Genetic variability of phosphate solubilizing activity by monocaryotic and dicaryotic mycelia of the ectomycorrhizal fungus Laccaria bicolor (Maire) P.D. Orton

Homocaryotic mycelia obtained from spores of Laccaria bicolor S238 have been compared in vitro for their efficiency in solubilizing poorly soluble phosphates. This could lead to genetic selection according to such criteria. However, there is very little room for improvement as the wild strain was shown to be one of the most efficient solubilizers among the strains tested. Twenty dicaryotic strains obtained by crossing the compatible homocaryons have also been compared and no clear heritability of this character has been found. The four phosphate salts used are most probably solubilized by the same mechanism which is polygenetically controlled     

Population structure and mycelial phenotypic variability of the ectomycorrhizal basidiomycete Laccaria bicolor (Maire) Orton

Mating type allele distribution and phenotypic variability were investigated in field populations of Laccaria bicolor. Sporophores associated with Norway spruce (Picea abies), colonized by natural sources of inoculum and growing in a seed orchard, were sampled to obtain dikaryotic strains and assay their phenotypic variability for traits important to the symbiosis. Basid-iospores were also collected for mating type analysis of different mycelia. Four sporophore mating types were identified containing seven A and five B factors. Out-breeding efficiency was estimated at 73.8% and the population was slightly inbred. Crosses with previously characterized L. bicolor strains from two nearby populations identified in total six sporophore mating types and ten A and nine B factors, for an estimated outbreeding efficiency (85.7%) similar to previous studies of more spatially disparate Laccaria spp. populations. Dikaryotic strains were tested for mycelial growth rate, as a measure of their competitive ability, on agar media containing a soluble (NaH₂PO₄), or an insoluble (CaHPO₄) phosphate source. Their ability to solubilize the latter was also tested to assess their relative capacity to access insoluble, inorganic phosphate. In most cases, significant variation was detected among strains from the same site for all variables. On three sites (VC4, VC5 and VC7), each determined previously to possess a uniform mycelial genotype, phenotypic variability was likely due to epigenetic variation among different strains of the same genotype. Possible evidence for dikaryon-monokaryon crosses was observed in vivo on one sample site (VC2) where adjacent mycelia shared two mating factors. The phenotypic variability of different mycelial genotypes reflected their genetic variability observed as mating type allele diversity and underlined the importance of basidiospore dispersal in introducing new genotypes into the population. The reproductive strategies of L. bicolor are discussed and compared to those of other basidiomycete species.     

荧光蛋白在双色蜡蘑中的构建与表达

菌根是真菌与植物之间形成的互惠互利的营养共生体,对生态环境有重大的意义。外生菌根真菌与植物互作机制以及真菌基因功能的深入研究都需要对菌根真菌进行遗传转化,本研究以外生菌根真菌模式生物双色蜡蘑(Laccaria bicolor)为研究对象,选择细胞核中的核小体蛋白H₂B基因为目的基因,以pCEBN为表达载体,融合红色荧光蛋白,最终构建在真菌中表达的双元载体,使用根瘤农杆菌介导转化法转化双色蜡蘑菌丝,随后利用PCR对真菌转化子进行验证后,通过激光共聚焦显微镜观察到菌丝细胞核中的红色荧光,成功将融合荧光蛋白转化菌根真菌,为后续研究菌根真菌中基因的亚细胞定位提供了实验平台。结果表明,利用双元载体和农杆菌转化方法,建立了高效的双色蜡蘑转化体系(93.33%),在激光共聚焦显微镜下观察到菌丝细胞核中红色荧光信号,验证了融合荧光蛋白在双色蜡蘑中的成功表达。本研究成功地构建了菌根真菌中的核小体蛋白和红色荧光蛋白融合表达的真菌转化体系。     

双色蜡蘑对黑松幼苗生长量及其根系形态的影响

为研究双色蜡蘑(Laccaria bicolor)对黑松(Pinus thunbergii)幼苗生长及其根系形态的影响,在营养杯培育条件下,用双色蜡蘑液体菌剂对黑松幼苗进行接种处理,接种第15、30、60、90、120天时取样,比较接种和未接种黑松幼苗的生物量、根系形态及根系分形维数的差异。结果表明:双色蜡蘑在黑松幼苗地上植株、地下根系的生长及其生物量的积累方面都表现出明显的促进作用。接种双色蜡蘑也显著改善了根系总长度、分支数、表面积、体积等参数和根系分形维数,并对地下根系生长的促进作用时间早于地上部分,且效应显著高于地上部分。接种双色蜡蘑第15~30天时对地上部分基本无影响,但对根系促进作用明显,而地上部分在第60天时开始表现出显著的生长效应。研究发现,双色蜡蘑能够成功定殖于黑松根部,促进黑松幼苗生长及其生物量的积累,同时显著促进根系总长度、分支数、表面积和体积增加,并使根系分形维数增大,表现出明显的促生作用,且对根系发育的显著促进作用早于地上部分。     

Dibenzocyclooctane lignans from the stems of Schisandra bicolor

A new dibenzocyclooctane lignan, schisanbicolorin A, together with fifteen known lignans, were isolated from the stems of Schisandra bicolor Cheng. Their structures were identified as (aS,6R,7S)-5,6,7,8-tetrahydro-2,3,13-trimethoxy-6,7-dimethyldibenzo[3,4]cycloocta [1,2-f][1,3]benzodioxol-1-ol (1), neglschisandrin C (2), angeloylgomisin R (3), schisantherin D (4), gomisin F (5), schisantherin B (6), tigloylgomisin Q (7), gomisin G (8), interiotherin B (9), schisandrin (10), angeloylgomisin H (11), benzoylgomisin H (12), gomisin H (13), angeloyl−(+)−gomisin K₃ (14), deoxyschizandrin (15), and (+)-gomisin K₃ (16), respectively, based on spectroscopic analysis and by comparison of their spectral data with those reported previously in the literature.     

油松根际外生菌根真菌龟裂秃马勃的分离鉴定及培养

该研究采用组织分离法对黄土高原子午岭林区连家砭林场油松根际外生菌根真菌进行了分离,并采用ITS序列对分离的真菌进行鉴定,研究了菌根真菌的培养条件,为菌剂的生产奠定基础。结果表明:子午岭林区油松根际分离培养出的真菌为龟裂秃马勃(Handkea utriformis),能够与油松形成外生菌根。采用MMN培养基为基础培养基,纯培养条件下,龟裂秃马勃生长的最适pH为6.0,最适温度为25℃。以蔗糖为唯一碳源的培养基中龟裂秃马勃生长速度最快,其次是麦芽糖和葡萄糖,生长最慢的为乳糖,在以几丁质和碳酸氢钠为碳源的培养基上不生长。有机氮源中,对脯氨酸为唯一氮源的利用能力最好,其次是酪蛋白,尿素最差;无机氮源中,以磷酸氢二铵为唯一氮源时生长最好,其次是硝酸钙,最差为酒石酸铵。该研究首次报道了黄土高原子午岭林区外生菌根真菌龟裂秃马勃,并初步确定了其最佳培养条件。     

我国首次检出双色多刺蚁及多刺蚁属介绍

【目的】2016年7月北京出入境检验检疫局从泰国进境商品山竹果中检出双色多刺蚁,经查询,该种为我国进出境检疫部门首次检出。了解多刺蚁属情况可为该类昆虫的检疫鉴定提供依据。【方法】笔者通过对双色多刺蚁及多刺蚁属相关文献进行翻译和整理,详细介绍了双色多刺蚁的分类地位、分布、形态特征及近似种,以及多刺蚁属在中国的种类,并简要分析了我国检疫系统对多刺蚁属的截获情况。【结果】双色多刺蚁(膜翅目:蚁科)主要分布于东南亚和澳大利亚地区,在我国仅分布于云南省。多刺蚁属在我国已知45种1亚种。2003—2016年7月我国检疫系统共检出多刺蚁属2097批次。【结论】多刺蚁属随货物包装携带入境的概率较高,各进出境检疫部门应加强对该类昆虫的检疫工作。     

高粱磷脂酶D基因家族的鉴定和表达分析

磷脂酶D（PLD）是植物生长和胁迫反应过程中参与膜磷脂分解代谢的关键酶。PLD编码基因在高等植物中构成了一个大的基因家族，但是仍未对高粱PLD基因家族进行深入研究。本研究中，通过全基因组分析，鉴定了15个PLD基因，并分成6个亚组，初步揭示了SbPLDs的基因结构、保守结构域、染色体定位和系统发育关系等信息。基因复制的研究表明，片段复制在高粱PLD基因家族的扩展中发挥了重要作用，SbPLDs在进化过程中经历了强烈的纯化选择。此外，转录组数据表明，受启动子区上游顺式元件调控的PLD家族基因可能在高粱的生长发育中发挥重要作用。通过real-time PCR分析，显示部分SbPLDs参与非生物胁迫和激素途径的潜力。亚细胞定位表明，高粱PLD蛋白在细胞质中富集，可能具有抗逆胁迫的功能。本研究为了解高粱PLD基因的特征提供了理论参考，为研究高粱PLD基因家族的进化提供了新的视角，也为阐明高粱PLD基因家族的功能提供了基础信息。     

紫背天葵叶片中花青素种类及合成调控基因转录组分析

为获取紫背天葵(Cynura bicolor DC.)叶片中花青素种类及其合成调控基因等信息,该试验以紫背天葵叶背面紫色以及经处理叶背面几乎全绿(对照)的叶片为材料,进行转录组测序分析,同时进行6个相关差异表达基因的qRT-PCR分析和6种花青素苷元的HPLC检测,以揭示紫背天葵特有的花青素苷元及其合成调控关键基因信息。结果表明:(1)在紫背天葵中共获得14个花青素苷元及32个花青素合成调控基因信息,其中表达量差异显著下调的4个基因为Pg(c11692)、Cy(c42112)、ANS(c38551)和3GT(c9064),表达量差异显著上调的2个基因是D FR(c35961)和3GT(c20283)。(2)qRT-PCR检测结果显示,上述6个基因在2种紫背天葵叶中的表达趋势(上调或下调)与转录组测序结果完全一致,但转录组测序检测到的表达趋势差异倍数比qRT-PCR检测结果更加明显。(3)HPLC分析显示,紫背天葵叶中均未检测到Dp、Pt、Pn及Mv等4类花青素苷元,但紫背天葵叶中富含Cy花青素苷元,且背面紫色的叶中Cy类花青素苷元含量(62.21 mg/kg)显著高于绿色叶对照(6.86 mg/kg);背面紫色和全绿叶中的Pg花青素苷元含量均低于0.43 mg/kg。研究推测,Cy和Pg花青素苷元在绿叶紫背天葵(对照)中含量显著降低可能是因为存在1个ANS和1个3GT正调控以及1个DFR和1个3GT负调控所致。     

Effect of salinity and abscisic acid on accumulation of glycinebetaine and betaine aldehyde dehydrogenase mRNA in Sorghum leaves (Sorghum bicolor)

The effects of salt stress and abscisic acid (ABA) on the expression of betaine aldehyde dehydrogenase (BADH) were determined in sorghum (Sorghum bicolor L.) plants. BADH mRNA expression was induced by salinity, and the timing coincided with the observed glycinebetaine (betaine) accumulation. The leaf water potential in the leaves of the sorghum plants was significantly affected by salinity. In response to salinity, betaine, ABA, Na and Cl accumulations increased 6-, 16-, 90-, and 3-fold, respectively. In the leaf disks from unsalinized plants incubated on NaCl, or ABA solution, the BADH mRNA level was lower than in the ABA-treated disks. Exogenous application of the ABA biosynthetic inhibitor fluridone to the NaCl-treated disks reduced the ABA accumulation and BADH mRNA levels compared with NaCl-treated leaves. The results indicate that the salt-induced accumulation of betaine and BADH mRNA coincides with the presence of ABA.     

紫背天葵叶和花中花青素合成相关转录组基因分析

为获取紫背天葵(Gynura bicolor D C.)花青素合成代谢相关调控基因信息,该试验以紫背天葵叶片为材料,以其花朵为对照,进行转录组测序,并进行CHS、CHI、F3H等8类合成酶基因以及MYB、bHLH及WD40等3类转录因子检索,从中选取8个相关差异表达显著调控基因进行qRT-PCR验证分析。结果显示:(1)在紫背天葵中共获得72个花青素合成酶信息,其中差异表达明显的有1个F3′H和2个3GT下调,9个F3H基因中有上调基因4个和下调基因5个。(2)在紫背天葵中获取到238个MYB、113个bHLH和219个WD40转录因子,这3类转录因子中差异表达明显的分别为22个、16个和7个。(3)qRT-PCR结果显示,所选取的8个花青素合成相关调控基因,在紫背天葵叶及花朵中的下调表达趋势与转录组测序结果完全一致,但不同基因差异表达趋势略有不同。研究表明,在紫背天葵叶片和花朵中所存在的大量花青素合成代谢调控基因中,只有少量差异表达显著,但转录因子相比合成酶的调控更为复杂。     

Identification of alkaline stress-responsive genes of CBL family in sweet sorghum (Sorghum bicolor L.)

Calcineurin B-like proteins play important roles in the calcium perception and signal transduction of abiotic stress. In this study, the bioinformatic analysis of molecular characteristics of Sorghum bicolor calcineurin B-like protein (SbCBL) revealed that sequences of SbCBL are highly conserved, and most SbCBLs have three typical EF-hands structures. Among the SbCBL proteins, four of which, SbCBL01, 04, 05, 08, have a conserved N-myristoylation domain. Stress-responsive and phytohormone-responsive cis-elements were found in the promoter regions of SbCBL genes. Real-time quantitative polymerase chain reaction (RTqPCR) analysis showed that SbCBL genes have different tissue-specific expression patterns under normal growth conditions in sweet sorghum (Sorghum bicolor L. Moench). Interestingly, when treated with sodium carbonate, SbCBL genes also show various sodium carbonate stress responsive patterns in sweet sorghum seedlings. These results suggest that SbCBLs may participate in regulating sodium carbonate stress-specific cellular adaptation responses and influencing growth and developmental patterns in sweet sorghum.     

Effect of the polycyclic aromatic hydrocarbon phenanthrene on root exudation of Sorghum bicolor (L.) Moench

The objective of this work was to study the effect of two concentrations (10 and 100 mg kg⁻¹) of phenanthrene, a ubiquitous polycyclic aromatic hydrocarbon (PAH), on root exudation of the remediating plant Sorghum bicolor (L.) Moench under controlled conditions in a pot experiment. It was found that the phenanthrene concentration of 10 mg kg⁻¹ did not cause significant effects on plant survival and growth but had little stimulating effect on carbohydrate exudation. The contamination with phenanthrene at 100 mg kg⁻¹ inhibited accumulation of plant shoot and root biomass, decreasing the carboxylic acid, carbohydrate, and amino acid amounts released by sorghum root into the rhizosphere. However, root exudation per unit of root surface was not changed significantly with increasing phenanthrene concentration. There were no differences in qualitative composition of root exudates under the influence of PAH were found. The observed alterations in the ratio between the main root-exuded components are assumed to manifest adaptive alterations occurring in the plant as a response to pollutant stress. The activity of three oxidoreductases (oxidase, peroxidase, and tyrosinase) released by sorghum roots was clearly progressive to the increasing phenanthrene concentration in the substrate. Under the influence of phenanthrene, the population of phenanthrene-degrading microorganisms in sorghum root zone increased, and their share in the total number of culturable heterotrophs increased as well. The main promotional factor was the pollutant; however, the stimulating effect of the plant root exudates was also involved. The increased pollutant-degrading microbial population and activity of the extracellular root enzymes are presumed to be important for the rhizodegradation of PAH.     

中国万代兰属(兰科)一新记录种——双色万代兰

该文报道了在中国西藏林芝地区墨脱县发现的中国兰科(Orchidaceae)万代兰属(Vanda W.Jones ex R.Br.)一新记录种——双色万代兰(Vanda bicolor Griff.)。双色万代兰与白柱万代兰(V.brunnea Rchb.f.)的形态较近似,但前者唇瓣中裂片正面粉色或黄绿色,不具纹,先端稍开裂,易与后者相区别。同时还描述了该新记录种的形态特征,并提供了彩色图版。该研究为兰科植物的研究提供了基础,对丰富我国兰科植物物种多样性具有一定意义。     

Transmission of acquired adjustments to salinity in Sorghum bicolor

Previous studies showed that exposure of eight-day-old Sorghum bicolor for three weeks to sublethal salinity induces an increase in salinity tolerance, called physiological adaptation (A). During A, plants of a same population differ in reaction and tolerance to salinity. Tolerance levels of the reaction types depend on environmental conditions besides salinity. Reactions observed most frequently in an experiment have generally highest tolerance levels. This phenomenon is defined adaptive determinism (AD). In this study, the relationship between a potential source of the information subjacent to AD and AD itself is analysed in plants first exposed to salt-inducing A. When the reaction types are close variations of one reaction mode, AD is highest. This relationship is inversed in progeny of adapted plants. Results suggest that information relevant to AD is transmitted to the progeny of adapted plants, and that adaptive information is created during A in plants first exposed to adaptation inducing treatment.     

Purification and Properties of the Threespine Stickleback (Gasterosteus aculeatus) Lactate Dehydrogenase LDH-B4 and LDH-C4 Isoenzymes

In the threespine stickleback (Gasterosteus aculeatus) lactate dehydrogenase (LDH, EC 1.1.1.27) is encoded by three loci, Ldh-A, Ldh-B, and Ldh-C. LDH-B₄ isoenzyme restricted its function to eye and brain, while LDH-C₄ isoenzyme functions in the eye. In the Dead Vistula stickleback population, none of LDH loci is polymorphic. The LDH-B₄ and LDH-C₄ isoenzymes from the eye were purified to homogeneity to specific activity of 186 and 229 μmol NADH min⁻¹mg⁻¹, respectively, at 30°C. Some physico-chemical and kinetic properties revealed that eye LDH-C₄ isoenzyme was more thermostable and had a higher affinity to pyruvate than LDH-B₄ isoenzyme. Lower Km for pyruvate of eye LDH-C₄ isoenzyme distinguishes it from fish LDH-C₄ isoenzyme isolated from liver.