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May 15,2016
Volume 9,Issue 9
Pages -
Subject Area:Animal Molysmology,Agricultural Entomology,Horticulture,Crop Cultivation,Ecology,Botany

Title: Measurement of logarithmic phase and toxicity in Leptospira

Authors: XIE Xufeng, ZHANG Wenlong, CAO Yongguo
PP: 968-973
Abstract: Objective: To explore the growth characteristics and toxicity of leptospira for the next research. Methods: Leptospira was incubated in EMJH medium using a shaker at 28-30℃ and used bacteria count plate and dark-field microscope to measure the viable count every 6 h. Being chosed 4 groups of 4 to 6-week-old female golden hamsters, each group having four hamsters. Intraperitoneal challenge of hamster with different concentrations’ leptospiroses, keeping a record of the days these hamsters died in half. Besides, being chosed 3 groups of hamsters, intraperitoneally challenged 106 leptospiroses living in logarithmic phase, measuring the lethal dose and geted the infected hamsters’ kidneys, livers and lungs to do histopathological examination. Results: 1) During 0-150 h, Leptospira growth showed S curve. During 0 to 60 h, it growed slowly, developed rapidly in 80-105 h but in 130-150 h came into stable phase. 2) Intraperitoneally challenging hamsters of 102-105 Leptospira all died an half but 3 groups of hamsters intraperitoneally challenged 106 all died in 5-7 d. 3) Histopathological examination showed that infected hamsters’ lungs bled obviously, livers’ cells tight coupling broadened, kidney tubules also bled. Conclusion: The logarithmic phase of Leptospira was 80-105 h in EMJH medium at 28-30℃. The medium lethal dose of Leptospira in 4 to 6-week-old female golden hamster was 102, lethal dose was 106. Infected hamsters showed obviously pathological damage.
Keywords: animal molysmology; Leptospira; logarithmic phase; LD50; pathology

Title: Establishment of Cj0448c mutant in Campylobacter jejuni

Authors: HAO Haihong, HU Bo, LIU Zhenli, YUAN Zonghui
PP: 961-967
Abstract: Campylobacter jejuni is one of a leading zoonotic pathogen. The antibiotic resistance of this pathogen has raised the world concern. In the previous study, the microarray assay indicated that Cj0448c was down-regulated in the intermediate-to-high level of macrolide resistant C. jejuni. The aim of this study was to investigate the role of this gene on the resistance and virulence. In the present study, C. jejuni NCTC1168 was used as the reference strain, the macrolide resistant mutant was obtained by exposure of the reference strain to increased concentration of erythromycin. The Cj0448c deletion mutant was constructed via insertional inactivation. The result of this study can provide a basis for further study on the function and regulation mechanism of this gene.
Keywords: veterinary pharmacology; Campylobacter jejuni; antibiotics resistance; Cj0448c; gene knock out

Title: The detection of integrin α4β7 in mastitis cow

Authors: QU Shihui, WANG Haixia, LIU Hui, LIU Wenbo, ZHANG Naisheng
PP: 955-960
Abstract: Bovine mastitis is one of the major diseases which cause serious economic loss to the dairy farming worldwide. Mammary gland is an important organ to the cow, the lymphocyte can not only fight against pathogens, but also provide assistance from a passive immunization children through breast milk. Research has shown that the migration of lymphocyte has tissue specificity, which depends on the expression of adhesion molecules on the vascular endothelium and cell surface. To further research the differential expression of lymphocytes homing receptor α4β7 in healthy and mastitis bovine, we took blood and milk from healthy and mastitis bovine, respectively, and used flow cytometry to detect lymphocytes which positive express α4β7. The results showed that α4β7 was expressed on lymphocytes both in blood and milk from healthy and mastitis bovine, and the positive rate in mastitis bovine was higher than the healthy bovine.
Keywords: clinical veterinary medicine; mastitis; lymphocytes; α4β7

Title: Effects of ascorbic acid on germination and seedling growth in Elymus sibiricus seeds with different ageing duration

Authors: YAN Huifang, ZHU Yanqiao, MAO Peisheng
PP: 949-954
Abstract: For purpose to explore the effects of ascorbic acid (AsA) on germination and seedling growth of aged Elymus sibiricus seeds, the present study measured the germination percentage, shoot length, shoot weight, root length, root weight and vigour index of AsA pre-soaked E. sibiricus seeds which were accelerated ageing for 12, 36, 60, 84, 108 h at 45℃ and 100% relative humidity conditions. The results indicate that AsA significantly (P＜0.05) inhibits the loss of germination percentage, shoot length, root weight and vigour index of aged seeds at low ageing level (below 36 h), whereas has no significant effects (P＞0.05) on seed germination and seedling growth at high ageing level (60-108 h). There is a close relationship between the effects of AsA on seed germination and seedling growth and the degree of seed ageing.
Keywords: range science; Elymus sibiricus; ascorbic acid; germination; seedling growth; different ageing duration

Title: Genetic variation in six orchardgrass (Dactylis glomerata L.) cultivars based on microsatellite analysis

Authors: JIANG Linfeng
PP: 942-948
Abstract: In this study, microsatellite analysis was employed to investigate genetic variation and relationships among and within six orchardgrass cultivars. Results indicated that: 1) The average number of genetic distance among six Dactylis glomerata L. cultivars was 0.143 7, below 0.15, and 65.30% of the genetic diversity existing within analysis cultivars, while genetic difference among cultivars was 34.70%. 2) 114 clear bands were amplified by 15 microsatellite primers, based on 150 individual plants from six D. glomerata L. cultivars, the average number of amplified bands by each pair of primer was 7.6, and the percentage of polymorphic loci was 100%. Polymorphism information content (PIC) ranged from 0.4 to 0.5 with an average of 0.46. These results suggested that microsatellite markers had a wide prospective in genetic variation analysis of orchardgrass, and the results had important reference values for D. glomerata L. cultivars.
Keywords: range science; Dactylis glomerata L.; cultivars; microsatellite; genetic variation

Title: Detection of phage WO and Wolbachia in Corcyra cephalonica and both distribution in different organs

Authors: CONG Bin, ZHANG Mingzhu, LI Shuaiqiang, ZHANG Tongshu, DONG Hui
PP: 933-941
Abstract: Objective: Many Wolbachia strains were found infected by phage WO. It is postulated that phage WO involves in the control of genetic variation of Wolbachia and the regulation of reproduction of the hosts. To further understand the interactions between phage WO, Wolbachia and the hosts, laboratory tests were conducted to detect the presence of phage WO and Wolbachia in different organs of Corcyra cephalonica. Methods: Total DNAs of different organs and the adult C. cephalonica were extracted using SDS-proteinase K method; PCR technique was used to detect the infection of phage WO and Wolbachia with primers of Wolbachia surface protein gene (wsp) and putative phage capsid protein gene (orf 7). Furthermore, the distributions of phage WO and Wolbachia in head, thorax, abdomen, leg and ala were also analyzed. Results: For the whole body of C. cephalonica, wsp sequence was specifically amplified as 602 bp; the orf 7 primer amplified two sequences (409 bp and 424 bp) in total DNA of the whole body. In addition, PCR with these two primers also successfully amplified their target fragments in different organs. Conclusion: The head, thorax, abdomen, leg and ala of C. cephalonica were all infected by Wolbachia and phage WO. The phylogenetic tree constructed with wsp gene showed that all Wolbachia in C. cephalonica belong to Pip subgroup; Blasting search analysis showed that Wolbachia in C. cephalonica were infected by two kinds of phage WO, which were WOcep1 (409 bp) and WOcep2 (424 bp), respectively.
Keywords: agricultural entomology; Corcyra cephalonica; Wolbachia; phage WO; wsp; orf 7

Title: Cloning and bioinformatics analysis of terpene glucosyltransferase in Vitis vinifera L. ‘Gewurztraminer’

Authors: WEN Yaqin, LI Xiangyi, PAN Qiuhong
PP: 925-932
Abstract: This study aimed at the cloning of eight putative terpene glycosyltransferase genes (UGTs) from the berries of Vitis vinifera L. ‘Gewurztraminer’ and the bioinformatics analysis of these genes. The results showed that the eight UGTs all possessed complete open reading frame and glycosyltransferase-based conservative amino acid area PSPG primitives that could be translated to produce the complete proteins. Evolutionary tree analysis revealed that the sequence similarity between these genes reached 63.2%-76.1%, and the homology of these genes with Arabidopsis glucosyltransferases was within the range of 49.4%-59.3%. This work lays a foundation for subsequent function identification of these genes.
Keywords: horticulture; terpene glucosyltransferase; gene sequence; grape berry

Title: Heredity of ornamental characters in F1 hybrid generation of cut-flower chrysanthemums

Authors: LI Fangyuan, YANG Yunyan, WEN Chao, LIU Meijuan, ZHAO Liangjun
PP: 916-924
Abstract: Heredity of some characters in F1 generation by the eight cross combinations among eight varieties of cut-flower chrysanthemums were studied. Eight main ornamental characteristics of the F1 hybrids such as flower color, flowering date, flower diameter, ray floret number, tubular flower number, length of petal, petal width, pedicel length were analyzed. The results show that different flower colors have different genetic potential, and each color of the parents in turn can be ranked as follows: white > pink > yellow; the flower time of F1 hybrids lied between parents, but the early flowering occurs. The mean of hybrid inflorescence diameter, the number of ligulate flower and tubular flower, petal length and width, all showed a trend of decrease and the means of them were 95.9%, 85.2%, 71.5%, 76.1%, 84.9% of their parents, respectively. But heterosis in different cross combinations were not the same, and the performance advantage depended on combinations. The means of hybrid pedicel length had a clear difference from their parents and showed a trend of decrease. According to breeding objectives, having a significant correlation among different ornamental characters could provide data reference of cut-flower chrysanthemums.
Keywords: hoticulture; cut-flower chrysanthemums; hybrid; ornamental character; heredity

Title: Studies on the effects of controlled loss and conventional fertilizer on growth and development and yield of summer maize

Authors: JIANG Qian, LI Lei, ZHANG Fenglu, WANG Juhui, YUAN Liuliu, WU Ruijuan
PP: 909-915
Abstract: The effects of controlled loss and ordinary fertilizer on dry matter accumulation and partitioning, leaf areas, leaf photosynthetic rate, stalk characters and grain yield were studied using maize hybrid Zhengdan 958 as material. The results showed that with the same nutrient amount (N 144 kg/hm2, P2O5 72 kg/hm2, K2O 72 kg/hm2) of controlled-loss and ordinary fertilizer applied, there were significant difference in growth and development, dry matter accumulation and partitioning, and maize yield. The effect of controlled loss fertilizer treatment was significantly better than that of ordinary fertilizer. Compared with ordinary fertilizer, the 1 000 grain weight, dry matter and yield of maize of controlled loss fertilizer could be increased by 12.3%, 12.8% and 6.9%, respectively. The leaf area index and soil and plant analyzer develotrnent (SPAD) value were increased significantly during grain filing stage and the photosynthetic rate increased 26.5% at dough stage for controlled loss fertilizer treatment. At the same time, the basal inter-node thickness and fracture resistance were increased, so the loading resistance ability was enhanced. As a new type of fertilizer, controlled loss fertilizer will play an important role in improving the yield of maize, increasing the efficiency of fertilization and protecting the environment.
Keywords: crop cultivation; maize; controlled loss fertilizer; dry matter accumulation and partitioning; yield; plant morphology

Title: Influence of altitude on reproductive strategies of Halenia elliptica

Authors: WANG Shasha, ZHANG Renyi, ZHENG Kai, FENG Shuo, YUAN Jianli
PP: 901-908
Abstract: Halenia elliptica is a common species in the eastern Tibetan plateau. We sampled H. elliptica along with elevation, and aimed to study its individual size and reproductive strategies in various altitude gradients and examined how environmental factors contribute to species evolution. Results showed: 1) Individual size decreased, while reproductive allocation and reproductive export increased along with elevation. When the effect of individual size was controlled statistically, significant positive correlation was found between elevation and reproductive allocation, as well as elevation and reproductive export. 2) Reproductive allocation decreased with individual size increasing, but there was no significant relationship between reproductive export and individual size. When the effect of elevation was controlled statistically, reproductive allocation was negatively related to individual size, whereas no significant relationship between reproductive export and individual size was found. 3) Individual size within the population was more centrally distributed as elevation increasing. As an environmental factor, altitude has a great influence on individual size, reproductive allocation and reproductive export.
Keywords: ecology; altitude; reproductive allocation; reproductive export; individual size; correlation

Title: Study on seed dormancy mechanism and breaking technique of Halenia elliptica

Authors: CHEN Rongjian, FENG Shuo, YU Hongqiang, WANG Junwu, YUAN Jianli
PP: 893-900
Abstract: The germination rate of Halenia elliptica was low, and the degree of dormance was deep. The seed dormancy mechanism was revealed, and the best solution of breaking seed dormancy was explored by determining seed viability, water absorption and germination rate of seeds under different treatments. The results showed that: 1) The activity of H. elliptica seed was high, the life force of the seed can be reached (85.33+2.49)% by triphenyl tetrazolium chloride (TTC). 2) The water absorption rates of broken and intact seeds were different in the early 14 h, but the ultimate water absorption rates were no difference, which reached (92.64±0.88)% and (92.95±0.82)% respectively, suggestting that seed coat had no effect on seed water absorption. The germination rate of H. elliptica seed was significantly increased after NaOH treatment (P<0.05), which suggested that the seed capsule barrier was one of factors of the seed dormance. The seed germination rate was significantly improved by NaOH and GA treatment (P<0.01), which suggested that the intrinsic inhibitors was another factor of seed dormance. So it was considered that the dormance of H. elliptica seed was the comprehensive dormance of the seed coat and inhibitors contained in the seeds. 3) The final germination rates of the seeds treated by different methods were significantly improved, among them, seeds soaked with water by 2 d can make the coat swell, constantly absorbing the water needed for germination; seeds soaked with 30% NaOH by 80 min and then imposed 200 μg/g GA, the effect was the best when the temperature was 25℃, germination rate reached (74.80%±1.71)%.
Keywords: ecology; Halenia elliptica; dormancy mechanism; germination rate; dormancy breaking

Title: Molecular characterization and evolutionary analysis of b-type Farinin genes in Brachypodium distachyon L.

Authors: SUBBURAJ Saminathan, ZHU Gengrui, LI Xiaohui, HU Yingkao, YAN Yueming
PP: 887-892
Abstract: Farinin is one of the oldest members of the gluten family in wheat and Aegilops species, and related to dough properties. Here, we performed the detailed molecular genetic study on Farinin genes in Brachypodium distachyon L. A total of 51 b-type Farinin genes were cloned and characterized from 14 different B. distachyon L. accessions by allele-specific PCR (AS-PCR). All genes were highly similar to those previously reported from wheat and Aegilops species. The identification of deduced amino acid sequences showed that b-type Farinins across Triticeae genomes could be classified as b1-, b2-, b3-, and b4-type Farinin, while B. distachyon L. had only b3- and b4-type Farinin. Phylogenetic analysis suggested that Brachypodium Farinin may have closer relationships with common wheat and further confirmed four different types of b-type Farinin present in Triticeae and Brachypodium genomes, corresponding to b1, b2, b3 (Group 1), and b4 (Group 2). The b4-type could be considered older in the Farinin family.
Keywords: botany; Brachypodium distachyon L.; Farinin; molecular cloning; evolution

Title: An improved oocyte retrieval after superovulation on rhesus and cynomolgus monkeys

Authors: GAO Jiahong, HUANG Zhangqiong, JIANG Qinfang, ZENG Qiao, WU Zhengcun
PP: 881-886
Abstract: Objective: To explore an improved, high efficient and mild hurt operation technique for oocyte retrieval in rhesus and cynomolgus monkeys. Methods: To locate and measure ovum with B-type ultrasonography after superovulation in rhesus and cynomolgus monkeys. We catheterized with self-made catheter to relieve occulusion of bladder, and then we operated an 1.5-2.5 cm incision in the center of hypogastrium or two incisions of two hypogastrium sides, respectively. The ovary was exposured, and then aspirated ovarian follicles under negative pressure with a 5 mL syringe which attached 23 Gauge needle. Results: 11 rhesus monkeys and 13 cynomolgus monkeys had been undergone this improved operation. The average size of ovum was 2.2 cm×2.4 cm in rhesus monkeys and 2.1 cm×2.4 cm in cynomolgus monkeys. The average size of incisions was (2.3±0.2) cm in rhesus monkeys and (2.2±0.1) cm in cynomolgus monkeys. The total average operation time was (44±10) min. The average number of oocyte retrieval was 26.5±15.8 in rhesus monkeys and 25.1±16.3 in cynomolgus monkeys. Conclusions: It is a feasible, high efficient and low hurt way that we cut a small incision for oocyte retrieval after catheterization through locating and measuring ovum with B-type ultrasonography.
Keywords: zoology; rhesus monkey; cynomolgus monkey; superovalution; B-type ultrasonography; urethral catheterization

Title: A new discovery of the basic human auricle formation process

Authors: LU Xiangyue
PP: 875-880
Abstract: Textbooks think: human auricle is from 6 branchial hillocks which around the first branchial cleft. Condition of auricle development show that auricle of human embryos grow not just from the 6 branchial hillocks samples, also from branchial hillocks under embossments. In the end, the embossments make up most of the auricle.
Keywords: physiology; embryo; auricle; reflecting region; auricular point

Title: Evaluation of SUMO-and Strep-tagged TRAIL protein prepared by two step of purification method

Authors: HAN Xiaojian, HUANG Jingjing, HAO Zhichao, JIN Aishun
PP: 868-874
Abstract: In order to obtain tumor necrosis factor related apoptosis-inducing ligand (TRAIL) protein with the high purity and biological activity, a two-step purification method was prepared using SUMO and Strep tagged, and the purity and activity of TRAIL were evaluated. Firstly, pSUMO-Strep-hTRAIL plasmid was constructed and transfected into prokaryotic cells to produce bacterial fluid, and then TRAIL protein of high purification was obtained by Ni+ and Strep using a two-step affinity chromatography. In order to evaluate the biological activity of TRAIL, the effect of TRAIL on inducing apoptosis was detected. The results showed that the TRAIL protein prepared by established method has good biological activity, which provides the material for anti-tumor experiments in vivo.
Keywords: biotechnology; purification method; biological activity; tumor necrosis factor related apoptosis-inducing ligand

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