Orobanche species and population discrimination using intersimple sequence repeat (ISSR)

Summary Orobanche species are commonly identified using morphological characteristics. In many cases, the distinction of closely related species is difficult, and a molecular tool is more suitable to differentiate them. In this study, genomic polymorphism between morphologically distinct species was investigated through amplification by polymerase chain reaction (PCR) of intersimple sequence repeat (ISSR) regions. Five primers were used to study genetic variation in the morphologically distinct species O. hederae and O. amethystea, as well as the closely related species O. cernua and O. cumana. For the first two species, all the primers detected genetic polymorphism. Anchored primers allowed the identification of more specific molecular markers than non‐anchored tri‐ and tetranucleotide primers. Genetic polymorphism was investigated among three O. hederae populations using the two types of primer. One non‐anchored and two anchored primers detected intraspecific variation, which was not correlated with the geographical location of those populations. The primer (GATA)₄ detected polymorphism between five specimens each of O. cernua and O. cumana species collected from different countries, permitting these two closely related species to be clearly differentiated. This study demonstrated that ISSR markers can be highly reliable for precise identification of Orobanche species.     

Cardiac troponin I in canine patients with lymphoma and osteosarcoma receiving doxorubicin: comparison with clinical heart disease in a retrospective analysis

The cumulative cardiotoxicity that occurs as a result of doxorubicin chemotherapy is irreversible and can affect both quality and quantity of life for the cancer patient. Cardiac troponin I (cTnI) is a sensitive and specific marker of cardiomyocyte death. The purpose of this retrospective study was to evaluate serum concentrations of cTnI in dogs with lymphoma or osteosarcoma given doxorubicin chemotherapy, and with known cardiac outcome, based on a minimum assessment by physical examination and thoracic radiography. Serum samples were also available for cTnI measurement from seven healthy dogs given intracoronary doxorubicin. Serial serum samples obtained before, during and after doxorubicin chemotherapy showed increased cTnI concentrations in some clinical patients following chemotherapy (P = 0.0083 compared to baseline), but this did not correlate with clinical signs of cardiomyopathy. In dogs that subsequently developed cardiomyopathy however, serum cTnI concentrations were elevated before clinical signs became evident (confirmed with echocardiography).     

玉米抗粗缩病遗传及分子标记研究

Maize rough dwarf disease caused by Rice black-streaked dwarf virus (RBSDV) is transmitted by planthopper in China. Identification and development of resistant hybrids are complicated because of the inconsistencies in viral disease pressure every year. Marker-assisted selection can provide means for main-taining virus resistance alleles even in the absence of disease. In this paper a F₂ segregation population was constructed to identity the molecular markers linked to the resistance gene using a cross between a resistant and a susceptible parents (Qi319×Ye107). Fifteen-day-old seedlings of F₂ population were exposed to small brown planthoppers carrying RBSDV for 3 days in specific inoculation chamber. The inoculated plants were transplanted to screenhouse after removing the insects completely. In plant maturity stage the disease resistance of all the individuals were visually assessed. The results showed that 17, 8, 11, 51 and 122 plants were scaled from 0-4 respectively, in which 0 means no symptoms and 4 represents highly susceptible. Chi-square test demonstrated that the segregation ratio of phenotype was 1∶15 (resistant: susceptible) or 1∶6∶9 (resistant∶moderate∶susceptible) in the F₂ population, indicating RBSDV resistance of maize was controlled by two recessive genes. The F₂ individuals DNA were extracted and 261 SSR (simple sequence repeat) primers derived from maize genome ten chromosomes were selected from maize GDB database to construct genetic linkage map. The linkage map consisted of 71 polymorphic SSR markers, spanning a genetic distance of 996.6 cM with an average interval of 14.0 cM between adjacent markers. The resistant and susceptible gene pools were set up for BSA (bulked segregant analysis) and 6 polymorphism markers were obtained with BSA-SSR method between the two pools. The F₂individuals were further analyzed with 6 polymorphism markers. Chi-square test showed that phi 051, umc1407 and umc1432, mapped on chromosome 7 and 10, exhibited segregation distortion significantly and very significantly in susceptible individuals. These three SSR markers were identified as potential markers linked to the resistant loci.     

基于表型和基因型的藜麦种质资源遗传多样性分析

为了解和掌握青海省引进和收集藜麦种质资源的遗传背景和多样性,本研究选择11个表型性状和221对SSR引物对114份藜麦种质资源进行遗传多样性分析。结果表明,114份藜麦种质资源的11个表型性状差异极显著（P＜0.01）,变异系数范围为4.57%~87.79%,主穗直径的变异最大（87.79%）,籽粒直径的变异最小（4.57%）;遗传多样性指数范围为0.47~2.29,单株产量的遗传多样性指数最大（2.29）,茎秆长最小（0.47）。46对多态性较好的SSR引物扩增出165条多态性条带,每对引物的平均等位变异数为3.59,观测等位基因（Na）平均值为1.65,有效等位基因（Ne）平均值为1.50,Shannon’s信息指数（I）和Nei’s基因多样性指数（He）平均值分别为0.39和0.27,平均多态性比率为64.35%。通过聚类分析发现,114份藜麦种质基于表型性状和SSR分子标记的聚类结果存在一定的相似和差异,来源地相同的种质被分到了不同的类群,来源地不同的种质被分到了同一类或单独聚成一类,说明参试藜麦种质资源的表型性状和分子遗传信息多样性较丰富。     

浙江沿海三疣梭子蟹的养殖模式

为开发三疣梭子蟹低盐耐受性相关甲基化标记,采用甲基化敏感性高分辨率熔解曲线法(methylation sensitive high resolution melting curve, MS-HRM)进行了三疣梭子蟹低盐耐受性相关甲基化位点的筛选和应用。结果显示,从三疣梭子蟹转录组数据库中共开发了8个甲基化标记,其中6个标记与低盐度耐受性显著相关,在低盐度条件下表现出显著的去甲基化或甲基化,这些标记分别位于V-ATP酶(V-type proton ATPase)、CLC型氯离子通道、琥珀酸脱氢酶、NAD(P)转氢酶(NAD(P)、磷酸乙醇酸性磷酸酶、NADH还原酶基因,为三疣梭子蟹耐低盐新品种的分子标记辅助选育提供候选工具。研究表明,“宁象1号”耐低盐能力显著高于野生群体,且选育群体中位点Pt-M1、Pt-M2的去甲基化率达到100%,可能有利于低盐条件下保持体内Na⁺和Cl⁻的平衡。     

思茅松EST-SSR标记在几种松属植物中的通用性分析

为验证思茅松EST-SSR标记在松属植物中的通用性,利用已获得的18对EST-SSR标记对其近缘种进行扩增。结果表明：思茅松EST-SSR标记对马尾松、高山松、油松和云南松的通用率为88.89%,多态性百分比率分别为81.25%、81.25%、75.00%和93.75%。因此,基于思茅松转录组序列SSR标记开发是可行的,思茅松EST-SSR标记对近缘种具有较高的通用性和多态性,这些引物可用于分析思茅松及其近缘种间的遗传关系。     

不同山茱萸种质资源形态和ISSR遗传多样性研究

利用形态特征和ISSR标记对35份山茱萸种质资源进行遗传多样性分析,利用NTSYSpc-2．11F软件分析遗传相似系数,UPGMA方法聚类,构建亲缘关系系统图。结果表明,35份山茱萸在9个性状上差异明显;分子标记中9条引物共得到条扩增条带179条,其中有164条呈现多态性,占91．6％,遗传相似系数变化范围O．575-0．877。对其进行数据化处理后聚类,35份山茱萸材料可划分为2个类群6小组,材料的聚类结果与地理分布、形态特征有一定的相关性。     

DNA分子标记在甘薯遗传育种中的应用

DNA分子标记是继形态标记、细胞标记和生化标记之后发展起来的一种新的较为理想的遗传标记,在甘薯育种方面发挥了重要作用。主要概述了分子标记在甘薯的起源、进化与分类、图谱构建、遗传多样性分析、品种鉴定、基因定位与辅助育种等方面的研究进展,并对甘薯的分子育种提出了展望。     

我国北方地区花生品种的遗传多样性分析

利用SSR分子标记对我国北方地区育种品种以及区试品种共计68个栽培花生品种进行遗传多样性分析,结果表明,104对SSR引物中有15对在品种间存在多态,多态率为14.4%,平均每个标记产生3.8个位点。15对引物的多态信息量在0.397~0.667之间。山东地区花生品种的遗传多样性要高于我国北方其他地区供试品种。小粒组品种的遗传多样性要略高于大粒组品种。聚类分析表明我国北方68个育成和区试品种的遗传相似系数在0.64~0.91之间,分成两个大类,第一个大类可分成四小类,其中第三小类又可以分成六个亚类。这些结果表明我国北方地区现有育成和区试品种的遗传多样性较低,为拓宽花生育种的遗传基础,应从中发掘出有利用价值的品种。