数量性状的核质互作遗传效应分析:二倍体植株遗传模型

提出了能分析二倍体植株数量性状核质互作效应的遗传模型,该模型把控制数量性状总的遗传效应分为核效应、质效应和核质互作效应,以及它们分别与环境作用的效应。其中,核质互作效应可进一步分解为加性核质互作与显性核质互作。基于平衡与非平衡两种双列杂交试验设计,蒙特卡罗模拟结果表明:采用混合线性模型方法进行统计分析,可以有效地估计各项遗传效应值及其方差分量。此外,运用该模型对棉花的4个数量性状(单株铃数、衣分、2.5%跨长和麦克隆值)进行了遗传分析。     

利用双单倍体群体剖析水稻产量及其相关性状的遗传基础

主效QTL、上位性效应和它们与环境的互作(QE)都是数量性状的重要遗传因素。利用籼粳交珍汕97／武育粳2号F1植株上的花药进行组织培养得到的190个双单倍体群体和179个微卫星标记,通过两年两重复田间试验,采用混合线性模型方法分析了9个控制水稻产量及其相关性状的遗传效应,得到57个主效QTL,41对上位性互作,8对QTL与环境的互作和7对上位性效应与环境的互作。单个主效QTL解释这些性状1．3％～25．8％的表型方差。各性状QTL的累积表型贡献率达11．5％～66．8％。大多数性状之间具有显著的表型相关性,相关性较高的性状之间常具有较多共同或紧密连锁的QTL。结果表明,基因的多效性或紧密连锁可能是性状相关的重要遗传基础。     

基因型×环境互作效应对水稻穗部性状杂种优势的影响

采用包括基因型×环境互作效应的加性显性加性×加性上位性遗传模型,分析了籼粳杂交穗部性状杂种优势的两年资料．结果表明,７ 个穗部性状的杂种优势既由基因型控制也受环境（ 年份） 互作效应影响．主穗粒数、主穗长、一次和二次枝梗总长和着粒密度表现较强的正向群体平均优势,而一次和二次枝梗数则表现显著的负向优势．基因型×环境互作预测结果表明,主穗粒数、主穗长、一次枝梗总长在两年中的环境互作表现正效应,一次和二次枝梗数则为负效应,着粒密度和二次枝梗总长表现不同方向的效应．利用ＩＲ６６１５８３７（Ｐ１） 、ＩＲ６５６００８５（Ｐ２） 、明恢６３（Ｐ４） 和Ｒ６６９（Ｐ６） 进行杂交配组,其杂种后代易获得重穗型的穗部结构,可在超高产育种中加以利用．     

籼粳杂交稻穗部性状的遗传效应及其与环境互作

采用包括基因型与环境互作效应的加性显性加性×加性上位性遗传模型,分析了不同环境下籼粳杂交稻穗部性状的遗传特点.结果表明,除了主穗粒数的加性与环境互作和二次枝梗数的显性与环境互作不显著外,其他性状均存在显著和极显著的加性、显性、加性×加性上位性遗传效应及其与环境的互作效应,其中均以显性效应为主,显性与环境互作效应对枝梗性状的影响较为明显.遗传率分析表明,各性状的普通广义遗传率最大,互作遗传率也有一定作用.杂种优势预测表明,除了一次、二次枝梗数外,其他性状均表现正向的杂种优势,基因型与环境互作只影响杂种优势表达的程度,而不改变其方向.遗传效应预测值结果表明,IR6615837、IR6560085、明恢63和R6694个亲本可以明显改良杂交后代多数穗部性状,且环境影响程度较小,可作为优良亲本列于育种计划中.     

短季棉早熟性的遗传效应及其与环境互作研究

以熟期不同的9个棉花品种为亲本,按部分双列杂交配制46个组合的F1、F2,在3个不同生态环境条件下,研究了7个早熟相关性状的遗传效应及其与环境互作。结果表明:短季棉7个早熟相关性状的遗传均以加性效应为主,同时存在着显性效应,对于播种-现蕾、播种-开花和现蕾-开花还存在着上位性效应;短季棉各早熟性状的遗传效应与环境互作显著。生育期、播种-开花的狭义遗传率均较高,分别为66.1%和49.1%,且与环境互作效应较小,而果枝始节和播种-现蕾的遗传率最低,分别为19.8%和18.8%,且与环境互作达到极显著水平,现蕾-开花、开花-吐絮和株高这3个性状的遗传率及其与环境互作居中。由此说明:早熟性的遗传受环境影响较大,在生态条件差异较大的育种地点,以果枝始节和播种-现蕾作为早熟性指标进行异地选择是不可靠的,而以生育期、开花期为早熟性选择指标是比较可行的。     

五个箭筈豌豆品系基因型与环境互作效应及农艺性状稳定性

通过 2 a、4个区试点和 5种基因型及其互作效应研究 ,运用混合线性模型和 MINQU E(1)法 ,对箭豌豆包括株高在内的9个农艺性状可塑性进行评价 ,揭示了年份和区试点的生态环境效应、基因型与生态环境互作效应对各农艺性状的可塑性。结果表明 ,种子产量和千粒重等性状的基因型与生态环境互作效应达到了极显著水平 (p <0 .0 1和 p<0 .0 0 1)。生态环境分量 (年份、区试点、年份×区试点 )对各农艺性状的可塑性贡献较大 ,同时在不同生态环境间各农艺性状间差异达到了显著水平 (p <0 .0 5)。其中区试点分量对各农艺性状的可塑性贡献最大 ,各农艺性状在 4个区试点之间差异达到了显著水平 (p<0 .0 1) ,肃南和天祝的 2个区试点的牧草干重和种子产量等重要农艺性状的平均值显著大于另 2个区试点。各农艺性状在不同年份间差异达到了极显著水平 (p<0 .0 1) ,2 0 0 2年各农艺性状的平均值显著优于 2 0 0 1年。牧草干重和种子产量数量性状与气候因子的相关分析表明 ,5～ 8月份的月均温对牧草干重和种子产量的影响作用较大 ,较高的温度有利于牧草干重和种子产量的提高 ;7月份的降水量与牧草干重和种子产量存在一定程度的正相关关系。品系 2 556和 2 560在进行了基因型与环境互作效应稳定性评价后 ,4个区试点两年间都     

大豆小区产量及其相关性状QTL间的上位性和环境互作效应

以晋豆23栽培大豆(Glycine max)为母本、灰布支黑豆(ZDD2315,半野生大豆)为父本衍生出447个RIL群体,通过构建SSR遗传图谱及利用混合线性模型分析方法,对2年大豆小区产量及主要植物学性状进行QTL定位,并作加性效应、加性×加性上位互作效应及环境互作效应分析。结果显示,共检测到12个与小区产量、单株粒重、单株茎重、单株粒茎比、有效分枝、主茎节数、株高和结荚高度相关的QTL,分别位于A1、A2、H_1、I、J_2和M连锁群上。其中小区产量、株高、单株粒重、有效分枝和主茎节数均表现为遗传正效应,即增加其性状的等位基因来源于母本晋豆23。同时,检测到11对影响小区产量、单株粒重、单株茎重、株高和结荚高度的加性×加性上位互作效应及环境互作效应的QTL,发现22个QTL与环境存在互作。实验结果表明,上位效应和QE互作效应对大豆小区产量及主要农艺性状的遗传影响很大。进行大豆分子标记辅助育种时,既要考虑效应起主要作用的QTL,又要注重上位性QTL,这样有利于性状的稳定表达和遗传。     

籼粳杂交稻穗部性状的遗传效应及其与环境互作

采用包括基因型与环境互作效应的加性-显性-加性×加性上位性遗传模型,分析了不同环境下籼粳杂交稻穗部性状的遗传特点.结果表明,除了主穗粒数的加性与环境互作和二次枝梗数的显性与环境互作不显著外,其他性状均存在显著和极显著的加性、显性、加性×加性上位性遗传效应及其与环境的互作效应,其中均以显性效应为主,显性与环境互作效应对枝梗性状的影响较为明显.遗传率分析表明,各性状的普通广义遗传率最大,互作遗传率也有一定作用.杂种优势预测表明,除了一次、二次枝梗数外,其他性状均表现正向的杂种优势,基因型与环境互作只影响杂种优势表达的程度,而不改变其方向.遗传效应预测值结果表明,IR66158-37、IR65600-85、明恢63和R6694个亲本可以明显改良杂交后代多数穗部性状,且环境影响程度较小,可作为优良亲本列于育种计划中.     

第56位酪氨酸与肿瘤坏死因子细胞毒活性的关系*

选择肿瘤坏死因子分子中间部分保守区第56位酪氨酸,用寡核苷酸诱导的体外基因定点突变法,将其改变为谷氨酰胺。将突变的肿瘤坏死因子基因插入表达载体pBV220,所得表达产物的分子大小及表达量均无明显变化,但其细胞毒活性及比活性均下降数百倍,表弱该区是肿瘤坏死因子细胞因子细胞毒活性的必需区。     

肿瘤坏死因子第59位酪氨酸对其细胞毒活性的意义

肿瘤坏死因子(Tumor Necrosis Factor,TNF)的基因克隆表达成功后,已进行了广泛的实验室研究,并进入Ⅰ期及Ⅱ期临床试验。但关于肿瘤坏死因子结构与功能关系人们尚所知甚少,阻碍了对肿瘤坏死因子各种生物活性的机制研究,也影响了肿瘤坏死因子的临床应用。本研究为肿瘤坏死因子结构与功能关系研究的一部分。肿瘤坏死因子不仅有广泛的生物学活性,也可引起严重的毒副作用。本文用寡核苷酸诱导的体外基因定点突变法,选择位于肿瘤坏死因子分子中央保守区第59位酪氨酸,将其改变为门冬氨酸。所选用的琥珀突变选择性突变系统,造成肿瘤坏死因子基因突变,获得较高的突变效率(约80％)。DNA序列分析证实第59位酪氨酸密码子TAC突变为门冬氨酸密码子GAT。将突变体肿瘤坏死因子基因于同一原核表达载体进行表达,结果表明,突变后不影响表达量及表达产物分子量,但其细胞毒比活性下降724倍,表明Tyr59对于维持肿瘤坏死因子细胞毒活性非常重要。     

Inverse PCR-based site-directed mutagenesis of nucleotide sequences coding for carbohydrate-binding fragments of legume lectins

A new method of site-directed mutagenesis was developed to allow manipulation with extended plasmid-cloned gene fragments irrespective of their position and the presence of restriction sites. The method was used to obtain chimeric constructs encoding a Pisum sativum lectin with the native carbohydrate-binding region replaced by its counterpart from other legumes. The method can be used in plasmid construction to clone a coding gene fragment under the control of a promoter in a certain reading frame.     

陆地棉叶形及棉酚腺体变异的遗传研究

阔叶、有棉酚腺体陆地棉干种子经250Gy^60 Coγ射线辐照,在M1代筛选到鸡脚叶γ-1、γ-2和γ-5突变体。3株突变体自交后代（M2）中,阔叶：鸡脚叶：超鸡脚叶的分离比例符合1：2：1;γ—1和γ-2自交后代中的所有棉株均有棉酚腺体;γ-5自交后代中,有棉酚腺体株：无棉酚腺体株的分离比例符合3：1,阔叶、有棉酚腺体株：阔叶、无棉酚腺体株：鸡脚叶、有棉酚腺体株：鸡脚叶、无棉酚腺体株：超鸡脚叶、有棉酚腺体株：超鸡脚叶、无棉酚腺体株的分离比例符合3／16：1／16：6／16：2／16：3／16：1／16。该结果表明,3株突变体的叶形变异由1对不完全显性基因控制,γ-5的无棉酚腺体基因由1对隐性基因控制,且γ-5的叶形基因与棉酚腺体基因为独立遗传。通过对γ-5自交后代各种变异株的SSR标记研究表明,BNL3649、BNL2440度BNL1414为候选标记。为进一步对突变基因进行定位奠定了基础。     

Analysis of thermal adaptation in the HSL enzyme family

The recently solved three-dimensional (3D) structures of two thermostable members of the carboxylesterase/lipase HSL family, namely the Alicyclobacillus (formerly Bacillus) acidocaldarius and Archaeoglobus fulgidus carboxylesterases (EST2 and AFEST, respectively) were compared with that of the mesophilic homologous counterpart Brefeldine A esterase from Bacillus subtilis. Since the 3D homology models of other members of the HSL family were also available, we performed a structural alignment with all these sequences. The resulting alignment was used to assess the amino acid “traffic rule” in the HSL family. Quite surprisingly, the data were in very good agreement with those recently reported from two independent groups and based on the comparison of a huge number of homologous sequences from the genus Bacillus, Methanococcus and Deinococcus/Thermus. Taken as a whole, the data point to the statistical meaning of defined amino acid conversions going from psychrophilic to hyperthermophilic sequences. We identified and mapped several such changes onto the EST2 structure and observed that such mutations were localized mostly in loops regions or α-helices and were mostly excluded from the active site. A site-directed mutagenesis of two of the identified residues confirmed they were involved in thermal stability.     

短季棉早熟不早衰生化性状的遗传分析

用5个早熟不早衰的短季棉品种和5个早衰的短季棉品种进行双列杂交,并对亲本、F1和F2代于2001年和2002年田间试验研究与短季棉早熟不早衰有关的抗氧化系统保护酶(SOD、POD和CAT)、叶绿素及激素(生长素和脱落酸)的遗传特性。结果表明：抗氧化系统保护酶CAT、POD和SOD存在着不同的遗传特性,CAT酶以加性上位性效应为主,其次为显性效应;POD酶以加性效应为主,其次为加性上位性效应;SOD酶活以显性效应为主,其次为加性上位性效应;IAA以显性效应为主,其次为加性效应;ABA以加性效应为主,其次为显性效应。且这些生化性状的遗传率较高,在后代能稳定遗传;同时棉株在不同发育时期体内生化性状表达不同,在花铃期CAT、POD和SOD酶以显性效应为主,其次为加性上位性效应,加性效应表达量很小;棉株体内生化性状的表达也是相互联系、相互制约,CAT酶与POD酶存在着遗传和表型负相关,与SOD酶存在着遗传和表型正相关;POD酶与SOD酶存在着遗传和表型负相关;抗氧化系统保护酶与激素之间存在着复杂的遗传关系。因此,研究生化性状的遗传特性和表达特征,为选育短季棉早熟不早衰新品种和生化性状的QTLs定位提供理论依据。     

基于SSR标记的陆地棉早熟相关种质遗传多样性分析

丰富的遗传变异对于提高作物的环境适应性和遗传改良进度至关重要。为了解我国早熟陆地棉种质资源遗传多样性,本研究利用136对SSR引物对186份陆地棉材料(96份早熟陆地棉材料和90份中、晚熟陆地棉材料)进行了遗传多样性分析,共检测到等位基因变异355个,平均2.61个。在早熟棉材料中,有134对多态性SSR引物扩增出341个条带,平均2.54个;中、晚熟材料中有133对多态性SSR引物,扩增出345个条带,平均2.59个。早熟棉材料的位点多态性信息含量(PIC)、有效等位基因数(Ne)、基因型多样性(H')分别为0.684、3.994和1.361;中、晚熟棉材料的PIC、Ne、H'分别为0.668、3.852和1.343。早熟棉材料和中、晚熟棉材料的Jaccard相似性系数分别在0.349~0.935和0.270~0.907之间,平均为0.635、0.666。遗传相似性系数总体平均值接近,但早熟棉变化范围较中、晚熟棉小。用类平均法(UPGMA)进行聚类可将186份材料分成2个类群。总体上来看,供试材料遗传相似性系数较高,说明我国陆地棉早熟相关种质遗传基础狭窄。本研究结果为早熟棉育种亲本选配,早熟棉种质创新提供依据。     

Functional Analysis of the Thioredoxin Domain in Porphyromonas gingivalis HBP35

Periodontitis is one of the most common oral diseases in humans. This caused by infection by the oral bacterium Porphyromonas gingivalis. Our strategy to prevent this infection is to establish a passive immunization system in which endogenous antibodies can be applied directly to neutralize virulent factors associated with this bacterium. We focused our attention on the P. gingivalis 35 kDa surface protein, or HBP35, since this protein is involved not only in the coaggregation with oral miroflora but also in hemin binding. In addition, nucleotide sequencing of the gene, hbp35, coding for this protein revealed the presence of a catalytic center for thioredoxin, and we further attempted to characterized the protein by amino acid substitution. A total of four Cys residues were substituted for Ser residues by combining the simple method for site-directed mutagenesis and the heterodimer system, an approach designed to construct chimeric plasmids readily. Native and mutagenized hbp35 were introduced into the Eschericha coli dsbA mutant strain, JCB 572, defective in both alkaline phosphatase and motile activities due to inefficient disulfide bond formation. Transformant harboring the native hbp35 could complement the dsbA mutation, suggesting a role of disulfide bond formation of this protein in P. gingivalis cells. Possible roles of the Cys residues in complementation are discussed.     

棉花WRKY基因GhWRKY25的克隆和表达分析

棉花是一种重要的经济作物,在国民生产中起着重要的作用。然而干旱盐碱等不良环境严重地制约着棉花的种植区域及其产量和品质。近年来,利用分子育种改良棉花对这些逆境的耐性的研究成为热点。本文从棉花中克隆了一个对干旱和盐碱具有应答的WRKY基因,定名为GhWRKY25。该基因编码的蛋白包含两个典型的WRKY结构域,每个WRKY结构域的C端含有一个C2H2类型的锌指蛋白结构,进化树分析表明其属于Ⅰ类WRKY蛋白,与可可的WRKY3同源性最高。利用qPCR分析发现该基因在棉花里为组成型表达,但在花中表达量较高。GhWRKY25在施用SA、JA、ABA、GA 中表达表现为上调。干旱和NaCl处理时发现GhWRKY25基因表达均显著上调。通过上述结果可以推测GhWRKY25在棉花对于逆境的适应中具有重要的作用,为该基因功能的进一步研究提供基础。     

Interactions between Multiple Phosphorylation Sites in the Inactivation Particle of a K+ Channel : Insights into the Molecular Mechanism of Protein Kinase C Action

Protein kinase C inhibits inactivation gating of Kv3.4 K⁺ channels, and at least two NH₂-terminal serines (S15 and S21) appeared involved in this interaction (Covarrubias et al. 1994. Neuron. 13:1403–1412). Here we have investigated the molecular mechanism of this regulatory process. Site-directed mutagenesis (serine → alanine) revealed two additional sites at S8 and S9. The mutation S9A inhibited the action of PKC by ∼85%, whereas S8A, S15A, and S21A exhibited smaller reductions (41, 35, and 50%, respectively). In spite of the relatively large effects of individual S → A mutations, simultaneous mutation of the four sites was necessary to completely abolish inhibition of inactivation by PKC. Accordingly, a peptide corresponding to the inactivation domain of Kv3.4 was phosphorylated by specific PKC isoforms, but the mutant peptide (S[8,9,15,21]A) was not. Substitutions of negatively charged aspartate (D) for serine at positions 8, 9, 15, and 21 closely mimicked the effect of phosphorylation on channel inactivation. S → D mutations slowed the rate of inactivation and accelerated the rate of recovery from inactivation. Thus, the negative charge of the phosphoserines is an important incentive to inhibit inactivation. Consistent with this interpretation, the effects of S8D and S8E (E = Glu) were very similar, yet S8N (N = Asn) had little effect on the onset of inactivation but accelerated the recovery from inactivation. Interestingly, the effects of single S → D mutations were unequal and the effects of combined mutations were greater than expected assuming a simple additive effect of the free energies that the single mutations contribute to impair inactivation. These observations demonstrate that the inactivation particle of Kv3.4 does not behave as a point charge and suggest that the NH₂-terminal phosphoserines interact in a cooperative manner to disrupt inactivation. Inspection of the tertiary structure of the inactivation domain of Kv3.4 revealed the topography of the phosphorylation sites and possible interactions that can explain the action of PKC on inactivation gating.     

Genetic Diversity of Source Germplasm of Upland Cotton in China as Determined by SSR Marker Analysis

The genetic diversity of 43 sources of Upland cotton germplasm with different parental origins, breeding periods, and ecological growing areas in China were studied on the basis of simple sequence repeat (SSR) markers. A total of 130 gene alleles with 80% polymorphism were detected from 36 SSR primers. The number of alleles per primer ranged from two to eight with an average of 3.6. The polymorphism information content (PIC) range was 0.278-0.865, with an average of 0.62. The average genotype diversity index (H') was 1.102, the highest was 2.039 and the lowest was 0.451. The average coefficient of the genetic similarity of SSR markers among source germplasm was 0.610, ranging from 0.409 to 0.865. These indicated that the genetic diversity at the genomic level of the selected source germplasm was rich, and was representative of the diversity of the germplasms, in general. The diversity at the genome level of the base germplasm from the second and third breeding periods was decreased compared to that of the first period, indicating that the cotton genetic background in China became narrow gradually. The diversity of SSR markers among the base germplasm from early maturity cotton growing areas in the north was higher than those from the Huanghe and Yangtze growing areas. The molecular marker genetic similarity index of the domestic varieties was higher than that in the introduced varieties, which indicates that the genetic diversity in domestic cultivars was lower than that in the introduced varieties. This study gives an overview of the genetic diversity of the cotton germplasm base in China, and provides a guide for breeders to develop new cultivars efficiently.