同源四倍体高粱与约翰逊草杂交的细胞遗传学研究

对约翰逊草、人工诱变四倍体高粱品系四沈甜及二者杂交种中期Ⅰ染色体构型、后期Ⅰ染色体行为进行了观察,并对花粉育性与结实率关系进行了研究。结果表明：约翰逊草、四沈甜及杂交种的染色体构型分别是：0．49Ⅰ＋15．83Ⅱ＋0．15Ⅲ＋1．60Ⅳ、0．72Ⅰ＋15．23Ⅱ＋0．075Ⅲ十2．15Ⅳ、0．68Ⅰ＋17．00Ⅱ＋0．18Ⅲ＋0．95Ⅳ。双亲及杂交种都是不规则的四倍体遗传群体。约翰逊草与同源四倍体高粱的染色体组间存在一定程度的同源性,杂交容易成功。     

四倍体鲫鲤、三倍体湘云鲫染色体减数分裂观察

用精巢细胞直接制片法观察了异源四倍体鲫鲤、三倍体湘云鲫和二倍体红鲫、湘江野鲤精母细胞染色体第一次减数分裂中期配对情况 ;作为对照 ,观察了上述四种鱼肾细胞的有丝分裂中期染色体。在精母细胞第一次减数分裂中 ,异源四倍体鲫鲤同源染色体两两配对 ,形成 10 0个二价体 ,没有观察到单价体、三价体和四价体 ;三倍体湘云鲫精母细胞形成 5 0个二价体和 5 0个单价体 ;红鲫和湘江野鲤精母细胞分别形成 5 0个二价体。肾细胞检测表明异源四倍体的染色体数目为 4n =2 0 0 ;湘云鲫为 3n =15 0 ;红鲫和湘江野鲤分别为 2n =10 0。减数分裂时染色体分布情况与肾细胞染色体检测结果相吻合。具有四套染色体的异源四倍体鲫鲤在减数分裂中只形成 10 0个二价体 ,而不形成 2 5个四价体或其它形式 ,为产生稳定一致的二倍体配子提供了重要的遗传保障 ,也为人工培育的异源四倍体鲫鲤群体能够世世代代自身繁衍下去提供了重要的遗传学证据。三倍体湘云鲫在减数分裂过程中出现二价体、单价体共存 ,同源染色体在配对和分离中出现紊乱 ,导致非整倍体生殖细胞的产生 ,为湘云鲫的不育性提供了染色体水平上的证据     

云南山茶花四倍体的首次发现及其科学意义

本文对分布于云南和四川金沙江河谷的云南山茶花C. reticulata及其两个近缘种(怒江山茶C.saluenensis和西南山茶C．pitardii)进行了细胞学研究。34个居群的云南山茶花中,21个居群是四倍体类型(2n=60),11个居群是六倍体类型(2n=90),另2个居群为二倍体(2n=30),云南山茶花的四倍体类型为首次发现,并且进行了核形态研究。四倍体和六倍体的花粉母细胞减数分裂染色体构型在大多数居群都为二价体(四倍体中30个二价体,六倍体中45个二价体),少数居群或是个体除二价体为主外还出现单价体和四价体,六倍体类型没有出现六价体构型。根据减数分裂的构型,我们认为,四倍体和六倍体分别为异源四倍体和异源六倍体,少数四价体的存在表明染色体有部分同源性。所有四倍体和六倍体的体细胞间期核特征和前期染色体形态特征基本相似。中海拔(1800m)以上的四倍体的云南山茶花外部形态特征与六倍体类型比较相似,而低海拔(1100～1800 m)的四倍体类型的外部形态特征则有些不同,但核形态结构是比较相似的。四倍体与六倍体类型地理分布是连续的,并与近缘的二倍体种怒江山茶、西南山茶重叠分布。     

拟南芥同源四倍体减数分裂过程的分子细胞学分析

以拟南芥(Columbia生态型)二倍体(AA,2n=10)为材料,经0.2%秋水仙素处理和细胞学鉴定,成功获得拟南芥同源四倍体(AAAA,2n=20)。以二倍体为对照,通过对拟南芥同源四倍体减数分裂过程染色体行为的观察,以及减数分裂调控同源染色体联会与重组相关基因的定量PCR分析,研究结果表明,与二倍体相比,拟南芥同源四倍体叶片表皮细胞间气孔孔径显著增大,荚果变长,但气孔密度和结实率显著降低;在减数分裂过程中出现部分单价体和三价体,以及二价体和四价体等染色体配对构型;减数分裂期重组相关基因ZYP1表达水平降低,ASY1、DMC1、MRE11和SPO11-1表达水平均升高。因此,我们推断,伴随着多倍体化,与二倍体相比,多倍体植物减数分裂期染色体行为和相关基因表达都有一定改变,影响多倍体植物生殖发育以适应环境。     

同源四倍体矮牵牛花粉母细胞减数分裂观察

以同源四倍体矮牵牛06P-12为材料,采用常规压片法对花粉母细胞减数分裂过程及染色体行为进行了观察研究,以探明同源四倍体矮牵生育性降低的细胞学原因.结果显示:花粉母细胞减数分裂过程与二倍体基本相同但有其特殊性,主要表现在:终变期染色体的构型复杂,有四价体、三价体和单价体;中期Ⅰ和中期Ⅱ有赤道板外染色体;后期Ⅰ和后期Ⅱ出现落后染色体、丢失染色体、染色体桥及不均等分裂的现象;四分体时期出现一分体、二分休、三分体以及含微核的异常三分体、四分体、多分体.花粉母细胞减数分裂过程中正常细胞平均达78.6%,异常细胞频率平均为21.4%.研究表明,同源四倍体矮牵生育性降低的细胞学原因是减数分裂过程中染色体行为异常.     

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。     

基于MSAP和SSR标记的高粱甲基化连锁群LGC、LGD的构建

以高粱(Sorghum bicolor(L.)Moench)品种‘B_2V_4’和‘1383-2’杂交获得的F_2群体为材料,通过SSR和MSAP标记检测高粱基因组差异,构建其甲基化遗传连锁群。结果显示,高粱甲基化连锁群LGC含有3个SSR标记和23个甲基化标记,覆盖高粱基因组44.3 cM;甲基化连锁群LGD含有4个SSR标记和8个甲基化标记,覆盖高粱基因组46.2 cM。LGC上甲基化位点仅来源于EcoRⅠ/MspⅠ酶切组合,而LGD上有来源于EcoRⅠ/MspⅠ和EcoRⅠ/HpaⅡ两种酶切组合的甲基化位点。在LGC连锁群Xtxp 69附近检测到一个密集的甲基化位点区域。研究结果表明MSAP标记可以快速检测植物基因组甲基化差异,适用于构建甲基化连锁群。     

辣椒属种间杂种F_1植株的细胞遗传学研究

本文分析了南京早椒（ＣａｐｓｉｃｕｍｆｒｕｔｅｓｃｅｎｓＬ．ｖａｒ．ＣｏｎｏｉｄｅｅｓＢａｉｌｅｙ）和Ｃ．ｃｈｉｎｅｎｓｅ二亲本及Ｆ１杂种花粉母细胞减数分裂终变期和中期Ｉ染色体配对的构型。结果表明：二亲本花粉母细胞减数分裂染色体配对均正常,为１２个二价体（双单倍体２ｎ＝２４）。Ｆ１杂种花粉母细胞减数分裂染色体配对很不规则。其平均频率分别为单价体０．０３６,二价体９．１８,三价体０．０３６,四价体０．８０,六价体０．３８。南京早椒和Ｃ．ｃｈｉｎｅｎｓｅ染色体间存在着相互易位,因此,二亲本间彼此有差异。单价体出现极少,这可断定南京早椒与Ｃ．ｃｈｉｎｅｎｓｅ染色体间是部分同源的,但染色体组结构上存在着差异。由于染色体结构上差异,所以Ｆ１杂种的育性较低。     

小麦-簇毛麦种质系‘山农030713＇的细胞学和SSR鉴定

利用形态学、细胞学以及SSR标记技术对从硬簇麦和Am3的杂种后代中选育的种质系‘山农030713＇进行了鉴定,结果表明：种质系‘山农030713＇大田生长整齐一致,农艺性状较好,且对白粉病免疫;其根尖细胞染色体数目为2n=42,花粉母细胞减数分裂中期Ⅰ（PMC M Ⅰ）染色体构型为2n=21Ⅱ;它与普通小麦的杂种F1PMC MⅠ多数细胞中形成21个二价体,且常有四价体出现,可能伴有染色体的结构变异;SSR分析证明‘山农030713＇基本染色体组成为AABBDD,引物Xgwm99-1A在‘山农030713＇中扩增出簇毛麦的特异带,表明‘山农030713＇中有来自于簇毛麦的遗传物质,此特异带可作为识别‘山农030713＇的SSR标记.综合形态学、细胞学和SSR分析结果推测,‘山农030713＇可能是一个小麦-簇毛麦易位系.     

利用不对称体细胞杂交向小麦转移高冰草染色体小片段

对小麦与高冰草不对称体细胞杂种F₅代株系Ⅱ-1-3和Ⅰ-1-9的染色体组成及异源染色体存在方式进行分析. 花粉母细胞观察统计发现两杂种中分别有84.69%和84.75%的细胞为20~21对二价体, 其中环状二价体分别占89.83%和89.57%, 表明两杂种的遗传组成基本稳定. RAPD分析证明两杂种含有双亲的DNA并发生了整合. GISH结果表明高冰草染色体以小片段的方式分布在杂种Ⅱ-1-3和Ⅰ-1-9的4~6个染色体上, 小片段位于小麦的近着丝粒位置及近端部. SSR分析结果表明高冰草DNA分布于两杂种株系的2A, 5B, 6B和2D染色体上, 与GISH分析的多位点小片段插入的结果相对应     

The genomic relationship between cultivated sorghum [Sorghum bicolor (L.) Moench] and Johnsongrass [S. halepense (L.) Pers.]: a re-evaluation

Summary The genomic relationship between cultivated sorghum [Sorghum bicolar (L.) Moench, race bicolor, De Wet, 2n=20] and Johnsongrass [S. halepense (L.) Pers., 2n=40] has been a subject of extensive studies. Nevertheless, there is no general consensus concerning the ploidy level and the number of genomes present in the two species. This research tested the validity of four major genomic models that have been proposed previously for the two species by studying chromosome behaviors in the parental species, 30-chromosome hybrids [sorghum, (2n=20) x Johnsongrass, (2n=40)], 40-chromosome hybrids [sorghum, (2n=40) x Johnsongrass, (2n=40)] and 60-chromosome amphiploids. Chromosome pairings of amphiploids are reported for the first time. Chromosomes of cultivated sorghums paired exclusively as 10 bivalents, whereas Johnsongrass had a maximum configuration of 5 ring quadrivalents with occasional hexavalents and octovalents. In contrast, 40-chromosome cultivated sorghum had up to 9 ring quadrivalents and 1 hexavalent. Pairing in the 30-chromosome hybrids showed a maximum of 10 trivalents, and that in the 40-chromosome hybrids exhibited 8 quadrivalents, 5 of which were rings, together with a few hexavalents. Amphiploid plants showed up to 3 ring hexavalents, 1 chain hexavalent and a chain of 12 chromosomes. The data suggest that cultivated sorghum is a tetraploid species with the genomic formula AAB1B1, and Johnsongrass is a segmental auto-allo-octoploid, AAAA B1B1B2B2. The model is further substantiated by chromosome pairing in amphiploid plants whose proposed genomic formula is AAAAAA B1B1B1B1 B2B2.Contribution no. 87-391-J from the Kansas Agriculatural Experiment Station     

Population genetics structure of glyphosate‐resistant Johnsongrass (Sorghum halepense L. Pers) does not support a single origin of the resistance

Single sequence repeats (SSR) developed for Sorghum bicolor were used to characterize the genetic distance of 46 different Sorghum halepense (Johnsongrass) accessions from Argentina some of which have evolved toward glyphosate resistance. Since Johnsongrass is an allotetraploid and only one subgenome is homologous to cultivated sorghum, some SSR loci amplified up to two alleles while others (presumably more conserved loci) amplified up to four alleles. Twelve SSR providing information of 24 loci representative of Johnsongrass genome were selected for genetic distance characterization. All of them were highly polymorphic, which was evidenced by the number of different alleles found in the samples studied, in some of them up to 20. UPGMA and Mantel analysis showed that Johnsongrass glyphosate‐resistant accessions that belong to different geographic regions do not share similar genetic backgrounds. In contrast, they show closer similarity to their neighboring susceptible counterparts. Discriminant Analysis of Principal Components using the clusters identified by K‐means support the lack of a clear pattern of association among samples and resistance status or province of origin. Consequently, these results do not support a single genetic origin of glyphosate resistance. Nucleotide sequencing of the 5‐enolpyruvylshikimate‐3‐phosphate synthase (EPSPS) encoding gene from glyphosate‐resistant and susceptible accessions collected from different geographic origins showed that none presented expected mutations in aminoacid positions 101 and 106 which are diagnostic of target‐site resistance mechanism.     

A Diploid,Interspecific, Fertile Hybrid from Cultivated Sorghum, <Emphasis Type="Italic">Sorghum Bicolor</Emphasis>, and the Common Johnsongrass Weed <Emphasis Type="Italic">Sorghum Halepense</Emphasis>

Valuable agronomic traits are often present but inaccessible in the wild relatives of cultivated crop species. Utilization of wild germplasm depends on the production of fertile interspecific hybrids. Several unsuccessful attempts have been made to hybridize cultivated sorghum with its wild relatives to broaden its genetic base and enhance agronomic value. The successful approach used in this study employed the nuclear male sterility gene ms3 to generate a diploid fertile hybrid between the diploid cultivated sorghum (Sorghum bicolor (L) Pers.) and its weedy tetraploid wild relative Johnsongrass (Sorghum halepense (L.) Pers.). Eight sorghum plants were selected from a Nebraska stiff stalk collection that contains the male sterility gene ms3 and were used as the female parent. About 36,000 florets of male sterile sorghum were pollinated with Johnsongrass pollen to produce an average of one well-developed and 180 severely shriveled seed/18,000 crosses. The well-developed seed gave rise to a self-fertile diploid, while none of the shriveled seed were able to germinate. The F₁ hybrid was confirmed by using cultivated sorghum SSR markers and was selfed to produce an F₂ population. A sub-sample of 96 segregating F₂ plants was examined with 36 sorghum polymorphic SSR markers. Thirty-four markers showed a normal 1:2:1 segregation ratio, evidence of normal recombination across the genome. Preliminary results showed that several desirable traits from Johnsongrass, including resistance to greenbug and chinch bug and adaptability to cold temperatures, were expressed in the resulting progenies. These observations suggest that speciation within the genus Sorghum, giving rise to widely divergent phenotypes, is effected largely by ploidy-maintained crossing barriers but apparently not by extensive genomic divergence.     

An SSR genetic map of Sorghum bicolor (L.) Moench and its comparison to a published genetic map.

Sorghum bicolor (L.) Moench is an important grain and forage crop grown worldwide. We developed a simple sequence repeat (SSR) linkage map for sorghum using 352 publicly available SSR primer pairs and a population of 277 F2 individuals derived from a cross between the Westland A line and PI 550610. A total of 132 SSR loci appeared polymorphic in the mapping population, and 118 SSRs were mapped to 16 linkage groups. These mapped SSR loci were distributed throughout 10 chromosomes of sorghum, and spanned a distance of 997.5 cM. More important, 38 new SSR loci were added to the sorghum genetic map in this study. The mapping result also showed that chromosomes SBI-01, SBI-02, SBI-05, and SBI-06 each had 1 linkage group; the other 6 chromosomes were composed of 2 linkage groups each. Except for 5 closely linked marker flips and 1 locus (Sb6_34), the marker order of this map was collinear to a published sorghum map, and the genetic distances of common marker intervals were similar, with a difference ratio 相似文献   

Wide-cross whole-genome radiation hybrid mapping of cotton (Gossypium hirsutum L.)

We report the development and characterization of a "wide-cross whole-genome radiation hybrid" (WWRH) panel from cotton (Gossypium hirsutum L.). Chromosomes were segmented by gamma-irradiation of G. hirsutum (n = 26) pollen, and segmented chromosomes were rescued after in vivo fertilization of G. barbadense egg cells (n = 26). A 5-krad gamma-ray WWRH mapping panel (N = 93) was constructed and genotyped at 102 SSR loci. SSR marker retention frequencies were higher than those for animal systems and marker retention patterns were informative. Using the program RHMAP, 52 of 102 SSR markers were mapped into 16 syntenic groups. Linkage group 9 (LG 9) SSR markers BNL0625 and BNL2805 had been colocalized by linkage analysis, but their order was resolved by differential retention among WWRH plants. Two linkage groups, LG 13 and LG 9, were combined into one syntenic group, and the chromosome 1 linkage group marker BNL4053 was reassigned to chromosome 9. Analyses of cytogenetic stocks supported synteny of LG 9 and LG 13 and localized them to the short arm of chromosome 17. They also supported reassignment of marker BNL4053 to the long arm of chromosome 9. A WWRH map of the syntenic group composed of linkage groups 9 and 13 was constructed by maximum-likelihood analysis under the general retention model. The results demonstrate not only the feasibility of WWRH panel construction and mapping, but also complementarity to traditional linkage mapping and cytogenetic methods.     

PEARL MILLET (PENNISETUM AMERICANUM (L.) LEEKE) AND GRAIN SORGHUM (SORGHUM BICOLOR (L.) MOENCH) ULTRASTRUCTURE

Ultrastructural features of pearl millet (Pennisetum americanum (L.) Leeke) and grain sorghum (Sorghum bicolor (L.) Moench) caryospses were investigated with thin sections of the dry, mature grain in the transmission electron microscope, and fractured kernels in the scanning electron microscope. The pericarp of those grains is comprised of three distinct layers: epicarp, mesocarp of parenchyma cells, and endocarp of compressed cross and tube cells. Mesocarp cells of grain sorghum contain starch granules embedded in a cytoplasmic matrix. The major constituent of sorghum and millet aleurone cells are aleurone grains (protein bodies) and lipid bodies. Subaleurone cells contain a much higher proportion of protein bodies than starch granules, and the protein bodies are structurally distinct from those in the aleurone. The germ scutellar ultrastructures of the two grains were similar; protein bodies, lipid bodies, epidermal cells and parenchyma cells of the germ are described.     

An integrated SSR and RFLP linkage map of Sorghum bicolor (L.) Moench.

We report the development, testing, and use (for genetic mapping) of a large number of polymerase chain reaction (PCR) primer sets that amplify DNA simple sequence repeat (SSR) loci of Sorghum bicolor (L.) Moench. Most of the primer sets were developed from clones isolated from two sorghum bacterial artificial chromosome (BAC) libraries and three enriched sorghum genomic-DNA (gDNA) libraries. A few were developed from sorghum DNA sequences present in public databases. The libraries were probed with radiolabeled di- and trinucleotide oligomers, the BAC libraries with four and six oligomers, respectively, and the enriched gDNA libraries with four and three oligomers, respectively. Both types of libraries were markedly enriched for SSRs relative to a size-fractionated gDNA library studied earlier. However, only 2% of the sequenced clones obtained from the size-fractionated gDNA library lacked a SSR, whereas 13% and 17% of the sequenced clones obtained from the BAC and enriched gDNA libraries, respectively, lacked a SSR. Primer sets were produced for 313 SSR loci. Two-hundred sixty-six (85%) of the loci were amplified and 165 (53%) of the loci were found to be polymorphic in a population composed of 18 diverse sorghum lines. (AG/TC)n and (AC/TG)n repeats comprised 91% of the dinucleotide SSRs and 52% of all of the SSRs at the polymorphic loci, whereas four types of repeats comprised 66% of the trinucleotide SSRs at the loci. Primer sequences are reported for the 165 polymorphic loci and for eight monomorphic loci that have a high degree of homology to genes. Also reported are the genetic map locations of 113 novel SSR loci (including four SSR-containing gene loci) and a linkage map composed of 147 SSR loci and 323 RFLP (restriction fragment length polymorphism) loci. The number of SSR loci per linkage group ranges from 8 to 30. The SSR loci are distributed relatively evenly throughout approximately 75% of the 1406-cM linkage map, but segments of five linkage groups comprising about 25% of the map either lack or contain few SSR loci. Mapping of SSR loci isolated from BAC clones located to these segments is likely to be the most efficient method for placing SSR loci in the segments.     

Variation in kafirin and alcohol-soluble glutelin chromatograms of sorghum inbred lines revealed by reversed-phase high-performance liquid chromatography

Summary Separations of kafirin and alcohol soluble glutelin proteins by reversed-phase high-performance liquid chromatography (RP-HPLC) from 7 inbreds and one hybrid of sorghum [Sorghum bicolor (L.) Moench] and one source of Johnsongrass [Sorghum halapense (L.) Pers.] were compared. Objectives were to assess the stability of protein profiles for seed sources produced at different locations and in different environments to examine the potential of RP-HPLC to provide genotypic profiles for sorghum. Analyses of variance data showed that levels of variation due to environments and locations were small; the majority of variation (93%) was among genotypes. Associations among inbreds revealed by multivariate and cluster analysis showed similarity with those that would be expected on the basis of pedigree. A chi-square analysis showed no deviation in the hybrid profile from the expected 21 ratio of peaks from the female and male inbred parents, respectively. Improvements in the ability to correctly assign common peaks are necessary before associations among numerous sorghum genotypes can be reliably demonstrated by analysis of data from reversed-phase high-performance liquid chromatography (RP-HPLC).     

Assessment of genetic diversity and relationship among a collection of US sweet sorghum germplasm by SSR markers

Sweet sorghum (Sorghum bicolor L.) is a type of cultivated sorghums and has been recognized widely as potential alternative source of bio-fuel because of its high fermentable sugar content in the stalk. A substantial variation of sugar content and related traits is known to exist in US sweet sorghum. The objectives of the study were to assess the genetic diversity and relationship among the US sweet sorghum cultivars and lines using SSR markers and to examine the genetic variability within sweet sorghum accessions for sugar content. Sixty-eight sweet sorghum and four grain sorghum cultivars and lines were genotyped with 41 SSR markers that generated 132 alleles with an average of 3.22 alleles per locus. Polymorphism information content (PIC) value, a measure of gene diversity, was 0.40 with a range of 0.03–0.87. The genetic similarity co-efficient was estimated based on the segregation of the 132 SSR alleles. Clustering analysis based on the genetic similarity (GS) grouped the 72 sorghum accessions into 10 distinct clusters. Grouping based on clustering analysis was in good agreement with available pedigree and genetic background information. The study has revealed the genetic relationship of cultivars with unknown parentage to those with known parentage. A number of diverse pairs of sweet sorghum accessions were identified which were polymorphic at many SSR loci and significantly different for sugar content as well. Information generated from this study can be used to select parents for hybrid development to maximize the sugar content and total biomass, and development of segregating populations to map genes controlling sugar content in sweet sorghum.     

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