Establishment and Optimization of ISSR-PCR Reaction System for Picea crassifolia kom.

[Objective] The experiment aimed to determine the optimum ISSR-PCR reaction system of Picea crassifolia kom. [Method] Picea crassifolia kom. was used as material to select and optimize influencing factors of ISSR-PCR such as Mg2+, dNTPs, Taq DNA polymerase, template DNA, primers, annealing temperature. [Result] The optimum ISSR-PCR reaction system in 20 μl reaction system was consisted of 1 μl 10×buffer, 1.5 mmol/L Mg2+, 0.2 mmol/L dNTPs, 1.0 U Taq DNA polymerase, 40 ng template DNA, 0.6 μmol/L primers. According to gradient test of annealing temperature in optimum ISSR-PCR reaction system of Picea crassifolia kom, it was found that the optimum annealing temperature of UBC 818 was 54.2 ℃ and the annealing temperature was different for different primers.[Conclusion]The construction of ISSR-PCR reaction system provided technical basis for classification of germplasm resources, construction of genetic map, gene mapping of Picea crassifolia kom. through using ISSR technology.

Divergent responses of Picea crassifolia Kom.in different forest patches to climate change in the northeastern Tibetan Plateau

Global climate changes have significantly affected tree growth and forest structures and functions in some arid and semi-arid regions,which are becoming warmer and wetter.Due to natural factors such as climate and terrain,some tree species may form different forest patches at the edges of their distribution areas.However,how forest patches of various sizes respond to climate change is unclear.In this study,we collected 203 tree cores from six different sizes of forest patches at the edge of the distribution area of Picea crassifolia Kom.in the northeast Tibetan Plateau.And we used the dendrochronology method to study the response of tree growth and resilience in different forest patches to climate change from 1961 to 2020.We simultaneously measured the contents of nonstructural carbohydrates(NSC),total nitrogen and total phosphorus of tree needles.Our results showed that the growth of trees in small-and medium-size forest patches(0.8–18.6 ha)has increased significantly.The early growing season(May–July)minimum temperature was the most important climate factor driving the growth of small-and medium-sized patch trees.The early growing season maximum temperature was the most important climate factor that inhibited the growth of trees in the largest patches(362.8 ha).The growth of individual trees in medium forest patches was better and the correlation with annual minimum temperature,maximum temperature,precipitation,actual evapotranspiration,and palmer drought severity index was stronger.The higher NSC content,stronger photosynthesis,and higher nitrogen utilization efficiency in leaves might be one of the reasons for the better growth of trees in moderate forest patches.In extreme drought years,as the forest patch area increased,the overall trend of tree growth resistance showed a unimodal pattern,with the highest at a forest patch area of 7.1 ha,while the overall trend of tree growth recovery was opposite.Therefore,we should strengthen the management of trees in large forest patches to cope with climate change.

Differential response of radial growth and δ^(13)C in Qinghai spruce(Picea crassifolia) to climate change on the southern and northern slopes of the Qilian Mountains in Northwest China

Tree radial growth can have significantly differ-ent responses to climate change depending on the environ-ment.To elucidate the effects of climate on radial growth and stable carbon isotope(δ^(13)C)fractionation of Qing-hai spruce(Picea crassifolia),a widely distributed native conifer in northwestern China in different environments,we developed chronologies for tree-ring widths and δ^(13)C in trees on the southern and northern slopes of the Qilian Mountains,and analysed the relationship between these tree-ring variables and major climatic factors.Tree-ring widths were strongly influenced by climatic factors early in the growing season,and the radial growth in trees on the northern slopes was more sensitive to climate than in trees on the southern.Tree-ring δ^(13)C was more sensitive to climate than radial growth.δ^(13)C fractionation was mainly influenced by summer temperature and precipitation early in the growing season.Stomatal conductance more strongly limited stable carbon isotope fractionation in tree rings than photosynthetic rate did.The response between tree rings and climate in mountains gradually weakened as climate warmed.Changes in radial growth and stable carbon isotope fractionation of P.crassifolia in response to climate in the Qilian Mountains may be further complicated by continued climate change.

The type, position and age effect on the cutting reproduction of Picea crassifolia and its rooting mechanism in the Qilian Mountains

Picea crassifolia Kom, a perennial arbor spe- cies is recognized as one of the most adaptable plants found to date in Qilian Mountains. To explore the cutting reproduction technology of P. crassifolia and reveal its rooting mechanism, cuttings of P. crassifolia with different cutting types （softwood, hardwood and root）, positions （top, upper, middle and bottom） and ages （7, 10, 15, 20, 25 year-old） were cultivated in a field experiment. One- year old softwood and hardwood cuttings were collected from 7-, 10-, 15-, 20-, and 25 year-old healthy ortets to analyze the changes from endogenous hormones and organic nutrients. Results indicate that the softwood cut- tings （0.5-1.0 cm in diameter） from upper branches of 15 year-old ortets shows better growth performance by improving rooting indexes, including a significant increasein rooting rate and a decrease in basal rot rate. Concomi- tantly, increasing rooting quantity and root length also increased. It is noteworthy that the high rooting rate of P. crassifolia cuttings due to its ability to accumulate high concentrations of indole-3-acetic acid （IAA） and total carbon （TC） rather than abscisic acid （ABA） and total nitrogen （TN）. The rooting rate was mainly regulated by the IAA/ABA and TC/TN ratio. In summary, our results suggest that the softwood cuttings （0.5-1.0 cm in diameter） from upper branches of 15 year-old P. crassifolia can be considered as an effective strategy to improve cutting rooting rate, and the IAA/ABA and TC/TN ratio was one of the main factors limiting the cutting rooting rate of P. crassifolia.

干旱条件下苔藓层对祁连山青海云杉林土壤水分变化的缓冲作用

【目的】探讨干旱条件下苔藓层对青海云杉林土壤水分变化的影响,为理解气候变化背景下旱区土壤水分的变化规律及森林可持续管理提供依据。【方法】通过对干旱年和常水年有、无苔藓样地土壤水分的连续观测,分析不同干旱条件下苔藓层对土壤水分的影响。【结果】年际尺度上,常水年有、无苔藓样地平均土壤含水量分别为(29.5±6.1)%、(29.6±7.1)%,干旱年有、无苔藓样地平均土壤含水量分别为(22.3±4.7)%、(27.7±4.2)%;常水年苔藓层对青海云杉林土壤水分无显著影响(P>0.05),干旱年苔藓层会显著增加土壤含水量(P<0.05)。年内尺度上,不同月份苔藓层对土壤水分的作用存在差异,表现为生长季早期苔藓层对青海云杉林土壤水分的作用不显著(P>0.05),生长季中后期作用显著(P<0.05)。不同降雨条件下苔藓层对土壤水分的影响显著。晴天或小雨条件下,苔藓层对土壤水分作用不明显(P>0.05),中大雨条件下,苔藓层会显著增加土壤含水量(P<0.05)。【结论】苔藓通过增加入渗和抑制土壤蒸发,从而在干旱条件下对土壤水分变化起到缓冲作用。

祁连山东端林区青海云杉黄卷蛾发生规律调查及绿色防治药剂筛选

为了保护祁连山生态环境和实现青海云杉黄卷蛾绿色防治,观察了其生活习性,调查了发生地环境因子,并开展了不同药剂防治筛选试验。结果显示:在祁连山林区青海云杉黄卷蛾1年发生1代,发生受青海云杉分布地海拔和郁闭度影响,海拔2500~2800 m和郁闭度0.6以下林区发生较严重。使用苦参碱、印楝素、苏云金杆菌和多杀霉素防治后,均显著降低了虫口密度、林木受害率和死亡率,不同药剂中0.3%印楝素乳油的虫口密度、林木受害率和死亡率最低,综合防治效果最佳,2.5%苦参碱次之。

青海云杉硬枝扦插的激素、年龄和位置效应研究

为了解青海云杉插穗生根的外在因素和内在生理生化基础,采用网袋容器和自动间歇喷雾扦插繁殖技术,系统研究了青海云杉插穗母株年龄、部位、类型及激素处理等对插穗生根的影响,并采用高效液相色谱法测定了不同插穗类型内源激素吲哚乙酸(IAA)、脱落酸(ABA)、赤霉素(GA3)、玉米素(Z)、玉米素核苷(ZR)、异戊烯基腺嘌呤(IPA)、酚类化合物和糖类的含量。结果表明,以7年生母株枝条作穗条扦插繁殖效果最佳;一级侧枝的生根情况优于二级侧枝和顶枝穗条,树冠下部穗条生根优于上部枝条;激素处理能显著提高硬枝插穗的生根率,其中以IBA 200 m g/kg处理3 h的生根效果最好;邻苯二酸和儿茶酸2种酚类化合物可能对生根有抑制作用;枝条和针叶中三糖含量随母株年龄增加而增大,而葡萄糖含量却呈下降趋势;高含量的Z和ZR均有利于插条生根,IAA对生根无促进作用,IPA对插条生根可能具有一定的抑制作用,单一的某种激素对青海云杉插穗生根无明显影响。

青海云杉ISSR-PCR反应体系的建立与优化

[目的]确定适应青海云杉ISSR-PCR反应的最佳反应体系。[方法]以青海云杉(Picea crassifoliakom.)为材料,对影响其ISSR-PCR扩增结果的各因素(Mg2+、dNTPs、TaqDNA聚合酶、模板DNA、引物、退火温度)进行筛选和优化。[结果]青海云杉ISSR-PCR最佳反应体系为在20μl反应体系中包含1μl10×buffer,1.5 mmol/L的Mg2+,0.2 mmol/L的dNTPs,TaqDNA聚合酶1.0 U,模板DNA40 ng,引物0.6μmol/L。对青海云杉ISSR-PCR最佳反应体系的退火温度进行梯度试验发现,引物UBC818的最适退火温度为54.2℃,且最适退火温度因引物而异。[结论]该优化ISSR-PCR反应体系的建立,为今后利用ISSR技术进行青海云杉种质资源分类、遗传图谱构建和基因定位奠定了技术基础。

Pilodyn在青海云杉活立木基本密度预测中的应用

利用P ilodyn对青海云杉活立木南北方向进行探测,室内测定青海云杉木材各基本密度和早晚材管胞长,对青海云杉南北向P ilodyn值与木材各基本密度、早晚材管胞长、胸径间相关关系进行统计分析.结果表明,青海云杉P ilodyn值南向平均值为26.3 mm,北向平均值为27.3 mm,南北向P ilodyn平均值差异不显著,北向P ilodyn值与胸径间相关关系显著,青海云杉南北向P ilodyn值与整株木材基本密度均呈极显著的负相关,而与早晚材管胞长间都没有显著的相关关系.整株木材基本密度对青海云杉南向P ilodyn值的贡献率最大为60%,整株木材基本密度对北向P ilodyn值的贡献率最大为46%.

西宁周边山地青海云杉人工林地位级划分

为了给西宁周边山地及相似生态区提供青海云杉人工林立地质量评价标准和营造林技术指导,在西宁周边山地选择有代表性的27块样地,进行系统的优势木生长状况和立地因子调查。通过调查,分析主要立地因子对立地质量的影响,编制优势木高生长动态导向拟合方程。结果表明:指数回归方程y=-0.022x^3+1.410 7x^2-7.870 5x-64.97是树高地位级最适合的导向拟合曲线。以20 a为青海云杉基准林龄,确定按4个地位级评价立地质量。立地因子对立地质量的具体影响为低坡优于高坡,缓坡优于陡坡;阴坡优于半阴坡,半阴坡优于半阳坡,半阳坡优于阳坡,半阳坡和阳坡为贫瘠立地。

青海云杉嫁接技术的研究

该文以 1 986— 1 995年龙渠青海云杉无性系种子园嫁接生产实践为基础 ,对影响青海云杉嫁接效果的因素 ,如嫁接人员操作的熟练程度 ,嫁接时间 ,嫁接方法 ,嫁接时的天气状况 ,接穗的质量、年龄、来源、存储方法 ,砧木的年龄、所处环境等进行了综合分析 .结果表明 :通过严格培训嫁接人员 ,在川灌区选择 4月下旬、林区选择 5月中旬 ,无风、无雨的天气 ,用髓心形成层贴接法 ,将长 8～ 1 2cm、粗 5～ 1 0mm的 1年生接穗嫁接在 8～ 1 2年生的砧木上 ,可大大提高嫁接成活率、萌芽率 。

青海云杉种子园无性系结实性状遗传变异研究

[目的]研究无性系结实性状的遗传变异规律.[方法]利用青海云杉一代无性系种子园中筛选出的18个优良无性系90个分株材料,研究青海云杉无性系结实性状的遗传变异和不同树冠层的结实量.[结果]青海云杉无性系间的结实能力差异明显,其无性系重复力较高,青海云杉无性系结实母树球果主要集中在树冠中上部的冠层分枝上,青海云杉无性系球果主要分布在树冠上部,各部位的球果量从大到小依次为上部、中部、下部.球果量在树冠中部、上部、下部分别占单株球果总量的86.43%、10.37%和3.20%.通过性状相关分析,青海云杉单株结实量与树冠高呈极显著正相关,下部结实量与冠幅呈极显著负相关,中部结实量、单株球果量和单果鲜重3个结实性状与无性系生长性状间均无显著的相关关系.[结论]该研究可为青海云杉无性系种子园的营建及经营管理提供科学依据.

青海云杉半同胞家系遗传变异及二代优树选择

对甘肃省小陇山沙坝试验基地9年生40个青海云杉家系子代测定林树高、新梢、冠幅进行了测定和分析。结果表明,青海云杉家系间各性状差异均达显著水平。青海云杉各性状中家系遗传力均大于单株遗传力,各家系间树高、新梢、冠幅三者之间表型相关均为极显著相关。以家系树高育种值为选择指标,共选出8个优良家系。采用配合选择最佳线性预测(BLUP)估算单株育种值,选出来自21个家系的102个优良单株,其树高比群体均值高出36.25%,期望遗传增益是27.97%。说明配合选择具有兼顾高遗传增益和广遗传基础的优点。这些优良家系和单株可作为营建青海云杉二代种子园试验林的材料。

青海云杉种子园树木生长相关性分析

青海云杉种子园是提供具有优良遗传品质的更新造林用种的主要基地。青海林木良种育种工作比较薄弱,我们应用生长相关性分析理论,开展青海云杉种子园树木高生长与胸径以及树冠之间的相关性分析来拟定青海云杉生长过程,并根据生长变量自身的数据序列,建立起青海云杉树高生长与胸径生长、胸径生长与树冠的动态预测模型。结果表明:应用线型分析理论建立的树高生长量模型和胸径总生长量模型以及树冠生长量模型,精度相当高,能够准确地预测青海云杉树高生长的未来趋势。

甘肃祁连山青海云杉种群数量动态的初步研究

用生存分析和静态生命表研究方法,在祁连山国家级自然保护区调查了16个(20 m×20 m)样地,分析青海云杉(Picea crassifolia Kom.)种群数量动态和年龄结构.结果表明,不同种群年龄结构不同,属顶级群落的苔藓-青海云杉林、草类-青海云杉林表现为衰退型种群;灌木-青海云杉林、马先蒿-青海云杉林为增长型种群.1～5龄级存活曲线较陡,种群死亡率较高;5龄级以后存活曲线相对平缓,死亡率较低.采用指数方程和幂函数对生存曲线的类型进行检验,苔藓-青海云杉林、草类-青海云杉林和标准化青海云杉林的生存曲线符合DeeveyⅢ型曲线;灌木-青海云杉林、马先蒿-青海云杉林的生存曲线符合DeeveyⅡ型曲线.种群数量变化的回归分析和青海云杉生态寿命与环境因子相关性分析表明,青海云杉种群存活数变化和生态寿命与种群密度有关.

粗枝云杉、青海云杉叶锈病病原及转主寄主研究

对白龙江林区云杉、青海云杉叶锈病的病原形态、转主寄主与寄主病原交互隔离接种后认为 ,2种云杉叶锈病病原均为祁连金锈菌 Chrysomyxa qilanensis Wang.Wu et L .粗枝云杉叶锈病病原转主寄主为山光杜鹃 Rhododendron oreodoxa Franch,青海云杉叶锈病病原转主寄主为青海杜鹃 Rh.przeowalskii

青海云杉种实害虫化学防治技术

青海云杉(Picea crassofpla kom.)种实害虫的优势种是云杉球果小卷蛾(Pseudotomoides strobilellus),严重为害区平均虫果率达81.6%,是化学防治的主要对象.试验分别对面积较大、郁闭度高、树体高大的林分,面积小、郁闭度低,树体较矮的母树林或种子园及虫害程度中等或轻微的稀疏、低矮林分进行了防治试验,取得了较好的效果.

青海云杉半同胞子代苗期生长性状遗传分析

以青海云杉无性系种子园内30个青海云杉半同胞家系为研究对象,在张掖市肃南县塔尔沟苗圃开展苗期试验,测定5a生苗期苗高、地径、新梢长、侧枝数等生长性状。分析结果表明:(1)青海云杉半同胞子代苗家系间苗高、地径、新梢长存在极显著差异;遗传变异系数较高,均在19.76%以上,苗高、地径、新梢长、侧枝数指标的家系遗传力均较大,均在0.830以上;苗高与地径、新梢长间存在极显著正相关,地径与新梢长、侧枝数存在极显著正相关;(2)以苗高为首要选择指标,按照1倍标准差选择出158#、106#、160#、154#、109#、119#6个优良家系,各性状值高于30个家系的平均值。

青海云杉育种资源遗传评价

【目的】了解青海云杉无性系种子园的遗传多样性及无性系之间的亲缘关系,为青海云杉遗传改良策略的制定及高世代种子园的建立提供理论基础.【方法】利用SSR分子标记对甘肃省张掖龙渠青海云杉无性系种子园来自9个种源地共108个无性系进行遗传多样性及种源间、无性系间的亲缘关系分析.【结果】10对SSR引物共扩增出50个多态位点,平均有效等位基因数为2.4842,平均期望杂合度为0.5154,Nei多样性指数为0.5129,Shannon信息指数为1.0065.种源间的遗传分化系数为0.2816,基因流为0.6378,表明种源间存在较高程度的遗传分化,分子方差分析(AMOVA)结果表明遗传变异主要存在于个体间(84%).基于Nei’1983遗传距离进行UPGMA聚类,9个种源分为2大类群,108个无性系基本按照种源聚类.【结论】张掖龙渠青海云杉无性系种子园具有较高的遗传多样性水平,可为高世代种子园的建设提供丰富的原材料.

西宁市青海云杉人工林健康评价研究

本文以西宁市南北山20-30a生青海云杉人工林为研究对象,选定树高年生长量、胸径年生长量等10个评价指标进行赋值和无量纲化处理,并用变异系数法确定其权重,通过参考健康度指标(HI)构建评价模型和正态等距法确定健康等级标准,最终分析并得出西宁市南北山青海云杉人工林健康水平。研究结果表明,各健康等级的取值范围为:不健康[0.22,0.33]、中健康[0.34,0.45]、亚健康[0.46,0.57]、健康[0.58,0.69];人工林中达到健康水平的有17%、亚健康的占42%、中健康的占28%、不健康的占13%,从隶属度最大的原则上分析,西宁市南北山青海云杉人工林以亚健康状态为主。