The use of Verticillium dahliae and Diplodia scrobiculata to induce resistance in Pinus halepensis against Diplodia pinea infection

The pathogenic fungi Verticillium alboatrum and Diplodia scrobiculata were assayed for biological control of Diplodia pinea on Aleppo pine (Pinus halepensis) in Catalonia (north-eastern Spain). Young shoots were pre-treated with inoculations of either V. dahliae or D. scrobiculata, by placing colonized agar plugs on wounds made by removing a single needle fascicle. An inoculation with D. pinea was performed 15 days later. Two months after the shoot inoculations, the canker length on the stems was measured and the percentage of shoot dieback calculated. Verticillium dahliae and D. scrobiculata were found to significantly reduce the canker length of D. pinea (P < 0.05) when compared with positive controls. Diplodia pinea was slightly more sensitive to V. dahliae than to D. scrobiculata, but no significant differences (P > 0.05) were observed in the mean canker length between the two treatments. Trees pre-inoculated with V. dahliae resulted in 31.12% shoot dieback, while those pre-inoculated with D. scrobiculata resulted in 32.18% shoot dieback, compared with positive controls (42.85%).     

Variable response of open-pollinated seedling progeny of avocado to Phytophthora root rot

Phytophthora root rot, caused byPhytophthora cinnamomi Rands, is the most important disease of avocado (Persea americana Miller). In an attempt to identify root rot-resistant rootstocks that could ultimately be used under conditions in southern Florida, we screened open-pollinated progeny of avocado from the National Germplasm Repository in Miami. From 1996 to 1998, a total of 2,355 seedlings from 51 accessions were examined in potting mix artificially infested withP. cinnamomi. Most seedlings developed severe root rot, but tolerance was observed in some families (i.e., progeny of certain accessions). Although the most susceptible families developed mean disease ratings of up to 97% root necrosis, mean ratings for the most tolerant families were less than 60%. There was also a strong relationship between the racial background of the female parent and the tolerance of seedlings. Seedlings of the West Indian race and hybrids between it and the Guatemalan race were significantly more tolerant than those from other parents (P< 0.05). Individuals in several families developed < 50% root necrosis, the arbitrary standard of tolerance in this study. Twelve families accounted for 82% (188 of 229) of the tolerant seedlings, and only two of these did not have a West Indian or Guatemalan × West Indian pedigree. Broad-sense heritability for PRR tolerance was 0.45. This is the first report on the inheritance of PRR tolerance in avocado and on the influence of genotype and racial pedigree under controlled conditions.     

Enzymatic profiles of Erysipelothrix rhusiopathiae and Erysipelothrix tonsillae

The enzymatic activities of 39 strains of Erysipelothrix rhusiopathiae and 34 of E tonsillae were determined with the API ZYM system. The profiles of these two species were very similar, differing solely in N-acetyl-beta-glucosaminidase activity. Whereas 90 per cent of strains of E rhusiopathiae exhibited strong activity with N-acetyl-beta-glucosaminidase, positive reactions were observed for this enzyme in only 24 per cent of strains of E tonsillae. These results support previous DNA-DNA hybridisation studies and suggest that E tonsillae is a new species of the genus Erysipelothrix.     

Induction of oestrus by administering Inhibin antiserum along with equine chorionic gonadotropin in anoestrous bitches

As dogs experience oestrus only once or twice a year, it is necessary to establish an effective method of oestrous induction for efficient breeding. In the present study, we evaluated inhibin antiserum (IAS) on oestrous induction in anoestrous females. Bitches were administered 0.5 ml/kg IAS or a mixture of 50 IU/kg equine chorionic gonadotropin (eCG) and 0.5 ml/kg IAS and 500 IU human chorionic gonadotropin (hCG) administered 7 days after the mixture injection. As a control, bitches received 50 IU/kg eCG, with 500 IU hCG administered 7 days after eCG injection. Blood-tinged vaginal discharge, vulvar swelling, plasma progesterone concentrations and ovarian follicular development were assessed from day 0 to day 14. IAS alone injection did not induce oestrus in bitches at the anoestrous stage. Conversely, vulvar swelling, blood-tinged vaginal discharge and an estimated luteinizing hormone (LH) surge appeared on days 3–7, days 3–6 and days 7–9 after the IAS+eCG mixture injection, respectively, in all five bitches at the anoestrous stage. The average number of developing and ovulated follicles in bitches administered IAS+eCG was 8.8 and 9.6 respectively. A single eCG injection followed by hCG induced oestrous signs, with an average of 8.3 developing follicles and 4.5 ovulated follicles. This study revealed that IAS alone did not induce oestrus, but when IAS was used in combination with eCG, it induced oestrus and promoted a considerable number of ovulations in anoestrous dogs.     

Changes in the expression of toll-like receptor mRNAs during follicular growth and in response to lipopolysaccharide in the ovarian follicles of laying hens

The aim of this study was to determine the changes in the mRNA expression of Toll-like receptors (TLRs) in hen ovarian follicles during follicular growth and in response to lipopolysaccharide (LPS). White follicles and the fifth largest to largest follicles (WF and F(5)-F(1), respectively) were collected from laying hens. To examine the effects of LPS, the laying hens were treated intravenously with LPS (1 mg/kg BW) 0, 3, 6, 12 and 24 h before examination. Expressions of TLRs and IL-1beta in the theca and granulosa layers were examined by semi-quantitative RT-PCR. Immunocytochemistry was performed to identify immunoreactive TLR-4. The theca layer expressed TLR-2, TLR-4, TLR-5 and TLR-7, whereas the granulosa layer expressed only TLR-4 and TLR-5. The expression of TLR-4 and TLR-5 in the theca layer increased significantly during follicular growth. In the granulosa layer, the expression of TLR-5 increased, but that of TLR-4 was unchanged. Expression of TLR-4 increased significantly during the period of 6 to 12 h of LPS treatment in the theca layer and during the period of 12 to 24 h in the granulosa layer of F(3). Immunoreaction products for TLR-4 were observed in theca interna and granulosa layers of WF and F(5)-F(1), with the greater amount observed in the theca interna. LPS treatment significantly increased expression of IL-1beta in the theca layer after 3 h and in the granulosa layer during the period of 12 to 24 h. These results suggest that TLRs are expressed in ovarian follicles and that TLR-4 and TLR-5 expression increase with the growth of follicles. Enhanced expression of TLR-4 and IL-1beta by LPS in the theca and granulosa layers suggests possible roles of TLR in recognition of microorganisms.     

Effect of ovary lipid of skipjack tuna on rat serum components after stress application

Lipid extracted from the ovary of skipjack tuna by the method that we developed is rich in phospholipid-type docosahexaenoic acid. The ovary lipid of skipjack tuna (OLS) was studied for its anti-stress activity in male Wistar rats, focusing on stress-related blood components: recovery from stress was examined after application of water immersion restraint stress. As a result, serum corticosterone (CORT) secretion was inhibited and decreased rapidly after stress application in rats given OLS compared with control rats. As CORT acts as a glucocorticoid, non-esterified fatty acid (NEFA) is expected to increase by stress application. However, the concentration tended to be lower in rats given OLS than in control rats. With respect to OLS concentration, OLS increased serum dehydroepiandrosterone, secretion concentration-dependently. In addition, as with the recovery study, it tended to inhibit the increase in NEFA. These results indicate that OLS may have an anti-stress activity against acute stress.     

大豆抗菌核病的全基因组关联研究

找到大豆与抗菌核病强关联的候选位点或候选基因,为抗病基因克隆和抗病分子标记开发提供借鉴,服务大豆抗菌核病育种。对126个加拿大大豆品种的基因组DNA用ApekⅠ酶消化后Illumina Hiseq2000平台测序进行基因分型,供试的126个材料用棉垫接种核盘菌菌丝体进行表型鉴定。采用Structure 2.3.4、SPSS 20.0、TASSEL 5.0和PLINKv 1.07软件分别模拟群体遗传结构、二元主成分分析、邻接法聚类,进行SNP-phenotype和Haplotype-phenotype的关联分析(只考虑加性效应)。最小等位基因频率0.01过滤,得到30 125个SNPs。主成分及群体结构聚类结果中度一致,将126个供试材料划分为2个组群,Kappa聚类一致度检验K=0.44。邻接法(The neighbor-joining algorithm,NJ)聚为3个组群。α≤0.05时,在单个SNP-phenotype的关联研究中,最强关联在3号染色体物理位置34387780、34387823和34387841处(P值都为8.669E-7),可分别解释表型变异的17.80%,其次在20,1,4,17号染色体上。Haplotype-phenotype的关联分析中,最强关联在17号染色体物理位置5575883/5647814/5648648/5734897处(P值为1.038E-6),可解释表型变异的17.56%。200 kb范围内,3号染色体上的候选基因有Glma.03g129100、Glma.03g129200、Glma.03g129300、Glma.03g129500、Glma.03g129800、Glma.03g129900。17号染色体上为Glma.17g071300、Glma.17g072200、Glma.17g073300。     

Identification of ommastrephid squid paralarvae collected in northern Hawaiian waters and phylogenetic implications for the family Ommastrephidae using mtDNA analysis

ABSTRACT:   A total of 110 adult individuals from four ommastrephid (family Ommastrephidae) squid species ( Ommastrephes bartramii, Sthenoteuthis oualaniensis, Eucleoteuthis luminosa, and Hyaloteuthis pelagica ) were used to obtain diagnostic DNA markers for species identification. Restriction fragment length polymorphism (RFLP) analysis of a partial segment (855 bp) of the mitochondrial DNA cytochrome oxidase I (COI) gene amplified by polymerase chain reaction (PCR) revealed that the restriction profiles of two endonucleases ( Alu  I and Tsp 509 I) were diagnostic for species identification. The restriction assay partially supplemented with nucleotide sequence analysis successfully assigned 69 damaged and morphologically equivocal ommastrephid paralarvae collected in northern Hawaiian waters, identifying 60 O. bartramii , eight S. oualaniensis , and one E. luminosa . The family Ommastrephidae appears to be monophyletic. Although the phylogenetic relationships among genera were not resolved well due to apparent homoplasy and large genetic divergence between species, COI sequence data without transitions provided support for subfamily level relationships.     

Lethal effects caused by suspended particles and sediment load on zoospores and gametophytes of the brown alga <Emphasis Type="Italic">Eisenia bicyclis</Emphasis>

ABSTRACT:   The effects caused by suspended particles in seawater and sediment cover of the substrate on the attachment to the substrate of zoospores, and on the subsequent growth and survival of gametophytes of the brown alga Eisenia bicyclis were examined in the laboratory. The attachment rate to the substrate of the zoospores was remarkably reduced by the sediment cover on the substrate, and it decreased to 3.8% of the no-sediment control by a slight sediment cover of 3.0 mg/cm² (0.048 mm in thickness). The growth of gametophytes was not inhibited at 10 mg/cm² or less of sediment cover, though it was stopped at 30 mg/cm². The survival rate of gametophytes became 39.4% at 5 mg/cm² on the substrate, and gametophytes were all blighted at 30 mg/cm². The ratio of male to female of the surviving gametophytes became 81.5:18.5, when the sediment cover was 10 mg/cm². The rate of total loss ( TL [%]) of zoospores and gametophytes of Eisenia bicyclis was obtained from the following equation: TL  = 100 (1 − exp[−0.0339  C ] exp[−1.24  Q ]), where, C (mg/L) and Q (mg/cm²) indicate the concentration of suspended particles in seawater and the sediment on the substrate, respectively.     

The effects of vitamin E on non-specific immune response of the juvenile soft-shelled turtle <Emphasis Type="Italic">Pelodiscus sinensis</Emphasis>

ABSTRACT:   This study was conducted to determine the effect of dietary vitamin E supplements on non-specific immune responses in juvenile soft-shelled turtles. Turtles were fed diets supplemented with vitamin E at 0 (control), 50, 250, 500, 1000 and 5000 mg/kg, respectively, for 4 weeks. Results showed that blood cell phagocytosis and serum bactericidal activity were significantly improved in turtles from two diets (250 and 500 mg/kg vitamin E supplementation), while had no significant improvement in turtles from the three remaining diets (50, 1000 and 5000 mg/kg vitamin E supplementation) when compared to turtles from the control group. Serum bacteriolytic activity in turtles from diets with 50, 250, 500 and 1000 mg/kg vitamin E supplementation were higher than activity levels quantified for the control group, but no significant difference was observed between the 5000 mg/kg vitamin E supplementation diet and the control diet. The results suggest that vitamin E has an upper and lower threshold for improving non-specific immune function in soft-shelled turtles, and the optimal supplementation may be between 250 and 500 mg/kg.