新疆春小麦育成品种遗传演变分析

【目的】研究新疆春小麦育成品种遗传演变规律,提高小麦资源的利用效率,为新疆春小麦品种选育和改良提供参考依据。【方法】以新疆春小麦育成品种为材料,对其主要性状进行综合评价分析,以春小麦90K芯片开展新疆春小麦育成种亲缘关系分析。【结果】新疆春小麦育成种遗传多样性丰富,平均遗传多样性指数2.005,变幅为1.902~2.181。相关性分析结果表明,蛋白质含量、湿面筋含量、穗粒数与育种年代呈极显著正相关。新疆春小麦品种选育主要是通过常规杂交育种、杂交辐射诱变育种、引种3种方式。利用小麦90K芯片将新疆春小麦育成种划分为3个类群,显示了各品种之间的亲缘关系。【结论】新疆春小麦育种遗传基础薄弱、遗传多样性逐渐散失,新疆小麦育种应加强资源收集与利用,扩大育种亲本选择,提高品种变异的丰度和广度,以多抗、高产稳产、优质高效的聚合育种,加快新品种选育进程,提高育种效率。     

Development of a method for live octopus (Octopus vulgaris) transportation for long distance at high densities

The development of new octopus‐based products with a growing economic value ensures the commercial interest of this species, making live octopus export an activity of great interest. The aim of this study was to develop a method for long‐distance transportation of live octopus at high densities. The system was composed by 220‐L tanks with cooling and aeration, where the animals were kept separated from each others. The water temperature was maintained at 10°C, after a decreasing of 1°C/hr. Live octopus transportation was tested for 48 hr at two densities: 50 kg/m³ and 100 kg/m³. During this period, water parameters were monitored. Stress response was evaluated through the analysis of haemolymph, muscle and brain tissues. No mortality was registered after 48 hr for both treatments. In all trials, water quality remained within the normal limits in both densities; there was, however, a significative increase of ammonia levels in the water. Ammonia, dopamine and Hsp70 levels were analysed in the beginning and at the end of the experiment for both densities; however, no significant differences were found among them. In general, this system seems to be a viable solution for live octopus 48‐hr transportation at a density of 100 kg/m³.     

文心兰HSP70基因的克隆及表达分析

为研究文心兰热激蛋白70基因（命名为OnHSP70）的分子特性及表达特点,以文心兰‘柠檬绿’（Oncidium hybridum ‘Honey Angel’）为材料,采用RT-PCR技术克隆OnHSP70,利用生物信息学方法分析其分子特性,通过qRT-PCR技术分析其在不同组织及不同非生物胁迫处理下的表达特点。生物信息学分析表明：该基因开放阅读框为1944 bp,编码647个氨基酸,翻译的蛋白为稳定蛋白,属于HSP70超家族,其蛋白二级结构由41.11%的α-螺旋、17.77%的延伸链、7.73%的β-转角和33.38%的无规卷曲组成,具有2个高度保守的功能域。聚类分析表明：该蛋白与铁皮石斛和深圳拟兰的HSP70亲缘关系较近。qRT-PCR分析表明：文心兰HSP70在4种不同组织器官中具有表达差异性,在花中的表达量最高;高温处理后基因的表达量明显上升;40 ℃高温胁迫4 h时表达量达峰值;茉莉酸甲酯及水杨酸处理时表达量呈现下调响应。本研究为后期该基因功能研究及提高文心兰对高温的适应性提供理论基础。     

Prevalence,geographic distribution and phenotypic differences of Piscirickettsia salmonis EM‐90‐like isolates

Early reports accounted for two main genotypes of Piscirickettsia salmonis, a fish pathogen and causative agent of piscirickettsiosis, placing the single isolate EM‐90 apart from the prototypic LF‐89 and related isolates. In this study, we provide evidence that, contrary to what has been supposed, the EM‐90‐like isolates are highly prevalent and disseminated across Chilean marine farms. Molecular analysis of 507 P. salmonis field isolates derived from main rearing areas, diverse hosts and collected over 6 years, revealed that nearly 50% of the entire collection were indeed typed as EM‐90‐like. Interestingly, these isolates showed a marked host preference, being recovered exclusively from Atlantic salmon (Salmo salar) samples. Although both strains produce undistinguishable pathological outcomes, differences regarding growth kinetics and susceptibility to the antibiotics and bactericidal action of serum could be identified. In sum, our results allow to conclude that the EM‐90‐like isolates represent an epidemiologically relevant group in the current situation of piscirickettsiosis. Based on the consistency between genotype and phenotype exhibited by this strain, we point out the need for genotypic studies that may be as important for the Chilean salmon industry as the continuous surveillance of antimicrobial susceptibility patterns.     

EIAV Rev蛋白拮抗eqTRIM5α介导的AP-1信号通路的机制研究

为探究马传染性贫血病毒(EIAV)附属蛋白Rev负调控Tripartite motif-containing protein 5α(TRIM5α)介导的AP-1信号通路的机制,本研究将pEIAV-Rev-HA和pcDNA3.1质粒分别与含TRIM5α基因的质粒及pGL3-AP-1-Luc(AP-1报告质粒)共转染HEK 293T细胞,采用荧光素酶试验检测Rev对TRIM5α激活的AP-1信号通路的影响;将pEIAV-Rev-HA和pcDNA3.1质粒分别与含TAK1、TAB2、P38和c-Jun基因的质粒及pGL3-AP-1-Luc共转染HEK 293T细胞,采用荧光素酶试验检测Rev对TRIM5α下游转导分子(TAK1、TAB2、P38、c-Jun)激活的AP-1信号通路的影响;将pEIAV-Rev-HA和pcDNA3.1质粒分别与含TAK1、TAB2、P38基因的质粒共转染HEK293T细胞,利用western blot试验分别检测TAK1、TAB2、P38的表达水平;将pEIAV-Rev-HA和pcDNA3.1质粒分别与含P38基因的质粒共转染HEK 293T细胞后加入蛋白酶体抑制剂MG132,利用western blot检测P38蛋白的表达情况。结果显示,共转染EIAV-Rev-HA实验组中TRIM5α对AP-1的激活倍数为0.4,而共转染pcDNA3.1对照组中相应的激活倍数为26.0;共转染pEIAV-Rev-HA实验组中,TAK1、TAB2、P38和c-Jun对AP-1信号通路的激活倍数分别为7.7、0.1、0.6、9.8,而共转染pcDNA3.1对照组中对AP-1信号通路的激活倍数分别为60.0、1.5、6.3、12.0;转染pEIAV-Rev-HA+pP38-Flag组与转染pcDNA3.1+pP38-Flag组相比,前者P38蛋白的表达量显著降低;加入蛋白酶体抑制剂组则恢复了P38蛋白的表达。上述结果表明,EIAV Rev显著下调eqTRIM5α及其下游转导分子TAK1、TAB2、P38激活的AP-1信号通路,但不显著下调c-Jun激活的AP-1信号通路;EIAV Rev通过蛋白酶体途径降解P38蛋白的表达而抑制eqTRIM5α激活的AP-1信号通路。本研究结果为理解EIAV与宿主蛋白相互作用提供参考依据。     

Effect of Heat Shock Protein-90 (HSP-90) mRNA Expression in Intestinal Tissuse of Chickens Infected with E.necatrix

The 90 chickens were randomly divided into control group, E.necatrix one time infection group and E.necatrix two times infection group.We used duplex PCR method to test the expression changes of HSP-90 mRNA in intestinal tissue of chickens infected with E.necatrix and researched the effects of it comprehensively and systematically.The results were showed that after 0 to 14 days the expression of HSP-90 mRNA in intestinal tissues of 14-day-old chickens infected with E.necatrix presented a upward trend though it was lower than the control chickens.These chickens were infected with E.necatrix again at 28 days, HSP-90 mRNA expressions of duodenum, jejunum and ileum decreased and then increased.However, it always showed a downward trend in appendix.When 28 days chickens were infected with high-does E.necatrix once, HSP-90 mRNA expressions of duodenum, jejunum and appendix were in a declining curve.While it was increased originally and then decreased in ileum.This study indicated that HSP-90 mRNA expressions in intestinal tissue of chickens infected with E.necatrix were inhibited at first, and the expression quantity was closely related to the degrees of intestinal tissue damage.     

Cloning,Expression of Clostridium perfringens α-toxin Gene and Preparation of its Antisera

One pair of primers had been designed and synthesized based on the α-toxin gene of Clostridium perfringens.The complete α-toxin gene fragment was amplified by polymerase chain reaction (PCR), and then was cloned into pGEM-T Easy vector to construct pGEM-T-α.Digested with EcoRⅠ and Hind Ⅲ, a fragment of 1125 bp was cloned into the expression plasmid vector pET-28a(+).The recombinant plasmid was transformed into the BL21(DE3)plys and induced by 1.0 mmol/L IPTG at 37 ℃.The expression product was found to be 46.1 ku as expected one identified by SDS-PAGE, and confirmed by Western blotting with Clostridium perfringens type A antisera, indicating similar reactivity with native α-toxin.Recombinant α-toxin protein was simultaneously found in culture supernatant, postsonic supertanant and inclusion bodies, most protein was expressed in inclusion bodies, which indicated recombinant α-toxin protein was expressed in the extracellular, periplasm and cytoplasm.Recombinant α-toxin protein in postsonic supertanant could not make mice die, indicating its non-toxicity.Toxin-antitoxin neutralization test showed that antisera of recombinant α-toxin protein were specific to α-toxin.Upon immunization of rabbit with the recombinant α-toxin protein, antisera with high antibody titer neutralizing 100 MLD toxin per 1 mL were prepared.