利用ISSR标记对12种五针松亲缘关系的研究

采用ISSR 标记技术,对12 种五针松的亲缘关系进行分析。利用筛选出的12 个ISSR 引物共检测到117 个位点,多态条带比率(PPB)在9.40%～33.33 %之间,遗传分化最高的是偃松,最低的是柔枝松。12 种五针松的基因多样性(Ht)为26.21%,其中种内基因多样性(Hs)为7.66%,种间基因多样性(Dst)为18.55%,五针松种间变异占总变异的70.78%。遗传距离聚类,将12 种五针松分为2 个类群,第一类群包括乔松、华山松、海南五针松、华南五针松、北美乔松、山白松和云南五针松;第二类群包括美国白皮松、偃松、、柔枝松、西伯利亚红松和红松。图1 表3 参6。     

无患子天然居群遗传多样性研究

[目的]通过我国无患子主要分布区的居群样本,研究无患子天然居群的遗传多样性和遗传结构.[方法]采用ISSR分子标记技术,利用12条ISSR引物分析18个天然居群的265株个体样本.[结果]表明无患子遗传多样性水平较高,物种和居群水平上的多态位点百分率 (PPB)分别为95.37%和57.82%,Shannon's信息指数(I)分别为0.256 9和0.199 8,Nei's遗传多样性指数(H)分别为0.390 9和0.298 0.AMOVA分析表明,18个居群间出现一定程度的遗传分化,且遗传变异主要发生在居群内.UPGMA聚类和Mantel检验结果表明,18个天然居群可分为2大组群,且居群间的地理距离与遗传距离之间不存在显著相关性(r=0.066 7,P=0.541 7>0.05).[结论]无患子以自交为主,其天然居群遗传多样性丰富,居群内的遗传多样性高于居群间.研究结果可为无患子育种策略的科学制定和种质资源的有效保护及利用提供理论依据.     

Genetic variation and division of Pinus sylvestris provenances by ISSR markers

Inter Simple Sequence Repeat (ISSR) was used to detect genetic variation among nine provenances, including 135 individuals. A total of 108 loci were amplified using 10 random primers. The differentiation of the percentage of polymorphic bands (PPB) among different provenances was evident, ranging from 27% to 54%, of which Honghuaerji provenance had the highest PPB and Kalunshan provenance had the lowest PPB. Shannon's Information index (I) at species level was 0.1581 and Nei's gene diversity (h) was 0.2393. Coefficient of gene differentiation (G_ST) calculated by Popgene was 0.3965, these results indicated that majority of genetic variation (60.35%) was found within provenances. According to dendrogram amongpinus sylvestriv provenances, nine provenances were divided into two provenance areas, namely Daxing's and Xiaoxing'an Mountains provenance area and Hulunbeier provenance area. Foundation item: This study was supported by National Scientific and Technological Brainstorm Project. Biography: LI Hui-yu (1978-), female, Ph.D. candidate, Northeast Forestry University, Harbin 150040, P. R. China. Responsible editor: Song Funan     

花吊丝竹居群遗传多样性的ISSR分析

[目的]研究花吊丝竹居群遗传多样性和遗传结构,为种质资源有效利用和良种选育提供理论指导。[方法]利用12条ISSR引物对48份种质(共3个居群)花吊丝竹居群进行遗传多样性和遗传距离分析。[结果]共检测到124个位点,其中,多态性位点为102个,种质和居群水平上的多态位点百分比(PPB)分别为82. 26%和50. 27%,Ne’基因多样性指数(He)分别为0. 220 4和0. 206 6,Shannon’s信息指数(I)分别为0. 349 4和0. 300 5,表明花吊丝竹居群间存在中等水平的遗传变异。根据Nei’s遗传多样性计算出不同居群间分化水平(Gst)=0. 163 3,表明16. 33%的遗传变异存在于居群间,居群内的遗传变异为83. 67%。居群间的基因流Nm为2. 562 1,表明花吊丝竹居群间存在较大基因流,很大程度减少居群间遗传差异。基于遗传距离的UPGMA聚类结果表明,48份种质可分为3组,3个居群可分为2组,居群间地理距离与亲缘关系无显著相关性。[结论]虽然花吊丝竹主要靠营养生殖来繁衍后代,其居群遗传多样性较丰富,且居群内遗传多样性大于居群间。此外,福建居群遗传多样性明显高于广西和广东地区居群。     

Comparison of species diversity betweenLarix gmelini pure forest andLarix gmelini-betula platyphylla mixed forest in Daxing'an Mountains

Based on the investigation data about the species diversity in Daxing'an Mountains, the difference of species diversity indexes betweenLarix gmelini forest andLarix gmelini-Betula platyphylla forest was analyzed by the Simpson index (D), Shannon-Wiener index (H), Pielou index (J _sw &J _si), Alatalo index (E _a) and Species richness index (S). The result showed that the species diversity index ofLarix gmelini-Betula platyphylla forest was much higher than that ofLarix gmelini forest in this region. AndLarix gmelini-Betula platyphylla forest was beneficial to maintain the species diversity in Daxing'an Mountains region. Biography: SONG Guan-ling (1969-), female, Master, assistant professor in Zibo College, Zibo 255000, P.R. China. Responsible editor: Zhu Hong     

长白赤松和红松土壤微生物活性对高浓度CO2的响应

Responses of soil microbial activities to elevated CO₂ in experiment sites ofPinus sylvestriformis andPinus koraiensis seedlings were studied in summer in 2003. The results indicated the number of bacteria decreased significantly (p<0.05) under elevated CO₂ forPinus sylvestriformis andPinus koraiensis. Amylase and invertase activities in soil increased forPinus sylvestriformis and decreased forPinus koraiensis with CO₂ enrichment compared with those at ambient (350 μmol·mol⁻¹). The size of microbial biomass C also decreased significantly at 700 μmol·mol⁻¹ CO₂. Bacterial community structure had some evident changes under elevated CO₂ by DGGE (Denaturing Gradient Gel Electrophoresis) analysis of bacterial 16S rDNA gene fragments amplified by PCR from DNA extracted directly from soil. The results suggested that responses of soil microorganisms to elevated CO₂ would be related to plant species exposed to elevated CO₂. Foundation item: The study was supported by Major State Basic Research Development Program of China (2002CB412502) and the Knowledge Innovation Project from Chinese Academy of Sciences (KZCX1-SW-01-03). Biography: JIA Xia (1975), female, Ph. D. candidate of Institute of Applied Ecology, Chinese Academy of Sciences, Shenyang 110016, P. R. China. Responsible editor: Song Funan     

Genetic diversity analysis in a seed orchard of <Emphasis Type="Italic">Eucalyptus tereticornis</Emphasis>

India is the largest grower of Eucalyptus with an area of 3.943 million hectares under plantations and E. tereticornis is the predominant species in the plains of southern India with an average productivity of 12–25 m³ ha⁻¹ year⁻¹. With the aim to establish seedling seed orchards of the species, seed lots of fifteen provenances were imported from Australia and a trial was laid. In the present study the genetic diversity existing in the seed orchard was estimated using ISSR–PCR. Seven ISSR primers amplified 663 amplicons in the size ranging from 255 to 2,711 bp. The total number of polymorphic bands varied from 59 to 123 with 100% polymorphic banding profiles. The average gene diversity (Hj) of all the provenances ranged from 0.0589 to 0.1109 and the total gene diversity estimated was low (H _T = 0.130) when compared to the earlier reports from other eucalypts species. Analysis of Molecular Variance partitioned the ISSR variation into inter and intra population components. The inter population component accounted for 55.2% of the variation and the intra population component accounted for 46.3% (P < 0.001). A neighbour-joining analysis was done using the dissimilarity matrix to determine the aggregation of the individuals into clusters. Existence of population structure among the populations was revealed in STRUCTURE analysis but geographical region based clustering was not observed. The assessment of intra and inter genetic variation documented in the present study suggests that, along with the phenotypic traits, knowledge about genetic diversity measured at the DNA level in individuals of seed orchards provide an objective guide for selective culling of trees for maintaining optimal diversity for enhanced genetic gains.     

Vitality and storage condition of Syringa pollen

The fresh pollen vitality,the effect of different storage conditions on the pollen vitality,and the difference of vitality among the species of seven species of Syringa were determined in Shenyang,China.The results indicated that the pollen vitality (81.5%) of Syringa villosa was the highest among the seven tested species,followed by S.microphylla and S.meyeri,and that of S.oblata var.affinis was the lowest.The low temperature was the best condition for storage of pollen of Syringa,and the most proper temperature for the storage was 0-2℃.The storability of S.microphylla was the best of all,and it could be stored over 60 days at the temperature of 0-2℃,next was S.villosa and S.meyeri.     

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.     

Effect of water stress on N2O emission rate of 5 tree species

The N₂O emission rates, photosynthesis, respiration and stomatal conductance of the dominant tree species from broadleaf/Korean pine forest in Changbai Mountain were measured by simulated water stress with the closed bag-gas chromatography. A total of five species seedlings were involved in this study, i.e.,Pinus koraiensis Sieb. et Zucc,Fraxinus mandshurica Rupr,Juglans mandshurica Maxim,Tilia amurensis Rupr, andQuercus mongolica Fisch. ex Turcz.. The results showed that the stomatal conductance, net photosynthetic rate and N₂O emission of leaves were significantly reduced under the water stress. The stoma in the leaves of trees is the main pathway of N₂O emission. N₂O emission in the trees mainly occurred during daytime. N₂O emission rates were different in various tree specie seedlings at the same water status. In the same tree species, N₂O emission rates decreased as the reduction of soil water contents. At different soil water contents (MW, LW) the N₂O emission rates ofPinus koraiensis decreased by 34.43% and 100.6% of those in normal water condition, respectively. In broadleaf arbor decreased by 31.93% and 86.35%, respectively. Under different water stresses N₂O emission rates in five tree species such asPinus koraiensis, Fraxinus mandshurica, Juglans mandshurica, Tilia amurensis, andQuercus mongolica were 38.22, 14.44, 33.02, 16.48 and 32.33 ngN₂O·g⁻¹DW·h⁻¹, respectively. Foundation item: This project was supported by the National Natural Science Foundation of China (No. 30271068), the grant of the Knowledge Innovation Program of Chinese Academy of Sciences (KZ-CX-SW-01-01B-10), and the Special Funds for Major State Basic Research Program of China (No. G1999043407) Biography: Wang Miao (1964-), male, associate professor in Institute of Applied Ecology, Chinese Academy of Science, Shenyang 110016, P. R. China. Responsible editor: Song Funan     

Genetic diversity and genetic differentiation of natural Pinus koraiensis population

Genetic diversities and genetic differentiations of the four Pinus koraiensis populations （Gaofeng Forestry Farm at Tangwanghe in Yichun City, Erdaobaihe Town in the Changbai Mountains, Shcngshan Forestry Farm in Heihe City, in China, and the suburb of Vladivostok City in Russia） were analyzed by using ISSR PCR technique. The results of 15 primers amplification showed that the ratio of polymorphic site of P koralensis population was 60.7%. Euch primer had 3.6 polymorphic sites, The diversity levels of the four P. koraiensis populations were rather higher compared with those of other Pinaceae species. The genetic diversity levels of t： koraiensis populations grown in the center region were higher than those grown in the edge zone. The genetic diversity of r3 koraiensis mainly came from the interiot of the population, accounting for 73% of total genetic diversity. There were no positive correlation between genetic distances and geographical distances for the four P koraiensis populations. The gradual decrease of natural distribution region of P. koraiensis was due to anthropic destroy and environmental factors （i.e. fire and wind throw）, rather than the lower genetic diversity.     

毛红椿天然群体遗传多样性及取样策略探讨

[目的]研究毛红椿天然群体遗传多样性取样策略,为其种质资源收集、保存和遗传多样性保护等提供参考依据。[方法]利用8对微卫星分子标记进行毛红椿天然群体遗传多样性和空间自相关分析,综合制定其天然群体合理取样策略。[结果]毛红椿天然群体等位基因数平均为7.5个,期望杂合度(H_e)和多态性信息指数(PIC)均值分别为0.643 7和0.636 0,基因分化系数(G_(ST))均值为0.290 7。在遗传多样性取样策略方面,提出了根据毛红椿群体基因分化系数来确定取样群体遗传变异所占总变异比例的运算公式为1-(G_(ST))~(n-1),其中,n为取样群体的数量。当取样群体达到4个时,基本上能包括该树种97.5%的遗传变异;同时确定了目标群体的选择方法,应选择与其它群体间基因分化系数均值较大的4个群体,即贵州册亨(CH)、浙江遂昌(SC)、浙江仙居(XJ)和云南师宗(SZ)。通过构建云南宾川(BC)、云南师宗(SZ)和江西宜丰(YF)群体内取样单株数量与基因多样性和等位基因之间的捕获曲线,确定了群体内取样单株数量应达到15个以上;毛红椿天然群体内300～520 m范围内的单株间存在相似关系,超出此范围个体间差别较大,说明在进行群体内单株取样时,单株间距应大于520 m。[结论]取样群体数量、群体间遗传分化系数、群体内单株数量以及单株间距离等影响了毛红椿取样群体的遗传多样性。毛红椿天然群体遗传多样性取样策略为取样群体4个、每个群体最少取样15个单株,单株间距大于520 m。     

新疆额尔齐斯河流域杨属植物种间关系的SSR分析

[目的]为探讨额河杨和银灰杨天然杂种的起源机制,[方法]应用18对SSR标记,从分子水平上对新疆额尔齐斯河流域杨属植物的种间关系进行分析研究。[结果]表明:(1)SSR系统发育树将整个流域天然杨属植物分为两大类群,即黑杨派和青杨派为一类,白杨派为一类;(2)白杨派派内系统聚类图显示,银白杨、欧洲山杨、银灰杨三个树种均有较大的遗传分化,特别是杂种银灰杨似乎更大;(3)黑杨派和青杨派的UPGMA分类图显示,青杨派和黑杨派分属于2个分支,其中,青杨派内部分化相对简单,分为2支,均为典型的苦杨;黑杨派内部的分化较为复杂,可分为4类,包括典型的欧洲黑杨、额河杨和回交子代。[结论]杂种额河杨具有更多的欧洲黑杨的遗传成分,因此,将额河杨放到黑杨派是正确的。     

Evaluation Of Genetic Diversity Using Inter-Simple Sequence Repeat Markers And Effect On The Severity Of Leaf Blight Disease Of Teak (Tectona Grandis L.)

ABSTRACT

Leaf blight is an important disease of teak which reduces photosynthetic rate and ultimate timber yield in infected trees. Symptomatic leaf samples were randomly collected from three forest reserves, Onigambari, Olokemeji and Omo in Southwest Nigeria. Genomic DNA from the samples was extracted using Cetyl Trimethyl Ammonium Bromide (CTAB) method and amplified with Inter Simple-Sequence Repeat (ISSR) markers to evaluate genetic diversity. Dendrogram analysis using UPMA procedure separated the accessions into 12 major groups with bootstrap values ranging from 51 to 62. The constructed dendogram showed that accessions from Onigambari forest reserve were genetically similar to those from the two other reserves. Principal component analysis (PCA) showed that the first, second and third axes accounted for 79.19%, 85.41%, and 88.11%, respectively, of the total molecular variation. All the four ISSR markers showed 100% polymorphism which induced the amplification of 48 alleles across the 39 teak accessions evaluated. Polymorphic information content (PIC) obtained from ISSR markers ranged from 0.6038 to 0.8886, while genetic diversity varied from 0.6288 to 0.8947. The inter-population genetic diversity and allelic richness observed in the study could be employed in breeding for resistance against leaf blight disease of teak.     

10个南方皂荚群体遗传多样性的AFLP分析

[目的]研究南方皂荚的遗传多样性和遗传结构现状,为制定有效的保护策略和育种策略提供理论依据。[方法]采用扩增片段长度多态性技术对供试的215份皂荚的遗传多样性及遗传距离进行分析。[结果]表明:检测到1 782个位点,其中,有1 389个为多态性位点,多态位点百分率为77.94%;皂荚各群体Shannon’s信息指数平均为0.256;Nei’s多样性指数平均为0.168,表明皂荚群体的遗传多样性偏低,皂荚群体间存在着一定的遗传分化;根据遗传距离UPGMA聚类分析,将10个群体划分为4大类。由遗传分化系数和分子方差分析结果发现,群体内的变异是皂荚遗传变异的主体,74.79%的变异来自群体内。[结论]皂荚群体的遗传多样性与遗传结构的形成不仅与其分布广泛、种子特性及生活史有关,而且与人为的砍伐、引种、生境片段化等因素有重要关系。基于上述结果提出了皂荚的保护策略。     

Genetic diversity ofKandelia candel andBruguiera gymnorrhiza in the Southwest Islands, Japan

The genetic diversity of the mangrove tree speciesKandelia candel andBruguiera gymnorrhiza in the Southwest Islands of Japan was investigated. These islands are located at the periphery of the distribution area of the two species, and their populations are relatively small. Allozyme analyses of 17 loci inK. candel and 13 loci inB. gynmorrhiza revealed very little genetic variation in both species. At the species level, the proportion of polymorphic loci was 4.2%, the average effective number of alleles per locus was 1.02, and the average expected heterozygosity was 0.012 inK. candel. The corresponding figures forB. gymnorrhiza were 9.0%, 1.06 and 0.035, respectively. However, the coefficients of genetic differentiation among the populations were high (G _ST=0.165 forK. candel and 0.253 forB. gymnorrhiza). The genetic variation and habitat area ofK. candel on Amami Island is greater than on the islands Okinawa and Iriomote. The genetic variation and habitat area ofB. gymnorrhiza is greater on Iriomote Island than on the islands Okinawa and Amami. The level of genetic variation in both of the species might be related to their population sizes.     

红豆树优树子代遗传多样性及与生长相关性分析

[目的]研究红豆树优树自由授粉子代遗传多样性及其对生长的影响,比较天然居群子代和孤立木子代遗传多样性差异,揭示子代遗传多样性的变化规律及天然居群在子代遗传多样性维持中的作用,为红豆树遗传保育和优异种质挖掘提供科学依据。[方法]以来自浙、闽、赣、川等26个红豆树优树自由授粉家系为研究对象,利用11对SSR引物对765个子代群体进行遗传多样性评价,同时分析子代遗传多样性参数与种子、生长性状的相关性。[结果](1)红豆树优树子代群体具有较高的遗传多样性,有效等位基因数为7.766个,观测杂合度(H_O)和期望杂合度(H_E)分别为0.469和0.865。(2)除SSR8外,其余位点的观测杂合度均小于期望杂合度,表明子代群体绝大多数位点处于杂合子缺失状态。(3)红豆树不同家系的遗传多样性存在明显差异,12号家系的遗传多样性水平最高,8号家系则最低。(4)比较发现,天然居群子代的遗传多样性显著或极显著地高于孤立木子代。(5)F统计量和分子方差分析(AMOVA)均表明,红豆树优树子代群体的遗传变异主要存在于家系内,家系间的遗传分化相对较小。(6)相关性分析发现,子代遗传多样性参数与种子性状、子代年高生长量呈显著正相关(r=0.378~0.527)。[结论]较大的红豆树天然居群在维持其子代较高遗传多样性中发挥了重要作用,子代遗传多样性显著影响苗木生长,这为红豆树遗传保育和优良家系选择提供了理论依据。     

Assessment of Genetic Diversity in Feronia limonia (L.) Swingle Using Inter Simple Sequence Repeats

ABSTRACT

Genetic variation was studied in threatened natural populations of Feronia limonia (Rutaceae). Samples were collected from 3 populations located in the Aravalli mountain range of south-east Rajasthan, India. Using 25 Inter Simple Sequence Repeat (ISSR) primer combinations, significant diversity was characterized among the individuals. Analysis of Molecular Variance (AMOVA) revealed that diversity is partitioned mainly within population components (80%). F statistics showed that low but significant amount of gene flow is taking place between populations (φ_ST = 0.21, P < .001). The study highlights the utility of ISSRs in assessing diversity of populations that are fragmented or limited in size.     

Assessment of morphological and genetic diversity in <Emphasis Type="Italic">Gmelina arborea</Emphasis> Roxb.

Gmelina arborea is an important timber-yielding tree that grows naturally in the tropical and subtropical regions of Southeast Asia and has also been introduced as a plantation species outside these regions. Genetic diversity in this tree species was observed in stone/seed-related traits and in vitro responses of cultured nodal segments from plants of eight different populations representing natural forests, fragmented forests and plantations. Variance analysis showed significant differences between populations for these traits. However, it was not possible to separate the different populations using these traits by multivariate analysis, even after environmental variation was reduced over six subcultures. Genetic diversity was therefore analysed using molecular markers. Inter-simple sequence repeat (ISSR) primers yielded 95% polymorphic loci among the eight populations and UPGMA analysis enabled separation of these populations on the basis of their genetic distances. Diversity was also analyzed using population genetics parameters like Nei’s genetic diversity and gene differentiation. Nei’s genetic diversity was 0.29 between populations and 0.11 within populations. AMOVA analysis indicated 41 and 59% within- and between-population genetic diversity, respectively. Mantel test suggested that genetic differentiation between six Indian populations was positively correlated to geographic distance (r = 0.626, P = 0.029). Assessment of the genetic variation in G. arborea populations is an important step in selection of conservation strategies for this species since diversity forms the basis for species adaptation.     

Response of seedlings of different tree species to elevated C02 in Changbai Mountain

Eco-physiological responses of seedlings of eight species,Pinus koraiensis, Picea koraiensis, Larix olgensis, Populus ussuriensis, Betula platyphylla, Tilia amurensis, Traxinus mandshurica andAcer mono from broadleaved/Korean pine forest, to elevated CO₂ were studied by using open-top chambers under natural sunlight in Changbai Mountain, China in two growing seasons (1998–1999). Two concentrations of CO₂ were designed: elevated CO₂ (700 μmol·mol⁻¹) and ambient CO₂ (400 μmol·mol⁻¹). The study results showed that the height growth of the tree seedlings grown at elevated CO₂ increased by about 10%–40% compared to those grown at ambient CO₂. And the water using efficiency of seedlings also followed the same tendency. However, the responses of seedlings in transpiration and chlorophyll content to elevated CO₂ varied with tree species. The broad-leaf tree species were more sensitive to the elevated CO₂ than conifer tree species. All seedlings showed a photosynthetic acclimation to long-term elevated CO₂. Foundation item: The project was supported by National Key Basic Development of China (G1999043400) and the grant KZCX 406-4, KZCX1 SW-01 of the Chinese Academy of Sciences Biography: WANG Miao (1964-), maie, associate professor in Institute of applied Ecology, Chinese Academy of Sciences, Shenyang 110016, P. R. China. Responsible editor: Song Funan