两种广西特有报春苣苔属(苦苣苔科)植物传粉生物学研究

通过野外观察和室内试验,对两种广西特有的桂林小花苣苔和阳朔小花苣苔的传粉生物学进行比较研究。结果表明:两种报春苣苔属植物花期从7月底至8月初;单花花期内花粉具有较高的活性,最高可达92.6%和94.5%;柱头可授性最高时可达90%和95%;花粉/胚珠比率(P/O)分别为110.28±17.45和229.65±18.00;柱头与花药之间存在着空间隔离,可防止自花授粉;在单花花冠裂片张开后第3天,花药开始散粉,但雌蕊在花朵刚开始开放时已伸长,待第3天时柱头已伸长到位于花冠筒口部,高于花药,便于接受异花花粉;该两种植物均不存在无融合生殖现象;两种报春巨苔属植物均高度自交亲和,但很难发生自发的自花授粉,必须依靠外力,因此传粉媒介对于结实率有重要影响,自然条件下基本不发生自花授粉;在自然状态下结实率明显低于两种人工授粉的结实率;小蜂和淡脉隧蜂是目前已知仅见的两种传粉者。     

Morphological, chemical and genetic differentiation of two subspecies of Cistus creticus L. (C. creticus subsp. eriocephalus and C. creticus subsp. corsicus)

Cistus creticus L., an aromatic species from the Mediterranean area, contains various diterpenes bearing the labdane skeleton. The production of essential oil from this species has potential economic value, but so far, it has not been optimized. In order to contribute to a better knowledge of this species and to its differentiation, the morphological characters, volatile chemical composition and genetic data of two subspecies (C. creticus subsp. eriocephalus and C. creticus subsp. corsicus) were investigated. The leaf trichomes were studied using scanning electron microscopy. The chemical composition of Corsican essential oil (C. creticus subsp. corsicus) has been reported using GC, GC/MS and ¹³C NMR; the main constituents were oxygenated labdane diterpenes (33.9%) such as 13-epi-manoyl oxide (18.5%). Using plant material (54 samples) collected from 18 geographically distinct areas of the islands of Corsica and Sardinia, the basis of variation in the headspace solid-phase microextraction volatile fraction and an inter-simple sequence repeat genetic analysis were also examined. It was shown that the two subspecies of C. creticus differed in morphology, essential oil production, volatile fraction composition and genetic data.     

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.     

Description of Tropicibacter mediterraneus sp. nov. and Tropicibacter litoreus sp. nov.

Four strains (M15∅_3, M17^T, M49 and R37^T) were isolated from Mediterranean seawater at Malvarrosa beach, Valencia, Spain. Together with an older preserved isolate (strain 2OM6) from cultured oysters at Vinaroz, Castellón, Spain, the strains were thoroughly characterized in a polyphasic study and were placed phylogenetically within the Roseobacter clade in the family Rhodobacteraceae. Highest 16S rRNA sequence similarities of the five strains to the types of any established species corresponded to Tropicibacter multivorans (95.8–96.4%), Phaeobacter inhibens (95.9–96.3%) and Phaeobacter gallaeciensis (95.9–96.2%). On the other hand, whole genome (ANI) and protein fingerprinting (MALDI-TOF) data proved: (i) non clonality among the strains, and (ii) the existence of two genospecies, one consisting of strains M15∅_3, M17^T, M49 and 2OM6 and another one consisting of strain R37^T. Phenotypic traits determined allow differentiating both genospecies from each other and from closely related taxa. In view of all data collected we propose to accommodate these isolates in two species as members of the genus Tropicibacter, Tropicibacter mediterraneus sp. nov. (type strain M17^T = CECT 7615^T = KCTC 23058^T) and Tropicibacter litoreus sp. nov. (type strain R37^T = CECT 7639^T = KCTC 23353^T).     

Growth and isoflavonoid accumulation of Pueraria candollei var. candollei and P. candollei var. mirifica cell suspension cultures

We established cell suspension cultures derived from leaf, stem, and root calli of Pueraria candollei var. candollei and P. candollei var. mirifica using liquid Murashige and Skoog (MS) medium supplemented with 0.56 μM 6-benzyladenine (BA) and 4.52 μM 2,4-dichlorophenoxyacetic acid (2,4-D). Growth of the cell suspension cultures progressed to the stationary phase within 15–24 days. Methanolic extracts of cell suspension cultures of both varieties of P. candollei were analyzed using a validated HPLC protocol. All cell lines derived from leaf, stem, and root explants produced four major isoflavonoids: daidzein, daidzin, genistein, and genistin; these isoflavonoids were detected only in the roots of intact plants. Furthermore, the isoflavonoid contents of the cell suspension cultures were higher than those of intact plants. Thus, cell suspension culture of both varieties of P. candollei may be an effective tool for isoflavonoid production.     

青稞HvBAS1基因的克隆及表达分析

细胞色素P450单加氧酶(CYP450)是参与植物代谢的最大酶家族,其中CYP734A亚家族成员广泛参与植物激素油菜素类固醇(BRs)的失活。该研究以青稞农家品种‘肚里黄’幼苗为实验材料,通过人工合成激素24 表油菜素内酯(24 eBL)和BRs合成抑制剂油菜素唑(BRZ)处理,分析BRs对青藏高原特色作物青稞(Hordeum vulgare L. var. nudum Hook. f.)的影响;采用RT PCR技术从青稞中克隆HvBAS1基因,并运用实时定量PCR检测其表达特征,为深入分析HvBAS1基因的功能奠定研究基础。结果显示：(1)24 eBL能够显著促进青稞幼苗的生长,而BRZ处理后幼苗长势明显减缓。(2)从青稞中成功克隆到2个与拟南芥BRs失活基因BAS1高度同源的CYP734A亚家族基因,即HvBAS1 1和HvBAS1 2;HvBAS1 1开放阅读框全长1 629 bp,编码542个氨基酸;HvBAS1 2长1 689 bp,编码562个氨基酸,二者氨基酸序列相似性为76.42%;亚细胞定位预测结果显示二者均存在于内质网。(3)实时定量PCR检测发现,青稞HvBAS1 1与HvBAS1 2的表达模式完全不同,其中HvBAS1 1在青稞根中的表达量高于叶中,而HvBAS1 2在叶中的表达量高于根,且随着苗龄的增大,HvBAS1 1在根中的表达呈先升高后降低的趋势,HvBAS1 2在叶片中呈先降低后升高的变化趋势;BRZ处理青稞幼苗后,其HvBAS1 1与HvBAS1 2基因的表达较对照均显著下调,且HvBAS1 2下调更为明显。研究表明,HvBAS1 1和HvBAS1 2很可能都参与了青稞内源BRs的失活,但二者的功能存在差异。     

Staphylococcus petrasii sp. nov. including S. petrasii subsp. petrasii subsp. nov. and S. petrasii subsp. croceilyticus subsp. nov., isolated from human clinical specimens and human ear infections

Thirteen coagulase-negative, oxidase-negative, and novobiocin-susceptible staphylococci were isolated from human clinical specimens. The isolates were differentiated from known staphylococcal species on the basis of 16S rRNA, hsp60, rpoB, dnaJ, tuf, and gap gene sequencing, automated ribotyping, (GTG)₅-PCR fingerprinting, and MALDI-TOF MS analysis. Phylogenetic analysis based on the 16S rRNA gene sequence indicated phylogenetic relatedness of the analyzed strains to Staphylococcus haemolyticus, Staphylococcus hominis, Staphylococcus devriesei, and Staphylococcus lugdunensis. DNA–DNA hybridization experiments between representative strains CCM 8418^T, CCM 8421^T, and the closest phylogenetic neighbors confirmed that the isolates represent novel Staphylococcus species, for which the name Staphylococcus petrasii sp. nov. is proposed. Genotypic and phenotypic analyses unambiguously split the strains into two closely related subclusters. Based on the results, two novel subspecies S. petrasii subsp. petrasii subsp. nov. and S. petrasii subsp. croceilyticus subsp. nov. are proposed, with type strains CCM 8418^T (=CCUG 62727^T) and CCM 8421^T (=CCUG 62728^T), respectively.     

Phytophthora captiosa sp. nov. and P. fallax sp. nov. causing crown dieback of Eucalyptus in New Zealand

A locally severe crown disease of exotic plantation Eucalyptus trees has been recorded periodically in New Zealand since 1986. Symptoms include leaf spots, petiole infection and twig and small branch lesions. Outbreaks of disease are episodic and individual trees may show marked variation in crown symptoms ranging from unaffected to total defoliation. Two previously unknown species of Phytophthora are associated with the disease. These are described and formally designated here as P. captiosa, from Eucalyptus botryoides and E. saligna; and P. fallax, from E. delegatensis, E. fastigata, E. nitens and E. regnans. Both P. captiosa and P. fallax have non-papillate, non-caducous sporangia and both are self-fertile. Phylogenetic analysis on the basis of ITS rDNA sequence data indicates they are closely related to each other but evolutionarily distant from the majority of described Phytophthora taxa. They share a common ancestor with another assemblage of Phytophthora lineages that includes P. insolita, P. macrochlamydospora and P. richardiae. Sporulation of P. captiosa and P. fallax has not been observed in the field. The mode of infection and spread of these non-caducous Phytophthora species in the eucalypt tree canopy remains unknown. This issue, and the possible geographic origins of these two Phytophthora species are discussed.     

Biodegradation of cyanide by Trichoderma spp. and Fusarium spp.

The rate of degradation of cyanide by certain strains of the Trichoderma spp. was evaluated. For comparison two Fusarium spp., which had previously been demonstrated to degrade metallocyanides were also studied. Studies were carried out to assess the rate of degradation using cyanide as the sole source of carbon or in the presence of glucose. Biodegradation was observed in flask cultures using cyanide as the sole carbon source. Strong evidence of cyanide biodegradation and co-metabolism emerged from studies with flask cultures where glucose was provided as a co-substrate. The rate of degradation of 2000 ppm CN⁻ was enhanced almost three times in the presence of glucose. A concomitant increase in fungal biomass was also observed in all the strains over the experimental period. Growth yield calculations performed provided values that were comparable to those reported in literature for one-carbon substrates.