Evaluation of the Entomopathogenic Fungi Beauveria bassiana and Paecilomyces fumosoroseus for Microbial Control of the Silverleaf Whitefly, Bemisia argentifolii

Collaborative research was conducted at the USDA-ARS Subtropical Agricultural Research Center in southern Texas to assess the microbial control potential of Beauveria bassiana and Paecilomyces fumosoroseus against Bemisia whiteflies. Laboratory assays demonstrated the capacity of both pathogens to infect Bemisia argentifolii nymphs on excised hibiscus leaves incubated at relative humidities as low as 25% at 23 ± 2°C (ca. 35% infection by B. bassiana and P. fumosoroseus resulted from applications of 0.6–1.4 × 10³ conidia/mm² of leaf surface). In small-scale field trials using portable air-assist sprayers, applications at a high rate of 5 × 10¹³ conidia in 180 liters water/ha produced conidial densities of ca. 1–2.5 × 10³ conidia/mm² on the lower surfaces of cucurbit leaves. Multiple applications of one isolate of P. fumosoroseus and four isolates of B. bassiana made at this rate at 4- to 5-day intervals provided >90% control of large (third- and fourth-instar) nymphs on cucumbers and cantaloupe melons. The same rate applied at 7-day intervals also provided >90% control in zucchini squash, and a one-fourth rate (1.25 × 10¹³ conidia/ha) applied at 4- to 5-day intervals reduced numbers of large nymphs by >85% in cantaloupe melons. In contrast to the high efficacy of the fungal applications against nymphs, effects against adult whiteflies were minimal. The results indicated that both B. bassiana and P. fumosoroseus have strong potential for microbial control of nymphal whiteflies infesting cucurbit crops.     

Effects of selected pesticides used against glasshouse tomato pests on colony growth and conidial germination of Paecilomyces fumosoroseus

Two isolates of a potential biological control agent, Paecilomyces fumosoroseus, for greenhouse whitefly, Trialeurodes vaporariorum, on tomato in greenhouses were tested for their compatibility with five fungicides (captan, dichlofluanid, iprodione, benomyl, and carbendazim) and six insecticides (malathion, carbaryl, primicarb, pymetrozine, cypermethrin, and deltamethrine) which are used on glasshouse tomato. Conidial germinations after 24 h and radial growths after 14 days of inoculation were determined in the presence of each pesticide at three concentrations (recommended rate, 0.1 and 10 times of the recommended rate by its manufacturer) and resultant variations were evaluated. Our results suggest that the tested pesticides, especially the fungicides, have potential to interfere with the success of P. fumosoroseus if they are to be employed together. The recommended concentration rate of primicarb and pymetrozine, and the lower concentrations of cypermethrin and deltamethrin were less hazardous than the others.     

高雄山虫草及其细脚拟青霉无性型

报道采自安徽省天堂寨自然保护区的高雄山虫草Cordyceps takaomontana及其无性型细脚拟青霉Paecilomyces tenuipes ,应用单子囊孢子分离鉴定和微循环产孢的方法确证了两者的对应关系,并修订了高雄山虫草的原始描述。双梭孢虫草C．bifusispora可能为高雄山虫草的同物异名。同时采集的具数个红色子座且有双梭形子囊孢子的虫草不是同一个种。     

高雄山虫草及其细脚拟青霉无性型

报道采自安徽省天堂寨自然保护区的高雄山虫草Cordyceps takaomontana及其无性型细脚拟青霉Paecilomyces tenuipes,应用单子囊孢子分离鉴定和微循环产孢的方法确证了两者的对应关系,并修订了高雄山虫草的原始描述。双梭孢虫草C．Bifusispora可能为高雄山虫草的同物异名。同时采集的具数个红色子座且有双梭形子囊孢子的虫草不是同一个种。     

淡紫拟青霉固体发酵工艺研究

淡紫拟青霉（Paecilomyces lilacinus）是控制植物病原线虫的优良生防菌,同时对多种土传尖孢镰刀菌引起的枯萎病等病害具有防病作用。固态发酵技术为分生孢子的大规模、低成本生产提供了许多有利条件。为获得淡紫拟青霉固体发酵的最佳条件,采用单因素实验和响应面试验对淡紫拟青霉固体发酵培养基的组成、料液比、种子液接种量和烘干条件进行了优化,结果表明淡紫拟青霉固体发酵最佳的培养基组成为麸皮玉米粉为 0.9：1（体积比）、蔗糖添加量4%（质量分数,下同）、尿素添加量 0.16%、硫酸铵添加量 0.17%、料水比为1：0.5（体积比）,培养条件为接种量10%、培养温度28 ℃、培养时间6 d、固体菌剂最适烘干条件为35 ℃烘干24 h,在此条件下淡紫拟青霉固体菌剂的有效活菌数为 2.49×10¹⁰ cfu/g。     

蝉拟青霉液体发酵条件优化

采用液体发酵蝉拟青霉,对蝉拟青霉的发酵条件进行优化,以提高蝉拟青霉胞外多糖产量及生物量。摇瓶发酵条件下,在单因素基础上设计正交实验确定各因素的最佳组合。优化后得最佳发酵培养基:蔗糖8%,牛肉膏0.75%,酵母膏0.125%,MgSO_4·7H_2O 0.3%,KH_2PO_4 0.2%,麸皮0.5%。该条件下胞外多糖产量为5.96 g/L,生物量为42 g/L,较优化前提高了1倍。采用发酵罐进行扩大培养,对分批发酵时的初糖浓度进行了优化,并分析了补料分批发酵对发酵过程的影响。发酵罐培养时最适初糖浓度为5%,此时生物量最高为38 g/L,多糖含量最高为5.5 g/L;采用补料分批发酵时,多糖产量最高为5.89 g/L,生物量最高为40 g/L,效果优于分批发酵。     

Intraspecific genotypic diversity in plants

Variations in the nuclear DNA, mainly as a result of quantitative modulations of DNA repeats belonging to different sequence families of satellite DNA and to the activity of transposable elements, have been assessed within several angiosperm species. These variations alter the amount and organization of the DNA and therefore the genotype, rather than the genome proper. They take place on an evolutionary time scale as the result of selection processes after the occurrence of uncontrolled events in the genome or may be due to direct responses of plant genomes to environmental stimuli that occur under plant-level control within a short developmental period of a single generation. These DNA changes are correlated to changes in the developmental dynamics and phenotypic characteristics of the plants, and the capability to carry out genotypic variation is an evolutionary trait that allows plant species to adapt to different environmental conditions, as well as to the variability of conditions in a given environment. The link between developmental and environmental stimuli and repetitive DNA that elicits the intraspecific diversity of plant genotypes may provide models of evolutionary change that extend beyond the conventional view of evolution by allelic substitution and take into account epigenetic effects of the genome structure.     

等离子体法诱变淡紫拟青霉产几丁质酶菌种的筛选

采用菌株诱变技术,提高生防用淡紫拟青霉菌株产几丁质酶的能力。通过常温常压等离子体诱变技术(MPMS)对淡紫拟青霉进行诱变育种处理,对处理的菌种先采用透明圈法进行初筛,然后采用发酵方法进行复筛。采用MPMS法诱变淡紫拟青霉产几丁质酶菌种时,温度25℃,处理时间30 s,样品处理量60μL,诱变菌的致死率为30.33%时,正突变率为14%。采用摇瓶分批发酵培养,诱变菌种的几丁质酶活为0.17 U/mL。结果表明,经过对淡紫拟青霉的诱变处理,获得高活性几丁质酶产生菌株,几丁质酶酶活提高180%。     

大别山五针松种内和种间竞争强度

通过对安徽省岳西县大别山五针松群落内的53株对象木及2079株竞争木的调查,运用Hegyi单木竞争模型分析了大别山五针松的种内和种间竞争强度。结果表明,大别山五针松的种内和种间竞争强度分别为17.11%和82.89%,说明竞争主要来自种间。大别山五针松的伴生种较多,种内与主要伴生种间的竞争关系为短柄枹大别山五针松种内黄山松满山红茅栗金缕梅紫茎四照花灯台树鹅耳枥。竞争强度随对象木胸径的增大而减小,当对象木的胸径小于25cm时,所受到的竞争强度较大;当胸径在25cm以上时,竞争强度变化很小,二者符合幂函数关系(CI=AD-B),所得的预测模型能很好的预测大别山五针松种内和种间竞争强度。     

Effects of Paecilomyces lilacinus protease and chitinase on the eggshell structures and hatching of Meloidogyne javanica juveniles

A serine protease and an enzyme preparation consisting of six chitinases, previously semi-purified from a liquid culture of Paecilomyces lilacinus strain 251, were applied to Meloidogyne javanica eggs to study the effect of the enzymes on eggshell structures. Transmission electron microscopic studies revealed that the protease and chitinases drastically altered the eggshell structures when applied individually or in combination. In the protease-treated eggs, the lipid layer disappeared and the chitin layer was thinner than in the control. The eggs treated with chitinases displayed large vacuoles in the chitin layer, and the vitelline layer was split and had lost its integrity. The major changes in the eggshell structures occurred by the combined effect of P. lilacinus protease and chitinases. The lipid layer was destroyed; the chitin layer hydrolyzed and the vitelline layer had lost integrity. The effect of P. lilacinus protease and chitinase enzymes on the hatching of M. javanica juveniles was also compared with a commercially available bacterial chitinase. The P. lilacinus protease and chitinase enzymes, either individually or in combination, reduced hatching of M. javanica juveniles whereas a commercial bacterial chitinase had an enhancing effect. Some juveniles hatched when the eggs were exposed to a fungal protease and chitinase mixture. We also established that P. lilacinus chitinases retained their activity in the presence of endogenous protease activity.     

Intraspecific variation ofDrosophila buzzatii larval breeding success onOpuntia: A yeast-plant-insect relationship

The cactophilic species,Drosophila buzzatii, normally breeds in decaying pockets ofOpuntia cladodes, in which there is a complex interaction with the microbial flora, especially yeast species. Isofemale lines were used to estimate genetic variation among larvae reared on their natural feeding substrate. Four naturally occurring cactophilic yeast species isolated from the same Tunisian oasis as theDrosophila population were used. Two fitness components were studied for each line, viability and developmental time. Genetic variations amongD. buzzatii lines were observed for both traits. A significant yeast species x isofemale line interaction for viability was also evidenced, suggesting the occurrence of specialized genotypes for the utilization of breeding substrates. This genetic heterogeneity in the natural population may favor a better adaptation to the patchily distribution of yeasts.     

Degradation and enzymatic activities of three Paecilomyces inflatus strains grown on diverse lignocellulosic substrates

This paper describes for the first time in detail the lignocellulose degradation system in Paecilomyces inflatus. The fungal genus Paecilomyces contributes the carbon turnover from lignin and carbohydrate plant residues, particularly in compost and soil environment, where basidiomycetes appear very seldom. We studied three different strains of P. inflatus, obtained from different ecophysiological and geographical origin. Various cultivation conditions were employed, and the chemical analysis of decayed straw, compost, birch and spruce wood chips indicated variable responses. Endoglucanase, xylanase and laccase were assayed. All strains of P. inflatus, regardless of their origin, altered the ambient pH in a similar manner in all investigated substrates, suggesting that all P. inflatus isolates may share the common regulatory system to control their environmental pH. The variability among strains of P. inflatus in their ability to remove lignocellulose components often was related to the nature of the substrate and the production of specific enzymes although it was not strictly correlated. This may implicate that other enzymes and/or even other parameters needed for lignocellulosics degradation in P. inflatus should be evaluated. Indications for specific adaptation strategies that may operate in P. inflatus were found.     

球孢白僵菌种内比较线粒体基因组学分析

【目的】明确球孢白僵菌种内线粒体基因组的分化程度。【方法】从GenBank下载已知的球孢白僵菌6个菌株线粒体基因组序列,详细分析基因组的组成结构,比较外显子区、内含子区和基因间区的碱基变异情况,分析菌株间的系统发育关系。【结果】球孢白僵菌不同菌株的线粒体基因组大小为28.8–32.3 kb,都有14个常见的核心蛋白编码基因、2个rRNA基因和25个tRNA基因,具有很强的共线性关系。但是,不同菌株含有的线粒体内含子数目存在差异(2–5个/菌株),在cox1、cox2和nad1基因中表现出内含子插入/缺失多态性,这是导致线粒体基因组大小变化的主要因素。对外显子、内含子和基因间区的碱基变异情况进行分析,发现内含子和基因间区相对变异较大,而外显子区相对变异较小。系统发育分析发现,这些球孢白僵菌菌株以很高的支持度聚在一起,具有相同内含子分布规律的菌株也具有较近的聚类关系。【结论】本研究首次报道球孢白僵菌因内含子数目不同、插入缺失突变和单核苷酸变异等在线粒体基因组上表现出较大程度的遗传分化,为认识真菌种内线粒体基因组分化提供了新的证据。     

Intraspecific diversity and dominant genotypes resist plant invasions

Numerous studies have asked whether communities with many species deter invasions more so than do species-poor communities or whether dominant species deter invasion by colonizing species. However, little is known about whether high intraspecific diversity can deter biological invasions or whether particular genotypes might deter invasions. In this study, we present experimental evidence that intraspecific diversity and particular genotypes of tall goldenrod, Solidago altissima , can act as a barrier to colonization by new species. We found that biomass of colonizing species was negatively correlated with genotypic diversity, and particular genotypes affected the richness, cover, and biomass of colonizing species. Stem density of S. altissima increased with genotypic diversity and varied among genotypes, suggesting that stem density is a key mechanism in limiting colonization dynamics in this system. Our results indicate that the loss of intraspecific diversity within a dominant plant species can increase susceptibility to plant invasions.     

Intraspecific differences in the pattern of hibernation in the ground squirrelSpermophilus beldingi

Summary Intraspecific differences in the patterns of heterothermy were found in captive Belding's ground squirrels that hibernated undisturbed at ambient temperatures of 5°C, 10°C, and 15°C. The timing of all entrances into and arousals from hibernation was determined from records of copper-constantan thermocouples that were mounted on the floor of each animal's nest box and connected to continuously recording potentiometers. In the absence of food, large adult males terminated hibernation spontaneously in the spring. In contrast, females and small non-breeding males (yearlings) did not stop hibernating but instead they shortened their bouts of torpor in the spring so that they aroused every three or four days. This interval of frequent arousals, termed the emergence period, lastel until the squirrels became emaciated, and it was only in the 2 or 3 weeks preceeding death (starvation period) that arousal frequency once again decreased towards midwinter values (Fig. 3). These animals terminated hibernation when fed during the emergence or starvation periods, but they were able to resume torpor if that feeding lasted less than a week.Termination of hibernation and reproductive development in males were related to the size of the animals, not their age. Males two years and older which did not deposit normal quantities of fat were like yearlings in that they had open-ended hibernation seasons and showed little testicular enlargement. Likewise, juvenile males that grew to near adult size in their first summer spontaneously terminated hibernation and had well developed testes like most older individuals. In addition, both the frequency and duration of arousals in the emergence period were related, in part, to the extent of the animals' fat reserves, such that large squirrels spent more time at high body temperatures than small individuals (Figs. 5, 6). This trend was most pronounced at high ambient temperatures.These intraspecific differences in hibernation physiology are consistent with the sex and agerelated differences in the timing of emergence above ground in nature. The increase in the time spent euthermic in the spring isinterpreted as an adaptation for increasing the opportunities for environmental assessment. The intraspecific differences in the extent of this euthermia appear to be associated with differences in the balance between the energy supplies available to an animal and its need to accelerate the use of that energy (i.e., arouse) in order to achieve an early and accurately-time emergence from the hibernaculum.     

Intraspecific Phylogeography of Lacerta vivipara and the Evolution of Viviparity

The lacertid lizard Lacerta vivipara is one of the few squamate species with two reproductive modes. We present the intraspecific phylogeny obtained from neighbor-joining and maximum-parsimony analyses of the mtDNA cytochrome b sequences for 15 individuals from Slovenian oviparous populations, 34 individuals from western oviparous populations of southern France and northern Spain, 92 specimens from European and Russian viviparous populations, and 3 specimens of the viviparous subspecies L. v. pannonica. The phylogeny indicates that the evolutionary transition from oviparity to viviparity probably occurred once in L. vivipara. The western oviparous group from Spain and southern France is phylogenetically most closely related to the viviparous clade. However, the biarmed W chromosome characterizing the western viviparous populations is an apomorphic character, whereas the uniarmed W chromosome, existing both in the western oviparous populations and in the geographically distant eastern viviparous populations, is a plesiomorphic character. This suggests an eastern origin of viviparity. Various estimates suggest that the oviparous and viviparous clades of L. vivipara split during the Pleistocene. Our results are discussed in the framework of general evolutionary models: the concept of an oviparity–viviparity continuum in squamates, the cold climate model of selection for viviparity in squamates, and the contraction–expansion of ranges in the Pleistocene resulting in allopatric differentiation.