朱顶红幼嫩花梗胚性愈伤组织诱导和高效植株再生

为建立高效的朱顶红植株再生和种苗繁育技术体系，以幼嫩花梗为外植体开展胚性愈伤组织诱导和植株再生研究。对2,4-D和噻苯隆（TDZ）浓度、外植体发育时期及大小进行了筛选，结果表明：将切片厚度为1 mm的幼嫩花梗外植体置于添加0.5 mg ? L-1 TDZ 和2.0 mg ? L-1 2,4-D的MS固体培养基上培养8周，胚性愈伤组织诱导率最高，达85.3%。将胚性愈伤组织转移至相同的培养基上，每月继代1次，平均每月增殖10.6倍。在不含任何生长调节剂的MS培养基上，胚性愈伤组织的植株再生率达98.0%，平均每块愈伤组织可再生出12.3个小植株。经过36次（3年）继代培养后，胚性愈伤组织的增殖和植株再生效率没有显著变化。幼苗移栽至温室驯化，成活率达97.5%。再生植株移栽到田间，没有发现明显的表型变异。对随机选择的再生植株和母株进行简单序列重复区间（ISSR）扩增分析，证实再生植株没有发生DNA水平变异。     

Genetic diversity and aggressiveness of Calonectria pteridis in Eucalyptus spp.

Calonectria leaf blight, caused by Calonectria pteridis, is currently one of the main foliar diseases in eucalypt plantations in Brazil. In warm and high rainfall regions, the disease can be a limiting factor for eucalypt production when planting susceptible genotypes. The most effective method for controlling this disease in the field is the use of resistant genotypes, which requires knowledge of the genetic variability and aggressiveness of the pathogen population for effective deployment of plant resistance. This work evaluated the genetic diversity and aggressiveness of C. pteridis populations obtained from infected eucalypt plants in Monte Dourado (Pará state) and Imperatriz (Maranhão state), Brazil. To study the genetic diversity, 16 ISSR primers were tested, five of which amplified polymorphic, reproducible and informative bands. Thirty-one closely related genotypes were identified from 84 isolates studied, indicating that the population has a low genetic diversity. The aggressiveness of seven isolates, selected according to geographic origin and their clustering in the ISSR-based dendogram, was determined by inoculation of a hybrid Eucalyptus grandis × E. urophylla clone under controlled conditions. Disease severity was assessed by both measuring the percentage of plant defoliation and assigning a score according to a diagrammatic scale of symptoms. A high correlation between the two evaluation methods was observed, which revealed significant differences in aggressiveness among the isolates. The diagrammatic scale is recommended for disease evaluation because results are obtained much faster, before the occurrence of severe defoliation. No correlation between clustering in the ISSR-based phylogenetic analysis and aggressiveness was observed.     

香樟ISSR-PCR反应体系的正交优化

应用L9(34)正交表建立了适合于樟树ISSR-PCR的优化反应体系,即20μL ISSR反应体系中含有1×PCR buffer、dNTP0.25 mmol/L、引物0.3μmol/L、Mg2 2.0 mmol/L、Taq酶1 U和模板DNA50 ng.适宜的扩增条件为:94℃预变性4 min;94℃变性40 s,58℃退火1 min,72℃延伸2 min,42个循环;72℃延伸10 min;4℃保存.     

云南主要核桃品种的ISSR分子鉴别

以核桃当年萌发的新叶为材料,在建立了良好的反应体系基础上,筛选出8条引物对云南省11个主要栽培和推广的核桃品种进行了ISSR分子标记研究。结果表明:8个ISSR引物共检测出位点102个,其中多态性位点62个,占60.78%,参试样品具有较高的多态比率;用其中的两条引物建立了11个品种的指纹图谱,并可用之鉴别参试品种;POPGENE 32软件的计算结果有效等位基因数、基因的多样性和Shannon信息指数分别为1.370 7,0.214 3和0.321 6,供试的云南核桃品种在分子水平上存在比较丰富的遗传变异。     

Genetic diversity of sea buckthorn (Hippophae rhamnoides) populations in northeastern and northwestern China as revealed by ISSR markers

As a N₂-fixing tree species, sea buckthorn (Hippophae rhamnoides) is well adapted to arid regions and is utilized for multiple purposes in China. Current knowledge of genetic variability of H. rhamnoides is limited in terms of rangewide distributions. Eleven natural populations of sea buckthorn in northeastern and northwestern China were analyzed to detect genetic variation among and within populations, by use of ISSR (inter-simple sequence repeats) markers. Using eight primers, 207 polymorphic loci were observed, ranging in size from 250 to 2500 bp. The coefficient of gene differentiation (Gst = 0.0679) showed that the total molecular variance of 11 populations was mainly existed within populations. The genetic variation within and among the 11 populations was 93.21 and 6.79%, respectively. No significant correlation between genetic and geographic distances of the populations was found using ISSR markers. Our study provides a population-level genetic profile for further investigation and conservation of genetic diversity of sea buckthorn.     

广东省木棉种质资源ISSR遗传多样性分析

利用ISSR分子标记技术对144份木棉（Bombax ceiba）种质资源遗传多样性进行分析。结果表明: 木棉种质资源具有较高的遗传多样性,采用筛选的26个ISSR引物共扩增出179个位点,多态性位点161个,多态性位点百分率为899%,有效等位基因数为16,Nei基因多样性指数为0350 5, Shannon信息多样性指数为0523 3,相似性系数为0238 6～0910 6。UPGMA聚类分析得出,木棉种质之间的亲缘关系与来源没有明显的相关性,对同一产地来说,绝大部分种质聚在一起,亲缘关系较近,同时不同产地之间多数木棉种质亲缘关系也较近。     

天然红松种群遗传多样性和遗传分化的研究

ISSR是一种特异性较强、稳定性较高的分子标记方法。本研究采用ISSR-PCR技术,检测了红松(PinuskoraiensisSieb.etZucc)在伊春市汤旺河高峰林场、长白山二道白河、黑河胜山林场和俄罗斯海参崴市郊的四个种群的遗传多样性和遗传分化。15个引物的扩增结果表明:红松群体的多态位点比率是60.70%,平均每个引物3.6个多态位点,多样性水平在松科植物中是较高的;红松分布中心区的遗传多样性要高于边界区;和大多数木本植物一致,红松群体的基因多样性主要来自种群内部,占总基因多样性的73%;红松四个种群的遗传距离和地理距离之间无正相关性。根据研究结果推测,天然红松分布区逐渐缩小的原因不是由于遗传多样性水平过低引起的,而是由于人类的破坏作用,再加上火灾和风倒等因素造成的。     

凤凰单丛茶树资源遗传多样性的ISSR分析

采用ISSR分子标记技术对48份凤凰单丛茶品种(系)进行了遗传多样性分析。选用10个多态性高、分辨力强的ISSR引物分别对供试材料基因组DNA进行扩增,共获得121个位点,其中多态位点带98个,多态位点比率(PPB)为81.0%。品种(系)之间的遗传相似系数变化范围在0.590 9~0.967 4之间。UPGMA聚类分析结果表明,在GS值为0.792的水平上供试材料可划分为7大类,其亲缘关系远近与地理来源关系不大且与香味类型没有必然的联系。     

赣江中游四大家鱼遗传多样性的ISSR分析

利用ISSR分子标记技术对赣江中游四大家鱼的遗传多样性进行分析,青鱼（Mylopharyngodon piceus）、草鱼（Ctenopharyngodon idellus）、鲢（Hypophthalmichthys molitrix）、鳙（Aristichthys nobilis）分别扩增出47、74、55、59个位...     

新疆8个棉花品种（系）的指纹图谱分析

利用ISSR对8个新疆品种（系）进行指纹图谱构建。通过基因组多态性分析，从60条引物中筛选到11条扩增效果好的引物，用其中2条引物UBC809，UBC841建立8个品种的指纹图谱，统计品种鉴定的置信概率达到1／8388608。结果显示，ISSR分子标记技术对棉花品种的指纹图谱构建具有高效性和准确性。