滨海沙地木麻黄优良无性系的引种推广试验

1994年从广东引进 3个木麻黄无性系 ,在本地育苗 ,小面积试验造林 ,按常规经营条件进行管理 ,各生长指标分别是本地品种 (对照 )的 2～ 7倍 ,大大超过部颁的木麻黄速生丰产林标准。引进的 3个木麻黄无性系比本地品种适应性好、生长快、抗性强 ,防风固沙效果好     

甘蔗S-腺苷甲硫氨酸合成酶基因(ScSAM)的克隆及表达

利用RT-PCR和RACE技术从甘蔗品种新台糖22号(ROC 22)中克隆获得SAM基因的cDNA序列,命名为ScSAM,GenBank登录号为KC172558。用生物信息学方法预测分析其序列,cDNA全长1466 bp,含有1个1191 bp的完整开放阅读框(ORF),编码396个氨基酸,与高粱和玉米等植物的SAM蛋白有很高的相似性。系统进化树分析显示,甘蔗ScSAM与高粱的SAM蛋白亲缘关系较近。Real-time PCR分析表明ScSAM为组成型表达,在根中的表达量最高,是叶中表达量的3.6倍。其在黑穗病菌胁迫和低温(4℃)、聚乙二醇(PEG)、NaCl非生物胁迫下均被诱导表达,但表达模式不同;在H₂O₂胁迫下其表达被抑制。推测其可能参与甘蔗抗黑穗病过程,且在甘蔗抗寒、抗旱、抗盐和抗氧化等胁迫过程中也起某种作用。     

半滑舌鳎HIRA基因部分cDNA序列克隆及表达分析

为研究HIRA基因在半滑舌鳎胚胎发育中的作用及其组织差异表达,以半滑舌鳎为研究对象,采用同源克隆策略从其精巢中分离了长度为764bp的HIRA部分cDNA序列,该片段编码254个氨基酸,具有5个wD结构域。对氨基酸进行比较发现,半滑舌鳎HIRA与比较物种的氨基酸序列具有很高的同源性,与红鳍东方纯亲缘关系最近。实时定量PCR分析发现,HIRA mRNA在多细胞期到原肠胚期的表达量较高,表明HIRA对胚胎发育起重要作用;HIRA在不同组织中表达差异显著,其中在性腺中表达最高,推测HIRA在维持性腺的功能方面有一定作用。     

小麦抗叶锈病Lr35基因同源序列RGA1侧翼序列的扩增与分析

利用小麦抗叶锈病近等基因系材料TcLr35,根据已扩增出的Lr35相关基因组DNA片段RGA1设计3对特异性引物,通过热不对称交错PCR技术获得2个有效扩增片段,将目的片段回收纯化,连接到pGEM-TEasy载体,转化EcoliDH5α感受态细胞中,经EcoRⅠ酶切,验证插入片段,选择合适菌液测序。获得了2条侧翼序列,长度分别为511bp和614bp,与RGA1重叠区分别为293bp和509bp,分别向3'端和5'端延长了208bp和100bp,拼接后序列为915bp。经BLASTn同源性比较与小麦抗叶锈病基因Lr21同源性为93%(score值为674,E值为0.0),与含有Lr10基因的450Kb大片段重叠群同源性为90%(score值为260,E值为7e-66),该研究为克隆Lr35目的基因奠定了基础。     

毛地黄优良变异植株组织培养及无性系的建立

研究了毛地黄优良变异植株嫩茎愈伤组织的诱导、分化及无性系的建立。结果表明:MS+1mg/L BA+0.6 mg/L 2,4-D是毛地黄嫩茎愈伤组织诱导的理想培养基;MS+0.5 mg/L BA+0.1 mg/LNAA是愈伤组织分化的理想培养基;MS+0.2 mg/L BA+0.2 mg/LIAA是不定芽壮苗培养的理想培养基;1/3 MS+0.3 mg/LIAA是生根培养的理想培养基;炉灰渣、河沙是生根试管苗移栽的理想基质。     

杉木无性系选育(开天系列)

在１５年间,从４￣６年生的半同胞和全同胞子代测验林及优良种源或种子园的子代林中选出近４００株优良个体,采用根颈萌条无性系化后建立选择性采穗圃,大量生产无性系苗开展多阶段的无性系比较试验。根据１０￣１２年（少数６年）生时的表现,从２５７个参试无性系中选出了１０个无性系。它们的单株材积比当地未去劣一代种子园或优良种源对照提高６０％￣１００％,而材质的平均表现基本与对照相当。这些无性系已生产规模推广,表     

耐盐基因HAL1的克隆及在原核细胞中的表达

以酵母基因组DNA为模板,采用PCR方法得到耐盐基因HAL1,插入原核表达载体pGEX-4T-1的XhoI和EcoRI酶切位点之间,构建原核表达载体pGEX-HAL1;将该载体转化到大肠杆菌中,重组菌株用IPTG诱导表达,其耐盐性比空白菌株提高50%;SDS-PAGE凝胶电泳结果显示有明显表达的蛋白质条带。     

Clone distribution of the earthworm Eiseniella tetraedra (Sav.) (Oligochaeta: Lumbricidae) across an altitudinal gradient on subarctic mountains of NW Europe

In earlier studies, we have shown that clone diversity of the parthenogenetic earthworm Eiseniella tetraedra increases from the upper reaches of rivers in northern Sweden towards their mouths. Now we survey brooks in the Scandes Mountains in the watershed between Sweden and Norway where major rivers originate. Using starch gel enzyme electrophoresis, we found 37 clones in a total catch of 379 individuals from six mountains. The most abundant clone made up 48.3% of the individuals collected. It was present on most mountains and was found at different elevations. In comparison with other clones it may represent a general-purpose genotype adapted to environmental conditions ranging from alpine through to subalpine to boreal habitats in the mountains. Diversity of clone assemblages decreased with increasing elevation. On four mountains, one to two clones were found at higher elevations. Passive downstream dispersal of E. tetraedra propagules from wider areas of the mountains was responsible for the more diverse clone pools in the lower reaches of the brooks (i.e., “small rivers behave like large ones”). Two clone groups, which deviated from the norm clone in their number of enzyme variants, were evenly distributed among different elevations. Therefore, we could not correlate genotype differences (i.e., adaptation of clones to mountain elevations). Clone pool similarities among the mountains were low on average (range 0–58%) but in a cluster of four mountains, similarities varied from 46% to 58%. Clone pool similarities between different elevations of the same mountain ranged from 27% to 83%. One mountain brook was sampled over 3 years to assess clone turnover. Only the norm clone was found in upstream habitats but it and three other clones were recorded downstream in at least 2 years. Ten clones were found once in the latter habitat as well.     

堇菜无性系建立的研究

为满足栽培对种苗的需求,以子叶为材料,采用组织培养的方法,对堇菜进行了愈伤组织诱导与分化、不定芽的分化、不定芽生根与试管苗生根继代增殖、试管苗的移栽和定植的研究,建立起堇菜的无性系。结果证明:MS+BA0.4mg/L+2,4-D丁酯2.5mg/L是子叶愈伤组织的诱导培养和继代增殖培养的理想培养基;MS+NAA0.1mg/L+BA0.6mg/L是愈伤组织分化培养和不定芽分化增殖继代培养的理想培养基;1/4MS+ABT2号2mg/L+NAA0.3mg/L是不定芽生根和试管苗生根继代增殖培养的理想培养基;试管苗移栽成活率为91.9%,定植成活率为99.1%。定植成活的试管苗生长旺盛,并保持了堇菜的所有植物学特征。     

绿竹咖啡酸-O-甲基转移酶基因(COMT)的克隆及相关分析

采用RT-PCR及RACE方法从绿竹中分离了COMT基因家族的一个基因,命名为BoCOMT1.BoCOMT1全长1 377 bp,编码一个361 aa的蛋白,估计分子量为39 kDa.BoCOMT1与小麦的TaCOMT、甘蔗的SoCOMT、玉米的ZmCOMT和毛白杨的PtCOMT的氨基酸序列比对结果表明,BoCOMT1与TaCOMT的氨基酸同源性最高,达到86.7%,系统进化树的分析表明,BoCOMT1与TaCOMT亲缘关系较近.RT-PCR结果显示,BoCOMT1在茎中的表达量约为叶中的2倍.