精受精的稀有特性和倍性间杂交、胚乳培养频繁的染色体变异, 高通量三倍体鉴定技术的发展将是三倍体研究实现突破的关键。