亚硫酸钠和亚硫酸氢钠混合液对2种云杉某些生理指标影响的比较

八年生嫩江云杉和红皮云杉苗木,经喷施20、50、100mmol·L-1Na2SO3和NaHSO3(摩尔浓度为3:1)的混合液后第3、11、20天测定多个生理指标的结果显示,喷后第3天,2种云杉的净光合速率(Pn),光系统II(PSII)的最大光能转换效率(Fv/Fm),光化学量子效率(ΦPSII),叶绿素a(Chla)、叶绿素b(Chlb)和类胡萝卜素含量均下降;相对电导率上升;苗木受到胁迫的程度随着Na2SO3和NaHSO3混合液浓度的提高而增大。喷后第8~20天各种生理指标开始恢复。气孔导度(Gs)和胞间CO2浓度(Ci)变化不显著。红皮云杉对胁迫的敏感度高于嫩江云杉,但其在重度胁迫后的修复能力强于嫩江云杉。     

木霉肽类代谢产物对豇豆土著根瘤菌遗传性状的影响

利用RAPD与ISSR分子标记检测手段,分析了哈茨木霉T2-16肽类代谢产物处理豇豆土著根瘤菌,对其遗传性状的影响,同时,比较了RAPD和ISSR两种不同分子标记在检测根瘤菌种间的遗传相似性以及遗传变异性的分辨力.实验中,从100条引物中筛选到具有多态性的ISSR引物5条,从80条引物中筛选到具有多态性的RAPD引物6条,用5条ISSR引物扩增出54条带,多态性条带比率为75.93 %;6条RAPD引物扩增出61条带,多态性条带比率为68.85 %.两种分子标记均能揭示出处理前后根瘤菌间的遗传差异,但ISSR标记比RAPD标记可检测到更大的遗传变异.根据两种标记的结果,对供试的根瘤菌进行聚类分析,结果表明,土著根瘤菌经木霉肽类代谢产物处理后,与出发菌株相比,表现出一定程度的遗传分化和遗传差异性.     

叶子花种质资源遗传多样性及亲缘关系的RAPD和ISSR分析

利用RAPD和ISSR标记对48份叶子花种质进行遗传多样性及亲缘关系分析。结果显示:(1)筛选出具有多态性的RAPD引物7条、ISSR引物11条,RAPD引物共扩增97条多态性条带,ISSR引物共扩增140条多态性带,多态性百分率均达100%。(2)根据两种标记的扩增结果,用UPGMA法对48份叶子花种质的聚类分析显示,48份供试材料间具有较丰富的遗传多样性,其品种间遗传相似系数RAPD为0.318 8～0.955 6,ISSR为0.349 5～0.900 0;两种分子标记均能清楚地将48份种质材料区分开来,对种质类群的划分结果基本一致,仅有些许差异。(3)聚类分析结果显示,48份种质可分为两大种系:1)B.glabra种系,其品种大多以其种内来源为主;2)涵盖了B.spectabilis、B.peruviana、B.×buttiana和B.×spectoglabra等4个种的种系,种质构成较为复杂。(4)两种标记聚类结果呈显著相关关系,相关系数为0.752 3。研究表明,对一些形态上无法细分的叶子花种质(类群),RAPD和ISSR分子标记是可靠的鉴别方法。     

利用RAPD和ISSR标记分析烤烟品种间遗传关系

利用RAPD和ISSR标记对22份烤烟(Nicotiana tabacumL.)品种进行了遗传关系研究。在RAPD分析中筛选到13个引物,共扩增出167条带,其中多态性带50条,多态性比率为29.9%;在ISSR分析中筛选出7个引物,共扩增出96条带,其中多态性带44条,多态性比率为45.8%。两种标记相结合估算出的品种间遗传相似系数在0.881~0.979之间,平均为0.933。单独基于RAPD标记和ISSR标记的聚类结果有一定差异;两种标记结合起来的聚类分析结果与系谱信息吻合程度更高。定向选择可能对烤烟品种间遗传关系有较大影响;国外引进品种与国内育成品种并未完全分开,表明分子水平的遗传关系和地理来源间缺乏必然联系。     

水分胁迫对云杉光合特性的影响

比较研究了3年生红皮云杉和嫩江云杉苗木在不同水分条件下的光合特征,并探讨其对水分胁迫的适应机制。研究结果表明,不同光强下两种云杉的最大光合速率（P_max）随土壤含水量的降低而逐渐下降,且嫩江云杉的降幅较小;表观量子效率（AQY）、蒸腾速率（Tr）、气孔导度（G_s）也都呈下降的趋势。暗呼吸速率（R_d）和水分利用率（WUE）在逐渐干旱的过程中先上升后下降,而胞间CO₂浓度先下降,后随上升。这表明在水分胁迫较为严重时,光合速率的降低主要是非气孔限制造成的。比较上述指标的变化情况可以看出,嫩江云杉对干旱的适应性要强于红皮云杉。     

基于RAPD、ISSR和AFLP对西瓜枯萎病菌遗传多样性的评价

利用RAPD、ISSR和AFLP分子标记技术对50个西瓜枯萎病菌株进行了分析。结果表明,21个RAPD引物、21个ISSR引物和21对AFLP引物分别对供试菌株扩增出113、134和389条带,三种分子标记的遗传相似系数比较一致,均可揭示西瓜枯萎病菌的遗传变异特点。三种分子标记产生的聚类分析结果存在一定差异,其中RAPD类群与生理小种和地理来源之间均不存在明显关系;而AFLP和ISSR类群与生理小种之间存在一定相关性,与菌株的地理来源关系不明显.     

家蚕胚胎细胞系的DNA指纹图谱分析

在建立可靠的家蚕细胞系基因组DNA制备和PCR扩增技术体系的基础上,筛选具有稳定多态性位点的RAPD和ISSR引物,建立家蚕细胞系基因组DNA的ISSR和RAPD分子标记技术体系,检测家蚕细胞系的DNA分子标记多态性,构建细胞系的DNA指纹图谱。筛选出了26个ISSR引物和43个RAPD引物,通过PCR扩增在家蚕胚胎细胞系和传代昆虫细胞系等9个样品中分别获得了797条和1205条多态性条带,多态性达到89．9%和76．6%,不同细胞系的DNA多态性有较大差异,三个家蚕胚胎细胞系具有各自特有的DNA标记。测定了9个样品间的Nei's相似系数和遗传距离,构建了系统发育树,结果表明本实验室建立的3个家蚕胚胎细胞系和家蚕“夏芳×秋白”聚为一簇,亲缘关系较近,而来自不同物种的五个传代昆虫细胞系聚为一簇,它们之间的遗传距离比3个家蚕胚胎细胞系之间的遗传距离更小。     

基于RAPD、ISSR和AFLP对西瓜枯萎病菌遗传多样性的评价

利用RAPD、ISSR和AFLP分子标记技术对50个西瓜枯萎病菌株进行了分析。结果表明,21个RAPD引物、21个ISSR引物和21对AFLP引物分别对供试菌株扩增出113、134和389条带,三种分子标记的遗传相似系数比较一致,均可揭示西瓜枯萎病菌的遗传变异特点。三种分子标记产生的聚类分析结果存在一定差异,其中RAPD类群与生理小种和地理来源之间均不存在明显关系;而AFLP和ISSR类群与生理小种之间存在一定相关性,与菌株的地理来源关系不明显。     

基于RAPD、ISSR和AFLP对西瓜枯萎病菌遗传多样性的评价

利用RAPD、ISSR和AFLP分子标记技术对50个西瓜枯萎病菌株进行了分析。结果表明,21个RAPD引物、21个ISSR引物和21对AFLP引物分别对供试菌株扩增出113、134和389条带,三种分子标记的遗传相似系数比较一致,均可揭示西瓜枯萎病菌的遗传变异特点。三种分子标记产生的聚类分析结果存在一定差异,其中RAPD类群与生理小种和地理来源之间均不存在明显关系;而AFLP和ISSR类群与生理小种之间存在一定相关性,与菌株的地理来源关系不明显。     

利用RAPD和ISSR分子标记分析地黄种质遗传多样性

用RAPD与ISSR技术对地黄的8个品种和2个脱毒品系进行了种质遗传多样性分析.分别从80条RAPD引物和44条ISSR引物中筛选出适合地黄种质分析的17条RAPD引物和10条ISSR引物用于RAPD和ISSR分析.17条RAPD引物共扩增出177条带, 多态性位点数为109; 多态性位点比率为61.58%;平均多样性指数(I)为0.3135;每个位点的有效等位基因数(Ne)是1.3641; 10条ISSR引物共扩增出110条带. 多态性位点数为79; 多态性位点比率为71.58%;平均多样性指数(I)为0.3577;每个位点的有效等位基因数(Ne)是1.4037. 基于扩增条带数据库建立了各自的Jaccard遗传相关系数矩阵,构建了相似的分子树状图,将10个供试材料分为2类:一类群含组培85.5、大田85.5、组培9302、大田9302、金状元和金白6个材料;另一类群含北京1号、大红袍、地黄9104和野生地黄4个材料.两种分子标记的分析结果呈极显著正相关(r＝0.649).结果表明,RAPD与ISSR标记适合于地黄种质遗传多样性分析,ISSR标记技术是一种多态性和重复性优于RAPD技术的实用技术.     

水分条件对红松和西伯利亚红松针叶脯氨酸与叶绿素含量的影响

以盆栽3年生红松和西伯利亚红松为材料,设置4种水分条件,土壤含水量分别为：29％～31％（C）, 22%～24%（L）, 15%～17%（M）和渍水组（W）,研究不同水分条件下红松和西伯利亚红松当年生针叶和往年生针叶的脯氨酸含量和叶绿素含量变化。结果表明：1）脯氨酸含量在不同叶龄和不同树种之间存在差异。红松当年生针叶叶绿素含量高于往年生针叶,而西伯利亚红松则相反。总体上西伯利亚红松的脯氨酸含量高于红松;2）2树种在渍水条件下脯氨酸含量大量积累,红松在处理后一个月即出现胁迫反应,早于西伯利亚红松。土壤含水量在 15%～17%时已对2种红松的往年生针叶产生胁迫,但对红松的胁迫程度大于西伯利亚红松;3）叶绿素积累与2种红松的耐水分胁迫能力相关性不大;4）西伯利亚红松的水分适应范围大于红松;2种红松的当年生针叶的耐水分胁迫能力均大于往年生针叶。     

红松天然林种内和种间竞争关系的研究

基于穆棱林业局3块红松天然林标准地调查数据,采用竞争区域动态半径的概念确定了林分中有效竞争木,并利用Hegyi竞争指数对红松天然林的种内、种间竞争强度进行了定量分析。研究结果表明：红松的种内和种间竞争强度分别占总竞争强度的18.0%和82.0%,说明红松天然林种内竞争小,主要竞争来自种间。色木、云杉、冷杉林木个体高大,数量较多,在群落中占据一定优势,对红松的竞争压力较大。红松对象木胸径与竞争指数之间呈幂函数关系,随着红松胸径的增大,受到的竞争强度逐渐减小。当红松胸径小于25 cm时,所受到的竞争压力较大;当胸径大于25 cm时,竞争指数较小,并维持稳定。     

Species-diagnostic and species-specific DNA sequences evenly distributed throughout pine and spruce chromosomes

Genome organization in the family Pinaceae is complex and largely unknown. The main purpose of the present study was to develop and physically map species-diagnostic and species-specific molecular markers in pine and spruce. Five RAPD (random amplified polymorphic DNA) and one ISSR (inter-simple sequence repeat) species-diagnostic or species-specific markers for Picea mariana, Picea rubens, Pinus strobus, or Pinus monticola were identified, cloned, and sequenced. In situ hybridization of these sequences to spruce and pine chromosomes showed the sequences to be present in high copy number and evenly distributed throughout the genome. The analysis of centromeric and telomeric regions revealed the absence of significant clustering of species-diagnostic and species-specific sequences in all the chromosomes of the four species studied. Both RAPD and ISSR markers showed similar patterns.     

RAPD variations among pure and hybrid populations ofPicea mariana, P. rubens, andP. glauca (Pinaceae), and cytogenetic stability ofPicea hybrids: Identification of species-specific RAPD markers

Random amplified polymorphic DNA (RAPD) analysis was used to determine genetic relationships amongP. mariana (black spruce),P. rubens (red spruce), andP. glauca (white spruce) and to assess the degree of polymorphism within populations from different provenances and among spruce hybrids. Eleven arbitrary decamer primers were used to amplify genomic DNAs extracted from embryogenic cultures and seedlings. Species-specific RAPD markers were identified.Picea mariana andP. rubens showed similar RAPD profiles confirming their close genetic relationship. Species-specific RAPD markers were identified and were useful in distinguishing white spruce from black and red spruces. RAPD differentiation between populations within each species was small. The level of polymorphism was much higher in spruce hybrid populations than in the pure species. Cytological analysis ofP. mariana ×P. rubens hybrids showed normal mitotic behaviour at prophase, metaphase, anaphase, and telophase. All the hybrids analyzed from different cross combinations were euploids.     

黑龙江省东部地区红松人工中幼龄林生物量研究

运用非线性联立方程组建模理论建立红松立木相容性生物量模型,然后利用模型计算出人工红松各个样地林木各器官和样地总生物量。以林分年龄、林分平均胸径、林分密度等因素为制约条件,讨论分析林分生物量在林木各器官之间的分配规律,并且探究林分年龄、林木大小和林分密度的变化对林分生物量的影响。结果表明:幼龄红松人工林林分生物量与平均胸径成正相关关系;林分密度对林分生物量影响较大,并且随着密度的增大而增大,且最适合的林分密度范围是1 000~1 400株·ha-1;红松人工幼、中龄林林分生物量各器官分配规律相同,表现为树干树根树枝树叶,地上生物量占林分生物量79%左右;林分地上和地下生物量大概呈3.8∶1的比例。     

Genetic validation and characterization of RAPD markers differentiating black and red spruces: molecular certification of spruce trees and hybrids

 Picea mariana (black spruce) and P. rubens (red spruce) are closely related species which are difficult to differentiate morphologically. RAPD markers differentiating black and red spruces have been previously identified. In the present study, genetic validity of these markers was determined using samples representing range–wide provenances. Their applicability for certifying genetic identity of individual black, red trees and their hybrids from several sympatric and allopatric locations was demonstrated. These diagnostic fragments of both red and black spruce were present at a frequency of over 0.95 in allopatric provenances, but at a lower frequency in some sympatric provenances (0.43–1.00). Natural populations of red spruce exhibiting typical red spruce phenotype contained black spruce diagnostic RAPD fragments and black spruces growing in bogs with typical bog black spruce morphology, contained red spruce-specific RAPD markers. Some major RAPD markers were cloned and sequenced. The results reveal an extremely high degree of identity between the random primer and the primer binding sites on the genome. Amplification of black and red spruce genomic DNA with designed primers flanking the species-diagnostic RAPD markers indicates that most of RAPD markers used to differentiate black spruce from red spruce are not species specific since these sequences were detected in several spruce species using a more sensitive detection method. Received June 17, 2002; accepted August 5, 2002 Published online: February 4, 2003     

凉水国家自然保护区天然红松种群遗传多样性在时间尺度上变化的cpSSR分析

应用叶绿体微卫星（cpSSR）技术对凉水国家自然保护区天然红松种群的遗传多样性在时间尺度上的变化进行了遗传分析。共选择121个树龄在1 581~1 900年间的红松个体,连续分为6个龄级。从6对叶绿体SSR引物筛选出两对引物,共检测到7个位点,其中多态位点5个,多态位点比率为71.43%。利用POPGENE分析软件计算了红松各龄级内的遗传多样性指数,分析了遗传多样性在红松各龄级间的变化。研究结果表明,凉水保护区内的红松遗传多样性在这320年间没有发生太大的波动,龄级间遗传多样性分化不明显,红松遗传多样性主要存在于龄级内。     

Application of ISSR, RAPD, and cytological markers to the certification of Picea mariana, P. glauca, and P. engelmannii trees, and their putative hybrids.

Picea glauca (white spruce) and P. engelmannii (Engelmann spruce) are so similar and integrated that it is impossible to distinguish between them and their hybrids using morphological characteristics. Although natural hybrids between P. glauca and P. mariana (black spruce) do not generally occur, even though the 2 species are sympatric in North America, a first-generation hybrid, called the Rosendahl spruce, has been reported in the literature. In this study, several inter-simple sequence repeat (ISSR) markers were developed, as were randomly amplified polymorphic DNA (RAPD) markers, to certify spruce trees and their hybrids. ISSR fingerprinting was more efficient than RAPD assay; it detected 70% polymorphic DNA markers among the spruce species analyzed, whereas RAPD fingerprinting detected only 53%. Species-diagnostic ISSR and RAPD markers differentiating P. glauca from P. engelmannii and P. mariana were cloned and sequenced. Molecular certification of the spruce samples analyzed confirmed that all the seeds from interior spruce populations were true hybrids of P. glauca and P. engelmannii. But the analysis of seeds derived from the putative Rosendahl spruce indicated that this tree is likely a pure P. glauca genotype, rather than a hybrid of P. glauca and P. mariana. These data were confirmed by cytological analyses. Further analysis, using a more sensitive DNA amplification method with designed primers flanking the species-diagnostic ISSR and RAPD markers, revealed that such sequences are not generally species-specific because they are present in other spruce species.     

Construction of a framework map for Pinus koraiensis Sieb. et Zucc. using SRAP, SSR and ISSR markers

Genetic linkage maps are essential for molecular breeding program. The first genetic linkage map of Pinus koraiensis, using an F1 progeny of 94 individuals, was constructed in the present paper. One hundred and twenty-two molecular markers were mapped onto 11 linkage groups, 1 triple and 8 pairs at the linkage criteria LOD 4.0. Among these markers, there were 96 sequence-related amplified polymorphism (SRAP) markers, 25 simple sequence repeat (SSR) markers, and 1 inter-simple sequence repeat (ISSR) marker. The consensus map gained covers 857.464 cM Kosambi (K) with an average marker spacing of 7.03 cM K. The presented map provides a basis and crucial information for future genomic studies of P. koraiensis, in particular for quantitative trait loci (QTL) mapping of economically important breeding target traits.     

Rapid identification of white-Engelmann spruce species by RAPD markers

Fragments of random amplified polymorphic DNA (RAPDs) were used as markers to distinguish Picea glauca (Moench) Voss (white spruce) and Picea engelmannii Parry (Engelmann spruce). These species and their putative hybrids are difficult to differentiate morphologically and are collectively known as interior spruce. Four oligodeoxynucleotide decamer primers showed species-specific amplification products between white spruce and Engelmann spruce. These fragments are highly conserved among seed lots and individual trees of each species from diverse geographic origins. The consistency and reproducibility of these species-specific amplification products were tested in more than two amplification reactions. Therefore, RAPD markers can provide genetic markers for easy and rapid identification of the specific genetic entry of these spruce species and their reported putative hybrids. According to the frequencies of the species-specific RAPD markers, it is possible to estimate the hybrid fraction, indicative of true introgression between the two species. These results are useful for quick identification of both species and their hybrid swarms at any stage in the sporophyte phase of the life cycle, for determining the occurrence and the magnitude of introgressive hybridization in an overlap zone between the two species, and for certification purposes in operational re-forestation and tree-improvement programs.