盐藻DNA对水稻幼苗在低盐中生长的影响

[目的]分析盐藻DNA对水稻幼苗在低盐中生长的影响。[方法]将刚萌发的水稻幼苗根系浸入5 mg/L盐藻DNA溶液中培养2d,以蒸馏水培养为对照,处理后的水稻幼苗栽于含有氯化钠3 g/L的MS培养液中培养,分析盐藻DNA对水稻幼苗在低盐中生长的影响。[结果]水稻幼苗用5 mg/L盐藻DNA处理后,在含氯化钠3 g/L的MS培养液中培养15 d,其平均株重为150.7 mg,比对照增加了10.3%;平均株高为15.28 cm,比对照增加了6.1%;平均存活率为45.0%,比对照增加了125.0%;平均根数为7.15条,比对照减少了2.8%;平均根长为3.83 cm,比对照缩短了8.4%。说明盐藻DNA提高了水稻幼苗的耐盐适应性。[结论]盐藻DNA可提高水稻的耐盐适应性。     

一种用于桑RAD-Seq高通量测序的基因组DNA提取方法

[目的]筛选一种能够得到高质量桑树基因组DNA的方法。[方法]采用天根试剂盒法(Plant Genomic DNA Kit离心柱型),并稍加改进,从11个桑种的新鲜嫩叶中提取DNA。[结果]该方法简单快速,不需要复杂仪器,且能有效地除去桑叶中的多糖、多酚、蛋白质等杂质,得到的DNA能够满足RAD-Seq高通量的测序要求。[结论]建立了一种用于桑RAD-Seq高通量测序的基因组DNA提取方法。     

中药植物川牛膝基因组从头测序及分析

川牛膝是我国重要大宗药材,但近年来品质退化严重,品种真伪混杂。为深入研究川牛膝有效成分积累的分子基础,采用第二代测序技术利用Illumina Hi-seq 2000测序平台对川牛膝进行全基因组测序,使用AByss进行初步组装,得到一个包含大量基因组序列信息的数据集,并对其进行重复序列及编码序列注释。在该测序结果基础上,初步预测得到川牛膝体内甾体合成途径,并对鉴定川牛膝真伪的SCAR分子标记进行了初步定位,为研究川牛膝主要有效成分杯苋甾酮的合成途径,改良川牛膝品质提供了基础。     

DNA折纸术的研究进展

为了帮助广大研究者在短时间内了解DNA折纸术的最新研究现状,掌握DNA折纸术相关软件的应用,本论文将DNA折纸术的相关内容进行了归纳。首先介绍了DNA折纸术的起源与基本原理;然后综述了近几年国内外的研究进展与取得的成果;接下来对DNA折纸术相关软件-Tiamat和Cadnano的应用和功能进行了简单的介绍,并对不同的软件进行了比较;最后对DNA折纸术的应用前景进行了展望。DNA折纸术虽然取得了一定的研究进展,但其未来的发展和应用前景将更广阔。     

鸡传染性支气管炎病毒SD060311株和GD080122(肾型)株的全基因组序列分析

通过对鸡传染性支气管炎病毒(IBV)分离株SD060311和GD080122(肾型)的基因组进行RT-PCR扩增和基因序列测定,结果表明其基因组大小分别为27 788 nt和27 407 nt,与已报道的IBV全基因组序列大小不完全一致,但基因组编码基因的顺序与已报道的IBV一致,均为5′cap-Replicase-S-3a-3b-3c-M-5a-5b-N-poly (A)3 ′.与已报道的IBV毒株和火鸡冠状病毒进行比较,绘制系统进化树,结果显示,SD060311和GD080122(肾型)分离株自成一支,与中国分离株BJ株和A2株的亲缘关系最近.本研究不仅丰富了冠状病毒的生物信息数据库,且为进一步研究IBV致病机理和变异机制等奠定了基础.     

Comparison of DNA probe test and cultivation methods for detection of Mycobacterium avium subsp paratuberculosis in caprine and ovine faeces

OBJECTIVE: To compare a DNA probe test with two cultivation methods for the detection of Mycobacterium avium subsp paratuberculosis in goat and sheep faeces. DESIGN: Comparison of the results of the three methods with histological examination as the reference standard. PROCEDURE: Faecal specimens were obtained from goats and sheep originating from flocks known to be affected with paratuberculosis and tested for Mycobacterium avium subsp paratuberculosis with a DNA probe test and two cultivation methods (old conventional culture and new double incubation method in Herrold's and Lowenstein-Jensen medium). RESULTS: In goats, the sensitivity of the various tests were for the DNA probe test 17.2%, for the double incubation culture method 25.4% and for the old conventional culture method 22.8% using the histopathological results as reference. In sheep the sensitivity of the various tests were for the DNA probe test 13.2%, for the double incubation culture method 8.8% and for the old conventional culture method 5.9% using the histopathological results as reference. The specificity of the above tests was 100% in goats and sheep and the specificity of the double incubation culture method in goats was 91%. CONCLUSIONS: The DNA probe test is a rapid and specific test that could be used in a control program if the sensitivity of the test were improved and possibly in combination with another test.     

Demonstration of Heterogeneous Genotypes of Taylorella equigenitalis Isolated from Horses in Six European Countries by Pulsed-Field Gel Electrophoresis

Forty-six isolates of Taylorella equigenitalis were analysed by pulsed-field gel electrophoresis (PFGE) after separate digestion of the genomic DNA with ApaI and with NotI. The isolates had been obtained from horses in six European countries and were classified into 18 genotypes. In Belgium, 2 genotypes were detected in 2 isolates, in England 9 among 15, in Finland 2 in 2, in France 2 among 10, in Sweden 3 among 5, and in Switzerland 3 among 12. Two English isolates and 4 French isolates gave identical PFGE profiles to those of Kentucky 188 from the United States. A common genotype was found in 5 isolates from Belgium and England and also in 10 isolates from France and Switzerland. The analysis of genomic DNA from 12 isolates of T. equigenitalis obtained from male horses in France, Sweden and Switzerland gave no evidence of a sex-related difference in the genomic DNA. Genomic DNA from 11 streptomycin (STM)-susceptible isolates obtained in Sweden and Switzerland were classified into four genotypes by PFGE. Each of the six genotypes determined among the 17 isolates from these two countries had single phenotypes for resistance or susceptibility to STM.     

Application of a putative fatty-acid binding protein to discriminate serologically the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from other Meloidogyne species

Two major proteins, Mcf-A67 and Mcf-B66, were identified by mini two-dimensional polyacrylamide gel electrophoresis in order to distinguish the two European quarantine root-knot nematodes, Meloidogyne chitwoodi and M. fallax, from eight other species. These quarantine proteinic markers have been microsequenced after enzymatic digestion. The internal amino acid sequences exhibit similarities to members of a family of low molecular weight intracellular lipid-binding proteins. Moreover, to explore a simple, rapid, and inexpensive way to identify the two quarantine nematodes, dot blot hybridizations were performed using an antiserum (A67) produced from the longest amino-acid sequence of the protein Mcf-A67. Although several proteins stained on the M. chitwoodi and M. fallax western blot membranes, the two nematodes were easily distinguished from other root-knot nematodes, on dot blot assays with soluble proteins extracted from a single female. Because of its specificity and sensitivity, the use of the A67 antiserum to improve the diagnosis of the two European quarantine root-knot nematodes is discussed.     

Genetic relationships between the dominant genotypes of Phytophthora infestans in Hokkaido, Japan

The genetic characteristics of the dominant genotypes of Phytophthora infestans in Japan (US-1, JP-1, Japanese A1-A, A1-B) were compared. Differences were evident in the peptidase genotype, amplified fragment length polymorphism, and RG57 DNA fingerprints. Almost all of the fingerprint bands for the Japanese genotype A1-B were also present in JP-1 and Japanese A1-A, and few bands were unique to Japanese A1-B. These results suggest that the Japanese A1-B genotype was generated from sexual reproduction involving Japanese A1-A and JP-1 or related genotypes.     

Random amplified polymorphic DNA markers reveal low genetic variation and a single dominant genotype in Eichhornia crassipes populations throughout China

Genetic variation within and between 34 populations of Eichhornia crassipes (water hyacinth) in China was surveyed using random amplified polymorphic DNA (RAPD) markers. A total of 1009 individuals were analysed, for which 12 RAPD primers amplified 69 reproducible bands, with 22 (32%) being polymorphic. The percentage of polymorphic loci (p) within a population ranged from 4.4% to 17.4%, and the mean Nei's gene diversity (H_e) was 0.046 ± 0.0145, indicating a low genetic diversity of E. crassipes in China. Each population contained at least four RAPD phenotypes (genotypes), and the same particular genotype was invariably dominant in all the populations sampled. The mean proportion of distinguishable genotypes was 0.29. Analysis of molecular variance revealed a large proportion of genetic variation (83.9%) residing within populations and a slightly larger proportion (88.1%) within localities, indicating a low genetic differentiation of E. crassipes populations, both locally and regionally. Human-mediated dispersal, vigorous clonal growth, and a generally low level of sexual reproduction were thought to be responsible for such a pattern of genetic structure.