配子体自交不亲和植物花粉S基因研究进展

配子体自交不亲和植物的自交不亲和性是由雌蕊自交不亲和因子和花粉自交不亲和因子相互作用的结果。目前已经分离和鉴定了雌蕊自交不亲和基因及其表达产物。最近从金鱼草、Prumusdulcis、梅等植物中分离的F-box基因,它具有花粉S基因特点,即在花药、成熟的花粉和花粉管中特异表达;在基因位置上,与S-RNase基因紧密连锁;不同物种或同一物种不同品种F-box基因间核苷酸和氨基酸序列上存在高度多态性。通过分子生物学方法和杂交授粉试验证明所分离的F-box基因是花粉自交不亲和基因,但目前尚未分离出该类基因相应的表达蛋白。主要综述了配子体自交不亲和植物花粉自交不亲和基因的发现、基因的结构、雌蕊自交不亲和因子和花粉自交不亲和因子相互作用的模型。     

油茶自、异交雌蕊CAT活性及相关基因克隆表达分析

油茶(Camellia oleifera Abel.)具有后期自交不亲和(LSI)特性,严重阻碍了油茶杂交育种的亲本选择和品种配置。为研究过氧化氢酶(Catalase, CAT)在油茶自交不亲和中的作用,本文对油茶自、异交授粉后雌蕊中的CAT活性进行了测定,并克隆得到3个CAT基因,命名为CoCAT1～CoCAT3。结果显示,其CDS分别为1479、1287和1479 bp,编码492、428和492个氨基酸。多序列比对结果表明,其编码产物在C末端均含有典型的三肽序列。系统进化分析发现,CoCAT2、CoCAT3与NtCAT3亲缘关系最近,CoCAT1与CsCAT1亲缘关系最近。基因表达模式分析结果显示,3个基因在授粉72 h后的自交雌蕊中表达量最大,在授粉36、84 h自交雌蕊中的表达量显著高于异交。自、异花授粉均引起油茶雌蕊内CAT活性的上升,但幅度不同,推测油茶CAT可能参与油茶自交不亲和的响应机制。     

植物自交不亲和基因研究进展

自交不亲和性的研究是植物生殖生物学和分子生物学研究的热点之一,对自交不亲和基因和蛋白质的深入研究是解析自交不亲和性机理的关键.对控制孢子体自交不亲和性和配子体自交不亲和性的S基因及其蛋白质产物的分子生物学研究进展进行了综述.孢子体自交不亲和性植物S位点上至少存在3个基因,即SLG、SRK和SCR基因.其中SLG、SRK基因控制雌蕊自交不亲和性,而SCR控制花粉自交不亲和性.配子体自交不亲和植物雌蕊S基因产物为S-RNase,具有核酸酶活性;配子体自交不亲和植物花粉S基因产物尚未找到.     

花粉——雌蕊相互作用的分子基础

显花植物授粉过程包含了花粉与雌蕊一系列复杂的细胞间相互作用。花粉在柱头上必须发生粘附与水合作用后才能萌发,花粉胞被蛋白可能在粘附机制中起主要作用,而水孔蛋白调节了花粉的水合过程;花粉管在花柱引导组织中定向生长,受引导组织胞间质、向化性物质及细胞粘附机制等因素的影响,也与花粉受体及子房有关。在植物的自交不亲和性反应中,配子体型自交不亲和性反应主要是由自交不亲和基因蛋白产物降解花粉RNA或以Ca^2 介导的信号级联反应实现的;孢子体型自交不亲和性反应则依赖于干性柱头以及雌蕊的S受体激酶及S位点糖蛋白与花粉S基因配体之间的相互作用。     

柠檬UBE2I同源基因的克隆及表达模式分析

前期从香水柠檬自交不亲和花的转录组数据中筛选到一个上调表达的泛素结合酶UBE2I同源基因片段,为了弄清该基因的特性,开展了本研究。以香水柠檬(自交不亲和)和白花柠檬(自交亲和)为试材,根据从香水柠檬花转录组中获得的UBE2I基因片段设计引物,采用同源克隆方法获得了香水柠檬和白花柠檬这2个品种中UBE2I同源基因的ORF全长和DNA全长,对其序列进行生物信息学分析,再对其表达模式进行探讨。结果显示UBE2I基因在香水柠檬和白花柠檬中的ORF长度均为483 bp,二个品种ORF全长仅有一个核苷酸的差异,编码160个氨基酸,但其氨基酸序列无差异;DNA全长为2 267 bp,拥有有5个外显子和4个内含子,在第2个和第4个内含子序列中存在2个类似于启动子(TATAA)框的结构。UBE2I泛素连接酶蛋白理论等电点为7.64,分子量为17 981.474 44 Da,三级结构中有4个α螺旋、7个β折叠和10个无规卷曲,不是分泌蛋白,没有跨膜结构,肽链呈现疏水性。时空表达分析表明,UBE2I基因在香水柠檬的不同组织中均表达,但在不同组织中的表达存在差异,其中在花粉中的表达量明显高于其它组织,UBE2I在自交(香水柠檬♀×香水柠檬♂)后柱头中的表达量均比(香水柠檬♀×白花柠檬♂)杂交后柱头中的表达量高。本研究为进一步探索发现自交不亲和过程的泛素结合酶UBE2基因奠定基础。     

欧报春二型花柱的繁育特性研究

二型花柱植物通常具有自交不亲和性,在不同植物中所表现的可育性不同。欧报春(Primula vulgaris)是典型的二型花柱植物,为了探究欧报春的繁育特性,通过温室栽培,对欧报春的长花柱和短花柱的花部特征、花粉和花柱形态、花粉活力、柱头可授性、花粉/胚珠比、杂交指数、杂交亲和性、花粉管观察进行研究。结果表明,(1)长花柱和短花柱的花冠直径、花冠筒长度、柱头高度、花药高度和花筒中部直径均表现出两型性;(2)花粉和柱头观察发现花粉极轴长、花粉赤道轴长、花柱直径、柱头乳突细胞和花粉数量均有差异;(3)长花柱的花粉/胚珠比为384.20,短花柱为369.70,属于兼性异交类型;长花柱和短花柱的花粉活力和柱头可授性能在较长时间内维持较高活力;(4)长花柱杂交指数值为5,短花柱为4,表明繁育系统类型为异交,部分自交亲和,需要传粉者;6种授粉组合均能结实,异型花间授粉的结实数明显高于同型自花授粉和异株同型授粉;短花柱为母本的异型花间授粉组合亲和性优于其他组合;欧报春存在自交不亲和性,长花柱的自交不亲和性低于短花柱。     

油茶PODs基因鉴定及在自交和异交下的表达分析

油茶（Camellia oleifera Abel.）具有自交不亲和性（SI,self-incompatibility）,自然座果率低,这严重影响油茶产量,制约了油茶产业的发展。为研究过氧化物酶（POD,peroxidase）在油茶自交不亲和反应中的作用,本研究通过逆转录克隆技术从油茶中克隆出4条POD基因,分别命名为CoPOD1/2/3/4。其基因编码区长度分别为1086、1011、1020和1218 bp,编码361、336、339和405个无跨膜结构、有信号肽的蛋白质。同源序列比对显示CoPOD1/2/3/4蛋白质之间的同源性较低,但均有过氧化物酶活性位点和过氧化酶近端血红素配体序列;系统进化树显示CoPOD1/2/3/4蛋白质与茶树（Camellia sinensis(L.)O.Ktze.）POD蛋白质的亲缘关系最近。实时荧光定量结果显示,油茶PODs基因在自交授粉后24～48 h范围内呈现显著上调后又下降的趋势,在自交授粉36 h雌蕊中CoPOD1/3/4的表达量高于异交。自交授粉24～72 h油茶雌蕊内POD酶活性高于异交并在36 h达到最高值,异交授粉雌蕊内POD活性前期...     

高等植物自交不亲和性的分子生物学

自交不亲和性是大多数高等植物防止近亲繁殖的一种遗传屏障。它涉及受精时雄配子（花粉）和雌蕊之间的相互作用。目前,已经分离获得了编码控制雌蕊自交不亲和性的Ｓ基因。在孢子体型自交不亲和的芸苔属中,雌蕊Ｓ基因编码Ｓ位点糖蛋白（ＳＬＧ）和Ｓ受体激酶（ＳＲＫ）。它们可能与磷酸化和去磷酸化参与了的某种信号传递有关,最后导致自交花粉生长的抑制。在配子分配体型自交不亲和的茄科中,雌蕊Ｓ位点糖蛋白为一种核糖核酸酶,称     

大白菜BrCML49基因的分离鉴定及表达特征分析

类钙调蛋白(Calmodulin like protein,CML)是植物细胞中一类非常重要的钙感受蛋白,且CML蛋白在花粉萌发和花粉管生长中起到重要作用。该研究以大白菜自交不亲和系‘91 125’自花授粉后柱头为材料,克隆获得大白菜BrCML49基因cDNA全长序列1 046 bp,包括960 bp开放阅读框,编码319个氨基酸,包含2个EF hand保守结构域。序列分析发现,BrCML49蛋白为稳定的亲水蛋白,没有跨膜区。实时荧光定量(qRT PCR)表达分析显示,BrCML49基因的表达量在‘91 125’的花药中最高,其次是全花和花瓣,在其他组织中表达量较低;BrCML49基因在大白菜自交亲和系‘14S23’的全花和柱头中表达量较高,在其他组织中表达量较低。亚细胞定位分析发现BrCML49主要定位于烟草叶片的细胞膜和细胞核中。该研究结果可为进一步研究大白菜BrCML49基因的功能奠定基础,并为阐明大白菜BrCML49基因参与自交不亲和花粉萌发及花粉管生长的信号机制提供理论依据。     

植物的生殖讲座（五）：被子植物的自交不亲和性

自交不亲和性广泛存在于被子植物中,同形花与异型花均存在自交不亲和性。受精的障碍可发生在花粉萌发、花粉管进入柱头、花粉管在花柱中生长及进入胚囊中等不同阶段和部位。不亲和性由孢子体系统或配子体系统控制。用转基因技术研究发现甘蓝的ＳＬＧ启动子能控制配子体型和孢子体型的表达。配子体自交不亲和的Ｓ基因产物具有核酸酶的活性,能选择性地破坏不亲和花粉管的ＲＮＡ。本文简介了克服自交不亲和性的方法及自支不亲和性的利用。     

Ester production in E. coli and C. acetobutylicum

Genetic engineering has improved the product yield of a variety of compounds by overexpressing, inactivating, or introducing new genes in microbial systems. The production of flavor-enhancing ester compounds is an emerging area of heterologous gene expression for desired product yield in Escherichia coli. Isoamyl acetate, butyl acetate, ethyl acetate, and butyl butyrate are reported here to be produced by expressing Saccharomyces cerevisiae genes ATF1 or ATF2 and the strawberry gene SAAT in E. coli when the appropriate substrates are provided. Increasing the concentration of alcohol added to the reaction generally resulted in increased ester production. ATF1 expression was found to produce more isoamyl acetate and butyl acetate than ATF2 expression or SAAT expression in the strains and culture conditions examined. Additionally, SAAT expression resulted in greater isoamyl acetate and butyl acetate production than ATF2 expression. Butyl butyrate is produced by cell-free extracts of E. coli harboring SAAT but not ATF1 or ATF2.     

Chemical composition, in situ ruminal degradability and post-ruminal disappearance of dry matter and crude protein from the halophytic plants Kochia scoparia, Atriplex dimorphostegia, Suaeda arcuata and Gamanthus gamacarpus

Samples of Kochia (K. scoparia), Atriplex (A. dimorphostegia), Suaeda (S. arcuata) and Gamanthus (G. gamacarpus) were collected and analyzed for chemical composition including crude protein (CP), ether extract (EE), ash, neutral detergent fiber (NDFom), acid detergent fiber (ADFom), non-protein N (NPN), Ca, P, Na, K, Cl, Mg, Fe, Cu and Se. In addition, in situ ruminal degradability and post-ruminal disappearance of dry matter (DM) and CP of the samples using a mobile bag technique were determined. Results indicate that the chemical composition of Kochia and Atriplex was notably different from those of Suaeda and Gamanthus. All of these halophytic plants had high concentrations of Na, K, Cl, Cu and Se, and low levels of Ca, P and Mg. The rapidly degradable fractions of DM and CP (g/g) of Kochia (0.31 and 0.35, respectively) and Atriplex (0.39 and 0.50, respectively) were lower than for Suaeda (0.53 and 0.55, respectively) and Gamanthus (0.56 and 0.66, respectively). Ruminal DM and CP disappearance of Kochia (444 and 517 g/kg, respectively) and Atriplex (472 and 529 g/kg, respectively) were lower (P<0.05) than those of Suaeda (553 and 577 g/kg, respectively) and Gamanthus (663 and 677 g/kg, respectively) (P<0.05) using the mobile bag technique. Suaeda had the lowest (P<0.05) NDFom and ADFom disappearance (214 and 232 g/kg, respectively) in the rumen. Kochia scoparia and Atriplex dimorphostegia have more beneficial chemical nutritive components and digestible values versus Suaeda arcuata and Gamanthus gamacarpus.     

Flavonoids of Astilbe and Rodgersia compared to Aruncus

The flavonoid profiles of Astilbe (four taxa studied) and Rodgersia (two taxa studied) are based on simple flavonol glycosides. Astilbe has 3-O-mono-, 3-O-di-, and 3-O-triglycosides of kaempferol, quercetin, and myricetin, while Rodgersia has only mono- and diglycosides of kaempferol and quercetin. Astilbe×arendsii was also shown to accumulate dihydrochalcone glycosides. The flavonoid profile of Rodgersia is the simplest recorded so far in the herbaceous Saxifragaceae. The flavonoids of two species of Aruncus were shown to be based upon kaempferol and quercetin 3-O-mono- and 3-O-diglycosides. One of the species also exhibited an eriodictyol glycoside. The triglycoside differences were not considered important, but the differences in myricetin occurrences were taken as evidence against derivation of Saxifragaceae from an Aruncus-like ancestor. Should such an event be proposed, however, serious consideration would have to be given to the current pattern of myricetin occurrence in the two families.     

Multiple invasions of Gypsy and Micropia retroelements in genus Zaprionus and melanogaster subgroup of the genus Drosophila

Background  

The Zaprionus genus shares evolutionary features with the melanogaster subgroup, such as space and time of origin. Although little information about the transposable element content in the Zaprionus genus had been accumulated, some of their elements appear to be more closely related with those of the melanogaster subgroup, indicating that these two groups of species were involved in horizontal transfer events during their evolution. Among these elements, the Gypsy and the Micropia retroelements were chosen for screening in seven species of the two Zaprionus subgenera, Anaprionus and Zaprionus.     

百合LbCAT和LbGPX基因的克隆与表达分析

以切花百合(Lilium brownii var. viridulum)‘卡瓦纳’cDNA为模板,克隆了过氧化氢酶(LbCAT)和谷胱甘肽过氧化物酶(LbGPX)基因。序列分析表明,这2个基因分别包含1 479 bp和519 bp的开放阅读框(ORF),编码492个和172个氨基酸。进化分析结果表明,LbCAT蛋白与岷江百合CAT蛋白的氨基酸序列相似性最高(99.19%),且亲缘关系最近;LbGPX蛋白与油棕GPX蛋白的氨基酸序列相似性最高(78.61%),亲缘关系最近。qRT PCR结果显示,LbCAT和LbGPX在百合根、鳞茎、叶和花中都有表达。LbCAT在叶中表达量最高,LbGPX在花中表达量最高。这2个基因在百合花蕾的生长发育过程中均有表达,且表达量逐渐增加;在PEG处理后2个基因的转录水平升高,但独角金内酯(SLs)处理却显著降低了这2个基因的转录水平;该结果为百合抗逆性机理研究以及抗逆育种奠定了基础。     

Quinua and Relatives (Chenopodium sect.Chenopodium subsect.Celluloid)

Traditionally viewed as an Andean grain crop,Chenopodium quinoa Willd. includes domesticated populations that are not Andean, and Andean populations that are not domesticated. Comparative analysis of leaf morphology and allozyme frequencies have demonstrated that Andean populations, both domesticated(quinua) and free-living(ajara), represent an exceptionally homogeneous unit that is well differentiated from allied domesticates of coastal Chile(quingua) and freeliving populations of the Argentine lowlands(C. hircinum). This pattern of relationships indicates that Andean populations represent a monophyletic crop/weed system that has possibly developed through cyclic differentiation (natural vs. human selection) and introgressive hybridization. Relative levels of variation suggest that this complex originated in the southern Andes, possibly from wild types allied withC. hircinum, with subsequent dispersal north to Colombia and south to the Chilean coast. Coastal populations were apparently isolated from post-dispersal differentiation and homogenization that occurred in the Andes. Other data point toward a center of origin in the northern Andes with secondary centers of genetic diversity subsequently developing in the southern Andes and the plains of Argentina. Comparative linkage of South American taxa, all tetraploid, with North American tetraploids of the subsection will eventually clarify this problem. While the possibility of a direct phyletic connection betweenC. quinoa and the Mexican domesticate(C. berlandieri subsp. nuttalliae,) cannot be excluded, available evidence indicates that the latter represents an autonomous lineage that is associated with the basal tetraploid, C. b. subsp.berlandieri, through var.sinuatum, whereas South American taxa show possible affinities to either var. zschackei or var.berlandieri. An extinct domesticate of eastern North America,C. b. subsp.jonesianum, represents either another instance of independent domestication, possibly from subsp. b. var.zschackei, or a northeastern outlier of subsp.nuttalliae.     

Cloning and characterization of femA and femB from Staphylococcus epidermidis

A DNA fragment was identified and cloned from Staphylococcus epidermidis (Se) using femA from S. aureus (Sa) as a heterologous hybridization probe. DNA sequence analysis of a portion of this clone revealed two complete ORFs highly related to femA and femB of Sa. The genomic arrangement of the Se femA/B complex was nearly identical to that observed in Sa. Intra- and interspecies relatedness of these genes and conservation of genomic organization were consistent with gene duplication of one of these genes in an ancestral organism. Recombinant FEMA, produced in Escherichia coli (Ec), was purified to near homogeneity. Identity of the purified protein was verified by N-terminal amino acid (aa) sequence analysis.