广东省主要红树林植物DNA条形码评价

为评估DNA条形码对鉴定红树林植物的通用性和有效性,在广东红树林分布区域共计采集红树林植物16科22属23种,共144个样品,并进行DNA条形码测序。结果表明:选择的rbcL、matK和trnH-psbA 3个DNA片段的PCR扩增成功率分别为100%、80.29%±8.49%、99.38%±1.25%。测序成果率最高为rbcL 100%,trnH-psbA次之94.57%±5.06%,matK最低75.04%±6.26%。表明rbcL和trnH-psbA片段在红树林群落中都具有较好的通用性。应用BLAST和NJ Tree两种方法计算红树植物的物种识别率。BLAST结果表明,单片段中trnH-psbA的物种识别率最高,为84.48%±12.09%,rbcL次之,matK最低。NJ Tree分析显示单片段中rbcL的物种识别率最高,为66.65%±17.35%;trnH-psbA片段次之,matK片段最低。两张分析方法都显示多个片段组合使用时,rbcL或trnH-psbA是提高物种平均识别率的主要片段。利用单片段rbcL即可获得平均节点支持率最高的红树植物系统发育树,且能准确区分不同树种。trnH-psbA片段可以识别rbcL片段不能识别的物种,可以作为补充片段。综合比较,推荐rbcL、trnH-psbA作为红树林植物DNA条形码片段。     

一株高雄山虫草的分子鉴定

[目的]利用生物信息学方法初步探讨虫草属日本拟青霉的分类地位。[方法]提取日本拟青霉的基因组,并利用通用引物ITSL和ITS4采用PCR技术扩增rDNA18S-28S ITS序列。采用Clustalx1.83和MEGA5软件构建系统发育树,生物信息学比较分析确定其分类地位。[结果]PCR扩增产物为603 bp。将日本拟青霉重新命名为雪花高雄山虫草(Cordyceps takaomontana strain JLsnow)。将虫草重新分类,并对该类群及其特征进行了描述。[结论]从分子水平上说明日本拟青霉与其他虫草之间的差异,该研究为虫草属真菌的分类提供新思路。     

陕西不同地区马铃薯腐烂茎线虫的分离鉴定及同源性分析

旨在鉴定陕西不同地区马铃薯腐烂茎线虫生物型,为其防治提供理论依据。对陕西不同地区马铃薯腐烂茎线虫进行分离,对其形态进行观察并测量特征值,通过茎线虫线粒体COⅠ通用引物、ITS区通用引物及A、 B生物型的特异性引物对不同地区采集马铃薯腐烂茎线虫进行PCR检测及序列分析,利用MEGA6.0进行系统发育树构建及种群同源性分析。结果表明,陕北地区腐烂茎线虫的生物型为B型,关中地区为A型。形态特征值数据显示雌成虫体长、体宽均大于雄成虫,A型的体长、体宽均大于B型。系统发育树结果显示,陕北不同地区的B型马铃薯腐烂茎线虫聚为一支,合阳、兴平的A型马铃薯腐烂茎线虫聚为一支,与分子鉴定结果相一致。     

Prevalence,molecular characterization and risk factor analysis of Ehrlichia canis and Anaplasma platys in domestic dogs from Paraguay

This is the first study to investigate the prevalence and risk factors associated with Ehrlichia canis and Anaplasma platys positivity in dogs from Paraguay. Conventional PCR assays for the E. canis 16SrRNA gene and A. platys p44 gene were carried out in blood samples from 384 dogs from Asunción city, Paraguay. Sequencing and phylogenetic analysis were performed in selected positive E. canis and (16SrRNA gene) and A. platys (16S and p44 genes) samples. The overall prevalence of E. canis and A. platys in dogs in Paraguay was 10.41% (40/384) and 10.67% (41/384), respectively. Older dogs without veterinary care had higher odds for E. canis positivity and a higher number of dogs in the same household, as well as absence of anti-tick treatment were considered risk factors for A. platys. Ehrlichia canis and A. platys circulate in the dog population from Asunción, and are described for the first time in Paraguay.     

Antibiotic resistance,ESBL genes,integrons, phylogenetic groups and MLVA profiles of Escherichia coli pathotypes isolated from patients with diarrhea and farm animals in south-east of Iran

The aims of this study were to investigate the prevalence, antibiotic resistance, presence of class 1 and 2 integrons, Extended Spectrum β-Lactamases (ESBL) genes, phylogenetic group and epidemiological relationships of EPEC, ETEC and EHEC pathotypes isolated from patients with diarrhea and farm animals in south east region of Iran. A total of 671 diarrheagenic E. coli (DEC) were collected from stool samples of 395 patients with diarrhea and 276 farm cattles and goats. Presence of EPEC, ETEC and EHEC were identified using multiplex-PCR employing primers targeted the shiga toxin (stx), intimin (eae), bundle forming pili (bfp), and enterotoxins (lt and st) genes. The highest proportion of the patients (64%) were children under age 1–15 year (p ≤ 0.05). Among the isolates, atypical EPEC was detected in 26 patients and 14 animal stool samples, while typical EPEC was found in 2 cattles. ETEC isolates were detected in stools of 13 patients and 4 EHEC was identified in 3 goats and one cattle. The isolates were checked for susceptibility to 14 antibiotics. 50% (n = 13) of EPEC and 61.5% (n =8) of ETEC showed multi-drug resistance (MDR) profiles and one EPEC was found to be extensive drug resistant (XDR). In contrast, EHEC isolates were susceptible to the majority of antimicrobial agents. The MDR isolates were positive for bla_TEM and bla_CTX-M ESBL genes and carried class 1 integrons. Further study on the biofilm formation indicated that, 3 out of 4 EHEC isolates showed strong biofilm, while other pathotypes had either moderate, weak or no biofilm activity. Majority of EPEC isolates were belonged to phylogenetic group B1, all except one ETEC were classified as phylogenetic group A and two EHEC were belonged to phylogroup D, respectively. A multilocus variable tandem repeat analysis (MLVA) exhibited 22 distinct patterns. In conclusion, MLVA data showed high clonal diversity. Presence of EHEC in animal origins pose public health concern in this region.     

蒙古黄芪根腐病病原鉴定及防治药剂室内筛选

为鉴定青海省民和地区蒙古黄芪(Astragalus membranaceusvar.mongholicus)根腐病病原菌及筛选防治该病害的有效杀菌剂,本研究基于形态学特征、rDNA-ITS,TEF-1α和RPB2序列分析对蒙古黄芪根腐病病原进行鉴定,并利用菌丝生长速率法测定8种杀菌剂对病原菌的抑制作用。结果发现,引起黄芪根腐病的病原菌为腐皮镰刀菌(Fusarium soani)、逗号镰刀菌(F.virguliforme)、木贼镰刀菌(F.equiseti)和锐顶镰刀菌(F.acuminatum)。室内药剂试验表明硅唑·咪鲜胺对4种镰刀菌的抑制作用最好,EC₅₀在0.195~0.588 mg·L^-1之间;多菌灵对腐皮镰刀菌、木贼镰刀菌和锐顶镰刀菌的抑制作用较强,EC₅₀在0.113~0.869 mg·L^-1之间;咯菌腈对逗号镰刀菌、木贼镰刀菌和锐顶镰刀菌的抑制作用较强,EC₅₀在0.153~0.390 mg·L^-1之间;甲基硫菌灵、噁霉灵、溴菌腈和石硫合剂对4种镰刀菌的抑制作用较差,EC₅₀在1.018~4.360 mg·L^-1之间。试验结果为生产上合理选用杀菌剂防治黄芪根腐病提供了科学依据。     

一种从藤粉蝶分离的微孢子虫的生物学特性和分子系统发育研究

从广州市萝岗桑园周边捕捉到的野外昆虫藤粉蝶(Delias pasithoe)成虫体内分离到1株微孢子虫,经人工接种可在家蚕体内全身性寄生。用透射电子显微镜观察到该微孢子虫为双核,极丝11～12圈,极丝同型,基本符合Nosema属微孢子虫超微结构特征。该微孢子虫在其生活史发育过程中以偶倍数核为主,以二分裂方式增殖,形成双核孢子,发育周期为108 h。基于SSU rRNA和rRNA ITS序列的相似性比较和系统进化分析进一步证实该微孢子虫与Nosema属亲缘关系最近,应被分类为Nosema属‘true’Nosema亚群。     

6个新疆矮化核桃品种的指纹图谱构建及亲缘关系分析

以8个核桃品种叶片为材料,利用ISSR技术,从100条ISSR引物中筛选出2条对8份DNA进行PCR扩增,共扩增位点34个,其中多态位点24个(比例达70.58%),构建出8个核桃品种的数字化指纹图谱。以NTSYSpc2.1软件计算品种指纹图谱的相似系数(GS)并进行(UPGMA)聚类分析,样品间的GS在0.54～0.94之间,在GS=0.64处将8个核桃品种分成2组:第一组母本为‘扎343’(1号),矮化品种有‘新温724’(4号)和‘新温908’(5号);第二组母本为‘新早丰’(2号),矮化品种有‘新温609’(3号)、‘新温915’(6号)、‘新温916’(7号)和‘新温917’(8号),与田间表现基本一致。     

杨树溃疡病病原——葡萄座腔菌科真菌间关系的系统学初探

利用ITS rDNA序列对我国杨树溃疡病的葡萄座腔菌科病原真菌进行系统发育学分析,共得到4种葡萄座腔菌科病原真菌:Botryosphaeria dothidea,Neofusicoccum parvum,Diplodia seriata和D.mutila。它们与相应的无性型或有性型一一对应。其中,D.seriata在我国是第一次报道为杨树的病原。研究了试验菌株与国内外已报道菌株间的系统发育学关系,分析了它们与环境间的联系。结果表明B.dothidea具有高度的种内遗传多样性。这些结果对葡萄座腔菌科真菌在全球的分布作了必要的补充,也为杨树溃疡病的研究和防治提供进一步的理论基础。