火炬松(Pinus taeda L.)合子胚愈伤组织的器官发生和植株再生

以火炬松(Pinus taeda L.)的成熟合子胚为外植体在附加NAA和BA的TE培养基上诱导产生了淡黄色、疏松、有光泽的颗粒状愈伤组织。经过愈伤组织的保持和增殖培养及不定芽原基的诱导培养后,进行了不同激素、低温处理和蔗糖浓度对不定芽分化的影响实验。结果表明,在附加0.5 mg·L^(-1)IBA和2 mg·L^(-1)BA的TE培养基上,愈伤组织上的不定芽分化频率最高达62.15%。不定芽分化的最佳低温处理时间是5—6周,最佳蔗糖浓度是25—30 g·L^(-1)。不定芽经伸长培养后取高于1cm的小苗用于生根。在附加IBA、BA和GA_3的TE培养基上不定芽的生很频率最高达46%。

Agrobacterium-mediated transformation and assessment of factors influ-encing transgene expression in loblolly pine (Pinus taeda L.)

This investigation reports a protocol for transfer and expression of foreign chimeric genes in loblolly pine (Pinus taeda L.). Transformation was achieved by co-cultivation of mature zygotic embryos with Agrobacterium tumefaciens strain LBA4404 which harbored a binary vector (pBI121) including genes for β-glucuronidase (GUS) and neomycin phosphotransferase (NPTII). Factors influencing transgene expression including seed sources of loblolly pine, concentration of bacteria, and the wounding procedures of target explants were investigated. The expression of foreign gene was confirmed by the ability of mature zygotic embryos to produce calli in the presence of kanamycin, by histochemical assays of GUS activity, by PCR analysis, and by Southern blot. The successful expression of the GUS gene in different families of loblolly pine suggests that this transformation system is probably useful for the production of the genetically modified conifers.

Genetic transformation of Pinus taeda by particle bombardment

A protocol is presented for genetically engineering loblolly pine (Pinus taeda L.) using particle bombardment. This protocol enabled the routine transformation of loblolly pine plants that were previously difficult to transform. Mature zygotic embryos were used to be bombarded and to generate organogenic callus and transgenic regenerated plants. Plasmid pB48.215 DNA contained a synthetic Bacillus thuringiensis (B.t.) cryIAc coding sequence flanked by the double cauliflower mosaic virus (CaMV) 35S promoter and nopaline synthase (Nos) terminator sequences, and the selectable marker gene, neomycin phosphotransferase II (nptII) controlled by the promoter of the nopaline synthase gene was introduced into loblolly pine tissues by particle bombardment. The transformed tissues were proliferated and selected by kanamycin resistance conferred by the introduced NPTII gene. Shoot regeneration was induced from the kanamycin-resistant callus, and transgenic plantlets were then produced. The presence of the introduced genes in the transgenic loblolly pine plants was confirmed by polymerase chain reactions (PCR) analysis, by Southern blot analysis, and insect feeding assays. The recovered transgenic plants were acclimatized and then established in soil.

Influences of antibiotics on plantlet regeneration via organogenesis in loblolly pine (Pinus taeda L.)

Three antibiotics ampicillin, carbenicillin, and cefotaxime were evaluated for their effects on induction, growth, and differentiation of organogenic calli, as well as rooting of regenerated shoots of three loblolly pine (Pinus taeda L.) genotypes. Of the antibiotics administered, cefotaxime maximally increased the frequency of callus formation and growth rate of organogenic calli, carbenicillin maximally increased the frequency of shoot regeneration and the average number of adventitious shoots per piece of organogenic callus, ampicillin maximally decreased the rooting frequency of regenerated shoots and mean number of roots per regenerated shoot, in comparison with antibiotic-free media. Compared with the control, ampicillin minimally increased the frequency of callus formation, cefotaxime minimally increased the frequency of shoot regeneration, and carbenicillin mini-mally decreased the rooting frequency of regenerated shoots in three loblolly pine genotypes tested. All three antibiotics in-creased the frequencies of callus formation and shoot regeneration, and reduced the rooting frequency of regenerated shoots suggested that the establishment of an efficient Agrobacterium tumefaciens-mediated transformation protocol for stable integra-tion of foreign genes into loblolly pine need to select a suitable antibiotic. This investigation could be useful for optimizing genetic transformation of conifers.

速生抗病火炬松Pinus taeda苗的磁化培育和早期筛选技术研究

研究表明,利用磁场处理火炬松种子能显著提高种子发芽率、苗木抗病性和促进苗高生长,并以浸种后的冷藏期处理效果最好;利用病原菌的毒素粗提液可进行抗病苗的早期筛选。

不同松树树种段木栽培茯苓产量对林分郁闭度的响应

【目的】探明林分郁闭度和不同松树树种段木组合栽培对茯苓产量的影响,为提高茯苓栽培产量提供科学依据。【方法】以茯苓为研究对象,选择低、中、高3种郁闭度林分,分别采用湿地松、马尾松和火炬松3种松树段木栽培茯苓,考察松树不同松种与林分郁闭度组合处理茯苓菌核个数、单个鲜重和总鲜重,通过方差分析和多重比较筛选树种与林分郁闭度的最佳搭配方案。【结果】湿地松段木栽培茯苓水溶性多糖含量为0.49%,马尾松和火炬松段木栽培均为0.26%。湿地松和马尾松段木栽培,中郁闭度林分茯苓单个鲜重和总鲜重显著高于低郁闭度;火炬松段木栽培,高郁闭度林分茯苓总鲜重低于中郁闭度且显著低于低郁闭度。【结论】湿地松、马尾松和火炬松3种松树段木栽培的茯苓综合品质良好,其中,以湿地松段木栽培茯苓的可溶性多糖含量最高。3种郁闭度林分环境下最佳段木栽培搭配方式:低郁闭度林分+火炬松段木,中/高郁闭度林分+湿地松或马尾松段木。

Studies on Parameters Affecting Embryogenesis of Protoplasts Isolated from Suspension of Embryogenic Cells in Loblolly Pine

Protoplasts of embryogenic suspension cells of loblolly pine (Pinus taeda L).were isolated at exponential growth stage.Influences of various concentrations of basal medium,levels of BA,and concentrations of inositol on the differentiation of embryonal suspensor mass (ESM),early stage somatic embryos (ESE) ,and lae stage somatic embryos (LSE) were investigated .A study of the effect of various concentrations of LP basal medium sowed that the optimal basal medium concentration of ESM,ESE,and LSE differentiation was 1.25 LP medium.The effects of various levels of BA and inositol showed that the optimal concentrations of BA for the formation of ESM,ESE and LSE were 4 mg/L ,2mg/L and 1mg/L,respectively ,and the optimal concentrations of inositol for the ESM ,ESE and LSM formation were 400mg/L,800mg/L and 1,200mg/L,respectively.

北亚热带次生栎林与火炬松人工林土壤微生物生物量碳的季节动态

在南京林业大学下蜀森林生态定位研究站次生栎林和火炬松人工林内,采用随机区组设计,连续两年测定了林下土壤微生物生物量碳、土壤温度、土壤湿度和林地凋落物输入量.结果表明:两种林分的土壤微生物生物量碳呈明显的季节性波动,均在植物生长旺季维持在较低水平,而在植物休眠季节维持在较高水平;在0～10cm土层内,火炬松林和次生栎林土壤微生物生物量碳变幅分别在267.8～459.8mg.kg-1和278.6～467.8mg.kg-1;土壤微生物生物量碳与土壤温度之间具有显著的负相关关系,而与森林地上凋落物输入量、土壤湿度无显著相关关系.说明该区域土壤微生物生物量碳的季节波动可能与土壤中有效碳及其它土壤养分的可利用状况、植物根系对营养的竞争以及林木的生长节律有关.

火炬松基因资源的育种值预测与选择

以引自美国的258个不同改良程度的火炬松家系作为试验材料,利用基于混合线性模型的BLUP方法对258个家系14年生树高和胸径的育种值进行预测。首先以不同的模型分别预测半同胞家系和全同胞家系的育种值,再通过构建亲缘关系矩阵预测所有个体的育种值,对各类群家系和个体育种值的差异进行分析。14年生时,在参试的各个类群中,入选率同为15%的情况下,各类群根据胸径育种值共选出优良家系39个;全同胞家系类群入选家系的胸径遗传增益最高,高于群体平均值1.40 cm,达8.56%。以配合选择方式选出优良单株77个,入选率为1%。入选的优良家系可从美国重复引种,为火炬松良种生产服务。根据火炬松核心-主群体育种系统的需要,入选的优良单株可用以充实第2代育种群体,其中10个进入核心育种群体,其余67个进入主群体。精确的BLUP预测值和基于个体育种值的选择将提高遗传改良进程中的选择效率和遗传增益,但在利用这些先进的统计分析方法提升林木良种选育效率的同时,必须对育种方案与试验设计进行改进完善。当参试家系数较多时,可采用平衡不完全区组设计,把所有参试家系统一在一个试验中,科学安排田间试验,以获得最大的成效。

不同抗性松树与松材线虫互作中H_2O_2及其氧化酶活性的差异

为探讨不同抗性松树接种松材线虫后过氧化氢(H2O2)、氧化酶活性变化差异与其抗性间的关系,对火炬松、马尾松和黑松接种松材线虫后针叶内H2O2含量、POD和APX酶活性动态变化进行了研究。结果发现,黑松和马尾松在接种早期(接种4 h后)针叶内H2O2含量(分别为(470.01±47.20)μmol/g和(232.33±9.42)μmol/g)远高于火炬松((145.46±11.86)μmol/g),但在接种72 h后,火炬松针叶内H2O2含量上升幅度较大,且在达到峰值后下降幅度比黑松和马尾松的小。3种松树接种松材线虫后的POD和APX酶活性均出现两次高峰值,峰值从大到小均为:黑松、马尾松、火炬松,且在达第1次活性高峰时感病松树上升快,在达第2次活性峰时抗病的火炬松上升快。这表明不同抗性松树接种松材线虫后,其体内H2O2含量、POD和APX酶活性变化差异与其抗性存在密切关系,H2O2含量的积累程度对松树抗病性具有重要的影响。

火炬松、湿地松、晚松组培繁殖的研究

本试验以火炬松、湿地松、晚松成熟种子的胚为材料，在附加有细胞分裂素（ＢＡＰ）的ＧＤ改良基本培养基（总盐分含量为０．２１％；总含氮量为１８６ｍｇ／Ｌ）经３～４周培养，在胚或子叶及下胚轴上出现膨大肉眼可见的暗绿色愈伤组织及小芽，其分化率达７５％～８５％。培养基中的细胞分裂素（ＢＡＰ）浓度对三种松种不定芽的分化具有很大影响，一般以３～５ｍｇ／Ｌ为好，在继代培养基中，活性炭对于嫩梢的生长具有良好的效果。当嫩梢长至１．５ｃｍ时，可供生根。在含有０．５ｍｇ／ＬＮＡＡ的生根培养基中经１５～２０ｄ培养，即长出白色愈伤组织，然后转至无激素培养基中培养１５～２０ｄ后生根。生根的小植株移栽入土壤，其成活率可达８０％～９５％。

火炬松胚性愈伤组织诱导和植株再生的研究

火炬松胚性愈伤组织诱导和植株再生的研究唐巍欧阳藩（中国科学院化工冶金研究所生化工程国家重点实验室北京１０００８０）郭仲琛（中国科学院植物研究所北京１０００９３关键词火炬松，体细胞胚胎发生，植株再生本文于１９９６年１０月２８日收到。国家“８６３”资...

火炬松核心育种群体子代生长变异与选择

广东省火炬松第1代核心育种群体68个全同胞组合的子代,组成了第2代核心群体的基本群体,在8,10,15年时分别对其树高、胸径和材积生长性状进行研究。结果表明:8,10,15年时,各生长性状在组合间都存在极显著差异。根据15年时各亲本子代的生长表现,选择8个优良母本和4个优良父本,可用于营建精选种子园或控制授粉种子园,生产遗传改良度高的种子。以15年全同胞家系平均材积作为选择指标,共选出10个组合,材积平均增益为18.94%,通过重复制种获得的种子可以用于营建插穗圃,通过扦插繁殖生产优良的全同胞家系苗木;采用配合选择共选出55个优良个体参与构建第2代育种群体,其中30个进入第2代核心群体,其余25个进入第2代主群体。

福建省火炬松人工林生长特性的研究

对福建省火炬松人工林生长特性以及不同栽培区火炬松的生长规律进行了对比分析。结果表明 :Richards函数是福建省火炬松优势木生长的最佳模型 ;对树高曲线方程而言 ,h=a0 da1 最符合火炬松树高随直径变化的规律。随着栽培区适宜程度的降低 ,火炬松胸径生长量随之降低 ,胸径平均生长量、材积连年生长量达到最大值的年龄也随之提早。

英德火炬松种子园子代在粤北的生长规律研究

Trials of 16-26 years old progenies from Yingde Loblolly Pine Seed Orchard were investigated by sample plot investigation.The trial fields were located at Yingde Loblolly Pine Seed Orchard,Yingde Forest Research Institute,Jiuqushui Forest Farm of Wengyuan County,Shaoguan Forest Farm and Renhua Forest Research Institute respectively.The growth process and performance level were studied by stem analysis based on average trees.Compared with that of local Masson pine(Pinus massoniana),the seedlots of loblolly pine(Pinus taeda) from Hubei and the seedlot of slash pine(P.elliottii) from Taishan Slash Pine Seed Orchard,the growth potential of progenies from Yingde Loblolly Pine Seed Orchard in North Guangdong was comprehensively evaluated.The results are as follows: The fast-growing period of height for the progenies from Yingde Loblolly Pine Seed Orchard began at the age of 3 and lasted for five to nine years.For diameter at breast-height(DBH) it began at the age of 5 and lasted for five to eight years.The fast-growing period for volume in Yingde Loblolly Pine Seed Orchard began at the age of 10 and lasted to 25 years old,while it began at the age of 10 both in Yingde Forest Research Institute and Jiuqushui Forest Farm,and at the age of 12 both in Shaoguan Forest Farm and Renhua Forest Research Institute.The progenies from Yingde Loblolly Pine Seed Orchard performed well generally in North Guangdong.Shaoguan and Renhua were suitable for the cultivation of large-dimension timber with the rotation longer than 25 years.In order to cultivate largedimension timber,suitable fertilizer shall be used when the DBH growth reaches the fast-growing period,and thinning operation shall be conducted when the mean annual increment for DBH reaches the maximum.

火炬松半同胞子代配合选择的遗传增益估算

以半同胞子代为材料,开展火炬松生长性状的评价并进行配合选择,在估算3种遗传力的基础上,估算配合选择的遗传增益。子代测定林分别建于广东英德(24°15′N,113°45′E)和浙江富阳(30°15′N,119°58′E)2个地点,包含24个共同家系,均来自广东英德火炬松种子园。子代林为随机区组设计,6次重复,英德试点为6株行式小区,富阳试点为8株行式小区。以SAS/STAT统计软件的VARCOMP过程用限制性最大似然法估算的方差成分进行遗传力的估算。结果表明:6年生树高单株遗传力、家系内遗传力和家系遗传力分别为0·161、0·129和0·579,胸径遗传力分别为0·075、0·059和0·384,单株材积遗传力分别为0·104、0·082和0·471。用3个公式估算配合选择的遗传增益。在数据严重不平衡的情况下,3个公式估算的结果非常接近,显示了简便的估算公式,即利用家系遗传力和家系内遗传力以及相应的选择差进行估算的公式,具有实用性和可靠性。

火炬松木材材性变异规律

研究分析了火炬松管胞形态和木材物理力学特征量的变异规律．得出了管胞长度、管胞直径、管胞长宽比、微纤丝角、生长轮宽度、晚材率、胞壁率和生长轮密度等材性指标变异规律的数学模型，为实现林木定向培育和速生优质制定科学集约经营措施以及材质的早期预测提供科学的理论依据。

火炬松树干液流的研究

采用ICT 2000TE树干液流测定系统,研究了密度为1020株/hm2的11年生火炬松秋季树干液流特征及其与环境因子的关系。结果表明:火炬松树干液流表现为单峰曲线,日出后,树干液流迅速上升,峰值出现在中午前后,日落后至次日日出前液流微弱或几近无上升液流;树干液流速度白天10时至17时平均约26cm/h,最大达到54cm/h;不同径阶火炬松树干液流流量差异极显著。4株样木的胸径分别为13.5,10.2,15.7,16.6cm,秋季耗水量分别为13.28,13.27,36.33,49.80kg/d。环境因子对树干液流的影响次序由大到小依次为:气温、相对湿度、光合有效辐射、土壤温度、土壤水分;逐步回归建立的方程达到极显著水平。

火炬松引种家系的遗传变异及速生稳定性综合评价

以国内早期引种的湿地松Pinuselliottii和福建乡土树种马尾松Pinusmassoniana为对照,对从美国种子园引种的21个火炬松Pinustaeda家系及国内早期引种种子园采种的4个火炬松家系进行遗传变异研究.结果表明:火炬松在福建省的总体生长表现优于对照湿地松和福建乡土树种马尾松,树高、胸径、材积等性状在家系间差异显著,且受中等以上遗传力所控制,遗传变异系数多在4 5%以上.火炬松生长性状5年生生长量的家系方差、地点方差和基因型与地点互作方差均达显著水平.通过对生产力指数和回归系数进行二维散点图聚类分析,最终确定25、19、18号家系为速生平均稳定家系,具有广泛的适应性,可在造林区普遍推广;20、17、23、24、22、16号家系为速生不稳定家系,特别适应于优良环境.

次生栎林和人工松林土壤呼吸对温度敏感性的室内模拟

土壤呼吸对温度的敏感性是影响全球气候变暖不确定性的因素之一。为了解在排除其他生态因子的影响下天然阔叶林和人工针叶林土壤呼吸对温度敏感性的差异，选择了苏南丘陵次生栎林和火炬松人工林为研究对象，采集2种林分0～10cm表层土壤，分别进行了5℃、15℃、25℃和35℃的室内恒温培养实验。用传统的密闭气室碱液吸收法测定它们的CO2释放量的动态变化，并测定土壤理化性质。结果表明：次生栎林对温度的敏感性即Q10值大于火炬松人工林；在5℃、15℃和25℃培养时2种林分土壤呼吸累积释放CO2量的差异不显著，并且土壤释放co：的动态变化一致，而35℃时，次生栎林显著大于火炬松人工林；在25℃培养时，次生栎林土壤呼吸累积释放co：量与5℃和15℃相比并没有显著增加，反而低于火炬松人工林；次生栎林的全碳、全氮、全钙、全磷含量以及碳氮比显著大于火炬松人工林。模拟结果表明，在全球气候变暖背景下，天然阔叶林可能比人工针叶林释放更多的CO2。