异源六倍体水稻AACCDD和三倍体水稻ACD生殖特性的细胞胚胎学研究

利用异源多倍体杂种优势是多倍体水稻研究的第三阶段,但异源多倍体杂种常常不育。为明确其不育特点,本文以本实验室通过远缘杂交获得的栽培稻（AA）品种DTS137和高秆野生稻O．alta（CCDD）的杂种三倍体ACD和加倍形成的六倍体AACCDD为材料,分别对其花粉和胚囊发育过程进行石蜡切片观察,发现3x与6x之间以及6x雌性和雄性生殖方式和前途具有明显的不同：（1）异源三倍体ACD水稻杂种花粉败育彻底,败育发生在小孢子母细胞时期,绒毡层细胞提前解体：大孢子母细胞不能进行减数分裂,与周围的珠心组织一起发生解体,雌性完全败育。（2）异源六倍体水稻杂种（AACCDD）的雄性败育发生在小孢子母细胞减数分裂的细线期,此期小孢子母细胞发育停滞,随后解体;而雌性器官的发育基本正常。推测异源六倍体杂种的不育性与不同基因组间存在着部分核质不亲和性有关。据此,为了克服六倍体水稻AACCDD的不育性和验证该杂种雌性可育的结论,以栽培稻（AA）的PMeS二倍体品系HN2026．2x为父本与之杂交,通过胚挽救成功获得回交杂种BC1F1植株,经根尖染色体鉴定为2n=4x=48,系由AACD组成。虽然该异源三基四倍体是不育的,但为随后的染色体加倍创造AAAACCDD同源异源八倍体,进而获得结实的同源异源多倍体杂种打下了良好的基础。     

主要作物远缘杂交概况

本文综述了水稻、小麦、玉米、棉花、大豆等作物的远缘杂交的概况。这五种作物远缘杂交中,利用的野生近缘植物有水稻的尼瓦拉野生稻、普通野生稻和药用野生稻,小麦的黑麦、偃麦草、冰草、山羊草、簇毛麦、赖草、鹅观草、新麦草、旱麦草,玉米的类玉米和摩擦禾,棉花的墨西哥野生棉、异常棉、瑟伯氏棉、李奇蒙德氏棉、司笃克氏棉和比克氏棉,大豆的野生大豆和半野生大豆。通过远缘杂交获得了新物种如小偃麦、亚比棉等,还获得了大批异附加系、异代换系、易位系等种质材料,以及生产上推广的优良品种。     

鱼类遗传育种中生物学方法的应用及研究进展

鱼类遗传育种是指利用生物学方法对鱼类进行遗传选择或改造,从而获得新型改良鱼类的过程;它可以通过人为选择优势遗传性状或者通过整合或改变已有的遗传性状而达到遗传改良的目的.鱼类遗传育种是一个从稳定品系中筛选或研制变异品系,再从变异品系中培育稳定品系的循环过程.鱼类良种的培育成功将在很大程度上带动良种的饲养、加工、销售、休闲等后续产业的发展,这在渔业经济发展中具有重要意义.本文系统地总结了包括传统选择育种、分子标记辅助育种、全基因组选择育种和单性控制育种在内的选择育种技术,综述了杂交育种(近缘和远缘杂交)、细胞核移植、生殖干细胞和生殖细胞移植、人工雌核发育和雄核发育技术、多倍体育种在内的性状整合育种技术,以及以转基因育种为代表的性状改造育种技术,乃至这些育种技术在鱼类育种中的应用情况,同时结合本实验室在鱼类远缘杂交、雌核发育和雄核发育等染色体倍性育种方面的研究成果,对国内外鱼类育种的研究现状以及存在的问题进行了系统的概述.     

主要作物远缘杂交概况

本综述了水稻、小麦、玉米、棉花、大豆等作物的远缘杂交的概况。这五种作物远缘杂交中,利用的野生近缘植物有水稻的尼瓦拉野生稻、普通野生稻和药用野生稻,小麦的黑麦、偃麦草、冰草、山羊草、簇毛麦、赖草、鹅观草、新麦草、旱麦草,玉米的类玉米和摩擦禾,棉花的墨西哥野生棉、异常棉、瑟伯氏棉、李奇蒙德氏棉、司笆克氏棉和比克氏棉,大豆的野生大豆和半野生大豆。通过远缘杂交获得了新物种如小偃麦、亚比棉等,还获得了大批异附加系、异代换系、易位系等种质材料,以及生产上推广的优良品种。     

新型四倍体水稻创制及其杂种优势利用研究进展

新型四倍体水稻是本研究团队创制的一类高育性多倍体水稻新种质,携带有多种优异基因。该类材料与同源四倍体水稻杂交获得的杂种F₁产量优势明显,具有应用前景。本文首先概述了同源四倍体水稻的研究进展,并重点阐述了新型四倍体水稻创制过程、主要特点、杂种优势表现及其涉及的分子遗传机理,最后提出未来研究的思路及需要解决的问题,为加快新型四倍体水稻在多倍体水稻育种中的利用提供依据。     

两个具多倍体减数分裂稳定性的多倍体水稻品系的选育

多倍体化是植物进化的主要途径之一, 主要的粮棉油作物大都是多倍体, 它们在倍性增加的同时, 带来产量成倍增长. 在自然进化启示下, “利用远缘杂交和多倍体双重优势选育超级稻”是一个新途径. 但结实率低是一个制约多倍体水稻育种发展的瓶颈问题. 根据蔡得田等人提出的育种战略, 把研究重点放在类Ph基因材料选育上. 通过几年的籼粳杂交和回交选择与检测, 选育出2个具有多倍体减数分裂稳定性(polyploid meiosis stability, PMeS)的多倍体水稻品系PMeS-1, PMeS-2. 其选育过程包括亲本选择、杂交或复合杂交、连续回交、染色体加倍、多倍体鉴定、结实性选择、自交稳定成品系等7个步骤. 多倍体减数分裂稳定性的特性则通过减数分裂行为观察和高结实性杂交测定来确定. PMeS-1和PMeS-2都为粳型品系. 它们具有3个显著特点: 一是自身结实率较高, 超过65%; 二是与许多籼稻、粳稻多倍体品种杂交产生结实率高(>70%)、优势强的杂交后代; 三是表现出减数分裂稳定性, 即花粉母细胞减数分裂前期主要以二价体和四价体配对, 极少出现高价体、单价体和三价体, 中期很少出现落后染色体, 后期和末期未见微核. 而对照品系Dure-4X花粉母细胞减数分裂表现异常, 前期较高频率出现高价体、单价体和三价 体, 中期高频率出现落后染色体, 后期和末期有微核出现. 本研究建立了一种多倍体水稻选育的基本体系, PMeS品系选育方法与一般二倍体品系比较有三点不同, 即染色体加倍、多倍体检测 和高结实性检验. 利用PMeS品系基本上解决了多倍体水稻结实率低这个长期困扰多倍体水稻育种的难题, 为广大育种学家利用这种基因材料选育多倍体水稻新品种提供了良好的条件.     

植物远缘杂交中的染色体行为及其遗传与进化意义

远缘杂交与多倍体化在高等植物的进化中起着重要的作用。但矛盾和令人费解的现象是“自然界在合成多倍体方面取得了巨大的成功,而人类在这方面却收效甚微”。其原因一方面可能是自然多倍体是长期自然选择和进化的产物,人类难以在短期内重复和完成这一过程;另一方面可能对不同的染色体组结合后的遗传与互作机制还不太了解。故多倍体化后的遗传和表观遗传成了目前多学科研究的重点。在有些有性和体细胞杂种内亲本染色体在细胞内分开排列,但此染色体行为的遗传和生物学意义还不太清楚。在植物远缘杂交中出现的假配生殖、半配生殖、染色体消除和亲本染色体组分开等异常染色体行为,也反映出不同物种在配子和染色体水平上的不亲和。需对植物远缘杂交中的染色体行为和遗传进行不同层次与系统的研究,才可能深入了解杂交后新种的形成及进化机制。     

甘蓝型油菜与白菜型油菜杂交杂种第一代细胞学观察

远缘杂交是油菜育种中的重要途径。在油菜远缘杂交育种工作中,存在的主要问题之一是杂种植株的不育性。对于这个问题的研究,国内外已有一些报道,但从细胞学角度研究其实质还不多。为了更好的利用远缘杂交的办法创造新品种,有必要对该问题进行深入研究。本文对杂种植株花粉母细胞减数分裂的情况和花粉粒发育作了初步观察。     

从0到1:异源四倍体野生稻从头驯化创造全新作物

2021年2月3日,Cell杂志在线发表了李家洋院士团队关于四倍体野生稻从头驯化的重大突破性进展。该团队绘制了四倍体野生稻从头驯化路线图,与合作者经过多年攻关,突破了一系列限制多倍体野生稻驯化的理论难题和技术瓶颈,综合运用多维组学和基因编辑技术实现了野生四倍体水稻的从头驯化,创造了世界首例重新设计与快速驯化的四倍体水稻,将过去的“不可能”变成了现在的“一切皆有可能”。该研究突破了现有二倍体水稻作物的育种局限,实践了从0到1的突破性创新,为遗传背景复杂的野生植物重新驯化提供了范例和解决方案,将引领未来作物创造,翻开生物育种新篇章。     

作物远缘杂交育种的途径及其实质

作物远缘杂交的育种可操作性及效果多年来颇有争议,科学家对物种起源与进化的研究恰恰是指导作物远缘杂交育种的理论基础。物种形成理论研究表明生命的共同起源是远缘杂交的理论基础,生物多样性是远缘杂交的物质基础。生物种间的繁殖隔离机制是远缘杂交不亲和性障碍的根源所在,而物种形成方式又为克服远缘杂交的不亲和性提供了理论依据。其中异域性物种形成方式下的生殖隔离具有不彻底性,是克服远缘杂交受精前不亲和性的理论根据;同域性物种形成方式中多倍体化的方式对远缘杂交受精后不亲和性的克服具有较强的指导意义。本文在通过对以上方面的阐述,剖析了远缘杂交的障碍来源、克服途径及实质,为作物远缘杂交育种工作提供参考。     

我国杂交水稻基因工程育种策略探讨

杂交水稻在我国水稻及粮食生产中占有突出地位 ,基因工程可在杂交水稻育种中发挥重要作用。针对杂交水稻育种存在的主要问题 ,指出应将优质、抗虫和抗病作为当前基因工程育种的研究重点。同时提出基因工程育种与常规育种紧密结合 ,优质基因工程着重改良保持系及聚合转基因等策略 ,以培育出超高产优质抗病虫转基因聚合杂交稻新组合。     

Progress and prospects for interspecific hybridization in buckwheat and the genus Fagopyrum

Cultivated buckwheat, such as common (Fagopyrum esculentum Moench.) and tartary (Fagopyrum tataricum (L.) Gaertn.) buckwheat, is one of the most versatile crops for forage and food and has several benefits for human health. Interspecific hybridization between Fagopyrum species is of great importance to improvement of buckwheat. Hybridization would allow the transfer of agronomical beneficial characteristics from wild Fagopyrum species, including self-pollination and increased fertility, frost tolerance, and higher content of beneficial compounds. However, conventional breeding methods are only partially applicable because of the self-incompatibility and incompatibility barriers between different species. Present review summarizes the morphology of self-incompatibility, the genetic and cellular basis of incompatibility between different Fagopyrum species. In many interspecific crosses hybrid embryos are aborted after successful pollination due to post-zygotic incompatibility. The use of in vitro embryo rescue after interspecific hybridization has been successful in circumventing breeding barriers between Fagopyrum species. Methods applied successfully for the construction of interspecific hybrids are discussed in detail.     

籼稻二元不育系9730A及三交种的初步研究

通过对所选育的籼稻二元不育系9730A的花粉育性、自交结实率、雌性育性、对"920"的敏感性、柱头外露情况、花时、雄性不育的可恢复性及三交种的抽穗整齐度、有效穗、株高、单株产量、结实率、主茎穗长、主茎叶长、穗粒等变异度的分析,对杂交水稻三交法育种的应用价值进行初步探讨.结果认为:通过引进具有不同优良品种特性的第二保持系,可以在保持所选育杂种品种主要农艺性状整齐一致的前提下,较大幅度地拓宽品种的遗传基础,改进雄性不育系的异交性能,从而为提高杂种种子生产水平、及时利用优良常规稻育种成果材料和综合实现杂交稻品种选育过程中的多抗、广适、高产、优质育种目标提供新的途径.     

Rice interspecies hybrids show precocious or delayed developmental transitions in the endosperm without change to the rate of syncytial nuclear division

In angiosperms, interspecific crosses often display hybrid incompatibilities that are manifested as under‐proliferation or over‐proliferation of endosperm. Recent analyses using crosses between Arabidopsis thaliana and its related species with different ploidy levels have shown that interspecific hybridization causes delayed developmental transition and increased mitotic activity in the endosperm. In this study, we investigated endosperm development in interspecific crosses between diploid Oryza species. In a cross between female O. sativa and male O. punctata, we found that the hybrid endosperm was reduced in size and this cross was associated with precocious developmental transition. By contrast, the cross between O. sativa and O. longistaminata generated enlarged hybrid endosperm at the mid‐point of seed development and this cross was associated with delayed developmental transition. Subsequently, the hybrid endosperm displayed a shriveled appearance at the seed maturation stage. We found that the accumulation of storage products and the expression patterns of several marker genes were also altered in the hybrid endosperm. By contrast, the rate of syncytial mitotic nuclear divisions was not significantly affected. The gene OsMADS87 showed a maternal origin‐specific expression pattern in rice endosperm, in contrast to its Arabidopsis homologue PHERES1, which shows paternal origin‐specific expression. OsMADS87 expression was decreased or increased depending on the type of developmental transition change in the hybrid rice endosperm. Our results indicate that one of the interspecies hybridization barriers in Oryza endosperm is mediated by precocious or delayed developmental alterations and de‐regulation of OsMADS87, without change to the rate of syncytial mitotic nuclear division in the hybrid endosperm.     

Development of Anther- defective Cytoplasmic Male Sterile Line from Interspecific Hybridization Between Dongxiang Wild Rice and Cultivated Rice and Preliminary Observations on Its Derivatives

Exploring novel source of cytoplasmic male sterility (CMS) is essential to stablize the productivity of hybrid rice. Dongxiang wild rice ( Oryza rufipogon Griff. ) has been recorded as the northest distributed wild rice in China that is resistant to several biotic or abiotic stresses. A male sterile line M01A defective of anther was identified in the F3 population from an interspecific cross between Dongxiang wild rice and cultivated rice ( Oryza sativa L. ssp. indica ). Crosses and successive backcrosses were made between M01A and a variety of breeding materials and 19 progeny families were obtained. Among the families, some were defective of anthers, and some have twisted and degenerated anthers without microsporogenesis or with a few typical aborted pollen. These resuits implicate that the male sterility of M01A was genetically regulated by the interaction between the nucleus and the cytoplasm. Only one cross produced male-fertile hybrid in which the paternal parent contains a part of the genome of Dongxiang wild rice, which implies that Dongxiang wild rice itself could be the source of the fertility restorer to M01A.     

Natural variation for a hybrid incompatibility between two species of Mimulus

Understanding the process by which hybrid incompatibility alleles become established in natural populations remains a major challenge to evolutionary biology. Previously, we discovered a two-locus Dobzhansky-Muller incompatibility that causes severe hybrid male sterility between two inbred lines of the incompletely isolated wildflower species, Mimulus guttatus and M. nasutus. An interspecific cross between these two inbred lines revealed that the M. guttatus (IM62) allele at hybrid male sterility 1 (hms1) acts dominantly in combination with recessive M. nasutus (SF5) alleles at hybrid male sterility 2 (hms2) to cause nearly complete hybrid male sterility. In this report, we extend these genetic analyses to investigate intraspecific variation for the hms1-hms2 incompatibility in natural populations of M. nasutus and M. guttatus, performing a series of interspecific crosses between individuals collected from a variety of geographic locales. Our results suggest that hms2 incompatibility alleles are common and geographically widespread within M. nasutus, but absent or rare in M. guttatus. In contrast, the hms1 locus is polymorphic within M. guttatus and the incompatibility allele appears to be extremely geographically restricted. We found evidence for the presence of the hms1 incompatibility allele in only two M. guttatus populations that exist within a few kilometers of each other. The restricted distribution of the hms1 incompatibility allele might currently limit the potential for the hms1-hms2 incompatibility to act as a species barrier between sympatric populations of M. guttatus and M. nasutus. Extensive sampling within a single M. guttatus population revealed that the hms1 locus is polymorphic and that the incompatibility allele appears to segregate at intermediate frequency, a pattern that is consistent with either genetic drift or natural selection.     

中国野生稻遗传资源的保护及其在育种中的利用

我国有三种野生稻,即普通野生稻(Oryza rufipogon)、药用野生稻(O.officinalis)和瘤粒野生稻(O.meyeriana)。这三种野生稻均被列为国家二级保护植物(渐危种)。调查结果表明,野生稻由于其自然群落大量丧失而濒危,濒危程度为普通野生稻>药用野生稻>瘤粒野生稻。造成濒危的主要原因是人为的破坏活动。人类的经济活动导致了野生稻生境丧失、生境质量不断恶化、栖息地越来越少;人类的活动也导致了外来种的入侵。目前,对野生稻的保护措施主要有就地保护(原地保护或原位保护)和迁地保护(易地保护或异位保护)。易地保护包括以种子保存的种质厍、以种茎保存的种质圃和以器官培养物作为材料的超低温保存。野生稻具有许多优良特性,如特强的耐寒性、高的抗病虫性、优质蛋白质含量高、功能叶片耐衰老的特异性、特强的再生性、良好的繁茂性及生长优势等等,这些优良特性已被广泛用于水稻常规育种和杂交育种中,并取得了巨大的社会效益和经济效益。有关野生稻生物技术方面的研究,如花药培养、原生质培养、体细胞杂交和基因工程等方面已取得了较大的进展。野生稻将在水稻育种中发挥越来越重要的作用。     

中国水稻遗传育种历程与展望

我国的水稻育种经历了矮化育种、杂种优势利用和绿色超级稻培育3次飞跃,其间伴随矮化育种(第一次绿色革命)、三系杂交稻培育、二系杂交稻培育、亚种间杂种优势利用、理想株型育种和绿色超级稻培育等6个重要历程。育种目标从唯产量是举到高抗、优质和高产并重,育种理念从高产优质逐步提升为“少投入,多产出,保护环境”。水稻功能基因组研究为第二次绿色革命准备了大量的有重要利用价值的基因,水稻育种正迈向设计育种的新时代。基因组选择技术和转基因技术将为培育“少打农药,少施化肥,节水抗旱,优质高产” 绿色超级稻保驾护航。本文对我国水稻遗传育种的发展历程进行了概括,指出了各种育种方法和育种技术的优缺点,系统介绍了水稻细胞质雄性不育和光温敏雄性核不育以及籼粳杂种不育的分子机制的研究进展,综述了水稻株型、穗型、粒形和养分高效利用相关的重要功能基因,阐明了产量与开花期联动的关系,凸显了我国水稻基础研究在国际上的重要地位。特别指出,近年来,我国水稻生产方式发生了或正在发生巨大变革,育种理念也要与时俱进。未来,杂交育种技术要与现代育种技术紧密结合,选育水稻品种不仅要满足市场需求,而且更要具备绿色健康的特点,同时还要适应新耕作制度和新耕作方法。     

Understanding the genetic and molecular constitutions of heterosis for developing hybrid rice

The wide adoption of hybrid rice has greatly increased rice yield in the last several decades. The utilization of heterosis facilitated by male sterility has been a common strategy for hybrid rice development. Here, we summarize our efforts in the genetic and molecular understanding of heterosis and male sterility together with the related progress from other research groups. Analyses of F₁ diallel crosses show that strong heterosis widely exists in hybrids of diverse germplasms, and inter-subspecific hybrids often display higher heterosis. Using the elite hybrid Shanyou 63 as a model, an immortalized F₂ population design is conducted for systematic characterization of the biological mechanism of heterosis, with identification of loci controlling heterosis of yield and yield component traits. Dominance, overdominance, and epistasis all play important roles in the genetic basis of heterosis; quantitative assessment of these components well addressed the three classical genetic hypotheses for heterosis. Environment-sensitive genic male sterility (EGMS) enables the development of two-line hybrids, and long noncoding RNAs often function as regulators of EGMS. Inter-subspecific hybrids show greatly reduced fertility; the identification and molecular characterization of hybrid sterility genes offer strategies for overcoming inter-subspecific hybrid sterility. These developments have significant implications for future hybrid rice improvement using genomic breeding.     

Hybrid breeding of rice via genomic selection

Hybrid breeding is the main strategy for improving productivity in many crops, especially in rice and maize. Genomic hybrid breeding is a technology that uses whole‐genome markers to predict future hybrids. Predicted superior hybrids are then field evaluated and released as new hybrid cultivars after their superior performances are confirmed. This will increase the opportunity of selecting true superior hybrids with minimum costs. Here, we used genomic best linear unbiased prediction to perform hybrid performance prediction using an existing rice population of 1495 hybrids. Replicated 10‐fold cross‐validations showed that the prediction abilities on ten agronomic traits ranged from 0.35 to 0.92. Using the 1495 rice hybrids as a training sample, we predicted six agronomic traits of 100 hybrids derived from half diallel crosses involving 21 parents that are different from the parents of the hybrids in the training sample. The prediction abilities were relatively high, varying from 0.54 (yield) to 0.92 (grain length). We concluded that the current population of 1495 hybrids can be used to predict hybrids from seemingly unrelated parents. Eventually, we used this training population to predict all potential hybrids of cytoplasm male sterile lines from 3000 rice varieties from the 3K Rice Genome Project. Using a breeding index combining 10 traits, we identified the top and bottom 200 predicted hybrids. SNP genotypes of the training population and parameters estimated from this training population are available for general uses and further validation in genomic hybrid prediction of all potential hybrids generated from all varieties of rice.