不同种源桃金娘表型性状多样性研究

为解析桃金娘表型性状多样性及其种源间关系，该文以20个不同来源的桃金娘为研究对象，在同质园栽培条件下，对其营养器官和花器官表型性状进行观测，采用方差分析、变异分析、Shannon-Wiener多样性指数分析和聚类分析等方法，探讨不同种源桃金娘表型性状多样性。结果表明：(1)不同种源桃金娘表型性状存在显著差异(P<0.05),Shannon-Wiener多样性指数均值在1.35以上，表型性状多样性丰富。(2)种源内表型性状变异系数均值在10.81%～63.75%之间，种源间的变异系数均值在13.08%～74.04%之间，种源间变异(23.33%)高于种源内变异(19.79%),营养器官变异(29.52%)高于花器官变异(14.06%)。(3)部分性状存在极显著或显著相关性，株高与分枝数呈极显著负相关，而与叶长、叶宽和叶面积等却呈显著正相关。(4)在欧式距离10处，20个种源桃金娘可分为A、B、C三类，A类包含8个种源，该类种源表现为植株高大、分枝少、叶大和花大等特征；B类包含11个种源，该类种源表现为株高中等、叶较大和花中等等特征；C类仅包含1个种源，表现为植株低矮、分枝多、叶小和...     

桦褐孔菌提取物抗氧化活性研究

以乙酸乙酯和甲醇为提取剂,采用索氏提取法,剩余残渣采用热水浸提,最终得到桦褐孔菌不同极性提取物,对其DPPH自由基、羟基自由基及超氧阴离子自由基清除活性作用进行了研究,确定桦褐孔菌的抗氧化能力,为深入研究和开发桦褐孔菌功能性食品奠定理论基础。实验结果表明桦褐孔菌具有较好的抗氧化活性,其中乙酸乙酯提取物的DPPH自由基清除率、羟基自由基清除率和超氧阴离子自由基清除率均高于其他两组分及BHT,桦褐孔菌提取物有望成为功能性食品组分中合成抗氧化剂的天然替代品。     

不同种源花榈木苗期生长及生理特性比较

为了解不同种源花榈木在贵阳的生长特性和差异,该文通过对10个种源地花榈木进行育苗试验,测定其两年生实生苗的苗高、地径、生物量、叶片光合参数、光合色素、硝酸还原酶活性、硝态氮含量和根系活力,并进行差异性分析。结果表明:(1)10个种源花榈木净光合速率、气孔导度、胞间CO₂浓度、蒸腾速率和水分利用效率差异显著(P<0.05),表明不同种源花榈木光合特性及光能利用效率具有较大差异,浙江杭州和浙江永康花榈木是具有较高光合生长潜力的种源。(2)种源间的叶绿素含量、硝酸还原酶、硝态氮、根系活力存在显著差异,福建建瓯种源的叶绿素a、叶绿素b含量和叶绿素总量最高,能够将光合原初反应过程中积蓄的光能进行高效地传递,促进碳的同化; 贵州花溪种源硝酸还原酶活性最大,硝态氮含量最高,对氮元素的利用能力较强,能够促进植物蛋白质、氨基酸和叶绿素等的合成; 贵州望谟种源根系活力最大,吸收养分的能力强。(3)各种源间苗高、地径和生物量的分配存在显著差异,浙江杭州种源的植株枝叶繁茂、根系发达,生长表现好,安徽黄山种源的植株矮小,生长表现较差; 浙江杭州种源将生物量更多分配在根和叶,提高其根系吸收养分和叶片获取光能的能力,安徽黄山种源总体生物量积累最少,长势最差。(4)通过主成分分析法对各种源的花榈木适应性进行综合评价,结果显示浙江杭州种源>贵州黎平种源>浙江永康种源>贵州望谟种源>福建建瓯种源>贵州凯里种源>贵州石阡种源>贵州花溪种源>贵州平塘种源>安徽黄山种源。综上结果表明,浙江杭州、贵州黎平和浙江永康种源花榈木对贵阳地区立地环境具有较强的适应能力和生长潜力。     

桑黄液体发酵工艺的研究

通过对桑黄液体发酵培养基、培养条件优化实验研究,以获得具有与桑黄子实体相似功效成分的桑黄菌丝体液体发酵工艺。以菌丝体收率为主要考察指标,采用单因子及L9(34)正交实验的方法,对桑黄液体发酵培养基及培养条件进行优化,确定桑黄液体发酵工艺条件。桑黄液体发酵最佳培养基及培养条件:玉米粉2%,葡萄糖3%,酵母膏0.5%,蛋白胨0.5%,KH2PO40.3%,Mg SO4·7H2O 0.15%,VB120μg/100 m L,p H5.5,接种量8%,培养温度28℃,摇床转数180 r/min,培养周期82 h。优化条件下所获得桑黄菌丝体粉为土黄色,菌丝体平均得率为1.67%,菌丝体黄酮含量(0.84%)与桑黄子实体(0.88%)相当,菌丝体多糖含量(5.15%)是子实体(1.71%)的3倍。可见,该桑黄液体发酵工艺具有较大的推广应用价值。     

不同种源羊草表型差异性

研究植物因环境变化产生的表型差异,对于其适应未来全球气候变化具有重要的意义。羊草具有广泛的地理分布,是研究表型差异的理想材料。将白城市镇赉种源地羊草(ZL),白城市洮北种源地羊草(TB),吉林大安市种源地羊草(DA)和大安市姜家甸草场种源地羊草(JC)进行同质园移栽培养,分析了4个种源羊草的表型特征。结果表明,不同种源间羊草表型特征株高、叶长、叶宽、茎直径和生理特征蒸腾速率(Tr)、水分利用效率(WUE)和生物量分配均存在显著差异。在同质园环境培养下,这些表型特征均存在显著的种源差异。从总体上来看,TB种源地的羊草具有最高的株高、叶长和叶宽,分别为72.2 cm、34.4 cm、和10.34 mm。ZL种源有着最高的生物量(46.05 g)和WUE(3.06μmolCO₂/mmol H₂O)。ZL种源叶鲜重和干重均表现出了较高值,分别为2.88 g和0.96 g。不同种源羊草经移栽到同质园环境下,因为对源地气候长期的适应,产生了明显的表型差异,进而引起形态、生理特征的差异。这有利于羊草适应未来剧烈的气候变化,进一步揭示了羊草适应环境的策略。     

Molecular characterization of alanine racemase from Bifidobacterium bifidum

Bifidobacterium bifidum is a useful probiotic agent exhibiting health-promoting properties, and its peptidoglycans have the potential for applications in the fields of food science and medicine. We investigated the bifidobacterial alanine racemase, which is essential in the synthesis of -alanine as an essential component of the peptidoglycans. Alanine racemase was purified to homogeneity from a crude extract of B. bifidum NBRC 14252. It consisted of two identical subunits with a molecular mass of 50 kDa. The enzyme required pyridoxal 5′-phosphate (PLP) as a coenzyme. The activity was lost in the presence of a thiol-modifying agent. The enzyme almost exclusively catalyzed the alanine racemization; other amino acids tested, except for serine, were inactive as substrates. The kinetic parameters of the enzyme suggested that the B. bifidum alanine racemase possesses comparatively low affinities for both the coenzyme (9.1 μM for PLP) and substrates (44.3 mM for -alanine; 74.3 mM for -alanine). The alr gene encoding the alanine racemase was cloned and sequenced. The alr gene complemented the -alanine auxotrophy of Escherichia coli MB2795, and an abundant amount of the enzyme was produced in cells of the E. coli MB2795 clone. The enzymologic and kinetic properties of the purified recombinant enzyme were almost the same as those of the alanine racemase from B. bifidum NBRC 14252.     

Molecular characterization of Pasteurella multocida isolates from rabbits

Forty isolates of Pasteurella multocida from healthy (17 isolates) and diseased (23 isolates) rabbits were assayed for the presence of plasmids in seeking to determine whether any correlation exists between the presence of plasmids and health status, sensitivity to antimicrobial agents, capsular and somatic type, and the anatomic site of isolation. Six isolates were found harboring plasmids. A similar ladder pattern ranging from 18 to 3 megadalton (Mda) were found in three isolates recovered from diseased rabbits. One band of molecular weight 6.6 Mda was shared by four of five (4/5) isolates from the diseased rabbits. No correlation was found between the presence of the common plasmids and serotype, resistance to antimicrobial agents, and anatomic sites from which the bacteria were cultured. Random amplification polymorphic DNA was applied to subtype all the isolates of P. multocida. Two single primers were tested for their abilities to generate individual fingerprints by using PCR. Primer 1 grouped the isolates into 7 profiles, and primer 2 grouped them into 15. Random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) results show the presence of a wide heterogeneity within P. multocida isolates. Therefore RAPD-PCR is an efficient technique to detect the DNA polymorphism and could be used to discriminate P. multocida of rabbit isolates together with serologic typing.     

Novel hosts of the Eucalyptus canker pathogen Chrysoporthe cubensis and a new Chrysoporthe species from Colombia

The pathogen Chrysoporthe cubensis (formerly Cryphonectria cubensis) is best known for the important canker disease that it causes on Eucalyptus species. This fungus is also a pathogen of Syzygium aromaticum (clove), which is native to Indonesia, and like Eucalyptus, is a member of Myrtaceae. Furthermore, C. cubensis has been found on Miconia spp. native to South America and residing in Melastomataceae. Recent surveys have yielded C. cubensis isolates from new hosts, characterized in this study based on DNA sequences for the ITS and β-tubulin gene regions. These hosts include native Clidemia sericea and Rhynchanthera mexicana (Melastomataceae) in Mexico, and non-native Lagerstroemia indica (Pride of India, Lythraceae) in Cuba. Isolates from these hosts and areas group in the sub-clade of C. cubensis accommodating the South American collections of the fungus. This sub-clade also includes isolates recently collected from Eucalyptus in Cuba, which are used to epitypify C. cubensis. New host records from Southeast Asia include exotic Tibouchina urvilleana from Singapore and Thailand and native Melastoma malabathricum (Melastomataceae) in Sumatra, Indonesia. Consistent with their areas of occurrence isolates from the latter collections group in the Asian sub-clade of C. cubensis. DNA sequence comparisons of isolates from Tibouchina lepidota in Colombia revealed that they represent a new sub-clade within the greater Chrysoporthe clade. Isolates in this clade are described as Chrysoporthe inopina sp. nov., based on distinctive morphological differences.     

Genotype variation of Trypanosoma cruzi isolates from different Brazilian biomes

Chagas disease is an enzootic disease, in which the flagellate Trypanosoma cruzi infects a large variety of animals. Humans are accidentally infected due to the migration into wild environments. To identify T. cruzi discrete typing units (DTUs), 19 Brazilian isolates from different biomes and hosts were analyzed by PCR amplification of 24Sα rRNA, 18S rRNA and mini-exon gene sequences. The majority of the isolates was classified as TcIIb (TcII) but subtypes TcIIc (TcIII) and TcIId (TcV) were also identified. In addition, in monkeys TcI was detected.     

不同种源木姜叶柯种子生物学性状变异及质量特征研究

为探讨不同种源木姜叶柯种子生物学性状变异规律和质量特征,该文以8个省份10个种源为对象,测定了种子4个生物学性状、4个营养成分及3个质量性状数值,开展了11个种子特征参数的相关性及聚类分析。结果表明:(1)种子单粒重、种子长、种子数/种序、种序长均值依次为1.82 g、14.57 mm、16.43个、11.78 cm,不同种源种子 4个生物学性状变异均达到极显著水平。(2)种子蛋白质、脂肪、还原糖和淀粉均值分别为4.75 g/100 g、0.97 g/100 g、1.13%、74.23 g/100 g,淀粉在4个营养成分指标中差异最小。(3)种子百粒重、含水率和生活力均值依次为172.92 g、42.05%、53.00%,不同种源种子3个质量性状差异较大。(4)种子4个生物学性状与淀粉、蛋白质分别正向、负向相关,种子长与脂肪及还原性总糖正相关。(5)种子生物学性状与生境因子相关性复杂,种序长与无霜期极显著相关,相关系数达到0.832,与海拔显著负相关,相关系数达到-0.698,随纬度递增种子品质略变差。(6)种子聚类为第Ⅰ类短种序高营养成分型,第Ⅱ类长种序高生活力型,第Ⅲ类种子繁多高淀粉型等3类,聚类不呈现地理效应。综上所述,不同种源种子生物学性状和质量变异丰富,说明不同种源在子代分化已有遗传基础,种源筛选还需结合田间试验。     

Molecular characterization and antimicrobial resistance of Clostridium perfringens isolates of different origins from Costa Rica

Clostridium perfringens, a Gram positive, spore-forming anaerobe, is widely distributed in nature. Based upon their production of four major toxins alpha, beta, epsilon and iota, C. perfringens is classified into five toxinotypes (A-E). Some strains produce an enterotoxin (CPE), encoded by the cpe gene, which causes diarrhea in humans and some animals. C. perfringens strains that had been previously isolated and been kept at -80 degrees C were analyzed for the presence of toxin genes and for antimicrobial resistance: 20 from soils, 20 from animal, 20 from human origin and 21 from food non related to outbreaks. According to PCR results, all strains were classified as C. perfringens type A, since only alpha toxin gene was detected, while cpe was detected in two strains (2.5%) isolated from food, as it has been described in other world regions. Antibiotic resistance to at least one antibiotic was detected in 44% of the strains, 41% was resistant to clindamycin, 25% to chloramphenicol, 22% to penicillin and 20% to metronidazole. Soils strains showed the highest resistance percentages to almost all antibiotics. Multiresistance (to three or more antibiotic groups) was detected in the strains from soil (40%), human origin (30%), food (14%) and animal origin (5%). The high resistance rates found may be explained by the widespread use of antimicrobials as growth promoters in plants and animals; also these resistant strains may act as reservoir of resistance genes that may be transferred between bacteria in different environments.     

Variability in conidial thermotolerance of Metarhizium anisopliae isolates from different geographic origins

Notable variability in thermotolerance was found among conidia of 16 isolates of the insect-pathogenic fungi Metarhizium anisopliae var. anisopliae and one M. anisopliae var. acridum isolated from latitudes 61 degrees N to 54 degrees S. Conidial suspensions were exposed to 40 or 45 degrees C for 2, 4, 8, and 12 h. Most of the isolates tolerated 40 degrees C very well, with relative germination (germination relative to unheated controls) above 90% after 12 h of exposure. Exceptions were three isolates originating from high latitude, viz., ARSEF 2038 (38 degrees N, South Korea), 4295 (54.4 degrees S, Australia), and 5626 (61.2 degrees N, Finland) that had approximately 80% germination. High variability, however, was observed among isolates at 45 degrees C; viz., after 2 h exposure, relative germination was above 80% for six isolates, between 50 and 70% for three isolates, and between 0 and 30% for eight isolates. After 8 and 12 h at 45 degrees C, only two M. anisopliae isolates pathogenic to grasshoppers, viz., ARSEF 324 (latitude 19 degrees S, Australia) and 3609 (15 degrees N, Thailand), had high relative germination (91.6 and 79.4%, respectively, for 8 h exposures; and 90 and 47.1%, respectively, for 12 h). These isolates also were the most tolerant to UV-B radiation [J. Invertebr. Pathol. 78 (2001) 98-108]. The median lethal dose (LD50) for isolate ARSEF 324 was 49.4 and 47.9 degrees C, for 2 and 4 h of exposures, respectively. Exposure of conidia to wet-heat greatly delayed germination of some isolates. In general, isolates from higher latitudes demonstrated greater heat susceptibility than isolates from nearer the equator. Dry conidia tolerated 50 degrees C better than 45 degrees C in aqueous suspension.     

Molecular characterization of methanotrophic isolates from freshwater lake sediment

Profiles of dissolved O(2) and methane with increasing depth were generated for Lake Washington sediment, which suggested the zone of methane oxidation is limited to the top 0.8 cm of the sediment. Methane oxidation potentials were measured for 0.5-cm layers down to 1.5 cm and found to be relatively constant at 270 to 350 micromol/liter of sediment/h. Approximately 65% of the methane was oxidized to cell material or metabolites, a signature suggestive of type I methanotrophs. Eleven methanotroph strains were isolated from the lake sediment and analyzed. Five of these strains classed as type I, while six were classed as type II strains by 16S rRNA gene sequence analysis. Southern hybridization analysis with oligonucleotide probes detected, on average, one to two copies of pmoA and one to three copies of 16S rRNA genes. Only one restriction length polymorphism pattern was shown for pmoA genes in each isolate, and in cases where, sequencing was done, the pmoA copies were found to be almost identical. PCR primers were developed for mmoX which amplified 1.2-kb regions from all six strains that tested positive for cytoplasmic soluble methane mono-oxygenase (sMMO) activity. Phylogenetic analysis of the translated PCR products with published mmoX sequences showed that MmoX falls into two distinct clusters, one containing the orthologs from type I strains and another containing the orthologs from type II strains. The presence of sMMO-containing Methylomonas strains in a pristine freshwater lake environment suggests that these methanotrophs are more widespread than has been previously thought.     

Molecular characterization of Mycobacterium bovis isolates from Ethiopian cattle

Background

Bovine Tuberculosis (BTB) is a widespread and endemic disease of cattle in Ethiopia. Information relating to genotypic characteristics of Mycobacterium bovis strains affecting the cattle population in Ethiopia is limited. We carried out molecular characterization of M. bovis strains isolated from BTB infected cattle using the spoligotyping technique. The relationship between distribution of spoligotypes and recorded variables was also investigated. A new approach that can numerically reflect the degree of genetic polymorphism in a M. bovis population was also developed. The study was conducted from July 2006 to January 2007 in cattle slaughtered at five representative abattoirs in Ethiopia.

Results

Forty-five M. bovis isolates were obtained from 406 pathologic tissue specimens collected from 337 carcasses with lesions compatible with BTB. Twelve spoligotypes were identified from 34 distinct strains; with SB1176 as a dominant spoligotype (41.2% of the isolates) followed by SB0133 (14.7%). Comparison of spoligotypes with an M. bovis global database http://www.mbovis.org revealed six new spoligotypes which were subsequently registered in the database with international identification codes of SB1517, SB1518, SB1519, SB1520, SB1521 and SB1522. The majority of strains were obtained from cattle slaughtered at Addis Ababa abattoir. On the basis of the Spoligotype Evolutionary Index, SEI (a numeric expression approach to make standardized comparison of spoligotype evolution), M. bovis isolates from Ethiopia were relatively more heterogeneous (SEI = 3.2) compared to isolates from other countries. This might be attributed to extensive livestock movement linked to trading or seasonal migration, high degree of livestock mingling, and also diversities of the country's agricultural and livestock ecosystems, in addition to lack of disease control measures that led to high infection prevalence. Multiple spoligotype infection was recorded in nine (50%) of infected carcasses and this may indicate the prevailing high degree of super infection.

Conclusions

This study provided molecular evidence for the widespread distribution of M. bovis in the cattle population in Ethiopia. It also demonstrated a relatively high degree of genetic polymorphism of the isolates. Further molecular investigation of M. bovis strains in humans and other domestic animals is recommended in order to elucidate the zoonotic importance as well as reservoirs and pattern of transmission among various hosts.     

不同种源麻栎种子和苗木性状地理变异趋势面分析

麻栎(Quercus acutissima Carr.)是优良的能源和用材树种,在我国分布广泛并已有悠久栽培利用的历史,研究其种子和苗木性状地理变异规律对选择优良种源和适生区十分必要.研究收集了13个省(区)36个种源的麻栎种子,在测定麻栎种子性状的基础上,在安徽省滁州市红琊山林场进行了苗期试验和苗木性状测定,并采用趋势面分析方法对这些性状在经纬2维方向的地理变异模式进行了剖析.结果表明:1)麻栎种源间种子长度、宽度、百粒重和营养内含物存在极显著差异.种子百粒重、长度和宽度总体表现双向渐变趋势,随经度增高而增大,随纬度增高而减小,主要受到经度的控制;以西南到东北为中间地带,可溶性糖含量向东南表现先下降后上升的趋势,向西北则相反;淀粉含量从西北到东南呈逐渐减小的趋势.2)麻栎种源苗高、地径、生物量、热值和木材化学组分存在极显著差异.苗高、地径和生物量均呈双向渐变,经正向变异且变化幅度较大,纬负向变异且变化幅度较小,经度影响大于纬度;热值拟合回归方程不显著,方程无意义.木质素含量北部大于南部,但北部以西北部最高,南部以东南部最高.3)研究显示,麻栎种子和苗木性状多数存在显著的地理变异模式,这也是麻栎在长期进化过程中为适应复杂多变的环境而产生与之相适应的遗传变异结果.     

Variability in thermal responses among Furia gastropachae isolates from different geographic origins

The effect of temperature ranging from 5-30 degrees C on in vitro vegetative growth and conidial germination of isolates of the entomophthoralean fungus Furia gastropachae was investigated. Eleven isolates were used for growth studies; two from Maryland, six from New York, and three from Ontario. A subset of four isolates, one each from Maryland and New York and two from Ontario, were used in conidial germination experiments. Growth and germination were significantly associated with temperature for all isolates, occurring throughout the range 5-30 degrees C, though both processes were inhibited to varying degrees at upper and lower extremes. Temperature optima for growth ranged from 20 to 27 degrees C, and for germination from 20 to 25 degrees C. Although significant variability was observed among isolates in growth at temperatures above 13 degrees C, temperature optima were not significantly different among isolates, and variability did not appear to relate to the geoclimatic origins of the isolates. In contrast, germination responses to temperature did appear to be related to geographic origin. Furia gastropachae isolates from New York and Maryland germinated more slowly at 10 degrees C than did Ontario isolates, although the percentage of conidia ultimately germinating at each temperature was the same for all isolates. The New York and Maryland isolates performed much better at 30 degrees C, with significantly greater overall germination and secondary conidial discharge, than the Ontario isolates. Compared with other isolates at 30 degrees C, Ontario isolates were the least active, often failing to successfully discharge any secondary conidia.