东乡野生稻与栽培稻正反交种间杂种F1的雄配子发生与发育

该文以东乡野生稻与栽培稻(美国光壳稻P1003)的正反交种间杂种F₁(正交为光壳稻P1003×东乡野生稻; 反交为东乡野生稻×光壳稻P1003)为试材, 研究其各个减数分裂时期的染色体行为特征、染色体交换重组及雄配子发育特点。结果表明: 正反交杂种F₁花粉母细胞细胞核减数分裂的同步性较高, 细胞质为连续型胞质分裂类型。在细胞核分裂的过程中, 核仁在前期I到中期I逐渐消失。染色体在前期I到中期I逐渐收缩, 变得更加清晰可见。在终变期和中期I, 90.54%以上的花粉母细胞能形成12个二价体(含少数棒状二价体和8字型二价体), 部分花粉母细胞(正交9.46%, 反交7.65%)出现少量的单价体、三价体和四价体。后期I观察到1.27%–1.35%的花粉母细胞含有1至数条落后染色体。最终有92.6%–94.8%的小孢子能发育成形态正常、染色能力强的成熟花粉粒。另外, 在正交杂种F₁的粗线期中存在2个核仁, 而反交杂种F₁及其双亲的粗线期只有1个核仁。这些研究结果可为作物品种改良、种质资源创新以及种间亲缘关系研究提供细胞学证据。     

东乡野生稻耐冷渐渗系的细胞学观察及SSR分析

以前期鉴定筛选的2个东乡野生稻强耐冷渐渗系(IL5243和IL5335)为试材,研究其减数分裂时期的染色体行为特征及外源基因的渗入分子证据。结果表明:(1)IL5243和IL5335中正常减数分裂的花粉母细胞分别达89.93%和90.22%,最终形成正常的成熟花粉粒,花粉离体萌发率分别为(83.03±2.82)%和(81.96±1.73)%,与受体亲本无显著性差异。(2)在减数分裂I中,2个耐冷渐渗系均观察到低频率异常染色体行为,如单价体、"8"字型二价体、多价体,以及后期I有少数花粉母细胞(3.95%～5.15%)存在落后染色体等,表明其染色体组之间发生了交换和重组;在粗线期,2个强耐冷渐渗系均观察到较高频率(IL5243和IL5335分别为27.0%和38.9%)的双核仁,而其双亲都是单核仁。(3)SSR标记和Structure分析进一步证实了栽培稻和野生稻染色体组间发生了交换重组,东乡野生稻部分DNA片段已渗入到强耐冷渐渗系中,这为水稻耐冷基因的挖掘与利用奠定了重要基础。     

番茄属种间杂种离体培养F1植株的细胞学分析

本文报道了栽培番茄（Lycopersico esculentum)“北京早红”等5个品种分别与野生型秘鲁番茄（L.peruvianum)PI128657中8号株系杂交,离体胚培养,得F1杂种植株,对花粉母细胞在减数分裂中染色体行为和终变期二价体交叉点的频率,以及亲和性程度等进行了.结果表明,6个亲本植株花粉母细胞减数分裂染色体的行为是正常的,中期I为12个二价体。其中环状二价体占多数,棒状二价体数较少,中期I没有单价体,后期I和II均正常,四分体阶段无微出现,但各亲本在终变期和中期I的环状二价体和棒状二价体的数有一定的差异,这可能与不同亲本基因型的亲和性程度和在遗传学上的不协调有关。5个组合的大多数F1杂种花粉母细胞减数分裂中染色体行为基本正常,12个二价体占多数,但染色体配对不稳定,有较多的单价体,染色单体桥。四分体阶段有微核,此外,在5个组合的F1杂种植株中,均出现双二倍体花粉母细胞,这些双二倍体花粉母细胞的染色体,在减数分裂中,也均,出现落后染色体和染色单体桥,以及较多的多价体,四分体阶段有微和不同类型的四分孢子群。     

辣椒属种间杂种F1植析的细胞遗传学研究

本文分析了早椒和Ｃ;ｈｉｎｅｎｓｅ 二亲本及Ｆ１杂种花粉母细胞减数分裂终变期和中期Ⅰ期染色体配对的构型。结果表明;二亲本工粉母细胞减娄分裂染色体配对均正常,为１２个二价体。Ｆ１杂种花粉母细胞减数分裂染色体配对很不规则。其平均频率为单价体０．０３６,二价体９．１８,三价体０．０３６,四价体０．８０,六价体０．３８。     

黄瓜花粉母细胞减数分裂行为的研究

研究了华北型、华南型和西南型黄瓜花粉母细胞的减数分裂行为,发现黄瓜细胞核减数分裂的同步性较高,细胞质是同时型分裂。在细胞核分裂的过程中,核仁在前期Ⅰ到中期Ⅰ逐渐消失,在前期Ⅰ再次出现,随后消失;染色体在前期Ⅰ到中期Ⅰ逐渐收缩,变得清晰,至末期Ⅰ解螺旋,变得模糊,在前期I再次清晰。不同生态型黄瓜终变期的染色体构型均以环状二价体为主。在前期Ⅰ和前期Ⅰ,西双版纳黄瓜的核仁都相应地比另外两种生态型黄瓜品种的多,在后期Ⅰ还偶尔出现染色体桥,显示了西双版纳黄瓜变种的特殊性。研究还发现寒冷的气候条件下栽培黄瓜都能够形成高频率的多分体,推测其形成很可能与低温逆境有关。     

大鹅观草与蛇河披碱草属间杂种的细胞遗传学研究

为探讨大鹅观草(Roegneria grandis,2n=4x=28)的染色体组组成,为其正确的分类处理提供细胞学依据。该研究通过人工远缘杂交,成功获得3株大鹅观草与蛇河披碱草(Elymus wawawaiensis,2n=4x=28)属间杂种F1植株。杂种植株形态介于两亲本之间,不育。亲本及杂种经I2-IK溶液染色后进行花粉育性检测,结果显示Roegneria grandis和Elymus wawawaiensis的花粉可育,育性高达94.6%和90.5%;杂种F1不育。花粉母细胞减数分裂中期I染色体配对结果显示,亲本花粉母细胞配对正常,均形成14个二价体,以环状二价体为主,Roegneria grandis有频率很低(0.04/细胞)的单价体出现;杂种F1平均每个花粉母细胞形成6.46个二价体,变化范围为5~8;在观察的83个花粉母细胞中,有35.2%的花粉母细胞形成了7个二价体,形成6个二价体的细胞占42.59%,较少细胞形成8个二价体;平均每个细胞形成14.66个单价体,变化范围为10~18;平均每细胞观察到0.14个三价体;杂种花粉母细胞染色体构型为14.66 I+6.46 II+0.14 III;平均每细胞交叉数为9.83,C值为0.35。结果表明:(1)R.grandis与Elymus wawawaiensis有一组染色体组同源的St染色体组,另外一组染色体组不是St或者H染色体组,Roegneria grandis的染色体组组成不是St Stg;(2)较低频率的三价体(平均0.14个/细胞),可能是由于R.grandis的St和Y染色体组间具有一定的同源性,也可能是染色体易位等原因导致,对于Y染色体组的起源还需深入地研究;(3)在不同地理来源的披碱草属和鹅观草属物种中St染色体组同源性不同,R.grandis与来自于北美的Elymus lanceolatus与E.wawawaiensis的St染色体组较与分布于亚洲的E.sibiricus和E.caninus的St染色体组同源性反而更高,其原因还需要进一步地研究。     

中国东乡野生稻的粗线期分析及其与普通栽培稻的亲和性

中国东乡野生稻具多年宿根性,生长习性分直立型和匍匐型二类,群体内已有基因分化。形态性状分析证明其为普通野生稻的生态型变种。根尖细胞学鉴定,其染色体数为2n=24。花粉母细胞减数分裂行为正常。粗线期分析证实:该种具3个中部着丝点染色体,8个次中部着丝点染色体及一个亚端部着丝点染色体。染色体ⅤⅢ与核仁相连。染色体绝对长度为64.4—18.7μ。与栽培稻杂交育性正常,染色体配对良好,染色体组型可能为AA。     

Iris japonica×Iris confusa种间杂种的细胞遗传学研究

对Irisjaponica、I confusa和它们的人工种间杂种花粉母细胞减数分裂中期Ⅰ染色体配对行为、形态学和繁育特征进行了分析 ,结果表明 :(1)I japonica和I confusa的形态差异较小 ,杂种F1形态介于亲本之间 ;(2 )人工杂交比较容易 ,杂种种子能正常发育 ;杂种F1体细胞染色体数为 2n =30 ,减数分裂中期I染色体配对频率很高 ,为 13 9个二价体 ,构型为0 95Ⅰ + 5 95Ⅱ (棒形 ) + 7 95Ⅱ (环形 ) + 0 0 5Ⅲ + 0 2 7Ⅳ ,表明它们有相似的染色体组 ,亲缘关系很近 ;(3)杂种中有少量的三价体和四价体存在 ,可能是亲本染色体间发生了结构重排或是部分同源染色体配对 ;(4)杂种中大多数细胞存在 2个单价体 ,有的高达 8个单价体 ;花粉育性为 5 0 5 1% ,不能正常结实 ,表明I japonica和I confusa间存在生殖隔离 ,是独立的生物学物种 ;(5 )杂种F1减数分裂前期可观察到细胞融合 ,这可能是造成I japonica和I confusa不同居群出现多倍体和非整倍体的重要原因。     

甜瓜花粉母细胞减数分裂及雄配子体的细胞学研究

以薄皮和厚皮类型甜瓜为试材,采用改良的染色体制片方法,系统观察了甜瓜花粉母细胞的减数分裂及雄配子体发育的过程,结果表明:(1)甜瓜细胞核减数分裂的同步性较高,细胞质是同时型分裂,在细胞核分裂的过程中,核仁在前期Ⅰ到中期Ⅰ逐渐消失,在前期Ⅱ再次出现,随后消失,染色体在前期Ⅰ到中期Ⅰ逐渐收缩,变得清晰,至末期Ⅰ变得模糊,在前期Ⅱ再次清晰;(2)2种类型甜瓜终变期的染色体构型都以环状二价体为主;(3)在后期Ⅱ,观察到染色体的垂直和平行2种分离方式;(4)在前期Ⅰ和前期Ⅱ,伽师瓜"形成了多个较小的核仁,呈现一定的特殊性;(5)雄配子体发育经历了单核期和双核期,最后形成了成熟的花粉粒.研究表明,薄皮和厚皮类型甜瓜减数分裂的染色体行为基本一致,没有明显差异;伽师瓜"的核仁数量表现特殊可能与其长期的生态适应性有关.     

广东万年青的细胞遗传学研究

采用常规压片法和去壁低渗法,以生长于广西凭祥和广东深圳的广东万年青（Aglaonema modestumSchott ex Engl.）为材料,对二者的体细胞染色体、花粉母细胞减数分裂染色体配对行为和花粉发育过程进行了观察。结果表明：（1）野生植株为二倍体2n=40,栽培植株为三倍体2n=60;（2）野生种的小孢子母细胞减数分裂前期I终变期均为二价联会,栽培种偶见三价联会;（3）中期I,野生种为20个二价体排列在赤道板上,未见单价体,栽培种的20个二价体排列于赤道板上,20个单价体随机分布于两极,证实其为三倍体;（4）后期I,野生种二价体分离,出现单染色单体桥和断片,几率为10%,栽培种几率为3%,还存在落后染色体,部分不能进入两极的落后染色体和染色体断片在末期I形成微核;（5）四分体时期,野生种未观察到异常孢子,栽培种出现大量败育的四分体或多分体;（6）小孢子进入正常的发育分化,通过两次有丝分裂形成三细胞型花粉。野生种成熟花粉败育率为2.18%,栽培种为88.29%;（7）野生种正常结实,栽培种果实中未发现种子,为高度不育。     

Characterization of Interspecific Hybrids Between Oryza sativa L. and Three Wild Rice Species of China by Genomic In Situ Hybridization

In the genus Oryza, interspecific hybrids are useful bridges for transferring the desired genes from wild species to cultivated rice （Oryza sativa L.）. In the present study, hybrids between O. sativa （AA genome） and three Chinese wild rices, namely O. rufipogon （AA genome）, O. officinalis （CC genome）, and O. meyeriana （GG genome）, were produced. Agricultural traits of the F1 hybrids surveyed were intermediate between their parents and appreciably resembled wild rice parents. Except for the O. sativa × O. rufipogon hybrid, the other F1 hybrids were completely sterile. Genomic in situ hybridization （GISH） was used for hybrid verification. Wild rice genomic DNAs were used as probes and cultivated rice DNA was used as a block. With the exception of O. rufipogon chromosomes, this method distinguished the other two wild rice and cultivated rice chromosomes at the stage of mitotic metaphase with different blocking ratios. The results suggest that a more distant phylogenetic relationship exists between O. meyeriana and O. sativa and that O. rufipogon and O. sativa share a high degree of sequence homology. The average mitotic chromosome length of O. officinalis and O. meyeriana was 1.25- and 1.51-fold that of O. sativa, respectively. 4＇,6＇-Diamidino- 2-phenylindole staining showed that the chromosomes of O. officinalis and O. meyeriana harbored more heterochromatin, suggesting that the C and G genomes were amplified with repetitive sequences compared with the A genome. Although chromocenters formed by chromatin compaction were detected with wild rice-specific signals corresponding to the C and G genomes in discrete domains of the F1 hybrid interphase nuclei, the size and number of O. meyeriana chromocenters were bigger and greater than those of O. officinalis. The present results provide an important understanding of the genomic relationships and a tool for the transfer of useful genes from three native wild rice species in China to cultivars.     

Simple sequence repeat analyses of interspecific hybrids and MAALs of <Emphasis Type="Italic">Oryza </Emphasis><Emphasis Type="Italic">officinalis</Emphasis> and <Emphasis Type="Italic">Oryza sativa</Emphasis>

Wild rice is a valuable resource for the genetic improvement of cultivated rice (Oryza sativa L., AA genome). Molecular markers are important tools for monitoring gene introgression from wild rice into cultivated rice. In this study, Simple sequence repeat (SSR) markers were used to analyze interspecific hybrids of O. sativa-O. officinalis (CC genome), the backcrossing progenies and the parent plants. Results showed that most of the SSR primers (335 out of 396, 84.6%) developed in cultivated rice successfully amplified products from DNA samples of wild rice O. officinalis. The polymorphism ratio of SSR bands between O. sativa and O. officinalis was as high as 93.9%, indicating differences between the two species with respect to SSRs. When the SSR markers were applied in the interspecific hybrids, only a portion of SSR primers amplified O. officinalis-specific bands in the F(1) hybrid (52.5%), BC(1) (52.5%), and MAALs (37.0%); a number of the bands disappeared. Of the 124 SSR loci that detected officinalis-specific bands in MAAL plants, 96 (77.4%) showed synteny between the A and C-genomes, and 20 (16.1%) showed duplication in the C-genome. Sequencing analysis revealed that indels, substitution and duplication contribute to the diversity of SSR loci between the genomes of O. sativa and O. officinalis.     

Fitness estimation through performance comparison of F1 hybrids with their parental species Oryza rufipogon and O. sativa

BACKGROUND AND AIMS: Introgression of crop genes into populations of wild relatives has important implications for germplasm conservation as well as for the persistence of novel transgenes in wild populations. Studies of hybrid fitness can be used to evaluate the potential for introgression to occur following episodes of interspecific hybridization. METHODS: This study estimated relative fitness of interspecific hybrids through performance comparison of F(1) hybrids with their parental species, a cultivated rice (Oryza sativa) Minghui-63 and perennial common wild rice (O. rufipogon) under the cultivation conditions. KEY RESULTS: Compared with their parents, the hybrids had the lowest values of seedling survival ability, pollen viability and seed production; intermediate values of seed germination, spikelet production and flag leaf areas; and the highest values of plant height, number of tillers and panicles. The hybrids performed poorly at the stage of sexual reproduction, although they had a slightly higher hybrid vigour at the vegetative growth stage and better tillering ability than their wild parent. There were no significant differences in composite fitness across the whole life-history between the hybrids and their wild parental species. CONCLUSIONS: Rice genes, including transgenes, might persist in wild rice populations through vegetative and sexual reproduction. Further studies are needed to examine whether the extent of gene flow from rice is sufficiently significant to influence genetic diversity in wild populations of O. rufipogon, a species that has become endangered in some regions of south-east Asia.     

Distribution of fertility-restorer genes for wild-abortive and Honglian CMS lines of rice in the AA genome species of genus Oryza

BACKGROUND AND AIMS: Rice (Oryza sativa) is one of the most important cereal plants in the world. Wild-abortive (WA) and Honglian (HL) cytoplasmic male sterility (CMS) have been used extensively in the production of hybrid seeds. Although a variable number of fertility-restorer genes (Rf) for WA and HL-CMS have been identified in various cultivars, information on Rf in Oryza species with the AA-genome is sparse. Therefore the distribution and heredity of Rf for WA and HL-CMS in wild rice species of Oryza with the AA-genome were investigated. METHODS: Fertility-restorer genes for WA and HL-CMS in wild rice species with the AA-genome were investigated by following the fertility of microspores identified by I2-KI staining and by following the seed-setting rate of spikelets. A genetic model of Rf in some selected restorer accessions was analysed based on the fertility segregation of BC1F1 populations. KEY RESULTS: Fertility analysis showed that 21 out of 35 HL-type F1s, and 13 out of 31 WA-type F1s were scored as fertile. The frequency of Rf in wild rice was 60% for HL-CMS and 41.9% for WA-CMS, respectively. The fertility-restorer accessions, especially those with complete restoring ability, aggregated mainly in two species of O. rufipogon and O. nivara. The wild rice accessions with Rf for HL-CMS were distributed in Asia, Oceania, Latin American and Africa, but were centered mainly in Asia, whilst the wild restorer accessions for WA-CMS were limited only to Asia and Africa. Apart from one restorer accession that possessed two pairs of Rf for WA-CMS, all of the other nine tested wild restorer accessions each contained only a single Rf for WA-CMS or HL-CMS. Allele analysis indicated that there existed at least three Rf loci for the WA and HL-CMS systems. CONCLUSIONS: These data support the hypothesis that fertility-restorer genes exist widely in Oryza species with the AA-genome, and that Rf in Oryza sativa originated from the Oryza rufipogon/Oryza nivara complex, the ancestor of cultivated rice in Asia. The origin and evolution of Rf is tightly linked to that of CMS in wild rice, and fertility of a given CMS type is controlled by several Rf alleles in various wild restorer accessions.     

杂草稻红色果皮基因的遗传分析

以江苏扬中红色果皮杂草稻W16、粳型广亲和品种02428(S5n)及以它们为亲本的衍生世代F1、F2和F3为材料,研究了杂草稻红色果皮性状的遗传特征;根据已克隆的普通野生稻(O.rufipogen)红色果皮Rc与白色果皮rc等位基因第6外显子的差异,设计了InDel标记RID14,利用RID14标记对F2群体、江淮流域所收集到的24份红色果皮杂草稻、3份普通野生稻以及423份品种资源进行了标记基因型分析;选取5份江淮流域杂草稻的RID14标记PCR产物进行测序分析.结果显示:以白色果皮02428为母本的杂交种F1的颜色为白色,而以红色果皮杂草稻W16为母本,杂交种F1的果皮颜色为红色;正反交杂种F1所结F2种子都是红色,F2植株所结F3种子果皮颜色发生分离,符合3∶1分离比例;在F2群体中,RID14标记与果皮颜色共分离,在450份材料中,红色果皮均为非缺失带型,而白色果皮和紫色果皮为缺失带型;5份江淮流域杂草稻的RID14标记PCR产物序列与已登录的普通野生稻序列完全一致.研究表明,扬中杂草稻红色果皮为单基因控制的显性性状,并由母体基因型决定,是典型的延迟遗传,由Rc基因控制;在白色果皮材料资源中都存在14 bp缺失的等位基因rc,RID14标记可以作为准确鉴定红色果皮杂草稻的分子标记.     

Genetics of Hybrid Sterility and Hybrid Breakdown in an Intersubspecific Rice (Oryza Sativa L.) Population

F(1) hybrid sterility and ``hybrid breakdown' of F(2) and later generations in rice (Oryza sativa L.) are common and genetically complicated. We used a restriction fragment length polymorphism linkage map and F(4) progeny testing to investigate hybrid sterility and hybrid breakdown in a cross between ``widely compatible' O. sativa ssp. japonica cultivar Lemont from the Southern U.S. and ssp. indica cultivar Teqing from China. Our results implicate different genetic mechanisms in hybrid sterility and hybrid breakdown, respectively. Hybrid sterility appeared to be due to recombination within a number of putative differentiated ``supergenes' in the rice genome, which may reflect cryptic structural rearrangements. The cytoplasmic genome had a large effect on fertility of both male and female gametes in the F(1) hybrids. There appeared to be a pair of complementary genes that behaved like ``wide compatibility' genes. This pair of genes and the ``gamete eliminator' (S(1)) or ``egg killer' (S-5) may influence the phenotypic effects of presumed supergenes in hybrids. Hybrid breakdown appeared to be largely due to incompatibilities between indica and japonica alleles at many unlinked epistatic loci in the genome. These proposed mechanisms may partly account for the complicated nature of postreproductive barriers in rice.     

Crossability Barriers in the Interspecific Hybridization between Oryza sativa and O. meyeriana

Oryza meyerlana Baill (GG genome) Is a precious germplaem in the tertiary gene pool of cultivated rice (AA genome), and possesses important traits such as resistance and tolerance to biotic and abiotic stress. However, interspecific crossability barrier, a critical bottleneck restricting genes transfer from O. meyeriana to cultivars has led to no hybrids through conventional reproduction. Therefore, the reasons undedying incrossability were investigated in the present report. The results showed that: (ⅰ) at 3-7 d after pollination (DAP), many hybdd embryos degenerated at the earlier globular-shaped stage, and could not develop into the later pear-shaped stage. Meanwhile, free endosperm nuclei started to degenerate at 1 DAP, and cellular endosperm could not form st 3 DAP, leading to nutrition starvation for young embryo development; (ⅱ) st 11-13 DAP, almost all hybrid ovaries aborted. Even though 72.22% of hybrid young embryos were produced in the interspecific hybridization between O. sativa and O. meyeriana, young embryos were not able to further develop into hybrid plantlets via culturing in vitro. The main reason for the incrossability was hybrid embryo inviability, presenting as embryo development stagnation and degeneration since 3 DAP. Some possible approaches to overcome the crossability banders in the interspecific hybridization between O. sativa and O. meyeriana are discussed.     

QTL mapping and introgression of yield-related traits from <Emphasis Type="Italic">Oryza glumaepatula</Emphasis> to cultivated rice (<Emphasis Type="Italic">Oryza sativa</Emphasis>) using microsatellite markers

Rice ( Oryza sativa) cultivar development currently faces the task of overcoming yield plateaus, which is difficult due to the narrow genetic base of breeding programs. Oryza glumaepatula is a diploid wild relative of cultivated rice, native to Central and South America, and is therefore a potential source of alleles of agronomic importance to rice breeding programs. We studied 11 agronomic traits in BC(2)F(2) families of the interspecific cross Oryza sativa x O. glumaepatula. Transgressive lines which are almost isogenic to the elite recurrent O. sativa parent were identified for most of these traits. Quantitative trait locus (QTL) analysis was performed by single-point and interval mapping using a molecular map based on 157 microsatellite and STS markers. Marker regions accounting for 14.5 to 72.9% of a phenotypic variation trait were identified in 9 of the 12 rice chromosomes. Positive QTL effects from O. glumaepatula were observed in chromosomal regions associated with tillering and panicle-number traits.     

栽培黄瓜与野黄瓜正反杂交的几种同工酶分析

运用天冬氨酸转氨酶(AAT)、苹果酸脱氢酶(MDH)以及酯酶(EST)3种同工酶对栽培黄瓜"长春密刺"Cucumis sativus cv. Changchunmici (2n=14)与野黄瓜C. hystrix (2n=24)的正反交种间杂种F1 (正交: 野黄瓜×栽培黄瓜"长春密刺", 反交: 栽培黄瓜"长春密刺"×野黄瓜)及其双亲进行鉴定和比较研究。结果表明: 正反交种间杂种F1主要表现为双亲酶带的互补, 同时还形成4个杂合带(Aat-1-94、Aat-2-104、Mdh-3-102和Est-5-102)。上述3种酶均能准确地鉴定种间杂种的真实性。研究还发现正反交杂种F1的AAT和MDH的酶谱分别在酶带数目和强弱上表现出一定的差异, 进一步证实了野黄瓜与栽培黄瓜杂交存在正反交差异。